Researchers have suspected for some time that the link between our gut and brain plays a role in the development of Parkinson's disease "Supplementation of riboflavin and/or biotin is likely to be beneficial in a subset of Parkinson's disease patients, in which gut dysbiosis plays pivotal roles," Nagoya University medical researcher Hiroshi Nishiwaki and colleagues wrote in their paper published in May 2024 The neurodegenerative disease impacts almost 10 million people globally, who at best can hope for therapies that slow and alleviate symptoms Symptoms typically begin with constipation and sleep problems up to 20 years before progressing into dementia and the debilitating loss of muscle control Previous research found people with Parkinson's disease also experience changes in their microbiome long before other signs appear Analyzing fecal samples from 94 patients with Parkinson's disease and 73 relatively healthy controls in Japan Nishiwaki and team compared their results with data from China While different groups of bacteria were involved in the different countries examined they all influenced pathways that synthesize B vitamins in the body The researchers found the changes in gut bacteria communities were associated with a decrease in riboflavin and biotin in people with Parkinson's disease Nishiwaki and colleagues then showed the lack of B vitamins was linked to a decrease in short-chain fatty acids (SCFAs) and polyamines: molecules that help create a healthy mucus layer in the intestines "Deficiencies in polyamines and SCFAs could lead to thinning of the intestinal mucus layer, increasing intestinal permeability, both of which have been observed in Parkinson's disease," Nishiwaki explains They suspect the weakened protective layer exposes the intestinal nervous system to more of the toxins we now encounter more regularly. These include cleaning chemicals, pesticides Such toxins lead to the overproduction of α-synuclein fibrils – molecules known to amass in dopamine-producing cells in the substantia nigra part of our brains eventually leading to the more debilitating motor and dementia symptoms of Parkinson's A 2003 study found high doses of riboflavin can assist in recovering some motor functions in patients who also eliminated red meat from their diets So it's possible that high doses of vitamin B may prevent some of the damage This all suggests ensuring patients have healthy gut microbiomes may also prove protective, as would reducing the toxic pollutants in our environment with such a complicated chain of events involved in Parkinson's disease not all patients likely experience the same causes so each individual would need to be assessed "We could perform gut microbiota analysis on patients or conduct fecal metabolite analysis," explains Nishiwak we could identify individuals with specific deficiencies and administer oral riboflavin and biotin supplements to those with decreased levels potentially creating an effective treatment." This research was published in npj Parkinson's Disease An earlier version of this article was published in June 2024 Metrics details We aimed to identify gut microbial features in Parkinson’s disease (PD) across countries by meta-analyzing our fecal shotgun sequencing dataset of 94 PD patients and 73 controls in Japan with five previously reported datasets from USA GC-MS and LC-MS/MS assays were established to quantify fecal short-chain fatty acids (SCFAs) and fecal polyamines α-Diversity was increased in PD across six datasets Taxonomic analysis showed that species Akkermansia muciniphila was increased in PD while species Roseburia intestinalis and Faecalibacterium prausnitzii were decreased in PD Pathway analysis showed that genes in the biosyntheses of riboflavin and biotin were markedly decreased in PD after adjusting for confounding factors Five out of six categories in carbohydrate-active enzymes (CAZymes) were decreased in PD Metabolomic analysis of our fecal samples revealed that fecal SCFAs and polyamines were significantly decreased in PD Genes in the riboflavin and biotin biosyntheses were positively correlated with the fecal concentrations of SCFAs and polyamines Bacteria that accounted for the decreased riboflavin biosynthesis in Japan and Germany were different from those in China1 different bacteria accounted for decreased biotin biosynthesis in the two country groups We postulate that decreased SCFAs and polyamines reduce the intestinal mucus layer which subsequently facilitates the formation of abnormal α-synuclein fibrils in the intestinal neural plexus in PD implying that ~50% of PD patients may follow this hypothesis Marked difference in gut microbiota across countries makes it difficult to identify causally associated intestinal bacteria and bacterial genes in PD patients We here performed shotgun-seq of fecal samples of 94 PD patients and 73 controls and meta-analyzed our dataset with five previously reported datasets to identify bacterial taxa and pathways that were specifically changed in PD across countries Decreased genes in fecal riboflavin and biotin biosyntheses were positively correlated with the decreased fecal SCFAs and polyamines different bacterial species were responsible for the decreased genes in fecal riboflavin and biotin biosyntheses in different countries a Left panel: PCoA plot of bacterial species in PD and controls in six countries with Bray–Curtis dissimilarity Circles and triangles indicate controls and PD Right panel: PCoA plot showing the center of gravity and the standard error of the overall compositions of bacterial species in six datasets in PD and controls with Bray–Curtis dissimilarity b Forest plot of α-diversities of bacterial species in six datasets FEM and REM represent fixed and random effect models The square sizes represent the number of samples Diamonds represent the mean and 95% confidence interval P-values of FEM and REM and I2 are indicated c The cumulative frequency plots of the relative abundance at the species level in six datasets The number of identified bacterial species in controls and PD in each dataset is indicated by a small circle at the end of the curve Orange and blue symbols represent increased and decreased CAZymes in PD Violin plots of (a) short-chain fatty acids and (d) polyamines in feces in controls and PD Concentrations are indicated by mg per g of freeze-dried fecal sample (mg/g FD) Median and interquartile range are indicated The sample sizes were 92 (PD) and 72 (controls) for acetate 91 (PD) and 70 (controls) for lactate and succinate and 86 (PD) and 66 (controls) for polyamines P-values were calculated by the Wilcoxon rank sum test Enzymes are indicated by EC numbers in squares Metabolites are indicated in green letters and open circles KEGG pathways are indicated by round squares Arrows indicate decreased or increased in PD Significant and non-significant changes are indicated by thick and thin arrows Forest plots of six datasets are indicated for the four significantly changed enzymes A right upper heatmap indicates the Spearman’s rank correlation coefficients between the four significantly changed enzymes and fecal SCFAs/polyamines a Heatmap showing the difference of the fractional CPMs between PD and controls of five most influential bacteria in each dataset b Forest plots of Faecalibacterium prausnitzii and Phocaeicola vulgatus in six datasets Supplementation of riboflavin to compensate for decreased riboflavin production by gut microbiota may be beneficial in PD patients concentrations of fecal polyamines may not be simply represented in plasma polyamines it is interesting to note that decreased spermine is a shared feature between plasma and feces in PD decreased riboflavin may lead to decreased polyamines through decreased vitamin B6 biotin may affect the SCFA and polyamine biosyntheses by serving as an essential cofactor biotin-dependent enzymes have not been extensively analyzed to date and we do not know which enzyme(s) become defective in the absence of biotin our analysis showed that the decreases of distinct sets of bacteria in PD in different countries led to a decrease of an identical pathway which poses another burden in the meta-analysis and the systematic review of gut microbiota at the genus and species levels these pathways were either up- or down-regulated without any statistical significance Solid arrows indicate the associations observed in the current study Broken arrows indicate speculations based on previous reports The sizes of the bacterial symbols represent the effects on biotin and riboflavin biosynthesis in which gut dysbiosis plays pivotal roles in PD We recruited 94 patients with idiopathic PD and their 73 healthy spouses who lived together from September 2015 to February 2018 Each PD patient was diagnosed according to the Movement Disorder Society’s (MDS) PD criteria We also excluded subjects who claimed to have taken antibiotics in the last four weeks The protocol was partially modified to use Lysing Matrix E Beads (MP Biomedicals USA) with FastPrep-24 5G (MP Biomedicals) for three cycles at 6.0 m/s for 60 s instead of vortex mixing Sequencing libraries were generated using NEBNext Ultra DNA Library Prep Kit for Illumina (NEB USA) according to manufacturer’s protocols and index codes were added to attribute sequencing fragments to each sample The library quality was assessed using TapeStation (Agilent Metagenomic sequencing was conducted using the HiSeq 2500 sequencing platform HiSeq Rapid SBS Kit v2 and HiSeq PE Rapid Cluster Kit v2 (Illumina USA) to obtain 3 Gbp per sample by 2 × 150 bp paired end-reads The total numbers of subjects including our own dataset were 813 PD patients and 558 controls The average read counts per sample ranged from about nine to sixty million reads The number of mapped reads per contig was normalized to CPM because a single EC number 2.1.1.43 constituted 25% or more of total CPM in 96 out of 156 samples (61.5%) the disease state (PD or control) was set to a covariate and the differences in bacterial abundances across datasets were set to batch effects PCoA was plotted with the function of vegdist in vegan (version 2.6-4) on R 4.2.3 using Bray–Curtis dissimilarity as a distance measurement and with the function of cmdscale in stats (version 3.6.2) on R 4.2.3 PERMANOVA was performed with the function of adonis2 in vegan (version 2.6-4) on R 4.2.3 a taxon or a CAZyme was considered to be significant when false-discovery rate (q-value) by Benjamini–Hochberg method for both FEM and REM was less than 0.05 that were significantly different between controls and PD in meta-analysis The difference between the USA and Japan was set to a random effect in the MaAsLin2 analysis The total number of samples analyzed with MaAsLin2 was 851 due to lack of the metadata P-values between controls and PD were normalized by the Benjamini–Hochberg method for each analysis of 73 EC numbers Statistical significance was set to q-value < 0.05 p-value of each pathway in each dataset was first calculated by GSEA An integrated p-value was then calculated for each pathway In the analysis of a KEGG pathway that was decreased in PD actual p-value was used when the pathway was decreased in PD in a dataset but p-value was set to 1.00 when the pathway was increased in PD in a dataset In the analysis of a KEGG pathway that was increased in PD a similar rule was applied in an opposite way A significance threshold of q-value by Benjamini–Hochberg method was set to 0.05 We previously established a GC-MS-based method to quantify fecal SCFA levels22 20 mg of FD feces was mixed with 1000 µL of 5 mmol/L NaOH and 300 µL of H2O The mixture was shaken vigorously with zirconia beads for 10 min and centrifuged at 13,200 × g for 20 min at 4°C The supernatant (333 µL) was mixed with 200 µL of H2O 50 µL hexanoic-6,6,6-d3 acid solution (internal standard) The resulting solution was then mixed for 1 min and centrifuged at 1900 × g for 5 min at room temperature Quantitative analysis was performed using an Agilent 7890A GC equipped with an Agilent 5975 inert mass spectrometer (Agilent Technologies) The precisions for all SCFA quantifications were less than 8.4% relative standard deviation (RSD) 20 mg of FD feces was mixed with 400 μL of H2O 10 μL of sulfosalicylic acid solution (1 mg/μL) and an internal standard solution (hexanoic-6,6,6-d3 acid) The mixture was shaken vigorously with zirconia beads for 10 min and centrifuged at 13,200 × g for 20 min at 4 °C The supernatant was then mixed with 20 μL of 6 mol/L HCl and 2 ml μL of diethyl ether and the upper layer was transferred into a new glass test tube The sample was dried by nitrogen gas at 40 °C The residue was dissolved in 300 μL diethyl ether After adding 20 μL N-tert-butyldimethylsilyl imidazole the derivatization reaction was performed at 60 °C for 30 min the solution was analyzed using GC-MS as described above The precisions for quantifications of lactate and succinate were less than 16% RSD and 2% RSD after derivatization using N-(9-fluorenylmeth-oxycarbonyloxy) succinimide which was composed of an Agilent 1200 infinity LC coupled with an Agilent 6430 Triple Quadrupole LC/MS System (Agilent Technologies) 1,6-diaminohexane was used as an internal standard The precisions for all polyamine quantifications were less than 4.9% RSD In order to examine the bacterial origins of nine significantly decreased EC numbers in “riboflavin metabolism” and “biotin metabolism” we calculated a fractional CPM contributed by each bacterium for each EC number the median of fractional CPMs in controls was subtracted from the median of fractional CPMs in PD to calculate ΔfCPM We assumed that a bacterium with the most negative ΔfCPM mostly accounted for the decreased EC number in a specific dataset the averaged ΔfCPMs were sorted in ascending order and the top five bacteria were selected Relative abundances of intestinal bacteria were analyzed by the Wilcoxon rank sum test for unmatched pairs with the mannwhitneyu functionality of scipy.stat on Python 3.11.2 the significance threshold for p-value after Bonferroni correction was set to less than 0.05 for both FEM and REM and the significance threshold of I2 was set to less than 40% the significance threshold for q-value by Benjamini–Hochberg method was set to less than 0.05 for both FEM and REM the significance threshold for q-values by Benjamini–Hochberg method was set to less than 0.05 Sequence data of our dataset are available at the DNA Data Bank of Japan (DDBJ) under the accession number of DRA016410 Gastric alpha-synuclein immunoreactive inclusions in Meissner’s and Auerbach’s plexuses in cases staged for Parkinson’s disease-related brain pathology Idiopathic Parkinson’s disease: possible routes by which vulnerable neuronal types may be subject to neuroinvasion by an unknown pathogen Vagotomy and subsequent risk of Parkinson’s disease Intestinal Dysbiosis and Lowered Serum Lipopolysaccharide-Binding Protein in Parkinson’s Disease Progression of Parkinson’s disease is associated with gut dysbiosis: Two-year follow-up study Short chain fatty acids and gut microbiota differ between patients with Parkinson’s disease and age-matched controls Meta-Analysis of Gut Dysbiosis in Parkinson’s Disease Short chain fatty acids-producing and mucin-degrading intestinal bacteria predict the progression of early Parkinson’s disease Gut microbiota are related to Parkinson’s disease and clinical phenotype Gut microbiota in Parkinson’s disease: Temporal stability and relations to disease progression Functional implications of microbial and viral gut metagenome changes in early stage L-DOPA-naive Parkinson’s disease patients Gut metagenomics-derived genes as potential biomarkers of Parkinson’s disease Cross-Sectional Study on the Gut Microbiome of Parkinson’s Disease Patients in Central China Association of Fecal and Plasma Levels of Short-Chain Fatty Acids With Gut Microbiota and Clinical Severity in Patients With Parkinson Disease Oral and gut dysbiosis leads to functional alterations in Parkinson’s disease Metagenomics of Parkinson’s disease implicates the gut microbiome in multiple disease mechanisms Integrated Multi-Cohort Analysis of the Parkinson’s Disease Gut Metagenome Metagenomics of the Gut Microbiome in Parkinson’s Disease: Prodromal Changes The gut microbiome of healthy Japanese and its microbial and functional uniqueness Multivariable association discovery in population-scale meta-omics studies Freeze-drying enables homogeneous and stable sample preparation for determination of fecal short-chain fatty acids Ueyama, J. et al. Effects of Pesticide Intake on Gut Microbiota and Metabolites in Healthy Adults. Int. J. Environ. Res. Public Health 20, https://doi.org/10.3390/ijerph20010213 (2022) The MetaCyc database of metabolic pathways and enzymes and the BioCyc collection of Pathway/Genome Databases Gut microbiome alpha-diversity is not a marker of Parkinson’s disease and multiple sclerosis A predictive index for health status using species-level gut microbiome profiling Riboflavin Has Neuroprotective Potential: Focus on Parkinson’s Disease and Migraine High doses of riboflavin and the elimination of dietary red meat promote the recovery of some motor functions in Parkinson’s disease patients Riboflavin Supplementation in Patients with Crohn’s Disease [the RISE-UP study] Regulation of immunological and inflammatory functions by biotin High doses of biotin in chronic progressive multiple sclerosis: a pilot study Targeting demyelination and virtual hypoxia with high-dose biotin as a treatment for progressive multiple sclerosis A metabolic profile of polyamines in parkinson disease: A promising biomarker Madeo, F., Eisenberg, T., Pietrocola, F. & Kroemer, G. Spermidine in health and disease. Science 359, https://doi.org/10.1126/science.aan2788 (2018) Gastrointestinal disorders in Parkinson’s disease and other Lewy body diseases Portincasa, P. et al. Gut Microbiota and Short Chain Fatty Acids: Implications in Glucose Homeostasis. Int. J. Mol. Sci. 23, https://doi.org/10.3390/ijms23031105 (2022) A New Mechanism of Biological Energy Coupling Soto-Martin, E. C. et al. Vitamin Biosynthesis by Human Gut Butyrate-Producing Bacteria and Cross-Feeding in Synthetic Microbial Communities. mBio 11, https://doi.org/10.1128/mBio.00886-20 (2020) Riboflavin Bioenriched Soymilk Alleviates Oxidative Stress Mediated Liver Injury and Gut Microbiota Modification in B(2) Depletion-Repletion Mice Riboflavin Supplementation Promotes Butyrate Production in the Absence of Gross Compositional Changes in the Gut Microbiota Two interconnected B vitamins: riboflavin and pyridoxine Riboflavin Is an Important Determinant of Vitamin B-6 Status in Healthy Adults A core gut microbiome in obese and lean twins The epidemiology of Parkinson’s disease: risk factors and prevention Symbiotic polyamine metabolism regulates epithelial proliferation and macrophage differentiation in the colon Increased intestinal permeability correlates with sigmoid mucosa alpha-synuclein staining and endotoxin exposure markers in early Parkinson’s disease Pesticides and Parkinson’s disease: Current and future perspective Microbial metabolite butyrate facilitates M2 macrophage polarization and function Macrophage Polarization: Different Gene Signatures in M1(LPS+) vs Population structure discovery in meta-analyzed microbial communities and inflammatory bowel disease using MMUPHin Extending and improving metagenomic taxonomic profiling with uncharacterized species using MetaPhlAn 4 Cochrane Handbook for Systematic Reviews of Interventions (John Wiley & Sons KEGG: kyoto encyclopedia of genes and genomes Urgent need for consistent standards in functional enrichment analysis Molecular hydrogen upregulates heat shock response and collagen biosynthesis and downregulates cell cycles: meta-analyses of gene expression profiles Rapid and Simultaneous Quantification of Polyamines in Human Plasma by LC–MS/MS After Pre-column Derivatization with N-(9-Fluorenylmethoxycarbonyloxy)succinimide Download references and Tomomi Yamada at the Nagoya University Graduate School of Medicine for their technical assistance We also acknowledge Division for Medical Research Engineering Nagoya University Graduate School of Medicine for their technical support on shotgun metagenomic analysis and Yusuke Okuno at the Nagoya City University Graduate School of Medicinal Sciences This study was supported by Grants-in-Aid from the Japan Society for the Promotion of Science (JP22K17343 to H.N. JP23H02794 to K.O.); the Ministry of Health Labour and Welfare of Japan (23FC1014 to K.O.); the Japan Agency for Medical Research and Development (JP22ek0109488 to K.O.) and the National Center of Neurology and Psychiatry (5-6 to K.O.) Center for Neurological Diseases and Cancer Nagoya University Graduate School of Medicine Department of Pathophysiological Laboratory Sciences Chubu University College of Life and Health Sciences HN performed bioinformatic analyses with the help of H.M All authors critically revised and approved the manuscript 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/s41531-024-00724-z Anyone you share the following link with will be able to read this content: a shareable link is not currently available for this article Sign up for the Nature Briefing newsletter — what matters in science The Industry's Leading Publication for Wineries and Growers It was April 23, 2011, a balmy and beautiful Petaluma spring day with temperatures in the mid-60’s, and Marcelo Aguero was competing in a tennis match at the Petaluma Valley Athletic Club, according to an account in the Petaluma Argus-Courier (https://www.petaluma360.com/article/news/grateful-cardiac-patient-thanks-for-saving-my-life/) As fate would have it Aguero collapsed in the middle of the match Aguero told the Petaluma Argus-Courier.   Aguero recovered and was out of the hospital by May 3 This week he was introduced as the new President and CEO of wine and spirits importer and supplier Kobrand Wine & Spirits Aguero’s first day on the job will be September 9 Aguero takes over for the retiring President and CEO Robert T who will be on the Management Committee of Kopf Family Enterprises the umbrella organization that includes Kobrand.  DeRoose and representatives from Jackson Family Wines and Kobrand for comment were unsuccessful.  a 52 year-old graduate of the University of Washington comes to Kobrand with over 30 years of experience in the wine industry including the last 20 years with Jackson Family Wines rising to Executive Vice President for Marketing and Direct to Consumer Sales before taking the top job at Kobrand He also held leadership positions as the Senior Vice President and General Manager at Santa Rosa the distributor owned by Jackson Family Wines Aguero was first hired by Jackson Family Wines in 2002 as its Northwest Regional Manager.  Aguero served as the Executive Sponsor for the Diversity and Inclusion program at Jackson Family Wines The Seattle native born to parents from Buenos Aires Argentina regularly speaks on issues of diversity including as recently as this year’s Unified Wine & Grape Symposium He becomes the first person of color to lead Kobrand in the company’s 80-year history.  It is with a figurative big heart that Aguero has pursued a career as a wine industry executive “The one thing that I appreciate about Marcelo is that he's a no-nonsense guy,” said Eric Mummert who worked with Aguero at Regal Wine Company from 2012-15 and recently left a position at Southern Glazer’s Wine & Spirits The worst bosses are the ones that don't have a sense of humor or who have a big ego that colors one's sense of purpose and how one deals with people.  He would be someone who I would work for again if he called me.”  Mummert expects Aguero will thrive in his new job immediately.  “Every good CEO should work in every different department to understand what is required to run a company and he's done all of that,” Mummer said “I can't imagine of a better candidate for the job He has the management skills.”  The career promotions and the latest job almost didn’t happen for the ambitious Aguero if the quick-thinking Giacomini hadn’t administered CPR immediately on the tennis court in 2011 Giacomini’s wife fetched a portable automated external defibrillator from the club Aguero’s doctor said that starting the defibrillator was key and preserved all his brain function is playing tennis again and thriving professionally.  DeRoose retires after 41 years with Kobrand including the last 15 as its President and CEO He came up through the Kobrand ranks and saw the company through international and domestic economic trauma.  “When we think of the cycles we have gone through other cycles that were tough in the industry that required flexibility and execution to get through those tough times Bob led Kobrand through those tough times,” said Dennis Cakebread co-owner and Chairman of the Board of the eponymous Cakebread Cellars What I would observe is that it is so unusual in the alcohol distribution businesses to have a relationship that long We've had a great relationship with Bob and Bob’s predecessors We go back to when he was the Estates Group manager and district manager I thought it was a good choice when they made him President he's continued to do the same thing and remained data focused.” Kopf with headquarters in New York City’s Empire State Building Kobrand was bequeath to his three daughters The company is now headquartered in Purchase less than three miles from the Connecticut border.  The Kobrand portfolio currently holds over 80 brands and producers of wine Producers include France’s Louis Jadot Italy’s Tenuta San Guido (producers of Sassicaia) Domaine Carneros and Mullan Road Cellars.  The Kobrand Portfolio is distributed by Republic National Distributing Company in 38 states.  “If you look at the portfolio and the brands they represent that speaks to a lot of things and it also speaks for their ability to work with so many different cultures,” Cakebread said “We are proud to be part of their portfolio.”  10 minutes by taxi from Nishiwakishi Station on the JR Kakogawa line AD Researchers have suspected for some time that the link between our gut and brain plays a role in the development of Parkinson's disease "Supplementation of riboflavin and/or biotin is likely to be beneficial in a subset of Parkinson's disease patients, in which gut dysbiosis plays pivotal roles," Nagoya University medical researcher Hiroshi Nishiwaki and colleagues write in their published paper While different groups of bacteria were involved in the different countries examined, they all influenced pathways that synthesize B vitamins in the body. The researchers found the changes in gut bacteria communities were associated with a decrease in riboflavin and biotin in people with Parkinson's disease BAUM made its debut as a global skin beauty brand in the prestige category It focuses on the “power of trees” that have been harmonizing with the changing environment since ancient times while promoting the initiatives to solve social issues and reduce environmental impact under the Shiseido’s philosophy “BEAUTY INNOVATIONS FOR A BETTER WORLD.” Three years after its conceptualization it was launched as a skin & mind care brand under the theme of “coexistence with trees.” What are the thoughts of the project team that endeavored to balance a sustainability mindset and the beauty business and then tackled the new unconventional challenge of using wood for cosmetics packaging to embody the theme We talked with three members: Ayami Nishiwaki Motoki Okoshi responsible for the outer packaging design at Global Innovation Center ―Could you talk about the background of launching BAUM As symbolized in the phrase “itadakimasu (thank you for the meal or offering),” maintaining a sense of awe and gratitude towards nature and living by efficiently circulating the limited resources in this island nation are deeply engrained elements of Japanese culture We started brand value development with that stance we ultimately decided to use the idea of ”coexistence with trees.” Trees are familiar materials for Japanese culture and symbolize the circulation of resources We think them the best motif to represent what a prestige brand should look like in the coming age ―I heard that your team focused on “a sense of familiarity upon use” as the consumers’ motive for selecting products while being aware of sustainability ―BAUM has been realized through solid building of the brand story that “consumers’ rich experience is organically connected with the context of being sustainable.” Am I right ―What kind of experience is the “skin & mind care” that BAUM touts ―Could you talk about the package design next How did you incorporate the themes like “sustainability” and “trees” ―You use upcycled wood from Karimoku Furniture for packaging ―There aren’t many cases of using wood for cosmetics packaging ―Had refill design been planned from the beginning as a means to create a sustainable brand ―What were the difficulties in producing actual packages How did you feel about the reactions of customers and society in general ―It was the passion of all the members that realized this unprecedented project through repeated trial and error This website is using a security service to protect itself from online attacks The action you just performed triggered the security solution There are several actions that could trigger this block including submitting a certain word or phrase You can email the site owner to let them know you were blocked Please include what you were doing when this page came up and the Cloudflare Ray ID found at the bottom of this page Sign up below to receive our newest workout routines With the right plan and the right discipline you can get seriously shredded in just 28 days "Big Bill" shares his wisdom to dominate one of the ultimate strength marks Follow these fit women we're crushing on for inspiration Those high-risk stunts apparently performed by Lucy Liu were really done by Michiko Nishiwaki She has been a stunt double for some big names in Hollywood as well as a fight choreographer After just missing being cast in Mortal Kombat Michiko decided to add stuntwoman to her resume She performed all the high-risk stunts in Kill Bill: Vol so in Japan being a woman in a muscle competition was positively a revolution Michiko Nishiwaki was able to find a gym and began training with weights in a quest to balance out her proportions and symmetry Michiko became the first female bodybuilding and powerlifting champion in Japan and held these titles for three consecutive years She opened three fitness clubs with her brother to capitalize on her competition success Being known as an expert in strength training she also did executive personal training and made connections with the elite of Japan Michiko forever changed the stereotype of the submissive and docile Japanese woman by becoming a strong role model for all Michiko’s acting career began in 1985 in Hong Kong Her first leading role was in the film My Lucky Stars Her performance caught the attention of Jackie Chan and Sammo Hung and soon she appeared with many of the best-known martial arts stars Michiko appeared in dozens of these genre films and then she had no problem learning to portray a martial arts expert on film which also included learning weapons training Michiko realized she could make use of her own gymnastic training like Jackie Chan whose background helped him develop a unique She eventually found herself in demand doing stunts as well as appearing as an actress and was quickly typecast playing villains in most of her Hong Kong films Michiko was brought to my attention by movie director Oliver Stone I scheduled a shoot with her in my studio for the Weider magazines Michiko is so popular around the world that I have seen my photos of her published all over the internet on sites devoted to martial arts She was not only a pioneer in the world of muscle & physique but an admirable role model in how she was able to capitalize on her athletic success and develop a career in movies Michiko Nishiwaki married and subsequently moved to Moorpark Having Michiko living close to Los Angeles gave me the chance to bring her to my studio to do photos for Muscle & Fitness I consider the images I created with her to be a significant addition to my photo archives and I’m happy to share them with magazine readers of the current era The big man will take to the stage at the 2025 Pittsburgh Pro The work behind the scenes is what separates champions from also-rans Here’s a IFBB pro's take on how to stimulate the medial deltoids Please view the main text area of the page by skipping the main menu. The page may not be displayed properly if the JavaScript is deactivated on your browser Japanese version Metrics details which functions as a t-SNARE component of the syntaxin18 complex is localized on the ER membrane and regulates retrograde transport from Golgi to the ER which generally releases pro-apoptotic proteins from Bcl2-mediated inhibition Previously we reported that retinal photoreceptors undergo BNip1-dependent apoptosis in zebrafish β-snap1 mutants we investigated physiological roles of BNip1-dependent photoreceptor apoptosis we examined the spatio-temporal profile of photoreceptor apoptosis in β-snap1 mutants and found that apoptosis occurs only during a small developmental window in which an apical photoreceptive membrane structure Transient expression of β-SNAP1 during this OS growing period prevents photoreceptor apoptosis in β-snap1 mutants enabling cone to survive until at least 21 dpf These observations suggest that BNip1-mediated apoptosis is linked to excessive activation of vesicular transport associated with rapid growth of the OS which inhibits protein transport to the OS rescued photoreceptor apoptosis in β-snap1 mutants which inhibits protein synthesis via the mTOR pathway also rescued photoreceptor apoptosis in β-snap1 mutants These data suggest that BNip1 performs risk assessment to detect excessive vesicular transport in photoreceptors These genes encode proteins with great functional diversity scaffolding of the apical photoreceptive membrane organelle known as the outer segment (OS) and transport of proteins to the OS through the connecting cilium it remains to be seen how dysfunction of these proteins triggers photoreceptor cell death After fusion of retrogradely transported vesicles on the ER membrane the syntaxin-18 cis-SNARE complex is generated This complex is normally disassembled by SNAP and NSF the syntaxin-18 cis-SNARE complex fails to be disassembled for lack of β-SNAP activity A possible mechanism underlying BNip1-dependent apoptosis is that BNip1 pro-apoptotic activity is suppressed by intramolecular binding of the N-terminal coiled-coil domain to the BH3 domain in the monomeric state; however when BNip1 forms the syntaxin-18 cis-SNARE complex BNip1 monitors reduction of β-SNAP activity and activates apoptosis when β-SNAP activity is decreased rescued photoreceptor apoptosis in coa mutants These data suggest that BNip1 performs risk assessment that detects excessive activation of vesicular transport in photoreceptors Bcl2-ER significantly rescues photoreceptor apoptosis in coa mutants EGFP-tagged Bcl2-ER was generated by replacement of the Bcl2 TM domain with a BNip1 TM domain (B) Retinas injected with mRNA encoding ER-mKO (magenta) and either mRNA encoding EGFP-tagged Bcl2 Fluorescent images shown in squares of upper panels and their green and magenta channels (middle panels) Bottom histograms indicate spatial profiles of EGFP and mKO signals along the line shown in the middle panels the spatial profile of EGFP-tagged Bcl2-ER signals correlates with that of ER-mKO signals whereas EGFP-tagged Bcl2 signals are not correlated (C) Retinas injected with mRNA encoding MT-mKO (magenta) and either mRNA encoding EGFP-tagged Bcl2 peaks of EGFP-tagged Bcl2-ER signals do not match those of MT-mKO signals whereas EGFP-tagged Bcl2 signals are broader than MT-mKO signals (D) Retinas of wild-type and coa mutant embryos combined with transgenic lines Tg[hs:mCherry-tagged Bcl2] or Tg[hs:mCherry-tagged Bcl2-ER] Tg+ and Tg− indicate transgenic and non-transgenic embryos Green color indicates zpr1 antibody signals INL and photoreceptor cell layer (PCL) are shown Right two columns indicate higher magnification of the PCL shown in the left two columns The interface between pigmented epithelium (PE) and the neural retina is shown as a white Arrowheads indicate rescued photoreceptors Scale: 50 μm (left two columns) and 10 μm (right two columns) (E) Percentage of zpr1-positive area relative to total retinal area increase the zpr1-positive fraction in coa mutants Bcl2-ER effectively rescues photoreceptor apoptosis in coa mutants Two-way ANOVA with the Tukey multiple comparison test apoptotic activity of BNip1 is activated on the ER membrane and overexpression of Bcl2 on the ER membrane effectively suppresses its apoptotic activity suggesting that Bcl2-ER is specifically located on the ER membrane These observations support the current model that lack of β-SNAP activity causes accumulation of the syntaxin-18 cis-SNARE complex which facilitates the interaction between the BNip1 BH3 domain and Bcl2 on the ER membrane in coa mutant photoreceptors Apoptosis occurs during OS growth stages from 60 to 96 hpf (A) Whole mount retinas of zebrafish transgenic embryos labelled with anti-red opsin antibody (magenta) Green and magenta colors indicate rod and cone photoreceptor OSs The OS appears at 60 hpf and progressively increases in size until 96 hpf (B) Whole mount retinas of zebrafish transgenic embryos labelled with zpr1 antibody (green) and TUNEL (magenta) Upper and lower panels indicate wild-type and the coa mutant genetic background a few TUNEL-positive cells were observed at each stage spread to the entire retina by 84 hpf and disappear by 96 hpf The lack of zpr1 and XP-GFP signals in coa mutants at 96 hpf suggest that almost all photoreceptors are eliminated by 96 hpf almost all photoreceptors undergo apoptosis from 60 to 96 hpf (C) Sectioned images of the photoreceptor cell layer labelled with anti-green opsin antibody (green) at 3 (D) Green cone OS size measured using section images shown in (C) The green cone OS increases linearly from 3 to 5 dpf One-way ANOVA with the Tukey multiple comparison test (E) Sectioned images of the photoreceptor cell layer labelled with fluorescent signals of Tg[XlaRho:XP-GFP] (green) at 3.5 The cone photoreceptor cell layer is counterstained with zpr1 antibody (blue) (F) Rod OS size measured using section at 3.5 The rod OS initially increases until 4.5 dpf is maintained as a plateau from 4.5 to 8.5 dpf and again starts to grow after 8.5 until 28 dpf One-way ANOVA with Dunnett’s multiple comparison test rod and cone apoptosis occur in coa mutants during their initial OS growth from 2 to 4 dpf β-SNAP1 activity after 48 hpf is required for photoreceptor survival at 84 hpf (A) Experimental design and results of heat-shock promoter-driven β-SNAP1 overexpression in coa mutants Zebrafish embryos were injected with a DNA construct that expresses mCherry-tagged β-SNAP1 under control of the heat-shock promoter Heat-shock treatment of injected embryos was performed at 36 and photoreceptor apoptosis was examined at 84 hpf Green or grey circles indicate rescue results for cone survival control embryos and coa mutant embryos injected with a DNA construct encoding hs:mCherry-β-SNAP1 at the one-cell stage and heat-shocked four times at 36/48/60/72 hpf Cone and rod survival was confirmed by labeling with zpr1 antibody (magenta) and fluorescent signals of Tg[XlaRho:XP-GFP] (green) Three or four heat-shock treatments effectively rescued photoreceptor degeneration in coa mutants (C) Percent zpr1-positive area relative to total retinal area Blue and red bars indicate wild-type and coa mutant embryos injected with DNA encoding hs:mCherry-β-SNAP1 Cone apoptosis was rescued by heat shock treatment at 36/48/60/72 hpf and 48/60/72 hpf which shows an equivalent rescue level to wild type (D) Percent XP-GFP-positive area relative to the total area of the photoreceptor cell layer Rod apoptosis was rescued by heat shock treatment at 36/48/60/72 hpf and 48/60/72 hpf Two-way ANOVA with Sidak’s multiple comparison test β-SNAP1 overexpression from 36 to 132 hpf is enough to maintain cones until 21 dpf but not rods in coa mutants (A) Experimental design of β-SNAP1 overexpression in coa mutants Heat treatments were applied at 12-h intervals from 36 to 132 hpf (red arrows) Green or grey circles indicate rescue results of cone survival and stage of analysis The blue arrow indicates the critical period for β-SNAP1-dependent cone survival (B) Retinas of coa; Tg[XlaRho:XP-GFP]; Tg[hs:mCherry-β-SNAP1] embryos or larvae that were heat-shocked from 36 to 132 hpf Wild-type with Tg[hs:mCherry-β-SNAP1] and coa mutant without Tg[hs:mCherry-β-SNAP1] are positive and negative controls Cone photoreceptors and rod OS were visualized by labeling with zpr1 antibody (green) and GFP signals from Tg[XlaRho:XP:GFP] (magenta) In coa mutants with Tg[hs:mCherry-β-SNAP1] photoreceptors were maintained until 21 dpf (C) Percentage of zpr1-positive area relative to total retinal area There was no significant difference in cone survival between wild-type embryos with/without Tg[hs:mCherry-β-SNAP1] and coa mutant embryos with Tg[hs:mCherry-β-SNAP1] at 6 the fraction of surviving cones in coa mutants with Tg[hs:mCherry-β-SNAP1] started to decrease although it was still significantly higher than that of coa mutants surviving cones markedly decreased in coa mutants with Tg[hs:mCherry-β-SNAP1] in a similar level to coa mutants (D) Higher magnification of photoreceptor cell layers indicated by squares of panel (B) The rod OS is increased in wild type during development the rod OS in coa mutants was similar to that of wild type at 6 dpf (E) Percentage of XP-GFP-positive area relative to total area of the photoreceptor cell layer the XP-GFP-positive fraction increases progressively during development in coa mutant embryos with Tg[hs:mCherry-β-SNAP1] the fraction is similar to that of wild type at 6 dpf significantly decreased after 10 dpf and became 0% at 28 dpf rod degeneration after 6 dpf is consistent with our model that BNip1-dependent photoreceptor apoptosis is linked to the OS growing period or β-SNAP2 is ectopically expressed in photoreceptors at later stages in coa mutants in compensation for loss of β-SNAP1 and prevents cone photoreceptor apoptosis cone photoreceptors survive after 6 dpf in the absence of SNAP activity in zebrafish these data suggest that BNip1-mediated photoreceptor apoptosis is specifically activated during the OS growth period Photoreceptor apoptosis and neuronal regeneration in coa mutants with overexpression of β-SNAP1 during the OS growth period (A) TUNEL of wild-type and coa mutant retinas with overexpression of β-SNAP1 during the initial OS growth period Yellow and red circles indicate TUNEL signals in photoreceptors of central retinas and retinal CMZ Higher magnification images of all TUNEL signals indicated by white squares are shown in the right side (B) The number of TUNEL-positive cells per 10,000 μm2 in photoreceptors and CMZs of 21-dpf wild-type siblings and coa mutants with overexpression of β-SNAP1 during the OS growth period TUNEL density is slightly increased in rescued coa mutant photoreceptors but does not differ significantly from that of wild-type siblings TUNEL density in the CMZ is significantly higher in coa mutants than in wild-type siblings (C) Anti-PCNA antibody labeling of wild-type and coa mutants with overexpression of β-SNAP1 during the initial OS growth period Arrowheads and arrows indicate rod progenitor cells and reprogrammed proliferative Müller cells the number of reprogrammed proliferative Müller cells was increased in the central retina In coa mutants with overexpression of β-SNAP1 during the initial OS growth period (bottom) the number of rod progenitor cells is markedly increased in the central retina; however reprogrammed proliferative Müller cells were only observed in the peripheral retina and not in the central retina (D) The number of PCNA-positive cells per section In coa mutants with overexpression of β-SNAP1 during the initial OS growth period numbers of proliferative Müller cells in both the CMZ (red bars) and the central retina (purple bars) were similar to that of wild type Rod progenitors in the CMZ were not significantly different from those of wild type (blue bars) rod progenitors in the central retina were twice as numerous as in wild type (green bars) Müller cells are not reprogrammed for neuronal regeneration but rod progenitors markedly increase in central retinas of coa mutants with overexpression of β-SNAP1 during the initial OS growth period but non-significant increase in apoptotic cell density in the surviving photoreceptor cell layer in coa mutants with overexpression of β-SNAP1 during the initial OS growth period at 21 dpf; however this subtle increase of apoptotic cell density may trigger proliferation of rod progenitor cells Blockade of intracellular vesicular transport rescues photoreceptor apoptosis in coa mutants (A) Retinas of 3.5 dpf wild-type and coa mutant embryos injected with standard MO Cone photoreceptors and rod OSs were visualized by labeling with zpr1 antibody (green) and fluorescent signals from Tg[XlaRho:XP-GFP] (magenta) In coa mutant retinas injected with either MO-ift88 or MO-kif3b cone photoreceptor degeneration was partially inhibited (B) Percentage of zpr1-positive area relative to total retinal area in wild-type and coa mutant embryos injected with standard MO and MO-ift88 MO-itf88 significantly rescued photoreceptor apoptosis in coa mutants (C) Percentage of zpr1-positive area relative to total retinal area in wild-type and coa mutant embryos injected with standard MO and MO-kif3b MO-kib3b significantly rescued photoreceptor apoptosis in coa mutants (D) Retinas of 3.5 dpf wild-type and coa mutant embryos treated with DMSO or rapamycin Lower panels indicate higher magnification of squares in upper panels The outline of the photoreceptor cell layer (white dotted lines) was visualized by labeling with zpr1 (green) and phalloidin (magenta) In coa mutant retinas treated with rapamycin the size of photoreceptor cell layer increased (E) Percentage of the photoreceptor cell layer relative to total retinal area in wild-type and coa mutant embryos treated with DMSO and rapamycin Rapamycin treatment significantly recovers the size of photoreceptor cell layer in coa mutants Since reduction of protein synthesis in the ER is likely to decrease vesicular transport to the OS these data suggest that decreased vesicular transport from the ER to the OS suppresses photoreceptor apoptosis in coa mutants These data support the possibility that excessive activation of vesicular transport activates BNip1 proapoptotic activity The alternative spliced form of xbp-1 and increased chop1 mRNA expression were observed only in tunicamycin-treated embryos (B) The ratio of the spliced form relative to the non-spliced form of xbp-1 mRNA We carried out three independent sets of PCR reactions for wild type The same set of PCR reactions are connected with the line The difference of coa mutant heads relative to wild-type heads and Tunicamycin-relative to DMSO-treated embryos in xbp1 spliced/non-spliced mRNA ratio were evaluated by Ratio paired t-test There was no significant difference between wild type and coa mutants (C) Ratio of chop1 mRNA relative to ef1α mRNA and Tunicamycin-relative to DMSO-treated embryos in chop1 mRNA expression were evaluated by Ratio paired t-test The role of BNip1-mediated apoptosis in photoreceptors (A) The current model for molecular mechanism of BNip1-dependent apoptosis BNip1 proapoptotic activity is inhibited by intra-moleculer interaction between N-terminal coiled-coil domain and BH3 domain BNip1 interacts with other components of the syntaxin-18 complex (Syntaxin18 Sec22b) through their SNARE domain and forms cis-SNARE complex in which BNip1 BH3 domain is activated to interact with anti-apoptotic Bcl2 leading to the activation of Bax-dependent apoptosis syntaxin-18 cis-SNARE complex is normally disassembled by SNAP and NSF syntaxin-18 cis-SNARE complex is abnormally accumulated on ER membrane which subsequently allows BNip1 to interact with Bcl2 (B) A model for physiological significance of BNip1-mediated apoptosis in photoreceptor differentiation and survival When intracellular vesicular transport is arrested subsequently triggering the ER stress response (Right panel) when intracellular vesicular transport is abnormally activated vesicular fusion events increase on target membranes along the secretory pathway Such excessive fusion events may trap SNAP molecules which reduces the relative contribution of SNAP to vesicular fusion on the ER membrane the syntaxin-18 cis-SNARE complex accumulates and the BNip1-dependent apoptotic pathway is activated (Left panel) Blockade of ciliary transport functions of Ift88 or Kib3b inhibits BNip1-dependent apoptosis BNip1 may cooperate with the ER stress response to maintain appropriate levels of vesicular transport in photoreceptors both cones and rods undergo apoptosis in β-snap1 mutants during their initial OS growth period from 2 to 4 dpf Although there is a second expansion phase of the rod OS it is likely that the initial expansion of the rod OS is enough to trigger rod apoptosis in β-snap1 mutants to determine the critical period of β-SNAP activity for photoreceptor survival we overexpressed β-SNAP1 during different time windows in β-snap1 mutants using the heat shock promoter Transient expression of β-SNAP1 from 36 to 132 hpf is enough to prevent cone and rod apoptosis in coa mutants at 6 dpf Since overexpressed β-SNAP1 protein can be maintained at least for 24 hpf after heat-shock treatment it is likely that cones are maintained in the absence of β-SNAP activity after 6 dpf rods failed to be maintained in the absence of β-SNAP after 6 dpf probably because rod OS genesis is still active from 8.5 to 28 dpf These data suggest that BNip1-dependent apoptosis is associated with OS growth of rod and cone photoreceptors All zebrafish experiments were performed in accordance with the Animal Care and Use Program of Okinawa Institute of Science and Technology Graduate School (OIST) which is based on the Guide for the Care and Use of Laboratory Animals by the National Research Council of the National Academies and has been accredited by the Association for Assessment and Accreditation of Laboratory Animal Care (AAALAC International) All experimental protocols were approved by the OIST Institutional Animal Care and Use Committee TUNEL was performed using an In Situ Cell Death Detection Kit (Roche) Rhodamine-conjugated phalloidin (Invitrogen R415) at 0.66 μM was applied to visualize actin filaments on cryosections labelled with zpr1 antibody Images were scanned using a confocal laser scanning microscope (Carl Zeiss BNip1 is localized in the ER membrane via its C-terminal TM domain2 we replaced the Bcl2 TM domain with a BNip1 TM domain EGFP or mCherry was fused to their N-termini To confirm that Bcl2-ER is localized in the ER we injected mRNA encoding EGFP-Bcl2-ER (200 ng/μL) with mRNA encoding monomeric Kusabira Orange (mKO)-tagged ER retention peptides (ER-mKO) (200 ng/μL) We also injected mRNA encoding EGFP-Bcl2-ER (200 ng/μL) with mRNA encoding mKO-tagged mitochondrial-located peptides (MT-mKO) (400 ng/μL) We also generated DNA constructs expressing mCherry-Bcl2 and mCherry-Bcl2-ER under control of the heat shock promoter We established zebrafish transgenic lines Tg[hs:mCherry-tagged Bcl2] and Tg[hs:mCherry-tagged Bcl2-ER] Morpholino was dissolved in water at 250 μM for MO-ift88 and injected into wild-type embryos at the one-cell stage In accordance with a previous report36 wild-type and coa mutant embryos were treated with rapamycin (LC laboratories Photoreceptor survival was evaluated with rhodamine-conjugated phalloidin labeling the photoreceptor cell layer was outlined and its size was determined Means and standard deviations were calculated Statistical analysis was done using two-way ANOVA with the Tukey multiple comparison test Total RNA was prepared from 2.5-dpf wild-type and coa mutant embryos Complementary DNA strands were synthesized from total RNA using 3′RACE System for Rapid Amplification of cDNA Ends (Invitrogen Cat#18373-027) and used as a template for semi-quantitative PCR and ef1α cDNAs were amplified with TaKaRa Ex Taq (TaKaRa Biochemicals) ER stress-induced wild-type samples were obtained by treatment with Tunicamycin (Nacalai code 35638-74 Amounts were quantified using ImageJ (NIH) Statistical analysis employed two-tailed Ratio paired t-tests Molecular genetics and emerging therapies for retinitis pigmentosa: Basic research and clinical perspectives The BH3-only SNARE BNip1 mediates photoreceptor apoptosis in response to vesicular fusion defects a family of NSF attachment proteins involved in intracellular membrane fusion in animals and yeast SNAP family of NSF attachment proteins includes a brain-specific isoform A fusion protein required for vesicle-mediated transport in both mammalian cells and yeast The mechanisms of vesicle budding and fusion Adenovirus E1B 19 kDa and Bcl-2 proteins interact with a common set of cellular proteins Involvement of BNIP1 in apoptosis and endoplasmic reticulum membrane fusion Functional identification of the apoptosis effector BH3 domain in cellular protein BNIP1 The unfolded protein response and cell fate control Intraflagellar transport genes are essential for differentiation and survival of vertebrate sensory neurons Kinesin-2 family in vertebrate ciliogenesis Mutation of DNA primase causes extensive apoptosis of retinal neurons through the activation of DNA damage checkpoint and tumor suppressor p53 The role of subunit assembly in peripherin-2 targeting to rod photoreceptor disk membranes and retinitis pigmentosa The 1D4 antibody labels outer segments of long double cone but not rod photoreceptors in zebrafish Early onset of phenotype and cell patterning in the embryonic zebrafish retina The development of photoreceptors in the zebrafish Cloning and characterization of six zebrafish photoreceptor opsin cDNAs and immunolocalization of their corresponding proteins Targeted effects of retinoic acid signaling upon photoreceptor development in zebrafish Retinoic acid alters photoreceptor development in vivo Late-stage neuronal progenitors in the retina are radial Muller glia that function as retinal stem cells Stem cells in the teleost retina: Persistent neurogenesis and injury-induced regeneration Genetic evidence for selective transport of opsin and arrestin by kinesin-II in mammalian photoreceptors is essential for vertebrate photoreceptor assembly and maintenance A transgenic zebrafish model for monitoring xbp1 splicing and endoplasmic reticulum stress in vivo Transgenic zebrafish model to study translational control mediated by upstream open reading frame of human chop gene Comparison of topographical patterns of ganglion and photoreceptor cell differentiation in the retina of the zebrafish How variable clones build an invariant retina Cone photoreceptor types in zebrafish are generated by symmetric terminal divisions of dedicated precursors The Zebrafish Book: A Guide for the Laboratory Use of Zebrafish (Brachydanio rerio) TNFalpha induces muller glia to transition from non-proliferative gliosis to a regenerative response in mutant zebrafish presenting chronic photoreceptor degeneration maintenance of forebrain compartments and patterning of retinal neurites Functionally conserved effects of rapamycin exposure on zebrafish Download references Mamoru Fujiwara supported genotyping and histology experiments This work was supported by a grant from OIST to I.M Okinawa Institute of Science and Technology Graduate University Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations Download citation DOI: https://doi.org/10.1038/s41598-020-74360-x Sorry, a shareable link is not currently available for this article. 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Volume 8 - 2021 | https://doi.org/10.3389/fmed.2021.812370 The mean percentage change in newly started clinical trials for diseases other than COVID-19 between each month in 2019 and the corresponding month in 2020 was −7.5% with the maximum of −57.3% observed between April 2019 and April 2020 the mean percentage change of reported results for each month in 2019 and 2020 was −5.1% with the maximum of −27.4% observed in July 2020 The activity of clinical trials was decreased as the number of COVID-19 patients was increased and a statistically negative correlation was observed between the prevalence of COVID-19 and the percentage decrease in the number of clinical trials stared or reported results decreases were observed in the latter half of 2020 compared with the same period during the previous year A considerable decline in non-COVID-19 activity for all indicators regarding clinical developments was suggested during the first wave of the COVID-19 pandemic It is important to recognize the situation and continue to make efforts to conduct clinical trials for both COVID-19 and no-COVID-19 for new medical developments in the future Crucial clinical trials are being conducted internationally which can lead to new drug applications in each participating country or region medical development for non-COVID-19 is also important this study mainly focused on clinical trials for non-COVID-19 The number of clinical trials for diseases other than COVID-19 was calculated by subtracting the number of clinical trials for COVID-19 (entering “COVID-19 OR SARS-CoV-2” in the “Condition or disease” field) from the number of all clinical trials Monthly data regarding reported results were extracted by specifying the date using “Results First Posted” in the advanced search studies labeled with the status “Completed” were identified as completed clinical trials with results The total number of COVID-19 cases was obtained from the website “Worldometer Coronavirus” (9) The total number of COVID-19 cases at the end of the previous month was used to examine the impact on clinical research activities during that month extracted from the cumulative graph of total cases by country (A) Number of newly started clinical trials for diseases other than COVID-19; (B) Percentage change of newly started clinical trials for diseases other than COVID-19 Relationship between total cases of COVID-19 and percentage change of the number of newly started clinical trials for diseases other than COVID-19 in regions with more than 30 studies The size of the bubbles reflected the number of studies The number of clinical trials for diseases other than COVID-19 that were started during the fourth quarter of 2020 showed signs of recovery but this was due to the increase in the number started in the United States the number of clinical trials started for diseases other than COVID-19 was still less than that in during the same month of the previous year in regions other than the United States the total number of clinical trials (COVID-19 and non-COVID-19) recovered rapidly and has exceeded the number in the same month of the previous year since June 2020 (A) Number of clinical trials for diseases other than COVID-19 with results; (B) Percentage change of clinical trials for diseases other than COVID-19 with results Relationship between total cases of COVID-19 and percentage change of the number of clinical trials with results in regions with more than 30 studies A similar tendency was observed in the number of completed clinical trials with results in 2020. The mean number of completed clinical trials with results for diseases other than COVID-19 in 2020 was 373 (SD, 38). The mean percentage change for each month between 2019 and 2020 was −7.9% (SD, 12.5%). The decrease from May to July 2020 was notable compared with the same period during the previous year (Supplementary Table 2, Supplementary Figure 7) The total numbers of completed clinical trials with results in 2020 decreased compared with the number in 2019 in Africa this decrease in current clinical trials will affect new treatments for at least several years to come It is important to note that this study showed regional trends in addition to global trends of clinical trials conducted during the COVID-19 pandemic. Interestingly, there was a negative correlation between the number of COVID-19 cases and clinical trial activity. Demographic effects have been reported (19, 20) and it is clear that the number of COVID-19 cases varies by region where the number of COVID-19 cases per 1 million population was <100 at the end of April 2020 the percentage decrease in the numbers of newly started clinical trials and reported results was modest compared with the decrease seen in Europe and the United States where the number of COVID-19 cases per 1 million population was more than 2000 Given that international clinical trials are now frequently performed if a region is unable to participate in a pivotal international clinical trial due to the COVID-19 epidemic this will cause a delay in new drug development in that region This could exacerbate the existing global inequalities It is suggested that the control of COVID-19 by each country/region is also important for the development of future treatments in that country/region It is unlikely that COVID-19 will disappear in the near future and therefore some medical resources will continue to be dedicated to COVID-19 even in the post-COVID era It is necessary to consider budget and staffing levels with a view to developing new treatments for diseases other than COVID-19 To evaluate the impact of the COVID-19 pandemic on overall clinical development we examined the number of newly started clinical trials all indicators showed a marked drop in quarters 2 and 3 of 2020 compared with the same period during the previous year indicating the magnitude of the impact of the first wave of COVID-19 on clinical development The decrease in the number of clinical trials started might reflect the increase in workload of medical staff and/or the financial situation of sponsors The decrease in the number of completed studies might reflect the effect of delayed patient recruitment and/or required visits and the decrease in the number of submitted applications might reflect the termination of studies before they are competed and/or the increased workload of the applicant company and/or the regulatory authority Many experts are involved from the initiation of clinical trials to the submission of new drug applications thus it can be seen that the COVID-19 pandemic affected various stages of clinical development because information about an approved drug and its approval date is made publicly available it will be possible to evaluate the number of new approvals in another year or so During the first wave of the COVID-19 pandemic, there was a significant decline in clinical trial activity and subsequent new drug applications. This decline will affect the development of new treatments in the future, and it is necessary to recognize this situation and continue to make efforts to perform clinical trials for diseases other than COVID-19, as well as for COVID-19. Utilizing the COVID-19 prediction (30) it will be possible to reduce the impact by allocating resources so that activities for non-COVID clinical trials will not decrease before the rapid increase of COVID-19 infections in the future It takes a lot of time and effort to launch and complete a clinical trial and to establish a new treatment therefore a long-term perspective is required Even in a situation in which the COVID-19 pandemic is not yet under control it is important to implement concrete strategies that minimize the impact on new or ongoing clinical trials based on our analysis of the first wave of the COVID-19 pandemic The original contributions presented in the study are included in the article/Supplementary Material further inquiries can be directed to the corresponding author/s performed the analysis and interpreted the data All authors reviewed and approved the final draft This work was partly supported by JSPS KAKENHI Grant No YA reports grants and personal fees from Chugai Pharmaceutical Co. grants and personal fees from Daiichi Sankyo Company The remaining author declares that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher The Supplementary Material for this article can be found online at: https://www.frontiersin.org/articles/10.3389/fmed.2021.812370/full#supplementary-material Google Scholar Preserving clinical trial integrity during the coronavirus pandemic PubMed Abstract | CrossRef Full Text | Google Scholar Obstacles and considerations related to clinical trial research during the COVID-19 pandemic Geographical distribution of COVID-19 cases and deaths worldwide PubMed Abstract | CrossRef Full Text | Google Scholar The European medicines agency's EU conditional marketing authorisations for COVID-19 vaccines A COVID-19 vaccine: big strides come with big challenges PubMed Abstract | CrossRef Full Text | Google Scholar 8. 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Available online at: https://wwwdrugscom/support/abouthtml (accessed January 19 Determination of the coefficient of correlation PubMed Abstract | CrossRef Full Text | Google Scholar Statistics corner: a guide to appropriate use of correlation coefficient in medical research PubMed Abstract | Google Scholar User's guide to correlation coefficients PubMed Abstract | CrossRef Full Text | Google Scholar Lessons learnt from easing COVID-19 restrictions: an analysis of countries and regions in Asia Pacific and Europe FDA approval and regulation of pharmaceuticals PubMed Abstract | CrossRef Full Text | Google Scholar COVID-19 case-fatality rate and demographic and socioeconomic influencers: worldwide spatial regression analysis based on country-level data The confounded crude case-fatality rates (CFR) for COVID-19 hide more than they reveal-a comparison of age-specific and age-adjusted CFRs between seven countries Early transmissibility assessment of the N501Y mutant strains of SARS-CoV-2 in the United Kingdom doi: 10.2807/1560-7917.ES.2020.26.1.2002106 Reporting clinical studies affected by the COVID-19 pandemic: guidelines for authors Association between administration of systemic corticosteroids and mortality among critically ill patients with COVID-19: a meta-analysis Association between administration of IL-6 antagonists and mortality among patients hospitalized for COVID-19: a meta-analysis FDA efficiency for approval process of COVID-19 therapeutics and emergency use authorizations during the COVID-19 pandemic Trial reporting in clinicaltrials.gov - the final rule Missing clinical trial data: the evidence gap in primary data for potential COVID-19 drugs Public disclosure of the filing of new drug and therapeutic biologics applications with the US food and drug administration COVID-19 cases prediction in multiple areas via shapelet learning PubMed Abstract | CrossRef Full Text | Google Scholar Citation: Nishiwaki S and Ando Y (2021) COVID-19 Pandemic and Trends in Clinical Trials: A Multi-Region and Global Perspective Received: 10 November 2021; Accepted: 08 December 2021; Published: 24 December 2021 Copyright © 2021 Nishiwaki and Ando. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY) distribution or reproduction in other forums is permitted provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited in accordance with accepted academic practice distribution or reproduction is permitted which does not comply with these terms *Correspondence: Satoshi Nishiwaki, bi0zMTA0QHRmNy5zby1uZXQubmUuanA= Disclaimer: All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher 94% of researchers rate our articles as excellent or goodLearn more about the work of our research integrity team to safeguard the quality of each article we publish Thirteen Renton School District students returned last Saturday from an experience “that will last a lifetime,” says one of the principals Monday (May 5) for more than 3,200 seats on city councils Development is inspired by future transit center I'm Bob Sherman and today we take one of our occasional trips to the Manhattan School of Music to meet some of the talented teenagers in two of the pre-college division's four orchestras; here on this 2,162nd edition of the McGraw Family's Young Artist Showcase I didn't misspeak earlier there are indeed four orchestras in MSM pre- college division nearly 300 youngsters spend their Saturdays taking private lessons and then rehearsing within one of those symphonic ensembles Our own musical adventures as always are presented with high community pride by the Harold W One of the very impressive elements of these pre-college orchestral programs that MSM but 20th-century works that would pose significant challenges (I suspect) for a professional orchestra Nathan Hetherington guiding his pre-college Philharmonic youngsters through the rhythmic tangles of David Diamond's Symphony Number One Bob Sherman No applause because the rest of David Diamond's first symphony yet to come in this May 11th concert by the 16 and 17 year old players in the Manhattan school's Pre-College Philharmonic conducted by Nathan Hetherington Nell Flanders led the even younger members of the Pre-College Symphony and it's similarly a daunting work that they (as I suspect most of us) had never even heard of before: William Grant Still's Afro-American symphony This historically significant work: -it was the first symphony by an African-American composer to be played by a major American orchestra (I think it was back in 1930 or 31) Bob Sherman It's in four movements each bearing lines from a poem by Paul Laurence Dunbar We don't have time for the complete symphony It's mighty tiresome lying around on this sorrow-laden earthly ground and oftentimes I think it would be a sweet thing just to die and go long home" Bob Sherman Paul Laurence Dunbar's poetic thoughts for the finale of William Grant Still symphony are upbeat and exultant "Be proud my race in mind and soul/thy name is written on glory scroll/in characters of fire/high amid the clouds of Fame's Bright Sky/the banner emblazoned folds now fly/and truth shall lift them higher." Bob Sherman Part of the slow movement and the finale of William Grant Still's Afro-American Symphony.. Nell Flanders leading the 14 and 15 year old players in the Manhattan School's Pre-College Division Symphony We'll rejoin the older kids in the Pre-College Philharmonic and meet the two winners of its' own Concerto Competition after we call a short intermission here on the McGraw Family's Young Artists Showcase Bob Sherman I'm Bob Sherman and as promised we return to the Manhattan School of Music Graduation day this past May 11th when all four of the Pre-College orchestras were in performing action The most advance of the ensembles: the Pre-College Philharmonic and the two top winners were featured that evening Abigail Nishiwaki; then a graduating senior at Scarsdale High School studying at MSM with Elizabeth Faidley; now a freshman at Princeton where she plans to divide her attention between music and public policy.. Abigail also served as concertmaster of the Pre-College Philharmonic and her concerto choice was the first movement of the Sibelius Bob Sherman Quite a compelling performance: first moment from the D minor Concerto of John Sibelius played last May by the winner of the Manhattan School's Pre-College Philharmonic Concerto Competition The Pianist laureate of the concerto competition was 16 year-old Esteban Castro quite a number of awards and prizes have been for his jazz performances he's also composed more than 40 pieces and looks with a special pride on a big band commission from the Jazz at Lincoln Center Orchestra Esteban Castro did stay within the  classical world to collect the Manhattan School prize though and here is his exciting performance of Prokofiev's Piano Concerto Number one again with music director Nathan Hetherington conducting Bob Sherman Resounding and well-deserved applause for that sterling account of Prokofiev's Piano Concerto Number one played just a couple of months ago by 16 year-old Esteban Castro with Nathan Hetherington at the helm of the other teenagers in the Manhattan School's Pre-College Philharmonic director of the Pre-College division for making these performances available to us And that does end our showcase adventures for tonight meaning that it's time for our weekly greeting from the fellow whose personal enthusiasm and financial generosity have combined to let the Young Artist Showcase move about halfway through it's 42nd consecutive year on WQXR There's nothing more satisfying than seeing some of the wonderful young musicians we introduce on young artist showcase go on to become part of our musical mainstream And it happens all the time on this program It shows we're not only on the right track but leading the way towards a great American future in the arts Last month Astral Artists announced the 2019 winners of it's national auditions but next week we'll be eavesdropping on recitals by some of the 2018 laureates; an occasion to which you are most cordially invited Our Young Artists Showcase far in-away the longest ongoing series in WQXR's history is supported as a community service by Terry McGraw and the family foundation named for his dad McGraw Jr.  Max Fine as my stalwart producing partner with thanks due as well to you: members of our faithful listening panel Young Artists Showcase is a WQXR weekly radio show that since 1978 has sought out and displayed the talents of young emerging artists WQXR’s Young Artists Showcase is supported by The Harold W HYOGO: A long-awaited Arabic pita bread bakery ‘The Babylon Bakery’ opened it’s doors on Friday in Nishiwaki “Many Middle Eastern and Mediterranean Muslims live in the neighboring cities of Kobe Osaka as well as lots of other parts of Japan,” the bakery owner Iraqi-born Haithem Humadi told Arab News Japan “I hope they will be able to experience pita bread which is just like the one in their hometown,” he said The project was established after seven years of planning with great care taken to produce an authentic taste He imported the Syrian-designed pita bread manufacturing machine from Turkey he is going to produce 300 kilograms a day but the machine has so much capacity of 5 tonnes of bread per day the foreign technicians who are capable of maintaining these machines cannot come to Japan at present due to COVID restrictions “The Babylon Bakery” will mainly deal with businesses such as halal shops and restaurants throughout Japan although individuals will be able to buy online Humadi commented: “We posted on Facebook and Twitter only last week and received more than 200 responses in five days from Tokyo Hiroshima and other regions not only the people from overseas residing in Japan Japanese version Japanese version Japanese version Copyright THE MAINICHI NEWSPAPERS. All rights reserved. Michiko Nishiwaki’s career spans bodybuilding, martial arts, and action cinema. Born on November 21, 1957, in Japan, she has challenged norms and inspired change in multiple fields. With her action movies, she was a key figure in the martial arts boom era of filmmaking. In the early 1980s, Michiko Nishiwaki became Japan’s first female bodybuilding champion. She held the title for three years. Her success came without using performance-enhancing drugs. At a time when muscular women faced criticism, Nishiwaki showed that strength and femininity could coexist. Her achievements led to a fitness boom in Japan. She opened three gyms and appeared frequently on television. Nishiwaki is trained in several martial arts. She holds a black belt in Gōjū-ryū karate and has studied Shotokan karate, wushu, and taekwondo. These skills became central to her film career. View this post on Instagram A post shared by LowKick MMA (@lowkickmma) Nishiwaki debuted in the 1985 film My Lucky Stars with Jackie Chan and Sammo Hung. She often played villains and tough female characters in action films. Her roles helped popularize the “girls with guns” subgenre such as In the Line of Duty III and Avenging Quartet highlighted her martial arts and stunt abilities She became a key figure in Hong Kong action cinema during the 1980s and 1990s She worked as a stuntwoman for stars like Lucy Liu in Charlie’s Angels and Kelly Hu in The Scorpion King Her stunts appeared in films like Kill Bill: Vol Nishiwaki broke barriers for women in fitness She inspired more women to pursue strength training and martial arts Her work brought Hong Kong action styles to global audiences Timothy Wheaton is a combat sports writer who covers MMA He has been an avid follower of these sports since 2005 Tim also works with a host of other media sites such as Calf Kick Sports Tim is the authority on kickboxing and an MMA journalist who has covered K-1 in Persona 3 Reload with all dialogue options Here is the Strength Social Link Guide for Yuko Nishiwaki in Persona 3 Reload This includes the answers for Social Link events with Yuko Nishiwaki Yuko is located in the 2F Classroom Hallway in Gekkoukan High School and is available on Wednesdays and Saturdays as long as it is a school day not be available the week leading up to exams To begin the Strength Social Link for Yuko you must first start the Chariot Social Link you will get the option to greet Yuko and invite her to walk home with you This will then start the Strength Social Link having a Persona of the Strength Arcana in your stock will increase each interaction by one point and will speed up the progress of the Social Link The protagonist will receive the Kid's Letter from Yuko Nishiwaki when the Strength Social Link reaches Rank 10 This decision to enter a romantic relationship with Yuko Nishiwaki happens when leveling up the Social Link to Rank 9 Here are all of the dialogue options and answers for Yuko Nishiwaki in Persona 3 Reload Dialogue options in bold are the best options a friendly reminder to have a Persona of the Strength Arcana when you spend time with Yuko Choosing “I'm honored” or “I don't mind” may be Romance flags for Yuko Choosing “Wanna make that true?” may be a Romance Flag for Yuko Only digital versions are available for Xbox Jesseyriche Cortez wrote about video games for ClutchPoints Jesseyriche also reviews games and plays the critically acclaimed MMORPG Final Fantasy XIV that has a free trial up to level 70 and includes the entirety of A Realm Reborn and Stormblood with no restrictions on playtime Here is our Strength Social Link guide for Yuko Nishiwaki in Persona 3 Portable The guide may contain spoilers, so do be careful. It is also important to note that this guide is for Persona 3 Portable, which originally came out in 2010, as well as in 2023 with the port. The Social Link Guide for Strength in Persona 3 Reload is a different guide altogether You can find Yuko during the day outside Classroom 2-F in Gekkoukan on Wednesdays and Saturdays the Protagonist does not get additional skills during battle If you choose to play as the male protagonist, you can start the Social Link as early as April 28. To start it, the player must ask to walk her home during Ranks 2 and 3 of the Chariot Social Link You can also romance Yuko as the male protagonist The affection points below assume that you have a Persona of the Strength arcana with you If you choose to play as the female protagonist, you will not have Yuko as your Strength Social Link. Instead, Koromaru will be your Strength Social Link That's all for our guide on Persona 3 Portable's Strength Social Link, Yuko Nishiwaki. Persona 3 Portable is available on PC. For more gaming news from us, you can check out our gaming news articles Yuko Nishiwaki is the team manager of the Track Team you signed up for You’ll have to jump through some slight hoops to unlock her Social Link This guide will be completely spoiler-free If you’re in the middle of Yuko’s social link this guide will only list the answers and avoid spoilers altogether Each Social Link allows you to interact with characters outside of S.E.E.S. allowing you to experience a fun or emotional side story in the meantime you’ll encounter some prompts where you’ll have to respond to a question with a specific answer Some of these answers will reward you with points that allow you to proceed through a Social Link faster We’ll list out the best answers for you to pick for each rank so you can run through each Social Link efficiently Important Note: You can obtain even more points by having a Persona with a matching Arcana in your party if you have a Strength Persona in your arsenal you’ll be able to go through Yuko’s Social Link even faster Yuko’s Social Link is easy to unlock but requires you to unlock the Chariot Social Link first you’ll meet Yuko and have the option to ask her to walk home with you but as you progress through the Chariot Social Link she will eventually agree to walk home with you no choices you make will impact the course of your Social Link as you will automatically reach the final rank no matter which choice you make Writer and journalist covering the gaming industry for The Nerd Stash