Hypoglycemic effect of Coffea arabica leaf extracts and major bioactive constituents.

The present study focused on investigating the antioxidant, antiglycation activity, digestive enzymes inhibition, bioaccessibility and hypoglycemic effect of C. arabica leaves extracts. The extracts deactivated the O2•-, ROO•, H2O2, HOCl reactive oxygen species. Coffee leaves showed strong inhibition of α-glucosidase (IC50 = 40.30 µg mL-1) greater than the isolated metabolites and acarbose. There was also inhibition of pancreatic lipase (IC50 = 56.43 µg mL-1) in addition to a hypoglycemic effect in zebrafish similar to acarbose and metformin. With the exception of rutin, all biocompounds were detected at all stages of in vitro digestion. Finally, these results suggest that C. arabica leaf extracts possess antidiabetic and anti-obesity properties that can be attributed to the main metabolites and the synergistic action between them.Communicated by Ramaswamy H. Sarma.

Extracellular matrix in skin diseases: The road to new therapies.

BACKGROUND: The extracellular matrix (ECM) is a vital structure with a dynamic and complex organization that plays an essential role in tissue homeostasis. In the skin, the ECM is arranged into two types of compartments: interstitial dermal matrix and basement membrane (BM). All evidence in the literature supports the notion that direct dysregulation of the composition, abundance or structure of one of these types of ECM, or indirect modifications in proteins that interact with them is linked to a wide range of human skin pathologies, including hereditary, autoimmune, and neoplastic diseases. Even though the ECM's key role in these pathologies has been widely documented, its potential as a therapeutic target has been overlooked. AIM OF REVIEW: This review discusses the molecular mechanisms involved in three groups of skin ECM-related diseases - genetic, autoimmune, and neoplastic - and the recent therapeutic progress and opportunities targeting ECM. KEY SCIENTIFIC CONCEPTS OF REVIEW: This article describes the implications of alterations in ECM components and in BM-associated molecules that are determinant for guaranteeing its function in different skin disorders. Also, ongoing clinical trials on ECM-targeted therapies are discussed together with future opportunities that may open new avenues for treating ECM-associated skin diseases.

Metals and low dose IR: Molecular effects of combined exposures using HepG2 cells as a biological model.

Uranium mining sites produce residues rich in metals and radionuclides, that may contaminate all environmental matrices, exposing human and non-human biota to low doses of ionizing radiation (LDIR) and to the chemical toxicity of several metals. To date, experimental and radio-epidemiological studies do not provide conclusive evidence of LDIR induced cancer. However, co-exposures (LDIR plus other contaminants), may increase the risks. To determine the potential for genotoxic effects in human cells induced by the exposure to LDIR plus metals, HEPG2 cells were exposed to different concentrations of a uranium mine effluent for 96 h. DNA damage was evaluated using the comet assay and changes in the expression of tumor suppressor and oncogenes were determined using qPCR. Results show that effluent concentrations higher than 5%, induce significant DNA damage. Also, a significant under-expression of ATM and TP53 genes and a significant overexpression of GADD45a gene was observed. Results show that the exposure to complex mixtures cannot be disregarded, as effects were detected at very low doses. This study highlights the need for further studies to clarify the risks of exposure to LDIR along with other stressors, to fully review the IR exposure risk limits established for human and non-human biota.

Molecular analysis of the RHD pseudogene by duplex real-time polymerase chain reaction.

OBJECTIVES: In this study, we aimed to present a strategy for the detection of the RHD pseudogene (RHDψ) based on a real-time polymerase chain reaction (PCR) assay. BACKGROUND: The D-negative phenotype is associated with many genetic alterations. In populations with African ancestry, this phenotype commonly results from the silent variant RHDψ. The evaluation of RHDψ is essential for correct inference of the RhD phenotype in order to avoid false-positive results. METHODS: We utilised a new method for the simultaneous detection of RHDψ and a fragment from exon 5 of the wild-type RHD gene based on duplex real-time PCR assay. RESULTS: The PCR assay allowed specific detection of RHDψ. There was complete agreement between the results generated by the new test and the results generated by molecular analysis performed using end-point PCR methods previously described. CONCLUSIONS: The assay developed is easy to execute and presents the potential for routine use at blood banks and other associated facilities where it is desired to determine the presence of RHDψ.

Life history traits and genotoxic effects on Daphnia magna exposed to waterborne uranium and to a uranium mine effluent - A transgenerational study.

Assessing the impact of uranium mining industry on aquatic ecosystems near mining areas is critical to ensure the long-term health and sustainability of ecosystem services. As so, a transgenerational study with Daphnia magna has been conducted to perceive to what extent intermittent discharges of uranium mine effluents into watercourses may impact the DNA integrity and life history traits of cladocerans. Organisms were exposed for 48 h to a 2% dilution of an uranium mine effluent (UME) and to a corresponding dose of waterborne uranium (WU) that, according to our preliminary studies, induces significant DNA damage in daphnids. After exposure, organisms were transferred to clean medium, where three successive generations were monitored for genotoxicity and other effects at the individual and population level. Despite some differences between WU and UME data, our results revealed that the negative impacts of the short-term exposure gradually disappeared after placing the organisms in clean medium. These results suggest that, under intermittent stress, daphnids are able to recover, since after the 3rd brood release, DNA damage (measured as DNA strand breaks) is no longer observed and has no significant impact on the detectable life traits of offspring. Although our results indicate that populations of D. magna are not affected by intermittent and highly diluted discharges from uranium mining, aquatic systems under this kind of pressure should not be seen as hazardous-free. Future studies in this field are recommended and these should consider radionuclides in the water column, their accumulation in the sediments and also multiple life stages.

RIBE at an inter-organismic level: A study on genotoxic effects in Daphnia magna exposed to waterborne uranium and a uranium mine effluent.

The induction of RIBE (Radiation Induced Bystander Effect) is a non-target effect of low radiation doses that has already been verified at an inter-organismic level in fish and small mammals. Although the theoretical impact in the field of environmental risk assessment (ERA) is possible, there is a gap of knowledge regarding this phenomenon in invertebrate groups and following environmentally relevant exposures. To understand if RIBE should be considered for ERA of radionuclide-rich wastewaters, we exposed Daphnia magna (<24 h and 5d old) to a 2% diluted uranium mine effluent for 48 h, and to a matching dose of waterborne uranium (55.3 µg L-1). Then the exposed organisms were placed (24 and 48 h) in a clean medium together with non-exposed neonates. The DNA damage observed for the non-exposed organisms was statistically significant after the 24 h cohabitation for both uranium (neonates p = 0.002; 5 d-old daphnids p = <0.001) and uranium mine effluent exposure (only for neonates p = 0.042). After 48 h cohabitation significant results were obtained only for uranium exposure (neonates p = 0.017; 5 d-old daphnids p = 0.013). Although there may be some variability associated to age and exposure duration, the significant DNA damage detected in non-exposed organisms clearly reveals the occurrence of RIBE in D. magna. The data obtained and here presented are a valuable contribution for the discussion about the relevance of RIBE for environmental risk assessment.

Source identification of uranium-containing materials at mine legacy sites in Portugal.

Whilst prior nuclear forensic studies have focused on identifying signatures to distinguish between different uranium deposit types, this paper focuses on providing a scientific basis for source identification of materials from different uranium mine sites within a single region, which can then be potentially used within nuclear forensics. A number of different tools, including gamma spectrometry, alpha spectrometry, mineralogy and major and minor elemental analysis, have been utilised to determine the provenance of uranium mineral samples collected at eight mine sites, located within three different uranium provinces, in Portugal. A radiation survey was initially conducted by foot and/or unmanned aerial vehicle at each site to assist sample collection. The results from each mine site were then compared to determine if individual mine sites could be distinguished based on characteristic elemental and isotopic signatures. Gamma and alpha spectrometry were used to differentiate between samples from different sites and also give an indication of past milling and mining activities. Ore samples from the different mine sites were found to be very similar in terms of gangue and uranium mineralogy. However, rarer minerals or specific impurity elements, such as calcium and copper, did permit some separation of the sites examined. In addition, classification rates using linear discriminant analysis were comparable to those in the literature.

Human leukocyte antigen distribution and genomic ancestry in Brazilian patients with sickle cell disease.

Hematopoietic stem-cell transplantation (HSCT) is currently the only established curative treatment for sickle cell disease (SCD), but is limited by donor availability. Ethnicity is thought to have an impact on the complications experienced by patients that undergo HSCT and on the likelihood of identifying an human leukocyte antigen (HLA) matched donor. In the present study, we investigated the genomic ancestry and the distribution of HLA allele groups in Brazilian patients with SCD, compared these HLA profiles to worldwide populations and evaluate the availability of HLA-matched donors. A broad intercontinental admixture of patients with SCD was observed, with African ancestry ranging from 6.7% to 93.4%. In a dendrogram based on HLA frequencies, Brazilian patients with SCD were included in a branch containing only populations with a significant African component. Among the 126 patients evaluated, 10 (8%) found a HLA-matched unrelated donor in a database of 18 134 donors. Self-reported white, brown and black matched donors were identified, and no significant difference in the percentage of compatible donors was observed between these ethnic groups. Our results show that Brazilian patients with SCD are very admixed, indicating that this group is a promising target for admixture mapping of genes involved in complications after HSCT. Additional studies may help to clarify the impact of the genetic diversity and admixture of these patients on the donor availability.

Uranium mining wastes: The use of the Fish Embryo Acute Toxicity Test (FET) test to evaluate toxicity and risk of environmental discharge.

Active and abandoned uranium mining sites often create environmentally problematic situations, since they cause the contamination of all environmental matrices (air, soil and water) with stable metals and radionuclides. Due to their cytotoxic, genotoxic and teratogenic properties, the exposure to these contaminants may cause several harmful effects in living organisms. The Fish Embryo Acute Toxicity Test (FET) test was employed to evaluate the genotoxic and teratogenic potential of mine liquid effluents and sludge elutriates from a deactivated uranium mine. The aims were: a) to determine the risk of discharge of such wastes in the environment; b) the effectiveness of the chemical treatment applied to the uranium mine water, which is a standard procedure generally applied to liquid effluents from uranium mines and mills, to reduce its toxicological potential; c) the suitability of the FET test for the evaluation the toxicity of such wastes and the added value of including the evaluation of genotoxicity. Results showed that through the FET test it was possible to determine that both elutriates and effluents are genotoxic and also that the mine effluent is teratogenic at low concentrations. Additionally, liquid effluents and sludge elutriates affect other parameters namely, growth and hatching and that water pH alone played an important role in the hatching process. The inclusion of genotoxicity evaluation in the FET test was crucial to prevent the underestimation of the risks posed by some of the tested effluents/elutriates. Finally, it was possible to conclude that care should be taken when using benchmark values calculated for specific stressors to evaluate the risk posed by uranium mining wastes to freshwater ecosystems, due to their chemical complexity.

Effect of red blood cell preservation by droplet freezing with non-permeable cryoprotective agents in blood group antigen reactivity.

BACKGROUND: In the last few decades, various red blood cell (RBC) freezing techniques have been developed and improved to enable the preservation of erythrocytes for future use in pre-transfusion tests in reference immunohaematology laboratories. However, not all these techniques have been sufficiently evaluated for the preservation of blood group antigens. OBJECTIVES: In this study, we evaluated the antigenic pattern of RBCs preserved by droplet freezing in liquid nitrogen in a blood bank context. METHODS: Blood samples were evaluated for the reactivity of blood group antigens after droplet freezing using the non-permeable cryoprotective agent polyvinylpyrrolidone (PVP) and sucrose-dextrose (S + D) solutions. RESULTS: No qualitative changes were observed in RBC reactivity after freezing and thawing for the antigens Fyb , Leb , C, E, Cw , Lua , Lub , Kpa , Kpb and Dia . However, cryopreservation using PVP resulted in a significant increase in reactivity of Fyb antigen on comparing fresh and frozen samples (P < 0·001). CONCLUSION: The establishment of detailed protocols for cryopreservation of RBCs, which take into account the maintenance of antigenic characteristics, is necessary to increase security in pre-transfusion testing using frozen RBCs.

Tuberculosis Caused by Mycobacterium bovis in a Capybara (Hydrochoerus hydrochaeris).

Tuberculosis, associated with Mycobacterium bovis, was diagnosed post mortem in an adult female capybara (Hydrochoerus hydrochaeris), kept at the Pampulha Ecological Park, Belo Horizonte, Brazil, in a large metropolitan area. On post-mortem examination, there were numerous firm white nodules scattered throughout all lobes of both lungs. Tissue samples were collected for histological and microbiological examination. Microscopically, the pulmonary nodules were multifocal to coalescing granulomas and intralesional acid-fast bacilli were evident in Ziehl-Neelsen-stained sections of the lung and spleen. Colonies with morphological features of Mycobacterium spp. were isolated from lung samples and conventional polymerase chain reaction (PCR) with genomic DNA from the isolates was positive for M. bovis; sequencing indicated 100% identity with the region of difference 4 (RD4) of M. bovis. In addition, M. bovis DNA was detected in the lung by quantitative PCR. The finding of M. bovis in a capybara indicates a potential public health risk in a zoological collection.

Dried blood spots of pooled samples for RHD gene screening in blood donors of mixed ancestry.

OBJECTIVES: In this study, we present a strategy for RHD gene screening based on real-time polymerase chain reaction (PCR) using dried blood spots of pooled samples. BACKGROUND: Molecular analysis of blood donors may be used to detect RHD variants among the presumed D-negative individuals. RHD genotyping using pooled samples is a strategy to test a large number of samples at a more reasonable cost. MATERIALS AND METHODS: RHD gene detection based on real-time PCR using dried blood spots of pooled samples was standardised and used to evaluate 1550 Brazilian blood donors phenotyped as RhD-negative. Positive results were re-evaluated by retesting single samples using real-time PCR and conventional multiplex PCR to amplify five RHD-specific exons. PCR-sequence-specific primers was used to amplify RHDψ allele. RESULTS: We devised a strategy for RHD gene screening using dried blood spots of five pooled samples. Among 1550 serologically D-negative blood donors, 58 (3.74%) had the RHD gene. The non-functional RHDψ allele was detected in 47 samples (3.02%). CONCLUSION: The present method is a promising strategy to detect the RHD gene among presumed RhD-negative blood donors, particularly for populations with African ancestry.

The local environment of Co2+ ions intercalated in vanadium oxide/hexadecylamine nanotubes.

Vanadium oxide nanotubes constitute promising materials for applications in nanoelectronics as cathode materials, in sensor technology and in catalysis. In this work we present a study on hybrid vanadium oxide/hexadecylamine multiwall nanotubes doped with Co ions using state of the art x-ray diffraction and absorption techniques, to address the issue of the dopant location within the nanotubes' structure. The x-ray absorption near-edge structure analysis shows that the Co ions in the nanotubes are in the 2 + oxidation state, while extended x-ray absorption fine structure spectroscopy reveals the local environment of the Co(2+) ions. Results indicate that Co atoms are exchanged at the interface between the vanadium oxide's layers and the hexadecylamines, reducing the amount of amine chains and therefore the interlayer distance, but preserving the tubular shape. The findings in this work are important for describing Co(2+) interaction with vanadium oxide nanotubes at the molecular level and will help to improve the understanding of their physicochemical behavior, which is desired in view of their promising applications.

Radioactivity from Fukushima nuclear accident detected in Lisbon, Portugal.

The radioactivity released from the Fukushima Dai-ichi nuclear accident was transported around the globe by atmospheric processes. Several artificial radionuclides were detected and measured in aerosols and atmospheric surface depositions in the Lisbon area during late March and early April 2011. The highest concentrations measured in aerosols were those of particulate (131)I, 1.39 ± 0.08 mBq m(-3). Cesium-134, (137)Cs and (132)Te were also determined but at lower concentrations. The total atmospheric depositions on the ground were higher on the first week of April with values for (131)I, (134)Cs, and (137)Cs of 0.92 ± 0.11, 0.59 ± 0.06, and 0.62 ± 0.12 Bq m(-2), respectively. The four artificial radionuclides measurable, (131)I, (132)Te (134)Cs, and (137)Cs, caused little radiation exposure to the members of the public, that was five orders of magnitude lower than the ionizing radiation effective dose limits for members of the public for one year (1 mSv y(-1)).

Discussing childbearing with HIV-infected women of reproductive age in clinical care: a comparison of Brazil and the US.

Despite long term access to highly active antiretroviral therapy in Brazil and the US, little is known about women's communication with their HIV provider regarding childbearing or the unmet need for reproductive counseling. We utilized identical survey questions to collect data from HIV-infected women of reproductive age in Rio de Janeiro (n = 180) and Baltimore (n = 181). We conducted univariate analyses to compare findings between samples of women and multivariate logistic regression to determine factors associated with childbearing desires, childbearing intentions, and provider communication among the combined sample of women (n = 361). Over one-third of women in Rio de Janeiro and nearly one-half of women in Baltimore reported the desire for future childbearing. Nevertheless, the majority of women in clinical care had not discussed future childbearing with their HIV provider. Even in countries with an advanced approach to HIV care, we found low and inadequate communication between providers and female patients about childbearing.

Tubulopapillary carcinoma of the mammary gland in a maned wolf (Chrysocyon brachyurus): histopathological and immunophenotypical analysis / Carcinoma túbulo-papilar da glândula mamária em lobo guará fêmea (Chrysocyon brachyurus): análise histopatológica e imunofenotípica

A maned female wolf (Chrysocyon brachyurus) showed nodules in the inguinal and left abdominal cranial mammary glands. The mammary gland was surgically excised, and microscopic analysis revealed epithelial cell proliferation in a tubular and papillary pattern; delicate fibrovascular stalks presenting numerous layers of moderately pleomorfic epithelial cells were observed. This histologic appearance was compatible with a diagnosis of mammary tubulopapillary carcinoma. The immunohistochemical profile revealed nuclear positivity for estrogen (70 percent) and progesterone (at least 90 percent of the neoplastic cells). The myoepithelium-associated with neoplastic cells lacked integrity, as evidenced by failed smooth muscle alpha actin reactivity in microinvasive areas. A low proliferation index was observed (3.4 percent). To the authors' knowledge, the present case represents the first finding of female tubulopapillary carcinoma in a mammary gland in this species.

Um lobo guará fêmea (Chrysocyon brachyurus) apresentou nódulos nas mamas inguinal e abdominal cranial esquerda. As glândulas mamárias foram cirurgicamente excisadas, e na análise microscópica revelaram proliferação de células epiteliais em arranjo tubular e papilar com tecido conjuntivo de sustentação delicado e formação múltiplas camadas de células epiteliais moderadamente pleomórficas. Os achados histopatológicos foram compatíveis com carcinoma túbulo-papilar da glândula mamária. O estudo do perfil imuno-histoquímico mostrou positividade nuclear para estrógeno (70 por cento) e progesterona (em pelo menos 90 por cento das células). Observou-se perda da integridade da camada mioepitelial, adjacente a proliferação epitelial, evidenciada pela perda de reatividade para alfa actina de músculo liso em áreas de microinvasão. Índice de proliferação celular baixo foi observado (3,4 por cento). O presente relato representa a primeira descrição de um carcinoma túbulo-papilar da glândula mamária em uma fêmea desta espécie.

Cutaneous angiomatosis in a llama (Lama glama).

Cutaneous angiomatosis was diagnosed in an adult female llama (Lama glama). Lesions were raised or plaque-like, erythematous, firm to soft in consistency and were observed on the face and skin of the axillary, abdominal, perineal and inguinal regions. The lesions were not painful or pruritic. Microscopical examination revealed an irregular parakeratotic lamellar hyperkeratosis associated with diffuse proliferation of arterioles and venules in the superficial dermis. Immunohistochemical analysis determined that the cells forming these vessels and perivascular cells expressed factor VIII-related antigen, vascular endothelial growth factor (VEGF), CD31 and smooth-muscle alpha-actin. These studies confirmed the diagnosis of cutaneous angiomatosis.

Comparison of serum hormone levels of captive and free-living maned wolves Chrysocyon brachyurus.

Serum hormone levels were compared between captive and free-living maned wolves and seasonal variations of sex hormones were studied. Blood samples were collected from 16 male and 26 female adult animals from Brazilian zoos, and from 30 male and 24 female free-living adults to determine serum progesterone and testosterone by radioimmunoassay. Serum testosterone concentrations varied (P < 0.05) across seasons for 16 captive males, being higher in autumn (2184.7 +/- 355.1 pg/mL) than in summer (1080.7 +/- 205.4 pg/mL), winter (1270.1 +/- 276.6 pg/mL) and spring (963.9 +/- 248.1 pg/mL), although they did not differ between summer, winter and spring. Testosterone concentration of 30 free-living males differed (P < 0.05) between autumn (824.1 +/- 512.2 pg/mL), winter (14.4 +/- 8.0 pg/mL) and spring (151.9 +/- 90.5 pg/mL). Comparison between captive and free-living animals showed no difference in autumn (P > 0.05). Sixteen captive males showed higher testosterone concentration during winter and spring compared with 30 free-living animals (P < 0.05). Progesterone concentration varied among seasons in 26 captive females (P < 0.05), being higher in autumn (15.3 +/- 3.1 ng/mL) than in summer (6.6 +/- 1.5 ng/mL), winter (5.3 +/- 3.1 ng/mL) and spring (4.3 +/- 0.7 ng/mL). Progesterone concentration of 24 free-living females varied between autumn (17.1 +/- 6.0 ng/mL) and winter (1.7 +/- 0.3 ng/mL) (P < 0.05), but we could not obtain data for spring or summer. No difference in progesterone levels was observed between captive and free-living females in autumn and winter.

Comparison of serum hormone levels of captive and free-living maned wolves Chrysocyon brachyurus

Serum hormone levels were compared between captive and free-living maned wolves and seasonal variations of sex hormones were studied. Blood samples were collected from 16 male and 26 female adult animals from Brazilian zoos, and from 30 male and 24 female free-living adults to determine serum progesterone and testosterone by radioimmunoassay. Serum testosterone concentrations varied (P < 0.05) across seasons for 16 captive males, being higher in autumn (2184.7 ± 355.1 pg/mL) than in summer (1080.7 ± 205.4 pg/mL), winter (1270.1 ± 276.6 pg/mL) and spring (963.9 ± 248.1 pg/mL), although they did not differ between summer, winter and spring. Testosterone concentration of 30 free-living males differed (P < 0.05) between autumn (824.1 ± 512.2 pg/mL), winter (14.4 ± 8.0 pg/mL) and spring (151.9 ± 90.5 pg/mL). Comparison between captive and free-living animals showed no difference in autumn (P > 0.05). Sixteen captive males showed higher testosterone concentration during winter and spring compared with 30 free-living animals (P < 0.05). Progesterone concentration varied among seasons in 26 captive females (P < 0.05), being higher in autumn (15.3 ± 3.1 ng/mL) than in summer (6.6 ± 1.5 ng/mL), winter (5.3 ± 3.1 ng/mL) and spring (4.3 ± 0.7 ng/mL). Progesterone concentration of 24 free-living females varied between autumn (17.1 ± 6.0 ng/mL) and winter (1.7 ± 0.3 ng/mL) (P < 0.05), but we could not obtain data for spring or summer. No difference in progesterone levels was observed between captive and free-living females in autumn and winter.

An outbreak of sarcocystosis in psittacines and a pigeon in a zoological collection in Brazil.

This report describes an outbreak of acute pulmonary sarcocystosis in different species of captive psittacines and in a Luzon bleeding-heart pigeon (Gallicolumba luzonica) in a zoological collection in Brazil. A majority of the birds were found dead and had exhibited no previous clinical signs. Grossly, pulmonary congestion and edema were the most-common findings. Enlarged and congested livers and spleens were also frequently observed. Microscopically, there was edema, fibrin exudation, congestion, and perivascular and interstitial lymphoplasmacytic infiltration associated with numerous sinuous schizonts of Sarcocystis sp. in the lungs. Mild to moderate myocarditis, hepatitis, splenitis, and interstitial nephritis were also observed in the birds. Immunohistochemistry confirmed Sarcocystis sp. in the capillaries of lungs, hearts, livers, and spleens of most of the birds, but also in the pancreas, kidney, intestine, proventriculus, and brain of a few birds. The probable source of Sarcocystis sp. in these birds was the wild opossum (Didelphis albiventris), a common inhabitant of a local forest that surrounds the Belo Horizonte Zoo (Fundação Zoo-Botânica). This is the first documentation of Sarcocystis infection in psittacines and a pigeon from Brazil.