Polarization of Melatonin-Modulated Colostrum Macrophages in the Presence of Breast Tumor Cell Lines.

Human colostrum and milk contain diverse cells and soluble components that have the potential to act against tumors. In breast cancer, macrophages play a significant role in immune infiltration and contribute to the progression and spread of tumors. However, studies suggest that these cells can be reprogrammed to act as an antitumor immune response. This study aimed to evaluate the levels of melatonin and its receptors, MT1 (melatonin receptor 1) and MT2 (melatonin receptor 2), in colostrum and assess the differentiation and polarization of the colostrum macrophages modulated by melatonin in the presence of breast tumor cells. Colostrum samples were collected from 116 mothers and tested for their melatonin and receptor levels. The colostrum cells were treated with or without melatonin and then cultured for 24 h in the presence or absence of breast tumor cells. The results showed that melatonin treatment increased the expression of MT1 and MT2 in the colostrum cells. Furthermore, melatonin treatment increased the percentage of M1 macrophages and decreased the percentage of M2 macrophages. When the colostrum macrophages were cocultured with breast tumor cells, melatonin reduced the percentage of both macrophage phenotypes and the cytokines tumor necrosis factor-alpha (TNF-α) and interleukin 8 (IL-8). These data suggest that melatonin can regulate the inflammatory process via M1 macrophages in the tumor microenvironment and, simultaneously, the progression of M2 macrophages that favor tumorigenesis.

Bioactive compounds of pequi pulp and oil extracts modulate antioxidant activity and antiproliferative activity in cocultured blood mononuclear cells and breast cancer cells.

BACKGROUND: Pequi (Caryocar brasiliense Camb.) is a fruit from Brazilian Cerrado rich in bioactive compounds, such as phytosterols and tocopherols, which can modulate the death of cancer cells. OBJECTIVE: In the present study, the main bioactive compounds of hydrophilic and lipophilic extracts of pequi oil and pulp were identified and were verified if they exert modulatory effects on oxidative stress of mononuclear cells cocultured with MCF-7 breast cancer cells. STUDY DESIGN: Identification and quantification of the main compounds and classes of bioactive compounds in pequi pulp and oil, hydrophilic, and lipophilic extracts were performed using spectroscopy and liquid chromatographic methods, while the beneficial effects, such as antioxidant capacity in vitro, were determined using methods based on single electron transfer reaction or hydrogen atom transfer, while for antioxidant and antiproliferative activities ex vivo, 20 healthy volunteers were recruited. Human peripheral blood mononuclear cells (MN) were collected, and cellular viability assay by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide), superoxide anion evaluation, and CuZn-superoxide dismutase determination (CuZn-SOD) in MN cells, MCF-7 cells, and coculture of MN cells and MCF-7 cells in the presence and absence of pequi pulp or oil hydrophilic and lipophilic extracts were performed. RESULTS: In the hydrophilic extract, the pequi pulp presented the highest phenolic content, while in the oil lipophilic extract, it had the highest content of carotenoids. The main phytosterol in pequi oil was ß-sitosterol (10.22 mg/g), and the main tocopherol was γ-tocopherol (26.24 µg/g sample). The extracts that had highest content of bioactive compounds stimulated blood mononuclear cells and also improved SOD activity. By evaluating the extracts against MCF-7 cells and coculture, they showed cytotoxic activity. CONCLUSION: The results support the anticarcinogenic activity of pequi extracts, in which the pequi pulp hydrophilic extracts presented better immunomodulatory potential.

Interferon-gamma carrying nanoemulsion with immunomodulatory and anti-tumor activities.

The therapeutic administration of cytokines has been introduced aiming to modulate the immune response system, seeking for different approaches to face pathologies such as cancer, auto immune and infectious diseases. The objective of this study was to investigate the effects of a stable oil-in-water (O/W) nanoemulsion system carrying the cytokine Interferon gamma (IFN-γ) on the activity of phagocytes and MCF-7 human breast cancer cells. Nanoemulsions were prepared through ultra-homogenization, and they consisted of distilled water, triglycerides of capric acid/caprylic, sorbitan-oleate, polysorbate 80, and 1-butanol. IFN-γ (100 ng ml-1 ) was incorporated into two O/W nanoemulsion formulations, and these formulations were characterized in terms of their preliminary and accelerated physicochemical stability, rheological properties, droplet size, polydispersity and surface charge. We identified the most optimal IFN-γ nanoemulsion (IFN-γNE2), which remained stable under extreme temperature variations for 90 days, contained an average dose of 97 ng ml-1 of IFN-γ and exhibited a biocompatible pH and a relative stable rheological profile. Cell viability and intracellular Ca2+ release assays conducted showed that IFN-γNE2 reduced the cell viability of MCF-7 cells without affecting the cell viability of phagocytes. Furthermore, IFN-γNE2 was able to induce cellular activity of phagocytes as evidenced by increased intracellular Ca2+ release in these cells. Our findings on this IFN-γ nanoemulsion suggest that it can be a promising therapeutic agent for immunostimulation and cancer treatment.

O-Glycosylation with O-linked ß-N-acetylglucosamine increases vascular contraction: Possible modulatory role on Interleukin-10 signaling pathway.

AIMS: The interleukin-10 (IL-10) is an immuno-regulatory cytokine that plays a protective effect in the vasculature. IL-10 binding to its receptor, activating the IL-10/JAK1/STAT3 cascade to exert its effects. Therefore, STAT3 phosphorylation is essential for IL-10 actions. O-Glycosylation with linked ß-N-acetylglucosamine (O-GlcNAc) is a post-translational modification able to regulate many proteins by interfering with protein on a phosphorylation level. Our aim was to determine whether O-GlcNAc promotes the inhibition of IL-10-pathway (JAK1/STAT3/IL-10), inactivationg its action in the vasculature. MAIN METHODS: Mice (C57BL/6) aortic segments were incubated with vehicle or Thiamet G (0.1 mM, for 24 h) to increase global O-GlcNAc levels. Aortas from knockout mice for IL-10 were also used. Vascular reactivity and western blot tests were performed to evaluate protein expression. KEY FINDINGS: High levels of O-GlcNAc, induced by Thiamet G incubation, increased vascular expression of JAK1, but decreased expression and activity of STAT3. In addition, IL-10 levels were diminished in arteries treated with Thiamet G. Absence of IL-10, as well as augmented O-GlcNAcylation, increased vascular reactivity to constrictor stimuli, an effect that was abolished by ERK 1/2 inhibitor. High levels of O-GlcNAc and the absence of IL-10 also leads to increased vascular expression of ERK1/2. SIGNIFICANCE: Our data suggest that O-GlcNAc modification seems to (dys)regulate IL-10 signaling pathway and consequently, compromise the protective effect of this cytokine in vasculature. It is possible that there is a promising relationship in pathophysiological conditions where changes in O-GlcNAcylation and IL-10 levels are observed, such as hypertension and diabetes.

Immunohaematological and rheological parameters in canine visceral leishmaniasis / Parâmetros reológicos e imunohematológicos na leishmaniose visceral canina

Abstract The aim of this study was to find correlations between haematological and rheological parameters in canine visceral leishmaniasis (CLV). Enrolled dogs with CVL (n=31) had a confirmed diagnosis using the rapid test Dual Path Platform (TR DPP®) kit and Polymerase Chain Reaction (PCR) assay. A control group (n=31) comprised healthy dogs with negative results for the TR DPP® kit and PCR assay. Haematological parameters and total protein, albumin, globulin, and antibody were assessed. The rheological properties of the blood samples were also determined. Erythrocytes, haematocrit and platelet values of dogs in the CVL group were found to be lower than those of the control group. The total protein, globulin and IgG concentrations were higher in serum samples from the CVL group. The blood flow curve demonstrated increased shear rates in the CVL group. The viscosity of the blood from the infected animals was lower than in the healthy dogs. A positive correlation was found between erythrocyte numbers and blood viscosity. These data suggest that changes in the flow curve and viscosity of blood as well as the erythrocyte values may be a viable low-cost alternative for the monitoring of dogs with visceral leishmaniasis.

Resumo O objetivo deste estudo foi encontrar correlações entre parâmetros hematológicos e reológicos na leishmaniose visceral canina (LVC). Cães com LVC (n=31) tiveram diagnóstico confirmado de LVC pelo teste rápido imunocromatográfico (TR DPP®) e o ensaio de Reação em Cadeia da Polimerase (PCR). Cães saudáveis ​​ negativos para o TR DPP® e PCR foram alocados no grupo controle (n=31). Foram avaliados parâmetros hematológicos, proteínas totais, albumina, globulina, anticorpos e as propriedades reológicas do sangue. Os valores de eritrócitos, hematócrito e plaquetas de cães do grupo LVC foram menores do que os encontrados no grupo controle. As concentrações de proteína total, globulina e imunoglobulina G (IgG) foram maiores em amostras de soro do grupo LVC. A curva do fluxo sanguíneo apresentou maior taxa de cisalhamento no grupo LVC. A viscosidade sanguínea dos animais infectados foi menor do que a viscosidade do sangue dos animais do grupo controle. Uma correlação positiva foi encontrada entre o número de eritrócitos e a viscosidade do sangue. Esses dados sugerem que as mudanças na curva de fluxo e viscosidade do sangue, bem como os valores de eritrócitos, podem ser uma alternativa viável e de baixo custo para o monitoramento de cães com leishmaniose visceral.

Immunohaematological and rheological parameters in canine visceral leishmaniasis.

The aim of this study was to find correlations between haematological and rheological parameters in canine visceral leishmaniasis (CLV). Enrolled dogs with CVL (n=31) had a confirmed diagnosis using the rapid test Dual Path Platform (TR DPP®) kit and Polymerase Chain Reaction (PCR) assay. A control group (n=31) comprised healthy dogs with negative results for the TR DPP® kit and PCR assay. Haematological parameters and total protein, albumin, globulin, and antibody were assessed. The rheological properties of the blood samples were also determined. Erythrocytes, haematocrit and platelet values of dogs in the CVL group were found to be lower than those of the control group. The total protein, globulin and IgG concentrations were higher in serum samples from the CVL group. The blood flow curve demonstrated increased shear rates in the CVL group. The viscosity of the blood from the infected animals was lower than in the healthy dogs. A positive correlation was found between erythrocyte numbers and blood viscosity. These data suggest that changes in the flow curve and viscosity of blood as well as the erythrocyte values may be a viable low-cost alternative for the monitoring of dogs with visceral leishmaniasis.

Relationship between Proinflammatory Cytokines/Chemokines and Adipokines in Serum of Young Adults with Obesity.

BACKGROUND AND OBJECTIVE: The adipose tissue has been recognized as an important endocrine organ, which is metabolically active and expresses and secretes various inflammatory cytokines. Inflammation is involved in obesity-related complications. As such, the present study investigated the correlation between biochemical parameters, serum proinflammatory cytokines and adipokines in individuals with obesity. METHODS: Based on the body mass index (BMI), 30 subjects were divided into 3 groups: eutrophic (GC, n = 10), overweight (GOW, n = 10) and obese (GOB, n = 10). Serum glucose, cholesterol (total-C, HDLC and LDL-C), triglycerides, total proteins, uric acid and insulin were determined, as well as cytokines IL-8, TNF-α, IL-1ß, and IL-6, leptin and adiponectin. RESULTS: GOB showed the highest glucose, total and LDL-C, triglycerides, uric acid, insulin, leptin, IL- 8, IL-1ß, IL-6, TNF-α and lowest adiponectin levels. In general, adiponectin exhibited an inverse correlation with BMI, abdominal circumference, LDL-C, IL-6, TNF-α, leptin and leptin-adiponectin ratio (LAR) and a positive correlation with HDL-C. Leptin was positively correlated with BMI, abdominal circumference, insulin, IL-6, TNF-α and LAR and negatively correlated with HDL-C and adiponectin. The LAR was positively correlated with BMI, waist circumference, insulin, TNF-α and negatively associated with HDL-C. CONCLUSION: The results confirm that obesity changes the lipid and glycemic profiles of individuals, increases the proinflammatory adipokine levels and reduces those of anti-inflammatory adipokines, promoting a state of chronic inflammation.

Effects of barium chloride adsorbed to polyethylene glycol (PEG) microspheres on co-culture of human blood mononuclear cell and breast cancer cell lines (MCF-7).

This study investigated the effects of BaCl2 adsorbed to polyethylene glycol (PEG) microspheres on human blood mononuclear cells (MN) co-cultured with breast cancer cell lines (MCF-7). The MCF-7 cells were obtained from the American Type Culture Collection and the blood mononuclear (MN) cells from volunteer donors. MN cells, MCF-7 cells and their co-culture (MN and MCF-7 cells) were pre-incubated for 24 h with or without 25 and 1000 pg L-1 BaCl2 (Ba25 and Ba1000), PEG microspheres or 25 and 1000 pg L-1 BaCl2 adsorbed to PEG microspheres (PEG-Ba25 and PEG-Ba1000). Rheological parameters and apoptosis were determined. Fluorescence microscopy and flow cytometry analyses revealed that BaCl2 was able to adsorb the PEG microspheres. The blood flow and viscosity curves were similar among the treatments. In general, apoptosis rates increased in co-cultured cells, co-cultured cells incubated with Ba25 and with PEG-Ba25, but the highest rates were observed in co-cultured cells incubated with PEG-Ba1000. In conclusion, BaCl2 adsorbed to PEG microspheres exhibited dose-dependent antitumor effects against human MCF-7 breast cancer cells co-cultured with MN cells, thereby offering a possible therapeutic alternative for treating this disease provided they are administered at very low concentrations.

Herbal Mixture Adsorbed to Polyethylene Glycol Microspheres Induces Apoptotic Effects on Breast Cancer Cells.

BACKGROUND: The combination of medicinal plants and polymeric matrices raised the possibility of obtaining new drugs to treat a number of diseases, including cancer. Breast cancer is one of the most frequent diseases among women, but the effective treatments is still a challenge. METHODS: Thus, this study investigated the effect of herbal mixture adsorbed to PEG microspheres on MCF-7 human breast cancer cells and these cells in co-culture of blood MN cells. The MCF-7 cells and the blood mononuclear [MN] cells from volunteer donors. MN cells, MCF-7 cells and co-culture [MN and MCF-7 cells] were pre-incubated for 24 h with or without herbal mixture [HM], polyethylene glycol [PEG] microspheres or herbal mixture adsorbed in PEG microspheres [PEG-HM]. Viability, intracellular calcium and apoptosis were determined by flow cytometry. The herbal mixture adsorbed in PEG microspheres reduces the viability of MCF-7 cells. RESULTS: The lowest viability índex was observed in co-culture incubated with herbal mixture adsorbed to PEG microspheres. MN cells, MCF-7 cells and co-culture showed higher intracellular Ca2+ release when incubated with herbal mixture adsorbed to PEG microspheres. The apoptosis index was higher in MCF-7 cells that were treated with herbal mixture. The highest apoptosis index was observed in coculture of these cells incubated with herbal mixture adsorbed to PEG microspheres. CONCLUSION: These data suggest that herbal mixture adsorbed by PEG microspheres has apoptotic effects on human MCF-7 breast cancer cells and is therefore a possible therapeutic alternative.

Qualidade microbiológica da água tratada e não tratada da região de Ribeirão Preto, SP / Microbiological quality of treated and untreated water from the region of Ribeirao Preto, SP

O objetivo deste estudo foi avaliar a qualidade microbiológica de diferentes amostras de água tratada e não tratada provenientes da região de Ribeirão Preto/SP. Foram determinados coliformes totais e coliformes termotolerantes e contagem de micro-organismos heterotróficos aeróbios ou facultativos mesófilos viáveis presentes, verificando se as amostras analisadas atendem às normas da legislação vigente. Os resultados obtidos evidenciaram que as nove amostras de água tratada, atenderam à legislação quanto à potabilidade da água. Das 18 amostras de água não tratada, duas amostras (11,1 %) apresentaram valores superiores a 500 UFC/mL em relação à contagem de micro-organismos heterotróficos aeróbios ou facultativos mesófilos, além de apresentarem coliformes termotolerantes, sendo consideradas impróprias para o consumo humano, representando um risco na disseminação de doenças de veiculação hídrica.

Análise microbiológica de presunto cozido fatiado comercializado na cidade de Ribeirão Preto - SP / Microbiological analysis of sliced baked ham marketed in the city of Ribeirao Preto - SP

Objetivou-se neste trabalho analisar a qualidade microbiológica do presunto fatiado comercializado na cidade de Ribeirão Preto-SP. Os resultados das análises microbiológicas foram obtidos pela determinação de presença e contagem de micro- -organismos patógenos e/ou deteriorantes, dentre eles Salmonella, coliformes, clostrídios sulfito redutores, Staphylococcus aureus e mesófilos. Foram analisadas 30 amostras, sendo 15 fatiadas e embaladas à vácuo pelo próprio fabricante e 15 fatiadas e embaladas pelo estabelecimento comercial. As amostras diferiam em fabricante e estabelecimento de comercialização. Os dados evidenciam que 6,7% das amostras analisadas de presunto em embalagens à vácuo acusaram a presença de Staphylococcus aureus. Das amostras analisadas e embaladas pelo estabelecimento comercial 13,3% acusaram presença de Staphylococcus aureus e 33,3 % acusaram presença de coliformes. Os resultados obtidos indicam possíveis falhas nas condições higienicossanitárias durante a manipulação do produto.

Incidence of Y-chromosome microdeletions in children whose fathers underwent vasectomy reversal or in vitro fertilization with epididymal sperm aspiration: a case-control study / Incidência de microdeleções do cromossomo Y em filhos de pais que passaram por reversão de vasectomia ou fertilização in vitro com aspiração do epidídimo: um estudo caso-controle

ABSTRACT Objective To evaluate the incidence of Y-chromosome microdeletions in individuals born from vasectomized fathers who underwent vasectomy reversal or in vitro fertilization with sperm retrieval by epididymal aspiration (percutaneous epididymal sperm aspiration). Methods A case-control study comprising male children of couples in which the man had been previously vasectomized and chose vasectomy reversal (n=31) or in vitro fertilization with sperm retrieval by percutaneous epididymal sperm aspiration (n=30) to conceive new children, and a Control Group of male children of fertile men who had programmed vasectomies (n=60). Y-chromosome microdeletions research was performed by polymerase chain reaction on fathers and children, evaluating 20 regions of the chromosome. Results The results showed no Y-chromosome microdeletions in any of the studied subjects. The incidence of Y-chromosome microdeletions in individuals born from vasectomized fathers who underwent vasectomy reversal or in vitro fertilization with spermatozoa recovered by percutaneous epididymal sperm aspiration did not differ between the groups, and there was no difference between control subjects born from natural pregnancies or population incidence in fertile men. Conclusion We found no association considering microdeletions in the azoospermia factor region of the Y chromosome and assisted reproduction. We also found no correlation between these Y-chromosome microdeletions and vasectomies, which suggests that the assisted reproduction techniques do not increase the incidence of Y-chromosome microdeletions.

RESUMO Objetivo Avaliar a incidência de microdeleções do cromossomo Y em indivíduos nascidos de pais vasectomizados submetidos à reversão de vasectomia ou fertilização in vitro com recuperação de espermatozoides por aspiração do epidídimo (aspiração percutânea de espermatozoides do epidídimo). Métodos Estudo caso-controle que compreende crianças do sexo masculino de casais em que o homem havia sido previamente vasectomizado e escolheu reversão da vasectomia (n=31) ou fertilização in vitro com recuperação espermática por aspiração percutânea de espermatozoides do epidídimo (n=30) para obtenção de novos filhos, e um Grupo Controle de crianças do sexo masculino de homens férteis com vasectomia programada (n=60). A pesquisa de microdeleções do cromossomo Y foi realizada por reação em cadeia da polimerase nos pais e filhos, avaliando 20 regiões do cromossomo. Resultados O resultado não revelou microdeleções do cromossomo Y em qualquer indivíduo estudado. A incidência de microdeleções do cromossomo Y em indivíduos nascidos de pais vasectomizados que sofreram reversão de vasectomia ou fertilização in vitro com espermatozoides recuperados pela aspiração percutânea de espermatozoides do epidídimo não diferiu entre os grupos, e não houve nenhuma diferença entre indivíduos controle nascidos de gestações naturais ou incidência populacional em homens férteis. Conclusão Não foi encontrada nenhuma associação considerando microdeleções da região do fator de azoospermia no cromossomo Y e reprodução assistida. Não houve correlação entre microdeleções do cromossomo Y e vasectomia, o que sugere que as técnicas de reprodução assistida não aumentam a incidência de microdeleções do cromossomo Y.

Intracellular calcium is a target of modulation of apoptosis in MCF-7 cells in the presence of IgA adsorbed to polyethylene glycol.

PURPOSE: Clinical and epidemiological studies have indicated that breastfeeding has a protective effect on breast cancer risk. Protein-based drugs, including antibodies, are being developed to attain better forms of cancer therapy. Secretory IgA (SIgA) is the antibody class in human breast milk, and its activity can be linked to the protective effect of breastfeeding. The aim of this study was to investigate the effect of polyethylene glycol (PEG) microspheres with adsorbed SIgA on MCF-7 human breast cancer cells. METHODS: The PEG microspheres were characterized by flow cytometry and fluorescence microscopy. The MCF-7 cells were obtained from American Type Culture Collection. MCF-7 cells were pre-incubated for 24 hours with or without SIgA (100 ng/mL), PEG microspheres or SIgA adsorbed in PEG microspheres (100 ng/mL). Viability, intracellular calcium release, and apoptosis in MCF-7 cells were determined by flow cytometry. RESULTS: Fluorescence microscopy and flow cytometry analyses revealed that SIgA was able to adsorb to the PEG microspheres. The MCF-7 cells that were incubated with PEG microspheres with adsorbed SIgA showed decreased viability. MCF-7 cells that were incubated with SIgA or PEG microspheres with adsorbed SIgA had increased intracellular Ca(2+) levels. In the presence of SIgA, an increase in the percentage of apoptotic cells was observed. The highest apoptosis index was observed when the cells were treated with PEG microspheres with adsorbed SIgA. CONCLUSION: These data suggest that colostral SIgA adsorbed to PEG microspheres has antitumor effects on human MCF-7 breast cancer cells and that the presence of large amounts of this protein in secreted breast milk may provide protection against breast tumors in women who breastfed.

Incidence of Y-chromosome microdeletions in children whose fathers underwent vasectomy reversal or in vitro fertilization with epididymal sperm aspiration: a case-control study.

OBJECTIVE: To evaluate the incidence of Y-chromosome microdeletions in individuals born from vasectomized fathers who underwent vasectomy reversal or in vitro fertilization with sperm retrieval by epididymal aspiration (percutaneous epididymal sperm aspiration). METHODS: A case-control study comprising male children of couples in which the man had been previously vasectomized and chose vasectomy reversal (n=31) or in vitro fertilization with sperm retrieval by percutaneous epididymal sperm aspiration (n=30) to conceive new children, and a Control Group of male children of fertile men who had programmed vasectomies (n=60). Y-chromosome microdeletions research was performed by polymerase chain reaction on fathers and children, evaluating 20 regions of the chromosome. RESULTS: The results showed no Y-chromosome microdeletions in any of the studied subjects. The incidence of Y-chromosome microdeletions in individuals born from vasectomized fathers who underwent vasectomy reversal or in vitro fertilization with spermatozoa recovered by percutaneous epididymal sperm aspiration did not differ between the groups, and there was no difference between control subjects born from natural pregnancies or population incidence in fertile men. CONCLUSION: We found no association considering microdeletions in the azoospermia factor region of the Y chromosome and assisted reproduction. We also found no correlation between these Y-chromosome microdeletions and vasectomies, which suggests that the assisted reproduction techniques do not increase the incidence of Y-chromosome microdeletions. OBJETIVO: Avaliar a incidência de microdeleções do cromossomo Y em indivíduos nascidos de pais vasectomizados submetidos à reversão de vasectomia ou fertilização in vitro com recuperação de espermatozoides por aspiração do epidídimo (aspiração percutânea de espermatozoides do epidídimo). MÉTODOS: Estudo caso-controle que compreende crianças do sexo masculino de casais em que o homem havia sido previamente vasectomizado e escolheu reversão da vasectomia (n=31) ou fertilização in vitro com recuperação espermática por aspiração percutânea de espermatozoides do epidídimo (n=30) para obtenção de novos filhos, e um Grupo Controle de crianças do sexo masculino de homens férteis com vasectomia programada (n=60). A pesquisa de microdeleções do cromossomo Y foi realizada por reação em cadeia da polimerase nos pais e filhos, avaliando 20 regiões do cromossomo. RESULTADOS: O resultado não revelou microdeleções do cromossomo Y em qualquer indivíduo estudado. A incidência de microdeleções do cromossomo Y em indivíduos nascidos de pais vasectomizados que sofreram reversão de vasectomia ou fertilização in vitro com espermatozoides recuperados pela aspiração percutânea de espermatozoides do epidídimo não diferiu entre os grupos, e não houve nenhuma diferença entre indivíduos controle nascidos de gestações naturais ou incidência populacional em homens férteis. CONCLUSÃO: Não foi encontrada nenhuma associação considerando microdeleções da região do fator de azoospermia no cromossomo Y e reprodução assistida. Não houve correlação entre microdeleções do cromossomo Y e vasectomia, o que sugere que as técnicas de reprodução assistida não aumentam a incidência de microdeleções do cromossomo Y.

The influence of sleep restriction on expression of apoptosis regulatory proteins p53, Bcl-2 and Bax following rat tongue carcinogenesis induced by 4-nitroquinoline 1-oxide.

PURPOSE: The aim of this study was to evaluate whether sleep restriction (SR) could affect the mechanisms and pathways' essentials for cancer cells in tongue cancer induced by 4-nitroquinoline 1-oxide in Wistar rats. METHODS: The animals were distributed into 4 groups of 5 animals each treated with 50 ppm 4 NQO solution through their drinking water for 4 and 12 weeks. The animals were submitted to sleep restriction for 21 days using the modified multiple platform method, which consisted of placing 5 rats in a cage (41 × 34 × 16 cm) containing 10 circular platforms (3.5 cm in diameter) with water 1 cm below the upper surface. The investigations were conducted using immunohistochemistry of p53, Bax and Bcl-2 proteins related to apoptosis and its pathways. RESULTS: Although no histopathologic abnormalities were induced in the epithelium after 4 weeks of carcinogen exposure in all groups, in 12 weeks were observed pre-neoplastic lesions. Data analysis revealed statistically significant differences (P < 0.05) in 4 weeks group for p53, and for bcl-2. Following 12 weeks of 4NQO administration, we found significant differences between SR and control groups in p53, bax, and bcl-2 immunoexpression. CONCLUSION: Our results reveal that sleep restriction exerted alterations in proteins associated with proliferation and apoptosis in carcinogenesis.

Analysis of FokI polymorphism of vitamin D receptor gene in intervertebral disc degeneration.

AIM: We have hypothesized a possible relationship between disc degeneration (DD) and VDR FokI/T2C polymorphism. METHODS: A case-control study was performed comprising 121 Brazilian patients with confirmed DD by nuclear magnetic resonance and a control group consisting of 131 healthy patients without a history of disc cysts of the lumbar spine. Detection of VDR FokI/T2C polymorphism was performed using restriction fragment length polymorphism-polymerase chain reaction. The chi-square test was used to compare allele and genotype frequencies between groups, and a p-value of <0.05 was considered statistically significant. RESULTS: The results disclosed statistical difference between allele distribution among cases and controls (p=0.025, odds ratio=1.58, confidence interval=1.07-2.32) considering VDR FokI/T2C polymorphism. CONCLUSION: The results showed a positive association between VDR FokI/T2C polymorphism and DD in Brazilian patients tested.

Concentrated grape juice (G8000™) reduces immunoexpression of iNOS, TNF-alpha, COX-2 and DNA damage on 2,4,6-trinitrobenzene sulfonic acid-induced-colitis.

Inflammatory bowel disease (IBD) is a common chronic gastrointestinal disorder characterized by alternating periods of remission and active intestinal inflammation. Flavonoids exert several biological activities, which are mainly related to their ability to inhibit inflammatory process and/or to their antioxidant properties, and are able to regulate the immune response. The aim of this study was to evaluate whether phenolic compounds present in grape juice could reduce the inflammatory effects induced by experimental colitis. A total of 41 male Wistar rats were randomized into seven groups, as follows: G1--Sham group: sham induced-colitis rats; G2--(2,4,6-rinitrobenzenesulfonic acid) TNBS group: nontreated induced-colitis; G3--2% grape juice control group; G4--1% grape juice 24h after TNBS colitis induction; G5--1% grape juice on day 7 after colitis induction; G6--2% grape juice 24h after colitis induction; G7--2% grape juice on day 7 after colitis induction. Genotoxicity was evaluated by comet assay. Immunohistochemistry was determined using the streptavidin-biotin-peroxidase method being analyzed in control (normal tissue) and "hot spot" areas i.e., presenting inflammatory process being graded as 1 (weak), 2 (moderate), or 3 (strong). Both parameters were evaluated in the cytoplasm of epithelial or inflammatory cells. TNF-immunoexpression and iNOS were reduced after drinking grape juice 24 h or after 7 days for all doses tested. COX-2 was reduced in the groups exposed to 1% grape juice 24 h or 7 days of exposure. The grape juice at 1% dose in the last 7 days of treatment as well as grape juice at 2% dose decreased the peripheral blood genotoxicity. Taken together, the grape juice mainly at 1% dose exerts anti-inflammatory effects in chronic colitis caused by TNBS as a result of down regulation in the expression of pro-inflammatory cytokines and reduction of genotoxicity in peripheral blood cells.

Importância da Listeria monocytogenes na indústria da carne bovina / Importance of Listeria monocytogenes in the beef industry

A listeriose é uma saprozoonose de origem alimentar relativamente rara, porém grave, com elevada taxa de mortalidade. Afeta principalmente grupos de risco bem definidos, como gestantes e fetos, imunodeprimidos e idosos. Embora a maioria dos surtos esteja associada à ingestão de produtos prontos para o consumo, a Listeria sp. pode ser transmitida por uma extensa variedade de alimentos, dentre eles os de origem animal como carne, leite, queijo, com destaque para os alimentos processados. A Listeria monocytogenes é a espécie de maior importância dentro do gênero, a qual tem papel relevante na indústria da carne pela formação de biofilmes. Em bovinos de corte, a infecção alimentar não é bem descrita, no entanto, as práticas nutricionais têm sido mencionadas como as causas mais prováveis. Apesar de sua patogenicidade, no Brasil não há uma preocupação adequada com sua disseminação em abatedouros-frigoríficos, refletido pelo programa de "tolerância zero" em produtos prontos para o consumo, não aplicável às carnes in natura. Contudo, é necessária uma maior preocupação por parte das autoridades em saúde pública no país e o desenvolvimento de pesquisas que forneçam subsídios para mudanças na legislação atual.

Listeriosis is a foodborne saprozoonoses relatively rare, but serious, with a high mortality rate. It mainly affects well-defined risk groups, such as pregnant women and fetuses, older people and immunocompromised patients. Although most outbreaks are associated with ingestion of products ready for the consumption, Listeria sp. can be transmitted by a wide variety of foods, including those of animal origin such as meat, milk, cheese, especially processed foods. Listeria monocytogenes is the most important species in the genus, which has an important role in the beef industry through the formation of biofilms. Despite its pathogenicity in Brazil, there is not an adequate concern for its spread in abattoirs, reflected by the program of "zero tolerance" products ready for consumption, does not apply to meat "in natura". However, there should be more concern on the part of public health authorities in the country and the development of research grants that provide for changes in current legislation.