midwife-turned-immunization coordinator Nur Khalida has been based at Puskesmas Ingin Jaya on the outskirts of Banda Aceh City.   Nur works with local midwives on how best to improve immunization coverage focusing on addressing vaccine hesitancy.   \"I believe in using a personal approach to build trust with the community,” she explains “I always offer my phone number for follow-up questions which helps alleviate fears.”   Nur Khalida (left) and other participants engage in a role-play exercise These practical sessions on immunisation administration and role-playing exercises helped participants effectively explain diseases and vaccinations to parents recent outbreaks of vaccine-preventable diseases such as measles polio and diphtheria have highlighted the urgent need to improve immunization coverage just 20.8% of children in Aceh were fully immunized well below the national average of 95.3%.   well-trained and empowered health workers can boost vaccine acceptance protect communities and ultimately save lives.   “Health workers are the bridge between public health policies and the communities they serve,” explains Dr Muhammad Fathun WHO Technical Officer for vaccination in Aceh “It’s not enough to simply offer vaccines; they must be able to communicate educate and earn people’s trust.”  This simple yet crucial observation was the foundation of a Ministry of Health (MoH) and World Health Organization (WHO)-supported training in Banda Aceh City Trainers demonstrated how to identify frozen vaccines “Many people in the community fear adverse events following immunization or face opposition from family members particularly fathers,” says Dr Fathun we addressed these concerns with participants through practical role-playing exercises.”   frontline health workers are the ones who have the most direct contact with families so they need to be equipped with the right tools and communication skills to address community concerns in a compassionate and understanding way.\"  The MoH and WHO-supported training was conducted in two five-day batches bringing together 180 participants from 7 districts in the province This included 102 immunization officers and 78 midwives In addition to community engagement and education the training covered key skills such as microplanning and vaccine cold chain management including how to identify frozen vaccines.   Participants engage in hands-on record keeping and cold chain management training at a Puskesmas Sessions were also held on reporting immunization programme implementation surveillance of adverse events following immunization and monitoring and evaluating immunization activities Nur notes that both the content of the training and the participants marked a significant shift from earlier approaches.   training for central health office staff was prioritized while training for midwives and immunization officers was minimal.   the immunization officers often knowing who was vaccinated and who was not.    gaps are being bridged – one health worker at a time.    This activity was supported by the Australian government through the Department of Foreign Affairs and Trade "I believe in using a personal approach to build trust with the community,” she explains so they need to be equipped with the right tools and communication skills to address community concerns in a compassionate and understanding way."  Partly cloudy skies early will become overcast later during the night Want to get a weekly summary of New Jersey news from dozens of trustworthy news sites and not just one who has five decades in the news business and now publishes local site 70and73.com Outdoor dining venue at Ristorante Fieni's in Voorhees Ristorante Fieni’s has been a staple of the dining scene in South Jersey the popular Italian BYOB in Voorhees was immediately impacted by COVID-19 and its dining room's closure shifting to takeout and delivery was a no-brainer With outdoor dining now allowed and indoor dining shelved the restaurant has a positive outlook for now "Fortunately for us, we offered takeout before the pandemic and eventual shutdown. So we doubled down on what we offered and changed our whole system to accommodate carry out," Marco Fieni told 70and73.com "We spent the first few days stocking up on everything we possibly could and learning and tweaking as we went along." Recognizing that the shutdown would be long-term Fieni knew that adapting the way the restaurant did business and communicating with its customer base would be essential to keep the restaurant in business "We documented every step along the way across all our social media accounts," noted Fieni Getting the word out and keeping people updated was especially important when outdoor dining was initially permitted Fieni updates the restaurant’s Facebook page daily with photos of the outdoor patio dining area "We’ve offered outdoor seating on our patio for a few years now We always used it to help supplement demand especially on the weekends when the weather was nice," Fieni said "We have approximately 20 to 25 seats available outside depending on party sizes we can section off some areas of our parking lot as well." Striving to Thrive is a series of articles that look at how South Jersey businesses are faring in the COVID-19 pandemic and how they may change as we emerge from the crisis. Ideas or suggestions? Email contactus@70and73.com Outdoor dining has been a nice supplement to the takeout business the business has been solid overall without having to go back to a reservation-based system "We decided to accept walk-ins only because the weather is a factor," mentioned Fieni "The last thing we’d like to do is disappoint anyone." Eager to safely welcome back guests inside Fieni was ready to open up the dining room even at 25% capacity and strict guidelines on July 2 It would have provided a nice boost in business over the Fourth of July weekend but he’s optimistic this delay is not a catastrophe "It’s business as usual for outdoor dining and takeout with occasional requests for catering business." Safety is at the top of his mind and he’s prepared to keep doing what it takes to remain open and keeping staff and customers safe and healthy in the outdoor dining area "Tables are spaced six feet apart from one another and groups are capped at a maximum of eight guests per table," Fieni explained "All employees are required to wear masks and gloves in addition to hand washing and sanitizing We are using disposables as much as possible — menus peppers and other condiments in particular We also have sanitizer stations located throughout." Undeterred by all of the changes and continued uncertainty Fieni is hopeful the longtime family-owned and operated business will withstand these chaotic times "Things are changing and every day seems to be something new there are no recent results for popular images there are no recent results for popular videos Your browser is out of date and potentially vulnerable to security risks.We recommend switching to one of the following browsers: Account processing issue - the email address may already exist Invalid password or account does not exist Submitting this form below will send a message to your email with a link to change your password An email message containing instructions on how to reset your password has been sent to the email address listed on your account Metrics details The development of selective and nontoxic immunotherapy targeting prostate cancer (PC) is challenging Interleukin (IL)30 plays immunoinhibitory and oncogenic roles in PC and its tumor-specific suppression may have significant clinical implications CRISPR/Cas9-mediated IL30 gene deletion in PC xenografts using anti-PSCA antibody-driven lipid nanocomplexes (Cas9gRNA-hIL30-PSCA NxPs) revealed significant genome editing efficiency and circulation stability without off-target effects or organ toxicity Biweekly intravenous administration of Cas9gRNA-hIL30-PSCA NxPs to PC-bearing mice inhibited tumor growth and metastasis and improved survival Cas9gRNA-hIL30-PSCA NxPs suppressed ANGPTL 1/2/4 EGF and HGF expression in human PC cells while upregulated CDH1 leading to low proliferation and extensive ischemic necrosis IL30-targeting immunoliposomes downregulated NFKB1 expression and prevented intratumoral influx of CD11b+Gr-1+MDCs and prolonged host survival by inhibiting tumor progression This study serves as a proof of principle that immunoliposome-based targeted delivery of Cas9gRNA-IL30 represent a potentially safe and effective strategy for PC treatment given that PC mainly affects patients older than 50 years of age an increase in new cases is expected in the near future due to population aging great strides in therapy have not been made The development of personalized treatments that are well-tolerated by elderly patients with comorbidities is a major challenge and would have a high impact on the health care system many challenges remain to be addressed to increase its efficacy among which the delivery system plays a key role we designed and synthesized a nonimmunogenic biocompatible cationic lipid nanocomplex coated with PEG (NxP) and conjugated with anti-PSCA antibodies (Abs) for the selective delivery of the Cas9 guide (g) RNA-IL30 complex to human (h)-derived PSCA+IL30+ PC xenografts and in a syngeneic We demonstrated the efficient uptake of Ab-conjugated Cas9gRNA-IL30 NxPs in PC cells both in vitro and in vivo tumor-selective suppression of IL30 signaling pathways and remodeling of the intratumoral immune cell context resulting in substantial tumor growth inhibition and improved survival without evident toxicity Our investigation provides a proof of concept that immunoliposome delivery of the Cas9gRNA-hIL30 complex for IL30 genome editing at the tumor site is a clinically valuable tool for the safe and effective immunotherapy of PC The cell lines were cultured in RPMI-1640 with 10% FCS (Seromed confirmed to be mycoplasma-free by PCR analysis and passaged for fewer than 6 months after initial thawing PSCA expression on DU145, PC3, wild-type TRAMP-C1 and IL30-TRAMP-C1 cells and the conjugation efficiency between the anti-PSCA Abs and the aldehyde-modified DSPE-PEG2000 lipid present on the external layer of the NxP were assessed using flow cytometry, as described in the Supplementary Methods Cationic lipid nanocomplexes coated with PEG (NxP) were synthesized using a microfluidic device from Dolomite Microfluidics (Royston, UK) and lipids, as described in detail in the Supplementary Methods by the European Medicines Agency (EMA) and Food and Drug Administration (FDA) The encapsulation efficiency of Cas9gRNA-IL30 into nanoliposomes and the release rate of the Cas9IL30 NxPs were determined as reported in the Supplementary Methods Whole-genome next-generation sequencing (NGS) was performed by Lexogen GmbH (Wien Austria) using the Illumina NovaSeq 6000 platform system and Dragen software The intracellular uptake of NxPs by PC cells in vitro was analyzed using TEM, as reported in the Supplementary Methods PC cell viability and proliferation after treatment with empty NxPs or Cas9IL30-loaded NxPs were assessed using the CellTiter 96 AQueous One Solution Cell Proliferation Assay (#G3582; Promega USA) according to the manufacturer’s instructions Mice weight was constantly monitored for 63 days consistent with the longest treatment schedule duration (12 administrations Histopathological and ultrastructural analyses of the intracellular uptake and toxicity of NxPs in vivo were performed as described in the Supplementary Methods Laser scanning confocal (LSC) microscopy analyses of the lung transit and tumor uptake of the NxPs were performed as described in the Supplementary Methods 3 groups of 5 PC3 and 3 groups of 5 DU145 tumor-bearing NSG mice and 3 groups of 5 TRAMP-C1 tumor-bearing C57BL/6J mice were i.v treated when the tumors reached a mean volume of ~700 mm3 with Cas9hIL30 NxPs or Cas9mIL30 NxPs conjugated or not conjugated with anti-PSCA Abs or free Cas9 (in PBS) at a dose of 1.5 mg of Cas9 equivalent/kg blood was collected at predetermined time intervals (5 min The biodistribution of NxPs and their efficiency in targeting the tumor site were determined by using a 3T scanner (Philips Medical Systems, Nederland), as described in the Supplementary Methods Analysis of Cas9 protein in tumors and organs from NSG mice treated with anti-hPSCA Ab-conjugated or unconjugated NxPs was performed as reported in the Supplementary Methods Eight-week-old NSG male mice were purchased from Charles River (Wilmington, MA, USA) and housed under high barrier conditions according to the Jackson Laboratory’s guidelines (https://www.jax.org/jax-mice-and-services/find-and-order-jax-mice/nsg-portfolio/housing-and breeding-considerations-for-nsg-mice) in the animal facility of the Center for Advanced Studies and Technology at the “G To study the effects of IL30 target treatment on tumor growth and progression NSG male mice were subcutaneously (s.c.) injected with 5 × 105 wild-type (CTRL) DU145 cells or with 1 × 106 PC3 cells To study the effects of IL30-targeted treatment on the tumor immune microenvironment and tumor behavior three groups of twenty-seven 8-week-old C57BL/6J male mice were s.c injected with 5 × 105 mIL30 lentiviral-DNA-transfected TRAMP-C1 cells (IL30-TRAMP-C1) Mice were treated with Cas9IL30-PSCA NxPs, empty-PSCA NxPs, or PBS as soon as the tumors became palpable and twice a week thereafter, as shown in Supplementary Fig. 3 Two additional groups of twenty-seven 8-week-old NSG male mice were s.c injected with nontargeting guide RNA-transfected (NTgRNA) or IL30 knockout (IL30KO) DU145 or PC3 cells whereas an additional group of twenty-seven 8-week-old C57BL/6J male mice was s.c These control groups were monitored for tumor growth but were left untreated Tumors were measured with calipers as soon as they were palpable (2 mm in diameter) Based on the tumor growth and progression rates 12 mice from each group were euthanized at (4) key time points (3 mice per point) for histopathological and ultrastructural analyses The remaining 15 mice per group were kept alive until tumors reached 2 cm3 or evidence of suffering was observed Autopsy and histopathological examinations of the different organs (heart An overall sample size of 15 mice per group allowed the detection of a statistically significant difference in tumor growth among the five groups (ANOVA) with 80% power at a 0.05 significance level (G*Power The animal procedures were performed in accordance with the European Community and ARRIVE guidelines and were approved by the Institutional Animal Care Committee of “G d’Annunzio” University and by the Italian Ministry of Health (Authorization n between-group differences were assessed using the Student’s t test or ANOVA Survival curves were constructed using the Kaplan‒Meier method and survival differences were analyzed using the log-rank test All the statistical tests were evaluated at an α level of 0.05 using Stata and brain on both the T1- and T2-weighted images were calculated at scheduled time points for each group The results are expressed as the mean ± standard deviation CA) was used to perform all the statistical analyses All animal procedures were performed in accordance with the European Community and ARRIVE guidelines and were approved by the Institutional Animal Care Committee of “G The human prostate tissue samples for immunostaining with anti-PSCA Abs were obtained from the institutional Biobank of the Local Health Authority n The personal data processing complied with the data protection laws c Specific binding of anti-hPSCA Ab-conjugated/rhodamine B-labeled nanoliposomes (RhB-hPSCA-NxP) to the surface of DU145 (top right picture) and PC3 (bottom right picture) cells compared to unconjugated/rhodamine B-labeled nanoliposomes (RhB-NxP) (top left and bottom left pictures) Blue areas: anti-hPSCA Ab-conjugated or unconjugated NxPs The experiments were performed in triplicate d The immunoliposome used in this study consists of a bilayer phospholipid spheroid vesicle containing the CRISPR/Cas9gRNA-hIL30 complex and functionalized with anti-hPSCA Abs linked to PEGylated lipids f DLS analysis of the zeta potential of empty-hPSCA NxP (29.17 ± 3.56 mV) (e) and of Cas9hIL30-hPSCA NxP (3.42 ± 1.49 mV) (f) g Electron microscopy analysis of nanoparticles cultured with DU145 cells showing their spherical shape and submicron size a magnified detail of the image illustrating the ultrastructural features of the NxPs The values of the scale bars are reported in the images h–j Ultrastructural features of DU145 cells untreated (h) or treated with Cas9hIL30-hPSCA NxPs (i) which contain spherical-shaped vesicles composed of one or more phospholipid bilayers (arrowheads) j The image shows a magnification of one of these vesicles located close to the nuclear envelope k Ultrastructural features of DU145 cells that were not treated (a Similar to that observed in the untreated PC cells (a the treated cells contained intact mitochondria and endoplasmic reticulum (c–f) with no signs of ultrastructural damage to the cytoplasmic organelles the unconjugated NxPs largely remained on the tumor cell surface (c whereas the anti-hPSCA Ab-conjugated NxPs were endocytosed by the tumor cell and were mostly found in the cell cytoplasm (e spherical vesicles containing more phospholipid bilayers (arrows) were detected in the cytoplasm of both tumor cells treated with Ab-conjugated (f) and unconjugated (d) NxPs These results are comparable to those obtained with PC3 cells NxPs were loaded with the Cas9gRNA-hIL30 complex which demonstrated a fair entrapment efficiency (58%) when considering the substantial size of the Cas9 protein (160 kDa) and then conjugated with anti-hPSCA Abs to generate Cas9gRNA-hIL30-hPSCA NxPs hereinafter referred to as Cas9hIL30-hPSCA NxPs NxPs were left empty and conjugated with anti-PSCA Abs to generate empty-hPSCA NxPs both the Ab-conjugated and unconjugated NxPs had been endocytosed by the tumor cells a On-target and off-target characterization of CRISPR/Cas9gRNA-mediated hIL30 editing delivered using immunoliposomes in vitro Average frequency of CRISPR/Cas9-induced variants in the IL30 gene (editing efficiency or on-target effects OnT) and in off-target sites corresponding to recognized genetic loci (off-target effects OTs) in DU145 and PC3 cell cultures treated with Cas9hIL30-hPSCA NxPs The frequency of variants (OnT and OTs) in cells treated with PBS or Empty-hPSCA NxPs (controls) was <0.1% c On-target and off-target characterization of CRISPR/Cas9gRNA-mediated hIL30 editing by immunoliposomes in vivo OnT) and in off-target sites (off-target effects OTs) in the indicated organs of DU145 (b) and PC3 (c) tumor-bearing NSG mice treated with Cas9hIL30-hPSCA NxPs The frequency of variants (OnT and OTs) in the organs of mice treated with PBS or empty-hPSCA NxPs (controls) was <0.1% e Serum (d) and pH (e) stability of Cas9hIL30-hPSCA NxPs measured at different time points over a 24-h period f Cas9 release profile of Cas9gRNA-hIL30 NxP g Viability of DU145 (green bars) and PC3 (light blue bars) cells after 48 hrs of incubation with different concentrations (0.2 or 1.0 mg/ml) of empty-hPSCA NxPs versus PBS-treated cells The results obtained from untreated cells were comparable to those from PBS-treated cells h Pharmacokinetics of free Cas9 and Cas9hIL30 NxPs %ID/g = percentage of total injection dose per weight i Quantitative analysis of T1-weighted MR images of tumors from mice treated with CE-Gd-hPSCA NxPs versus tumors from mice treated with CE-Gd NxPs at different time points after nanoparticle injection Tukey HSD test versus tumors of mice injected with unconjugated NxPs at the same time point j In vivo T1-weighted MR images of tumor-bearing NSG mice before (a) and 2 hrs 30 min after intravenous injection with Gd-DOTA (b) The tumors are indicated with red dotted circles MRI data obtained from DU145 tumor-bearing mice are comparable to those obtained from PC3 tumor-bearing mice All experiments were performed in triplicate k Quantitative analysis of T1-weighted MR images of tumors versus organs in mice treated with CE-Gd NxPs at different time points after nanoparticle injection Signal intensity is reported as a percentage of the injected dose (%ID) Tukey HSD test versus organs at the same time point Tukey HSD test versus tumor and other organs at the same time point l Quantitative analysis of T1-weighted MR images of tumors versus organs in mice treated with CE-Gd-hPSCA NxPs at different time points after nanoparticle injection Tukey HSD test versus other organs at the same time point n Confocal microscopy images of lungs labeled with anti-EpCAM Abs (green) from DU145 tumor-bearing NSG mice Similar results were obtained in mice inoculated with RhB-NxPs which recommends a release rate of less than 40% at 0.5 hr Toxicity screening of the immunoliposomes assessed in both DU145 and PC3 cells exposed to increasing concentrations (0.2, 0.4, or 1 mg/ml) of empty-hPSCA NxPs for 48 hrs, revealed that the viability of PC cells treated with NxPs and PBS (or untreated) was comparable (ANOVA: p > 0.05), indicating that the immunoliposome formulation had no obvious toxicity (Fig. 2g) a–c Confocal microscopy images of tumors from PBS-treated (a) or RhB-labeled nanoparticle-treated (b c) mice showing RhB-hPSCA-NxP (red) or RhB-NxP (red) uptake by GFP-labeled DU145 tumors (green) Images of tumors from PBS-treated mice were comparable at each time point Similar results were obtained for GFP-labeled PC3 tumor-bearing NSG mice inoculated with RhB-hPSCA-NxPs or RhB-NxPs e Quantification of NxP uptake in DU145 (d blue bars) tumors that developed in NSG mice using LSC microscopy Tumor uptake of the RhB-NxPs (light green or blue) and RhB-hPSCA-NxPs (dark green or blue) is expressed as the mean percentage ± SD of RhB+GFP+ cells/total number of GFP+ cells Student’s t test vs RhB-NxPs at the same time point g TEM images of tumors 2 hrs 30 min after treatment of mice with Cas9hIL30-hPSCA NxPs (f) or Cas9hIL30 NxPs (g) showing that unconjugated NxPs (arrows) were poorly endocytosed and predominantly localized among tumor cells the majority of anti-hPSCA Ab-conjugated NxPs were endocytosed and primarily found in the cell cytoplasm (arrows) the effects of anti-hPSCA Ab-conjugated Cas9hIL30 NxPs were compared to those of anti-hPSCA Ab-conjugated empty NxPs to assess the antitumor potential of targeted delivery of the Cas9gRNA-hIL30 complex The treatment schedule consisted of two i.v. injections of immunoliposomes per week (250 µl dose with a 10 mg/ml lipid concentration) starting from the onset of a palpable tumor (~2 mm Ø) and continuing throughout the lifespan of the mice (Fig. 4a). a Treatment schedules for NSG mice bearing human-derived subcutaneous PC3 (blue arrow) or DU145 (green arrow) tumors that express membrane-anchored IL30 treatment with two weekly doses of NxPs (10 mg/ml) started when the tumors were palpable (Ø 2 mm) and stopped when the tumors reached 2 cm3 and the mice were sacrificed b The mean volume of subcutaneous tumors developed in 4 groups of fifteen NSG male mice after s.c implantation of IL30KO or wild-type PC3 cells and treatment with PBS Tukey HSD test versus PC3 tumors treated with PBS or empty-hPSCA NxPs The results obtained from tumors treated with Cas9-NTgRNA-hPSCA NxPs were comparable to those from tumors treated with empty-hPSCA NxPs The results are expressed as the mean ± SD c The mean volume of subcutaneous tumors developed in 4 groups of 15 NSG male mice after s.c implantation of IL30KO or wild-type DU145 cells and treatment with PBS Tukey HSD test versus DU145 tumors treated with PBS or empty-hPSCA NxPs or Cas9hIL30-hPSCA NxPs Tukey HSD test versus DU145 tumors treated with PBS or with empty-hPSCA NxPs d Kaplan–Meier survival curves of 4 groups of fifteen NSG male mice bearing tumors developed after s.c implantation of IL30KO PC3 cells or wild-type PC3 cells and treated with empty-hPSCA NxPs and mice bearing IL30KO-PC3 tumors survived longer than mice bearing wild-type PC3 tumors treated with PBS or empty-hPSCA NxPs (63 versus 47 days; log-rank test: p < 0.000001) Mice were sacrificed when tumors reached 2 cm3 in size e Kaplan–Meier survival curves of 4 groups of fifteen NSG male mice bearing tumors developed after s.c implantation of IL30KO DU145 cells or wild-type DU145 cells and treated with empty-hPSCA NxPs Mice bearing wild-type DU145 tumors treated with Cas9hIL30-hPSCA NxPs survived longer than mice bearing wild-type DU145 tumors treated with PBS or empty-hPSCA NxPs (74 versus 58 days) but less than mice bearing IL30KO-DU145 tumors (94 days) (log-rank test: p < 0.000001) The mice were sacrificed when the tumors reached 2 cm3 in size f Percentages of mice which developed lung metastases (>500 µm) or micrometastases (≤500 µm) in wild type (or IL30KO) PC3 tumor-bearing NSG mice treated with Cas9hIL30-hPSCA NxPs or Empty-hPSCA NxPs The results from PBS-treated wild-type PC3 tumor-bearing mice were comparable to those from empty-NxP-treated mice p = 0.0000000012 versus mice bearing wild-type PC3 tumors and treated with PBS g Histopathological features of lung metastasis (M) that developed in empty-hPSCA NxP-treated mice bearing PC3 tumors (a) and lung micrometastases (m) that developed in PBS-treated mice bearing IL30KO-PC3 tumors (b) h Percentage of lung metastases spontaneously developed in wild-type (or IL30KO) DU145 tumor-bearing NSG mice treated with Cas9hIL30-hPSCA NxPs or empty-hPSCA NxPs The results from PBS-treated wild-type DU145 tumor-bearing mice were comparable to those from empty-hPSCA NxP-treated mice p = 0.0003 versus mice bearing wild-type DU145 tumors and treated with PBS or Empty-hPSCA NxPs i Histopathological features of lung metastasis (M) in empty-hPSCA NxP-treated mice bearing DU145 tumors (a) and healthy lung tissue from Cas9hIL30-hPSCA NxP-treated mice bearing DU145 tumors To compare the effects on tumor growth and progression of the immunoliposome-based delivery of CRISPR/Cas9gRNA-hIL30 complex versus a direct IL30 gene editing in PC cells which generated IL30KO-DU145 and IL30KO-PC3 cells in addition to three groups of NSG (NOD scid gamma) mice s.c implanted with wild-type DU145 or PC3 cells and then treated with Cas9hIL30-hPSCA NxPs or PBS; two groups of mice implanted with IL30KO-DU145 or IL30KO-PC3 cells b Subcutaneous PC3 (a) and DU145 (b) tumors from Cas9hIL30-hPSCA NxP-treated animals show the absence of IL30 expression (a and b multiple areas of vascular leakage (see the inset in a b) and ischemic-coagulative necrosis (a and b b) associated with defective vascularization (a and b c) and low cancer cell proliferation (a and b as observed in IL30KO tumors from PBS-treated mice (a tumors from Empty-hPSCA NxP-treated mice expressed IL30 (a and exhibited a well-developed vascular network (a Apoptotic events were comparable in tumors from the different treatment groups (a The results obtained from wild-type PC3 or DU145 tumors developed in PBS-treated mice were comparable to those of tumors from empty-PSCA-treated mice c DU145 (green bars) and PC3 (light blue bars) cell viability after 72 h of incubation with (1.0 mg/ml) Cas9hIL30-hPSCA NxPs versus untreated and IL30KO cells Tukey HSD test versus Cas9hIL30-hPSCA NxP-treated cells and untreated cells The results from untreated cells were comparable to those obtained from Empty-hPSCA NxP-treated cells Fold differences in the mRNAs of angiogenesis-related genes between IL30KO-DU145 cells and control NTgRNA-treated DU145 cells (d; light green bars) or PC3 cells (e; light blue bars) and between Cas9hIL30-hPSCA NxP-treated DU145 cells and control empty-hPSCA NxP-treated DU145 cells (d; dark green bars) or PC3 cells (e; dark blue bars) A significant threshold of a twofold change in gene expression corresponded to p < 0.001 Only genes with a fold change >2 are shown The experiments were performed in duplicate The dashed lines represent the twofold change cutoff f Venn diagram representing the “Angiogenesis Driver Genes” which are downregulated (red arrows pointing downward) or upregulated (blue arrows pointing upward) by IL30KO in DU145 (green circle) and PC3 cells (blue circle) and by Cas9hIL30-hPSCA NxP treatment of DU145 (purple circle) and PC3 cells (red circle) Overlapping circles illustrate the shared IL30-regulated genes between the different IL30 gene-editing approaches in the different cell lines Immunopathological detection of the downregulation of angiogenic drivers in IL30KO tumors and nanoparticle-treated tumors substantiated the molecular in vitro findings b Expression of the proangiogenic genes IGF1 (g c and d) was minimal to absent in Cas9hIL30-hPSCA NxP-treated PC3 (g a and d) and in Cas9hIL30-hPSCA NxP-treated DU145 (h a and c) tumors compared to empty-hPSCA NxP-treated PC3 (g b and e) and empty-hPSCA NxP-treated DU145 (h The immunohistochemical features of Cas9hIL30-hPSCA NxP-treated PC3 tumors (g a and d) were similar to those observed in PBS-treated IL30KO-PC3 tumors (g The immunohistochemical features of the control tumors that developed in the NSG after implantation of the NTgRNA-treated cells were comparable to those of the empty-hPSCA NxP-treated tumors and to those of the untreated wild-type tumors d Subcutaneous PC3 (a) and DU145 (b) tumors from Cas9hIL30-hPSCA NxP-treated animals were characterized by marked inhibition of proangiogenic genes were strongly expressed compared to empty-hPSCA NxP-treated tumors (a: b The immunopathological features of Cas9hIL30-hPSCA NxP-treated PC3 tumors (a: a s) were similar to those of PBS-treated IL30KO-PC3 tumors (a: c The immunopathological features of control tumors that developed in NSGs after implantation of NTgRNA-treated cells were comparable to those of empty-hPSCA NxP-treated tumors and untreated wild-type tumors b TEM analyses of subcutaneous tumor xenografts and kidneys from PC3 (a) and DU145 (b) tumor-bearing NSG mice treated with Cas9hIL30-hPSCA NxPs Compared with the tumors of the PBS-treated mice (a the tumors of the empty-hPSCA NxP- or Cas9hIL30-hPSCA NxP-treated mice exhibited cytoplasmic vacuoles that were consistent with aspects of nanoparticle endocytosis (arrows) and renal tubules of treated mice showed no signs of cell damage or ultrastructural features comparable to those of the respective organs of PBS-treated mice and prostate (d) of DU145 tumor-bearing NSG mice treated with Cas9hIL30-hPSCA NxPs (a and similar to those observed in empty-hPSCA NxP-treated mice (b Similar results were obtained from the histopathology of the organs of PBS-treated mice Cas9hIL30-hPSCA NxPs had no obvious hemolytic effect the immunoliposome-based targeted delivery of the Cas9gRNA-hIL30 complex which selectively inhibits tumor IL30 expression has antiangiogenic and antiproliferative effects in both PC models and has antimetastatic efficacy in DU145 tumor model without evidence of hematological or organ toxicity These cells produced rapidly progressing and highly metastatic tumors resulting in decreased survival compared to that of mice bearing control tumors (wild-type TRAMP-C1 and EV-TRAMP-C1) a PSCA expression in tumors developed after s.c implantation of wild-type TRAMP-C1 and IL30-TRAMP-C1 cells in syngeneic C57BL/6J mice b Specific binding of anti-mPSCA-conjugated/rhodamine B-labeled nanoliposomes (RhB-mPSCA-NxPs) to the surface of IL30-TRAMP-C1 (right image) cells compared to unconjugated/rhodamine B-labeled nanoliposomes (RhB-NxPs) (left image) Blue areas: anti-mPSCA-conjugated or -unconjugated NxPs c Ultrastructural features of IL30-TRAMP-C1 cells untreated (a) or treated with Cas9mIL30 NxPs (b The nanoparticles appear spherical and have a regular the unconjugated NxPs largely remained on the tumor cell surface (b showing poor penetration into the cytoplasm (c the anti-mPSCA-conjugated NxPs were endocytosed by the tumor cell and were mostly found in the cell cytoplasm (d d On-target and off-target characterization of CRISPR/Cas9gRNA-mediated Il30 editing in vitro Average frequency of CRISPR/Cas9gRNA-induced variants in the Il30 gene (editing efficiency or on-target effects OTs) in IL30-TRAMP-C1 cell cultures treated with Cas9mIL30-PSCA NxPs or Cas9mIL30-NxPs (total editing efficiency of 82% and 12% The frequency of variants (OnT and OTs) in cells treated with empty-mPSCA NxPs or PBS (controls) was <0.1% e On-target and off-target characterization of CRISPR/Cas9gRNA-mediated Il30 editing in vivo OnT) and in off-target sites (OTs) in the indicated organs of IL30-TRAMP-C1 tumor-bearing C57BL/6J mice treated with Cas9mIL30-mPSCA NxPs The frequency of variants (OnT and OTs) in the organs of mice treated with empty-mPSCA NxPs or PBS (controls) was <0.1% f Sequencing of the sgRNA-targeted Il30 locus and mutation spectrum analysis using ICE The image shows the percentage of frameshift or 21+ bp indels identified in IL30-TRAMP-C1 cells treated with Cas9mIL30-mPSCA NxPs or Empty-mPSCA NxPs The data obtained from PBS-treated cells were comparable to those obtained from cells treated with empty-mPSCA NxPs g Pharmacokinetics of free Cas9 and Cas9mIL30 NxPs conjugated or not conjugated with anti-mPSCA Abs in IL30-TRAMP-C1 tumor-bearing C57BL/6 J mice h TEM analyses of syngeneic tumors from C57BL/6J mice treated with Cas9mIL30-mPSCA NxPs revealed spherical vesicles that were consistent with those of nanoparticles and close to (arrows) or inside (arrowheads) the tumor cells i The mean volume of wild-type and IL30-TRAMP-C1 subcutaneous tumors developed in C57BL/6J mice after treatment with Cas9mIL30-mPSCA Tukey HSD test versus IL30-TRAMP-C1 tumors treated with PBS or empty-mPSCA NxPs and PBS-treated wild-type TRAMP-C1 tumors Tukey HSD test versus IL30-TRAMP-C1 tumors treated with PBS or Empty-mPSCA NxPs j Incidence of lung metastases in C57BL/6J mice bearing subcutaneous IL30-TRAMP-C1 tumors treated with Cas9mIL30-mPSCA p = 0.0005 versus IL30-TRAMP-C1 + empty-mPSCA NxPs k Kaplan–Meier survival curves of 4 groups of fifteen C57BL/6J male mice bearing tumors developed after subcutaneous implantation of IL30-TRAMP-C1 cells or wild-type TRAMP-C1 cells treated with empty-mPSCA NxPs Mice bearing IL30-TRAMP-C1 tumors treated with Cas9mIL30-mPSCA NxPs survived longer (92 days) than mice bearing IL30-TRAMP-C1 tumors treated with PBS or empty-mPSCA NxPs (64 days) or mice bearing wild-type TRAMP-C1 tumors treated with PBS (78 days) (log-rank test: p < 0.000001) l TEM images of kidneys (a) and liver (b) from IL30-TRAMP-C1 tumor-bearing C57BL/6J mice treated with Cas9mIL30-mPSCA NxPs showing no signs of ultrastructural cell damage m Subcutaneously developed IL30-TRAMP-C1 tumors from Empty-mPSCA NxP-treated animals (top panel) show a solid growth pattern with no signs of vascular damage (a) rich vascularization (b) and strong IL30 expression (c) tumors from Cas9mIL30-mPSCA NxP-treated animals (bottom panel) showed multiple areas of vascular leakage (d which were associated with defective vascularization (e) and low to absent IL30 expression (f) n Subcutaneously developed IL30-TRAMP-C1 tumors from Empty-mPSCA NxP-treated animals (top panel) showed considerable Foxp3+Treg lymphocyte (a) and NKp46+RORγt+ ILC3 (b) infiltration and high cytoplasmic and nuclear NFKB1 expression (c) Tumors from Cas9mIL30-mPSCA NxP-treated animals (bottom panel) showed few Foxp3+ Treg lymphocytes (d) and NKp46+RORγt+ ILC3 (e) and lower cancer cell expression of NFKB1 (f) compared to tumors from empty-mPSCA NxP-treated animals (c) The serum and pH stabilities of the Cas9mIL30-mPSCA NxPs were comparable to those of the Cas9hIL30-hPSCA NxPs and the release profile of Cas9 by Cas9mIL30 NxPs was comparable to that of Cas9hIL30 NxPs The effect of Cas9mIL30-mPSCA NxPs on tumor production of IL30 as well as on the tumor microenvironment and biological behavior This experiment included one group of mice implanted with wild-type TRAMP-C1 cells and then treated with PBS in addition to three groups of C57BL/6J mice implanted with IL30-TRAMP-C1 cells and then treated with Cas9mIL30-mPSCA NxPs Assessment of NxP immunogenicity in fully immunocompetent C57BL/6J mice demonstrated that i.v inoculation of naked Cas9gRNA-mIL30 complex or Cas9mIL30-mPSCA NxPs induced a minimal increase in the blood levels of TNFα and IL6 Immunotherapy has been a breakthrough in the treatment of advanced tumors29 but limited results have been obtained in PC The identification of new targetable immunosuppressive oncogenes would be useful Given that PC mainly affects the fragile and progressively expanding elderly population a tumor-selective and well-tolerated therapeutic approach is especially needed Nanobiotechnology can provide innovative tools for the development of precision cancer immunotherapy and IL30 represents a potential therapeutic target in PC ensures efficient site-specific accumulation and favors internalization into PC cells excluded the possibility of organ damage or obvious inflammatory responses These data were substantiated by the lack of significant changes in weight and serum levels of inflammatory cytokines and markers of cardiac Chronic biweekly treatment with immunoliposomes substantially inhibited tumor growth and progression After traveling through the pulmonary circulation intravenously administered immunoliposomes enter the systemic circulation but exclusively accumulate in PSCA+ tumors yielding inhibitory effects on tumor progression programs Remodeling the TME in immunocompetent hosts is associated with substantial inhibition of tumor growth and metastasis leading to improved survival of IL30-targeting immunoliposome-treated mice and strengthening the results obtained in xenograft tumor models immunoliposomes that selectively target tumors and deliver a CRISPR/Cas9 complex to edit tumor-promoting and immunoregulatory genes are absent from the current therapeutic landscape emphasizing the clinical value of an IL30-targeting therapeutic strategy The dramatic disruption of proangiogenic and tumor progression programs and the overcoming the immunosuppressive TME attributed to the selective nanoliposome-mediated deletion of the IL30 gene in cancer cells paves the way for the development of a modern and safe immuno-nanotherapy for advanced PC that meets the needs of healthy aging The data generated in this study are available upon request from the corresponding author Nanomaterials for cancer therapy: current progress and perspectives A review of liposomes as a drug delivery system: current status of approved products Peptide-functionalized liposomes as therapeutic and diagnostic tools for cancer treatment Novel liposomal formulation of baicalein for the treatment of pancreatic ductal adenocarcinoma: design a heterodimeric cytokine composed of EBI3 and p28 protein induces proliferation of naive CD4+ T cells Interleukin-30 expression in prostate cancer and its draining lymph nodes correlates with advanced grade and stage Interleukin-30 promotes breast cancer growth and progression Interleukin-30/IL27p28 shapes prostate cancer stem-like cell behavior and is critical for tumor onset and metastasization A folding switch regulates interleukin 27 biogenesis and secretion of its α-subunit as a cytokine Interleukin-30 feeds breast cancer stem cells via CXCL10 and IL23 autocrine loops and shapes immune contexture and host outcome CRISPR/Cas9-mediated deletion of Interleukin-30 suppresses IGF1 and CXCL5 and boosts SOCS3 reducing prostate cancer growth and mortality CRISPR-Cas guides the future of genetic engineering CRISPR-Cas9: a multifaceted therapeutic strategy for cancer treatment Isolation of a human prostate carcinoma cell line (DU 145) PC3 is a cell line characteristic of prostatic small cell carcinoma Characterization of prostatic epithelial cell lines derived from transgenic adenocarcinoma of the mouse prostate (TRAMP) model Rapid quantitative evaluation of CRISPR genome editing by TIDE and TIDER Inference of CRISPR edits from Sanger trace data Prostate stem cell antigen (PSCA) expression increases with high Gleason score advanced stage and bone metastasis in prostate cancer Prostate stem cell antigen is overexpressed in prostate cancer metastases In Nanomaterials Safety: Toxicity And Health Hazards (ed Intravital microscopy through an abdominal imaging window reveals a pre-micrometastasis stage during liver metastasis Recent insights into NF-κB signalling pathways and the link between inflammation and prostate cancer Complete loss of PTEN expression as a possible early prognostic marker for prostate cancer metastasis 10% Body weight (gain) change as criterion for the maximum tolerated dose: a critical analysis Hindering triple negative breast cancer progression by targeting endogenous interleukin-30 requires IFNγ signaling A guide to cancer immunotherapy: from T cell basic science to clinical practice Targeting Interleukin(IL)-30/IL-27p28 signaling in cancer stem-like cells and host environment synergistically inhibits prostate cancer growth and improves survival The new frontier of genome engineering with CRISPR-Cas9 Current advances toward the encapsulation of Cas9 Stealth liposomes: review of the basic science Advanced strategies in liposomal cancer therapy: problems and prospects of active and tumor specific drug release PEGylation as a strategy for improving nanoparticle-based drug and gene delivery The role of the helper lipid on the DNA transfection efficiency of lipopolyplex formulations Lipid nanoparticles functionalized with antibodies for anticancer drug therapy Interleukin-30 subverts prostate cancer-endothelium crosstalk by fostering angiogenesis and activating immunoregulatory and oncogenic signaling pathways Insulin-like growth factor 1 (IGF-1) stabilizes nascent blood vessels CCN2/connective tissue growth factor is essential for pericyte adhesion and endothelial basement membrane formation during angiogenesis CXC chemokines in cancer angiogenesis and metastases Emerging role of endothelin-1 in tumor angiogenesis Inhibition of angiogenesis and tumor progression of MK-0429 TNF primes endothelial cells for angiogenic sprouting by inducing a tip cell phenotype Role of matrix metalloproteinases in angiogenesis and cancer Fibroblast growth factor regulation of neovascularization Cyclooxygenase-2: a therapeutic target in angiogenesis Tumor suppressor gene E-cadherin and its role in normal and malignant cells CRISPR-mediated reactivation of DKK3 expression attenuates TGF-β Signaling in prostate cancer Dickkopf-3 in human malignant tumours: a clinical viewpoint PTEN: tumor suppressor and metabolic regulator The role of regulatory T cells in the pathogenesis and treatment of prostate cancer RORγt+ innate lymphoid cells promote lymph node metastasis of breast cancers IL23-producing human lung cancer cells promote tumor growth via conversion of innate lymphoid cell 1 (ILC1) into ILC3 Plasticity of innate lymphoid cell subsets Modes of resistance to anti-angiogenic therapy Resistance mechanisms to anti-angiogenic therapies in cancer Combination of anti-angiogenic therapy and immune checkpoint blockade normalizes vascular-immune crosstalk to potentiate cancer immunity Determinants of resistance to VEGF-TKI and immune checkpoint inhibitors in metastatic renal cell carcinoma Nanomedicine review: clinical developments in liposomal applications Liposomal nanomedicine: applications for drug delivery in cancer therapy Immunotherapy for prostate cancer: a current systematic review and patient centric perspectives Molecular targeted therapies in metastatic prostate cancer: recent advances and future challenges Download references The research leading to these results has received funding from the following: AIRC Emma Di Carlo; MUR (Ministero dell’Università e della Ricerca) from the National Operational Program for Research and Innovation/Programma Operativo Nazionale Ricerca e Innovazione 2014–2020 (PON R&I) to Carlo Sorrentino; the European Union - Next Generation EU under the National Recovery and Resilience Plan (NRRP) digitalisation and sustainability for the diffused economy in central Italy” to Antonella Fontana Samanta Moffa for helping in the preparation of vesicles and their characterization Department of Medicine and Sciences of Aging Anatomic Pathology and Immuno-Oncology Unit Center for Advanced Studies and Technology (CAST) Institute for Advanced Biomedical Technologies (ITAB) wrote the manuscript and was responsible for the overall content The authors declare no competing interests Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations Download citation DOI: https://doi.org/10.1038/s12276-024-01310-2 Anyone you share the following link with will be able to read this content: a shareable link is not currently available for this article Prostate cancer is an age-related disease and a leading cause of cancer death in men worldwide Although its incidence has remained stable over the past decade the proportion of tumors diagnosed in the advanced stages is rising highlighting the need for improved and personalized treatments especially for older patients with additional health problems Di Carlo and coworkers created a NanoLiposome delivery system to transport a gene-editing tool to target and inhibit a particular gene involved in cancer growth This study tested this approach’s effectiveness and safety on human prostate cancer cells and mouse models The findings show the potential of using NanoLiposomes for targeted gene editing in cancer The researchers conclude that this method could provide a new and effective way to treat prostate cancer particularly in patients requiring gentler treatment options This summary was initially drafted using artificial intelligence then revised and fact-checked by the author Indonesia — Nestled between a vast palm oil plantation and the Sumatran rainforest is Alue Buloh one must hike several kilometers down a mud and gravel road lined by land clearings and abandoned huts The residents of Alue Buloh seem to never run out of stories about their interactions and encounters with elephants “We can only accept our fate when the elephants ravage our crops,” said Junaidi a 42-year-old farmer who lives in Alue Buloh “The elephants will never forget their original route if we build a house or a farm within the elephants’ corridor Instead of killing or hurting the elephants for invading their farms the farmers provide them with space to feed and chase them away gently using loud noises — and in some cases Even as increasing habitat loss for the Sumatran elephant and poverty for the villagers complicate efforts to address the human-elephant conflicts local farmers are still exhaustively conducting voluntary patrols to protect the elephants and save their livelihoods These are the people living among the Sumatran elephants and working to conserve them along with the livelihoods of Cot Girek Junaidi heads the volunteer elephant patrol team in Cot Girek Protecting farmers’ livelihoods motivated him to join “Unlike government officials who receive a monthly salary we [farmers] can only rely on our crops for money,” he said He added he hopes for the government to intervene and help the farmers who have to deal with human-wildlife conflicts Junaidi said he’s encountered elephants from a very young age “Elephants are human too — they have strong feelings,” he said I remember seeing an elephant that fell into a canal and one of its mates immediately helped it Elephants typically roam the village at 3 in the morning “We use cannons and clanking noises to chase them away while saying ‘surut an activist from SAHARA (Yayasan Suara Hati Rakyat a local environmental NGO whose name translates into “the People’s Conscience Foundation”) is fighting the local culture of patriarchy through her activism in Cot Girek “Communities of women who live in secluded places that lack access [to education and information] such as Cot Girek are happy to discuss women’s rights,” she said The community is trained to grow a variety of crops such as rosella (Hibiscus sabdariffa) and improve their livelihoods and local economy with the use of non-timber forest products “The groups of women here are very focused on improving the local economy,” Husna said they lack acknowledgement when it comes to policymaking in the village.” Kelompok Meurah is led by Junaidi’s 61-year-old mother She said she’s tried many ways to repel the elephants from invading their houses and crops She recalled a time when Junaidi was young and a male elephant stood before him; she asked Junaidi to recite the adzan Maybe he would listen to us and love us.’ He recited it three times until the elephant moved back.” The role of women is crucial in protecting the village during elephant invasions While the men are away looking for and herding the elephants back into the jungle the women wait at home with torches and cannons I had to carry young Junaidi on my back while holding two torches in each hand to chase away the elephants,” Yasmin said we were all women protecting ourselves and herding the elephants back to the jungle.” a Kelompok Meurah member and the only female community member of the government’s Conservation Response Unit (CRU) said she and her team are always ready to deal with elephants entering the village They walk around the village toward the elephants to herd them back out to the jungle “We [the women] can’t just sit around in the house — we have to go out and help our friends herding the elephants,” Salamah said the team usually stands its ground to watch over the crops so the elephants can’t enter the farms They’re also ready to leave immediately to assist any family whose house is invaded by the elephants “The recent case in Alue Buloh was when someone’s house was completely destroyed by an elephant The elephant entered the house and rummaged in the fridge while the owners were away shopping for food in town,” she said a professional mahout from the Conservation Response Unit and was relocated to work with the CRU in Cot Girek since 2020 He said it’s their duty as mahouts to groom The government has formed 13 CRUs across Aceh province with trained elephants such as Marni to assist them in herding wild elephants back to the jungle Just like other captive elephants under the CRU’s care Marni knows whenever wild elephants are on their way to the village She raises her trunk to alert the residents of Cot Girek that the wild elephants are coming The mahouts and the volunteer elephant patrol team work together with the captive elephants to deal with the wild elephants and herd them back into the jungle “Elephants are intelligent; our elephants would follow us when we walk and come over when we call them,” Iwan said This story was produced with support from the Rainforest Journalism Fund in partnership with the Pulitzer Center This is the third installment in a three-part series about the Sumatran elephant. Read the first and second installments The “fortress conservation” model is under pressure in East Africa as protected areas become battlegrounds over history and global efforts to halt biodiversity loss Mongabay’s Special Issue goes beyond the region’s world-renowned safaris to examine how rural communities and governments are reckoning with conservation’s colonial origins and trying to forge a path forward […] but local people are rising to the challenge to preserve them Tashi Kunga stands before the Kag Choede monastery built into the Dhaulagiri mountain range on the Tibet-Nepal border The monk's carmine robes glint in the rain as he recounts the ancient legend of Guru Rinpoche's battle with a demon a demon wreaked havoc on a monastery in Tibet Guru Rinpoche chased it south to Upper Mustang in Nepal and defeated the demon following a ferocious battle burying the demon's remains across the mountain range The people of Mustang honoured the sacred grounds by building monasteries atop the demon's body parts This article in the Climate Guardians series was supported by funding from the European Journalism Centre through the Solutions Journalism Accelerator This fund is supported by the Bill & Melinda Gates Foundation "And right on the demon's heart, the capital of Lo Manthang [was built] in 1380," says Kunga, pointing towards the narrow alleys, ancient monasteries, and flat roofs adorned with prayer flags of one of the last medieval walled cities in the world have thrived in this remote region situated on top of the Tibetan Plateau One thing that has remained constant is the monasteries Increased rainfall saturates the rammed-earth buildings as moisture in the soil is drawn upward into the walls leading to issues such as leaking roofs and rising damp The emissions from travel it took to report this story were 70kg CO2. The digital emissions from this story are an estimated 1.2g to 3.6g CO2 per page view. Find out more about how we calculated this figure here. the paintings and the artifacts in the monasteries are embodiments of the gods themselves and we can't worship a half-damaged idol," says Kanga Buddhist monasteries have long been revered as the foundation of Tibetan culture serving as a vital hub for the creation and safeguarding of both tangible artifacts and profound intellectual traditions But as unprecedented weather patterns pose a threat to their cultural heritage local community members have stepped up to restore them from reinforcing walls to crafting metal statues and restoring paintings a team of local Lobas trained by Western art conservationists have replaced the old leaky roofs of the temples with round timbers sculpted pillars and the ceiling decorations giving these centuries-old monuments a new life Luigi Fieni, the lead art conservator at Lo Manthang Transforming a community of farmers into conservators has been challenging Most of the Lobas had never held a pen or a paint brush before and undertook extensive training before they began restoring the 15th Century paintings "But it all worked out," says Fieni "Tourists visiting Mustang were keenly interested in religion So we felt these sacred artifacts needed preservation not only for their historical significance but also for sustaining livelihoods here." although there was initial reluctance to accept any women in the group women are prohibited from touching sacred objects women did eventually take part in the Lo Manthang restoration project "It took years of discussion and negotiation with the local clergy and community but we succeeded in including local women in the wall-painting conservation team," says Fieni Married Tibetan women are typically responsible for cooking churning the butter and collecting yak dung for fuel When she received the opportunity to pursue new training and earn a daily wage in the restoration project "It enabled many of us [women] to break free from the limits of our homes and find new opportunities," she says confined to the corners with little purpose or prospects." wasn't there just to learn art or refine her painting skills She believed that by participating in the monastery's restoration she would earn spiritual merit and contribute to the promotion of Tibetan Buddhism through her work scraped-off outlines of deities and the vibrant expressions of revived figures you will recognise the incredible scale of everyone's efforts It has increased the cultural value of Lo Manthang," says Tsering we were able to play [a role] in it." leading to widespread flood damage in neighbouring villages The roads facilitating easy transport in the region have also contributed to rapid population decline with many Lobas moving to cities in search of economic opportunities Tashi Gurung stands out as a prime example of a farmer-turned-restorer who chose to remain in the region He wasn't exposed to Tibetan Buddhist art as a child but today he owns an art gallery in Lo Manthang His gallery showcases a remarkable array of thangkas and various deities and other spiritual beings such as the bodhisattvas His art sales not only sustain himself but also enable him to send his children to school Gurung acknowledges the profound impact of monasteries on his life "If it weren't for the monasteries I might have followed my friends who left Mustang for better opportunities abroad," he says The restoration efforts have sparked a cultural renaissance Numerous women and young people have received training in traditional craftsmanship long-forgotten festivals have been revived magnificent wall paintings restored for religious purposes and monasteries and nunneries are thriving once again "Om mani padme hum," the students at the Lo Monastic School chant the mantra that Tibetans use to invoke the bodhisattva of compassion the danger imperilling the walled city of Lo Manthang and its age-old monasteries doesn't stem from mythological demons but rather from the impacts of climate change we all need to join hands and save the monasteries because everything here spins around religion," says Tsering Seven images that transformed our world viewWatch how the maps and images of our planet from above have changed over the last two millennia. 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But that's – literally – only half of the story. all members of the Cumberland Valley Business Alliance recently had ribbon cuttings celebrating their openings In conjunction with the Cumberland Valley Business Alliance REVIVE MedSpa & Integrative Medicine celebrated their ribbon cutting at 2371 Buchanan Trail West on Tuesday who has been in practice for 17 years and in Franklin County since 2006 is a chiropractic physician focusing on specialized patient care Among their MedSpa services are hydrofacials and their Integrative Medicine services include physical medicine Class 4 laser treatments and electric stimulation “There is really nothing like REVIVE in this area Our services are unique to the area,” said Dr Learn more about REVIVE MedSpa & Integrative Medicine at ReviveMedSpa-Greencastle.com. They can be reached at office@revivemedspa-greencastle.com or by calling 717.263.9979.   REVIVE MedSpa & Integrative Medicine is a member of the Cumberland Valley Business Alliance which provides membership in the Greencastle-Antrim Chamber of Commerce and the Greater Chambersburg Chamber of Commerce In conjunction with the Greencastle-Antrim Chamber of Commerce celebrated their ribbon cutting and open house on Saturday Owners Jason and Karen Johns opened the shop in December of 2020 and sell locally grown beef and pork raised on their farm In addition to the deli area with fresh-made sandwiches the store offers a wide selection of fresh meats that are cut to client specifications.   Branded Meats & Deli. is open from 8:00 a.m. to 6:00 p.m.  Monday through Wednesday and  8:00 a.m. to 7:00 p.m. Thursday through Saturday.  More about Branded Meats & Deli, including daily specials, is available at www.facebook.com/brandedmeatsanddeli or by phone at (717) 593-9191 READ: Real estate firm opens Greencastle branch Family was the most important thing in his life and he watched college and professional sports of all kinds spending his free time hunting and fishing He loved animals and spending time with his grandchildren Dan was a lover of all things PSU and a staunch supporter of the legendary Joe Paterno Following a stint as a cryptanalyst with the NSA Olson took a job with IBM where he worked as a financial analyst for 26 years in a crash involving a lawn mower in Greene Township the public is invited to another ‘Paint Night With Patty’ in our downstairs area at Zion Reformed Church Penn State Extension Spring has sprung and while the change is welcome it is often when horticulturists see problems emerge that got The Franklin County Free Press is your local news website We provide you with the latest breaking news and videos straight from Franklin County PA The Franklin County Free Press is a Neil Publishing, LLC Any views or opinions expressed in this publication are of the individual or entity who submitted it and not the publication Our publication is open to the public to weigh in on matters of public interest AI technology assists in content creation on this site; verify independently for accuracy Call or Text: (717) 862-5656   Email: info@fcfreepress.com Please enable JS and disable any ad blocker Earth’s water may have been inherited from material similar to enstatite chondrite meteorites Tricycle is a nonprofit that depends on reader support A new photography book sheds light on conservation efforts in the hidden kingdom Carrying cables and Diva-Lites, landscape photographer Kenneth Parker takes another step up the mountain having previously braved the glaciers of Patagonia and the jungles of Laos and Myanmar but this 100-mile journey through the Himalayas takes the cake the air freezing and increasingly devoid of oxygen Parker is on his way to Lo Manthang, the fortress-like capital of Upper Mustang, to check out the work of conservator Luigi Fieni The so-called “Last Forbidden Kingdom,” today part of Nepal rose to prominence as a nexus for trans-Himalayan trade between Tibet This trade not only brought the kingdom earthly riches but also heavenly wisdom Inviting gurus from both sides of the mountain range Mustangi rulers turned their domain into a haven of Tibetan Buddhism They translated Sanskrit texts and built monasteries One of many projects taken on by Mustang’s last king—Jigme Dorje Palbar Bista who abdicated by order of the Nepalese government in 2008—was to work with the American Himalayan Foundation in an effort to restore many of the damaged temples which had fallen into disrepair over the last few centuries threatening the survival of the unique culture they represented Tibetan Buddhism is a Vajrayana lineage and includes four sub-schools–Nyingma These sects have a lot in common with other Buddhist traditions and the broader Buddhist landscape including the core beliefs of reincarnation the principle of nonviolence against all living beings the role of meditation as a means of reaching enlightenment as well as the commitment to the Buddha’s Four Noble Truths A key feature of Tibetan Buddhism—as well as other Mahayana Buddhist traditions—is reverence for bodhisattvas spiritual practitioners who are close to achieving enlightenment but delay its ultimate attainment to guide others along the path to Buddhahood having extinguished the flame of their being continue to be reborn for the benefit of all beings Instead of devoting themselves to a single Buddha Tibetan Buddhists worship a large pantheon of buddhas The faded wall paintings in Tubchen and Jampa depict this universe small details in the depictions of Buddhas and bodhisattvas—short necks and heart-shaped heads—betray influence from (and contact with) Nepal and other old Himalayan kingdoms like Guge Guru-devotion involves both your thoughts and actions The most important thing is to develop the total conviction that your Guru is a Buddha… If you doubt your Guru’s competence and ability to guide you your practices will be extremely unstable and you will be unable to make any concrete progress a mirror reflects both simply as they are—without judgment to otherwise distinguish between the two reds by attempting to hold the first The mirror stands imperturbable and immutable regardless of how we may otherwise perceive the circumstances as favorable or unfavorable.” the murals are not only historical artifacts but also objects of active worship—a duality that cannot and should not be ignored Fieni draws up a similarly passionate defense arguing that Western standards of conservation—derived from the study of “dead cultures” like ancient Rome and Egypt—cannot be applied to places like Upper Mustang where age-old ways of life have carried into the present I visited a small church with an old painting of St and there was room for local people to do what they wanted even if it took us a while to understand what that was I tried telling them what I had been told at school: that To leave the paintings incomplete is to make practice more difficult or nearly impossible which came to an end in 2019 when the Nepali government refused to renew his entry permit was an intellectual as well as spiritual experience he has since incorporated Buddhism into his worldview so that people can access it from different places,” he says it is not a matter of who is right and wrong but of accepting different choices because they relate to different cultures Since there can never be a globalization of conservation we must be open-minded and let other people proceed the way they feel is best for them.” with high altitude turning even the most mundane of tasks into demanding exercise “The scaffolding in the monasteries was three stories tall you had to go all the way up and down again I lost a lot of weight—10 or 20 kilos per summer.” But life was also simple are too often taken for granted: a warm bed the odd tourist sharing their prosciutto or Swiss cheese or not getting picked to clean the Porta-Potty at the end of the work the photographer quickly discovered a connection between his spirituality to his profession recounting the hardship of his Himalayan trek the better the picture.” Just as a seasoned Buddhist learns not to desire desirelessness so does Parker wait for photographs to come to him—sometimes for hours on end “There are no doctrines or words,” he says of Buddhism Tibetan Mustang: A Cultural Renaissance by Luigi Fieni and Kenneth Parker Images used with permission by Fieni and Parker Thank you for subscribing to Tricycle! As a nonprofit, to keep Buddhist teachings and practices widely available Δdocument.getElementById( "ak_js_1" ).setAttribute( "value" Already a subscriber? Log In Subscribe now to read this article and get immediate access to everything else Discover what over 30,000 subscribers have access to Full access to 30+ years of content with over 7,775 articles Continuous new writings from leading Buddhist teachers and New York Times bestselling authors Take an online Buddhism course at your own pace By Tony Koji Wallin-Sato By Mike Gillis Stephan Kunze in conversation with Jefre Cantu-Ledesma Please check your email to confirm your subscription Would you like to sign up for our other mailing lists By continuing, you agree to Tricycle’s Privacy Policy and Terms of Service Metrics details The mitochondrial calcium uniporter is a highly selective channel responsible for mitochondrial Ca2+ uptake The mitochondrial calcium uniporter shapes cytosolic Ca2+ signals Here using direct patch-clamp recording from the inner mitochondrial membrane we compare mitochondrial calcium uniporter activity in mouse heart heart mitochondria show a dramatically lower mitochondrial calcium uniporter current density than the other tissues studied mitochondrial calcium uniporter activity is barely detectable compared with that in other fly tissues As mitochondria occupy up to 40% of the cell volume in highly metabolically active heart and flight muscle low mitochondrial calcium uniporter activity is likely essential to avoid cytosolic Ca2+ sink due to excessive mitochondrial Ca2+ uptake low mitochondrial calcium uniporter activity may also prevent mitochondrial Ca2+ overload in such active tissues exposed to frequent cytosolic Ca2+ activity heart mitochondria manifested the lowest MCU activity The MCU current density in heart mitochondria was 30 times smaller than that in skeletal muscle The relevance of this evidence was further corroborated by our observation that mitochondria from the highly metabolically active Drosophila flight muscle manifests almost no MCU activity at variance with other populations of Drosophila mitochondria which indeed possess large current densities Our results are the first direct evidence that the amount of Ca2+ uptake mediated by the MCU varies between tissues The low mitochondrial Ca2+ conductance observed in tissues with a high mitochondrial volume density and that are continuously subjected to high cytosolic Ca2+ elevations such as mouse heart and Drosophila flight muscle is likely to be essential in having normal intracellular Ca2+ signaling as well as protecting these active tissues from toxic mitochondrial Ca2+ overload the success rate in formation of the ‘whole-mitoplast’ mode of the patch-clamp technique to record currents across the whole native IMM was low Here we patch-clamped mitoplasts derived from French-pressed mitochondria of mouse heart The French press protocol used to obtain vescicles of the IMM proved instrumental in increasing the yield of reliable and stable whole-mitoplast recordings from all tissues studied Whole-mitoplast current recorded before (black traces) and after (red traces) application of 100 μM Ca2+ to the bath solution Currents were elicited by a voltage-ramp protocol (shown above) from different mouse tissues (as indicated) Note that brown fat mitoplasts were isolated from mice deficient for uncoupling protein 1 (UCP1 Whole-mitoplast IMCU was normalized to the membrane capacitance (Cm) in all tissues examined representative transmitted differential interference contrast image of a mouse heart mitoplast obtained with French press Note the figure 8-shaped form of the mitoplast The lobe of the mitoplast containing only the IMM was less dense (white arrow) and clearly distinguishable from the lobe covered with the OMM (red arrow) Bottom right panel: Histogram showing average IMCU current densities in 100 μM Ca2+ in different tissues Current amplitudes were measured at 5 ms after stepping from 0 mV to −160 mV (see the voltage protocol) Note the low IMCU current density in heart compared with other tissues Statistical data are represented as mean±s.e.m (a) Left panel: representative whole-mitoplast current recorded from a mouse heart mitoplast before (black traces) and after (red traces) application of 150 mM Cl− to the bath solution Currents were elicited by a voltage-ramp protocol (shown above) The whole-mitoplast Cl− current was normalized to the membrane capacitance (Cm) but the traces represent an average of 30–50 original current traces to smooth out fluctuations of the outward Cl- current mediated by the large-conductance inner membrane anion channel (b) Left panel: representative whole-mitoplast current recorded from a mouse skeletal muscle mitoplast before (black traces) and after (red traces) application of 150 mM Cl- to the bath solution but the traces represent an average of 30–50 original current traces to smooth out fluctuations of the outward Cl- current mediated by the large-conductance anion channel (c) Histogram comparing average Cl− current densities in 150 mM Cl− in mouse heart (n=4) and skeletal muscle (n=3) Current amplitudes were measured at +80 mV (see the voltage protocol) and normalized to the Cm As skeletal muscle and heart are related tissues and showed the higest and lowest IMCU current densities we compared the biophysical properies of IMCU in these tissues to confirm that IMCU has the same properties in heart and skeletal muscle and is thus mediated by the same channel (a) IMCU elicited by a voltage-step protocol (as indicated on top ΔV=20 mV) in skeletal muscle (left panel) and heart (right panel) (b) IMCU recorded in the presence of 100 μM Ca2+ (blue trace) was completely inhibited by 50 nM RuR (red trace) both in skeletal muscle (left panel) and heart (right panel) The black traces indicate the baseline recorded in the absence of Ca2+ in the bath Currents were normalized to the membrane capacitance to compare IMCU in heart and skeletal muscle (a) Representative IMCU recorded in the presence of different concentrations of Ca2+ in the bath: nominal Ca2+-free (black) Representative IMCU recorded from a skeletal muscle mitoplast in the presence of 100 μM Ca2+ (red trace) Representative IMCU recorded from a heart mitoplast in the presence of 1 mM Ca2+ (red trace) (c) Histograms of the relative permeability of IMCU to Ca2+ Current amplitudes were measured at 5 ms after stepping from 0 to −160 mV (see the voltage protocol in b) This finding identifies the MCU as the Ca2+ channel with the highest known Ca2+ selectivity asked whether the MCU in mouse heart and skeletal muscle has the same high selectivity for Ca2+ over monovalent ions as the MCU in COS7 cells (a) Na+ current through the MCU under DVF conditions (black trace) and in the presence of either 11 nM (red trace) or 160 nM (blue trace) free Ca2+ in the bath solution Currents were normalized to membrane capacitance The voltage protocol is shown in bottom of the figure (b) The Na+ current through the MCU before (black trace) and after (red trace) the addition of 200 nM RuR to the bath solution (c) Monovalent current through the MCU in symmetrical 110 mM Na+ (black trace) and after replacement of bath Na+ with K+ (red trace) (d) Histogram showing average densities of Na+ current through the MCU in skeletal muscle (n=5) and heart (n=5) Current amplitudes were measured at 5 ms after stepping the membrane from 0 to −160 mV (see the voltage protocol) Thus IMCU in heart and skeletal muscle is likely mediated by the same molecule but heart mitochondria are endowed with dramatically lower MCU activity Although it has never been previously suggested one of the specific reasons for this may be a higher Ca2+ conductance of the IMM in neonatal cardiomyocytes as compared with adult cells Representative IMCU elicited in the presence of 100 μM Ca2+ (red traces) in a cardiac mitoplast isolated from a 2-day-old mouse The black trace indicates the baseline recorded in the absence of Ca2+ in the bath the same experiment performed with a cardiac mitoplast isolated from a 4-week-old (adult) mouse Currents were elicited by a voltage-ramp protocol shown above Whole-mitoplast IMCU was normalized to the Cm (b) Histogram comparing average IMCU current densities in 2-day-old (n=5) and adult heart (n=8) Current amplitudes were measured at 5 ms after stepping from 0 to −160 mV (see the voltage protocol) and normalized to the Cm we found a dramatic difference in the density of the mitochondrial Ca2+ current among Drosophila tissues (b) Left panel: Representative mitochondrial Ca2+ current recorded from a Drosophila flight muscle mitoplast in different concentrations of bath Ca2+: 5 mM (green) Right panel: Representative mitochondrial Ca2+ current recorded from a Drosophila not flight muscle mitoplast in the presence of different concentrations of Ca2+ in the bath: 1 mM (red) and 100 μM (blue) (c) Left panel: Histogram comparing average densities of Drosophila mitochondrial Ca2+ current at 100 μM in flight muscle (n=8) and not flight muscle tissues (n=3) Right panel: Histogram comparing average densities of Drosophila mitochondrial Ca2+ current at 1 mM in flight muscle (n=21) and not flight muscle tissues (n=6) In spite of the fact that the amplitude of the Ca2+ current was extremely low in flight muscle mitoplasts, these mitoplasts had very large Cl− current (Fig. 8) with the mean current density of 161±6 pA/pF at +80 mV, n=4. This is the first observation of the existence of a mitochondrial Cl− conductance in Drosophila. Left panel: representative whole-mitoplast current recorded from Drosophila flight muscle before (black traces) and after (red traces) application of 150 mM Cl− to the bath solution Right panel: Same experiment as in left panel but the traces represent an average of 10–20 original current traces to smooth out fluctuations of the outward Cl− current mediated by a large-conductance channel This uptake and the RuR-sensitive Ca2+ currents recorded from Drosophila mitoplasts are both likely to be mediated by a Drosophila homologue of the mammalian MCU the present consensus in the field is that all mammalian mitochondria are endowed with approximately the same MCU activity and the same high ability to accumulate Ca2+ this allowed direct studies of MCU activity under highly controlled conditions such as exact potential across the IMM and exact concentrations for all ions on both sides of the IMM the patch-clamp technique allows for the dissection of single mechanisms involved in mitochondrial Ca2+ transport and allows the characterization of MCU activity at the single organelle level limiting MCU activity in this tissue might not be necessary It must be noted that although singular heart mitochondria have a limited ability to uptake Ca2+ their abundance can result in combined Ca2+ accumulation comparable to other tissues it is not surprising that the MCU current density in Drosophila flight muscle is about four times lower than that in mouse heart some mitoplasts originating from Drosophila tissues other than flight muscle have very high MCU current density even exceeding the maximal MCU current density found in mice MCU activity varies dramatically between tissues Whether variations in MCU activity are due to tissue-specific regulation of the MCU (for example via auxiliary subunits) or to different tissue-specific expression of the MCU channel requires further investigation the tight control of MCU activity in highly active cells with intensive cytosolic Ca2+ signalling is likely crucial for the normal physiology of these cells and for the integrity of their mitochondria Misregulation of MCU activity in such tissues may lead to derangement of intracellular Ca2+ signalling and mitochondrial dysfunction followed by functional incompetence and degeneration and adult flies >5 days of age after eclosion were used Mouse or Drosophila mitoplasts (2–6 μm in size) were used for patch-clamp experiments and typically had membrane capacitances of 0.3–1.3 pF Currents were recorded using an Axopatch 200B amplifier (Molecular Devices) The voltage protocols and recording conditions are as indicated in the figures To minimize errors in the voltage clamp associated with the liquid junction potential a 3-M KCl-agar salt bridge was used as the bath reference electrode Gigaohm seals with mitoplasts were formed in the bath solution containing 150 mM KCl Capacitative transients were completely compensated right after the seal was formed Voltage steps of 350–500 mV from 15–50 ms were applied to rupture the IMM and obtain the whole-mitoplast configuration as monitored by the appearance of capacitance transients and an increase in baseline noise The access resistance and membrane capacitance of mitoplasts were determined using the Membrane Test tool of pClamp 10 (Molecular Devices) The averaged membrane capacitance of mitoplasts used in our experiments varied between tissues and was (0.65±0.04 pF The calculated voltage clamp errors associated with the access resistance never exceeded 10 mV Current amplitudes for histograms were measured within 5 ms after stepping the membrane from 0 to −160 mV Bath solutions containing 100 μM–1 mM Ba2+ or Mg2+ were obtained by the addition of a 1-M stock solution of the corresponding chloride salt into the HEPES-Tris solution without EGTA the following bath solution was employed: 110 mM K-gluconate tonicity adjusted to 300 mmol per kg with sucrose) All electrophysiological experiments were performed under continuous perfusion of the bath solution Data acquisition and analyses were performed using PClamp 10 (Molecular Devices) and Origin 7.5 (OriginLab) All electrophysiological data presented were acquired at 10 kHz and filtered at 1 kHz Statistical data are presented as the mean±s.e All general chemicals as were purchased from Sigma (USA) Activity of the Mitochondrial Calcium Uniporter Varies Greatly between Tissues Contributions of mitochondria to animal physiology: from homeostatic sensor to calcium signalling and cell death The interplay of mitochondria with calcium: An historical appraisal Role of calcium ions in regulation of mammalian intramitochondrial metabolism Decoding of cytosolic calcium oscillations in the mitochondria Beat-to-beat oscillations of mitochondrial [Ca2+] in cardiac cells The ups and downs of mitochondrial calcium signalling in the heart Mitochondrial calcium function and dysfunction in the central nervous system Mitochondria as all-round players of the calcium game A comparative study of the role of mitochondria and the sarcoplasmic reticulum in the uptake and release of Ca2+ by the rat diaphragm In vivo monitoring of Ca2+ uptake into mitochondria of mouse skeletal muscle during contraction Mitochondrial dynamics and their impact on T cell function mitochondria and reperfusion injury: a pore way to die Propagation of the apoptotic signal by mitochondrial waves Mechanisms by which mitochondria transport calcium The mitochondrial calcium uniporter is a highly selective ion channel A forty-kilodalton protein of the inner membrane is the mitochondrial calcium uniporter Integrative genomics identifies MCU as an essential component of the mitochondrial calcium uniporter MICU1 encodes a mitochondrial EF hand protein required for Ca2+ uptake MICU1 is an essential gatekeeper for MCU-mediated mitochondrial Ca(2+) uptake that regulates cell survival Mitochondrial transport of cations: channels Rapid changes of mitochondrial Ca2+ revealed by specifically targeted recombinant aequorin Measurements of mitochondrial calcium in vivo Calcium transport and porton electrochemical potential gradient in mitochondria from guinea-pig cerebral cortex and rat heart The relationship between free and total calcium concentrations in the matrix of liver and brain mitochondria Ultrastructural and biochemical studies of mitoplasts and outer membranes derived from French-pressed mitochondria Advances in mitochondrial subfractionation Patch-clamping of the inner mitochondrial membrane reveals a voltage-dependent ion channel 108-pS channel in brown fat mitochondria might Be identical to the inner membrane anion channel Evidence for the existence of an inner membrane anion channel in mitochondria A comparison of the effects of different divalent cations on a number of mitochondrial reactions linked to ion translocation Energy dependent bivalent cation translocation in rat liver mitochondria Ionic Channels Of Excitable Membranes 2nd edn Sinauer Associates (1992) Properties of Ca(2+) transport in mitochondria of Drosophila melanogaster Ca ion uptake by rat kidney mitochondria and its dependence on respiration and phosphorylation Elevated cytosolic Na+ decreases mitochondrial Ca2+ uptake during excitation-contraction coupling and impairs energetic adaptation in cardiac myocytes Ultrastructural quantitation of mitochondria and myofilaments in cardiac muscle from 10 different animal species including man Alterations of skeletal muscle in chronic heart failure Plasticity of skeletal muscle mitochondria: structure and function Morphometric study of early postnatal development of the thoracic aorta in the rat Cellular mechanisms of normal growth in the mammalian heart A quantitative and qualitative comparison between the right and left ventricular myocytes in the dog from birth to five months of age The effect of dietary restriction on mitochondrial protein density and flight muscle mitochondrial morphology in Drosophila Mitochondrial respiration in hummingbird flight muscles Mitochondrial dysfunction in Drosophila PINK1 mutants is complemented by parkin Synaptic mitochondria are critical for mobilization of reserve pool vesicles at Drosophila neuromuscular junctions A series of mutations in the D-MEF2 transcription factor reveal multiple functions in larval and adult myogenesis in Drosophila Download references This work was funded by the NIH Director’s New Innovator Award DP2OD004656-01 Present address: Present address: Department of Brain Science Daegu Gyeongbuk Institute of Science and Technology performed all electrophysiological experiments generated the Drosophila line expressing mitochondria GFP in the flight muscle All authors discussed the results and commented on the manuscript The authors declare no competing financial interests Download citation Sign up for the Nature Briefing newsletter — what matters in science where he dedicates a significant portion of his days examining schistosomiasis can carry a parasitic worm causing snail fever Schistosomiasis stems from a parasitic worm called Schistosoma japonicum The snails carrying these worms are found exclusively in 28 villages in Central Sulawesi specifically in Poso (Bada and Napu highlands) and Sigi districts (Lindu highland) Snail fever poses a persistent threat to the health of the 22 000 individuals residing in Poso and Sigi Districts Indonesia is the last country in the South-East Asia Region working towards eliminating schistosomiasis as a public health issue the 54-year-old Napu native with over three decades of experience in schistosomiasis reflects on the enduring impact of this parasitic disease on the local community as he pockets the snails and rides his trusty motorcycle back to the laboratory in Wuasa village The village's livelihood has been at risk for years prompting community-based measures such as reworking the land Cerni quickly geared up in his lab suit before placing the snail samples under the microscope sprays the host snail areas twice a year with niclosamide to eliminate snails The focus areas are surrounded by water seepage and long grass requiring thorough cleaning and trimming before spraying Cerni also works with the community to support screening and treatment \"There are instances where no one shows up when we go to collect stool samples Achieving the 80% target typically requires about a month in each village,\" he noted Cerni suspects that people's reluctance to be tested or undergo treatments is often influenced by weariness He shared the story of an individual who never received a positive schistosomiasis result in ten years and grew bored of the annual test find the treatments and test routines helpful and approach them with a positive attitude the mass drug administration (MDA) programme has consistently been received in a good light by the community – everyone Cerni Tolu analyzes human stool samples which are collected on a yearly basis. (WHO/Fieni Aprilia) Discovered under the microscope: the egg of Schistosoma japonicum in a stool sample Cerni packs away his microscopes and laboratory tools and performing lab tests with two other colleagues and some volunteers I can't say that I’m tired of these routines.\" Underscoring his dedication to eliminating schistosomiasis he emphasized the critical role of all members of the village to address this issue collectively WHO works with communities such as Cerni’s to move Indonesia closer to the goal of eliminating schistosomiasis once and for all.  The five-hectare land is one of the snail host focus areas several farmers have grouped together to transform the patch into productive ponds The combined efforts of the community bring about agricultural opportunities while protecting the village against snails "There are instances where no one shows up when we go to collect stool samples Achieving the 80% target typically requires about a month in each village," he noted I can't say that I’m tired of these routines." Underscoring his dedication to eliminating schistosomiasis a young farmer from Leubok Pusaka village in North Aceh district was on his way to his plantation when he spotted eight elephants on the riverbank One seemed to be staring at Saleh’s moving canoe Elephants!” Saleh and his friends shouted until all the animals were gone behind the trees a district near the northern tip of Indonesia’s Sumatra Island They couldn’t cross the river and they couldn’t return to the forest due to land-clearing activities in the opposite direction the herd finally managed to escape during a downpour the elephants found food in the villagers’ farms and destroyed four houses there’s been massive land clearing in North Aceh which lies along Sumatra’s eastern coast in the province of Aceh Despite the district’s enforcement of a moratorium on issuing new permits for corporate oil palm plantations conservationists report ongoing deforestation on the ground The North Aceh government has granted permissions for land clearing for smallholder oil palm farms some of which are said to be controlled by powerful people in the region has further fragmented the habitat of Sumatran elephants I believe the animals will soon go extinct,” Nurdin said doing their best to survive in the fast-changing environment “More than 85% of Sumatran elephants live outside conservation areas,” Azmi said there are four to five human-elephant conflicts reported every day conflict intensity escalated in North Aceh where much of the land has been cleared for oil palm a loud bang from a PVC air cannon woke Junaidi at 2 a.m The 41-year-old farmer heads the village’s elephant patrol team he knew it was a sign that wild elephants were moving in “Shoot the canon five times if you find wild elephants around your house” — that’s how the villagers have been told to communicate with others who might live kilometers away with poor cellular service but as patrol leader he had to get up and investigate despite the rain he walked some 10 kilometers (6 miles) along muddy roads around the village to check the situation Junaidi and other villagers in Cot Girek and Leubok Pusaka have been staying awake at night four wooden huts were reportedly destroyed by elephants a smallholder oil palm farmer who lives 3 km (nearly 2 mi) from Junaidi’s hut was shocked to see 400 oil palm shoots in his plantation chewed up by elephants who didn’t want her crops to meet the same fate an environmental activist from a local NGO called People’s Conscience said the increasing cases of human-elephant conflict are caused by habitat loss Cleared land can be seen from Junaidi’s hut Deforestation has eliminated the transition zone between the hills and the village “Elephants are coming from that hill,” Junaidi said pointing to a forested area over the horizon the geographical information system manager at Forest a conservation group focused on Sumatra’s Leuser Ecosystem North Aceh has long had one of the highest deforestation rates in Aceh province His analysis of satellite data generated by Planetscope which he called the most accurate satellite image provider shows the district lost 7,508 hectares (18,553 acres) of forest from 2017-2020 Satellite data generated from forest monitoring platform Nusantara Atlas show significant deforestation in Leubok Pusaka and Cot Girek said data he had collected from GPS collars tagged to elephants in North Aceh from 2016-19 showed that rainforest had been cleared within the elephants’ migration routes acknowledged land-clearing activities in Cot Girek “But these lands are classified as non-forest,” she said Lilis also said most of this clearing was done by local people rather than corporate actors the district has actively applied a freeze on new oil palm permits “We no longer give permit for companies to open up new oil palm plantations,” she said “Nor do we give oil palm seeds to smallholder farmers.” is the political extension of the movement It has always been poor villagers and elephants who suffer from conflict Junaidi’s hut sits between the forest and oil palm plantations owned by state-owned PT Perkebunan Nusantara I Junaidi said the company relies on the villagers’ patrol team to mitigate elephant conflicts “We are guarding their gate … all information about wild elephant movement comes from us,” he said (PTPN I did not respond to an interview request.) People like Junaidi and Saleh Kadri have to rely on their own resources to herd the elephants away from their village “We have reported about elephant conflicts in our village so many times but there has been no response from the government,” Saleh said We are tired of this … We hope the government can help farmers like us.” the elephants managed to leave the village They were last seen heading to Paya Bakung a subdistrict of North Aceh where a huge infrastructure project is being constructed To mitigate the annual flooding in Lhoksukon authorities are building the Kreung Keureto reservoir in Paya Bakung They will have to come back to … Cot Girek and finally Langkahan where they can’t cross the river and start their journey all over again,” Nurdin said “Poor elephants … they are chased from every side,” he added Banner image: A herd of elephants on the river bank in Cot Girek The principle guiding this roadmap is the understanding that the best available research evidence should be at the centre of the COVID-19 response and underscores the need to strengthen Indonesia’s health research system.  Rapid review dissemination from UGM CHPM-Cochrane on Incident Command System (under Pillar I: Country‑level coordination sourcing from national databases such as Garuda (Garba Rujukan Digital) and SINTA (Science and Technology Index) increase access to research articles and more effectively communicate findings to policymakers WHO and MoH along with other stakeholders joined together to cultivate an ecosystem that facilitates the utilization of evidence in the country One of the initiatives involved collaboration with experts from research institutions mandated to support COVID-19 response and preparedness at national and subnational level.   WHO Indonesia engaged over 20 experts from four institutions in capacity-building activities This initiative involved establishing a network of experts centred around synthesizing research evidence using rapid review methodologies and packaging the findings for effective use by policymakers.  Dr Bayu Dhite from UGM CHPM - Cochrane presenting rapid review findings on Incident Command System Aufia Espressivo from CISDI presented rapid review findings on community-based surveillance Each institution identified real-life public health and health system policy topics related to COVID-19 response and preparedness in terms of the structure organization and functions of the health system; actions and practices of stakeholders within the health system; or conditions and factors related to implementing certain changes Key insights from their research include:   Implementation of the Incident Command System (ICS): Highlighted the importance of ICS in managing health emergencies such as COVID-19 Strengthening non-communicable disease (NCD) health services: Strategic use of technology in NCD service delivery would enhance primary healthcare requiring increased investment in health information and digital skills.  Digital health surveillance: Identified the need for a technology innovation hub and matchmaking forum for developers and users.  Community-based surveillance (CBS): Leveraging Indonesia’s CBS system for tackling airborne diseases I Nyoman Sutarsa from School of Public Health Udayana presenting the effect of pandemic on NCD services through online platform The findings from the rapid reviews were deliberated during the 8th COVID-19 response intra action review (IAR) monitoring meeting held from 21-24 June 2023 in Jakarta These findings enriched and bolstered the discussions during the IAR meeting leveraging the lessons learned captured in the research evidence along the various COVID-19 response and preparedness pillars The network of experts will continue to provide support so that research evidence can be used more efficiently and effectively for enhanced evidence-based policymaking WHO will support Indonesia with development and establishment of a research council and accelerate the establishment of research databases to improve use of evidence These next steps will enhance Indonesia’s COVID-19 response and will lay the groundwork for a more resilient health system in Indonesia.  On a humid and rainy afternoon in Demak District Central Java health professional Dr Lisa Novipuspitasari looks out from her clinic and recalls the number of times that floods swallowed roads The coastal community she serves is familiar with the ferocity of water Heavy rains coupled with rising tides and coastal erosion make the area prone not just to flooding but the diseases it brings “Today I attended to another suspected leptospirosis patient in the emergency unit,” she shares with concern we escalated another case from observation in emergency care to inpatient treatment,” she adds Dr Lisa has been at the forefront of the fight against leptospirosis a zoonotic and climate-sensitive disease that can result in mild illness that may progress to a serious and sometimes fatal disease She is one of just a few leptospirosis specialists in Demak treating patients with symptoms ranging from flu-like illness to kidney failure and Severe Pulmonary Hemorrhagic Syndrome Dr Lisa standing in front of the RSUD Sultanfatah sign As one of just a few leptospirosis specialists she is leading efforts to prevent and control leptospirosis infections Leptospirosis barely registered as a concern even among health professionals,” she explains and with insights obtained from research by the National Research and Innovation Agency leptospirosis is misdiagnosed as dengue or typhoid leading to tragic outcomes when diagnosed and treated too late,” she says ruefully the Food and Agriculture Organization of the United Nations (FAO) USAID-IDDS and the Indonesian Ministry of Health and local government have joined forces to strengthen leptospirosis prevention and control in Demak which is expected to face rising sea levels and land subsidence of two to eight centimeters per year A leptospirosis patient undergoing treatment in Demak the Puskesmas is better prepared and capable of reducing deaths due to leptospirosis health and cross-sectoral partners such as the Coordinating Ministry on Human Development and Culture and the Ministry of Agriculture have conducted a joint risk assessment for leptospirosis developed a series of actionable recommendations to enhance preventive measures and also established a local zoonosis and emerging infectious disease coordination team in alignment with the “One Health” approach which recognizes the interconnectedness of animal these and other initiatives have helped strengthen leptospirosis surveillance and case detection enhance environmental management and increase rodent trapping knowledge and awareness-sharing among health workers and the public Amin supervises rodent trapping activities in the homes of leptospirosis survivors He has long recognized the correlation between leptospirosis cases and flood-prone areas where rodents that spread leptospirosis breed one of the rodent cadres in Bonang Village Amin says that knowledge obtained from Dr Lisa and other health workers at Bonang I Puskesmas has been crucial to strengthening preventive measures which are most effective when multiple activities are conducted concurrently with close communication between key actors and the public the One Health approach will continue to be central to WHO USAID-IDDS and Ministry of Health and local government efforts Dr Lisa reflects positively on the downward trend in leptospirosis cases in both incidence and severity – testament to the collective efforts of partners and the community to protect health and well-being from infectious diseases and the health impacts of climate change.   A type of meteorite called an enstatite chondrite has similar isotopic composition to terrestrial rocks and thus may be representative of the material that formed Earth. A new study published in the journal Science shows that these meteorites contain sufficient hydrogen to have delivered to Earth at least three times the mass of water in its oceans A piece of the enstatite chondrite meteorite Sahara 97096 Image credit: Christine Fieni / Laurette Piani / French National Museum of Natural History Enstatite chondrites are space rocks forged from the nebula that formed the Solar System They are rare, making up only about 2% of known meteorites in collections But their isotopic similarity to terrestrial rocks make them particularly compelling and the new study showed that their hydrogen and nitrogen isotopes are similar to Earth’s “Our discovery shows that the Earth’s building blocks might have significantly contributed to the Earth’s water,” said Dr. Laurette Piani a researcher at the Université de Lorraine “Hydrogen-bearing material was present in the inner Solar System at the time of the rocky planet formation even though the temperatures were too high for water to condense.” The building blocks of Earth are often presumed to be dry They come from inner zones of the Solar System where temperatures would have been too high for water to condense and come together with other solids during planet formation The meteorites provide a clue that water didn’t have to come from far away “The most interesting part of the discovery for me is that enstatite chondrites, which were believed to be almost dry, contain an unexpectedly high abundance of water,” said Dr. Lionel Vacher a postdoctoral researcher at Washington University in St “If enstatite chondrites were effectively the building blocks of our planet — as strongly suggested by their similar isotopic compositions — this result implies that these types of chondrites supplied enough water to Earth to explain the origin of Earth’s water The team also proposes that a large amount of the atmospheric nitrogen — the most abundant component of the Earth’s atmosphere — could have come from the enstatite chondrites “Only a few pristine enstatite chondrites exist: ones that were not altered on their asteroid nor on Earth,” Dr we have carefully selected the enstatite chondrite meteorites and applied a special analytical procedure to avoid being biased by the input of terrestrial water.” Coupling conventional mass spectrometry and secondary ion mass spectrometry allowed the scientists to precisely measure the content and composition of the small amounts of water in the meteorites it was commonly assumed that enstatite chondrites formed close to the Sun,” Dr “These chondrites were thus commonly considered dry and this frequently reasserted assumption has probably prevented any exhaustive analyses to be done for hydrogen.” Science 369 (6507): 1110-1113; doi: 10.1126/science.aba1948 Maryna Viazovska’s work on packing spheres and James Maynard’s solving of prime number conundrum honoured in Helsinki A Ukrainian mathematician who proved the best way to pack spheres in eight dimensions to take up the least space and an Oxford expert who has solved conundrums in the spacing of prime numbers considered the equivalent of a Nobel prize for mathematics presented at the International Mathematical Union awards ceremony in Helsinki have been announced as Prof James Maynard 35 of the École polytechnique fédérale de Lausanne of the University of Geneva and Institut des Hautes Études Scientifiques While the first Fields medal was awarded in 1936 since when it has been presented every four years to up to four mathematicians who are under 40 Viazovska, who was born and grew up in Kyiv, is only the second woman to receive the award, after the win by Iranian mathematician Maryam Mirzakhani who became a medalist in 2014 James Maynard: ‘Prime numbers are like the atoms for mathematicians.’ Photograph: Ryan CowanTalking to the Guardian from his hotel room in Helsinki Maynard – who is expecting the birth of his baby imminently – said he learned of his win while up a ladder tackling house renovations “I was grabbing my phone to use it as a torch to help to see if I had messed the painting up or not And I noticed that had I got a email then from the IMU president asking to have a zoom call,” he said Maynard’s citation points to his “spectacular contributions in analytic number theory” – among them his work on the distribution of prime numbers “Prime numbers are like the atoms for mathematicians,” said Maynard “In the same way that you can understand an awful lot about chemicals by knowing the atoms that make them up you can understand the huge amount about whole numbers and how they interact with multiplication – which turns out to be very important for things like cryptography – if you understand things about prime numbers.” Hugo Duminil-Copin Photograph: Matteo FieniA key step in trying to understand prime numbers is to look at the size of the gaps between them Maynard has made a number of breakthroughs including showing that sometimes prime numbers come unusually close together and sometimes unusually far apart said that when Maynard made an early pivotal discovery in how often pairs of prime numbers occur that are two steps apart – such as three and five – Graville told the young mathematician he must have made a mistake he’s not a one horse wonder … James has approached one [question] after another and just made massive headway.” Granville also praised the work of Viazovska who solved the problem of the densest way to pack spheres in eight dimensions and and relied on tens of thousands of lines of computer code Viazovska, said Granville, took the question even further, finding the solution in higher dimensions. “It turns out that in dimensions eight and 24, the solution is much easier than our common dimension, three,” Viazovska said in 2018 professor of mathematics at Princeton and also at the Institute for Advanced Study in Princeton “Viazovska invents fresh and unexpected tools that allow her to jump over natural barriers that have held us back for years,” he said involves the mathematical theory of phase transitions – for example when ice melts to liquid water – in statistical physics According Plus Magazine, Huh was more interested in poetry than maths at school, but became hooked on maths after attending lectures by Heisuke Hironaka Huh and colleague Petter Brändén found a connection between mathematical models for optimising situations involving variables that are related in a continuous way and those where the relationships are discrete “Finding this formal bridge was very satisfying.” Huh told the magazine “And what was even more pleasant for us is that once you have this bridge you can approach problems that were considered very technical and difficult in a very natural and easy way.” This article was amended on 6 July 2022 to amend a misspelling of Maryam Mirzakhani’s second name as “Mirzzakhani” Metrics details The newborn immune system is characterized by an impaired Th1-associated immune response Hepatitis B virus (HBV) transmitted from infected mothers to newborns is thought to exploit the newborns’ immune system immaturity by inducing a state of immune tolerance that facilitates HBV persistence we demonstrate here that HBV exposure in utero triggers a state of trained immunity characterized by innate immune cell maturation and Th1 development which in turn enhances the ability of cord blood immune cells to respond to bacterial infection in vitro These training effects are associated with an alteration of the cytokine environment characterized by low IL-10 and Our data uncover a potentially symbiotic relationship between HBV and its natural host and highlight the plasticity of the fetal immune system following viral exposure in utero To directly characterize the impact of HBV exposure on the newborn immune system we performed a comprehensive immunological analysis of the cord blood (CB) cells from neonates of HBV chronically infected mothers We report that HBV exposure in utero does not induce generic immunological defects but is associated with a mature immunological profile that enhances the ability of the neonatal immune cells to respond to unrelated pathogens in vitro (a) CB plasma cytokines were determined by multiplex assay in seven healthy controls and ten HBV+ mothers (b) Ratio of Th1/Th2 cytokine (IL-12p70/IL-10) in healthy and HBV-exposed CB P-values were calculated using Mann–Whitney test The frequency of total APCs (or HLA-DR+ cells) and of the various APC subsets was not affected by HBV exposure in utero (c) The mRNA expression (in Nanostring counts) of the chemokine CXCL13 in monocytes measured with Nanostring technology (d) Sorted CD14+ monocytes from healthy (n=4) and HBV-exposed (n=3) CB were incubated with 1 μg ml−1 ssRNA40 (TLR8 agonist) for 18 h and IL-12p40 in the supernatant measured using luminex (e) The median fluorescence intensity (MFI) expression of HLA-DR CD40 and CD80/CD86 on CD14+ monocytes from healthy (n=5) and HBV-exposed (n=10) CB (f) Sorted CD14+ monocytes from healthy (n=7) or HBV-exposed (n=8) CB were incubated with allogeneic CFSE-labelled CD3+T cells (E:T ratio 1) for 7 days and CFSE staining analysed using flow cytometry T-cell proliferation index was calculated using Flowjo Horizontal line in dot plots represents the median P-values in c–f were calculated using Mann–Whitney test Newborn T cells produce IL-8 but are defective in Th1 cytokine production11 As IL-12p40 has been shown to increase IFN-γ production in adult T cells we analysed the ability of CB T cells to produce Th1 and other important T-cell cytokines (that is (a) CB mononuclear cells were stimulated overnight with phorbol myristate acetate (PMA)/ionomycin and the cytokine production by CD3+T cells was measured using intracellular cytokine staining Dot plots show the percentages of cytokine-producing CD3+T cells from healthy (HC; n=6) or HBV-exposed (HBV; n=11) CB Cytokine production by CD3+T cells from the peripheral blood of pediatric and young adult patients with chronic HBV (HBV; n=10) and age-matched healthy controls (HC; n=33) were included for comparison (b) Representative FACS dot plots of Th1 cytokine (TNF-α IFN-γ and IL-2) production from healthy and HBV-exposed CB T cells after PMA/ionomycin stimulation double- and triple-producer Th1 cells and their respective percentages in mean±s.e.m (d) Percentage of T cells expressing the Th1 marker in CB of healthy (n=4) and HBV+ (n=4) mothers P-values were calculated using Mann–Whitney test.*P<0.05 We next analysed whether the enhanced immune maturation detected in HBV-exposed CB could result in a better ability of the neonatal immune cells to respond to unrelated pathogens CB mononuclear cells from healthy (n=3) or HBV+ (n=3) mothers were incubated with the bacteria P monocytogenes (multiplicity of infection (MOI) 1) for 18 h and the cytokine production in the supernatant was analysed using multiplex assay P-values were calculated using two-way analysis of variance and multiple comparisons were done using uncorrected Fisher’s least significant difference test (a) FISH analysis with X- and Y-chromosome-specific probes to detect maternal cells in male CB Graphs show the percentages of maternal cells (mean±s.e.m.) in CB unfractionated and fractionated cells (n=2 per group) Statistical significance between groups was calculated using Mann–Whitney test P-value <0.05 is considered statistically significant (b) Representative FISH images from each sorted cell population in healthy and HBV-exposed CB Nuclei were counterstained with DAPI (blue) Maternal cells (white arrow) contain green chromosome X signals and no Y signals (c) Quantitative PCR analysis using 48:48 Dynamic Arrays (Biomark System-Fluidigm) of CD14+ single cells derived from HBV-exposed CB of male neonate Two male (M)-specific genes (XKRY and TTY1) are expressed in CB cells and not in female (F) cells XIST gene is selectively expressed in female cells These results demonstrate that the immunological changes observed in HBV-exposed CB is unlikely to be due to an increase in maternal cell contamination but probably due to genuine maturation of the neonatal immune cells despite the latter being tested positive for HBV-DNA in the CB plasma (a) CD2− cells were enriched from CB using MACS beads and the presence of HBsAg was detected using immunofluorescent staining Detection of HBsAg in CB cells (white arrow) from healthy and HBV+ mothers (green–HBsAg; blue–DAPI) Images are representative of seven CB samples per group CB cells incubated with recombinant HBsAg was used as positive control (b) Percentage of HBsAg+ cells among DAPI+ cells in healthy and HBV-exposed CB (n=7 per group) P-value was calculated using Mann–Whitney test the presence of HBsAg+ cells in the CB of neonates of HBV+ mothers indicates that their immune system has been in contact with the virus or viral products before birth The low frequency of HBsAg+ immune cells detected in the CB of HBV-exposed neonates suggests that it is unlikely that HBV antigens can directly cause the maturation of monocytes in HBV-exposed CB We hypothesize that the altered cytokine environment detected in the CB plasma of HBV-exposed neonates could be responsible for the induction of monocyte/T-cell maturation CB mononuclear cells from healthy mothers (n=3) were incubated with rhIL-12p40 or rhIFN-α2 either alone or in different combinations of concentrations overnight and the activation phenotypes of T cells and monocytes were analysed by FACS Graphs (a,b) show the data from one representative sample Dotted lines indicate the basal level without any cytokine stimulation (a) The percentage of T-bet+ cells in CD4+ and CD8+ CB T cells (b) The median fluorescence intensity (MFI) of expression of HLA-DR (c) The activation phenotypes of T cells and monocytes in CB cells (n=2 denoted 1 and 2) stimulated with physiological concentrations of rhIL-12p40 (0.1 ng ml−1) and rhIFN-α2 (0.4 ng ml−1) no such evidence has been demonstrated so far in newborns during the course of a natural viral infection we demonstrated that HBV exposure in utero induces a state of trained immunity characterized by enhanced innate immune cell maturation and increased Th1 development this immune system maturation results in a better ability of the neonatal immune cells to respond to unrelated pathogen exposure the infant’s immune system does not only acquire a more pronounced Th1 T-cell profile but also decrease its ability to produce IL-10 and pro-inflammatory cytokines A further alteration induced by HBV exposure was the detection of higher levels of IFN-α2 that were only statistically significant in the CB plasma of Asian but not Caucasian HBV+ mothers Whether such differences could be explained by different HBV genotypes infecting the two cohorts (HBV genotypes B/C in Asian patients versus HBV genotype D in Caucasian patients) will require further analysis where virus-specific T cells can be detected in neonates A possible scenario is that HBV has evolved a special relationship with its human host: although the defective priming of HBV-specific T cells can predispose to HBV chronicity the induction of a trained immunity profile with a skewed Th1 response and suppression of pro-inflammatory events might have the advantage of decreasing mortality from exposure to unrelated pathogens but certainly a more precise evaluation of the immunological events that are occurring in the early phases of HBV infection is needed might actually be the source of IL-12p40 production in response to HBV Pregnancy is known to modulate the natural history of HBV infection but whether placental cells can actually play a direct role in the modulation of maternal or neonatal infections remains unknown All mothers in the HBV group were positive for HBsAg and negative for human immunodeficiency virus CB was collected from the umbilical vein using a direct dripping method into tubes containing heparin plasma was separated from whole blood and stored at −20 °C and CB mononuclear cells were isolated by density-gradient centrifugation on Ficoll-Hypaque The study in Singapore was approved by the Domain Specific Review Board at National University Hospital which was in accordance with the guidelines of the Singapore National Healthcare Group The study in Italy was approved by the Comitato Etico Azienda Ospedaliero Parma (Protocol 6274) and was in accordance with the guidelines of the Italian Minister of Health Blood samples from 10 pediatric and young adult CHB patients (12–30 years old) and 33 age-matched healthy controls used for the Th1 T-cell analysis were obtained from a viral hepatitis clinic at The Royal London Hospital Ethics approval was obtained from Barts and The London NHS Trust Ethics Review Board Monoclonal anibodies (mAbs) of anti-human-CD3-eFluor 605NC (clone OKT3 1:100) were obtained from eBioscience (San Diego 2.5:100) were obtained from Biolegend (San Diego anti-TNF-related apoptosis-inducing ligand-PE (RIK-2 6:100) were obtained from Becton Dickinson (BD 4:100) were obtained from R&D Systems (Minneapolis 6:100) was obtained from Molecular Probes (Carlsbad 2.5:100) was obtained from Beckman Coulter (Brea Live/Dead Fixable Dead Cell Stain Kits (yellow and aqua 1:1,000 in 1 × PBS) were obtained from Invitrogen 5 μM) were obtained from Invivogen (San Diego Phorbol myristate acetate (2 ng ml−1) and ionomycin (1 μg ml−1) were obtained from Sigma-Aldrich (Saint Louis HBV DNA was isolated from CB plasma or CB mononuclear cell lysates using the High Pure Viral Nucleic Acid Kit (Roche Applied Science) the cells were lysed in RLT buffer (Qiagen) and the lysate was passed through a blunt 20-gauge needle (0.9 mm diameter) fitted to an RNase-free syringe The lysate was centrifuged and the supernatant collected for HBV DNA extraction Five microlitres of internal control (HBV RG/TM IC) from the Artus HBV RG PCR kit (Qiagen) was added to the mixture of sample material and lysis buffer to control for the purification process HBV DNA was quantified using the Artus HBV RG PCR kit (Qiagen) on a Rotor-Gene Q platform The 95% HBV DNA detection limit of the assay was 20 IU ml−1 or 108 copies per ml Cells were cytospinned onto Superfrost Plus slides (Thermo Scientific) using CYTO-TEK Cytocentrifuge (Sakura Finetek) sealed with ProLong Gold Antifade Reagent with DAPI (4',6-diamidino-2-phenylindole; Invitrogen) and HBsAg staining was visualized using TissueFAXS system (TissueGnostics) The exposure time for fluorescein isothiocyanate (FITC) filter (for HBsAg staining) on the microscope was adjusted based on the negative controls (healthy CB cells) and positive controls (healthy CB cells incubated with recombinant HBsAg) to minimize autofluorescence/background staining without compromising signal The total number of HBsAg+ cells and the total number of DAPI-stained nuclei were manually counted in ten random high power fields ( × 20 magnification) CB mononuclear cells were washed in PBS and stained with Live/Dead Fixable Dead Cell Stain The cells were then washed in staining buffer (PBS Switzerland) and 0.1% sodium azide (Sigma-Aldrich)) stained for expressed cell surface molecules and analysed on a BD FACSAria or LSR II cytometer T-bet staining for Th1 cells was performed using Human FoxP3 Buffer Set (BD) Following overnight stimulation with phorbol myristate acetate (2 ng ml−1) and ionomycin (1 μg ml−1) in the presence of brefeldin A (2 μg ml−1) surface-stained cells were fixed and permeabilized (Cytofix/Cytoperm; BD) before being stained for produced cytokines Cells were then washed in staining buffer with 0.1% saponin (Sigma-Aldrich) before acquisition on LSR II cytometer Data were analysed using FACSDiva software (BD) APCs were sorted/gated based on lineage markers (CD3/CD7/CD56/CD19/CD20) and HLA-DR expression: CD14+ monocytes (lineage−HLA-DR+CD14+CD16low) and pDCs (lineage−HLA-DR+CD14−CD16−CD11c−CD123+) T cells were gated based on CD3+ expression a cutoff of two times the mean of the negative controls supplied in the kit was used to discriminate against nonspecific probe binding (noise) Samples were then normalized based on the geometric means of both the positive controls supplied in the kit and the panel of housekeeping genes of the normalized counts across all samples/mean normalized counts across all samples expressed as a percentage) of each gene was calculated and the mean coefficient of variation of the housekeeping genes was used as a cutoff to filter out genes that remain stable across all samples analysed Log2 normalized counts were used for clustering analysis Data were normalized (mean centering of genes) and hierarchical clustering of genes was generated using Cluster 3.0 (similarity metric: Euclidean distance clustering method: Average linkage) and visualized in TreeView Pan T cells (105 cells per well from a single healthy donor) were labelled with carboxyfluorescein succinimidyl ester (CFSE) and seeded in 96-well round-bottom plate with sorted CD14+ monocytes (ET=1) Pan T cells incubated with anti-CD3/CD28-coupled beads (Invitrogen 1:1 bead per cell ratio) were used as positive control cells were stained with anti-CD3-Horizon V450 (BD Biosciences) and acquired on a BD LSR-II flow cytometer T-cell proliferation was assessed by CFSE dilution Proliferation index was calculated using Flowjo software CB mononuclear cells were seeded in 96-well U-bottom plate at 105 cells per well in AIM-V media (Life Technologies) supplemented with 2% AB serum (Invitrogen) The number of monocytes was assumed at 10% of total cells baumanii were added to the culture at a multiplicity of infection 1 per monocyte and incubated overnight Cell supernatant was collected and analysed for cytokine production with the multiplex assay (Luminex) Cytokine concentrations in plasma samples and in supernatants of cultured cells were measured using MILLIPLEX MAP Human Cytokine/Chemokine Magnetic Bead Panel—Premixed 42 Plex (Millipore Analyte concentrations were determined by interpolation from a standard curve ELISA of IL-12 and IL-23 (R&D Systems) as well as of IFN-β and IFN-λ (PBL Assay Science) were performed according to the manufacturer’s instructions Healthy CB cells were seeded in 96-well U-bottom plate at 2 × 105 cells per well in AIM-V media (Life Technologies) supplemented with 2% AB serum (Invitrogen) NJ) were added either alone or in different combinations of concentrations for 24 h and the activation of T cells and monocytes analysed using FACS (BD) The concentrations of rhIL-12p40 tested were 0.1–1,000 ng ml−1 and 0.0004–4 ng ml−1 for rhIFN-α2 Nuclei were prepared for FISH analysis by resuspending the cells in 7 ml of 0.075 mol l−1 KCl and incubating them at 37 °C water bath for 15 min Two milliliters of 3:1 methanol:acetic acid was added to the cells centrifuged and the pellet was resuspended and washed twice with 7 ml of methanol:acetic acid solution Samples were stored at least overnight at −20 °C until slides were prepared Nuclei were dropped onto methanol-cleaned slides and air dried overnight on a 56 °C hot plate Slides pretreatment was performed in the following order: 1 × PBS at room temperature for 5 min 1% formaldehyde at room temperature for 10 min 1 × PBS at room temperature for 5 min and dehydrated in successive washes of 70% 80% and 100% ethanol at room temperature for 2 min each and allowed to air dry Poseidon Chromosome X and Y Satellite Enumeration Probes were obtained from Kreatech (the Netherlands) and used according to manufacturer’s protocol Nuclei were counterstained with DAPI solution Post-hybridization washes were performed as per manufacturer’s instructions Images were visualized and captured using the Isis Fluorescence Imaging System with the Nikon Eclipse 80i microscope The number of maternal cells were manually counted in 20 random high-power fields and expressed as a percentage of total nuclei All Raw Ct values were normalized to the assumed detection Ct level of 30 following recommendation from Fluidigm technical support heatmaps were produced using custom scripts and ggplot package in R The non-parametric Mann–Whitney U-test was used to determine the statistical significance of differences analysed the data and wrote the manuscript performed the experiments and analysed the data provided the samples and proofread the manuscript analysed the data and proofread the manuscript Accession codes: The Nanostring data reported in the paper have been deposited in the NCBI GEO database under accession number GSE65389 Trained immunity in newborn infants of HBV-infected mothers Immunosuppressive CD71 erythroid cells compromise neonatal host defence against infection Prevention of perinatally transmitted hepatitis B virus infections with hepatitis B immune globulin and hepatitis B vaccine Is a function of the secreted hepatitis B e antigen to induce immunologic tolerance in utero Age-dependent hepatic lymphoid organization directs successful immunity to hepatitis B Hepatitis B infection is associated with asymptomatic malaria in the Brazilian Amazon The transcription factor T-bet regulates parasitemia and promotes pathogenesis during Plasmodium berghei ANKA murine malaria Association of hepatitis B surface antigen carriage with severe malaria in Gambian children Preserved T-cell function in children and young adults with immune-tolerant chronic hepatitis B Interleukin-8 (CXCL8) production is a signatory T cell effector function of human newborn infants Innate immune function by Toll-like receptors: distinct responses in newborns and the elderly Unique efficacy of Toll-like receptor 8 agonists in activating human neonatal antigen-presenting cells A small jab—a big effect: nonspecific immunomodulation by vaccines Trained immunity: a memory for innate host defense Temporal analysis of early immune responses in patients with acute hepatitis B virus infection A longitudinal analysis of innate and adaptive immune profile during hepatic flares in chronic hepatitis B The RNA sensor RIG-I dually functions as an innate sensor and direct antiviral factor for hepatitis B virus IL-12p40: an inherently agonistic cytokine Profound lack of interleukin (IL)-12/IL-23p40 in neonates born early in gestation is associated with an increased risk of sepsis Direct multiplexed measurement of gene expression with color-coded probe pairs IL-21 is pivotal in determining age-dependent effectiveness of immune responses in a mouse model of human hepatitis B Pseudomonas aeruginosa infection in very low birth weight infants: a case-control study Maternal alloantigens promote the development of tolerogenic fetal regulatory T cells in utero A gene from the region of the human X inactivation centre is expressed exclusively from the inactive X chromosome Expression analysis of thirty one Y chromosome genes in human prostate cancer Detection of maternal cells in human umbilical cord blood using fluorescence in situ hybridization Hepatitis B virus translocates across a trophoblastic barrier The Developing Human: Clinically Oriented Embryology 7th edn W Persistence of infectious hepadnavirus in the offspring of woodchuck mothers recovered from viral hepatitis Mobilizing monocytes to cross-present circulating viral antigen in chronic infection Responsive-mode prophylaxis in the mealworm beetle A specific primed immune response in Drosophila is dependent on phagocytes Hemocyte differentiation mediates innate immune memory in Anopheles gambiae mosquitoes Evidence for macrophage-mediated protection against lethal Candida albicans infection Adaptive immune features of natural killer cells The role of BCG/PPD-activated macrophages in resistance against systemic candidiasis in mice Bacille Calmette-Guerin induces NOD2-dependent nonspecific protection from reinfection via epigenetic reprogramming of monocytes The immune tolerant phase of chronic HBV infection: new perspectives on an old concept Mature CD8(+) T lymphocyte response to viral infection during fetal life HIV-1-specific cytotoxic T lymphocyte responses in the first year of life Interferon-alpha as an immunotherapeutic protein Critical role for STAT4 activation by type 1 interferons in the interferon-gamma response to viral infection A unique mechanism for innate cytokine promotion of T cell responses to viral infections Correlation between vertical transmission of hepatitis B virus and the expression of HBsAg in ovarian follicles and placenta Herpesvirus latency confers symbiotic protection from bacterial infection Dating the origin and dispersal of hepatitis B virus infection in humans and primates IL-7 licenses activation of human liver intrasinusoidal mucosal-associated invariant T cells Download references We thank the subjects and their families for participating in this study Patrick Kennedy for providing the young adult samples Bindu Sukumaran for providing the bacteria strains John Conolly and Kaiting Ng for invaluable technical help with the Nanostring assay Komathi Paramasivam for assistance with the cell sorter and Veronica Khee for help with sample collection Emerging Infectious Diseases (EID) Program Azienda Ospedaliero-Universitaria di Parma UO Immunoematologia e Medicina Trasfusionale Supplementary Tables 1-2 and Supplementary Methods (PDF 13603 kb) Reprints and permissions Download citation Team Lead (NCD and Healthier Populations) WHO Indonesia presented fact sheets on the WHO-supported study on trans-fatty acids to Prof Vice Minister of Health of the Republic of Indonesia This moment marks the official launch of the results of the study The World Health Organization (WHO) has published the results from a ground-breaking study on the dietary sources of trans-fatty acids in Indonesia’s food supply This launch aims to support the issuance of government regulations to eliminate trans fats Trans fats or trans fatty acids are unsaturated fatty acids that come from natural or industrial sources Consumption of trans fats can significantly increase the risk of heart attacks and contributes to an estimated 500 000 deaths from coronary heart disease globally each year The baseline study involved laboratory testing of 130 products across four food categories: oils and fats packaged foods made with fats (such as biscuits cakes and bread) and prepared foods such as fried noodles WHO recommends trans fat levels in food of less than 2 grams per 100 grams of total fat almost 10% of the products surveyed or around 11 foods contained trans fat levels exceeding these recommendations High levels of trans fats are also found in popular and widely consumed snack products bakery products and street snacks such as martabak The highest concentration of trans fats is found in a mixture of margarine and butter which is 10 times higher than the WHO recommended limit it must be acknowledged that there is still a lack of data regarding trans fats in food The Ministry of Health really appreciates the efforts of WHO Indonesia to conduct a study of trans fat content in food,\" said the Vice Minister of Health of the Republic of Indonesia Dante Saksono Harbuwono at a high-level event in Jakarta The Vice Minister of Health Dante said that the most effective way to reduce trans fats in the food supply is through regulation WHO also encourages countries to adopt one of two best practice policies for eliminating trans fats The first step is to limit trans fat levels to 2% of the total fat content in all foods in 2018 WHO launched the global trans fat elimination initiative which calls on countries to eliminate trans fats globally by 2023 The REPLACE framework consists of six strategies: first reviewing trans fat dietary sources and policy landscape; second encouraging the replacement of trans fats with healthier fats and oils; third enact regulations to eliminate trans fats; fourth assessing and monitoring trans fat content in the food supply; fifth 53 WHO Member States have adopted best practice policies for eliminating trans fats including Denmark which became the first country to implement this policy since 2003 \"Denmark is the first country to ban trans fats in the food industry and this has been done since 2003 This can be done because before this ban their death rate from heart disease was very high,\" said the Vice Minister of Health Ten years after the regulation was implemented the death rate from heart and blood vessel disease fell by 20 percent The Vice Minister of Health Dante emphasized that the Government of Indonesia is also fully committed to implementing regulations prohibiting the use of trans fats in the food industry in Indonesia The Vice Minister of Health also assessed that limiting trans fats would reduce heart disease while saving Indonesia trillions of rupiah He hopes that the preparation of regulations involving multiple sectors can make Indonesia the next country to implement regulations according to good practices recommended by WHO “We will formulate these regulations in Indonesia people will be healthier so that the death rate due to heart and cardiovascular disease will decrease,\" said the Vice Minister of Health The Vice Minister of Health explained that the implementation of trans fat regulations would be accompanied by massive education especially in the informal sector such as small and medium traders 53 WHO Member States globally have adopted best-practice trans fat policies and WHO is working closely with the Government of Indonesia to ensure it becomes the next,” said WHO Representative to Indonesia “The release today of WHO’s baseline study marks an important step forward in improving the food environment for more than 275 million Indonesians enabling them to live longer and healthier lives,” he continued The Ministry of Health of Indonesia is dedicated to safeguarding the health and well-being of its citizens ensuring access to quality healthcare services Through collaborative efforts with stakeholders and communities nationwide the ministry strives to enhance healthcare infrastructure ultimately improving the overall health outcomes for all Indonesians Explore more about the Ministry of Health's initiatives and contributions at kemenkes.go.id or stay connected via their social channels Dedicated to the well-being of all people and guided by science the World Health Organization leads and champions global efforts to give everyone everywhere an equal chance at a safe and healthy life We are the UN agency for health that connects nations partners and people on the front lines in 150+ locations – leading the world’s response to health emergencies addressing the root causes of health issues and expanding access to medicines and health care keep the world safe and serve the vulnerable dr. Siti Nadia Tarmizi, M.EpidHead of Communication and Public Service Bureau, MoHkontak@kemkes.go.id+62 81281562620 Michael Vurens van EsCommunication Officer, WHOvurensm@who.int+62 81181101554 The Ministry of Health really appreciates the efforts of WHO Indonesia to conduct a study of trans fat content in food," said the Vice Minister of Health of the Republic of Indonesia "Denmark is the first country to ban trans fats in the food industry and this has been done since 2003 This can be done because before this ban their death rate from heart disease was very high," said the Vice Minister of Health people will be healthier so that the death rate due to heart and cardiovascular disease will decrease," said the Vice Minister of Health dr. Siti Nadia Tarmizi, M.EpidHead of Communication and Public Service Bureau, MoHkontak@kemkes.go.id+62 81281562620 Michael Vurens van EsCommunication Officer, WHOvurensm@who.int+62 81181101554 Fact sheet: Trans fatty acid in Indonesia study result (bilingual) Infographics: Trans fatty acid (in Indonesian) Step into the polio Supplementary Immunization Activity (SIA) in West Java where an inspiring collaboration between various organisations is taking place to protect children from polio and village officials join hands to ensure that every child in their communities has access to the novel oral poliomyelitis vaccine type 2 (nOPV2) they need conduct home visits to reach families who are unable to go to the integrated health posts (Posyandu) for polio vaccination they work tirelessly to ensure that every child is protected from poliovirus type 2 proving that the success of SIA is not just about the numbers but the commitment of the community Taken from 5-7 April 2023 in Purwakarta and Bandung this photo essay highlights the impact of cross-sectoral collaboration and the critical role of community involvement in achieving public health goals.  and the US CDC contributed to WHO Indonesia’s support for the polio outbreak response in West Java. Photos and words by Fieni Aprilia WHO Indonesia Digital Communication Officer  carefully transports a vial of nOPV2 in a carrier aboard his trusty motorcycle from Puskesmas Maniis to Sinargalih Village While many villages can be reached by vehicle Gugum and his colleagues often face rough terrain that requires them to travel on foot The single vial transported by Gugum will be used for 50 recipients protecting them from poliovirus type 2.  Maulana Yusuf (4) receives nOPV2 at the Sinargalih Village Hall in Sinargalih A total of 22 targeted recipientscame to the village hall that morning The vaccination session was followed by home visits to the nearby neighbourhood.  explain the importance of polio vaccination prior to administering the nOPV2 vaccine during a home visit in Sinargalih Village Uwais (9 months old) receives his nOPV2 vaccination while his mother Despite initial reluctance stemming from her husband's concerns Lita ultimately decided to vaccinate her children against polio thanks to the persuasive efforts of Minah and the health cadres of Sinargalih Village Uwais and Balqis received their first-ever vaccinations ensuring their protection against polio.  a team of midwives and health cadres from Sinargalih Village ventured out to visit a group of mothers and children who were spending time together these health workers made it their mission to ensure that every child in the community received their polio vaccination With their friendly and approachable demeanour the midwives and health cadres engaged with the parents and kids providing them with the information and resources they needed to stay healthy and protected Meet Noneng Jamilah (left) and Minah (right) two village midwives in West Java who are working tirelessly to ensure that every child in their community has access to life-saving vaccines who has been working at the Puskesmas since 2012 emphasises the importance of health workers checking the vaccination status of patients who visit for regular check-ups She administers necessary vaccines like the polio vaccine on the spot if needed Noneng also takes special care to ensure the comfort of children with disabilities during the vaccination process.  has been serving her community since 2006 and believes that working closely with health cadres and RT (Rukun Tetangga/Neighbourhood Associations) and conducting home visits are the most effective strategies to ensure that every child receives the vaccines they need She emphasises the importance of working collaboratively with village officials to ensure the smooth implementation of vaccination programmes Despite the challenges that arise from vaccine-hesitant parents who reject and intimidate them both Noneng and Minah remain committed to providing healthcare services to their community.  Both midwives also emphasise the importance of the Puskesmas’ monthly outreach programme to villages covering a wide range of topics from immunisation to stunting They mention the records kept in SIP books and how healthcare workers would check the records regularly before conducting home visits to children who may have missed their immunizations wears an END POLIO NOW pin on her backpack whilst assisting Minah (41) in their efforts to eradicate polio in Purwakarta.   Noneng and Minah worked together to administer vaccines to children during their door-to-door home visits.  Umar (4) and his mother Novi Handayani (27) show their polio vaccination card after they receive gifts from the Posyandu for receiving the vaccination in Posyandu Hapsari 2 "I came to the Posyandu because of my awareness of my child's development the Posyandu's monthly monitoring of my child's development is very good." said Novi Novi received information about polio vaccination directly from the Posyandu cadre Novi just learned about polio when giving Umar a vaccine at Posyandu Hapsari 2 - she admits that she did not seek information beforehand Posyandu is Novi's primary source of information for receiving information about health and child development.  The first Posyandu location at Bungursari Village The local government provides support in responding to the polio outbreak by conducting cross-sectoral socialisation in the district and cross-OPD (Organisasi Perangkat Daerah/Regional Government Organisation) to support Puskesmas in implementing the SUB PIN Polio programme Various corporate social responsibility (CSR) programmes also help with additional food and gifts for children who receive vaccines hospitals provide vaccine carriers to health centres that lack them.  Benjamin (2) receives nOPV2 polio vaccine as his mother stated that there were no challenges in providing the polio vaccine because of the high awareness level of the village community One way to help increase awareness is through polio socialisation.   and other health workers en-route to the next Posyandu post located inside a gated housing complex in Bungursari Village on 7 April 2023 Setting up two Posyandu spots was crucial to ensure that all children in the area receive their polio vaccination that day and maintain accessibility to everyone in the village.   administers the polio vaccine with precision and care Head of Disease Prevention and Control Division of Purwakarta Health Department ensuring a smooth and successful vaccination process for the community in Bungursari Village to ensure that the polio vaccination programme reaches all children the Purwakarta Health Department collects daily data from the Puskesmas and evaluates which areas have reached the target and which ones have not They then provide feedback to the health centres and conduct a sweeping activity if necessary "If there are children who have not been vaccinated due to illness we will wait until they recover before vaccinating them during the sweeping activity," Dr Matteo Fieni / International Mathematical Union is a Ukrainian mathematician who specializes in number theory She was awarded the 2022 Fields Medal in July for her work on finding the most efficient way to pack spheres of the same size in eight dimensions the Fields Medal is one of the highest honors in mathematics A professor at École Polytechnique Fédérale de Lausanne in Switzerland Viazovska received the medal and CAD15,000 (Canadian dollars) at a ceremony during the World Congress of the International Mathematical Union (IMU) She is the second woman to win the award in 86 years The only other female recipient was Maryam Mirzakhani (1977–2017) the same year as Brazilian mathematician Artur Ávila—the prize is awarded to up to four mathematicians under the age of 40 every four years the medal was awarded to 36-year-old Frenchman Hugo Duminil-Copin; 39-year-old South Korean–American June Huh; and 35-year-old Briton James Maynard © Revista Pesquisa FAPESP - All rights reserved offers some insights into his icon of bank I have lived in Lugano for the past forty years but the only public commission the city has ever entrusted to me was a bus stop,” Mario Botta humorously confesses This is despite the fact that this native of Switzerland is the first Swiss architect of international renown since Le Corbusier it’s no surprise that he describes Switzerland as both “a mother and a wicked stepmother.” He has built cathedrals yet his hometown has only ever commissioned him for a bus stop While Milanese writer Luca Doninelli called that very original creation—with lighting that changes according to the season—“the new cathedral of Lugano” counterbalancing in a way the architect’s bulkier private commissions in the Swiss canton of Ticino the bid he won in the early 1980s for the Banca del Gottardo in Lugano the architect would erect what he called a “mature structure,” his first major urban work located in Switzerland’s third largest financial center It has since become an international icon of bank architecture On the site of the bank had previously stood three turn-of-the-century villas and a parking garage all buildings familiar to Botta since his childhood tearing them down would not ultimately result in the destruction of his childhood memories “I didn’t want to do what most people were proposing We divided the site into four smaller buildings offset from each other by empty space to give it a dynamic look from the street a face that symbolizes the stratified memory of the city built up over time it’s rooted in the layering of history.” These words reflect the seminal influence of Louis Kahn one of Botta’s heroes and a former collaborator in Venice at the very beginning of Botta’s career The bank’s four facades are a take on the castle towers one finds in the nearby city of Como and the building’s Mediterranean grandeur earned it the nickname “Palazzo Botta.” Its precious architectural poetry derives from its two shades of natural stone a flourish that was anathema to bank architecture at the time with its rationally designed buildings and impersonal—empty—glass facades “The glass-and-metal banks of the ‘70s and ‘80s as today we only see them as buildings from the ‘70s without any identity of their own banks from the beginning of the century are archetypal Greek Doric temples and call to mind historical monuments.” Perhaps the most vividly iconic example of this style is to be found on Wall Street and in the Financial District of New York City Banking helped define Botta’s generation and it deserves a new interpretation: gone are the days of anonymous offices bustling with employees; rather banks should stand out boldly in the urban landscape Botta goes even further in his definition of the true identity of a bank: “We thought banks had to have sex appeal.” Though now owned by EFG International the bank is Switzerland’s fifth-largest private bank and the interior still exudes Botta’s great ambition The sacred aura of the place is undeniable which is no wonder since the architect has built iconic chapels through the Ticino region Yet he refrains from drawing a hasty parallel between banking and religion It’s a way to gild surfaces and truly generate space.” "We are the very first press outlet to be given permission to photograph it This is twenty-seven meters below lake level” works of art are not in the bank’s vaults but on its walls It is a kind of Guggenheim of the Swiss avant-garde The bank owns nearly one thousand of these marvels of Swiss nouvel art awaits us in the third basement of the building We are the very first press outlet to be given permission to photograph it “This is twenty-seven meters below lake level,” Martinoli whispers to us “The room is water safe to withstand up to two tons of pressure.” It’s one of the most beautiful vaults in the world; no vault of Swiss accounts before it has ever exuded such glamor Botta concludes the visit to his dulcet palace with: “It is a mysterious place that called for a kind of sacred feeling given its precious Deposits disappear and become mirrors in which we each find our own identity Get the latest news and updates straight to your inbox Paris Couture Fashion Week: A celebration of creativity Khaled Alrayes: Food critic turned entrepreneur Alice Temperley: A journey of passion and perseverance Kikuji Kawada's 'The Map / Visions of the Invisible' A partially collapsed cave containing the 12th to 14th century depictions of scenes from Buddha's life was unearthed last month by a team of Italian US and Nepalese conservators and archaeologists in Mustang a lost kingdom long forbidden to foreigners in the high Himalayas "Finding the cave was almost like a miracle," said Luigi Fieni a member of the team that used ice axes to cut its way into the inaccessible 3,400m-high cave in a region that for centuries was part of greater Tibet before being taken over by Nepal Foreigners were only permitted to enter Mustang in 1992 and Mr Fieni's team began work nine years ago restoring the spectacular wall paintings in a 15th century Tibetan monastery When they inquired about other art treasures in the region a villager remembered that as a boy he had seen a cave full of colourful paintings the Mustang cave paintings do not reveal a Tibetan but a strong Indian influence including the animals they depict - leopard the style evokes the fabulous cave paintings of Ajanta which predate the Mustang caves by several centuries." The location of the cave has been kept secret to deter art smugglers but the team call it "the snow leopard cave" as the animal's footprints were found inside "The cave paintings have been affected by wind and rain and really need restoration," Mr Fieni said and we're hoping now to raise funds for the project." The simultaneous discovery of ancient Tibetan manuscripts in nearby caves has led to speculation that the caves might have been a teaching retreat on the lines of the Buddhist university in Nalanda Mustang is of special significance to Buddhist experts because it is perhaps the only region where Tibetan culture and religion have survived over the centuries virtually untouched by time and modern Chinese colonisation They speak the Tibetan language; their origin is in the Tibetan culture," said Lama Guru Gyaltsen The opening up of the region has brought inevitable challenges to Mustang way of life Young men are leaving the tiny kingdom in search of work is certain to affect the tradition of rearing horses for transport and agriculture Building techniques using mud are likely to be abandoned in favour of modern construction methods.