Pleurotus pulmonarius: a protease-producing white rot fungus in lignocellulosic residues.

The production of proteases by white rot fungi, such as those of the genus Pleurotus, is related to the degradation of wood proteins, the substrate on which these fungi grow in the environment. From the point of view of production, they are still little explored for this purpose. A selection of agro-industrial residues highlighted corn bagasse as the best substrate for solid-state protease production using the basidiomycete Pleurotus pulmonarius. The enzyme production was maximized through a factorial design, where the enzyme activity increased from 137.8 ± 1.9 to 234.1 ± 2.7 U/mL. Factors such as temperature stability, pH, and chemical reagents were evaluated. The optimum temperature was 45 °C, showing low thermal stability at higher temperatures. The enzyme inhibition occurred by Mn2+ (50.3%) and Ba2+ (76.4%); SDS strongly inhibited the activity (82.4%), while pepstatin A partially inhibited (56%), suggesting an aspartic protease character. Regarding pH, the highest protease activity was obtained at pH 5.5. Partial characterization resulted in apparent values of the KM and Vmax constants of 0.61 mg/mL and 1.79 mM/min, respectively.

Teaching and learning pharmacology in Brazil before COVID-19 pandemic: a case study in Rio de Janeiro.

BACKGROUND: Knowledge of pharmacology is crucial for physicians to perform rational and safe medicine. Medical professionals are responsible for prescribing drugs and a weak performace of those can result in medication errors leading to disability, hospitalization, and death, among other situations. It occurs worldwide, including in Brazil, so that learning pharmacology impacts on public health service. We aim to investigate the current pharmacology educational practices in medical schools in the state of Rio de Janeiro, Brazil. METHODS: We surveyed 14 of 22 medical schools in Rio de Janeiro. Pharmacology teachers (n=16) and medical students (n=89) answered a semi-structured questionnaire that included questions about the staff characteristics, pharmacology content, teacher's concepts, and common practices and resources that were used in pharmacology classes. RESULTS: Our results revealed that the medical schools had similar overall curriculums. Pharmacology teachers work more than 30hs a week (75%) and conducted both research and teaching (62.5%). We also found that the multimedia projector was the most common resource (71.9%), and passive pedagogical methodologies (e.g., expository classes) remain a current strategy in pharmacology classes (89.9%). In general, medical students are poorly motivated (55%), which may be related to their performance in assessments. In addition, students believe that pharmacology is a complex (52%) or very complex subject (46%) since for its full understanding the student needs concepts from other disciplines, which can have an impact on the performance and motivation of students. As a result, these medical students do not fully understand the integration between pharmacology's basic concepts and their clinical applications. CONCLUSION: These data seem to demonstrate that the adopted teaching and learning pharmacology strategies and methodologies can be improved in Rio de Janeiro.

Influence of Iron on Physiological Parameters and Intracellular Microcystin in Microcystis Panniformis Strain Isolated from a Reservoir in the Amazon.

In the Amazon, the leaching from soil left unprotected by deforestation increases the entry of iron, among other elements, in aquatic ecosystems, which can cause cyanobacterial blooms. This study aimed to investigate the physiological response of a strain of Microcystis panniformis to iron variation. The strain was isolated from a reservoir located in the Western Amazon and produces microcystin-LR. After a period of iron deprivation, the cultures were submitted to three conditions: control (223 µgFe.L-1), treatment with 23 µgFe.L-1, and absence of iron. At regular intervals for eight days, the cell density, levels of chlorophyll a and microcystins were determined. On the second and fourth day, transcription of genes responsive to iron limitation was quantified. Starting on the fourth day of the experiment, the different iron concentrations affected growth, and on the eighth day in the iron-free condition cell density was 90% lower than in control. Chlorophyll cell quota in 23 µgFe.L-1 and control presented similar values, while without iron the cells became chlorotic as of the fourth day Toxin concentration in cells grow in 0 µgFe.L-1 in relation to the control. Higher transcription levels of the feo and fut genes were observed in the 0 µgFe.L-1 and 23 µgFe.L-1 treatments, indicating that the cells were activating high-affinity capture systems to reestablish an adequate concentration of intracellular iron. The increasing deforestation in the Jamari River Basin (Amazon region), can contribute to the occurrence of toxic cyanobacterial blooms due to the greater entrance of iron in water bodies.

Mercury biomagnification in an ichthyic food chain of an amazon floodplain lake (Puruzinho Lake): Influence of seasonality and food chain modeling.

Mercury (Hg) biomagnification in fish food chains is a relevant subject due to the high fish consumption of the Amazonian population and the high toxicity of this metal. In the Amazon, floodplain lake hydrodynamics change considerably along the four seasons of the hydrological cycle (rising water, high water, falling water and low water), which can influence Hg bioaccumulation in fish. The main aim of this study was to evaluate if Hg biomagnification is influenced by seasonality in a floodplain lake (Puruzinho Lake) in the Brazilian Amazon. Additionally, the influence of food chain modeling on measurement of Hg biomagnification was tested. Hg concentrations and stable isotope signatures (carbon and nitrogen) were estimated in four species, Mylossoma duriventre (herbivorous), Prochilodus nigricans (detritivorous), Cichla pleiozona (piscivorous) and Serrasalmus rhombeus (piscivorous). The "trophic magnification slope" (TMS) of the food chain composed by the four species was calculated and compared among the four seasons. There was no significant seasonal variation in TMS among rising water, high water, falling water and low water seasons (p = 0.08), suggesting that Hg biomagnification does not change seasonally. However, there was significant variation in TMS among different food chain models. Lower TMS was observed in a food chain composed of detritivorous and piscivorous fish (0.20) in comparison with a food chain composed of the four species (0.26). The results indicate food chain modeling influences TMS results.

Use of an evidence-based health portal to improve teaching and learning in primary care: a mixed methods evaluation.

BACKGROUND: Worldwide, primary care is for most people the gateway into many health systems. Offering solutions to the demands of the communities served requires the constant preparation of professionals, especially doctors and medical undergraduate students. We analyze and propose ways to improve the teaching and learning processes facilitated by the Basic Family Health Units (BFHUs) based on the use of electronic portals with evidence-based medicine criteria. METHOD: First phase: The authors conducted a qualitative-quantitative study on students and instructors of primary care (PC) medicine by administering a survey of open- and closed-ended questions at medical schools. The closed-ended questions were studied with descriptive statistics, and open-ended questions were analyzed via the creation of categories. Perceptions of major teaching and learning problems were then identified. Second phase: Meetings were held with students and their instructors for 6 months and involved the use of electronic portals and the application of new questionnaires using a Likert scale for pre- and postevaluation. RESULTS: In the first phase, 40% of the students considered local instructor training levels a problem. A similar result was found regarding teachers' lectures, revealing a lack of adequate PC training and performance. Building on our results, we focused on BFHUs to apply new strategies for teaching and learning, such as the use of the Evidence-Based Health (SBE) Portal, which includes several databases with clinical evidence criteria. In the second phase, the authors identified an improvement in the quality of learning among instructors and students. This outcome improved safety in daily clinical practice in PC, possibly with better results for its users. CONCLUSIONS: The use of electronic portals can facilitate BFHU teaching and learning and promote the health of users.

Arsenic in the lower drainage basin of the Paraíba do Sul River (Southeast Brazil): dynamics between the water column and sediment.

The presence of arsenic (As) in the sediment and the particulate and dissolved fractions of the water column determines its behavior and bioavailability. The main geochemical parameters responsible for As mobility are organic matter and oxide-forming metals such as Fe. The As distribution and its concentration were evaluated in the dissolved fraction, suspended particulate matter (SPM) and sediment of the lower Paraíba do Sul River (PSR), its main tributaries (Muriaé, Pomba, and Dois Rios rivers), and flooded and estuarine areas. As was not detected in the dissolved fraction. The river flow influenced the As concentration in the SPM, which was higher in the dry season than in the rainy season (2.6 ± 0.69 and 1.98 ± 0.29 mg kg-1, respectively). The Fe oxyhydroxides, organic carbon, and surface area measured in the sediment were positively related with As concentration (R2 = 0.11, 0.34, and 0.30; p < 0.05). The highest As concentrations in the sediment occurred in the secondary estuary and flooded areas (5.16 ± 4.78 and 1.23 ± 0.44 mg kg-1, respectively), in which finer granulometric fractions (silt and clay) predominated (64% and 71%, respectively), in addition to greater surface area. The measurement of stable carbon and nitrogen isotopes (δ13C and δ15N) and atomic ratio (C:N)a indicated the existence of a combination of autochthonous and allochthonous sources of organic matter composing the SPM. In general, the As concentrations in the sediment and SPM were low, with values below that permitted by Brazilian regulations (5.9 mg kg-1), which suggests that the As sources in the lower basin of the PSR are diffuse and natural.

Variation in Hg accumulation between demersal and pelagic fish from Puruzinho Lake, Brazilian Amazon.

Aquatic ecosystems in the Amazon are exposed to mercury, mostly from natural sources. Hg accumulation in fish tissues poses a risk to the local population since fish is one of the main sources of protein in the region. The aim of this study was to evaluate Hg distribution in demersal and pelagic carnivorous fish between seasons in Puruzinho Lake in the Brazilian Amazon. Total Hg was quantified in 221 individuals of 8 species obtained during the high water and low water seasons. Two-way ANOVA indicated an interaction between foraging habitat and season. During high water, total Hg concentrations were similar between demersal and pelagic fish, while in low water, total Hg levels were higher in demersal fish. Pelagic and demersal fishes' Hg levels were similar between the two seasons.

Influence of the flood pulse on mercury accumulation in detritivorous, herbivorous and omnivorous fish in Brazilian Amazonia.

Hg accumulation in fish is influenced by several factors including seasonality. In the Amazon, ecosystems are marked by strong seasonal variation in precipitation, which leads to drastic changes in the water level of lakes and rivers. The aim of this study was to evaluate Hg levels in muscle of detritivorous, herbivorous and omnivorous fish from an Amazon lake (Madeira River Basin, Amazonas, Brazil) over four seasons (rising water, high water, falling water and low water). We hypothesized that total Hg concentration varies during the seasons. The results indicate that total Hg levels in detritivorous fish were higher in rising and low water seasons while in herbivorous and omnivorous fish the total Hg concentration was higher during the rising water season. The hypothesis was supported by the results. Additionally, the study provides evidence that Hg levels in fish with different feeding habits are influenced by the flood pulse of the Amazon region.

Arsenic Exposure and Effects in Humans: A Mini-Review in Brazil.

Arsenic (As) is widely studied in several countries due to its toxicity to biota in the environment. Arsenic sources may be natural or anthropogenic, and the mobility of the element is ruled by physicochemical conditions that also define the dominant As species in the environment. Arsenic levels are evaluated in various abiotic and biotic environmental samples. The highest As levels are observed in sediment, from where it may be mobilized into the aquifers. This article reviews studies about As in the world but with emphasis on studies performed in Brazil, a country where continental water bodies are a common geographic feature. We reviewed 64 studies published between 1985 and 2016. The results indicate that in recent years more studies have been conducted to determine As levels in foods and human samples as a tool to evaluate the exposure of populations and identify potential sources. In Brazil, the main problems associated with contamination with As are the use of wood preservatives and herbicides as well as the impact caused by mining. Also, the precarious character of sewage treatment systems contributes to the contamination of water bodies.

Exposure to toxic and essential trace elements through the intake of processed and meat cuts (beef and chicken) in southeastern Brazil.

The present study evaluated the concentration of six trace elements in processed meat products and in meat cuts. We also assessed the risk associated with the consumption of these foods based on the estimated daily intake (EDI) of these elements. Fifty-eight processed meat and 148 meat cuts samples were analyzed using ICP-OES. As and Cd were not detected in any sample of processed meat. The highest mean level of lead was observed in frankfurters (0.056 µg g-1), which is half the maximum permissible level. For Cr, the highest mean concentrations were detected in chicken nuggets and beef hamburger (0.121 and 0.105 µg g-1, respectively), which are above the allowed limit. The comparison between the impact of a diet restricted to processed meats with a diet restricted to meats cuts showed that the individuals following the latter are exposed to higher amounts of the trace elements analyzed.

Bioconcentration and translocation of Cd and Hg in a tomato (Solanum lycopersicum) from cultivated soils in southeastern Brazil.

Food is the main source of toxic metals like mercury (Hg) and cadmium (Cd) to humans. This study evaluated the accumulation and translocation of Cd and Hg in a soil-plant system in four tomato plantations and assessed the risk associated with ingestion of these metals. Ten soil samples (rhizosphere) and 10 samples of plant tissues (leaf, ripe fruit, green fruit, and roots) were collected in each plantation. Spatial variation in Cd and Hg concentrations was negligible. The Cd level in rhizosphere was lower in comparison with that of plant tissues. Hg levels in rhizosphere were similar to roots and higher than the value observed in aerial parts of plants. The Cd bioconcentration factor was approximately five times higher compared to that of Hg. Approximately 93% and 48.6% of Cd and Hg accumulated in roots reached aerial parts, respectively. Our results indicate that tomato readily absorbs Cd accumulated in soil, translocating it to aerial parts. Comparatively, the absorption of Hg is not efficient. Levels of Cd in tomatoes were over 17 times higher than the maximum residual levels in 57.5% of ripe fruits and in 27.5% of green tomatoes.

Evaluation of antitumor activity and cardiac toxicity of a bone-targeted ph-sensitive liposomal formulation in a bone metastasis tumor model in mice.

Chemotherapy for bone tumors is a major challenge because of the inability of therapeutics to penetrate dense bone mineral. We hypothesize that a nanostructured formulation with high affinity for bone could deliver drug to the tumor while minimizing off-target toxicity. Here, we evaluated the efficacy and toxicity of a novel bone-targeted, pH-sensitive liposomal formulation containing doxorubicin in an animal model of bone metastasis. Biodistribution studies with the liposome showed good uptake in tumor, but low accumulation of doxorubicin in the heart. Mice treated with the bone-targeted liposome formulation showed a 70% reduction in tumor volume, compared to 35% reduction for free doxorubicin at the same dose. Both cardiac toxicity and overall mortality were significantly lower for animals treated with the bone-targeted liposomes compared to free drug. Bone-targeted, pH-sensitive, doxorubicin containing liposomes represent a promising approach to selectively delivering doxorubicin to bone tumors while minimizing cardiac toxicity.

Impact of Land Use on the Mobility of Hg Species in Different Compartments of a Tropical Watershed in Brazil.

This study evaluated the levels of total Hg and CH3Hg+ from a comprehensive perspective, considering the retention, leaching, and deposition of these contaminants in the main compartments (soil, plant litter, and sediment) of three landscapes (Atlantic Forest, pasture, and agricultural area) in a watershed in northern Rio de Janeiro State, Brazil. Variables analyzed were total Hg, CH3Hg+, organic carbon, total nitrogen, grain size, and surface area. In soil samples, total Hg levels were the highest in agricultural soil followed by forest soil and pasture (97.3, 87.6, and 77.1 ng g-1, respectively), and CH3Hg+ was lower than 1.7%. Total Hg levels in leaf litter varied between 22.6 and 34.2 ng g-1, and CH3Hg+ was 4.37%. In sediment, Hg (60-180 ng g-1) and CH3Hg+ (<1%) indicate the transport of these contaminants from soils to this compartment and may be associated with soil use and cover. Multiple regressions were used to understand the dispersion of Hg species, and the effect of each variable varied with the landscape, showing that plant cover should not be ignored in investigations related to Hg species retention in a watershed. The landscapes surveyed in the present study clearly influence the quantitative and qualitative distribution of Hg species. On the other hand, anthropic processes associated with changes in soil use did not have any critical effects on the absolute levels of total Hg and CH3Hg+, meaning that the landscapes evaluated seem to represent the background concentration of these chemical species for the evaluated watershed.

The P2X7 receptor: shifting from a low- to a high-conductance channel - an enigmatic phenomenon?

The general structure of the P2X7 receptor (P2X7R) is similar to the structure of other P2X receptor family members, with the exception of its C terminus, which is the longest of this family. The P2X7R activates several intracellular signaling cascades, such as the calmodulin, mitogen-activated protein kinase and phospholipase D pathways. At low concentrations of ATP (micromolar range), P2X7R activation opens a cationic channel, similarly to other P2X receptors. However, in the presence of high concentrations of ATP (millimolar range), it opens a pathway that allows the passage of larger organic cations and anions. Here, we discuss both the structural characteristics of P2X7R related to its remarkable functions and the proposed mechanisms, including the dilation of the endogenous pore and the integration of another channel. In addition, we highlight the importance of P2X7R as a therapeutic target.

Canine low-grade intra-orbital myxosarcoma: case report.

OBJECTIVE: The aim of this study was to evaluate important clinical, morphological, histopathological, histochemical, and immunohistochemical characteristics in order to establish the diagnosis and prognosis of a low-grade intra-orbital myxosarcoma. ANIMAL STUDIED: A mongrel dog presented a 2-year history of a neoplastic mass behind the right eye. RESULTS: The neoplasm presented a mesenchymal spindle and stellate cell proliferation with an abundant myxoid matrix, moderate anisocariosis, and a low mitotic index. It stained positive for vimentin, moderately positive for periodic acid-Schiff, and negative for Gomori trichrome stain and α-smooth muscle actin. CONCLUSIONS: One year following surgical excision, the patient remains disease free. The histological findings established a diagnosis of a rare canine intra-orbital low-grade myxosarcoma.

Evaluation of different methods of cryopreservation of Ehrlich tumor cells.

BACKGROUND: The Ehrlich Ascitic Carcinoma (EAC) is an experimental transplantable neoplasm that develops in several species of mice. The maintenance of the tumor occurs in vivo. Thus, freezing the cells would reduce the number of passages between animals, ensuring genetic stability and storage for longs period of experimentation. OBJECTIVE: Search by EAC cryoprotectants. MATERIALS AND METHODS: The combinations of nutrient medium (Tris, hen egg yolk, and DEMEN) and cryoprotective agent (Glicerol, Trehalose and DMSO) on freezing EAC cells and the transplantability after defrosting were evaluated. The cooling was conducted at 2 C/min. until -180 degree C and the thawing by immersion in water at 37 degree C. The transplantability was evaluated from cell inoculation in mice for 14 days. RESULTS: The best results were the associations IA (Cryoprotective agent Glycerol 6 % and medium containing 3.0 % Tris w / v, 1.8 % Citric acid w / v, 1.3 % D-fructose w / v and 20 % hen egg yolk v / v) and IIB (Cryoprotective agent Trehalose 100mM and medium containing 50 % coconut water v / v, 25 % sodium citrate 5 % v / v and 20 % hen egg yolk v / v) with 85.2 % and 55.1 % viable cells, respectively. CONCLUSION: These transplantable cells were efficient for tumor development, therefore demonstrating that this method of cryopreservation is simple and affordable.

Endophytic fungi: expanding the arsenal of industrial enzyme producers.

Endophytic fungi, mostly belonging to the Ascomycota, are found in the intercellular spaces of the aerial plant parts, particularly in leaf sheaths, sometimes even within the bark and root system without inducing any visual symptoms of their presence. These fungi appear to have a capacity to produce a wide range of enzymes and secondary metabolites exhibiting a variety of biological activities. However, they have been only barely exploited as sources of enzymes of industrial interest. This review emphasizes the suitability and possible advantages of including the endophytic fungi in the screening of new enzyme producing organisms as well as in studies aiming to optimize the production of enzymes through well-known culture processes. Apparently endophytic fungi possess the two types of extracellular enzymatic systems necessary to degrade the vegetal biomass: (1) the hydrolytic system responsible for polysaccharide degradation consisting mainly in xylanases and cellulases; and (2) the unique oxidative ligninolytic system, which degrades lignin and opens phenyl rings, comprises mainly laccases, ligninases and peroxidases. The obvious ability of endophytic fungi to degrade the complex structure of lignocellulose makes them useful in the exploration of the lignocellulosic biomass for the production of fuel ethanol and other value-added commodity chemicals. In addition to this, endophytic fungi may become new sources of industrially useful enzymes such as lipases, amylases and proteases.

Structural and molecular modeling features of P2X receptors.

Currently, adenosine 5'-triphosphate (ATP) is recognized as the extracellular messenger that acts through P2 receptors. P2 receptors are divided into two subtypes: P2Y metabotropic receptors and P2X ionotropic receptors, both of which are found in virtually all mammalian cell types studied. Due to the difficulty in studying membrane protein structures by X-ray crystallography or NMR techniques, there is little information about these structures available in the literature. Two structures of the P2X4 receptor in truncated form have been solved by crystallography. Molecular modeling has proven to be an excellent tool for studying ionotropic receptors. Recently, modeling studies carried out on P2X receptors have advanced our knowledge of the P2X receptor structure-function relationships. This review presents a brief history of ion channel structural studies and shows how modeling approaches can be used to address relevant questions about P2X receptors.

Biotechnological, Nutritional, and Therapeutic Applications of Quinoa (Chenopodium quinoa Willd.) and Its By-Products: A Review of the Past Five-Year Findings.

This study aimed to provide an updated critical review of the nutritional, therapeutic, biotechnological, and environmental aspects involved in the exploitation of Chenopodium quinoa Willd and its biowastes. Special attention was devoted to investigations of the therapeutic and nutritional properties of different parts and varieties of quinoa as well as of the use of the biowaste resulting from the processing of grain. Studies published from 2018 onward were prioritized. Extracts and fractions obtained from several Chenopodium quinoa matrices showed antioxidant, antidiabetic, immunoregulatory, neuroprotective, and antimicrobial effects in in vitro and in vivo models and some clinical studies. The activities were attributed to the presence of phytochemicals such as polyphenols, saponins, peptides, polysaccharides, and dietary fibers. Quinoa wastes are abundant and low-cost sources of bioactive molecules for the development of new drugs, natural antioxidants, preservatives, dyes, emulsifiers, and carriers for food and cosmetics applications. Among the demands to be fulfilled in the coming years are the following: (1) isolation of new bioactive phytochemicals from quinoa varieties that are still underexploited; (2) optimization of green approaches to the sustainable recovery of compounds of industrial interest from quinoa by-products; and (3) well-conducted clinical trials to attest safety and efficacy of extracts and compounds.

A consensus segment in the M2 domain of the hP2X(7) receptor shows ion channel activity in planar lipid bilayers and in biological membranes.

The P2X(7) receptor (P2X(7)R) is an ATP-gated, cation-selective channel permeable to Na(+), K(+) and Ca(2+). This channel has also been associated with the opening of a non-selective pore that allows the flow of large organic ions. However, the biophysical properties of the P2X(7)R have yet to be characterized unequivocally. We investigated a region named ADSEG, which is conserved among all subtypes of P2X receptors (P2XRs). It is located in the M2 domain of hP2X(7)R, which aligns with the H5 signature sequence of potassium channels. We investigated the channel forming ability of ADSEG in artificial planar lipid bilayers and in biological membranes using the cell-attached patch-clamp techniques. ADSEG forms channels, which exhibit a preference for cations. They are voltage independent and show long-term stability in planar lipid bilayers as well as under patch-clamping conditions. The open probability of the ADSEG was similar to that of native P2X(7)R. The conserved part of the M2 domain of P2X(7)R forms ionic channels in planar lipid bilayers and in biological membranes. Its electrophysiological characteristics are similar to those of the whole receptor. Conserved and hydrophobic part of the M2 domain forms ion channels.