RA33, an analogue of resveratrol, improves the development of in vitro-fertilized bovine embryos.

Oxidative stress is an undesirable effect of in vitro culture, which requires antioxidant supplementation. This study investigated the analogue of resveratrol (RA33) as an alternative to resveratrol, an antioxidant molecule, for the in vitro culture of in vitro-fertilized bovine embryos. The effect of different concentrations of RA33 on embryo development was evaluated and a comparison between RA33 and resveratrol was performed. The cleavage rate was higher (P < 0.05) with 2.5 µM (69.0 ± 4.4%) than at 0, 0.1 or 0.5 µM RA33 (62.1 ± 2.0%, 60.7 ± 5.9% and 56.7 ± 5.8%, respectively). The blastocyst rates on days 7 and 8 post-fertilization with 2.5 µM RA33 (19.4 ± 3.3% and 24.6 ± 3.3%, respectively) were higher (P < 0.05) than for 0 µM (12.4 ± 2.5% and 15.2±2.5%, respectively). When 2.5 µM RA33 was compared with 0.5 µM resveratrol, similar (P > 0.05) cleavage and blastocyst rates were found between them, but the cleavage rate was higher (P < 0.05) in the control (80.8 ± 3.4%) than for the resveratrol treatment (76.4 ± 3.6%). The numbers of apoptotic cells and the apoptotic index were lower (P < 0.05) with RA33 (6.5 ± 0.6 cells and 6.4 ± 0.7%, respectively) and resveratrol (5 ± 0.8 cells and 5.5 ± 1.0%, respectively) than in the control group (9.8 ± 1.2 cells and 8.9 ± 1.1%, respectively). In conclusion, RA33 can enhance the preimplantation development of in vitro-fertilized bovine embryos and be an alternative to resveratrol in embryo culture medium.

Assessment of screening programs as a strategy for early detection of oral cancer: a systematic review.

The present study is a systematic review of the evaluation of screening programs as a strategy for early detection of oral cancer. The aim of this study was to assess whether screening through visual inspection is able to identify injuries in early stages, to increase survival, and to decrease the incidence and mortality of oral cancer. Studies using visual inspection to screen for oral cancer and potentially malignant lesions in apparently healthy individuals over 18 years without previous diagnosis of the disease were included. The MEDLINE/PubMed, Cochrane databases Library, EMBASE, and LILACS, including manual search and gray literature, were searched through January 2021 with no language or date restrictions. The risk of bias and the methodological quality were evaluated according to the appropriate tool for each study design. The analysis of the results was narrative. Seventeen studies were reviewed that included cohort, accuracy, and randomized clinical trial studies. The tracking type performed was opportunistic and organized in a variety of environments. The age of participants ranged between 18 and 60 years old and, in some programs, only people with risk habits for oral cancer were included. The screeners were healthcare professionals, physicians, and dentists. Two studies reported data on the incidence rate of severe cases and mortality and showed a reduction when patients were at risk for the disease and participated in the program more than once. A limitation of this review was the great variability observed in the estimates of the screening effectiveness among the studies, which made comparisons difficult. If a screening program is continuous and able to ensure the inclusion of high-risk individuals, it can contribute to improvement in survival rates with a change of stage and can have a significant impact on incidence and mortality due to the disease. Registration in the Open Science Framebook (OSF) with the osf.io/zg8nr link.

Fate of Tableted Freeze-Dried siRNA Lipoplexes in Gastrointestinal Environment.

The incorporation of siRNA into nanocarriers is mandatory to facilitate its intracellular delivery, as siRNA itself cannot enter cells. However, the incorporation of these nanocarriers into oral, solid dosage forms and their fate in the gastrointestinal environment is yet to be explored. In the present work, the fate of, (i) naked siRNA, (ii) freshly prepared siRNA lipoplexes, and (iii) tableted siRNA lipoplexes, in simulated gastric and intestinal fluids was studied. The siRNA, either released from or protected within the lipoplexes, was quantified by gel electrophoresis and siRNA efficacy was assessed in cell transfection. The freshly prepared lipoplexes kept their siRNA load and transfection efficiency totally preserved during 1 h of incubation in simulated gastric fluid at 37 °C. However, in simulated intestinal fluid, despite no release of siRNA from lipoplexes after 6 h of incubation, gene silencing efficacy was dramatically decreased even after 1 h of exposure. The lipoplexes obtained from tablets efficiently protected siRNA in simulated gastric fluid, thus preserving the gene silencing efficacy, whereas their incubation in simulated intestinal fluid resulted in a marked siRNA release and decreased gene silencing efficacy. These results provided a detailed explanation for understanding the fate of siRNA in gastrointestinal conditions, when simply loaded in lipoplexes or formulated in the form of tablets.

Neuroprotective Effect of siRNA Entrapped in Hyaluronic Acid-Coated Lipoplexes by Intravitreal Administration.

Since the possibility of silencing specific genes linked to retinal degeneration has become a reality with the use of small interfering RNAs (siRNAs), this technology has been widely studied to promote the treatment of several ocular diseases. Despite recent advances, the clinical success of gene silencing in the retina is significantly reduced by inherent anatomical and physiological ocular barriers, and new strategies are required to achieve intraocular therapeutic effectiveness. In this study, we developed lipoplexes, prepared with sodium alginate as an adjuvant and strategically coated with hyaluronic acid (HA-LIP), and investigated the potential neuroprotective effect of these systems in a retinal light damage model. Successful functionalization of the lipoplexes with hyaluronic acid was indicated in the dynamic light scattering and transmission electron microscopy results. Moreover, these HA-LIP nanoparticles were able to protect and deliver siRNA molecules targeting caspase-3 into the retina. After retinal degeneration induced by high light exposure, in vitro and in vivo quantitative reverse transcription-PCR (RT-qPCR) assays demonstrated significant inhibition of caspase-3 expression by HA-LIP. Furthermore, these systems were shown to be safe, as no evidence of retinal toxicity was observed by electroretinography, clinical evaluation or histology.

Assessment of the safety of intravitreal injection of metoprolol tartrate in rabbits.

PURPOSE: To verify the safety of different doses of intravitreal metoprolol tartrate (MT) after intravitreal injection in rabbit eyes. METHODS: Animals were randomly assigned into 2 groups: group I received 50 µg of MT and group II 100 µg of MT. A volume of 0.05 mL of the drug solution was administered through an intravitreal injection, while the control eyes received an equal volume of saline solution. Safety was assessed by clinical observation, electroretinography (ERG) and histological evaluation. RESULTS: No evidence of clinical toxicity was observed. ERG waveforms from the MT treated eyes were similar to those recorded from the control eyes in dark-adapted state, amplitude and the implicit time are similar between the groups in light-adapted state, and their retinas had no signs of toxicity by histological evaluation 7 days after intravitreal injection. CONCLUSIONS: The intravitreal use of metoprolol at 50 and 100 µg dosages does not cause short-term retinal toxicity in rabbits.

Human Stem Cell-Derived Retinal Pigment Epithelial Cells as a Model for Drug Screening and Pre-Clinical Assays Compared to ARPE-19 Cell Line.

BACKGROUND AND OBJECTIVES: Eye diseases have a high socioeconomic impact on society and may be one of the fields in which most stem cell-related scientific accomplishments have been achieved recently. In this context, human Pluripotent Stem Cell (hPSC) technology arises as an important tool to produce and study human Embryonic Stem cell derived-Retinal Pigmented Epithelial Cells (hES-RPE) for several applications, such as cell therapy, disease modeling, and drug screening. The use of this technology in pre-clinical phases attends to the overall population desire for animal-free product development. Here, we aimed to compare hES-RPE cells with ARPE-19, one of the most commonly used retinal pigmented epithelial immortalized cell lines. METHODS AND RESULTS: Functional, cellular and molecular data obtained suggest that hES-RPE cells more closely resembles native RPEs compared to ARPE-19. Furthermore, hES-RPE revealed an interesting robustness when cultured on human Bruch's membrane explants and after exposure to Cyclosporine (CSA), Sirolimus (SRL), Tacrolimus (TAC), Leflunomide (LEF) and Teriflunomide (TER). On these conditions, hES-RPE cells were able to survive at higher drug concentrations, while ARPE-19 cell line was more susceptible to cell death. CONCLUSIONS: Therefore, hES-RPEs seem to have the ability to incur a broader range of RPE functions than ARPE-19 and should be more thoroughly explored for drug screening.

Compression of Vectors for Small Interfering RNAs Delivery: Toward Oral Administration of siRNA Lipoplexes in Tablet Forms.

Currently, most nonviral nucleic acid vectors are in the form of colloidal suspensions administered primarily parenterally. This type of formulation and the mode of administration impose strong constraints such as the size of the administered vectors or the production of sterile preparations. The tablet form provides access to easy oral administration, well accepted by patients; As regards nucleic acid vectors, a dry form represents an advance in terms of stability. Using an optimized lipid-based small interfering RNA-delivery system, we studied the tabletability of a liquid suspension of these vectors. We optimized the conditions of freeze-drying by choosing excipients and process, allowing for the conservation of both the gene-silencing efficacy of the formulated siRNAs and the supramolecular structure of the lipid particulate system. Gene-silencing efficacy was assayed on luciferase-expressing cells and the structure of the siRNA vector in freeze-dried and tablet forms was examined using small-angle X-ray scattering (SAXS) synchrotron radiation. The freeze-dried powders were then mixed with excipients necessary for the good progress of the compression by allowing for a regular supply of the matrix and the reduction of friction. The compression was carried out using a rotary press simulator that allows for complete monitoring of the compression conditions. After compression, formulated siRNAs retained more than 60% of their gene-silencing efficacy. Within the tablets, a specific SAXS signal was detectable and the lamellar and cubic phases of the initial liquid suspension were restored after resuspension of siRNA vectors by disintegration of the tablets. These results show that the bilayer lipid structures of the particles were preserved despite the mechanical constraints imposed by the compression. If such a result could be expected after the freeze-drying step, it was never shown, to our knowledge, that siRNA-delivery systems could retain their efficacy and structure after mechanical stress such as compression. This opens promising perspectives to oral administration of siRNA as an alternative to parenteral administration.

Considerations for cholinesterase biomonitoring in flower and ornamental plant greenhouse workers.

Flower cultivation in greenhouses enhances productivity and ensures high-quality product supply throughout the year. However, under such protected environments, climate conditions are designed to favor flower growth and can intensify occupational exposure to pesticides. In this study, flower greenhouse workers were tested for cholinesterase inhibition, a valuable indicator of occupational exposure to organophosphate and carbamate pesticides. The quantified inhibition was also associated with working practices using these outcomes to explore ways in which provisions of regulatory Occupational Health Control Programs can be met considering the peculiarities of this working environment. Despite their preliminary nature, and considering that workers had not been previously tested, the obtained results suggested that the designed pre- and post-screening process, enabled biomonitoring of the exposed population. The pre-exposure sample was collected after 11 days from pesticide spraying and the post-exposure sample was collected no later than 10 days after pesticide spraying. AChE activity was significantly depressed in flower greenhouse workers, comparing pre- and post-exposure intervals. Nursery workers, often overlooked, presented greater inhibition of AChE activity. Such finding is consistent with effects believed to be associated with the described job and the way activities are conducted, once nursery workers are regularly exposed to low levels of pesticides owing to their continuous contact with sprayed crops. As predicted by Occupational Safety and Health Standards, establishing biomonitoring programs is achievable, despite the intensive use of pesticides. Quantification is important to target the exposed population but is not sufficient to assure good working conditions and avoid exposure. Administrative and working practices, such as job misclassification, non-compliance with national regulations for occupational health control, pesticide handling, lack of REI protocols, and unsuitable delivery of PPE, must be addressed. Anything less will contribute to worsening an already bleak picture in terms of occupational safety and health in flower and ornamental plant greenhouses.

Endo-ß-1,3-glucanase (GH16 Family) from Trichoderma harzianum Participates in Cell Wall Biogenesis but Is Not Essential for Antagonism Against Plant Pathogens.

Trichoderma species are known for their ability to produce lytic enzymes, such as exoglucanases, endoglucanases, chitinases, and proteases, which play important roles in cell wall degradation of phytopathogens. ß-glucanases play crucial roles in the morphogenetic-morphological process during the development and differentiation processes in Trichoderma species, which have ß-glucans as the primary components of their cell walls. Despite the importance of glucanases in the mycoparasitism of Trichoderma spp., only a few functional analysis studies have been conducted on glucanases. In the present study, we used a functional genomics approach to investigate the functional role of the gluc31 gene, which encodes an endo-ß-1,3-glucanase belonging to the GH16 family in Trichoderma harzianum ALL42. We demonstrated that the absence of the gluc31 gene did not affect the in vivo mycoparasitism ability of mutant T. harzianum ALL42; however, gluc31 evidently influenced cell wall organization. Polymer measurements and fluorescence microscopy analyses indicated that the lack of the gluc31 gene induced a compensatory response by increasing the production of chitin and glucan polymers on the cell walls of the mutant hyphae. The mutant strain became more resistant to the fungicide benomyl compared to the parental strain. Furthermore, qRT-PCR analysis showed that the absence of gluc31 in T. harzianum resulted in the differential expression of other glycosyl hydrolases belonging to the GH16 family, because of functional redundancy among the glucanases.

Vivências de mulheres enfrentando a histerectomia: estudo fenomenológico / Living of women facing hysterectomy: phenomenological study / Vivencias de mujeres enfrentando la histerectomia: estudio fenomenológico

Objetivo compreender os significados e desvelar os sentidos do ser mulher que vivencia o pré-operatório de histerectomia. Método: estudo de natureza qualitativa, pautado na fenomenologia heideggeriana. Realizado na Clínica Cirúrgica de um Hospital de Ensino da Zona da Mata Mineira. Foi participante quatorze mulheres em pré-operatório que em entrevista fenomenológica realizaram seus depoimentos nos meses de agosto a outubro de 2017. Resultado: da análise compreensiva emergiu as Unidades de Significação: aceitar fazer a cirurgia e Apoiar-se em Deus e sentir-se bem cuidada no hospital. A compreensão interpretativa desvelou que no cotidiano a mulher se mostra inautêntica, vivenciando a impropriedade e o falatório. Considerações Finais: se faz necessário que a enfermagem acolha o ser-mulher e compreenda sua dimensão existencial, seu estar-no- mundo. É preciso atentar não somente ao seu corpo biológico, mas que pause a mecanicidade de suas atividades para que possa oferecer o cuidado humanístico mediante sua singularidade, subjetividade e demandas.(AU)

Aimed at understanding the meanings and unveil the senses of being a woman who experiences the preoperative hysterectomy. Method: study of a qualitative nature, based on Heideggerian phenomenology. Performed at the Surgical Clinic of a Teaching Hospital of the Mata Mineira Zone. Participants were fourteen preoperative women who in a phenomenological interview conducted their testimonies in the months of August to October 2017. Results: from the comprehensive analysis emerged the Units of Significance: to accept to do the surgery and to support in God and to feel well taken care of in hospital. The interpretive understanding revealed that in the daily life the woman is inauthentic, experiencing the impropriety and the chatter. Final Considerations: it is necessary for nursing to welcome the being-woman and to understand its existential dimension, its being-in-the-world. It is necessary to pay attention not only to its biological body, which pauses the mechanistic of its activities so that it can offer humanistic care through its singularity, subjectivity and demands.(AU)

Objetivo comprender los significados y desvelar los sentidos del ser mujer que vive el preoperatorio de histerectomía. Método: estudio de naturaleza cualitativa, pautado en la fenomenología heideggeriana. Realizado en la Clínica Quirúrgica de un Hospital de Enseñanza de la Zona de la Mata Minera. Fue participante catorce mujeres en preoperatorio que en entrevista fenomenológica realizaron sus testimonios en los meses de agosto a octubre de 2017. Resultados: del análisis comprensivo emergió las Unidades de Significación: aceptar hacer la cirugía y Apoyarse en Dios y sentirse bien en el hospital. La comprensión interpretativa desveló que en el cotidiano la mujer se muestra inauténtica, vivenciando la impropiedad y el falatorio. Consideraciones finales: se hace necesario que la enfermería acoja al ser-mujer y comprenda su dimensión existencial, su estar en el mundo. Es necesario atentar no sólo a su cuerpo biológico, que pause la mecanicidad de sus actividades para que pueda ofrecer el cuidado humanístico mediante su singularidad, subjetividad y demandas.(AU)

Quality of semen in the reproductive cycle of Cachara (Pseudoplatystoma fasciatum) raised in captivity

In this study, we evaluated the semen quality of Cachara (Pseudoplatystoma fasciatum) raised in captivity during its reproductive period. For the evaluation of qualitative parameters, we analyzed: sperm morphology, sperm motility, and sperm vigor;while for the quantitative parameter, we analyzed sperm concentration. In general, the main significant difference was found in March, with a motility rate of 40% and a duration of 39 seconds. The highest mean concentration (1.2x10⁸) was observed in February, as well as the highest percentage of defects in the intermediate part of the spermatozoa (59%). The morphological analysis of spermatozoa showed an average of 24.2% abnormalities in the head and 22.7% in the intermediate part of spermatozoa of P. fasciatumfor the four months studied. We did not find any evident relationship between the climate changes evaluated and the qualitative and quantitative parameters. The greatest difference between the percentage of normal spermatozoa and the sharp increase of defects in the intermediate piece, observed in February, can be explained by the sudden climate change, with a significant decrease in temperature and relative humidity, and increase in precipitation. These findings demonstrate the importance of maintaining an optimum climate interval during the breeding season for P.fasciatum.

Biochemical and Molecular Study of Trichoderma harzianum Enriched Secretome Protein Profiles Using Lectin Affinity Chromatography.

Protein glycosylation is one of the most studied post-translational modifications and has received considerable attention for its critical role in the cell biology of eukaryotic cells. The genus Trichoderma has been extensively studied in the biocontrol of soil-borne fungal phytopathogens. The aim of this study was to identify the proteins secreted from Trichoderma harzianum after interacting with the cell walls of two phytopathogens, Sclerotinia sclerotiorum and Fusarium oxysporum. This study used N-glycoprotein enrichment with a concanavalin A (Con A) affinity column, staining detection differential SDS-PAGE, sequencing by mass spectrometric, and protein identification by comparison with the NCBI database to detect the protein expression of the two resulting secretome samples. The majority of the proteins found in both enriched secretomes belonged to a specific class of carbohydrate-active enzymes (CAZymes), within which glycosyl hydrolases (GHs), glycosyltransferases (GTs), and auxiliary activities (AAs) were identified. In this study was described two proteins that have not been previously reported in the secretomes of Trichoderma, a glycosyltransferase (six-harpin) and a galactose oxidase, belonging to the class of auxiliary activities (AA), classified as an AA subfamily AA5-2.The expression pattern of gene encoding to 17 identified proteins, evaluated by real-time quantitative PCR (RT-qPCR), showed significant difference of expression of some GHs and proteases, suggesting a specific gene expression regulation by T. harzianum in presence of different cell walls of two phytopathogens.

Toxicity and in vivo release profile of sirolimus from implants into the vitreous of rabbits' eyes.

PURPOSE: To assess the in vivo release profile and the retinal toxicity of a poly (lactic-co-glycolic acid) (PLGA) sustained-release sirolimus (SRL) intravitreal implant in normal rabbit eyes. METHODS: PLGA intravitreal implants containing or not SRL were prepared, and the viability of ARPE-19 and hES-RPE human retinal cell lines was examined after 24 and 72 h of exposure to implants. New Zealand rabbits were randomly divided into two groups that received intravitreal implants containing or not SRL. At each time point (1-8 weeks), four animals from the SRL group were euthanized, the vitreous was collected, and drug concentration was calculated. Clinical evaluation of the eyes was performed weekly for 8 weeks after administration. Electroretinography (ERG) was recorded in other eight animals, four for each group, at baseline and at 24 h, 1, 4, 6, and 8 weeks after the injection. ERG was carried out using scotopic and photopic protocols. The safety of the implants was assessed using statistical analysis of the ERG parameters (a and b waves, a and b implicit time, B/A ratio, oscillatory potential, and Naka-Rushton analysis) comparing the functional integrity of the retina between the PLGA and SRL-PLGA groups. After the last electrophysiological assessment, the rabbits were euthanized and retinal histopathology was realized. RESULTS: After 24 and 72 h of incubation with PLGA or SRL-PLGA implants, ARPE-19 and hES-RPE cells showed viability over 70%. The maximum concentration of SRL (199.8 ng/mL) released from the device occurred within 4 weeks. No toxic effects of the implants or increase in the intraocular pressure was observed through clinical evaluation of the eye. ERG responses showed no significant difference between the eyes that received PLGA or SRL-PLGA implants at baseline and throughout the 8 weeks of follow-up. No remarkable difference in retinal histopathology was detected in rabbit eyes treated with PLGA or SRL-PLGA implants. CONCLUSIONS: Intravitreal PLGA or SRL-PLGA implants caused no significant reduction in cell viability and showed no evident toxic effect on the function or structure of the retina of the animals. SRL was released from PLGA implant after application in the vitreous of rabbits during 8 weeks.

Predation and reproductive performance in two pelagic typhloplanid turbellarians.

We investigated feeding and reproductive performance of coexisting pelagic turbellarians from experiments on predation rates of Mesostoma ehrenbergii and M. craci as a function of food (Daphnia similis, three levels) and temperature (4 levels) during 10 days. Flatworms were collected from the pelagic of a subtropical lake in Brazil. Growth was more rapid at higher temperatures: more prey were consumed, and more eggs produced. M. craci and particularly M. ehrenbergii fitted a linear mixed-effects model and showed a type II functional response. M. craci was the more stenothermic of the two. Intrageneric predation also occurred: M. ehrenbergii fed on M. craci, but not vice versa. After a first clutch of subitaneous eggs, M. ehrenbergii produced resting eggs only. In M. craci an intermediate type of eggs hatched some time after release, survived passage through the gut of M. ehrenbergii, but did not resist drying. By primarily selecting cladoceran prey, M. ehrenbergii can make coexistence of both flatworms possible. As population density of M. ehrenbergii increases, it turns to producing resting and non-viable subitaneous eggs, thus limiting its population size. In nature, these processes structure the zooplankton community, while avoiding extinction of prey and predator.

Structural characterization of a new steroidal saponin from Agave angustifolia var. Marginata and a preliminary investigation of its in vivo antiulcerogenic activity and in vitro membrane permeability property.

A new furostane steroidal saponin was isolated from the leaves of Agave angustifolia var. marginata. On the basis of chemical conversions and spectroscopic analyses, its structure was established as 3-[O-ß-d-glucopyranosyl-(1→3)-O-ß-d-glucopyranosyl-(1→3)-O]-[O-6-deoxy-α-l-mannopyranosyl-(1→4)-ß-d-xylopyranosyl-(1→2)-O-ß-d-glucopyranosyl-(1→4)-ß-d-galactopyranosyl)oxy]-(3ß,5α,22α,25R)-26-(ß-d-glucopyranosyloxy)-22-methoxy-furostane (1). Results of preliminary biological investigations indicated that compound 1 showed significant protective effects against induced gastric ulcers using in vivo experimental models and demonstrated negligible toxicity on membrane integrity in the in vitro assays.

Effect of induction therapy in kidney transplantation in sensitive patients: analysis of risks and benefits.

INTRODUCTION: Sensitization is associated with worse clinical outcomes after kidney transplantation (KT), including increased incidence of delayed graft function, acute rejection (AR) and graft loss. OBJECTIVES: To evaluate 1-year efficacy and safety outcomes in sensitized KT recipients receiving antithymocyte globulin (ATG) induction and compare them to non-sensitized patients. METHODS: Deceased donors KT recipients transplanted between January 1998 and December 2009 were divided into 5 groups: control group 1 -n = 89, PRA negative, without induction therapy; control group 2 - n = 94, PRA negative, basiliximab induction; control group 3 - n = 81, PRA negative, ATG induction; test group 4 - n = 64, PRA 1-49%, ATG induction; test group 5 -n = 118, PRA ≥ 50%, ATG induction. RESULTS: There was no difference in the incidence of AR among patients sensitized and non-sensitized, except for group 1, with highest incidence of AR (20.2%,p = 0.006 vs. Group 4 and p = 0.001 vs. group 5). Sensitized patients induced with ATG had higher incidence of citomegalovirus infection when compared with group 2 (26.6% and 14.4% vs. 2.1%). There were no differences in graft and patient survivals. In multivariable analysis, PRA > 50% and ATG induction were not associated with graft loss, death or death-censored graft loss. CONCLUSION: Sensitized patients induced with ATG presented similar or lower incidence of AR when compared with non-sensitized patients not induced. Besides, these patients had similar safety profile and graft and patient survivals at 1 year.

Arsenic toxicity to cladocerans isolated and associated with iron: implications for aquatic environments.

Arsenic is an ametal ubiquitous in nature and known by its high toxicity. Many studies have tried to elucidate the arsenic metabolism in the cell and its impact to plants, animals and human health. In aqueous phase, inorganic arsenic is more common and its oxidation state (As III and As V) depends on physical and chemical environmental conditions. The aim of this study was to evaluate toxicity of arsenic to Daphnia similis and Ceriodaphnia silvestrii, isolated and associated with iron. The results showed differences in toxicity of As III and As V to both species. Effective concentration (EC50) mean values were 0.45 mg L-1 (As III) and 0.54 mg L-1 (As V) for D. similis, and 0.44 mg L-1 (As III) and 0.69 mg L-1 (As V) for C. silvestrii. However, As V IC25 mean value was 0.59 mg L-1, indicating that C. silvestrii has mechanisms to reduce arsenic toxicity. On the other hand, when associated with iron at 0.02 and 2.00 mg L-1, EC50 values decreased for D. similis (0.34 and 0.38 mg L-1) as well as C. silvestrii (0.37 and 0.37 mg L-1), showing synergistic effect of these substances.

Efeito da terapia de indução em pacientes sensibilizados: análise dos riscos e benefícios / Effect of induction therapy in kidney transplantation in sensitive patients: analysis of risks and benefits

Resumo Introdução: A sensibilização está associada a piores desfechos clínicos após o transplante renal (TxR), incluindo maior incidência de função tardia, rejeição aguda e perda do enxerto. Objetivos: Avaliar os desfechos de eficácia e segurança de 1 ano de receptores de TxR com doador falecido sensibilizados induzidos com globulina antitimócito (ATG) e compará-las aos de pacientes não sensibilizados. Métodos: Receptores de TxR com doador falecido entre janeiro de 1998 e dezembro de 2009 foram divididos em 5 grupos: grupo controle 1 - n = 89, PRA negativo, sem indução; grupo controle 2 - n = 94, PRA negativo, indução com basiliximabe; grupo controle 3 - n = 81, PRA negativo, indução com ATG; grupo teste 4 - n = 64, PRA 1-49%, indução com ATG; grupo teste 5 - n = 118, PRA ≥ 50%, indução com ATG. Resultados: Não houve diferença na incidência de rejeição entre pacientes sensibilizados e não sensibilizados, exceto pelo grupo 1, que apresentou a maior incidência de rejeição aguda comprovada por biópsia (20,2%, p = 0,006 vs. grupo 4 ep = 0,001 vs. grupo 5). Os pacientes sensibilizados induzidos com ATG apresentaram maior incidência de infecção por citomegalovírus quando comparados aos pacientes do grupo 2 (26,6% e 14,4% vs. 2,1%). Não houve diferença nas sobrevidas do enxerto e do paciente. Na análise multivariada, PRA > 50% e uso de ATG não foram associados à perda, perda com óbito censorado ou óbito. Conclusão: Os pacientes sensibilizados induzidos com ATG apresentaram incidência de rejeição semelhante ou inferior à de pacientes não sensibilizados não induzidos. Estes pacientes apresentaram sobrevidas do enxerto e do paciente semelhantes em 1 ano e comparável perfil de segurança.

Abstract Introduction: Sensitization is associated with worse clinical outcomes after kidney transplantation (KT), including increased incidence of delayed graft function, acute rejection (AR) and graft loss. Objectives: To evaluate 1-year efficacy and safety outcomes in sensitized KT recipients receiving antithymocyte globulin (ATG) induction and compare them to non-sensitized patients. Methods: Deceased donors KT recipients transplanted between January 1998 and December 2009 were divided into 5 groups: control group 1 -n = 89, PRA negative, without induction therapy; control group 2 - n = 94, PRA negative, basiliximab induction; control group 3 - n = 81, PRA negative, ATG induction; test group 4 - n = 64, PRA 1-49%, ATG induction; test group 5 -n = 118, PRA ≥ 50%, ATG induction. Results: There was no difference in the incidence of AR among patients sensitized and non-sensitized, except for group 1, with highest incidence of AR (20.2%,p = 0.006 vs. Group 4 andp = 0.001 vs. group 5). Sensitized patients induced with ATG had higher incidence of citomegalovirus infection when compared with group 2 (26.6% and 14.4% vs. 2.1%). There were no differences in graft and patient survivals. In multivariable analysis, PRA > 50% and ATG induction were not associated with graft loss, death or death-censored graft loss. Conclusion: Sensitized patients induced with ATG presented similar or lower incidence of AR when compared with non-sensitized patients not induced. Besides, these patients had similar safety profile and graft and patient survivals at 1 year.

Ingasaponin, a complex triterpenoid saponin with immunological adjuvant activity from Inga laurina.

As part of our search of bioactive saponins from Brazilian plants, phytochemical study of the seeds of Inga laurina led to the isolation of a new complex triterpenoid saponin, named ingasaponin. It is the first saponin isolated from a species of Inga genus. It was isolated by using chromatographic methods and its structural elucidation was performed using detailed analyses of (1)H and (13)C NMR spectra including 2D-NMR spectroscopic techniques and chemical conversions. Its structure was established as 21-[[(2E,6S)-6-[[6-deoxy-4-O-[(2E,6S)-6-[(ß-D-glucopyranosyl)oxy]-2,6-dimethyl-1-oxo-2,7-octadienyl]-[(ß-D-glucopyranosyl)oxy]-2,6-dimethyl-1-oxo-2,7-octadienyl]-[(ß-D-glucopyranosyl)oxy]-2,6-dimethyl-1-oxo-2,7-octadienyl]-[(ß-D-glucopyranosyl)oxy]-2-(hydroxymethyl)-6-methyl-1-oxo-2,7-octadienyl]oxy]-16-hydroxy-3-[[O-ß-D-xylopyranosyl-(1 → 2)-O-α-L-arabinopyranosyl-(1 → 6)-2-(acetylamino)-2-deoxy-ß-D-glucopyranosyl]oxy]-(3ß,16α,21ß)-olean-12-en-28-oic acid O-α-L-arabinofuranosyl-(1 → 4)-O-[ß-D-glucopyranosyl-(1 → 3)]-O-6-deoxy-α-L-mannopyranosyl-(1 → 2)-ß-D-glucopyranosyl ester (1). The hemolytic potential of 1 was evaluated using in vitro assays, and its adjuvant activity on the cellular immune response against ovalbumin antigen was investigated using in vivo models.

Early postnatal, but not late, exposure to chemical ambient pollutant 1,2-naphthoquinone increases susceptibility to pulmonary allergic inflammation at adulthood.

High diesel exhaust particle levels are associated with increased health effects; however, knowledge on the impact of its chemical contaminant 1,2-naphthoquinone (1,2-NQ) is limited. We investigated whether postnatal and adult exposures to 1,2-NQ influence allergic reaction and the roles of innate and adaptive immunity. Male neonate (6 days) and adult (56 days) C57Bl/6 mice were exposed to 1,2-NQ (100 nM; 15 min) for 3 days, and on day 59, they were sensitized and later challenged with ovalbumin (OVA). Airway hyper-responsiveness (AHR) and production of cytokines, immunoglobulin E (IgE) and leukotriene B4 (LTB4) were measured in the airways. Postnatal exposure to 1,2-NQ activated dendritic cells in splenocytes by increasing expressing cell surface molecules (e.g., CD11c). Co-exposure to OVA effectively polarized T helper (Th) type 2 (Th2) by secreting Th2-mediated cytokines. Re-stimulation with unspecific stimuli (PMA and ionomycin) generated a mixed Th1 (CD4(+)/IFN-γ(+)) and Th17 (CD4(+)/IL-17(+)) phenotype in comparison with the vehicle-matched group. Postnatal exposure to 1,2-NQ did not induce eosinophilia in the airways at adulthood, although it evoked neutrophilia and exacerbated OVA-induced eosinophilia, Th2 cytokines, IgE and LTB4 production without affecting AHR and mast cell degranulation. At adulthood, 1,2-NQ exposure evoked neutrophilia and increased Th1/Th2 cytokine levels, but failed to affect OVA-induced eosinophilia. In conclusion, postnatal exposure to 1,2-NQ increases the susceptibility to antigen-induced asthma. The mechanism appears to be dependent on increased expression of co-stimulatory molecules, which leads to cell presentation amplification, Th2 polarization and enhanced LTB4, humoral response and Th1/Th2 cytokines. These findings may be useful for future investigations on treatments focused on pulmonary illnesses observed in children living in heavy polluted areas.