S. aureus drives itch and scratch-induced skin damage through a V8 protease-PAR1 axis.

Itch is an unpleasant sensation that evokes a desire to scratch. The skin barrier is constantly exposed to microbes and their products. However, the role of microbes in itch generation is unknown. Here, we show that Staphylococcus aureus, a bacterial pathogen associated with itchy skin diseases, directly activates pruriceptor sensory neurons to drive itch. Epicutaneous S. aureus exposure causes robust itch and scratch-induced damage. By testing multiple isogenic bacterial mutants for virulence factors, we identify the S. aureus serine protease V8 as a critical mediator in evoking spontaneous itch and alloknesis. V8 cleaves proteinase-activated receptor 1 (PAR1) on mouse and human sensory neurons. Targeting PAR1 through genetic deficiency, small interfering RNA (siRNA) knockdown, or pharmacological blockade decreases itch and skin damage caused by V8 and S. aureus exposure. Thus, we identify a mechanism of action for a pruritogenic bacterial factor and demonstrate the potential of inhibiting V8-PAR1 signaling to treat itch.

Higher-Dose Primaquine to Prevent Relapse of Plasmodium vivax Malaria.

BACKGROUND: In most of the Americas, the recommended treatment to prevent relapse of Plasmodium vivax malaria is primaquine at a total dose of 3.5 mg per kilogram of body weight, despite evidence of only moderate efficacy. METHODS: In this trial conducted in Brazil, we evaluated three primaquine regimens to prevent relapse of P. vivax malaria in children at least 5 years of age and in adults with microscopy-confirmed P. vivax monoinfection. All the patients received directly observed chloroquine for 3 days (total dose, 25 mg per kilogram). Group 1 received a total primaquine dose of 3.5 mg per kilogram (0.5 mg per kilogram per day) over 7 days with unobserved administration; group 2 received the same regimen as group 1 but with observed administration; and group 3 received a total primaquine dose of 7.0 mg per kilogram over 14 days (also 0.5 mg per kilogram per day) with observed administration. We monitored the patients for 168 days. RESULTS: We enrolled 63 patients in group 1, 96 in group 2, and 95 in group 3. The median age of the patients was 22.4 years (range, 5.4 to 79.8). By day 28, three P. vivax recurrences were observed: 2 in group 1 and 1 in group 2. By day 168, a total of 70 recurrences had occurred: 24 in group 1, 34 in group 2, and 12 in group 3. No serious adverse events were noted. On day 168, the percentage of patients without recurrence was 58% (95% confidence interval [CI], 44 to 70) in group 1, 59% (95% CI, 47 to 69) in group 2, and 86% (95% CI, 76 to 92) in group 3. Survival analysis showed a difference in the day 168 recurrence-free percentage of 27 percentage points (97.5% CI, 10 to 44; P<0.001) between group 1 and group 3 and a difference of 27 percentage points (97.5% CI, 12 to 42; P<0.001) between group 2 and group 3. CONCLUSIONS: The administration of primaquine at a total dose of 7.0 mg per kilogram had higher efficacy in preventing relapse of P. vivax malaria than a total dose of 3.5 mg per kilogram through day 168. (Supported by the U.S. Agency for International Development; ClinicalTrials.gov number, NCT03610399.).

Overcoming pH defenses on the skin to establish infections.

Skin health is influenced by the composition and integrity of the skin barrier. The healthy skin surface is an acidic, hypertonic, proteinaceous, and lipid-rich environment that microorganisms must adapt to for survival, and disruption of this environment can result in dysbiosis and increase risk for infectious diseases. This work provides a brief overview of skin barrier function and skin surface composition from the perspective of how the most common skin pathogen, Staphylococcus aureus, combats acid stress. Advancements in replicating this environment in the laboratory setting for the study of S. aureus pathogenesis on the skin, as well as future directions in this field, are also discussed.

Lianas in tropical dry seasonal forests have a high hydraulic efficiency but not always a higher embolism resistance than lianas in rainforests.

BACKGROUND AND AIMS: Lianas have higher relative abundance and biomass in drier seasonal forests than rainforests, but whether this difference is associated with their hydraulic strategies is unclear. Here, we investigate whether lianas of seasonally dry forests are safer and more efficient in water transport than rainforest ones, explaining liana abundance patterns. METHODS: We measured hydraulic traits on five pairs of congeneric lianas of the tribe Bignonieae in two contrasting forest sites: the wet 'Dense Ombrophilous Forest' in the Central Amazonia (~ 2 dry months) and the drier 'Semideciduous Seasonal Forest' in the inland Atlantic Forest (~6 dry months). We also gathered a broader database, including 197 trees and 58 liana species from different tropical forests, to compare hydraulic safety between habits and forest types. KEY RESULTS: Bignonieae lianas from both forests had high and similar hydraulic efficiency and exhibited variability in resistance to embolism across forest types when phylogenetic relationships are taken into account. Three genera had higher hydraulic safety in the seasonal forest than in the rainforest, but species across both forests had similar positive hydraulic safety margin despite lower predawn water potential values of seasonal forest' lianas. We did not find the safety-efficiency trade-off. Merging our results with previously published data evidenced a high variability of resistance to embolism in both trees and lianas, independent of forest types. CONCLUSIONS: The high hydraulic efficiency of lianas detected here probably favors their rapid growth across tropical forests, but differences in hydraulic safety highlight that some species are highly vulnerable and may rely on other mechanisms to cope with drought. Future research on the lethal dehydration threshold and the connection between hydraulic resistance strategies and liana abundance could offer further insights into tropical forest dynamics under climatic threats.

Imaging features for the evaluation of skin and nail infections.

Clinical manifestations of dermatological and musculoskeletal conditions can sometimes overlap, leading to confusion in diagnosis. Patients with nail and skin infections may undergo imaging examinations with suspicions of muscle, tendon, or joint injuries. Dermatological infections often involve soft tissues and musculoskeletal structures, and their etiology can range from fungi, bacteria, viruses, to protozoa. Relying solely on physical examination may not be sufficient for accurate diagnosis and treatment planning, necessitating the use of complementary imaging exams. The objective of this paper is to present and discuss imaging findings of the main infectious conditions affecting the nail apparatus and skin. The paper also highlights the importance of imaging in clarifying diagnostic uncertainties and guiding appropriate treatment for dermatological conditions.

Localization and interaction studies of the Salmonella enterica ethanolamine ammonia-lyase (EutBC), its reactivase (EutA), and the EutT corrinoid adenosyltransferase.

Some prokaryotes compartmentalize select metabolic capabilities. Salmonella enterica subspecies enterica serovar Typhimurium LT2 (hereafter S. Typhimurium) catabolizes ethanolamine (EA) within a proteinaceous compartment that we refer to as the ethanolamine utilization (Eut) metabolosome. EA catabolism is initiated by the adenosylcobalamin (AdoCbl)-dependent ethanolamine ammonia-lyase (EAL), which deaminates EA via an adenosyl radical mechanism to yield acetaldehyde plus ammonia. This adenosyl radical can be quenched, requiring the replacement of AdoCbl by the ATP-dependent EutA reactivase. During growth on ethanolamine, S. Typhimurium synthesizes AdoCbl from cobalamin (Cbl) using the ATP:Co(I)rrinoid adenosyltransferase (ACAT) EutT. It is known that EAL localizes to the metabolosome, however, prior to this work, it was unclear where EutA and EutT localized, and whether they interacted with EAL. Here, we provide evidence that EAL, EutA, and EutT localize to the Eut metabolosome, and that EutA interacts directly with EAL. We did not observe interactions between EutT and EAL nor between EutT and the EutA/EAL complex. However, growth phenotypes of a ΔeutT mutant strain show that EutT is critical for efficient ethanolamine catabolism. This work provides a preliminary understanding of the dynamics of AdoCbl synthesis and its uses within the Eut metabolosome.

The effects of delta-9-tetrahydrocannabinol administration in the regulation of female rat sociosexual behaviour.

By targeting the endocannabinoid system, delta-9-tetrahydrocannabinol (THC) modulates female motivated behaviours, influenced by sex hormones. Both medial preoptic nucleus (MPN) and ventromedial nucleus of the hypothalamus (VMN) are involved in the modulation of female sexual responses. The first triggers proceptivity, whereas the ventrolateral division of the latter (VMNvl) triggers receptivity. These nuclei are modulated by glutamate, which inhibits female receptivity, and GABA, which has a dichotomous action in female sexual motivation. Here, we evaluated the action of THC on the modulation of social and sexual behaviours, on signalling pathways of MPN and VMNvl and how sex hormones influence these parameters. Young ovariectomized female rats, given sex hormones (oestradiol benzoate, EB, and progesterone, P) and THC were used for behavioural testing and for immunofluorescence analyses of vesicular glutamate transporter 2 (VGlut2) and GAD (glutamic acid decarboxylase)67 expression. Results showed that females given EB + P exhibited a higher preference for male partner, as well as higher proceptivity and a higher receptivity than control or females given only EB. Females treated with THC presented similar responses in control or EB + P female rats and even more facilitated behavioural responses in EB females than the ones that did not receive THC. Immunofluorescence results in the MPN exhibited a decreased expression of GAD67 and VGlut2 in EB + THC-treated female rats. Within VMNvl of EB-primed rats no changes in the expression of both proteins were observed after THC exposure. This study demonstrates how the possible outcomes of endocannabinoid system instability within hypothalamic neuron connectivity can modify female rat sociosexual behaviour.

The other side of tropical forest drought: do shallow water table regions of Amazonia act as large-scale hydrological refugia from drought?

Tropical forest function is of global significance to climate change responses, and critically determined by water availability patterns. Groundwater is tightly related to soil water through the water table depth (WT), but historically neglected in ecological studies. Shallow WT forests (WT < 5 m) are underrepresented in forest research networks and absent in eddy flux measurements, although they represent c. 50% of the Amazon and are expected to respond differently to global-change-related droughts. We review WT patterns and consequences for plants, emerging results, and advance a conceptual model integrating environment and trait distributions to predict climate change effects. Shallow WT forests have a distinct species composition, with more resource-acquisitive and hydrologically vulnerable trees, shorter canopies and lower biomass than deep WT forests. During 'normal' climatic years, shallow WT forests have higher mortality and lower productivity than deep WT forests, but during moderate droughts mortality is buffered and productivity increases. However, during severe drought, shallow WT forests may be more sensitive due to shallow roots and drought-intolerant traits. Our evidence supports the hypothesis of neglected shallow WT forests being resilient to moderate drought, challenging the prevailing view of widespread negative effects of climate change on Amazonian forests that ignores WT gradients, but predicts they could collapse under very strong droughts.

O funcionamento da floresta tropical é de importância global para as respostas às mudanças climáticas e é criticamente determinado pelos padrões de disponibilidade de água. A água subterrânea está intimamente relacionada à água do solo através da profundidade do lençol freático, que tem sido historicamente negligenciado em estudos ecológicos. Florestas com lençol freático raso (< 5 m) estão sub-representadas nas redes de pesquisa florestal e ausentes nas medições de fluxo de gases, embora representem ~ 50% da Amazônia e devam responder de forma diferente às secas relacionadas às mudanças globais. Aqui revisamos os padrões de profundidade do lençol freático e suas consequências para plantas, resultados emergentes, e avançamos em um modelo conceitual que integra o ambiente e as distribuições de características funcionais para prever os efeitos das mudanças climáticas. As florestas com lençol freático raso têm uma composição de espécies distinta, com árvores mais aquisitivas na obtenção de recursos e hidrologicamente vulneráveis, dosséis mais baixos e menor biomassa do que as florestas com lençol freático profundo. Durante os anos climáticos 'normais', as florestas com lençol freático raso têm maior mortalidade e menor produtividade do que as florestas com lençol freático profundo, mas durante secas moderadas, a mortalidade é amortecida e a produtividade aumenta. No entanto, durante secas severas, as florestas com lençol freático raso podem ser mais sensíveis devido às raízes superficiais e características funcionais de intolerância à seca. Nossas evidências apoiam a hipótese de que as florestas com lençol freático raso, historicamente negligenciadas, sejam resilientes à seca moderada, desafiando a visão predominante dos efeitos negativos generalizados da mudança climática nas florestas amazônicas que ignora gradientes de profundidade do lençol freático, mas prevê que elas podem entrar em colapso sob secas muito fortes.

La función de los bosques tropicales es de importancia mundial para las respuestas al cambio climático y está críticamente determinada por los patrones de disponibilidad de agua. El agua subterránea está estrechamente relacionada con el agua del suelo a través de la profundidad del nivel freático (NF), pero históricamente se há negligenciado en los estudios ecológicos. Los bosques con NF poco profundos (NF < 5 m) están subrepresentados en las redes de investigación forestal y ausentes en las mediciones de flujo de gases, aunque representan ~ 50% de la Amazonía y se espera que respondan de manera diferente a las sequías relacionadas con el cambio climático global. Aquí revisamos los patrones de NF y las consecuencias para las plantas, los resultados emergentes y avanzamos en un modelo conceptual que integra distribuciones ambientales y de rasgos funcionales para predecir los efectos del cambio climático. Los bosques con NF poco profundos tienen una composición de especies distinta, con árboles más adquisitivos en la obtención de recursos e hidrológicamente más vulnerables, dosel más bajo y menor biomasa que los bosques de NF profundo. Durante los años climáticos 'normales', los bosques con NF poco profundos tienen una mayor mortalidad y menor productividad que los bosques con NF profundos, pero durante sequías moderadas la mortalidad se amortigua y la productividad aumenta. Sin embargo, durante una sequía severa, los bosques de NF poco profundos pueden ser más sensibles debido a raíces poco profundas y rasgos de intolerancia a la sequía. Nuestra evidencia apoya la hipótesis de que los bosques de NF poco profundos, mayoritariamente desconsiderados, son resistentes a sequías moderadas, desafiando la visión predominante de impactos negativos generalizados del cambio climático en los bosques amazónicos, que ignora los gradientes de NF, pero predice que podrían colapsar bajo sequías muy fuertes.

Giants of the Amazon: How does environmental variation drive the diversity patterns of large trees?

For more than three decades, major efforts in sampling and analyzing tree diversity in South America have focused almost exclusively on trees with stems of at least 10 and 2.5 cm diameter, showing highest species diversity in the wetter western and northern Amazon forests. By contrast, little attention has been paid to patterns and drivers of diversity in the largest canopy and emergent trees, which is surprising given these have dominant ecological functions. Here, we use a machine learning approach to quantify the importance of environmental factors and apply it to generate spatial predictions of the species diversity of all trees (dbh ≥ 10 cm) and for very large trees (dbh ≥ 70 cm) using data from 243 forest plots (108,450 trees and 2832 species) distributed across different forest types and biogeographic regions of the Brazilian Amazon. The diversity of large trees and of all trees was significantly associated with three environmental factors, but in contrasting ways across regions and forest types. Environmental variables associated with disturbances, for example, the lightning flash rate and wind speed, as well as the fraction of photosynthetically active radiation, tend to govern the diversity of large trees. Upland rainforests in the Guiana Shield and Roraima regions had a high diversity of large trees. By contrast, variables associated with resources tend to govern tree diversity in general. Places such as the province of Imeri and the northern portion of the province of Madeira stand out for their high diversity of species in general. Climatic and topographic stability and functional adaptation mechanisms promote ideal conditions for species diversity. Finally, we mapped general patterns of tree species diversity in the Brazilian Amazon, which differ substantially depending on size class.

Metagenomics reveals an abundance of black yeast-like fungi in the skin microbiome.

BACKGROUND: The skin is the first line of defence against communities of resident viruses, bacteria and fungi. The composition of the microbiome might change with factors related to the environment and host. The microbiome is dominated by bacteria. Dermatophytes and yeasts are the predominant fungi that are also involved in opportunistic infections of skin, hair and nails. Among environmental fungi, Chaetothyriales (black yeasts and relatives) are enriched by hydrocarbon pollution in domesticated habitats and comprise numerous species that cause mild-to-severe disease. METHODS: We investigated the presence of black fungi in the skin microbiome by conducting an analysis in the publicly available metagenomic SRA database (NCBI). We focused on the causative agents of chromoblastomycosis and phaeohyphomycosis and used barcodes and padlock probe sequences as diagnostic tools. RESULTS: A total of 132,159,577 MB was analysed and yielded 18,360 reads that matched with 24 species of black fungi. Exophiala was the most prevalent genus, and Cyphellophora europaea was the most abundant species. CONCLUSION: This study reveals the abundant presence of Chaetothyriales on the skin without necessarily being associated with infection. Most of the detected causal agents are known from mild skin diseases, while also species were revealed that had been reported from CARD9-deficient patients.

Isometric tongue endurance and incomplete laryngeal vestibule closure in Parkinson's disease.

BACKGROUND: Dysphagia is a common symptom of Parkinson's disease (PD). A delay in laryngeal vestibule closure (LVC) and a reduction in tongue pressure, may affect swallowing safety and increase the risk of pulmonary aspiration. OBJECTIVE: To verify the relationship between tongue pressure and airway protection in PD patients: (1) comparing tongue pressure measures and physiological events in the pharyngeal phase of swallowing between PD and controls and (2) analysing the association between tongue pressure and LVC in the PD group. METHODS: Twenty-three patients with idiopathic PD (64.9 years) and 24 healthy controls (64.1 years) participated in this study. All participants underwent the following procedures to verify tongue pressure measurements using the Iowa Oral Performance Instrument: maximum anterior and posterior pressure, isotonic and isometric tongue endurance and anterior and posterior tongue pressure during saliva swallowing. To verify swallowing safety, videofluoroscopic swallowing studies focusing on the pharyngeal phase were performed based on the MBSImP protocol. RESULTS: Compared to healthy controls, PD exhibited a statistically significant decline in tongue pressure tasks: posterior maximum pressure, isotonic endurance, anterior and posterior isometric endurance and tongue pressure during posterior swallowing. Patients with PD had worse pharyngeal scores, including LVC scores, than controls. PD and incomplete LVC had lower anterior isometric endurance scores when compared to those with complete LVC. CONCLUSION: PD with incomplete LVC scored lower in the anterior isometric endurance task. We observed a potential clinical use of this task for the assessment and management of dysphagia in patients with PD.

Prevalence of sarcopenia in Parkinson's disease: A systematic review and meta-analysis.

We meta-analysed the sarcopenia prevalence among patients with Parkinson´s disease (PD) in comparison to a control group and tested the effects of age, sex, sarcopenia assessments, and PD progression in the sarcopenia prevalence. The literature search was performed using five databases in March 2022. The prevalence of sarcopenia in patients with PD was 3 times higher than in the control group (OR 3.98). Subgroup analyses showed that among individuals aged ≥ 70 years the higher prevalence of sarcopenia in PD compared to controls (OR 5.32, P=0.08) tended to be higher (P=0.08) than the group < 70 years. Regarding PD progression, the prevalence of sarcopenia was not different between individuals scoring < 2.5 and ≥ 2.5 in the Hoehn and Yahr scale. Patients with PD have a higher probability of developing sarcopenia when compared with the control group and older PD patients trended to have even higher chance of sarcopenia than their older controls.

Osteoid Osteoma Mimicking Arthritis.

ABSTRACT: Intra-articular osteoid osteoma may simulate arthritis, due to the intra-articular presence of prostaglandin, which leads to synovitis, joint effusion, pain, and high local temperature. Also, intra-articular osteoid osteoma may present with minimum or no cortical thickening. Therefore, a high suspicion is needed for a correct and early diagnosis. Perfusion weighted imaging, such as dynamic contrast-enhanced imaging, can aid in the localization of the tumor nidus, seen as an early arterial-phase focal enhancement after the gadolinium injection, with fast washout, as a result of its hypervascularity.

Regulation of l- and d-Aspartate Transport and Metabolism in Acinetobacter baylyi ADP1.

The regulated uptake and consumption of d-amino acids by bacteria remain largely unexplored, despite the physiological importance of these compounds. Unlike other characterized bacteria, such as Escherichia coli, which utilizes only l-Asp, Acinetobacter baylyi ADP1 can consume both d-Asp and l-Asp as the sole carbon or nitrogen source. As described here, two LysR-type transcriptional regulators (LTTRs), DarR and AalR, control d- and l-Asp metabolism in strain ADP1. Heterologous expression of A. baylyi proteins enabled E. coli to use d-Asp as the carbon source when either of two transporters (AspT or AspY) and a racemase (RacD) were coexpressed. A third transporter, designated AspS, was also discovered to transport Asp in ADP1. DarR and/or AalR controlled the transcription of aspT, aspY, racD, and aspA (which encodes aspartate ammonia lyase). Conserved residues in the N-terminal DNA-binding domains of both regulators likely enable them to recognize the same DNA consensus sequence (ATGC-N7-GCAT) in several operator-promoter regions. In strains lacking AalR, suppressor mutations revealed a role for the ClpAP protease in Asp metabolism. In the absence of the ClpA component of this protease, DarR can compensate for the loss of AalR. ADP1 consumed l- and d-Asn and l-Glu, but not d-Glu, as the sole carbon or nitrogen source using interrelated pathways. IMPORTANCE A regulatory scheme was revealed in which AalR responds to l-Asp and DarR responds to d-Asp, a molecule with critical signaling functions in many organisms. The RacD-mediated interconversion of these isomers causes overlap in transcriptional control in A. baylyi. Our studies improve understanding of transport and regulation and lay the foundation for determining how regulators distinguish l- and d-enantiomers. These studies are relevant for biotechnology applications, and they highlight the importance of d-amino acids as natural bacterial growth substrates.

Local hydrological gradients structure high intraspecific variability in plant hydraulic traits in two dominant central Amazonian tree species.

Addressing the intraspecific variability of functional traits helps understand how climate change might influence the distribution of organismal traits across environments, but this is notably understudied in the Amazon, especially for plant hydraulic traits commonly used to project drought responses. We quantified the intraspecific trait variability of leaf mass per area, wood density, and xylem embolism resistance for two dominant central Amazonian tree species, along gradients of water and light availability, while accounting for tree age and height. Intraspecific variability in hydraulic traits was high, with within-species variability comparable to the whole-community variation. Hydraulic trait variation was modulated mostly by the hydrological environment, with higher embolism resistance of trees growing on deep-water-table plateaus compared with shallow-water-table valleys. Intraspecific variability of leaf mass per area and wood density was mostly modulated by intrinsic factors and light. The different environmental and intrinsic drivers of variation among and within individuals lead to an uncoupled coordination among carbon acquisition/conservation and water-use traits. Our findings suggest multivariate ecological strategies driving tropical tree distributions even within species, and reflect differential within-population sensitivities along environmental gradients. Therefore, intraspecific trait variability must be considered for accurate predictions of the responses of tropical forests to climate change.

"Million dollar nerve" magnetic resonance neurography: first normal and pathological findings.

OBJECTIVES: To evaluate prospectively the feasibility of magnetic resonance neurography (MRN) in identifying the anatomical characteristics of thenar muscular branch (TMB) of the median nerve, also known as the "million dollar nerve," in patients and controls. METHODS: Thirteen patients affected by carpal tunnel syndrome (CTS) and four healthy controls had their hands scanned on a 3-T MR imaging scanner for TMB visualization. Median nerve anatomical variations were classified into four groups according to Poisel's classification system modified by Lanz. TMB signal intensity and diameter were assessed for the diagnosis of neuropathy. RESULTS: TMB was successfully identified in all patients and subjects by using MRN. The most suitable pulse sequences to identify and measure nerve diameter were 3D DW-PSIF and T2-FS-TSE. The axial oblique and sagittal oblique planes are complementary in demonstrating its entire course. TMB had mostly an extraligamentous course with radial side origin (93.8%, each). All patients experienced increased T2 signal intensity (p < 0.001) and thickened nerves. Mean TMB diameters were 1.27 ± 0.21 mm (range, 1.02-1.74 mm) and 0.87 ± 0.16 mm (0.73-1.08 mm) (p = 0.008) in the patient and control groups, respectively. CONCLUSION: MRN is a reliable imaging technique for identification and anatomical characterization of TMB in patients affected by CTS. This innovative imaging workup may therefore be included in the preoperative evaluation of patients scheduled for carpal tunnel release, especially in CTS with TMB involvement or even in isolated TMB neuropathy. KEY POINTS: • Magnetic resonance neurography allows precise visualization of the thenar muscular branch of the median nerve. • Thenar muscular branch anatomical variations can be correctly identified. • Preoperative scanning can contribute to reducing the risk of iatrogenic injuries during carpal tunnel release, especially in carpal tunnel syndrome with thenar muscular branch involvement or even in isolated thenar muscular branch neuropathy.

Effects of pharmaceuticals compounds and calcium on granulation, microbiology, and performance of anaerobic granular sludge systems.

Granular sludge is a promising biotechnology to treat sewage contaminated with pharmaceuticals due to its increased toxicity resistance. In this context, this study evaluated the potential of Ca2+ as a granulation precursor and how pharmaceutical compounds (loratadine, prednisone, fluconazole, fenofibrate, betamethasone, 17α-ethinyl estradiol, and ketoprofen) affect granulation. Continuous and intermittent dosages of Ca2+ in the presence and absence of pharmaceuticals were evaluated. The results showed that intermittent addition of Ca2+ reduces the time for anaerobic sludge granulation, and pharmaceuticals presence did not impair granulation. 10% of the granules presented mean diameters greater than 2.11 mm within 93 days with intermittent Ca2+ dosage in the pharmaceuticals' presence. In contrast, no granules higher than 2.0 mm were observed with no precursor addition. The pharmaceuticals' toxicity may have created a stress condition for the microbial community, contributing to more EPS production and a greater potential for granulation. It was also verified that pharmaceuticals' presence did not decrease organic matter, total alkalinity, and volatile fatty acids removals. The 16S rRNA gene analysis revealed taxa resistance to recalcitrant compounds when pharmaceuticals were added. Besides, the efficiency of a granular sludge bioreactor (EGSB) was evaluated for pharmaceuticals removal, and betamethasone, fenofibrate, and prednisone were effectively removed.

The other side of droughts: wet extremes and topography as buffers of negative drought effects in an Amazonian forest.

There is a consensus about negative impacts of droughts in Amazonia. Yet, extreme wet episodes, which are becoming as severe and frequent as droughts, are overlooked and their impacts remain poorly understood. Moreover, drought reports are mostly based on forests over a deep water table (DWT), which may be particularly sensitive to dry conditions. Based on demographic responses of 30 abundant tree species over the past two decades, in this study we analyzed the impacts of severe droughts but also of concurrent extreme wet periods, and how topographic affiliation (to shallow - SWTs - or deep - DWTs - water tables), together with species functional traits, mediated climate effects on trees. Dry and wet extremes decreased growth and increased tree mortality, but interactions of these climatic anomalies had the highest and most positive impact, mitigating the simple negative effects. Despite being more drought-tolerant, species in DWT forests were more negatively affected than hydraulically vulnerable species in SWT forests. Interaction of wet-dry extremes and SWT depth modulated tree responses to climate, providing buffers to droughts in Amazonia. As extreme wet periods are projected to increase and at least 36% of the Amazon comprises SWT forests, our results highlight the importance of considering these factors in order to improve our knowledge about forest resilience to climate change.

Mutational and Functional Analyses of Substrate Binding and Catalysis of the Listeria monocytogenes EutT ATP:Co(I)rrinoid Adenosyltransferase.

ATP:Co(I)rrinoid adenosyltransferases (ACATs) catalyze the transfer of the adenosyl moiety from co-substrate ATP to a corrinoid substrate. ACATs are grouped into three families, namely, CobA, PduO, and EutT. The EutT family of enzymes is further divided into two classes, depending on whether they require a divalent metal ion for activity (class I and class II). To date, a structure has not been elucidated for either class of the EutT family of ACATs. In this work, results of bioinformatics analyses revealed several conserved residues between the C-terminus of EutT homologues and the structurally characterized Lactobacillus reuteri PduO (LrPduO) homologue. In LrPduO, these residues are associated with ATP binding and formation of an intersubunit salt bridge. These residues were substituted, and in vivo and in vitro data support the conclusion that the equivalent residues in the metal-free (i.e., class II) Listeria monocytogenes EutT (LmEutT) enzyme affect ATP binding. Results of in vivo and in vitro analyses of LmEutT variants with substitutions at phenylalanine and tryptophan residues revealed that replacement of the phenylalanine residue at position 72 affected access to the substrate-binding site and replacement of a tryptophan residue at position 238 affected binding of the Cbl substrate to the active site. Unlike the PduO family of ACATs, a single phenylalanine residue is not responsible for displacement of the α-ligand. Together, these data suggest that while EutT enzymes share a conserved ATP-binding motif and an intersubunit salt bridge with PduO family ACATs, class II EutT family ACATs utilize an unidentified mechanism for Cbl lower-ligand displacement and reduction that is different from that of PduO and CobA family ACATs.

Embolism resistance drives the distribution of Amazonian rainforest tree species along hydro-topographic gradients.

Species distribution is strongly driven by local and global gradients in water availability but the underlying mechanisms are not clear. Vulnerability to xylem embolism (P50 ) is a key trait that indicates how species cope with drought and might explain plant distribution patterns across environmental gradients. Here we address its role on species sorting along a hydro-topographical gradient in a central Amazonian rainforest and examine its variance at the community scale. We measured P50 for 28 tree species, soil properties and estimated the hydrological niche of each species using an indicator of distance to the water table (HAND). We found a large hydraulic diversity, covering as much as 44% of the global angiosperm variation in P50 . We show that P50 : contributes to species segregation across a hydro-topographic gradient in the Amazon, and thus to species coexistence; is the result of repeated evolutionary adaptation within closely related taxa; is associated with species tolerance to P-poor soils, suggesting the evolution of a stress-tolerance syndrome to nutrients and drought; and is higher for trees in the valleys than uplands. The large observed hydraulic diversity and its association with topography has important implications for modelling and predicting forest and species resilience to climate change.