Acute murine-betacoronavirus infection impairs testicular steroidogenesis and the quality of sperm production.

Although several testicular alterations promoted by coronavirus infection have been demonstrated, the extent, causes, and players of testicular pathogenesis are not totally understood. The present study aimed to investigate the short-term effects on male fertility of intranasally administered murine hepatitis virus strain 3 (MHV-3), a member of the genus Betacoronavirus, which causes a severe systemic acute infection. This mouse model might be used as a in vivo prototype for investigating the impact of betacoronavirus on the endocrine and exocrine testicular functions with the advantage to be performed in a biosafety level 2 condition. Herein, we performed virological, histopathological, and molecular studies regarding the testicular spermatogenesis and the spermatic quality analyses in an MHV-3-infected C57BL/6 mice. The main outcomes showed that MHV-3 infects mouse testis and induces a testicular inflammatory state, impairing the steroidogenic pathway. The infection led to several alterations in the testicular parenchyma, such as: seminiferous epithelium sloughing, retention of residual bodies, germ cell apoptosis, alterations in intercellular junction proteins, and worse spermatogenic parameters. Moreover, the levels of plasmatic testosterone as well as the quality of sperm production reduced. Therefore, the present data suggest that the viral/inflammatory impairment of the steroidogenic pathway and the consequent imbalance of androgen levels is critical in testicular pathology, disturbing the SC barrier function and the germ cell differentiation. Our study is important for comprehending the effects of beta coronavirus infections on testis function in order to develop treatments that could prevent virus-mediated male infertility.

Coupled hydrodynamic modelling approach to assess land use change induced flood characteristics.

MIKE 11 rainfall-runoff model and MIKE 21 overland flow model were successfully coupled in the MIKE FLOOD platform for flood simulation in Gin catchment (932 km2) of Sri Lanka to assess land use change induced changes in flood discharge. MIKE 11 Nedbør-Afstrømings-Model (NAM) rainfall-runoff simulation depicted a good agreement with the observed discharge at Thawalama and Baddegama gauging stations. MIKE FLOOD, validated against the two major flood events that occurred in May 2003 and in May 2017, showed a reasonable agreement with the observed water depths and peak discharge values displaying more than 70% goodness of fit between the observed and simulated inundated extents. Dominant land use change processes in the catchment between 1999 and 2016 were identified as the forest area and built-up land expansion at the expense of agricultural land and bare land which possessed contradictory impacts on flood generation. The impact of a single factor, 17-year land use change, on flood formation was differentiated. In the upstream sub-catchment having 490 km2, despite a 0.74% increase in the built-up land, 2.85% increase in the forest area had significantly contributed to mitigate the overall flood formation with 34% and 40% reduction in the peak discharge and the flood volume, respectively. Overall reduction of the flood discharge attributed to the forest expansion emphasised the importance of preserving forest cover and pervious area. The modelling framework presented in this typical tropical monsoon catchment study could be effectively used to quantify the land use change induced flow regime variations in similar catchments.

Investigation of the Flexural Behavior of Preloaded and Pre-Cracked Reinforced Concrete Beams Strengthened with CFRP Plates.

This paper investigates the flexural behavior of preloaded reinforced concrete (RC) beams, strengthened with Carbon Fiber Reinforced Polymer (CFRP) plates using an experimental program, analytical procedure, and Finite Element Method (FEM) simulation. The RC beams were subjected to preloads of 30%, 50% and 70% of the yielding load, prior to installation of the strengthening system. The eight RC-strengthened beams with a reinforcement configuration of 3Ø12 and two CarboDur S512 plates have been evaluated using bending tests. The failure modes of all the RC-strengthened beams were governed by the widening of flexural cracks within a constant bending zone, followed by debonding of the CFRP plates. The plates were debonding simultaneously or one plate prior to the other plate. The ultimate moment capacity is not significantly reduced while increasing preload levels from 0% to 70%. The moment capacity is increased by 70% to 80% in the CFRP strengthened beams, compared with un-strengthened beams indicating the potential of capacity enhancement that can be attained by externally bonded CFRP.

Use of in-filled trenches to screen ground vibration due to impact pile driving: experimental and numerical study.

Vibration generated by pile driving can cause discomfort to occupants of nearby buildings and disturb the activities carried out in the buildings. The transmitted vibration will depend on both the source and the transmitting medium (soil), while the acceptable levels of vibration will depend on the receiver characteristics. Existing structures in which some sensitive processes are ongoing such as hospitals and laboratories can easily be affected due to the received vibration. Introducing a trench into the path of wave propagation has become one of the solutions. There is however little experimental data available on the effects of trenches to screen such ground borne vibration, especially that caused by pile driving. This paper describes a series of experiments conducted to investigate the characteristics of impact pile induced vibrations and the effect of coal bottom ash filled trenches to screen this vibration. In addition to experimental testing, numerical simulations are also carried out using validated model to examine the effects of in-fill material, impact load, soil characteristics and distance from the source to the trench on the vibration screening ability. The results of the field experiments and the numerical study are analysed and interpreted to provide guidelines for future research and design.

Prophylactic Bifidobacterium adolescentis ATTCC 15703 supplementation reduces partially allergic airway disease in Balb/c but not in C57BL/6 mice.

Allergic asthma is a chronic disease mainly characterised by eosinophil inflammation and airway remodelling. Many studies have shown that the gut microbiota of allergic individuals differs from that of non-allergic individuals. Although high levels of bifidobacteria have been associated with healthy persons, Bifidobacterium adolescentis ATCC 15703, a gut bacteria, has been associated with allergic individuals in some clinical studies. The relationship between B. adolescentis ATCC 15703 and asthma or allergies has not been well elucidated, and its effect may be dependent on the host's genetic profile or disease state. To elucidate this question, we evaluated the role of preventive B. adolescentis ATCC 15703 treatment on experimental allergic airway inflammation in two genetically different mouse strains, Balb/c and C57BL/6 (B6). Balb/c mice display a greater predisposition to develop allergic responses than B6 mice. Oral preventive treatment with B. adolescentis ATCC 15703 modulated experimental allergic airway inflammation, specifically in Balb/c mice, which showed decreased levels of eosinophils in the airway. B6 mice did not exhibit any significant alterations in eosinophils but showed an increased influx of total leukocytes and neutrophils into the airway. The mechanism underlying the beneficial effects of these bacteria in experimental allergic mice may involve products of bacteria metabolism, as dead bacteria did not mimic the ability of live B. adolescentis ATCC 15703 to attenuate the influx of eosinophils into the airway. To conclude, preventive oral B. adolescentis ATCC 15703 treatment can attenuate the major characteristic of allergic asthma, eosinophil airway influx, in Balb/c but not B6 mice. These results suggest that oral treatment with this specific live bacterial strain may have therapeutic potential for the treatment of allergic airway disease, although its effect is mouse-strain-dependent.

Physiologic and metabolic effects of exogenous kojic acid and tyrosol, chemicals produced by endophytic fungus, on wheat seeds germination.

Antioxidant compounds have the ability to scavenge the reactive oxygen species in an attempt to minimise damage in seeds during the germination. This study aimed to evaluate the physiological and metabolic process of two well-established antioxidant compounds: kojic acid and hydroxyphenyl ethanol (tyrosol), at increasing concentrations, on wheat seeds. The use of different concentrations of tyrosol or kojic acid not showed any interference on seed germination rate. However, we observed isolated effect of antioxidants and their concentrations to: germination speed index, shoot length and electrolyte leakage; and significant interaction between the factors to: seedling total length, seedling fresh matter and α-amylase activity. Our results suggest that the use of antioxidant molecules can be applied on seed treatments for protection against damage oxidative stress and improve seed metabolism.

Metal fractionation in sludge from sewage UASB treatment.

This study evaluates the trace metal composition and fractionation in sludge samples from anaerobic sewage treatment plants from six cities in Brazil. Ten metals were evaluated: Ni, Mn, Se, Co, Fe, Zn, K, Cu, Pb and Cr. Specific methanogenic activity of the sludge was also evaluated using acetic acid as the substrate. Among the essential trace metals for anaerobic digestion, Se, Zn, Ni and Fe were found at a high percentage in the organic matter/sulfide fraction in all sludge samples analyzed. These metals are less available for microorganisms than other metals, i.e., Co and K, which were present in significant amounts in the exchangeable and carbonate fractions. Cu is not typically reported as an essential metal but as a possible inhibitor. One of the samples showed a total Cu concentration close to the maximal amount allowed for reuse as fertilizer. Among the non-essential trace metals, Pb was present in all sludge samples at similar low concentrations and was primarily present in the residual fraction, demonstrating very low availability. Cr was found at low concentrations in all sludge samples, except for the sludge from STP5; interestingly, this sludge presented the lowest specific methanogenic activity, indicating possible Cr toxicity.

Resistance training suppresses intra-abdominal fatty acid synthesis in ovariectomized rats.

Ovarian hormone loss is associated with a shift in fat distribution to intra-abdomin al adipose tissue (intra-AAT) depots and with lipid metabolism disorders, which predisposes individuals to developing insulin resistance. Resistance training (RT) prevents increases in intra-AAT after ovarian hormone loss. However, the molecular mechanisms underlying these changes remain unclear. We investigated the effects of ovariectomy and RT on gene expression related to lipogenesis and fat oxidation in the intra-AAT of ovariectomized rats. Sprague-Dawley rats (n=6/group) were divided into the groups: sham-sedentary, ovariectomized-sedentary, sham-RT and ovariectomized-RT. RT groups performed a 10-week climbing program on a ladder with progressive overload. Intra-AAT was subjected to morphometric and mRNA analysis. Ovariectomized-sedentary group had larger adipocytes and higher expression of peroxisome proliferator-activated receptor-γ (PPAR-γ), sterol regulatory element-binding protein-1c (SREBP-1c), stearoyl-CoA desaturase-1 (SCD-1), acetyl-CoA carboxylase (ACC), hormone-sensitive lipase (HSL) and lower expression of the oxidative carnitinepalmitoyltransferase-I (CPT-1). RT counteracted OVX-induced increases in PPAR-γ and SCD-1 and decreased SREBP-1c. ACC and HSL were downregulated in ovariectomized-RT compared with the ovariectomized-sedentary group. Ovariectomized-RT group had the highest CPT-1 gene expression. Adipocyte size decreased in ovariectomized-RT group. Results suggest that RT reduces intra-AAT adipocyte size in ovariectomized rats by suppressing intra-AAT fatty acid synthesis and enhancing fatty acid ß-oxidation.

Avaliação da atividade hepatoprotetora do asiaticosídeo em modelo experimental de lesão hepática por paracetamol em ratos / Hepatoprotective activity of asiaticoside in experimental liver injury induced by paracetamol in rats

Asiaticoside is a triterpenoid present in Centella asiatica extract, responsible for the therapeutic activity of this plant in chronic liver disease. The hepatocyte is the cell responsible for the endocrine and exocrine functions of the liver, in addition to the conversion of harmful substances into non-toxic compounds that are excreted in the bile. That is why the liver is sensitive to the action of some drugs, such as paracetamol. Hence, paracetamol was used as an experimental model of liver damage, with the aim of assessing the effectiveness of asiaticoside, in a standard therapeutic dose, as a hepatoprotector in Wistar rats. In this experiment, 40 animals were used and divided into two groups: those treated with asiaticoside and the untreated control group. Animals from the first group were subjected to pretreatment with the active ingredient (1mg/kg/dia P.O.) for eight days and exposed to a toxic dose of paracetamol (3 g/kg P.O.) on the eighth day. After 24 h and 72 h, these rats were sacrificed for the collection of blood samples and liver fragments. To assess hepatoprotective activity, serum enzymes (AST, ALT and alkaline phosphatase) indicative of liver damage were measured and histological and morphological analyses of liver tissue were performed. The results obtained showed that asiaticoside exerted hepatoprotective action, since it promoted a reduction in histological lesions and a decrease in serum levels of AST and ALT. From these results, we conclude that asiaticoside, in the dose most commonly used in herbal medicine, protects the liver against acute hepatitis induced by paracetamol...

O asiaticosídeo é um triterpenóide presente no extrato da Centella asiatica, sendo responsável pela atividade terapêutica desta planta em doenças hepáticas crônicas. O hepatócito é a célula responsável pelas funções endócrinas e exócrinas do fígado, além de converter substâncias nocivas em materiais não tóxicos excretados pela bile. Por esse motivo, o fígado é sensível à ação de alguns fármacos, como, por exemplo, o paracetamol. Assim, utilizando o paracetamol como modelo experimental de lesão hepática, o objetivo deste estudo foi avaliar a ação hepatoprotetora do asiaticosídeo, na dose estabelecida como terapêutica, em ratos Wistar. Dois grupos compostos por vinte animais cada, tratados com asiaticosídeo (1mg/kg/dia v.o.) por oito dias e não tratados foram submetidos à intoxicação com elevada dose de paracetamol (3 g/kg v.o) no oitavo dia. Em seguida, os animais foram eutanasiados após 24 h ou 72 h para coleta de amostras de sangue e fragmentos de fígado. Para avaliação da atividade hepatoprotetora, foi realizada a dosagem sérica de enzimas indicativas de lesão hepática (AST, ALT e Fosfatase Alcalina) e a análise histológica e morfométrica do tecido hepático. Os resultados obtidos permitiram evidenciar que na dose utilizada, o asiaticosídeo apresenta atividade hepatoprotetora, uma vez que o grupo submetido ao tratamento prévio apresentou menos lesões histológicas e menores níveis séricos de AST e ALT quando comparado ao grupo controle. Estes resultados permitem concluir que o asiaticosídeo, na dose mais usualmente empregada na fitoterapia, apresentou atividade hepatoprotetora na hepatite aguda causada por elevada dose de paracetamol...

Study of constructed wetlands effluent disinfected with ozone.

The purpose of this research was to study the disinfection of sanitary effluent from constructed wetlands, evaluating the oxidation of organic matter, the formation of formaldehyde, as well as the efficiency of total coliforms and Escherichia coli inactivation. A constant flow of ozone was applied to the batch system in 5 and 10 mg.O3 L(-1) doses with contact times of 5 and 10 min. This study revealed that the average values of formaldehyde formation ranged between 259.00 and 379.00 µg L(-1), which means that the values are within World Health Organization recommended values. The total coliforms and E. coli showed complete inactivation in almost all tests. The dose of ozone 5 mg.O3 L(-1) and contact time of 5 min were sufficient for a significant reduction of the concentration levels of pathogens in constructed wetlands effluent with similar characteristics, thus allowing for its agricultural reuse.

Integrated assessment for establishing an oil environmental vulnerability map: case study for the Santos Basin region, Brazil.

The growth of maritime transport and oil exploitation activities may increase the risk of oil spills. Thus, plans and actions to prevent or mitigate impacts are needed to minimize the effects caused by oil. However, tools used worldwide to support contingency plans have not been integrated, thus leading to failure in establishing priority areas. This investigation aimed to develop indices of environmental vulnerability to oil (IEVO), by combining information about environmental sensibility to oil and results of numerical modeling of spilled oil. To achieve that, a case study concerning to oil spills scenarios in a subtropical coastal area was designed, and IEVOs were calculated and presented in maps, in order to make the information about the areas' vulnerability more easily visualized. For summer, the extension of coastline potentially affected by oil was approximately 150 km, and most of the coastline presented medium to high vulnerability. For winter, 230 km coastline would be affected, from which 75% were classified as medium to high vulnerability. Thus, IEVO maps allowed a rapid and clearer interpretation of the vulnerability of the mapped region, facilitating the planning process and the actions in response to an oil spill.

The I-CreI meganuclease and its engineered derivatives: applications from cell modification to gene therapy.

Meganucleases (MNs) are highly specific enzymes that can induce homologous recombination in different types of cells, including mammalian cells. Consequently, these enzymes are used as scaffolds for the development of custom gene-targeting tools for gene therapy or cell-line development. Over the past 15 years, the high resolution X-ray structures of several MNs from the LAGLIDADG family have improved our understanding of their protein-DNA interaction and mechanism of DNA cleavage. By developing and utilizing high-throughput screening methods to test a large number of variant-target combinations, we have been able to re-engineer scores of I-CreI derivatives into custom enzymes that target a specific DNA sequence of interest. Such customized MNs, along with wild-type ones, have allowed for exploring a large range of biotechnological applications, including protein-expression cell-line development, genetically modified plants and animals and therapeutic applications such as targeted gene therapy as well as a novel class of antivirals.

Anaerobic effluent disinfection using ozone: byproducts formation.

This research was aimed at studying oxidation processes, coliform inactivation effectiveness and disinfection byproducts (DBPs) associated with the disinfection of anaerobic sanitary wastewater effluent with ozone applied at doses of 5.0, 8.0 and 10.0mg O(3)L(-1) for contact times of 5, 10 and 15 min. The wastewater used in this research was generated by the wastewater treatment plant (WWTP), University of São Paulo - Brazil. The total coliform inactivation range was 2.00-4.06 log(10), and the inactivation range for Escherichia coli was 2.41-4.65 log(10). Mean chemical oxygen demand (COD) reductions were 37.6%, 48.8% and 42.4% for doses of 5.0, 8.0 and 10.0mg O(3)L(-1), respectively. Aldehyde formation varied with dosage only when the ozone dose was increased from 5.0 to 8.0mg O(3)L(-1) for acetaldehyde and from 5.0 to 8.0 and from 8.0 to 10.0mg O(3)L(-1) for glyoxal.

Lung tissue mechanics and extracellular matrix remodeling in acute lung injury.

UNLABELLED: This study was undertaken to test whether there is structural remodeling of lung parenchyma that could lead to tissue mechanical changes at an early phase of varying degrees of acute lung injury (ALI). Tissue resistance (R), dynamic elastance (E), and hysteresivity (eta) were analyzed during sinusoidal oscillations of rat lung parenchymal strips 24 h after intraperitoneal injection of saline (C) or paraquat (P [10, 15, 25, and 30 mg/kg]). These strips were also stained in order to quantify the amount of collagen and of three types of elastic fibers (elaunin, oxytalan, and fully developed elastic fibers) in the alveolar septa. E augmented progressively from C to P25, but the data from the P25 and P30 groups were not different (p < 0.0001). R and eta increased from C to P10 and from P15 to P25 (p < 0.001). Collagen fiber content increased exponentially with the severity of the injury. Elaunin and fully developed elastic fibers remained unchanged in the five groups, while oxytalan fibers increased only in the P25 and P30 groups. In conclusion, the pronounced mechanical changes at the tissue level and fibroelastogenesis happened at an early phase of the disease and even in mildly abnormal lung parenchyma. KEYWORDS: elastance; collagen fibers; elastin; paraquat

Neutralization of the pharmacological effects of bothropstoxin-I from Bothrops jararacussu (jararacuçu) venom by crotoxin antiserum and heparin.

Bothropstoxin-I (BthTX-I), the principal myotoxin of Bothrops jararacussu venom, is devoid of phospholipase A(2) (PLA(2)) activity but capable of blocking neuromuscular transmission in mouse nerve-muscle preparations. In this study, the ability of crotoxin antiserum and heparin in preventing the neurotoxic and myotoxic effects of BthTX-I was investigated. Phrenic nerve-diaphragm preparations (PND) stimulated indirectly with supramaximal stimuli (0.2 ms, 0.1 Hz) were incubated with BthTX-I (20 microg/ml) alone or with BthTX-I preincubated with antiserum or heparin for 30 min at 37 degrees C prior to testing. Control preparations were incubated with Tyrode solution, antiserum or heparin alone. BthTX-I (20 microg/ml) produced 50% neuromuscular blockade in the PND preparations in 31+/-4min, with complete blockade occurring in 120 min. The antiserum and heparin significantly prevented the neuromuscular blockade caused by BthTX-I (84 +/- 4% and 100% protection, respectively). Light microscopy examination of the muscles at the end of the 120 min incubation showed that BthTX-I damaged 48 +/- 6% of the fibers. Preincubating the toxin with antivenom significantly reduced the extent of this damage (only 15 +/- 4% of fibers affected, corresponding to 69% protection, P<0.01) whereas heparin offered no protection (34 +/- 7% of fibers affected, not significantly different from that seen with toxin alone). These results show that the antivenom was more effective in neutralizing the myotoxic effects of BthTX-I than was heparin.

Lung tissue mechanics and extracellular matrix composition in a murine model of silicosis.

The dynamic mechanical properties of lung tissue and its contents of collagen and elastic fibers were studied in strips prepared from mice instilled intratracheally with saline (C) or silica [15 (S15) and 30 days (S30) after instillation]. Resistance, elastance, and hysteresivity were studied during oscillations at different frequencies on S15 and S30. Elastance increased from C to silica groups but was similar between S15 and S30. Resistance was augmented from C to S15 and S30 and was greater in S30 than in S15 at higher frequencies. Hysteresivity was higher in S30 than in C and S15. Silica groups presented a greater amount of collagen than did C. Elastic fiber content increased progressively along time. This increment was related to the higher amount of oxytalan fibers at 15 and 30 days, whereas elaunin and fully developed elastic fibers were augmented only at 30 days. Silicosis led not only to pulmonary fibrosis but also to fibroelastosis, thus assigning a major role to the elastic system in the silicotic lung.

Crystal structure of the thermostable archaeal intron-encoded endonuclease I-DmoI.

I-DmoI is a 22 kDa endonuclease encoded by an intron in the 23 S rRNA gene of the hyperthermophilic archaeon Desulfurococcus mobilis. The structure of I-DmoI has been determined to 2.2 A resolution using multi-wavelength anomalous diffraction techniques. I-DmoI, a protein of the LAGLIDADG motif family, represents the first structure of a freestanding endonuclease with two LAGLIDADG motifs, and the first of a thermostable homing endonuclease. I-DmoI consists of two similar alpha/beta domains (alphabetabetaalphabetabetaalpha) related by pseudo 2-fold symmetry. The LAGLIDADG motifs are located at the carboxy-terminal end of the first alpha-helix of each domain. These helices form a two-helix bundle at the interface between the domains and are perpendicular to a saddle-shaped DNA binding surface, formed by two four-stranded antiparallel beta-sheets. Despite substantially different sequences, the overall fold of I-DmoI is similar to that of two other LAGLIDADG proteins for which the structures are known, I-CreI and the endonuclease domain of PI-SceI. The three structures differ most in the loops connecting the beta-strands, relating to the respective DNA target site sizes and geometries. In addition, the absence of conserved residues surrounding the active site, other than those within the LAGLIDADG motif, is of mechanistic importance. Finally, the carboxy-terminal domain of I-DmoI is smaller and has a more irregular fold than the amino-terminal domain, which is more similar to I-CreI, a symmetric homodimeric endonuclease. This is reversed compared to PI-SceI, where the amino-terminal domain is more similar to carboxy-terminal domain of I-DmoI and to I-CreI, with interesting evolutionary implications.

Crystallization and preliminary crystallographic analysis of the archaeal intron-encoded endonuclease I-DmoI.

Two forms of the archaeal intron-encoded site-specific endonuclease I-DmoI, namely I-DmoIc and I-DmoIl, have been purified and crystallized. Crystals of I-DmoIc are rod-shaped and diffract to 3.0 A resolution, but further analysis was hampered by twinning. Crystals of I-DmoIl, which is a six-amino-acid C-terminal truncation of I-DmoIc, are plate shaped and belong to space group C2 with cell parameters a = 93.72, b = 37.03, c = 55.56 A, beta = 113.4 degrees, with one molecule per asymmetric unit (Vm = 2.01 A3 Da-1). The crystals diffract to at least 2.3 A resolution. A complete native data set has been measured and structure determination is on-going.