Mechanisms of programmed cell death associated to severe dengue in human renal lesions.

Dengue virus (DENV) is a global health problem. Severe dengue can manifest with hemorrhage and signs of organ dysfunction, including the kidneys. The innate immune system is an important barrier against arbovirus infection and, specifically in dengue, the cytokines IL1ß and IL18 and caspase-1 activation make up a set of host immune strategies. Cell death mechanisms include pyroptosis, necroptosis and autophagy, each with peculiar markers: gasdermin, RIPK3/MLKL, LC3, respectively. In DENV infection, necrosis and apoptosis are involved and, when infecting monocytes and macrophages in vitro, DENV is capable of inducing pyroptosis. Our objective was to explore the presence of markers of necroptosis, pyroptosis and autophagy in renal lesions caused by DENV. MATERIAL AND METHODS: twenty specimens of lesions from patients who died due to DENV infection, from the pathology department of Hospital Guilherme Álvaro, Santos, SP, were subjected to histological and immunohistochemical studies. Histological sections were stained with hematoxylin-eosin to evaluate tissue changes or collected for research with antibodies: anti-DENV (Instituto Evandro Chagas-PA), RIPK3 (NBP2-45592), MLKL (ab184718), gasdermin D (#36425), LC3 (14600-AP), caspase 1 (#98033), IL1ß (AF201-NA) and IL18 (SC6178). Semi-quantitative analysis was performed on 20 glomeruli and evaluation on tubules and mononuclear cells. This study was approved by the ethics committee of the USP Faculty of Medicine. RESULTS: histological analysis demonstrated glomerular congestion, glomerulitis (medium to severe), acute kidney injury and hyalinization of the glomeruli. Viral antigens were visualized on mononuclear cells. LC3 (autophagy) expression ranged from moderate to intense (++/+++) in glomeruli, tubules and mononuclear cells. The expression of gasdermin (pyroptosis) was mild (+) in most cases in the glomeruli and moderate (++) in the tubules. RIPK3 and MLKL (necroptosis) mild in tubules and mononuclear cells (+). The expression of the cytokines IL1ß and IL18 and caspase 1 was moderate (++). Statistical analysis showed greater expression of LC3 over the others. CONCLUSIONS: Our results contribute to the understanding of the pathogenesis of renal involvement in severe dengue, considering the likely anti-viral mechanism of autophagy. To a lesser extent, pyroptosis is also present, corroborating previous data.

Assessing Spns2-dependent S1P Transport as a Prospective Therapeutic Target.

S1P (sphingosine 1-phosphate) receptor modulator (SRM) drugs interfere with lymphocyte trafficking by downregulating lymphocyte S1P receptors. While the immunosuppressive activity of SRM drugs has proved useful in treating autoimmune diseases such as multiple sclerosis, that drug class is beset by on-target liabilities such as initial dose bradycardia. The S1P that binds to cell surface lymphocyte S1P receptors is provided by S1P transporters. Mice born deficient in one of these, spinster homolog 2 (Spns2), are lymphocytopenic and have low lymph S1P concentrations. Such observations suggest that inhibition of Spns2-mediated S1P transport might provide another therapeutically beneficial method to modulate immune cell positioning. We report here results using a novel S1P transport blocker (STB), SLF80821178, to investigate the consequences of S1P transport inhibition in rodents. We found that SLF80821178 is efficacious in a multiple sclerosis model but - unlike the SRM fingolimod - neither decreases heart rate nor compromises lung endothelial barrier function. Notably, although Spns2 null mice have a sensorineural hearing defect, mice treated chronically with SLF80821178 have normal hearing acuity. STBs such as SLF80821178 evoke a dose-dependent decrease in peripheral blood lymphocyte counts, which affords a reliable pharmacodynamic marker of target engagement. However, the maximal reduction in circulating lymphocyte counts in response to SLF80821178 is substantially less than the response to SRMs such as fingolimod (50% vs. 90%) due to a lesser effect on T lymphocyte sub-populations by SLF80821178. Finally, in contrast to results obtained with Spns2 deficient mice, lymph S1P concentrations were not significantly changed in response to administration of STBs at doses that evoke maximal lymphopenia, which indicates that current understanding of the mechanism of action of S1P transport inhibitors is incomplete.

Moderate physical exercise and ATP modulate the P2X7 receptor and improve cisplatin-induced gastric emptying delay in rats.

Patients undergoing chemotherapy with cisplatin commonly present gastrointestinal effects such as constipation and gastric emptying (GE) delay. Both the purinergic system and physical exercise modulate the gastrointestinal (GI) tract. In the current study, we investigated the role of ATP, physical exercise, and P2X7 receptor blocking on GE delay induced by cisplatin in rats. Male rats were divided into the following groups: control (C), cisplatin (Cis), exercise (Ex), Brilliant Blue G (BBG), ATP, Cis+Ex, Cis+ATP, Cis+BBG, Cis+Ex+BBG, Cis+Ex+BBG+ATP, and Cis+ATP+BBG. GE delay was induced by treatment with 1 mg/kg cisplatin (1 time/week for 5 weeks, ip). The moderate physical exercise was swimming (1 h/day, 5 days/week for 5 weeks). At the end of the treatment or exercise and 30 min before the GE assessment, some groups received BBG (50 mg/kg, sc) or ATP (2 mg/kg, sc). Then, GE was assessed after a 10-min postprandial period. Chronic use of Cis decreased GE delay (P<0.05) compared to the control group. Both exercise and ATP prevented (P<0.05) GE delay compared to Cis. The pretreatment with BBG significantly inhibited (P<0.05) the effect of exercise and ATP. On the other hand, the association between exercise and ATP reversed (P<0.05) the effect of the BBG and prevented GE delay. Therefore, we suggest that both exercise and treatment with ATP activate P2X7 receptors and prevent GE delay induced by cisplatin in rats.

Moderate physical exercise and ATP modulate the P2X7 receptor and improve cisplatin-induced gastric emptying delay in rats

Patients undergoing chemotherapy with cisplatin commonly present gastrointestinal effects such as constipation and gastric emptying (GE) delay. Both the purinergic system and physical exercise modulate the gastrointestinal (GI) tract. In the current study, we investigated the role of ATP, physical exercise, and P2X7 receptor blocking on GE delay induced by cisplatin in rats. Male rats were divided into the following groups: control (C), cisplatin (Cis), exercise (Ex), Brilliant Blue G (BBG), ATP, Cis+Ex, Cis+ATP, Cis+BBG, Cis+Ex+BBG, Cis+Ex+BBG+ATP, and Cis+ATP+BBG. GE delay was induced by treatment with 1 mg/kg cisplatin (1 time/week for 5 weeks, ip). The moderate physical exercise was swimming (1 h/day, 5 days/week for 5 weeks). At the end of the treatment or exercise and 30 min before the GE assessment, some groups received BBG (50 mg/kg, sc) or ATP (2 mg/kg, sc). Then, GE was assessed after a 10-min postprandial period. Chronic use of Cis decreased GE delay (P<0.05) compared to the control group. Both exercise and ATP prevented (P<0.05) GE delay compared to Cis. The pretreatment with BBG significantly inhibited (P<0.05) the effect of exercise and ATP. On the other hand, the association between exercise and ATP reversed (P<0.05) the effect of the BBG and prevented GE delay. Therefore, we suggest that both exercise and treatment with ATP activate P2X7 receptors and prevent GE delay induced by cisplatin in rats.

Comet assay to evaluate chromosomal changes in chickens (Gallus gallus domesticus) contaminated by lead in a city in Bahia.

Chicken (Gallus gallus domesticus) is one of the primary sources of animal protein for the Brazilian population. Thus, the safety of this food is highly relevant. This study was based on the evidence of severe contamination of these animals by metals such as lead in Santo Amaro, Bahia. This exploratory study aimed to evaluate associations between lead levels in blood of chicken exposed to a contaminated area with the occurrence of chromosomal alterations, evidencing genotoxic effects. Serum lead analysis was performed by GF-AAS after dilution with a matrix modifier solution (Triton X-100 0.2% v/v and HNO3 0.1% v/v), while chromosomal damage was evaluated using the comet assay. The results showed genotoxic effects (positive comet assay) only for the specimen sample with higher serum lead concentrations (33.9 µg dL-1), suggesting the occurrence of toxic effects at this level of exposure. This work evaluated a relationship between the reduction of serum lead levels in chicken and increased distance from the primary polluting source - a lead processing plant (COBRAC). It also showed that lead is bioavailable in this territory, contaminating chicken and causing genotoxic effects in these animals, further expanding the concern with the local biota and the health of the residents of Santo Amaro.

Quantitative analysis and phase assemblage of basic oxygen furnace slag hydration.

Basic oxygen furnace (BOF) slag from steelmaking could be applied as a binder in building materials, reducing the CO2 footprint and solid waste, which is relevant for industrial waste management and circular economy. However, its use is mostly restricted because its hydraulic activity is poorly understood. The BOF slag was hydrated in this study, and its reaction products were systematically characterized using XRD, QXRD, and SEM/EDX-based phase mapping. Internal consistency checks of the data were performed between the analytical techniques. The results revealed that the composition of the amorphous hydration products could be identified and quantified, and the main hydration products were hydrogarnets and C-S-H gel. An extended milling process significantly improved the reactivity, and all the major slag phases, including wüstite, participated in the reaction. Brownmillerite formed hydrogarnets during the first 7 days of hydration. The new hydration products contributed to the immobilization of vanadium and chromium. Particle size played an important role in the amount of C2S reacting, the composition of the hydrogarnets and C-S-H gel, their proportions, and the immobilization capacity. Based on the findings, an overall hydration reaction was developed.

Postmenopausal women's cognitive function and performance of virtual reality tasks.

OBJECTIVE: This study aimed to assess whether prior knowledge of computer use determines performance of virtual reality tasks by postmenopausal women and whether menopausal symptoms, sociodemographic factors, lifestyle and cognition modify or interfere with their performance. METHOD: This cross-sectional study included 152 postmenopausal women divided into two groups: computer users and non-users. Age, ethnicity, time of menopause, menopausal symptoms, female health status, level of physical activity and cognitive function were considered. The participants played a virtual reality game and were assessed for hits, errors, omissions and game time. The Mann-Whitney, chi-square and Fisher exact tests and multivariate linear regression analysis were used. RESULTS: Postmenopausal computer users play virtual reality games (p = 0.005) better than postmenopausal non-users of computers. Vasomotor symptoms were high in women who used computers compared to those who did not (p = 0.006). Multivariate linear regression analysis found that the best-fitting predictors for the number of hits - that is, age (p = 0.039), Mini-Mental State Examination score (p = 0.006) and the headache symptom (p = 0.021) - influence the performance of virtual reality tasks. CONCLUSION: Computer users performed virtual reality tasks better than non-users. Headache and age but not vasomotor symptoms negatively affected the postmenopausal women's performance.

Different litter compositions influence broiler chicken locomotion.

Background: Bedding material must absorb moisture, reduce impacts, and allow chicken to express their natural behavior reducing the occurrence of injuries in the footpad and joints, and improving carcass quality and performance. Aims: This study evaluated different bedding materials with different levels of inclusion of dried grass (Zoysia japonica) on the development of lesions in the locomotor system of broiler chickens. Methods: One thousand eight-day-old male chicks of the Cobb 500® were distributed in a completely randomized design with a 3 × 2 factorial scheme: three levels of grass inclusion and two bedding materials (100% wood shavings; 100% rice husks; 25% grass and 75% wood shavings; 25% grass and 75% rice husks; 50% grass and 50% wood shavings; 50% grass and 50% rice husks). The birds at 21, 28, 35, and 42 days of age were evaluated for pododermatitis. Ten birds from each repetition were identified and evaluated weekly from 21 to 42 days for Gait score, latency to lie and leg angle (valgus and varus). At 43 days of age, 60 birds of each treatment were slaughtered and evaluated for femoral degeneration and tibial dyschondroplasia. Results: Different litter compositions did not affect the incidence of lameness, tibial dyschondroplasia and spondylolisthesis. Pododermatitis scores increased after 28 days when including 50% of grass. Conclusion: Age contributes more to the development of locomotion injuries than does bedding material. It is recommended to use inclusions of dried grass only as bedding for young broilers.

Proteomic analysis reveals rattlesnake venom modulation of proteins associated with cardiac tissue damage in mouse hearts.

Snake envenomation is a common but neglected disease that affects millions of people around the world annually. Among venomous snake species in Brazil, the tropical rattlesnake (Crotalus durissus terrificus) accounts for the highest number of fatal envenomations and is responsible for the second highest number of bites. Snake venoms are complex secretions which, upon injection, trigger diverse physiological effects that can cause significant injury or death. The components of C. d. terrificus venom exhibit neurotoxic, myotoxic, hemotoxic, nephrotoxic, and cardiotoxic properties which present clinically as alteration of central nervous system function, motor paralysis, seizures, eyelid ptosis, ophthalmoplesia, blurred vision, coagulation disorders, rhabdomyolysis, myoglobinuria, and cardiorespiratory arrest. In this study, we focused on proteomic characterization of the cardiotoxic effects of C. d. terrificus venom in mouse models. We injected venom at half the lethal dose (LD50) into the gastrocnemius muscle. Mouse hearts were removed at set time points after venom injection (1 h, 6 h, 12 h, or 24 h) and subjected to trypsin digestion prior to high-resolution mass spectrometry. We analyzed the proteomic profiles of >1300 proteins and observed that several proteins showed noteworthy changes in their quantitative profiles, likely reflecting the toxic activity of venom components. Among the affected proteins were several associated with cellular deregulation and tissue damage. Changes in heart protein abundance offer insights into how they may work synergistically upon envenomation. SIGNIFICANCE: Venom of the tropical rattlesnake (Crotalus durissus terririficus) is known to be neurotoxic, myotoxic, nephrotoxic and cardiotoxic. Although there are several studies describing the biochemical effects of this venom, no work has yet described its proteomic effects in the cardiac tissue of mice. In this work, we describe the changes in several mouse cardiac proteins upon venom treatment. Our data shed new light on the clinical outcome of the envenomation by C. d. terrificus, as well as candidate proteins that could be investigated in efforts to improve current treatment approaches or in the development of novel therapeutic interventions in order to reduce mortality and morbidity resulting from envenomation.

Blue and red light photoemitters as approach to inhibit Staphylococcus aureus and Pseudomonas aeruginosa growth / Fotoemissores de luz azul e vermelha como abordagem para inibir o crescimento de Staphylococcus aureus e Pseudomonas aeruginosa

The ability of pathogenic bacteria acquire resistance to the existing antibiotics has long been considered a dangerous health risk threat. Currently, the use of visible light has been considered a new approach to treat bacterial infections as an alternative to antibiotics. Herein, we investigated the antimicrobial effect of two range of visible light, blue and red, on Staphylococcus aureus and Pseudomonas aeruginosa, two pathogenic bacterial commonly found in healthcare settings-acquired infections and responsible for high rate of morbidity and mortality. Bacterial cultures were exposed to blue or red light (470 nm and 660 nm) provided by light-emitting diodes - LED. The fluencies and irradiance used for blue and red light were 284.90 J/cm², 13.19 mW/cm² and 603.44 J/cm², 27.93 mW/cm² respectively. Different experimental approaches were used to determine the optimal conditions of light application. Only exposure to blue light for 6 hours was able to inhibit about 75% in vitro growth of both bacterial species after 24 hours. The surviving exposed bacteria formed colonies significantly smaller than controls, however, these bacteria were able to resume growth after 48 hours. Blue light was able to inhibit bacterial growth upon inoculation in both saline solution and BHI culture medium. We can conclude that blue light, but not red light, is capable of temporarily retarding the growth of gram negative and gram positive bacteria.

A capacidade das bactérias patogênicas adquirirem resistência aos antibióticos existentes há muito tempo é considerada uma ameaça perigosa à saúde. Atualmente, o uso da luz visível tem sido considerado uma nova abordagem no tratamento de infecções bacterianas como alternativa aos antibióticos. Neste trabalho, investigamos o efeito antimicrobiano de duas faixas de luz visível, azul e vermelha, em Staphylococcus aureus e Pseudomonas aeruginosa, duas bactérias patogênicas comumente encontradas em infecções adquiridas em instituições de saúde e responsáveis por alta taxa de morbimortalidade. As culturas bacterianas foram expostas à luz azul ou vermelha(470 nm e 660 nm) fornecida por diodos emissores de luz - LED. As fluências e irradiâncias utilizadas para luz azule vermelha foram 284,90 J/cm², 13,19 mW/cm² e 603,44 J/cm², 27,93 mW/cm², respectivamente. Várias abordagens experimentais foram utilizadas para determinar as condições ótimas de aplicação da luz. Apenas a exposição à luz azul por 6 horas foi capaz de inibir cerca de 75% o crescimento in vitro de ambas as espécies bacterianas após24 horas. As bactérias expostas sobreviventes formaram colônias com um tamanho significativamente menor do que os controles, contudo, essas bactérias conseguiram retomar o crescimento normal após 48 horas. A luz azul foi capaz de inibir o crescimento das bactérias após sua inoculação em solução salina ou no meio de cultura rico em nutrientes BHI. Podemos concluir que a luz azul mas não a luz vermelha é capaz de retardar temporariamente o crescimento de bactérias Gram-negativas e Gram-positivas.

Blue and red light photoemitters as approach to inhibit Staphylococcus aureus and Pseudomonas aeruginosa growth

Abstract The ability of pathogenic bacteria acquire resistance to the existing antibiotics has long been considered a dangerous health risk threat. Currently, the use of visible light has been considered a new approach to treat bacterial infections as an alternative to antibiotics. Herein, we investigated the antimicrobial effect of two range of visible light, blue and red, on Staphylococcus aureus and Pseudomonas aeruginosa, two pathogenic bacterial commonly found in healthcare settings-acquired infections and responsible for high rate of morbidity and mortality. Bacterial cultures were exposed to blue or red light (470 nm and 660 nm) provided by light-emitting diodes - LED. The fluencies and irradiance used for blue and red light were 284.90 J/cm2, 13.19 mW/cm2 and 603.44 J/cm2, 27.93 mW/cm2 respectively. Different experimental approaches were used to determine the optimal conditions of light application. Only exposure to blue light for 6 hours was able to inhibit about 75% in vitro growth of both bacterial species after 24 hours. The surviving exposed bacteria formed colonies significantly smaller than controls, however, these bacteria were able to resume growth after 48 hours. Blue light was able to inhibit bacterial growth upon inoculation in both saline solution and BHI culture medium. We can conclude that blue light, but not red light, is capable of temporarily retarding the growth of gram negative and gram positive bacteria.

Resumo A capacidade das bactérias patogênicas adquirirem resistência aos antibióticos existentes há muito tempo é considerada uma ameaça perigosa à saúde. Atualmente, o uso da luz visível tem sido considerado uma nova abordagem no tratamento de infecções bacterianas como alternativa aos antibióticos. Neste trabalho, investigamos o efeito antimicrobiano de duas faixas de luz visível, azul e vermelha, em Staphylococcus aureus e Pseudomonas aeruginosa, duas bactérias patogênicas comumente encontradas em infecções adquiridas em instituições de saúde e responsáveis por alta taxa de morbimortalidade. As culturas bacterianas foram expostas à luz azul ou vermelha (470 nm e 660 nm) fornecida por diodos emissores de luz - LED. As fluências e irradiâncias utilizadas para luz azul e vermelha foram 284,90 J/cm2, 13,19 mW/cm2 e 603,44 J/cm2, 27,93 mW/cm2, respectivamente. Várias abordagens experimentais foram utilizadas para determinar as condições ótimas de aplicação da luz. Apenas a exposição à luz azul por 6 horas foi capaz de inibir cerca de 75% o crescimento in vitro de ambas as espécies bacterianas após 24 horas. As bactérias expostas sobreviventes formaram colônias com um tamanho significativamente menor do que os controles, contudo, essas bactérias conseguiram retomar o crescimento normal após 48 horas. A luz azul foi capaz de inibir o crescimento das bactérias após sua inoculação em solução salina ou no meio de cultura rico em nutrientes BHI. Podemos concluir que a luz azul mas não a luz vermelha é capaz de retardar temporariamente o crescimento de bactérias Gram-negativas e Gram-positivas.

Blue and red light photoemitters as approach to inhibit Staphylococcus aureus and Pseudomonas aeruginosa growth / Fotoemissores de luz azul e vermelha como abordagem para inibir o crescimento de Staphylococcus aureus e Pseudomonas aeruginosa

The ability of pathogenic bacteria acquire resistance to the existing antibiotics has long been considered a dangerous health risk threat. Currently, the use of visible light has been considered a new approach to treat bacterial infections as an alternative to antibiotics. Herein, we investigated the antimicrobial effect of two range of visible light, blue and red, on Staphylococcus aureus and Pseudomonas aeruginosa, two pathogenic bacterial commonly found in healthcare settings-acquired infections and responsible for high rate of morbidity and mortality. Bacterial cultures were exposed to blue or red light (470 nm and 660 nm) provided by light-emitting diodes - LED. The fluencies and irradiance used for blue and red light were 284.90 J/cm2, 13.19 mW/cm2 and 603.44 J/cm2, 27.93 mW/cm2 respectively. Different experimental approaches were used to determine the optimal conditions of light application. Only exposure to blue light for 6 hours was able to inhibit about 75% in vitro growth of both bacterial species after 24 hours. The surviving exposed bacteria formed colonies significantly smaller than controls, however, these bacteria were able to resume growth after 48 hours. Blue light was able to inhibit bacterial growth upon inoculation in both saline solution and BHI culture medium. We can conclude that blue light, but not red light, is capable of temporarily retarding the growth of gram negative and gram positive bacteria.

A capacidade das bactérias patogênicas adquirirem resistência aos antibióticos existentes há muito tempo é considerada uma ameaça perigosa à saúde. Atualmente, o uso da luz visível tem sido considerado uma nova abordagem no tratamento de infecções bacterianas como alternativa aos antibióticos. Neste trabalho, investigamos o efeito antimicrobiano de duas faixas de luz visível, azul e vermelha, em Staphylococcus aureus e Pseudomonas aeruginosa, duas bactérias patogênicas comumente encontradas em infecções adquiridas em instituições de saúde e responsáveis por alta taxa de morbimortalidade. As culturas bacterianas foram expostas à luz azul ou vermelha (470 nm e 660 nm) fornecida por diodos emissores de luz - LED. As fluências e irradiâncias utilizadas para luz azul e vermelha foram 284,90 J/cm2, 13,19 mW/cm2 e 603,44 J/cm2, 27,93 mW/cm2, respectivamente. Várias abordagens experimentais foram utilizadas para determinar as condições ótimas de aplicação da luz. Apenas a exposição à luz azul por 6 horas foi capaz de inibir cerca de 75% o crescimento in vitro de ambas as espécies bacterianas após 24 horas. As bactérias expostas sobreviventes formaram colônias com um tamanho significativamente menor do que os controles, contudo, essas bactérias conseguiram retomar o crescimento normal após 48 horas. A luz azul foi capaz de inibir o crescimento das bactérias após sua inoculação em solução salina ou no meio de cultura rico em nutrientes BHI. Podemos concluir que a luz azul mas não a luz vermelha é capaz de retardar temporariamente o crescimento de bactérias Gram-negativas e Gram-positivas.

Blue and red light photoemitters as approach to inhibit Staphylococcus aureus and Pseudomonas aeruginosa growth.

The ability of pathogenic bacteria acquire resistance to the existing antibiotics has long been considered a dangerous health risk threat. Currently, the use of visible light has been considered a new approach to treat bacterial infections as an alternative to antibiotics. Herein, we investigated the antimicrobial effect of two range of visible light, blue and red, on Staphylococcus aureus and Pseudomonas aeruginosa, two pathogenic bacterial commonly found in healthcare settings-acquired infections and responsible for high rate of morbidity and mortality. Bacterial cultures were exposed to blue or red light (470 nm and 660 nm) provided by light-emitting diodes - LED. The fluencies and irradiance used for blue and red light were 284.90 J/cm2, 13.19 mW/cm2 and 603.44 J/cm2, 27.93 mW/cm2 respectively. Different experimental approaches were used to determine the optimal conditions of light application. Only exposure to blue light for 6 hours was able to inhibit about 75% in vitro growth of both bacterial species after 24 hours. The surviving exposed bacteria formed colonies significantly smaller than controls, however, these bacteria were able to resume growth after 48 hours. Blue light was able to inhibit bacterial growth upon inoculation in both saline solution and BHI culture medium. We can conclude that blue light, but not red light, is capable of temporarily retarding the growth of gram negative and gram positive bacteria.

BPA toxicity during development of zebrafish embryo.

Bisphenol A (BPA) is a monomer used in the production of polycarbonate, a polymer commonly found in plastics, epoxy resins and thermal papers. The presence of BPA in food, water, air and dust has been of great concern in recent years not only due to environmental and ecological issues but also because of its supposed risk to public health related to its mutagenic and carcinogenic potential. In this study we evaluated the toxicity of bisphenol A in zebrafish embryos (Danio rerio) and determined the 50% lethal concentration (LC50) of this chemical. BPA was used at concentrations ranging from 1 µM to 100 µM in E3 medium/0.5% dimethylsulfoxide (DMSO) from previously prepared stock solutions in 100% DMSO. Controls included embryos exposed only to E3 medium or supplemented with 0.5% DMSO. Camptothecin (CPT), a known inhibitor of cell proliferation was used as positive control at a concentration of 0.001 µM in E3 medium/0.5% DMSO. Adults zebrafish were placed for breeding a day before the experimental set up, then, viable embryos were collected and selected for use. Experiments were carried out in triplicates, according to specifications from Organization for Economic Cooperation and Development (OECD). One embryo/well (25 embryos per concentration) was distributed in 96 well microplates in presence or absence of the chemicals. The plates were kept in BOD incubators with a controlled temperature of 28.5 ºC and with photoperiod of 14 h light:10 h dark. After 24h, 48h, 72h and 96h exposure, the exposed embryos were evaluated according to the following parameters: mortality, coagulation, rate of heartbeat, hatching and presence of morphological abnormalities. Photography was obtained by photomicroscopy. Apoptosis was evaluated by DNA ladder assay. DNA was extracted by phenol:chloroform method and analyzed by 2% agarose gel electrophoresis. DNA fragments were visualized after ethidium bromide staining in ultraviolet transilluminator. The LC50 determined for BPA was 70 µM after 24 hours, 72 µM after 48 hours, 47 µM after 72 hours and 31 µM after 96 hours exposure. BPA induced morphological and physiological alterations such as yolk sac and pericardial edema, hatching delay or inhibition, spine deformation, decreasing in heartbeat rate and mortality. In conclusion, this study demonstrated that BPA induced marked malformations in zebrafish embryos at concentrations above 25 µM corroborating the current concerns related to the widespread presence of BPA in the air, food and water used by humans as well as in the bodily fluids and tissues.

Staphylococcus aureus growth delay after exposure to low fluencies of blue light (470 nm).

Antibiotic resistance is one of the greatest challenges to treat bacterial infections worldwide, leading to increase in medical expenses, prolonged hospital stay and increased mortality. The use of blue light has been suggested as an innovative alternative to overcome this problem. In this study we analyzed the antibacterial effect of blue light using low emission parameters on Staphylococcus aureus cultures. In vitro bacterial cultures were used in two experimental approaches. The first approach included single or fractionated blue light application provided by LED emitters (470 nm), with the following fluencies: 16.29, 27.16 and 54.32 J/cm2. For the second approach a power LED (470 nm) was used to deliver 54.32 J/cm2 fractionated in 3 applications. Our results demonstrated that bacterial cultures exposed to fractionated blue light radiation exhibited significantly smaller sizes colonies than the control group after 24 h incubation, however the affected bacteria were able to adapt and continue to proliferate after prolonged incubation time. We could conclude that the hypothetical clinical use of low fluencies of blue light as an antibacterial treatment is risky, since its action is not definitive and proves to be ineffective at least for the strain used in this study.