OTCQX:JAGGF) has announced significant growth in its mineral reserves and resources for 2024 The company reported a 63% increase in Proven and Probable Mineral Reserves to 764,000 ounces (5,903 kt @ 4.03 g/t Au) Key highlights include the first Mineral Reserves at Onças de Pitangui project of 284,000 ounces a 16% increase in Faina zone reserves to 160,000 ounces and stable Measured and Indicated Resources at 1,659,000 ounces The company's Inferred Mineral Resources grew to 1,676,000 ounces The Life of Mine plans show five years of production at Pilar starting in 2025 operations at the Turmalina mine are currently suspended due to an incident at the MTL Complex OTCQX:JAGGF) ha annunciato una crescita significativa delle sue riserve e risorse minerarie per il 2024 L'azienda ha riportato un aumento del 63% delle Riserve Minerarie Provate e Probabili a 764.000 once (5.903 kt @ 4,03 g/t Au) I principali punti salienti includono le prime Riserve Minerarie presso il progetto Onças de Pitangui di 284.000 once un aumento del 16% delle riserve nella zona Faina a 160.000 once e risorse misurate e indicate stabili a 1.659.000 once Le Risorse Minerarie Indeterminate dell'azienda sono cresciute a 1.676.000 once dimostrando un forte potenziale di crescita I piani di Vita della Miniera mostrano cinque anni di produzione a Pilar a partire dal 2025 prevede una produzione d'oro oltre il 2030 le operazioni presso la miniera Turmalina sono attualmente sospese a causa di un incidente nel Complesso MTL OTCQX:JAGGF) ha anunciado un crecimiento significativo en sus reservas y recursos minerales para 2024 La compañía reportó un aumento del 63% en las Reservas Minerales Probadas y Probables a 764,000 onzas (5,903 kt @ 4.03 g/t Au) Los aspectos más destacados incluyen las primeras Reservas Minerales en el proyecto Onças de Pitangui de 284,000 onzas un aumento del 16% en las reservas de la zona Faina a 160,000 onzas y recursos Medidos e Indicados estables en 1,659,000 onzas Los Recursos Minerales Inferidos de la compañía crecieron a 1,676,000 onzas demostrando un fuerte potencial de crecimiento Los planes de Vida de la Mina muestran cinco años de producción en Pilar comenzando en 2025 proyecta producción de oro más allá de 2030 las operaciones en la mina Turmalina están actualmente suspendidas debido a un incidente en el Complejo MTL OTCQX:JAGGF)은 2024년을 위한 광물 매장량 및 자원의 상당한 성장을 발표했습니다 회사는 검증된 및 추정된 광물 매장량이 63% 증가하여 764,000 온스(5,903 kt @ 4.03 g/t Au)로 주요 하이라이트는 온카스 드 피탕귀 프로젝트의 첫 광물 매장량이 284,000 온스에 이르며 회사의 추정 광물 자원은 1,676,000 온스로 증가하여 강력한 성장 잠재력을 보여줍니다 광산 생애 계획은 2025년부터 시작되는 필라르에서의 5년간의 생산 계획을 보여주며 온카스 드 피탕귀를 포함한 MTL 복합체는 2030년 이후에도 금 생산을 계획하고 있습니다 그러나 터말리나 광산의 운영은 현재 MTL 복합체에서 발생한 사건으로 인해 중단된 상태입니다 OTCQX:JAGGF) a annoncé une croissance significative de ses réserves et ressources minérales pour 2024 L'entreprise a rapporté une augmentation de 63 % des réserves minérales prouvées et probables à 764 000 onces (5 903 kt @ 4,03 g/t Au) Les points clés incluent les premières réserves minérales du projet Onças de Pitangui de 284 000 onces une augmentation de 16 % des réserves dans la zone Faina à 160 000 onces et des ressources mesurées et indiquées stables à 1 659 000 onces Les ressources minérales inférées de l'entreprise ont augmenté à 1 676 000 onces démontrant un fort potentiel de croissance Les plans de vie de la mine prévoient cinq ans de production à Pilar à partir de 2025 projette une production d'or au-delà de 2030 les opérations à la mine Turmalina sont actuellement suspendues en raison d'un incident survenu dans le complexe MTL OTCQX:JAGGF) hat ein signifikantes Wachstum seiner Mineralreserven und -ressourcen für 2024 bekannt gegeben Das Unternehmen berichtete von einem 63%igen Anstieg der nachgewiesenen und wahrscheinlichen Mineralreserven auf 764.000 Unzen (5.903 kt @ 4,03 g/t Au) Wesentliche Highlights sind die ersten Mineralreserven im Projekt Onças de Pitangui von 284.000 Unzen ein Anstieg der Reserven in der Faina-Zone um 16% auf 160.000 Unzen und stabile gemessene und angezeigte Ressourcen von 1.659.000 Unzen Die geschätzten Mineralressourcen des Unternehmens wuchsen auf 1.676.000 Unzen Die Lebensdauerpläne der Mine zeigen fünf Jahre Produktion in Pilar ab 2025 eine Goldproduktion über 2030 hinaus prognostiziert Die Betriebe in der Turmalina-Mine sind jedoch derzeit aufgrund eines Vorfalls im MTL-Komplex ausgesetzt Boosts Proven and Probable Mineral Reserves by 63% TORONTO, ON / ACCESS Newswire / March 31 ("Jaguar" or the "Company") (TSX:JAG)(OTCQX:JAGGF) is pleased to announce its annual Mineral Reserves and Mineral Resources (MRMR) statement for 2024 Proven and Probable Mineral Reserves (2P Mineral Reserves) increased by 63% to 764 koz (5,903 kt @ 4.03 g/t Au) The Onças de Pitangui projectreported the first Mineral Reserves of 284 koz (2,122 kt @ 4.16 g/t Au) Measured and Indicated (M&I) Resources decreased by 1% net of depletion to 1,659 koz (12,325 kt @ 4.19 g/t Au) Inferred Mineral Resources increased to 1,676 koz (14,621 kt @ 3.56 g/t Au) highlighting the Company's strong growth potential Onças de Pitangui project measured and indicated resources increased by 2% to 457 koz (3,547 kt @ 4.01 g/t Au) and inferred resources increased by 29% to 490 koz (4,184 kt @ 3.64 g/t Au) Faina zone continued its upward trajectory with a 16% increase in 2P Mineral Reserves to 160 koz (1,019 kt @ 4.87 g/t Au) Life of Mine (LOM) plans at Pilar demonstrate five years of production starting in 2025 with the inclusion of the Onças de Pitangui reserves and resources demonstrates gold production well past 2030 "This year's Mineral Reserve and Mineral Resource update further advances our organic growth strategy With the initial disclosure of probable Mineral Reserves at our Onças de Pitangui project While we have challenges to overcome following the incident at our MTL Complex which resulted in the suspension of operations at our Turmalina mine once we are able to return to work at this mine we see a direct road to increase the ounce production across the Company is the key to having new mining areas that can take our production to a new level." Proven and Probable Mineral Reserves and Measured Indicated and Inferred Mineral Resources figures are reported for the Company's two operating mining and production complexes and for the Paciência complex (currently on care and maintenance) in the Iron Quadrangle Gold Province The MTL complex (currently suspended) is comprised of the Turmalina mine (orebodies A the Onças de Pitangui project as well as the Zona Basal and Pontal deposits The Caeté complex is comprised of the Pilar mine (orebodies BA the Roça Grande mine (on care and maintenance) and the Córrego Brandão open-pit deposit The Paciência complex is comprised of the Santa Isabel and Margazão mines (both on care and maintenance) and the Bahú deposit 2024 MINERAL RESERVES AND MINERAL RESOURCES The Mineral Resources and Mineral Reserves reported herein and respective changes in inventory were consolidated based on estimates sourced from the technical reports listed in Table 3 CIM (2014) definitions were followed for Mineral Reserves estimates The location map in Figure 1 indicates the location of the Company's production complexes and respective mines and deposits The Mineral Rights are situated inside a 60 km radius from Belo Horizonte Schematic location map showing Jaguar's mineral rights and production complexes associated to the Rio das Velhas greenschists of the Iron Quadrangle Gold Province Bar chart of the consolidated 2024 Proven & Probable Mineral Reserves by operation The Company's Proven and Probable Mineral Reservesas at December 31 mainly due to the addition of 284 koz of Mineral Reserves from the Onças de Pitangui project (see bar chart in Figure 2) Detailed information of the Company's current consolidated Mineral Reserves inventory is shown in Table 1 at the end of this release A total of 572 koz of Mineral Reserves are attributed to the MTL complex Mineral Reserves distributed among the Turmalina mine Orebodies A the Faina zone and the Onças de Pitangui project 4 and 5 illustrate the spatial distribution of the Mineral Reserves and actual or planned development associated to the Turmalina mine The current Mineral Reserves inventory resulted from the following add-ons and depletions in 2024: Mineral Reserves were estimated by Jaguar staff reviewed and validated by SLR's Qualified Persons The current Proven and Probable Mineral Reserves of 128 koz (1,255 kt @ 3.26 g/t Au) for Orebodies A & C represent a net decrease of 16 koz from the Mineral Reserves established in December 2023 resulting from: The depletion of 29 koz resulting from mining activities in 2024; The subtraction of 5 koz resulting from a risk analysis performed in 2024 and The re-categorization of Mineral Resources into18 koz of Mineral Reserves resulting from in-fill / step-out drilling and geological model reinterpretation The Turmalina mine 2P Mineral Reserves are distributed among the following orebodies: The Faina zone hosts a total of 160 koz (1,019 kt @ 4.87 g/t Au) of Proven and Probable Mineral Reserves The Mineral Reserve addition of 27 koz attributed to the re-categorization of Mineral Resources exposed by the underground development and infill drilling that took place in 2024 The addition was offset by the production of 4 koz in 2024 resulting in a net increase of 22 koz compared to the prior year NI 43-101 Technical Report - Turmalina Mining Complex includes an update of Probable Mineral Reserves totalling 284 koz (2,122kt @ 4.16 g/t Au) at the Onças de Pitangui project Long Sections showing year-over-year changes in Turmalina mine Mineral Reserves 2024 vs 2023 Long Sections showing year-over-year changes of the Faina zone Mineral Reserves2024 vs 2023 Cross Section showing the Onças de Pitangui project Reserves as at December 31 Infill and step-out drilling and geological model reinterpretation executed in 2024 at the Pilar mine resulted in the addition of 43 koz to the Pilar mine mineral inventory This addition was offset by the extraction of 45 koz in 2024 the updated estimates of the Pilar mine Mineral Reserves total an amount of 192 koz (1536 kt @ 3.89 g/t Au) a 2 koz net decrease in the 2P Mineral Reserve categories if compared to the 194 koz (1,906 Kt @ 3.17 g/t Au) of 2P Mineral Reserves Mineral Reserves at the Pilar mine are divided between Orebody BA (38 koz The Longitudinal sections shown in Figure 6 illustrate the spatial distribution of the Mineral Reserves and actual development at the Pilar mine Long Section showing year-over-year changes in Pilar mine Mineral Reserves 2024 vs 2023 The Company's consolidated Mineral Resources as at December 31 include both the updated Mineral Resources for the Pilar and Turmalina mines as well as for the Onças de Pitangui project along with the unchanged Mineral Resources from previous disclosures for the Pontal and Zona Basal deposits and the Córrego Brandão and Bahu deposits Consolidated Measured and Indicated Mineral Resources decreased by 1% to 1,659 koz (12,325 kt @ 4.19 g/t Au) Inferred Mineral Resources increased by 3% to 1,676 koz (14,621 kt @ 3.56 g/t Au) which is a 48 koz net increase over the prior year Mineral Resources were estimated by Jaguar staff Detailed information of the Company's current consolidated Mineral Resources inventory as at December 23 is shown in Table 2 at the end of this release Graph showing the changes in M&I + I Resource Inventory in 2024 according to the estimation's updates and respective material sources Consolidated MTL complex (Measured and Indicated) Mineral Resources underground are reported as 1,104koz (8,219 kt @ 4.18 g/t Au) and Inferred Mineral Resources underground are reported as 906 koz (7,371 kt @ 3.82 g/t Au) Open pit Inferred Mineral Resources are reported as 32kozs (781 kt @ 1.28 g/t Au) The consolidated Mineral Resources are subdivided as follows: Turmalina mine Measured and Indicated Mineral Resources are reported as 360 koz Turmalina mine Inferred Mineral Resources are reported as 117 koz Faina zone Measured and Indicated Mineral Resources of 258 koz Faina zone Inferred Mineral Resources are reported as 193 koz Pontal deposit Measured and Indicated Mineral Resources are reported as 29 koz (266 kt @ 3.44 g/t Au) unchanged from 2023 disclosure Pontal deposit Inferred Mineral Resources are reported as 24 koz (159 kt @ 4.72 g/t Au) unchanged from 2023 disclosure Pontal South deposit Inferred Mineral Resources are reported as 81 koz (669 kt @ 3.76 g/t Au) unchanged from 2023 disclosure Onças de Pitangui project Measured and Indicated Mineral Resources of 457 koz Onças de Pitangui project Inferred Mineral Inferred Resources are reported as 490 koz The year-over-year changes in the Mineral Resources distribution at the Turmalina mine are illustrated in the longitudinal sections shown in Figures 8 Zona Basal deposit Inferred Mineral Resources (Open Pit) are reported as 32 koz unchanged since the initial estimates prepared by SLR reported in Technical Report on the Turmalina Complex Long Section of the Turmalina mine's Mineral Resources year-over-year changes (2024 vs Cross Section of the Onças de Pitangui project Mineral Resources year-over-year changes (2024 vs The Pilar mine was the only source of Mineral Resources inventory changes at the Caeté Complex The Mineral Resources estimates of the Pilar mine were prepared by Jaguar on site mine geology department under the coordination of the corporate geologists and mining engineers The database and procedures were revised by the SLR qualified persons The year-over-year changes in the Mineral Resources distribution at the Pilar mine are illustrated in the longitudinal section shown in Figure 11 Consolidated Caeté complex (Measured and Indicated) Mineral Resources Underground are reported as 556koz (4,106 kt @ 4.21 g/t Au) and Inferred Mineral Resources Underground are reported as 452 koz (3,597 kt @ 3.91 g/t Au) and Open Pit are reported as 51kozs (1,072 kt @ 1.48 g/t Au) subdivided as follows: Pilar mine Measured and Indicated Mineral Resources are reported as 435 koz Pilar mine Inferred Mineral Resources are reported as 335 koz Roça Grande mine Measure and Indicated Mineral Resources are reported as 121 koz Roça Grande mine Inferred Mineral Resources are reported as 117 koz Córrego Brandão deposit Inferred Mineral Resources are reported as 51 koz (1,072 kt @ 1.48 g/t Au) unchanged from 2021 disclosure Long Section showing year-over-year changes of the Pilar mine Mineral Resources 2024 vs The MTL Complex underground operations consist of several geological structures grouped into three orebodies - Orebodies A C (the Turmalina mine) - and the Faina zone where production began in 2024 through underground development responsible for the bulk of the Turmalina mine accumulated production estimated in 870 recovered koz with a strike length of approximately 250 m to 300 m Underground development has exposed approximately 1,100 vertical metres of Orebodies A and B mineralization along a robust down-plunge continuity lower grade lenses positioned in the foot-wall Orebody B mineralization has been outlined along a strike length of approximately 350 m to 400 m and to the same depths of Orebody A Orebody C is a series of northwest striking lenses generally of lower grade located to the southwest in the structural footwall of Orebody A Orebody C has replaced Orebody A as the Complex's principal production source Mineralization has been outlined along a strike length of approximately 800 m to 850 m B and C from Turmalina mine and the Pilar mine (Caeté Complex) share similar strong structural controls of the mineralization with the major gold mines known in the Iron Quadrangle Gold Province well known for their reduced strike length compared to very deep Explored in the past through diamond drilling the Faina zone is an orebody currently under development constituted by a series of offset striking parallel structures extending from surface to depth The focus of mining at Turmalina is shifting from Orebodies A and B to Orebody C and Faina zone Orebody C continues to grow with successful conversion of Resources to Reserves In addition to the development initiated at the Faina zone in 2024 the recent re-categorization of the Onças de Pitangui project Mineral Resources that resulted in 284 koz of Probable reserves will contribute to the MTL complex production growth The Pontal and Zona Basal deposits are unchanged from the previous year's estimates The Pilar Mineral Reserve 2024 inventory remained almost the same if compared to 2023 due to the gradual reduction in drilling activities and the depletion of 26 koz attributed to Orebody SW located in the opposed limb of the major structure that hosts the Pilar mine mineralization resulting from geological modelling parameters No changes of the Mineral Resources inventories were reported in 2025 in the Roça Grande mine and the Córrego Brandão deposit No changes in Mineral Resources inventories were reported in 2024 in the Santa Isabel and Marzagão mines Consolidated Mineral Reserves as at December 31 CIM (2014) definitions were followed for Mineral Reserves The released underground Mineral Reserves estimations include the Turmalina and Pilar mines the Faina zone Cut-off grades and constraints employed in the estimate such as minimum width and eventual minimum height for underground development and stopping are indicated in Table 4 The Exchange rate of R$5.20 / US$1.00 was applied to all the estimates Mineral Resources that are not Mineral Reserves do not have demonstrated economic viability Consolidated Mineral Resources as at December 31 CIM (2014) definitions were followed for the classification of Mineral Resources The released Mineral Resources estimations include the Turmalina and Onças de Pitangui project underground Mineral Resources and the Zona Basal deposit the Bahu and Córrego Brandão open-pit zones Mineral Resources Mineral Resources are inclusive of the Mineral Reserves at the Turmalina and Pilar mines the Faina zone and Onças de Pitangui project Drill hole and channel samples assay results freeze dates are indicated in Table 5 Freeze dates for depletion purposes relative to mining and forecast production volumes are shown in Table 5 Long-term gold price applied to the Mineral Resources estimations are informed in Table 5 Cut-off grades and constraints employed in the estimate such as minimum width and eventual minimum height for underground Mineral Resources and pit optimizations using Lerchs-Grossmann algorithm for open pit Mineral Resources estimations are indicated in Table 5 List of NI 43-101 Technical Reports and Internal Reports Employed to Support the Mineral Reserves and Resources Figures Development Parameters and Economic Premises Employed in the Mineral Reserves Estimates Development Parameters and Economic Premises Employed in the Mineral Resources Estimates INCIDENT AT THE MTL COMPLEX DRY-STACKED PILE a slump occurred in the north wall of the Satinoco dry-stack (waste rock and tailings) pile at the Company's MTL complex in the state of Minas Gerais approximately 130 kilometers northwest of the city of Belo Horizonte the Company's operating license at the complex's Turmalina mine was temporarily suspended until further notice The mine's personnel and nearby community members were evacuated before the slump occurred and there were no reported injuries or significant environmental impacts as a result of the incident For more information please see the press releases issued on December 9, 2024, December 30, 2024, January 7, 2025 and March 26, 2025, which can be found on the Company's website https://jaguarmining.com/investors/news-releases and under Jaguar's profile on SEDAR+ at https://www.sedarplus.ca The scientific and technical information contained in this press release has been reviewed and approved (i) in respect of the estimated Mineral Reserves and (ii) in respect of the estimated Mineral Resources (Pontal deposit) of SLR Consulting Canada Ltd (SLR) which office is situated at 55 University Avenue Pressacco are each "qualified persons" within the definition of NI 43-101 During the preparation of the updated Mineral Resource and Mineral Reserve estimates discussions were held on a weekly basis with Jaguar personnel and Deswik Brazil Holdings Pty Ltd Caeté and Paciência complexes several times Sepp's latest visit to the properties occurred in December 2022 All remaining scientific and technical information (other than described above) contained in this press release has been reviewed and approved by Jean-Marc Lopez BSc who is a "qualified person" as defined by NI 43-101 All sampling and samples utilized at Jaguar for Mineral Resource and or Mineral Reserves estimation uses a quality-control program that includes insertion of blanks and commercial standards in order to ensure best practice in sampling and analysis and BQ size drill core is sawn in half with a diamond saw Samples are selected for analysis in standard intervals according to geological characteristics such as lithology and hydrothermal alteration Rock channel sampling of the underground development follows the same standard intervals as for the drill core Half of the sawed sample is forwarded to the analytical laboratory for analysis while the remaining half of the core is stored in a secure location The drill core and rock chip samples for resource-reserve conversion and grade control samples are transported for physical preparation and analysis in securely sealed bags to the Jaguar in-house laboratory located at the company´s Caeté Complex Growth exploration samples are sent to the independent ALS Brazil (subsidiary of ALS Global) laboratory located in Vespasiano The analysis of these exploration samples is conducted at ALS Global's respective facilities (fire assay is conducted by ALS Global in Lima and multi-elementary analysis is conducted by ALS Global in Vancouver ALS has accreditation in a global management system that meets all requirements of international standards ISO/IEC 17025:2005 and ISO 9001:2015 All major ALS geochemistry analytical laboratories are accredited to ISO/IEC 17025:2005 for specific analytical procedures For a complete description of Jaguar's sample preparation, analytical methods, and QA/QC procedures, please refer to "Technical Report on the Roça Grande and Pilar Operations, Minas Gerais State, Brazil", a copy of which is available on the Company's SEDAR profile at www.sedarplus.ca Mineralized material for each orebody was classified into the Measured or Inferred Mineral Resource categories based on the search ellipse ranges obtained from the variography study the observed continuity of the mineralization and previous production experience from these orebodies The Mineral Resources are inclusive of Mineral Reserves For those portions of the Mineral Resources that comprise the Mineral Reserve stope design wireframes were used to constrain the outlines of the Mineral Resource volumes The Iron Quadrangle has been an area of mineral exploration dating back to the 16th century The discovery in 1699-1701 of gold contaminated with iron and platinum-group metals in the south-eastern corner of the Iron Quadrangle gave rise to the name of the town Ouro Preto (Black Gold) The Iron Quadrangle contains world-class multi-million-ounce gold deposits such as Morro Velho Jaguar Mining is the second largest operating gold company tenement holder in the Iron Quadrangle holding or having access to approximately 46,600 hectares distributed among 56 mining and exploration titles (23,800 hectares held by Jaguar and 22,900 hectares held by Iamgold) Vernon Baker Chief Executive Officer Jaguar Mining Inc. vernon.baker@jaguarmining.com416-847-1854 Alfred Colas Chief Financial Officer Jaguar Mining Inc. alfred.colas@jaguarmining.com416-847-1854 Certain statements in this news release constitute "forward-looking information" within the meaning of applicable Canadian securities legislation Forward-looking statements and information are provided for the purpose of providing information about management's expectations and plans relating to the future All of the forward-looking information made in this news release is qualified by the cautionary statements below and those made in our other filings with the securities regulators in Canada Forward-looking information contained in forward-looking statements can be identified by the use of words such as "are expected," "is forecast," "is targeted," "approximately," "plans," "anticipates," "projects," "anticipates," "continue," "estimate," "believe" or variations of such words and phrases or statements that certain actions events or results "may," "could," "would," "might," or "will" be taken may be considered to be or include forward-looking information This news release contains forward-looking information regarding the duration of the indefinite suspension of the Company's MTL complex in the wake of the slump at its Satinoco dry tailings pile the Company's assessment of the environmental impact of the Satinoco tailings slump the expected operational impact of the tailings pile slump including the cost and timeline for (and feasibility of) recommencing operations at the Turmalina mine the future stability of the tailings pile in question and safety of the Turmalina mine the Company's assessment of the financial impact of legal claims regulatory fines and investigations related to the tailings pile slump including the likelihood of successful appeals or settlements which are currently ongoing and subject to a wide range of possible outcomes the potential for new regulatory requirements operational restrictions and increased inspections imposed by Brazilian mining authorities the Company's ability to effectively manage relationships with affected community members repair its social license and mitigate reputational damage the potential for unforeseen environmental or human health consequences resulting from the tailings pile slump and the Company's ability to address any such issues the Company's expectations regarding its ability to secure sufficient financing and maintain liquidity in light of the financial burdens associated with the tailings pile slump the Company's plans for stakeholder engagement risk mitigation and corporate responsibility initiatives aimed at ensuring long-term sustainability management's expectations regarding the Company's response to the tailings pile slump and the Company's recovery and remediation efforts at the MTL complex any information and statements related to expected growth the timing and amount of estimated future production costs and timing of the development of projects and new deposits restarting suspended or disrupted operations The Company has made numerous assumptions with respect to forward-looking information contained herein and the level and volatility of the price of gold; the accuracy of reserve and resource estimates and the assumptions on which the reserve and resource estimates are based; the receipt of necessary permits; market competition; ongoing relations with employees and impacted communities; political and legal developments in any jurisdiction in which the Company operates being consistent with its current expectations including the impact of any potential power rationing exploration and mine operating licenses and permits being obtained and renewed and/or there being adverse amendments to mining or other laws in Brazil and any changes to general business and economic conditions Forward-looking information involves a number of known and unknown risks and uncertainties including among others: the risk of Jaguar not meeting the forecast plans regarding its operations and financial performance; uncertainties with respect to the price of gold environmental compliance and change in environmental legislation and regulation weather delays and increased costs or production delays due to natural disasters procurement and delivery of parts and supplies to the operations; uncertainties inherent to capital markets in general (including the sometimes volatile valuation of securities and an uncertain ability to raise new capital) and other risks inherent to the gold exploration may cause actual results to differ materially from those anticipated by the Company and described herein there are risks and hazards associated with the business of gold exploration industrial accidents and workplace safety problems unusual or unexpected geological formations procurement fraud and gold bullion thefts and losses (and the risk of inadequate insurance readers should not place undue reliance on forward-looking information For additional information with respect to these and other factors and assumptions underlying the forward-looking information made in this news release see the Company's most recent Annual Information Form and Management's Discussion and Analysis as well as other public disclosure documents that can be accessed under the issuer profile of "Jaguar Mining Inc." on SEDAR+ at www.sedarplus.ca The forward-looking information set forth herein reflects the Company's reasonable expectations as at the date of this news release and is subject to change after such date The Company disclaims any intention or obligation to update or revise any forward-looking information The forward-looking information contained in this news release is expressly qualified by this cautionary statement Already have an account? Login Volume 6 - 2015 | https://doi.org/10.3389/fmicb.2015.01526 This article is part of the Research TopicRecent advances in the study of the host-fungus interactionView all 13 articles Histoplasma capsulatum is responsible for a human systemic mycosis that primarily affects lung tissue Macrophages are the major effector cells in humans that respond to the fungus and the development of respiratory disease depends on the ability of Histoplasma yeast cells to survive and replicate within alveolar macrophages capsulatum is a decisive step in the yeast dissemination into host tissues Although the role played by components of cell-mediated immunity in the host's defense system and the mechanisms used by the pathogen to evade the host immune response are well understood knowledge regarding the effects induced by H capsulatum in host cells at the nuclear level is limited capsulatum yeast cells display a unique architectural arrangement during the intracellular infection of cultured murine alveolar macrophages characterized as a formation of aggregates that seem to surround the host cell nucleus resembling a “crown.” This extranuclear organization of yeast-aggregates generates damage on the nucleus of the host cell producing DNA fragmentation and inducing apoptosis even though the yeast cells are not located inside the nucleus and do not trigger changes in nuclear proteins The current study highlights a singular intracellular arrangement of H capsulatum yeast near to the nucleus of infected murine alveolar macrophages that may contribute to the yeast's persistence under intracellular conditions since this fungal pathogen may display different strategies to prevent elimination by the host's phagocytic mechanisms According to Newman et al. (2011) the destruction of alveolar macrophages and their subsequent ingestion by other immune cells are events that promote the propagation of the infection to different organs during the acute stage of primary histoplasmosis it is clear that the interaction between the macrophage and H capsulatum is a decisive step in the occurrence of yeast dissemination into host tissues Nuclear fragmentation is a morphological cellular alteration associated with apoptosis (Deepe and Buesing, 2012); thus, nuclear damage in host cells can be characterized as a cellular effect that contributes to the pathogenesis of histoplasmosis. Glukhov et al. (2008) reported that bacterial endotoxins induce nuclear DNA damage in human mononuclear cells which is associated with the infectious process and disease manifestation knowledge of the DNA fragmentation induced by microorganisms is limited it is necessary to investigate the behavior of nuclear envelope proteins during infection The labeling of nuclear envelope proteins in host cells could contribute to the characterization of the behavior of these proteins during the course of an in vitro infection capsulatum interacts with macrophages and evades host immune defenses have been well documented this is the first report that attempts to characterize the interaction pattern and the nuclear damage of parasitized host cells after the internalization of H capsulatum yeast in order to verify the correlation of these yeast cells with host cell integrity Apoptosis assays were performed as well as the staining of nuclear envelope proteins in host cells infected with the fungus Our study highlights the intracellular behavior and the effects induced by H capsulatum at a nuclear level in cultured infected alveolar macrophages The 60I strain was isolated from a human clinical case and was deposited in the collection of the Clinical Mycology Laboratory of the Faculty of Pharmaceutical Sciences Yeasts were grown in brain–heart infusion (BHI-broth) (Difco Laboratories USA) and supplemented with 0.1% L-cysteine and 1% glucose capsulatum yeast cells were washed three times with phosphate-buffered saline (PBS) followed by low-speed centrifugation for 1 min at 600 × g to remove large yeast clumps Suspensions of single yeast cells were separated for counting with a hematocytometer were cultured overnight at 37°C on coverslips placed in the well-bottom of 24-well plates (TPP® Switzerland) using Dulbecco's modified Eagle's medium (DMEM) (Sigma-Aldrich USA) supplemented with 10% heat-inactivated fetal calf serum (Cultilab They were processed exactly as outlined in the experimental protocol recommended by the Ethics Committee on Animal Experiments of the Faculty of Pharmaceutical Sciences of Araraquara—UNESP (reference number: 10/2011/CEUA/FCF) All efforts were made to minimize suffering in all animal procedures H. capsulatum cell-free antigen, a rich solution of cell wall associated antigens, was prepared as described previously by Sá-Nunes et al. (2005) Protein concentration was quantified using the Bradford method (BioRad Laboratories Inc. To prepare a polyclonal antibody raised against cell-free antigen of H rabbits were inoculated by intradermal injection of 1.0 mL of the cell-free antigen mixed with 1.0 mL of complete Freund's adjuvant Subsequent injections of this antigen with incomplete Freund's adjuvant were given weekly for a period of 4 weeks The rabbits were bled at the 7th day after the last dose The immunoglobulin fraction of each rabbit anti-serum was separated by precipitation with ammonium sulfate and stored at −70°C For this assay, a reference strain from the American Type Culture Collection (ATCC), G-217B, was compared with strains EH-315 and 60I. The infection rate of each strain was estimated using the AMJ2-C11 alveolar macrophage cell-line (ATCC, CRL-2456). The assay was performed in 24-well plates (TPP®) containing 105 AMJ2-C11 macrophages per well, as described by Sardi et al. (2012) Each cultured macrophage monolayer was infected with 500 μL of yeast inoculum (1 × 106 yeasts/mL) and plates were incubated at 37°C for 0 a macrophage monolayer was washed three times with sterile PBS to remove released yeast cells the AMJ2-C11 cells were detached at 37°C for 2 min using trypsin-EDTA (Gibco Life Technologies 100 μL of each infected macrophage suspension was plated on supplemented BHI-agar (Difco) and incubated at 37°C fungal colonies were counted and the CFU/mL was estimated for each strain tested capsulatum yeast cells that was able to infect the alveolar macrophage monolayer at each incubation time a control for yeast cell viability was performed in which yeast cells were maintained with trypsin-EDTA for 2 min and Trypan blue solution was added afterward to detect viability capsulatum infection rate curves were constructed based on the data of each strain incubated at the different times Tests were set up in triplicate in two independent assays The interaction between alveolar macrophages and H capsulatum yeast were also monitored by conventional Giemsa staining and indirect fluorescence Samples of infected macrophages were maintained under the optimal culture conditions for a 5 h incubation The infected monolayers were fixed with 4% paraformaldehyde and permeabilized in 0.5% Triton X-100 for 30 min capsulatum antibody was added for a 1 h incubation at room temperature and unbound antibodies were removed by washing with PBS Alexa Fluor®594-conjugate goat anti-rabbit IgG (Invitrogen-Molecular Probes USA) was added and incubated for 1 h at room temperature and fluorescein isothiocyanate (FITC)-labeled phalloidin (Sigma-Aldrich USA) was added with 1 h of incubation at 37°C All nuclei were stained using 4′,6-Diamino-2-phenylindole (DAPI) (Sigma-Aldrich The infected and non-infected macrophages were washed three times with PBS and analyzed under fluorescence microscopy All the images were acquired by the IN Cell Analyzer 2000 System (GE Healthcare Bio-Sciences Corp. the percentage of the infected macrophage population and the number of yeast cells per macrophage were determined using Investigator IN Cell 1000 Workstation software (GE Healthcare Bio-Sciences Corp.) This software includes an accurate analysis module that allows reaching reliable results to measure the morphology and the fluorescence intensity of user-defined nuclear and cytoplasmic compartments cells can be classified into subpopulations by applying one or more filters according to one or two user-selectable fluorescence or morphological events the AMJ2-C11 alveolar macrophages were counted as cells based on some parameters To measure yeast cells they were assumed as being “organelles,” and the following parameters were considered The final results were automatically obtained in a worksheet detailing the measures by well regarding the indicated parameters as numerical values AMJ2-C11 macrophage monolayers containing 105 macrophages per well were formed in 24-well plates (TPP®) 500 μL (1 × 106 yeasts/mL) of each inoculum of H capsulatum was stained with 10 μM carboxyfluorescein diacetate succinimidyl ester (CFSE) (Invitrogen capsulatum strains were added to their respective macrophage monolayer and the plates were incubated at 37°C the monolayers were washed three times with sterile PBS and macrophages were detached at 37°C for 2 min using trypsin-EDTA (Gibco Life Technologies) diluted in PBS Macrophage suspensions were harvested in Eppendorf tubes and centrifuged at 600 × g Supernatants were removed and PBS was added to each Eppendorf tube before cell counting by flow cytometry (BD FACSCanto Becton Dickinson we considered parameters related to the size (size forward scatter—FSC) granularity (granularity side scatter—SSC) and fluorescence of 10,000 cells per tube The results were determined through the fluorescence intensity (FI) of yeast cells labeled with CFSE as estimated by BD FACSDiva software Gates of specific population were viewed and analyzed by dot-plot These data allowed one to determine the percentage of infected alveolar macrophages and discriminate the infectivity of different strains of H Non-infected AMJ2-C11 alveolar macrophages and unlabeled yeast were used as negative controls in the assay Assays were performed in three biological replicates and two technical replicates AMJ2-C11 macrophages, in 24-well plates, were infected with H. capsulatum strains EH-315 or 60I using a standardized suspension of 1 × 106 yeasts/mL and incubated at 37°C for 5 h. Non-infected macrophages were used as a negative control. The alkaline version of the comet assay (single cell gel electrophoresis) was performed as described by Singh et al. (1988) Duplicate slides were prepared and stained with ethidium bromide We screened 50 AMJ2-C11 macrophages per sample with a fluorescence microscope (Carl Zeiss GmbH Germany) equipped with a 515–560 nm excitation filter The level of DNA damage was assessed by an image analysis system (TriTek CometScore and the DNA percentage in comet tail was obtained for each treatment the percentage of the macrophage population that showed DNA damage was determined DNA fragmentation in infected macrophages was evaluated using TUNEL (terminal deoxynucleotidyl transferase dUTP nick-end labeling) staining following the protocol recommended by the manufacturer (Roche Diagnostics which has the feature of specific labeling of fragmented DNA sequences that occur during the process of apoptosis capsulatum strains EH-315 and 60I in AMJ2-C11 macrophages cultured in 96-well plates The macrophages were incubated with a standardized suspension (1 × 106 yeasts/mL) of H capsulatum EH-315 or 60I and infection was allowed for 30 min Non-infected macrophages were used as a negative control macrophages were PBS washed and fixed in 4% paraformaldehyde for 1 h at room temperature Samples were washed three times with cold PBS and incubated with 200 μL permeabilization solution (0.05 M Tris and 2.5 mg/mL proteinase K) for 15 min at room temperature free reactive sites of the macrophage monolayer on the coverslips were blocked with 200 μL of a solution containing 3% bovine serum albumin and 20% fetal bovine serum in PBS at 37°C for 1 h the monolayers were washed three times with cold PBS and incubated with the components of the “TUNEL” mixture (dUTP solution containing the enzyme FITC-conjugated and “terminal deoxynucleotidyl transferase”) at 37°C the 3′ ends of the apoptotic DNA fragments were incorporated into the FITC-labeled nucleotides This reaction was catalyzed by terminal transferase and 100 μL of 1% paraformaldehyde was added per well Analysis of DNA fragmentation in macrophages was conducted to compare the EH-315 and 60I strains using non-infected macrophages as a negative control Images were captured using the IN Cell Analyzer 2000 System for light microscopy and were analyzed by Investigator IN Cell 1000 Workstation software (GE Healthcare Bio-Sciences Corp.) The results were evaluated using as parameter the fluorescence intensity emitted by the nucleus in each condition tested were infected with a standardized suspension of 1 × 106 yeasts/mL of H capsulatum strains EH-315 or 60I at 37°C for 5 h Nuclear envelope proteins were marked in either infected or non-infected AMJ2-C11 macrophages (negative control) infected and non-infected macrophage samples were fixed with 4% paraformaldehyde washed with PBS and permeabilized with 0.5% Triton X-100 for 30 min blocking was performed with 2.5% bovine serum albumin Unbound antibodies were removed by washing in PBS Alexa Fluor®594-conjugate goat anti-rabbit IgG (secondary antibody) was added for 1 h anti-SUN2 antibody (Santa Cruz Biotechnology Inc. or anti-Emerin antibody (Abcam) obtained in mice was added as a primary antibody and macrophage samples were incubated overnight Unbound antibodies were removed by PBS washing and a secondary Alexa Fluor®488-conjugate goat anti-mouse IgG antibody was added for 1 h The infected and non-infected macrophages were then washed three times with PBS and analyzed under confocal laser scanning microscopy (Leica TCS SP5 Confocal Microscopy System) Data were analyzed using Origin 7.0 (Origin Lab ANOVA was used to compare groups in CFU and TUNEL assays with the Bonferroni post-test P was calculated by Student's t-test and P ≤ 0.001 were considered statistically significant the infected and non-infected macrophages (control) were compared using the Kruskal–Wallis test and the associated Dunn post-test with P ≤ 0.05 were considered as statistically significant The results suggested that at 120 min of contact between alveolar macrophages and strain EH-315 the number of yeast cells in the macrophages increased the number of yeast cells was increased at 60 min and at 120–180 min the infection rate of strain 60I increased again at 300 min Infection rate assays of AMJ2-C11 alveolar macrophage with H capsulatum yeasts were considered using CFU/mL and statistics were performed by Two-way ANOVA with the Bonferroni post-test 60I strain and #P < 0.001 for EH-315 vs Infection of macrophages by H. capsulatum yeast (strains EH-315 and 60I) was also evaluated using Giemsa staining. This methodology was very useful for analyzing how yeasts interact with host macrophages (Figure 2) this staining did not provide accurate localization of yeast cells within phagocytes Giemsa staining of AMJ2-C11 macrophages after 5 h of H (A,B) Control of non-infected macrophages incubated with PBS The results are representative of two assays Indirect fluorescence microscopy, using images of the IN Cell Analyzer, revealed several intracellular yeasts in infected macrophages. An interesting finding was detected with this methodology, where intracellular H. capsulatum yeast cells aggregated in an architectural shape apparently surrounding the macrophage nucleus, resembling a “crown” (Figure 3) additional analyses with Investigator IN Cell 1000 Workstation software showed that strain EH-315 infected 95% of the macrophage population with an infection multiplicity up to 30 cells per macrophage whereas strain 60I infected 86% of macrophages with up to 24 yeast cells per macrophage capsulatum yeast cells on the infection of AMJ2-C11 macrophages capsulatum yeast cells were incubated at 37°C for 5 h (see details in Section Materials and Methods) (A,B) AMJ2-C11 macrophages infected with the EH-315; (C) AMJ2-C11 macrophages infected with 60I; and (D) non-infected AMJ2-C11 macrophages Indirect immunofluorescence: FITC-phalloidin in green showing the macrophage cytoplasm; Alexa Fluor®594 in red to yellow staining H capsulatum yeast cells; DAPI in blue staining macrophages nucleus Images were obtained using IN Cell Analyzer light microscopy Arrows indicate the architectural arrangement of yeast-aggregates apparently surrounding the nuclei of the phagocytic cells Results were expressed as FI of yeast labeled with CFSE and correspond to the fluorescence data of 10,000 cells per tube. To quantify the percentage of yeast cells bound to or within AMJ2-C11 macrophages, combination of two gates were applied to yeast cells and AMJ2-C11 cell-line (Figure 4) immediately the percentage of yeast cells interacting with AMJ2-C11 alveolar macrophages was determined capsulatum infection in alveolar macrophages strains EH-315 and 60I showed high infection rates in AMJ2–C11 macrophages both strains have a similar potential for infection because they are able to infect murine alveolar macrophages at rates of 98.34 and 96.52% The results represent the average of three independent assays set up in triplicate Dot-plot and histogram profiles of AMJ2-C11 alveolar macrophages and CFSE-labeled H (A,B) Population of labeled yeast cells (EH-315 and 60I strain respectively); (C) specific gate for AMJ2-C11 alveolar macrophages; and (D,E) Infected alveolar macrophages by EH-315 and 60I strain The cells were analyzed by flow cytometer BD FACSCanto Comet assay data of AMJ2-C11 macrophages infected with H (A) Images were acquired using a fluorescence microscope equipped with a 515–560 nm excitation filter and a 590 nm barrier filter (B) Evaluation of nuclear fragmentation in AMJ2-C11 macrophages infected with H Values are representative of the percentage of fragmented DNA present in the comet's tail (C) Percentage of macrophage population showing DNA damage in infected and non-infected (control) cells Data were analyzed by Kruskal-Wallis test with Dunn's post-test strain EH-315 was compared; #P < 0.05 when control vs TUNEL staining of alveolar macrophages after H (A) DNA fragmentation in the nucleus of AMJ2-C11 macrophages infected with H and 5 h after infection of alveolar macrophages with H Infected alveolar macrophages were compared with non-infected macrophages incubated with PBS (control) Each photomicrograph was processed by IN Cell Analyzer light microscopy using a 20X power field (B) Detection of nuclear fluorescence intensity derived from DNA fragmentation in AMJ2-C11 macrophages infected with H capsulatum strain EH-315 or 60I and non-infected macrophages (control) TUNEL-positive macrophages are represented by the nuclear fluorescence intensity with values derived from analysis of images using Investigator IN Cell 1000 Workstation software Scores given are the mean ± S.D and statistics were performed by Two-way ANOVA with the Bonferroni post-test These events were not found in non-infected macrophages Labeling of nuclear envelope proteins SUN2 In Panels (A–C) show confocal scanning microscopy images of representative AMJ2-C11 macrophages infected with H capsulatum strains EH-315 and 60I at 37°C for 5 h Non-infected macrophages were processed as a control The arrows show phagosome formation where several yeast cells are present Alexa Fluor®488 and Alexa Fluor®594 conjugates were used as secondary antibodies to reveal nuclear proteins (green) and yeast cells (red) Bright field images were merged with Alexa Fluor®594 and DAPI stain with orthogonal z stack sections after 5 h of macrophage-yeast infection Interactions of pathogenic fungi with host tissues are essential factors in the pathogenesis of mycoses (Tronchin et al., 2008) The present study demonstrated particular characteristics of the interaction between H capsulatum yeast cells and cultured murine alveolar macrophages capsulatum strains isolated from different sources based on their behavior and potential infection for AMJ2-C11 alveolar macrophages The EH-315 strain developed higher virulence (LD50 3 × 105 yeasts/mL) when compared to the 60I strain (LD50 3 × 108 yeasts/mL) under experimental conditions using an LD50 assay in male BALB/c mice (ML Taylor several factors contribute to non-optimize microorganisms plating that routinely reaches only 30% effectiveness for H as the CFU number is generally lower than the number of viable yeasts plated as strain G-217B showed delayed infection potential against host cells capsulatum strains on alveolar macrophages over a period of 5 h was characterized by a variable behavior (increases and decreases) in the yeast infection rate of alveolar macrophages We hypothesize that this profile occurs as a result of the dynamic interactions between yeast and macrophage membrane receptors H. capsulatum is a pathogen that commonly survives within macrophages by developing several intracellular evasion mechanisms (Strasser et al., 1999; Sebghati et al., 2000) Microscopic images obtained by indirect immunofluorescence assays showed a singular pattern of H capsulatum in the infected macrophages under in vitro conditions which was similar for the two fungal strains tested the yeast cells of both strains were able to form aggregates in the cytoplasm of the infected macrophages with an apparent distribution surrounding the macrophage nucleus after 5 h of infection These findings could be related to a new strategy for fungal intracellular survival Based on the potential for infection displayed by both strains of H. capsulatum in alveolar macrophages, it became necessary to evaluate the genotoxic potential of this fungus in AMJ2-C11 macrophages to identify the ability of H. capsulatum to induce damage to DNA in host cells. According to Yang et al. (2011) genotoxic agents chemically interact with the genetic material and cause oxidative changes or disruptions in the DNA molecule apoptosis allows the host to develop an effective response against infectious diseases such as tuberculosis it was possible to observe diffuse distribution of these proteins outside on the nuclear envelope in both infected and non-infected macrophages it is difficult to determine whether other isoforms are also present due to the absence of specific antibodies capsulatum yeast-aggregates were able to cause damage in the nuclear DNA and induce apoptosis in alveolar macrophages after 5 h of infection This damage to the DNA of macrophages while the yeast cells are not located inside the core could be a fungus strategy for the facilitation of its persistence throughout the host infection This finding has never been previously described the intracellular arrangement and the occurrence of effects induced by H capsulatum yeast-aggregates during the infection could promote the survival of the pathogen in the hostile conditions of the intracellular environment while also contributing to host tissue damage CS and AF conceived and designed the study RS and FS performed the experiments and analyzed the data FS and CS collaborated with reagents/materials/analysis tools All authors read and approved the final manuscript NS and AF wrote the paper with contributions from GR The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest decision to publish or manuscript preparation The Supplementary Material for this article can be found online at: http://journal.frontiersin.org/article/10.3389/fmicb.2015.01526 Apoptosis modulates protective immunity to the pathogenic fungus Histoplasma capsulatum Rapid quantification of pathogenic fungi by Cellometer image-based cytometry Flow cytometric quantitation of yeast a novel technique for use in animal model work and in vitro immunologic assays Coupling of the nucleus and cytoplasm: role of the LINC complex Apoptosis of Th1-like cells in experimental tuberculosis (TB) Deciphering the pathways of death of Histoplasma capsulatum-infected macrophages: implications for the immunopathogenesis of early infection Induction of apoptosis in A549 pulmonary cells by two Paracoccidioides brasiliensis samples Apoptosis of phagocytic cells induced by Candida albicans and production of IL-10 DNA damage in human mononuclear cells induced by bacterial endotoxin Mammalian SUN protein interaction networks at the inner nuclear membrane and their role in laminopathy disease processes The Histoplasma capsulatum vacuolar ATPase is required for iron homeostasis intracellular replication in macrophages and virulence in a murine model of histoplasmosis Lamin A/C and emerin regulate MKL1-SRF activity by modulating actin dynamics Immunization with apoptotic phagocytes containing Histoplasma capsulatum activates functional CD8(+) T cells to protect against histoplasmosis Emerin is hyperphosphorylated and redistributed in herpes simplex virus type 1-infected cells in a manner dependent on both UL34 and US3 Dendritic cells cross-present exogenous fungal antigens to stimulate a protective CD8 T cell response in infection by Histoplasma capsulatum Mechanical regulation of nuclear structure and function Histoplasma capsulatum inhibits apoptosis and Mac-1 expression in leucocytes Nesprin isoforms: are they inside or outside the nucleus PubMed Abstract | CrossRef Full Text Dendritic cells restrict the transformation of Histoplasma capsulatum conidia into yeasts Dynamics and molecular interactions of linker of nucleoskeleton and cytoskeleton (LINC) complex proteins Adhesion of Histoplasma capsulatum to pneumocytes and biofilm formation on an abiotic surface switch isoforms at the nuclear envelope during muscle development Sá-Nunes Efficacy of cell-free antigens in evaluating cell immunity and inducing protection in a murine model of histoplasmosis Profile of cytokines in the lungs of BALB/c mice after intra-nasal infection with Histoplasma capsulatum mycelial propagules Adhesion and invasion of Candida albicans from periodontal pockets of patients with chronic periodontitis and diabetes to gingival human fibroblasts Intracellular parasitism by Histoplasma capsulatum: fungal virulence and calcium dependence Sepúlveda Comparison of phylogenetically distinct Histoplasma strains reveals evolutionarily divergent virulence strategies A simple technique for quantitation of low levels of DNA damage in individual cells Regulation of the macrophage vacuolar ATPase and phagosome-lysosome fusion by Histoplasma capsulatum Membrane microdomain components of Histoplasma capsulatum yeast forms and their role in alveolar macrophage infectivity LINC complex alterations in DMD and EDMD/CMT fibroblasts and comparative genomic analysis of an invasive and non-invasive strain of Candida albicans Adherence mechanisms in human pathogenic fungi Capsular polysaccharides galactoxylomannan and glucuronoxylomannan from Cryptococcus neoformans induce macrophage apoptosis mediated by Fas ligand Nitric oxide synthase expression in macrophages of Histoplasma capsulatum-infected mice is associated with splenocyte apoptosis and unresponsiveness 6-gingerol prevents patulin-induced genotoxicity in HepG2 cells Nesprin-1 and -2 are involved in the pathogenesis of Emery Dreifuss muscular dystrophy and are critical for nuclear envelope integrity Mendes-Giannini MJS and Fusco-Almeida AM (2016) An Intracellular Arrangement of Histoplasma capsulatum Yeast-Aggregates Generates Nuclear Damage to the Cultured Murine Alveolar Macrophages Received: 09 January 2015; Accepted: 18 December 2015; Published: 11 January 2016 Copyright © 2016 Pitangui, Sardi, Voltan, dos Santos, da Silva, da Silva, Souza, Soares, Rodríguez-Arellanes, Taylor, Mendes-Giannini and Fusco-Almeida. 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Fusco-Almeida, YW5hLm1hcmlzYUB1b2wuY29tLmJy 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 The dates displayed for an article provide information on when various publication milestones were reached at the journal that has published the article activities on preceding journals at which the article was previously under consideration are not shown (for instance submission All content on this site: Copyright © 2025 Elsevier B.V. Jaguar will issue 6,331,713 common shares to AGEM Canada-based Jaguar Mining has agreed to acquire the Pitangui Project and the remaining stake in the Acurui Project in Brazil from IAMGOLD subsidiary AGEM via a share purchase agreement Jaguar will acquire the assets by issuing 6,331,713 common shares to AGEM The shares have an aggregate value of $9m (C$12.04m) Jaguar will also grant a net smelter returns royalty on the projects to AGEM the transaction is due to be closed later this month Located 110km north-west of Belo Horizonte in Minas Gerais the Pitangui Project comprises mineral exploration licences It also includes licence applications covering the Archean-aged Pitangui greenstone belt located near the Jaguar Mining Iron Quadrangle area which comprises “world-class multimillion-ounce gold deposits” Don’t let policy changes catch you off guard Stay proactive with real-time data and expert analysis Currently an exploration joint venture between Jaguar and IAMGOLD the Acurui Project comprises exploration tenements near Jaguar’s Paciência plant in the Iron Quadrangle Jaguar Mining president and CEO Vern Baker said: “We are very pleased to announce that we have reached this agreement to purchase IAMGOLD’s Brazilian Assets which is close to our Turmalina Complex (MTL); and the Acurui Tenements Package contiguous with our Paciencia Complex “This transaction furthers our corporate strategy to leverage our extensive existing infrastructure to drive production growth via increased plant throughput The incorporation of the Pitangui mineral property adds approximately 9,000 kilohectares of prospective mining and exploration tenements to our Iron Quadrangle portfolio.” Give your business an edge with our leading industry insights View all newsletters from across the GlobalData Media network Please enable JS and disable any ad blocker It looks like nothing was found at this location Updated 2 May 2024: I removed the reference to Route53 Alias that was incorrectly referred as a chain you can configure your DNS Firewall to automatically trust all domains in a resolution chain (such as aCNAMEor DNAMEchain) Let’s walk through this in nontechnical terms for those unfamiliar with DNS Why use DNS Firewall? DNS Firewall provides protection for outbound DNS requests from your private network in the cloud (Amazon Virtual Private Cloud (Amazon VPC)). These requests route through Amazon Route 53 Resolver for domain name resolution Firewall administrators can configure rules to filter and regulate the outbound DNS traffic DNS Firewall helps to protect against multiple security risks Let’s imagine a malicious actor managed to install and run some code on your Amazon Elastic Compute Cloud (Amazon EC2) instances or containers running inside one of your virtual private clouds (VPCs) The malicious code is likely to initiate outgoing network connections It might do so to connect to a command server and receive commands to execute on your machine Or it might initiate connections to a third-party service in a coordinated distributed denial of service (DDoS) attack It might also try to exfiltrate data it managed to collect on your network your network and security groups are correctly configured They block all outgoing traffic except the one to well-known API endpoints used by your app So far so good—the malicious code cannot dial back home using regular TCP or UDP connections The malicious code may send DNS requests to an authoritative DNS server they control to either send control commands or encoded data and it can receive data back in the response I’ve illustrated the process in the following diagram you can use a DNS Firewall to monitor and control the domains that your applications can query You can deny access to the domains that you know to be bad and allow all other queries to pass through you can deny access to all domains except those you explicitly trust What is the challenge with CNAME and DNAME records Imagine you configured your DNS Firewall to allow DNS queries only to specific well-known domains and blocked all others Your application communicates with alexa.amazon.com; therefore you created a rule allowing DNS traffic to resolve that hostname the DNS system has multiple types of records A similar redirection mechanism happens when resolving DNAME records To allow the complete resolution of such a CNAME chain you could be tempted to configure your DNS Firewall rule to allow all names under amazon.com (*.amazon.com) but that would fail to resolve the last CNAME that goes to cloudfront.net the DNS CNAME chain is controlled by the service your application connects to forcing you to manually maintain the list of rules and authorized domains inside your DNS Firewall rules This parameter allows firewall administrators to only allow the domain your applications query The firewall will automatically trust all intermediate domains in the chain until it reaches the A record with the IP address Let’s see it in action I start with a DNS Firewall already configured with a domain list, a rule group, and a rule that ALLOW queries for the domain alexa.amazon.com The rule group is attached to a VPC where I have an EC2 instance started When I connect to that EC2 instance and issue a DNS query to resolve alexa.amazon.com it only returns the first name in the domain chain (pitangui.amazon.com) and stops there This is expected because pitangui.amazon.com is not authorized to be resolved I update the firewall rule to trust the entire redirection chain I use the AWS CLI to call the update-firewall-rule API with a new parameter firewall-domain-redirection-action set to TRUST_REDIRECTION_DOMAIN The following diagram illustrates the setup at this stage network administrators now have an easy way to implement a strategy to block all domains and authorize only known domains in their DNS Firewall without having to care about CNAMEor DNAMEchains This capability is available at no additional cost in all AWS Regions. Try it out today Seb has been writing code since he first touched a Commodore 64 in the mid-eighties He inspires builders to unlock the value of the AWS cloud Food Research InternationalCitation Excerpt :This could be due to the non-starch complex carbohydrates in yam (Padhan and Panda Stabilizing the pH after five days of fermentation is of interest from an industrial point of view in addition to helping to inhibit undesirable microorganisms (de Miranda et al. A higher concentration of carbohydrates was detected at the beginning of fermentation