Sediment source fingerprints of natural processes and anthropogenic pressures: A contribution to manage the Paraopeba River basin impacted by the B1 tailings dam collapse.

Understanding the origins of sediment transport in river systems is crucial for effective watershed management, especially after catastrophic events. This information is essential for the development of integrated strategies that guarantee water security in river basins. The present study aimed to investigate the rupture of the B1 tailings dam of the Córrego do Feijão mine, which drastically affected the Brumadinho region (Minas Gerais, Brazil). To address this issue, a confluence-based sediment fingerprinting approach was developed through the SedSAT model. Uncertainty was assessed through Monte Carlo simulations and Mean Absolute Error (MAE). Estimates of the overall average contributions of each tributary were quantified for each station and annually during the period 2019-2021. It was observed that the sampling point PT-09, closest to the dam breach, contributed to almost 80% of the Paraopeba River in 2019. Despite the dredging efforts, this percentage increased to 90% in 2020 due to the need to restore the highly degraded area. Additionally, the main tributaries contributing to sediment increase in the river are Manso River "TT-03" (almost 36%), associated with an area with a high percentage of urban land use, and Cedro stream "TT-07" (almost 71%), whose geology promotes erosion, leading to higher sediment concentration. Uncertainties arise from the limited number of available tracers, variations caused by dredging activities, and reduced data in 2020 due to the pandemic. Parameters such as land use, riparian vegetation degradation, downstream basin geology, and increased precipitation are key factors for successfully assessing tributary contributions to the Paraopeba River. The obtained results are promising for a preliminary analysis, allowing the quantification of key areas due to higher erosion and studying how this disaster affected the watershed. This information is crucial for improving decision-making, environmental governance, and the development of mitigating measures to ensure water security. This study is pioneering in evaluating this methodology in watersheds affected by environmental disasters, where restoration efforts are ongoing.

Beyond land use planning and ecosystem services assessment with the conservation use potential framework: A study in the Upper Rio das Velhas basin, Brazil.

Human actions can damage the ecosystems and affect the services depending on them, with ample detrimental consequences. In earlier studies, the Conservation Use Potential (PCU) framework proved useful in assessing the capacity for aquifer recharge, suitable land uses and resistance to erosion at the river basin scale. On the other hand, the joint analysis of PCU and land uses allowed identifying the adequacy of current uses in relation to suitability (natural uses) in various basins. This was especially useful from the management perspective in basins with environmental conflicts, where current uses differed from suitability, because the PCU indicated how and where the conflicts should be mitigated. Besides the use as management tool, the PCU has potential to shed light over environmental issues such as ecosystem services, but that was not tempted so far. The aim of this work was therefore to bridge that knowledge gap and frame the PCU's application from the standpoint of Ecosystem Services (ES) assessment. We demonstrated how the PCU could be used to improve provision (recharge), support (sustainable agriculture) and regulation (resistance to erosion) services in a specific basin with land use conflicts (the Upper Rio das Velhas basin, located in Minas Gerais, Brazil), through the planning of suitable uses. It was noted that the studied basin is mostly composed of Very Low, Low and Medium potentials. These classes occur because steep slopes, fragile soils and lithologies with high denudation potential and low nutrient supply dominate in the basin. On the other hand, urban sprawl has a negative impact on all ES, while maintaining agricultural areas with appropriate management can effectively regulate erosion. As per the current results, the premise of using the PCU as joint management-environmental tool was fully accomplished, and is recommended a basis for public policy design and implementation in Brazil and elsewhere.

The secretome of macrophages has a differential impact on spinal cord injury recovery according to the polarization protocol.

Introduction: The inflammatory response after spinal cord injury (SCI) is an important contributor to secondary damage. Infiltrating macrophages can acquire a spectrum of activation states, however, the microenvironment at the SCI site favors macrophage polarization into a pro-inflammatory phenotype, which is one of the reasons why macrophage transplantation has failed. Methods: In this study, we investigated the therapeutic potential of the macrophage secretome for SCI recovery. We investigated the effect of the secretome in vitro using peripheral and CNS-derived neurons and human neural stem cells. Moreover, we perform a pre-clinical trial using a SCI compression mice model and analyzed the recovery of motor, sensory and autonomic functions. Instead of transplanting the cells, we injected the paracrine factors and extracellular vesicles that they secrete, avoiding the loss of the phenotype of the transplanted cells due to local environmental cues. Results: We demonstrated that different macrophage phenotypes have a distinct effect on neuronal growth and survival, namely, the alternative activation with IL-10 and TGF-ß1 (M(IL-10+TGF-ß1)) promotes significant axonal regeneration. We also observed that systemic injection of soluble factors and extracellular vesicles derived from M(IL-10+TGF-ß1) macrophages promotes significant functional recovery after compressive SCI and leads to higher survival of spinal cord neurons. Additionally, the M(IL-10+TGF-ß1) secretome supported the recovery of bladder function and decreased microglial activation, astrogliosis and fibrotic scar in the spinal cord. Proteomic analysis of the M(IL-10+TGF-ß1)-derived secretome identified clusters of proteins involved in axon extension, dendritic spine maintenance, cell polarity establishment, and regulation of astrocytic activation. Discussion: Overall, our results demonstrated that macrophages-derived soluble factors and extracellular vesicles might be a promising therapy for SCI with possible clinical applications.

A framework model to integrate sources and pathways in the assessment of river water pollution.

Metal and nutrient pollution, soil erosion, and alterations in climate and hydrology are prevalent issues that impact the water quality of riverine systems. However, integrated approaches to assess and isolate causes and paths of river water pollution are scarce, especially in the case of watersheds impacted by multiple hazardous activities. Therefore, a framework model for investigating the multiple sources of river water pollution was developed. The chosen study area was the Paraopeba River basin located in the Minas Gerais, Brazil. Besides multiple agriculture, industrial, and urban pollution sources, this region was profoundly affected by the rupture of the B1 tailings dam (in January 2019) at the Córrego do Feijão mine, resulting in the release of metal-rich waste. Considering this situation, thirty-nine physicochemical and hydromorphological parameters were examined in the Paraopeba River basin, in the 2019-2023 period. The analysis involved various statistical techniques, including bivariate and multivariate methods such as correlation analysis, principal component analysis, and clustering. The Paraopeba River was mainly impacted by metal contamination resulting from the dam collapse, whereas nutrient contamination, mainly from urban and industrial discharges, predominantly affected its tributaries. Additionally, the elevated concentrations of aluminum, iron, nitrate, and sulfate in both main river and tributaries can be attributed to diffuse and point source pollution. In terms of hydromorphology and soil type, the interaction between woody vegetation and erosion-resistant soils, especially latosols, contributes to the stability of riverbanks in the main river. Meanwhile, in the tributaries, the presence of neosols and sparse vegetation in urbanized areas promoted riverbank erosion potentially amplifying pollution. While the study was conducted in a particular watershed, the findings are based on a methodology that can be applied universally. Hence, the insights on surface water quality from this research can be a valuable resource for researchers studying watersheds with diverse pollution sources.

Assessing the uptake of incentivised physical health checks for people with serious mental illness.

BACKGROUND: People with serious mental illness (SMI) are more likely to suffer from physical illnesses. The onset of many of these illnesses can be prevented if detected early. Physical health screening for people with SMI is incentivised in primary care in England through the Quality and Outcomes Framework (QOF). General Practitioners are paid to conduct annual physical health checks (PHCs) on their SMI patients, including checks on body mass index (BMI), cholesterol, and alcohol consumption. AIM: To assess the impact of removing and reintroducing QOF financial incentives on uptake of three PHCs (BMI, cholesterol, and alcohol consumption) for patients with SMI. DESIGN AND SETTING: Cohort study using UK primary care data from the Clinical Practice Research Datalink between April 2011 and March 2020. METHOD: We employed a difference-in-difference analysis to compare differences in the uptake before and after the intervention accounting for relevant observed and unobserved confounders. RESULTS: We found an immediate change in uptake after PHCs were removed from, and after they were added back to the QOF list. For BMI, cholesterol, and alcohol checks the overall impact of removal was a reduction in uptake of 14.3, 6.8, and 11.9 percentage points, respectively. The reintroduction of BMI screening in the QOF increased the uptake by 10.2 percentage points. CONCLUSION: Our analysis supports the hypothesis that QOF incentives lead to better uptake of PHCs.

Proteomic profiles of Leptospira borgpetersenii serovar Hardjo strains JB197 and HB203 cultured at different temperatures.

Leptospirosis is a global zoonotic disease affecting humans, domestic, and wild animals. Leptospira are typically shed in the urine of reservoir hosts which persist in suitable environments where incidental host transmission occurs after direct contact with infected urine or contaminated environments. Interestingly, serologically identical L. borgpetersenii serovar Hardjo strains JB197 and HB203 show divergent disease severity in the hamster model; JB197 causes severe acute infection while HB203 causes persistent chronic infection. Historically, serovar Hardjo was limited to culture at 29 °C, but utilization of HAN media allows propagation from host tissues at 37 °C. Here, the proteome of strains JB197 and HB203 were characterized after culture from experimentally challenged hamsters at 29 °C and 37 °C. Comparative analyses of JB197 and HB203 samples cultured at 29 °C yielded 425 significantly differentially expressed (DE) proteins, while strains at 37 °C yielded 613 DE proteins including prominent outer membrane proteins and known virulence factors. In agreement, membrane protein GO terms were identified by STRING network analyses along with numerous metabolic KEGG pathways consistent with condition differences. Within strain, JB197 cultured at 29 °C vs 37 °C identified 529 DE proteins, while HB203 identified 524 DE proteins. Investigating differential protein profiles provide insights into strain specific behaviors with implications for better understanding host-pathogen interactions, disease transmission, and response to environmental conditions which can contribute to vaccine development, diagnostic improvement, and ultimately leptospirosis control. SIGNIFICANCE: Leptospirosis is a devastating zoonotic disease affecting humans, wild and domestic animals around the globe. Different species and serovars of Leptospira can affect various animal host species differently; for instance, a serovar that is asymptomatic in the rat may cause severe disease in a dog or human. These differences in host response are not only found at the species and serovar level for Leptospira, but also at the strain level. A prime example comes from strains JB197 and HB203, both species L. borgpetersenii, both serovar Hardjo. Interestingly, JB197 causes a severe acute infection in the hamster while HB203 causes an asymptomatic chronic infection. Understanding these unique relationships between pathogen and host species is important, especially in the context of prevention technologies such as vaccine design, where the strain of Leptospira used as a bacterin might have different efficiencies in different hosts. In this study, proteomic profiles of strains JB197 and HB203 were analyzed, and results revealed diverse protein expression profiles of outer membrane proteins, as well as proteins functioning in motility and growth.

Chronic Lithium Intoxication: A Challenging Diagnosis.

Lithium has been used in clinical practice since the 1970s. This medication is commonly used to treat and prevent bipolar disorder, but it has a narrow therapeutic index, making toxicity a frequent occurrence. Chronic lithium intoxication can arise due to progressive accumulation, particularly in contexts of dehydration. The effects of chronic lithium intoxication on the nervous, renal, and cardiac systems, as well as on the thyroid and parathyroid glands, are well documented in the literature. The authors present the case of a 66-year-old woman with schizoaffective psychosis and chronic kidney disease, admitted due to altered mental status and dysarthria. Notwithstanding an earlier clinical recommendation to cease lithium administration more than a year ago, the patient continued its usage, culminating in neurological, cardiac, renal, and endocrine manifestations. Although the diagnosis was delayed, her clinical progression was favorable, obviating the need for renal replacement therapy. This case highlights the importance of a detailed medical history and the diagnostic challenges in clinical practice. The use of this drug without proper monitoring can lead to multisystem dysfunction.

Optimized reconstruction of the absorption spectra of kidney tissues from the spectra of tissue components using the least squares method.

With the objective of developing new methods to acquire diagnostic information, the reconstruction of the broadband absorption coefficient spectra (µa [λ]) of healthy and chromophobe renal cell carcinoma kidney tissues was performed. By performing a weighted sum of the absorption spectra of proteins, DNA, oxygenated, and deoxygenated hemoglobin, lipids, water, melanin, and lipofuscin, it was possible to obtain a good match of the experimental µa (λ) of both kidney conditions. The weights used in those reconstructions were estimated using the least squares method, and assuming a total water content of 77% in both kidney tissues, it was possible to calculate the concentrations of the other tissue components. It has been shown that with the development of cancer, the concentrations of proteins, DNA, oxygenated hemoglobin, lipids, and lipofuscin increase, and the concentration of melanin decreases. Future studies based on minimally invasive spectral measurements will allow cancer diagnosis using the proposed approach.

Dipyrone (Metamizole)-Induced Stevens-Johnson Syndrome.

Stevens-Johnson Syndrome (SJS), a severe mucocutaneous hypersensitivity reaction primarily triggered by drugs, poses a low-incidence, high-mortality challenge. This report explores its clinical nuances and emphasizes supportive care as the mainstay of treatment. A 74-year-old female, burdened with a complex medical history, presented with a non-pruritic macular rash escalating to skin and oral mucosal involvement. A recent introduction of dipyrone (metamizole) implicated drug-induced SJS. Histopathological confirmation guided treatment involving supportive care, corticosteroids, and wound care, resulting in clinical improvement. The case underscores the significance of histopathological confirmation and thorough medication history in navigating SJS complexities, especially in patients with comorbidities like connective tissue disease. A successful multidisciplinary approach and the decision for post-discharge monitoring highlight the intricate management challenges. This case illuminates the intricate interplay of medication-induced hypersensitivity, comorbidities, and management challenges in SJS. Optimal outcomes require prompt diagnosis, trigger identification, and a multidisciplinary treatment approach, emphasizing ongoing research and clinical vigilance.

From Suspected COVID-19 to Anti-synthetase Syndrome: A Diagnostic Challenge in the Pandemic Era.

Anti-synthetase syndrome (ASS), a rare immunomediated disease, is characterized by multiple signs and symptoms. Not all patients develop the entire clinical spectrum of the syndrome, as it often varies depending on the involved antibodies. In this case report, a 53-year-old non-smoking woman had complaints of fatigue and dyspnea on exertion for five weeks. The outpatient study revealed creatine kinase (CK) 351U/L, ANAs+, anti-SSa+, normal echocardiogram, and a chest X-ray suggesting imaging suspicion of SARS-CoV-2 pneumonia. Referred to the emergency department, she was hospitalized for bilateral interstitial pneumonia without respiratory failure. Three SARS-CoV-2 polymerase chain reaction tests were negative. She underwent a five-day course of dexamethasone 6mg due to suspected coronavirus disease 2019 (COVID-19) sequelae with favorable progress. About a month later, she experienced fatigue, exertional intolerance, morning cough, and Raynaud's phenomenon episodes. Anti-SARS-CoV-2 antibodies were negative, and a follow-up chest CT showed bilateral organizing pneumonia. Bronchofibroscopy and bronchoalveolar lavage with cytology suggestive of inflammatory appearance, predominantly CD8+ lymphocytes, were performed. Subsequently, positive results for anti-OJ antibodies were obtained. A diagnosis of ASS was established, and prednisolone was initiated at 60mg/day with a tapering regimen, resulting in clinical and radiological improvement. Additional therapy with azathioprine was proposed. This case is presented due to highly suggestive COVID-19 imaging changes, emphasizing the importance of a high suspicion of ASS, despite nearly exclusive pulmonary involvement, with only one isolated elevated CK value and no musculoskeletal complaints. It is also noteworthy for the association with anti-OJ antibodies, rarely identified, often presenting interstitial lung disease as an isolated manifestation.

Explaining Bounding Boxes in Deep Object Detectors Using Post Hoc Methods for Autonomous Driving Systems.

Deep learning has rapidly increased in popularity, leading to the development of perception solutions for autonomous driving. The latter field leverages techniques developed for computer vision in other domains for accomplishing perception tasks such as object detection. However, the black-box nature of deep neural models and the complexity of the autonomous driving context motivates the study of explainability in these models that perform perception tasks. Moreover, this work explores explainable AI techniques for the object detection task in the context of autonomous driving. An extensive and detailed comparison is carried out between gradient-based and perturbation-based methods (e.g., D-RISE). Moreover, several experimental setups are used with different backbone architectures and different datasets to observe the influence of these aspects in the explanations. All the techniques explored consist of saliency methods, making their interpretation and evaluation primarily visual. Nevertheless, numerical assessment methods are also used. Overall, D-RISE and guided backpropagation obtain more localized explanations. However, D-RISE highlights more meaningful regions, providing more human-understandable explanations. To the best of our knowledge, this is the first approach to obtaining explanations focusing on the regression of the bounding box coordinates.

First Genome Sequence of the Microcolonial Black Fungus Saxispiralis lemnorum MUM 23.14: Insights into the Unique Genomic Traits of the Aeminiaceae Family.

Saxispiralis lemnorum MUM 23.14 is an extremotolerant microcolonial black fungus, originally isolated from a biodeteriorated limestone artwork in Portugal. This recently introduced species belongs to the Aeminiaceae family, representing the second member of this monophyletic clade. This fungus exhibits a unique set of characteristics, including xerophily, cold tolerance, high UV radiation tolerance, and an exceptional ability to thrive in NaCl concentrations of up to 30% while also enduring pH levels ranging from 5 to 11. To gain insights into its genomic traits associated with stress resistance mechanisms, specialization, and their potential implications in stone biodeterioration, we conducted a comprehensive genome sequencing and analysis. This draft genome not only marks the first for the Saxispiralis genus but also the second for the Aeminiaceae family. Furthermore, we performed two comparative genomic analyses: one focusing on the closest relative within the Aeminiaceae family, Aeminium ludgeri, and another encompassing the genome of different extremotolerant black fungi. In this study, we successfully achieved high genome completeness for S. lemnorum and confirmed its close phylogenetic relationship to A. ludgeri. Our findings revealed traits contributing to its extremophilic nature and provided insights into potential mechanisms contributing to stone biodeterioration. Many traits are common to both Aeminiaceae species and are shared with other black fungi, while numerous unique traits may be attributed to species-specific characteristics.

Geomorphologic risk zoning to anticipate tailings dams' hazards: A study in the Brumadinho's mining area, Minas Gerais, Brazil.

The use of tailings dams in the mining industry is recurrent and a matter of concern given the risk of collapse. The planning of tailings dam's emplacement usually attends construction design criteria and site geotechnical properties, but often neglects the risk of installing the depositional facilities in potentially unstable landscapes, namely those characterized by steep slopes and(or) high drainage densities. In order to help bridging this gap, the present study developed a framework model whereby geomorphologic vulnerability is assessed by a set of morphometric parameters (e.g., drainage density; relief ratio; roughness coefficient). Using the Ribeirão Ferro-Carvão micro-basin (3265.16 ha) as test site, where six dams currently receive tailings from the mining of iron-ore deposits in the Brumadinho region (Minas Gerais, Brazil) and one has collapsed in 25 January 2019 (the B1 dam of Córrego do Feijão mine of Vale, S.A.), the risk of dam instability derived from geomorphologic vulnerability was assessed and alternative suitable locations were highlighted when applicable. The results made evident the location of five dams (including the collapsed B1) in high-risk regions and two in low-risk regions, which is preoccupying. The alternative locations represent 58 % of Ribeirão Ferro-Carvão micro-basin, which is a reasonable and workable share. Overall, the study exposed the fragility related with tailings dams' geography, which is not restricted to the studied micro-basin, because dozens of active tailings dams exist in the parent basin (the Paraopeba River basin) that can also be vulnerable to geomorphologically-dependent hydrologic hazards such as intensive erosion, valley incision or flash floods. Attention to this issue is therefore urgent to prevent future tragedies related with tailings dams' breaks, in the Paraopeba River basin or elsewhere, using the proposed framework model as guide.

LipL41 and LigA/LigB Gene Silencing on a LipL32 Knockout Leptospira interrogans Reveals the Impact of Multiple Mutations on Virulence.

Leptospirosis is a global zoonosis caused by pathogenic bacteria of the genus Leptospira. The application of the CRISPR/Cas9 system has facilitated the generation of mutants and subsequent evaluation of phenotypes. Since DNA breaks induced by RNA-guided Cas9 nuclease are lethal to Leptospira, different methodologies were implemented to overcome this limitation. Initially, CRISPR interference (CRISPRi) was employed to create knockdown mutants, utilizing a catalytically inactive Cas9 (dCas9). Subsequently, the co-expression of CRISPR/Cas9 and a DNA repair system from Mycobacterium smegmatis enabled the generation of scarless knockout mutants. We eliminated plasmids from the lipL32 knockout L. interrogans strain and further achieved multiple gene mutations via gene silencing in this knockout background. Strains lacking both LipL41 and LipL32 and LigA, LigB, and LipL32, were evaluated. The absence of proteins LipL32 and LipL41 had no effect on leptospiral virulence. On the other hand, mutants lacking LigA, LigB, and LipL32 were unable to cause acute disease. The expanded apparatus for genetic manipulation of pathogenic leptospires via the CRISPR/Cas9 system has allowed the evaluation of multiple mutations upon leptospiral virulence. This work shows that LipL32 and LipL41 are not required for acute disease and consolidates LigA and LigB proteins as virulence factors.

Concurrent colonization of rodent kidneys with multiple species and serogroups of pathogenic Leptospira.

Rodents are important reservoir hosts of pathogenic leptospires in the US Virgin Islands. Our previous work determined that trapped rodents were colonized with Leptospira borgpetersenii serogroup Ballum (n = 48) and/or Leptospira kirschneri serogroup Icterohaemorrhagiae (n = 3). In addition, nine rodents appeared to be colonized with a mixed population comprising more than one species/serogroup. The aim of this study was to validate this finding by characterizing clonal isolates derived from cultures of mixed species. Cultures of presumptive mixed species (designated LR1, LR5, LR37, LR57, LR60, LR61, LR68, LR70, and LR72) were propagated in different media including Hornsby-Alt-Nally (HAN) media, incubated at both 29℃ and 37℃, and T80/40/LH incubated at 29℃. Polyclonal reference antisera specific for serogroup Ballum and Icterohaemorrhagiae were used to enrich for different serogroups followed by subculture on agar plates. Individual colonies were then selected for genotyping and serotyping. Of the nine cultures of mixed species/serogroups, a single clonal isolate was separated in five of them: L. borgpetersenii serogroup Ballum in LR1, LR5, and LR37, and L. kirschneri serogroup Icterohaemorrhagiae in LR60 and LR72. In four of the cultures with mixed species (LR57, LR61, LR68, and LR70), clonal isolates of both L. borgpetersenii serogroup Ballum and L. kirschneri serogroup Icterohaemorrhagiae were recovered. Our results definitively establish that rodents can be colonized with more than one species/serogroup of Leptospira concurrently. The identification and characterization of multiple species/serogroups of Leptospira from individual reservoir hosts of infection are essential to understand the epidemiology and transmission of disease to both human and domestic animal populations.IMPORTANCEPathogenic Leptospira, the causative agent of human and animal leptospirosis, comprise a diverse genus of species/serogroups which are inherently difficult to isolate from mammalian hosts due to fastidious growth requirements. Molecular evidence has indicated that reservoir hosts of Leptospira may shed multiple species concurrently. However, evidence of this phenomena by culture has been lacking. Culture is definitive and is essential for comprehensive characterization of recovered isolates by high-resolution genome sequencing and serotyping. In this work, a protocol using recently developed novel media formulations, in conjunction with reference antisera, was developed and validated to demonstrate the recovery of multiple species/serogroups of pathogenic Leptospira from the same host. The identification and characterization of multiple species/serogroups of Leptospira from individual reservoir hosts of infection are essential to understand the epidemiology and transmission of disease to both human and domestic animal populations.

Recombinant antigen 5 from Polybia paulista wasp venom (Hymenoptera, Vespidae): Antigen-specific antibody production and functional profile of CD4+ T cells in the immune response.

Polybia paulista is a neotropical social wasp related to severe accidents and allergic reactions cases, including anaphylaxis, in southeastern Brazil. Antigen 5 (Poly p 5) is a major allergenic protein from its venom with potential use for component-resolved diagnostic. Therefore, the previous characterization of the immune response profile triggered by Poly p 5 should be evaluated. Recombinant Poly p 5 (rPoly p 5) was used to sensitize BALB/c mice with six weekly intradermal doses, and the specific antibody production and the functional profile of CD4+ T cells were assessed. rPoly p 5 induced the production of specific immunoglobulins (sIg) sIgE, sIgG1 and sIgG2a, which could recognize natural Poly p 5 presented in the venom of four different wasp species. rPoly p 5 stimulated in vitro the CD4+ T cells from immunized mice, which showed a significant proliferative response. These antigen-specific CD4+T cells produced IFN-γ and IL-17A cytokines and increased ROR-γT transcription factor expression. No differences between the control group and sensitized mice were found in IL-4 production and GATA-3 and T-bet expression. Interestingly, increased CD25+FoxP3+ regulatory T cells (Tregs) frequency was observed in the splenocyte cell cultures from rPoly p 5 immunized mice after the in vitro stimulation with both P. paulista venom extract and rPoly p 5. Here we showed that rPoly p 5 induces antigen-specific antibodies capable of recognizing Antigen 5 in the venom of four wasp species and modulates antigen-specific CD4+ T cells to IFN-γ production response associated with a Th17 profile in sensitized mice. These findings emphasize the potential use of rPoly p 5 as an essential source of a major wasp allergen with significant immunological properties.

Expanding the Microcolonial Black Fungi Aeminiaceae Family: Saxispiralis lemnorum gen. et sp. nov. (Mycosphaerellales), Isolated from Deteriorated Limestone in the Lemos Pantheon, Portugal.

With an impressive ability to survive in harsh environments, black fungi are an ecological group of melanized fungi that are widely recognized as a major contributor to the biodeterioration of stone cultural heritage materials. As part of the ongoing efforts to study the fungal diversity thriving in a deteriorated limestone funerary art piece at the Lemos Pantheon, a national monument located in Águeda, Portugal, two isolates of an unknown microcolonial black fungus were retrieved. These isolates were thoroughly studied through a comprehensive analysis based on a multi-locus phylogeny of a combined dataset of ITS rDNA, LSU, and rpb2, along with morphological, physiological, and ecological characteristics. Based on the data obtained from this integrative analysis, we propose a new genus, Saxispiralis gen. nov., and a new species, Saxispiralis lemnorum sp. nov., in the recently described Aeminiaceae family (order Mycosphaerellales). Prior to this discovery, this family only had one known genus and species, Aeminium ludgeri, also isolated from deteriorated limestone. Additionally, considering the isolation source of the fungus and to better understand its potential contribution to the overall stone monument biodeterioration, its in vitro biodeteriorative potential was also evaluated. This work represents a significant contribution to the understanding of the fungal diversity involved in the biodeterioration of limestone heritage.

Evaluation of Leptospira interrogans knockdown mutants for LipL32, LipL41, LipL21, and OmpL1 proteins.

Introduction: Leptospirosis is a worldwide zoonosis caused by pathogenic and virulent species of the genus Leptospira, whose pathophysiology and virulence factors remain widely unexplored. Recently, the application of CRISPR interference (CRISPRi) has allowed the specific and rapid gene silencing of major leptospiral proteins, favoring the elucidation of their role in bacterial basic biology, host-pathogen interaction and virulence. Episomally expressed dead Cas9 from the Streptococcus pyogenes CRISPR/Cas system (dCas9) and single-guide RNA recognize and block transcription of the target gene by base pairing, dictated by the sequence contained in the 5' 20-nt sequence of the sgRNA. Methods: In this work, we tailored plasmids for silencing the major proteins of L. interrogans serovar Copenhageni strain Fiocruz L1-130, namely LipL32, LipL41, LipL21 and OmpL1. Double- and triple-gene silencing by in tandem sgRNA cassettes were also achieved, despite plasmid instability. Results: OmpL1 silencing resulted in a lethal phenotype, in both L. interrogans and saprophyte L. biflexa, suggesting its essential role in leptospiral biology. Mutants were confirmed and evaluated regarding interaction with host molecules, including extracellular matrix (ECM) and plasma components, and despite the dominant abundance of the studied proteins in the leptospiral membrane, protein silencing mostly resulted in unaltered interactions, either because they intrinsically display low affinity to the molecules assayed or by a compensation mechanism, where other proteins could be upregulated to fill the niche left by protein silencing, a feature previously described for the LipL32 mutant. Evaluation of the mutants in the hamster model confirms the augmented virulence of the LipL32 mutant, as hinted previously. The essential role of LipL21 in acute disease was demonstrated, since the LipL21 knockdown mutants were avirulent in the animal model, and even though mutants could still colonize the kidneys, they were found in markedly lower numbers in the animals' liver. Taking advantage of higher bacterial burden in LipL32 mutant-infected organs, protein silencing was demonstrated in vivo directly in leptospires present in organ homogenates. Discussion: CRISPRi is now a well-established, attractive genetic tool that can be applied for exploring leptospiral virulence factors, leading to the rational for designing more effective subunit or even chimeric recombinant vaccines.

Payment for Environmental Services: A critical review of schemes, concepts, and practice in Brazil.

The Payment for Environmental Services (PES) is often conceived through complex schemes without a clear definition of all concepts involved. This study presents the results of a systematic literature review on PES schemes in Brazil, accompanied by a critical assessment of their efficacy for potential environmental gains. The PES approaches were grouped into six categories based on the research focus, and those that were focused on PES policies were identified as the most studied. A particular emphasis has been given to the Amazon and Atlantic Forest biomes, where the ecosystem services studied were mostly centered on issues related to carbon and water, respectively. Approximately one-third of all schemes provided no clear definition of which ecosystem services are proposed for payment. In addition, the review showed no consensus among studies on the definition of services in similar schemes. Most schemes presented no payment system conditioned on the provision of environmental services. Furthermore, the review showed that the absence of clarity in the application of concepts may hinder the development of public policies to properly implement PES in Brazil. The conclusion is that standardizing terms used in the literature and in PES schemes is critical; therefore, the use of the Common International Classification of Ecosystem Services (CICES) as a reference is recommended to ensure clarity, objectivity and, more importantly, the expected environmental efficacy.