Occupational exposure to pesticides deregulates systemic cortisol levels in women with breast cancer and correlates with poor prognosis features

Pesticides have been pointed out as hormone disruptors and may significantly affect the prognosis of hormone-dependent diseases such as breast cancer (BC). Here, we investigated the impact of occupational pesticide exposure on systemic cortisol levels in female rural workers diagnosed with BC. Occupational exposure was assessed by interviews with a standardized questionnaire. Plasma samples (112 from pesticide-exposed women and 77 from unexposed women) were collected in the afternoon, outside the physiological cortisol peak, and analyzed by a chemiluminescent paramagnetic immunoassay for the quantitative determination of cortisol levels in serum and plasma. The results from both groups were categorized according to patients' clinicopathological and exposure data. BC pesticide-exposed women presented higher levels of cortisol than the unexposed. Higher cortisol levels were also detected in the exposed group with more aggressive disease (triple-negative BC), with tumors over 2 cm, with lymph node metastases, and with high risk of disease recurrence and death. These findings demonstrated that there is an association between pesticide exposure and BC that affected cortisol levels and correlated to poor disease prognosis.

Fish attractors when resources abound: prevalence of juveniles and lack of assemblage structure in a field experiment in the Amazon floodplain / Atratores de peixes quando os recursos são abundantes: prevalência de juvenis e falta de estrutura da assembleia em um experimento de campo na planície de inundação amazônica

Abstract Floating structures, both natural and artificial, attract fish by providing shelter, feeding areas, and nesting sites. Occupancy can be either more permanent, leading to a gradual gathering of the assembly, or transient, occurring randomly. The ability of an attractor to hold a fish assemblage may depend on the availability of habitat resources in the environment. New artificial attractors are more valuable when natural ones are scarce. Additionally, fish characteristics play a role; young and small fishes may need new habitat for shelter more than adult fish. As aquatic herbaceous areas are abundant during high water, coinciding with the period of fish reproduction, they are particularly relevant for juveniles. We constructed fish attractors composed of natural materials to investigate the structure of fish assemblages during the flood of an Amazonian floodplain lake. Our aim was to test the hypothesis that assembly processes during the flood period would be random, with a predominance of juveniles in the attractors. We collected fish at intervals of 5, 15, and 30 days, resulting in 39 observations, and classified them as either adult or juvenile. Species composition was compared among treatments using Principal Coordinates Analysis (PCoA). The assembly process was tested through species co-occurrence patterns employing null models and the C-score index. The proportion of juveniles and adults was compared using a Chi-square test. Species composition remained consistent throughout the experiment. The assembly was random, with a prevalence of juveniles, possibly serving only as temporary shelter and feeding areas. Our study contributes to understanding the role of habitat availability for floodplain fishes during high waters. The results suggest that floating attractors and new habitats may be more valuable for the juveniles than adult fish and can be used as a management strategy for population recovery, especially when floating herbaceous habitats are scarce.

Resumo As estruturas flutuantes, naturais ou artificiais, atraem os peixes, fornecendo abrigo, áreas de alimentação e locais de nidificação. A ocupação pode ser mais permanente, resultando em um recolhimento gradativo da assembleia, ou transitória, ocorrendo aleatoriamente. A capacidade de um atrator de manter uma assembleia de peixes pode depender da disponibilidade de recursos de habitat no ambiente. Novos atratores artificiais são mais valiosos quando os naturais são escassos. Além disso, as características dos peixes desempenham um papel, já que peixes jovens e pequenos podem necessitar de novos habitats como abrigo mais do que peixes adultos. Como as áreas com herbáceas aquáticas são abundantes durante as cheias, coincidindo com o período de reprodução dos peixes, elas são especialmente relevantes para peixes juvenis. Construímos atratores de peixes compostos de material natural para investigar a estrutura das assembleias de peixes durante a cheia de um lago de várzea amazônico, a fim de testar a hipótese de que durante o período de cheia, os processos de montagem seriam aleatórios e com predominância de juvenis nos atratores. Os peixes foram coletados em intervalos de 5, 15 e 30 dias, resultando em 39 observações, e classificados como adultos ou juvenis. A composição de espécies foi comparada entre os tratamentos usando uma Análise de Coordenadas Principais (PCoA). O processo de montagem foi testado por meio de padrões de coocorrência de espécies usando modelos nulos e o índice C-score. A proporção de jovens e adultos foi comparada usando um teste Qui-quadrado. A composição de espécies permaneceu a mesma ao longo do experimento. A montagem da assembleia foi aleatória com prevalência de juvenis nos atratores, que possivelmente serviam apenas como abrigo temporário e áreas de alimentação. Nosso estudo contribui para entender o papel da disponibilidade de novos habitats para peixes de várzea durante a cheia. Os resultados sugerem que atratores flutuantes e novos habitats podem ser mais valiosos para os peixes jovens do que para adultos e podem ser usados como estratégia de manejo para a recuperação populacional, especialmente quando habitats de herbáceas flutuantes são escassos.

First molecular-based detection study of Leishmania infantum in the Tapirapé indigenous population in the Brazilian Amazon

Species of the genus Leishmania parasitize mammals and have life cycles that alternate between vertebrate and invertebrate hosts. Most species develop in a hematophagous arthropod and infect a specific vertebrate host that may belong to diverse orders and families. Visceral leishmaniasis is a chronic zoonosis with a wide geographic distribution, affecting 350 million people globally, mostly in areas with a high risk of infection. In Brazil, this disease not only has a high incidence but is also expanding to new areas, both in urban centers and rural areas, including territories with tribal communities, due to increasing human intervention. The objective of this study was to perform cathepsin L-like gene-based molecular diagnosis of Leishmania infantum in the indigenous Tapirapé ethnic group in the state of Mato Grosso. From the 372 individuals assessed, only 0.8% (3/372) tested positive for L. infantum, all from the same village (Urubu Branco). Despite the small number of infected individuals, this study demonstrates the first human cases of Leishmania infantum infection in this population, suggesting the need for regular monitoring of visceral leishmaniasis in the area and leading to a broad discussion on the planning and implementation of public health measures for the indigenous population, while respecting their distinctive territories and culture.

Persistent symptoms and decreased health-related quality of life after symptomatic pediatric COVID-19: A prospective study in a Latin American tertiary hospital

OBJECTIVES: To prospectively evaluate demographic, anthropometric and health-related quality of life (HRQoL) in pediatric patients with laboratory-confirmed coronavirus disease 2019 (COVID-19) METHODS: This was a longitudinal observational study of surviving pediatric post-COVID-19 patients (n=53) and pediatric subjects without laboratory-confirmed COVID-19 included as controls (n=52) was performed. RESULTS: The median duration between COVID-19 diagnosis (n=53) and follow-up was 4.4 months (0.8-10.7). Twenty-three of 53 (43%) patients reported at least one persistent symptom at the longitudinal follow-up visit and 12/53 (23%) had long COVID-19, with at least one symptom lasting for >12 weeks. The most frequently reported symptoms at the longitudinal follow-up visit were headache (19%), severe recurrent headache (9%), tiredness (9%), dyspnea (8%), and concentration difficulty (4%). At the longitudinal follow-up visit, the frequencies of anemia (11% versus 0%, p=0.030), lymphopenia (42% versus 18%, p=0.020), C-reactive protein level of >30 mg/L (35% versus 0%, p=0.0001), and D-dimer level of >1000 ng/mL (43% versus 6%, p=0.0004) significantly reduced compared with baseline values. Chest X-ray abnormalities (11% versus 2%, p=0.178) and cardiac alterations on echocardiogram (33% versus 22%, p=0.462) were similar at both visits. Comparison of characteristic data between patients with COVID-19 at the longitudinal follow-up visit and controls showed similar age (p=0.962), proportion of male sex (p=0.907), ethnicity (p=0.566), family minimum monthly wage (p=0.664), body mass index (p=0.601), and pediatric pre-existing chronic conditions (p=1.000). The Pediatric Quality of Live Inventory 4.0 scores, median physical score (69 [0-100] versus 81 [34-100], p=0.012), and school score (60 [15-100] versus 70 [15-95], p=0.028) were significantly lower in pediatric patients with COVID-19 at the longitudinal follow-up visit than in controls. CONCLUSIONS: Pediatric patients with COVID-19 showed a longitudinal impact on HRQoL parameters, particularly in physical/school domains, reinforcing the need for a prospective multidisciplinary approach for these patients. These data highlight the importance of closer monitoring of children and adolescents by the clinical team after COVID-19.

Experimental infection by Anaplasma marginale in buffaloes and cattle: clinical, hematological, molecular and pathological aspects

ABSTRACT: The study aimed to evaluate and compare the clinical, laboratory and pathological aspects of buffalo and bovine experimentally infected with AmRio 2 strain of Anaplasma marginale. Four Murrah buffaloes and four crossbred cattle were used in the experiment, which two animals of each species were splenectomized. Strain AmRio 2 of A. marginale was inoculated in all experimental animals. Clinical exams, Packed Cell Volume (PCV), blood counts, blood smears, rickettsemia, necropsy and histopathology were performed in all cases. Semi-Nested-PCR (snPCR) for the msp5 and snPCR for the msp1 target gene for identification of A. marginale in blood samples from animals was done. From positive samples for msp1 snPCR, samples were analyzed for the amino acid sequences of this gene. Two splenectomized cattle presented apathy, pale mucous membranes, jaundice, hyperthermia, and severe anemia. The remaining experimental animals did not show clinical signs. The rickettsemia in all animals was less than 1%. The mean PCV of the splenectomized cattle was below 20% at two-time points after infection. On the blood count, the main changes were observed in splenectomized calves and were characterized by a decrease in red blood cells, hemoglobin, PCV and platelets (p 0.05). All animals presented leukocyte elevation by increased lymphocytes, however, with no significant difference. The average prepatent period was two days in all the animals. The average incubation period in cattle that became ill was 25.5 days, and death occurred, on average, 63 days after inoculation of the strain. The necropsy findings were characterized by pale carcass, ascites, enlarged liver, distended gallbladder, and thick bile. Histopathological findings included infiltration of macrophages and lymphocytes in various organs, hepatic sinusoidal dilatation, and necrosis of the large intestine. In snPCR for the msp5 gene, 100% of the animals were positive in at least one evaluation. And in the snPCR for the infection of the msp1 target gene was also found in all animals in at least one sample evaluated. However, sequencing revealed only five animals, including the bovine which died, with a similarity of the amino acid sequences with AmRio 2 strain of A. marginale. It is concluded that the splenectomized cattle died due to anaplasmosis caused by the inoculated strain and the buffalo were more resistant compared to cattle. Buffaloes can be an alternative to cattle rearing in areas with a high occurrence of clinical cases of anaplasmosis.

RESUMO: O estudo teve como objetivo avaliar e comparar os aspectos clínicos, laboratoriais e patológicos de búfalos e bovinos infectados experimentalmente com estirpe AmRio 2 de Anaplasma marginale. Para isso, foram utilizados quatro bubalinos Murrah e quatro bovinos mestiços, sendo dois animais de cada espécie, esplenectomizados. Estirpe AmRio 2 de A. marginale foi inoculada em todos os animais. Foram realizados exames clínicos, hematócrito, hemograma, esfregaço sanguíneo com avaliação de riquetsemia, necropsia e histopatologia, além de, Semi-Nested-PCR (snPCR) para o gene alvo msp5 e snPCR para o gene alvo msp1 para identificação de A. marginale nas amostras de sangue dos ruminantes. A partir das amostras positivas na snPCR msp1, foram selecionadas amostras para análise das sequências de aminoácidos deste gene. Dois bovinos esplenectomizados apresentaram apatia, mucosas pálidas, icterícia, hipertermia e anemia severa. O restante dos animais não apresentou sintomatologia clínica. A riquetsemia em todos os animais foi menor que 1%. A média do hematócrito dos bovinos esplenectomizados esteve abaixo de 20% em dois momentos após infecção. Ao hemograma, as principais alterações observadas foram nos bovinos esplenectomizados e caracterizaram-se por redução de hemácias, hemoglobina, hematócrito e plaquetas (p 0,05). Todos os animais apresentaram elevação de leucócitos por aumento de linfócitos, porém, sem diferença significativa. O período pré-patente médio foi de dois dias em todos os animais. O período de incubação médio nos bovinos que adoeceram foi de 25,5 dias e estes morreram em média 63 dias após inoculação da estirpe. Os achados de necropsia caracterizaram-se por carcaça pálida, ascite, aumento de volume do fígado, vesícula biliar distendida e bile espessa. À histopatologia, verificou-se infiltração de macrófagos e linfócitos em diversos órgãos, dilatação dos sinusoides hepáticos e necrose do intestino grosso. A snPCR para o gene msp5, revelou 100% dos animais positivos em pelo menos um momento de avaliação. E na snPCR para o gene alvo msp1 também verificou-se infecção em todos os animais em pelo menos uma amostra avaliada. Entretanto, o sequenciamento revelou apenas cinco animais, incluindo os bovinos que morreram, com similaridade das sequências de aminoácidos com estirpe AmRio 2 de A. marginale. Conclui-se que os bovinos esplenectomizados morreram em virtude de anaplasmose provocada pela estirpe inoculada e os bubalinos foram mais resistentes em comparação aos bovinos. Finalmente, os búfalos podem ser uma alternativa à criação de bovinos em áreas com alta ocorrência de casos clínicos de anaplasmose.

Bócio em bovinos

ABSTRACT: Goiter is a non-inflammatory and non-neoplastic enlargement of the thyroid gland in adults and neonates. One of the main causes involved is the nutritional deficiency of iodine. In this study three outbreaks of goiter in cattle are reported. In the first outbreak (Farm A) 60 calves were affected, of which 20 died soon after birth, 30 recovered and 10 remained sick. In the second outbreak (Farm B) a cow and its fetus were affected and in the third outbreak (Farm C) two calves were affected. The main clinical signs observed were bilaterally enlarged thyroid glands, weight loss, respiratory distress, hypotrichosis and myxedema. Additionally in the Farm A abortions were reported. Macroscopically the thyroid of the calves and fetus was enlarged, dark red with evident vascularization and cervical subcutaneous edema. In the Farm B the cow showed enlarged thyroid gland as well. Microscopically the calves and the fetus had hyperplastic and heterogeneous thyroid follicles with absence of colloid and vascularized interstitium. The thyroid of the cow was also hyperplastic, but had higher amount of colloid. In all outbreaks the mineral supplementation was performed by mixing non-iodized white salt with the mineral mixture. On the property A the white salt was non-iodized and mixed in equal parts with the mineral salt. At property B the salt was also mixed to a non-iodized white salt in ratio and 1:2, respectively. In the property C, mineral salt and non-iodized white salt were offered in separate troughs in the field, and according to the owners report the animals prioritized the consumption of white salt. In all the monitored properties it was recommended to stop the addition of white salt in the mineral mixture and the administration of iodine to the affected calves. After these measurements no new cases were observed in the properties. Despite being a well-known disease, there are still few reported cases of goiter in cattle in Brazil. Moreover, little is known about the actual mineral deficiencies of each region, and that simple nutritional management guidelines are still needed.

RESUMO: Bócio é o aumento não inflamatório e não neoplásico da glândula tireoide em animais adultos e recém-nascidos. Uma das principais causas envolvidas é a deficiência nutricional de iodo. Relata-se neste trabalho, a ocorrência de três surtos de bócio em bovinos. Na primeira propriedade (Propriedade A), 60 bezerros foram afetados, sendo que 20 morreram logo após o nascimento, 30 recuperaram-se e 10 permaneceram doentes e tiveram remissão dos sinais apenas após tratamento parenteral com iodo. Na segunda propriedade (Propriedade B) uma vaca e seu feto foram acometidos e na terceira (Propriedade C) dois bezerros foram afetados. Os principais sinais clínicos observados nos bezerros foram aumento de volume bilateral na região cervical ventral, emagrecimento, dificuldade respiratória, hipotricose e desenvolvimento corpóreo retardado. Na propriedade A foi relatada ainda a ocorrência de abortamentos. Macroscopicamente, os bezerros e o feto apresentavam tireoide aumentada, vermelho-escura, com vascularização evidente e edema subcutâneo cervical. No surto dois também foi observado, à necropsia, aumento de volume da glândula tireoide da vaca. Microscopicamente, a tiroide dos bezerros e do feto apresentava folículos tireoidianos hiperplásicos, heterogêneos, destituídos de coloide e com interstício acentuadamente vascularizado. A tireoide da vaca era semelhante à dos bezerros, no entanto, possuía quantidade maior de coloide. Em todos os casos a suplementação mineral era realizada por meio da mistura de sal mineral com sal branco. Na Propriedade A o sal branco era não iodado e misturado em partes iguais com o sal mineral. Na Propriedade B o sal também era misturado a um sal branco não iodado na proporção e 1:2, respectivamente. Na Propriedade C o sal mineral e o sal branco não iodado eram ofertados em cochos separados no campo e, segundo relato do proprietário, os animais priorizavam o consumo do sal branco. Em todas as propriedades acompanhadas foi recomendada a interrupção da adição de sal branco na mistura mineral e a administração de iodo aos bezerros acometidos. Após essas medidas não foram observados novos casos nas propriedades. Apesar de ser uma enfermidade bem conhecida, ainda são poucos os relatados de casos de bócios em bovinos no Brasil. Além disso, pouco se sabe sobre as reais deficiências minerais de cada região, e que simples orientações de manejo nutricional ainda são necessárias.

Could formaldehyde induce mutagenic and cytotoxic effects in buccal epithelial cells during anatomy classes?

BACKGROUND: Due to increased formaldehyde exposure, carcinogenic to humans, several researches have been studying the potential toxicity and the safe levels for human beings. The aim of this study was to investigate mutagenicity and cytotoxicity in buccal epithelial exfoliated cells (BEC) of students subjected to formaldehyde (FA) during anatomy classes. MATERIAL AND METHODS: BEC were collected periodically from 17 volunteers of undergraduate programs, who had participated in practical anatomy classes, before and after FA exposure. Cells were stained according to Feulgen method and then micronucleus test was applied. A total of 1,500 cells were assessed per individual in this study for the micronucleus frequency and other parameters of cytotoxicity. RESULTS: There was statistically significant increase in number of micronucleated BEC after FA exposure (after 1 month p=.034 and after 3.5 months p=.017). However, FA exposure caused no significant increase in other nuclear alterations closely related to cytotoxicity (p≥.05). CONCLUSIONS: FA induced mutagenicity during anatomy classes. Cell death increased, but it was not statistically significant. Efforts have to be made to improve air quality and reduce exposures during anatomy classes.

Morfologia de órgãos genitais femininos de quelônio semi-aquático Mesoclemmys vanderhaegei

ABSTRACT: Mesoclemmys vanderhaegei (Testudines: Chelidae) is a freshwater turtle with occurrence in Amazon, Tocantins, Paraguai, Paraná and Uruguai rivers basins. Although according to International Union for Conservation of Nature, it has low risk of extinction, there is an uptade necessity of ecological and biological data. Considering that the management and conservation plans are related to a wide knowledge of reproductive biology, a first macroscopic description about the young and adults females of M. vanderhaegei is important. These points were correlated to the specimens size and period of the year. The samples of M. vanderhaegei were collected in Chapada dos Guimarães county, Mato Grosso, Brazil, an area of large natural occurrence of the specimens. Genital organs of seventeen females were fixed in 10% formalin and then dissected to demonstrate the particularities related to external and internal anatomy. The young and adult M. vanderhaegei genitals organs are composed of ovaries and oviducts pairs that dorsolaterally discharge into the cloaca, forming with the ureter, the urogenital papilla. The ovaries are elongated organs with larger cranial and elongated caudal portions. The oviducts, which are in adults very differentiated in its shape and size compared to the young, are long and located laterally to the ovaries. In both age groups, the genital organs are supported by celomatic membrane folds that emerge from the ceiling of the cavity, constituting the mesovary and mesoviduct. In adult females, according to the shape and pattern of the oviduct mucosa, the cranial segment corresponds to the regions of the infundibulum and magnum; the middle segment, the isthmus and the caudal segment identify the uterus and vagina regions. The clitoris is sited on the floor of the urodeum. The carapace linear length and body mass between immature and adult females vary to 134-155.6mm e 134.43-365g respectively. The main part of young females was captured in rainy period and the adults, with and without eggs, at dry period. The macroscopic characteristics of the M. vanderhaegei genital organs are also observed in others Testudines, with the exception of urogenital papilla and by the clitoris presence.

RESUMO: O Mesoclemmys vanderhaegei (Testudines: Chelidae) é um quelônio de água doce cuja ocorrência é descrita nas bacias dos rios Amazonas, Tocantins, Paraguai, Paraná e Uruguai. Consta na lista vermelha de espécies ameaçadas da International Union for Conservation of Nature, como espécie de baixo risco, mas que poderá se tornar ameaçada, sendo necessário a atualização de seus dados ecológicos e biológicos. Considerando que planos de manejo e conservação de espécies dependem também de vasto conhecimento sobre a biologia reprodutiva, apresentamos a primeira descrição macroscópica dos órgãos genitais de fêmeas jovens e adultas de M. vanderhaegei correlacionando esses achados ao tamanho do espécime e ao período do ano. Exemplares de M. vanderhaegei foram coletados no município de Chapada dos Guimarães, área de ampla ocorrência natural da espécie. A descrição dos órgãos genitais foi realizada a partir de 17 fêmeas fixadas em formol a 10% e dissecadas para evidenciação de particularidades relacionadas à anatomia externa e interna. Os órgãos genitais M. vanderhaegei jovens e adultas são constituídos por pares de ovários e ovidutos que desembocam dorsolateralmente na cloaca, formando junto com o ureter a papila urogenital. Os ovários são órgãos alongados, com a extremidade cranial mais larga e caudal afunilada, já os ovidutos são longos e localizados lateralmente aos ovários, sendo em adultas, muito diferenciado na sua forma e tamanho em relação ao das jovens. Em ambas faixas etárias, os órgãos genitais são sustentados por pregas de membrana celomática que emergem do teto da cavidade, constituindo os mesovário e mesoviduto. De acordo com a forma e o padrão de mucosa do oviduto em fêmeas adultas, o segmento cranial corresponde as regiões do infundíbulo e magnum, o segmento médio, ao istmo e no caudal identificam-se as regiões útero e vagina. No assoalho do urodeum aloja-se o clitóris. O comprimento retilíneo de carapaça e massa corpórea entre fêmeas imaturas e adultas variam, respectivamente, entre 134-155,6mm e 134,43-365g. A maioria das fêmeas jovens foram capturadas no período chuvoso e as adultas sem e com ovos no período seco. As características macroscópicas dos órgãos genitais observadas em M. vanderhaegei são compartilhadas com outros Testudines, exceto pela papila urogenital e presença do clitóris.

Building the SeqChromMM Markov property atlas of the human genome by analyzing the 200-bp units of the 15 different chromatin regions of ENCODE.

We analyzed the publicly available ChromHMM BED files of the ENCODE project and tested the Markov properties of the different chromatin states in the human genome. Nucleotide frequency profiles of regional chromatin segmentations were analyzed, and Markov chains were built to detect Markov properties in the chromatin states of different ChromHMM regions. By estimating the transition probabilities of 200-base pair nucleotide sequences of the human genome, we constructed a nucleotide-sequence-based Markovian chromatin map called SeqChromMM.

Using Markov chains of nucleotide sequences as a possible precursor to predict functional roles of human genome: a case study on inactive chromatin regions.

Recent advances in computational epigenetics have provided new opportunities to evaluate n-gram probabilistic language models. In this paper, we describe a systematic genome-wide approach for predicting functional roles in inactive chromatin regions by using a sequence-based Markovian chromatin map of the human genome. We demonstrate that Markov chains of sequences can be used as a precursor to predict functional roles in heterochromatin regions and provide an example comparing two publicly available chromatin annotations of large-scale epigenomics projects: ENCODE project consortium and Roadmap Epigenomics consortium.

Clinical characteristics of men with non-mosaic Klinefelter syndrome in northeastern China: implications for genetic counseling.

Klinefelter syndrome (KS) is the most common genetic cause of male infertility. Widespread development in assisted reproductive technology has provided non-mosaic KS patients with the opportunity of having biological children. Testosterone replacement therapy and micro-dissection testicular sperm extraction are effective sperm retrieval techniques for KS patients. Despite the success of sperm retrieval and intracytoplasmic sperm injection (ICSI), some areas of early aggressive hormonal spermatogenesis and appropriate management of KS remain controversial. Androgenotherapy, a common treatment for KS, carries a risk of decreasing focal spermatogenesis by lowering the gonadotropin content. Inadequately treated hypogonadism increases psychosocial morbidity in KS patients. Preventive care must be provided from the time of diagnosis, preferentially through a multidisciplinary approach. This indicates the need for improved genetic counseling of KS patients. The aim of this study was to report the prevalence of non-mosaic KS in a Chinese infertile male population. The rate of early diagnosis was lower in KS patients; most of these were diagnosed after rising concerns of reproductive capacity. The mean age of patients with sperm or germ cells was significantly lower, while the semen volume of these patients was significantly higher. However, the semen volume was negatively correlated with the age and ratio of luteinizing hormone/testosterone content in KS patients. Therefore, genetic counseling of KS patients should focus on early diagnosis and timely treatment, in addition to improving the quality of life of all KS patients. The use of testosterone replacement therapy and/ or micro-dissection testicular sperm extraction should be preferentially considered for fertility preservation.

Transplantation of umbilical cord blood mononuclear cells increases levels of nerve growth factor in the cerebrospinal fluid of patients with autism.

We aimed to evaluate the levels of growth factors in the cerebrospinal fluid (CSF) of patients with autism after transplantation of umbilical cord blood mononuclear cells (CBMNCs). Fourteen subjects diagnosed with autism received transplantation of CBMNCs first through intravenous infusion, and three times subsequently through intrathecal injections. A 2-mL sample of CSF was taken before each intrathecal injection. CSF levels of nerve growth factor (NGF), vascular endothelial growth factor (VEGF), and basic fibroblast growth factor (bFGF) were determined by enzyme-linked immunosorbent assay. All data are reported as means ± SD and were analyzed using the SPSS 10.0 software. One-way analysis of variance with post-hoc F-and Q-tests were performed for comparisons. NGF levels in the CSF were significantly increased after transplantation (213.54 ± 56.38 after the third versus 28.32 ± 12.22 ng/L after the first transplantation; P < 0.05), while VEGF and bFGF levels did not change significantly. Therefore, transplantation of CBMNCs could increase NGF levels in the CSF of patients with autism.

Establishment of a rabbit model of coronary artery bypass graft and endothelial nitric oxide synthase gene transfection.

This study established an animal model of coronary artery bypass graft (CABG) surgery. The human endothelial nitric oxide synthase (eNOS) gene was transfected into grafted arterial walls to verify transfection efficiency. Forty rabbits were randomized into the following 4 equal groups: 1) eNOS gene transfection group (eNOS group); 2) empty eNOS gene transfection group (empty gene group); 3) control group; 4) normal femoral artery group. Grafted arteries, and normal carotid and femoral artery specimens were obtained 3 weeks later. Immunohistochemistry and analyses of tissue nitric oxide (NO) levels, eNOS activity, and eNOS protein western blotting were performed. The effectiveness and efficiency of transfection were observed and confirmed. All rabbits survived. The grafted arteries retained patency. Varying degrees of adaptability changes were observed in grafted arteries in each group. The eNOS group exhibited vascular wall thickening and significantly increased eNOS protein expression. The control and empty gene groups exhibited vessel wall degeneration, and eNOS protein was weakly or not expressed (P < 0.05). The arterial wall NO concentration and total eNOS activity in the eNOS group were significantly higher than those in the other groups (P < 0.05). Western blotting demonstrated that the vascular wall eNOS protein concentration was significantly greater than that in the other groups (P < 0.05). Furthermore, the eNOS gene transfection can increase eNOS expression and activity in vessel walls, increasing local NO concentration and expression.

Role of chemokines in promoting instability of coronary atherosclerotic plaques and the underlying molecular mechanism

Our aim was to investigate the role of chemokines in promoting instability of coronary atherosclerotic plaques and the underlying molecular mechanism. Coronary angiography and intravascular ultrasound (IVUS) were performed in 60 stable angina pectoris (SAP) patients and 60 unstable angina pectoris (UAP) patients. The chemotactic activity of monocytes in the 2 groups of patients was examined in Transwell chambers. High-sensitivity C-reactive protein (hs-CRP), monocyte chemoattractant protein-1 (MCP-1), regulated on activation in normal T-cell expressed and secreted (RANTES), and fractalkine in serum were examined with ELISA kits, and expression of MCP-1, RANTES, and fractalkine mRNA was examined with real-time PCR. In the SAP group, 92 plaques were detected with IVUS. In the UAP group, 96 plaques were detected with IVUS. The plaques in the UAP group were mainly lipid 51.04% (49/96) and the plaques in the SAP group were mainly fibrous 52.17% (48/92). Compared with the SAP group, the plaque burden and vascular remodeling index in the UAP group were significantly greater than in the SAP group (P<0.01). Chemotactic activity and the number of mobile monocytes in the UAP group were significantly greater than in the SAP group (P<0.01). Concentrations of hs-CRP, MCP-1, RANTES, and fractalkine in the serum of the UAP group were significantly higher than in the serum of the SAP group (P<0.05 or P<0.01), and expression of MCP-1, RANTES, and fractalkine mRNA was significantly higher than in the SAP group (P<0.05). MCP-1, RANTES, and fractalkine probably promote instability of coronary atherosclerotic plaque.

Hepatic inducible nitric oxide synthase expression increases upon exposure to hypergravity

Stimulation by a number of conditions, including infection, cytokines, mechanical injury, and hypoxia, can upregulate inducible nitric oxide synthase (iNOS) in hepatocytes. We observed that exposure to hypergravity significantly upregulated the transcription of the hepatic iNOS gene. The aim of this study was to confirm our preliminary data, and to further investigate the distribution of the iNOS protein in the livers of mice exposed to hypergravity. ICR mice were exposed to +3 Gz for 1 h. We investigated the time course of change in the iNOS expression. Hepatic iNOS mRNA expression progressively increased in centrifuged mice from 0 to 12 h, and then decreased rapidly by 18 h. iNOS mRNA levels in the livers of centrifuged mice was significantly higher at 3, 6, and 12 h than in uncentrifuged control mice. The pattern of iNOS protein expression paralleled that of the mRNA expression. At 0 and 1 h, weak cytoplasmic iNOS immunoreactivity was found in some hepatocytes surrounding terminal hepatic venules. It was noted that at 6 h there was an increase in the number of perivenular hepatocytes with moderate to strong cytoplasmic immunoreactivity. The number of iNOS-positive hepatocytes was maximally increased at 12 h. The majority of positively stained cells showed a strong intensity of iNOS expression. The expression levels of iNOS mRNA and protein were significantly increased in the livers of mice exposed to hypergravity. These results suggest that exposure to hypergravity significantly upregulates iNOS at both transcriptional and translational levels.

Effect of physical training on liver expression of activin A and follistatin in a nonalcoholic fatty liver disease model in rats

Nonalcoholic fatty liver disease (NAFLD) is characterized by fat accumulation in the liver and is associated with obesity and insulin resistance. Activin A is a member of the transforming growth factor beta (TGF)-β superfamily and inhibits hepatocyte growth. Follistatin antagonizes the biological actions of activin. Exercise is an important therapeutic strategy to reduce the metabolic effects of obesity. We evaluated the pattern of activin A and follistatin liver expression in obese rats subjected to swimming exercise. Control rats (C) and high-fat (HF) diet-fed rats were randomly assigned to a swimming training group (C-Swim and HF-Swim) or a sedentary group (C-Sed and HF-Sed). Activin βA subunit mRNA expression was significantly higher in HF-Swim than in HF-Sed rats. Follistatin mRNA expression was significantly lower in C-Swim and HF-Swim than in either C-Sed or HF-Sed animals. There was no evidence of steatosis or inflammation in C rats. In contrast, in HF animals the severity of steatosis ranged from grade 1 to grade 3. The extent of liver parenchyma damage was less in HF-Swim animals, with the severity of steatosis ranging from grade 0 to grade 1. These data showed that exercise may reduce the deleterious effects of a high-fat diet on the liver, suggesting that the local expression of activin-follistatin may be involved.

Effects and mechanisms of Fenofibrate on the secretion of vascular endothelial contraction factors in hypertensive rats.

This study investigated the effects of the peroxisome proliferator-activated receptor alpha (PPAR-α) agonist, Fenofibrate, on the secretion of vascular endothelial contraction factors in hypertensive rats to elucidate its possible mechanisms. The vascular ring contraction experiment was used to observe whether rat vascular tension of clean grade spontaneously hypertensive rats (SHR) changes after 1-h incubation of 0.1, 1.0, 10.0 µM Fenofibrate with 10.0 µM Fenofibrate, a PPAR-α antagonist (MK866), and a PPAR-γ antagonist (GW9662) in SHR. The results were compared with Wistar Kyoto rats. Enzyme-linked immunosorbent assay was used to detect the secretion of the serum vascular endothelial contraction factor prostacyclin-1α (PGF-1α), PGF-2α, and thromboxane B2 (TXB2). Western blot was used to detect COX-1 protein expression. A quantity of 10.0 µM Fenofibrate significantly reduced vasoconstriction in SHR compared to the control group (P = 0.013). The PPAR-α antagonist, MK866, significantly improved the vascular contractility of SHR when incubated with 10.0 µM Fenofibrate (P = 0.021). The PPAR-γ antagonist, GW9662, had no significant effect on the vascular contractility of SHR when incubated with 10.0 µM Fenofibrate (P = 0.071). The isolated aorta of SHR released significantly lower PGF- 1α (P = 0.014), PGF-2α (P = 0.023), and TXB2 (P = 0.017) levels in the 10.0 µM Fenofibrate group compared to the control group. COX-1 expression of SHR rat vascular endothelium was significantly depressed in the 10.0 µM Fenofibrate group compared to the control group (P = 0.027). In conclusion, Fenofibrate reduces the secretion of vascular endothelial contraction factors in hypertensive rats, which might arise through the endothelium influencing COX-1 expression.

Changes in cardiac aldosterone and its synthase in rats with chronic heart failure: an intervention study of long-term treatment with recombinant human brain natriuretic peptide

The physiological mechanisms involved in isoproterenol (ISO)-induced chronic heart failure (CHF) are not fully understood. In this study, we investigated local changes in cardiac aldosterone and its synthase in rats with ISO-induced CHF, and evaluated the effects of treatment with recombinant human brain natriuretic peptide (rhBNP). Sprague-Dawley rats were divided into 4 different groups. Fifty rats received subcutaneous ISO injections to induce CHF and the control group (n=10) received equal volumes of saline. After establishing the rat model, 9 CHF rats received no further treatment, rats in the low-dose group (n=8) received 22.5 μg/kg rhBNP and those in the high-dose group (n=8) received 45 μg/kg rhBNP daily for 1 month. Cardiac function was assessed by echocardiographic and hemodynamic analysis. Collagen volume fraction (CVF) was determined. Plasma and myocardial aldosterone concentrations were determined using radioimmunoassay. Myocardial aldosterone synthase (CYP11B2) was detected by quantitative real-time PCR. Cardiac function was significantly lower in the CHF group than in the control group (P<0.01), whereas CVF, plasma and myocardial aldosterone, and CYP11B2 transcription were significantly higher than in the control group (P<0.05). Low and high doses of rhBNP significantly improved hemodynamics (P<0.01) and cardiac function (P<0.05) and reduced CVF, plasma and myocardial aldosterone, and CYP11B2 transcription (P<0.05). There were no significant differences between the rhBNP dose groups (P>0.05). Elevated cardiac aldosterone and upregulation of aldosterone synthase expression were detected in rats with ISO-induced CHF. Administration of rhBNP improved hemodynamics and ventricular remodeling and reduced myocardial fibrosis, possibly by downregulating CYP11B2 transcription and reducing myocardial aldosterone synthesis.

A novel multiplex PCR for the simultaneous detection of Salmonella enterica and Shigella species

Salmonella enterica and Shigella species are commonly associated with food and water borne infections leading to gastrointestinal diseases. The present work was undertaken to develop a sensitive and reliable PCR based detection system for simultaneous detection of Salmonella enterica and Shigella at species level. For this the conserved regions of specific genes namely ipaH1, ipaH, wbgZ, wzy and invA were targeted for detection of Shigella genus, S. flexneri, S. sonnei, S. boydii and Salmonella enterica respectively along with an internal amplification control (IAC). The results showed that twenty Salmonella and eleven Shigella spp., were accurately identified by the assay without showing non-specificity against closely related other Enterobacteriaceae organisms and also against other pathogens. Further evaluation of multiplex PCR was undertaken on 50 natural samples of chicken, eggs and poultry litter and results compared with conventional culture isolation and identification procedure. The multiplex PCR identified the presence of Salmonella and Shigella strains with a short pre-enrichment step of 5 h in peptone water and the same samples were processed by conventional procedures for comparison. Therefore, this reported multiplex PCR can serve as an alternative to the tedious time-consuming procedure of culture and identification in food safety laboratories.