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1.
Rev. bras. plantas med ; 18(2): 433-441, 2016. graf
Article in Portuguese | LILACS | ID: lil-787945

ABSTRACT

RESUMO Considerando os diferentes usos etnofarmacológicos apresentados pela planta Luehea divaricata, realizou-se este estudo com o objetivo de avaliar as atividades antinociceptiva e antinflamatória do extrato etanólico de suas folhas, em modelo animal, nas dosagens de 20, 40, 80 e 160 mg/Kg, por via oral. Foram realizados os seguintes testes: contorções abdominais induzidas pelo acido acético, placa quente, formalina e edema de pata induzido por carragenina. Foram utilizados camundongos Swiss (20-25 g) para os três primeiros testes e ratos Wistar (180-250 g), para o último, divididos em seis grupos de oito animais, totalizando 48 animais em cada parâmetro de avaliação. Os resultados foram analisados estatisticamente pela análise de variância a 5% de probabilidade, para verificar quais os tratamentos que diferiram entre si, e estes foram submetidos aos testes de Kruskall-Wallis e Student-Newman-Keuls. O extrato etanólico das folhas de L. divaricata (EEtOH-Ld), nas diferentes doses estudadas, apresentou significativa atividade antinociceptiva sobre a dor induzida quimicamente por injeções intraperitoneal de acido acético e intraplantar de formalina. Na dosagem de 160 mg/Kg, esse extrato apresentou ação analgésica central, aos 120 minutos de observação, no teste de placa quente e reduziu o edema de pata induzido pela administração de carragenina, uma hora após a administração do agente inflamatório, semelhante ao efeito produzido pelo fármaco padrão.


ABSTRACT Considering the different ethnopharmacological uses submitted by the plant Luehea divaricata, this study took place in order to evaluate the antinociceptive and anti-inflammatory activities of the ethanol extract of the leaves in an animal model, the dosages of 20, 40, 80 and 160 mg/kg by oral intake. The following tests were performed: writhing induced by acetic acid, hot plate, formalin, and paw edema induced by carrageenan. Swiss mice (20-25 g) were used for the first three tests and Wistar rats (180-250 g) for the last, divided into six groups, each of eight animals, totaling 48 animals for each assessment parameter. The results were statistically analyzed by analysis of variance at 5% probability to verify which treatments differ, and these were tested by Kruskal-Wallis and Student-Newman-Keuls. The ethanol extract of L. divaricataleaves (EEtOH-Ld) at the different studied doses showed significant antinociceptive activity on chemically induced pain by intraperitoneal injections of acetic acid and intraplantar formalin. At a dosage of 160 mg/kg, this extract showed a central analgesic action after 120 minutes of observation in the hot plate test and reduced action in the paw edema induced by carrageenan one hour after the administration of the inflammatory agent, similar to the effect produced by the standard drug.


Subject(s)
Mice , Malvaceae/classification , Analgesics/pharmacology , Anti-Inflammatory Agents/pharmacology , Pain/pathology , Plants, Medicinal/classification
2.
Rev. bras. plantas med ; 17(3): 413-419, Jul-Sep/2015. tab
Article in Portuguese | LILACS | ID: lil-752550

ABSTRACT

RESUMO Este estudo teve como objetivo avaliar os efeitos da própolis sobre os perfis leucocitário e protéico de camundongos e sobre o tempo de fechamento de lesões de pele confeccionadas experimentalmente, limpas e infectadas com Staphylococcus aureus. No primeiro, foram utilizados 48 animais divididos em quatro grupos, sendo um tratado com solução hidroalcóolica pura e três tratados com própolis a 10%, nas dosagens de 20 mg, 40 mg e 80 mg por animal de 25 gramas de peso, em única aplicação intraperitoneal. Foram coletadas amostras de sangue no segundo, 10º, 18º e 26º dias após o tratamento para realização de leucograma, proteinograma e fracionamento eletroforético das proteínas. Na avaliação da atividade cicatrizante, também foram utilizados 48 camundongos divididos em seis grupos, nos quais realizou-se a confecção cirúrgica de feridas na dimensão de 1cm2, após anestesia dissociativa. Dois grupos serviram como controle para feridas limpas e infectadas. Dois grupos de feridas infectadas por S. aureuse dois grupos de feridas limpas foram tratados com própolis a 5% e 10%, sendo a escolha destas concentrações baseada em um estudo piloto realizado. Os resultados mostraram que o tratamento com própolis influencia o leucograma e o proteinograma, de forma dose-dependente, sendo que a maior dose utilizada desencadeou leucocitose com linfocitose e aumento de proteínas da fração gamaglobulínica, no 10º dia após o início do tratamento. Também mostraram que a concentração da solução influenciou o tempo de cicatrização das feridas infectadas, ocorrendo em menor tempo no grupo tratado com a solução a 5%.


ABSTRACT This study aimed to evaluate the propolis effects on both the leukocyte and protein profiles of mice and on the closing time of skin lesions made experimentally, clean and infected with Staphylococcus aureus. The first 48 animals were divided into four groups, one treated with pure alcohol solution and three treated with propolis 10% at dosages of 20 mg, 40 mg and 80 mg per 25 g of animal weight in an intraperitoneal single application . Blood samples in the second, and then 10º, 18º and 26º days after treatment were collected in order to perform WBC, proteins and electrophoretic fractionation of proteins. Regarding the healing activity, also 48 mice divided into six groups were used, in whom surgical wounds in the size of 1cm2 were purposely inflicted , after the dissociative anesthesia were applied . Two groups served as control ones, for clean and infected wounds. Two groups of S. aureus with infected wounds and two groups with clean sores were treated with 5% and 10 % propolis, being this concentration choice based on a pilot study previously performed. The results showed that treatment with propolis influences leukocyte and protein concentrations in a dose- dependent manner, with the highest dose triggering leukocytosis with lymphocytosis and increasing the protein fraction of gamaglobulínica, on the 10th day after the start of treatment. It also indicated that the concentration of the solution influence the time of healing of infected wounds, since the process on the group treated with 5% solution happened faster.


Subject(s)
Animals , Male , Female , Mice , Propolis/adverse effects , Staphylococcus aureus/isolation & purification , Wound Healing , Mice/classification , Leukocytosis/classification
3.
Clin Exp Immunol ; 173(2): 242-9, 2013 Aug.
Article in English | MEDLINE | ID: mdl-23607333

ABSTRACT

Genetic polymorphism studies of cytokines may provide an insight into the understanding of acute kidney injury (AKI) and death in intensive care unit (ICU) patients. The aim of this study was to investigate whether the genetic polymorphisms of -308 G < A tumour necrosis factor (TNF)-α, -174 G > C interleukin (IL)-6 and -1082 G > A IL-10 may predispose ICU patients to the development of AKI and/or death. In a prospective nested case-control study, 303 ICU patients and 244 healthy individuals were evaluated. The study group included ICU patients who developed AKI (n = 139) and 164 ICU patients without AKI. The GG genotype of TNF-α (low producer phenotype) was significantly lower in the with AKI than without AKI groups and healthy individuals (55 versus 62 versus 73%, respectively; P = 0·01). When genotypes were stratified into four categories of TNF-α/IL-10 combinations, it was observed that low TNF-α plus low IL-10 producer phenotypes were more prevalent in patients with AKI, renal replacement therapy and death (P < 0·05). In logistic regression analysis, low TNF-α producer plus low IL-10 producer phenotypes remained as independent risk factors for AKI and/or death [odds ratio (OR) = 2·37, 95% confidence interval (CI): 1·16-4·84; P = 0·02] and for renal replacement therapy (RRT) and/or death (OR = 3·82, 95% CI: 1·19-12·23; P = 0·02). In this study, the combination of low TNF-α plus low IL-10 producer phenotypes was an independent risk factor to AKI and/or death and RRT and/or death in critically ill patients. Our results should be validated in a larger prospective study with long-term follow-up to emphasize the combination of these genotypes as potential risk factors to AKI in critically ill patients.


Subject(s)
Acute Kidney Injury/diagnosis , Acute Kidney Injury/immunology , Interleukin-10/metabolism , Tumor Necrosis Factor-alpha/metabolism , Acute Kidney Injury/genetics , Acute Kidney Injury/mortality , Aged , Aged, 80 and over , Case-Control Studies , DNA Mutational Analysis , Female , Genetic Predisposition to Disease , Humans , Intensive Care Units , Interleukin-10/genetics , Interleukin-6/genetics , Interleukin-6/metabolism , Male , Middle Aged , Polymorphism, Genetic , Prognosis , Prospective Studies , Survival Analysis , Tumor Necrosis Factor-alpha/genetics
4.
Curr Med Chem ; 19(13): 2051-8, 2012.
Article in English | MEDLINE | ID: mdl-22420337

ABSTRACT

Schistosomiasis, caused by blood flukes of the genus Schistosoma, still imposes a considerable public health burden on large parts of the world. The control of this disease depends almost exclusively on the drug praziquantel, and there are no alternative drugs in sight. Natural compounds have recently attracted significant attention due to their relevance to parasitic infection and potential development into new therapeutic agents. Epiisopiloturine is an imidazole alkaloid isolated from the leaves of Pilocarpus microphyllus (Rutaceae), a native plant from Brazil. Here, we report the in vitro effect of this drug on the survival time of Schistosoma mansoni of different ages, such as 3 h old and 1, 3, 5, and 7 days old schistosomula, 49-day-old adults, and on egg output by adult worms. Epiisopiloturine at a concentration of 300 µg/mL caused the death of all schistosomula within 120 h. Extensive tegumental alterations and death were observed when adult schistosomes had been exposed to 150 µg/mL of the epiisopiloturine. At the highest sub-lethal dose of alkaloid (100 µg/mL), a 100% reduction in egg laying of paired adult worms was observed. Additionally, epiisopiloturine showed selective antischistosomal activity and exhibited no cytotoxicity to mammalian cells. This report provides the first evidence that epiisopiloturine is able to kill S. mansoni of different ages and inhibit worm egg laying.


Subject(s)
4-Butyrolactone/analogs & derivatives , Imidazoles/pharmacology , Schistosoma mansoni/drug effects , 4-Butyrolactone/chemistry , 4-Butyrolactone/isolation & purification , 4-Butyrolactone/pharmacology , 4-Butyrolactone/toxicity , Animals , Chlorocebus aethiops , Imidazoles/chemistry , Imidazoles/isolation & purification , Imidazoles/toxicity , Mice , Pilocarpus/chemistry , Reproduction/drug effects , Schistosoma mansoni/physiology , Vero Cells
5.
Arq. bras. med. vet. zootec ; 63(3): 757-760, June 2011. tab
Article in Portuguese | LILACS | ID: lil-595597

ABSTRACT

The prevalence of anti-lentiviruses antibodies of small ruminants was investigated in goat herds in the city of Teresina, PI, Brazil. A seroepidemiological survey was conducted involving 480 animals, apparently healthy, belonging to six rural properties. The diagnostic test was the agar gel immunodiffusion (AGID), using antigens produced from cellular cultures infected with caprine arthritis encephalitis virus (CAEV Cork). Prevalences by gender and age were estimated considering sampling fractions for each farm. A general prevalence of 4.2 percent, was observerved, being 4.2 percent for females and 3.6 percent for males. Prevalences were higher among older goats. Regarding the breed standard, 23.5 percent were of the Anglo Nubian, 5.9 percent of the Boer, 35.3 percent Anglo Nubian x Boer crossbred, and 35.3 percent of undefined breed. It is concluded that small ruminant lentiviruses are endemic among goat herds of Teresina.


Subject(s)
Animals , Arthritis-Encephalitis Virus, Caprine , Lentivirus Infections/epidemiology , Lentivirus Infections/immunology , Lentivirus Infections/prevention & control , Lentivirus Infections/transmission , Lentivirus Infections/veterinary , Lentivirus , Seroepidemiologic Studies
6.
J Hum Hypertens ; 25(8): 492-9, 2011 Aug.
Article in English | MEDLINE | ID: mdl-20827284

ABSTRACT

Among hypertensive patients, cardiovascular disease morbidity is common, even in those who are adequately treated. New pharmacological strategies to mitigate the burden of arterial hypertension are needed. This 12-month, randomized, double-blind placebo-controlled study investigated the effect of statin (fluvastatin) treatment on ambulatory blood pressure (ABP), exercise blood pressure (EBP), myocardial structure, endothelial function and insulin resistance in 50 hypertensive patients. At baseline, the groups were comparable in terms of demographic characteristics, ABP, EBP, endothelial function and homeostasis model assessment of insulin resistance (HOMA-IR). At the end of the study, there was no difference between groups in terms of resting systolic blood pressure. However, maximum systolic EBP was lower in the treatment group than in the placebo group (175 ± 18 vs 192 ± 23 mm Hg, P<0.05), as was left ventricular mass index (LVMI; 82 ± 15 vs 100 ± 23, P<0.05), and HOMA-IR index was lower after fluvastatin treatment (2.77 ± 1.46 vs 3.33 ± 1.73, P<0.05). Changes in lipid profile were not correlated with blood pressure, endothelial function, LVMI or HOMA-IR data. In hypertensive patients, fluvastatin can improve maximum systolic EBP, myocardial remodelling and insulin resistance, independently of lipid profile variations and endothelial function.


Subject(s)
Fatty Acids, Monounsaturated/therapeutic use , Hydroxymethylglutaryl-CoA Reductase Inhibitors/therapeutic use , Hypertension/drug therapy , Indoles/therapeutic use , Insulin Resistance , Myocardium/pathology , Adult , Blood Pressure/drug effects , Double-Blind Method , Female , Fluvastatin , Humans , Hypertension/metabolism , Hypertension/pathology , Male , Middle Aged
7.
Transplant Proc ; 42(4): 1084-9, 2010 May.
Article in English | MEDLINE | ID: mdl-20534229

ABSTRACT

The aim of this study was to investigate whether slow graft function (SGF) after transplantation of deceased-donor kidneys affected the prevalence of anemia or the glomerular filtration rate (GFR). We retrospectively evaluated the records of 534 kidney transplant patients who were classified based on their initial renal function, namely, immediate graft function (IGF), slow graft function (SGF), or delayed graft function (DGF). Among the 534 kidney transplant patients studied, the occurrences of each condition were IGF 104, SGF 133, and DGF 297. Six months after transplantation, a greater percentage of DGF patients were anemic compared with the others (P = .028). However, at 12 months after transplantation, SGF patients showed more anemia than the IGF group. DGF and SGF patients displayed similar GFR values at 18 and 24 months after transplantation. However, IGF patients displayed higher GFRs, even when subjects who experienced acute rejection episodes were censored (P = .004). The incidence of acute rejection episodes was similar among SGF and DGF patients. Patients displaying SGF after deceased-donor transplantation showed a greater tendency to be anemic than those displaying IGF. This study also suggested that SGF patients were at risk for acute rejection episodes and/or significantly reduced kidney function as measured by GFR.


Subject(s)
Glomerular Filtration Rate , Kidney Transplantation/physiology , Anemia/epidemiology , Cadaver , Delayed Graft Function/epidemiology , Follow-Up Studies , Graft Survival , Humans , Immunosuppressive Agents/adverse effects , Immunosuppressive Agents/therapeutic use , Kidney Function Tests , Kidney Transplantation/adverse effects , Kidney Transplantation/immunology , Postoperative Complications/epidemiology , Renal Dialysis , Retrospective Studies , Time Factors , Tissue Donors , Treatment Failure
8.
Clin Nephrol ; 73(1): 7-13, 2010 Jan.
Article in English | MEDLINE | ID: mdl-20040346

ABSTRACT

BACKGROUND: Soluble Fas levels (sFas) are increased in the serum of uremic patients and are associated with the presence of anemia and recombinant human EPO (rHuEPO) dosage in dialysis patients. It is possible that sFas levels are associated with an increased need for serum erythropoietin levels (Epo) in chronic kidney disease and dialysis patients in order to maintain hematocrit (Hct) levels. AIMS: To investigate the relationship between serum sFas levels, serum Epo levels and the ratio between Epo levels and Hct in uremic patients. METHODS: We studied 52 predialysis chronic kidney disease patients (CKD; 33 M, 57 +/- 12 years, hematocrit (Hct) = 37 +/- 7%), 29 peritoneal dialysis patients (PD; 12 M, 54 +/- 14 years, Hct = 36 +/- 7%), 29 hemodialysis patients (HD; 19 M, 47 +/- 14 years, Hct = 33 +/- 5%) and 29 healthy volunteers (control group 17 M, 50 +/- 16 years, Hct = 43 +/- 3%). We examined the relationship between Hct and serum levels of Epo, sFas, C-reactive protein, IL-6 and iron status. The ratio of serum Epo divided by Hct (Epo/Hct) was used as an indicator of Epo responsiveness. RESULTS: Compared to normal subjects, the CKD, PD and HD groups presented lower Hct levels and higher serum levels of sFas, Epo, Epo/Hct and IL-6. Serum levels of sFas correlated negatively with albumin (r = -0.24, p = 0.02), IL-6 (r = -0.18, p = 0.04) and Epo/Hct (r = -0.37, p < 0.001). In multivariate analysis, after adjusting for markers of iron store and inflammation, only sFas correlated with Epo/Hct. In the CKD group, there were negative correlations between serum levels of sFas and glomerular filtration rate (GFR) (r = -0.45, p < 0.001) and between Epo/Hct and GFR (r = -0.32; p = 0.02). There was a positive correlation between Epo/Hct and serum levels of sFas in the CKD group (r = 0.31, p = 0.03) and in the HD groups (r = 0.58, p = 0.001). CONCLUSION: Our findings show that serum sFas is associated with higher Epo/Hct ratio, suggesting that sFas may be a marker of Epo hyporesponsiveness in uremia. Further studies are needed to determine whether sFas is just a marker of Epo hyporesponsiveness or is also involved in its pathophysiology.


Subject(s)
Erythropoietin/blood , Fas Ligand Protein/blood , Inflammation/blood , Kidney Failure, Chronic/blood , Adult , Aged , Analysis of Variance , Anemia, Iron-Deficiency/blood , Anemia, Iron-Deficiency/complications , C-Reactive Protein/metabolism , Chi-Square Distribution , Female , Humans , Interleukin-6/blood , Iron/blood , Kidney Failure, Chronic/complications , Male , Middle Aged , Patient Selection , Regression Analysis , Renal Dialysis
9.
Transplant Proc ; 41(4): 1210-3, 2009 May.
Article in English | MEDLINE | ID: mdl-19460519

ABSTRACT

Delayed graft function (DGF) is defined as the necessity for dialysis during the first week after transplantation. This study sought to describe patterns of dialysis prescription and evaluate the impact of dialysis dose in acute rejection. Among 82 patients who received a deceased donor kidney transplant, clinical and laboratory data were evaluated at the moment of dialysis indication. Prescribed and delivered dialysis doses (Kt/V and urea reduction ratio) were analyzed during the first dialysis and the first week (Kt/V) after transplantation. We examined the association between Kt/V and acute rejection. Prescribed Kt/V at the first dialysis session was adequate (2.24 +/- 0.51). However, delivered Kt/V was inadequate (0.75 +/- 0.38). Prescribed and delivered Kt/V during the first week after transplantation were suboptimal, namely, 2.45 +/- 1.52 and 1.56 +/- 0.99, respectively. Dialysis dose had no impact on the occurrence of an acute rejection episode. Among DGF patient, dialysis was prescribed late and a low dose was achieved.


Subject(s)
Delayed Graft Function/therapy , Graft Rejection/prevention & control , Kidney Transplantation , Renal Dialysis , Adult , Aged , Female , Fluid Therapy , Humans , Male , Middle Aged , Patient Selection , Postoperative Care , Transplant Recipients , Urea
10.
Arq. bras. med. vet. zootec ; 60(5): 1123-1127, out. 2008. tab
Article in Portuguese | LILACS | ID: lil-500079

ABSTRACT

Determinaram-se os perfis do hemograma e do proteinograma de 24 cutias (Dasyprocta sp.) saudáveis, criadas em cativeiro, e avaliaram-se a influência de sexo e de idade e a interação sexo-idade. O delineamento foi inteiramente ao acaso em arranjo fatorial 2 x 4 (dois sexos e quatro faixas etárias), com três repetições. Os valores do hemograma e do proteinograma foram: leucócitos totais: 7,24±2,52 (x10³/µl), basófilos: 0,11±0,2 (x10³/µl), eosinófilos: 0,32±0,2 (x10³/µl), segmentados: 2,5±1,22 (x10³/µl), linfócitos: 3,58±1,52 (x10³/µl), monócitos: 0,76±0,47 (x10³/µl), hemácias: 6,51±0,83 (x10(6)/µl), hematócrito: 43,58±3,41 por cento, hemoglobina: 12,71±1,04g/dl, volume corpuscular médio: 67,61±7,52ml, concentração de hemoglobina corpuscular média: 29,19±1,61 por cento, plaquetas: 150,08±63,39 (x10³/ml), proteínas totais: 6,04±1,77g/dl, albumina: 2,07±0,79g/dl e globulinas: 3,97±1,54g/dl. Os parâmetros relativos à série vermelha e ao proteinograma não diferiram quanto ao sexo ou idade, e não houve interação sexo-idade. Os valores de linfócitos foram mais baixos entre os animais mais velhos.


The profiles of hemogram and proteinogram of healthy agouti (Dasyprocta sp.) grown in captivity, in Piauí State, Brazil, were determined, as well as the influence of gender and age factors and the relationship between sex-age them. A completely randomized design was used in a factorial arrange 2x4, two genders and four ages, totaling 24 animals. The mean values of the hemogram were: erythrocytes = 6.51±0.83 (x10(6)); hematocrit = 43.58±3,41 percent; hemoglobin = 12.71±1,04g/dl; mean corpuscular volume = 64.61±7.52μl; mean corpuscular hemoglobin concentration = 29.19±1,61 percent; platelets= 150.08±63.39 (x10³/μl); white blood cells = 7.24±2.52 (x10³/μl); basophils = 0.11±0,2 (x10³/μl); eosinophils = 0,32±0,2 (x10³/μl); neutrophils = 2.5±1,22 (x10³/μl); lymphocytes = 3.58±1.52 (x10³/μl); and monocytes = 0.76±0.47 (x10³/μl). The mean values of the proteinogram were: total serum proteins = 6.04±1.77g/dl; albumin = 2.07±0.79g/dl and globulin = 3.97±1.54g/dl. Neither gender nor age influenced the results of the studied parameters. Association between gender and age was not verified. The values of lymphocytes were lower in older animals.


Subject(s)
Animals , Blood Cell Count/methods , Proteins/analysis , Rodentia
11.
Arq. bras. med. vet. zootec ; 60(3): 651-655, jun. 2008. tab
Article in Portuguese | LILACS | ID: lil-487910

ABSTRACT

Determinou-se o perfil do ionograma e enzimas hepáticas de cutias (Dasyprocta sp.) saudáveis, criadas em cativeiro, como também se avaliou a influência de sexo, idade e interação sexo-idade. Foi adotado um delineamento inteiramente ao acaso, em arranjo fatorial 2 x 4 (dois sexos e quatro faixas etárias), com três repetições, totalizando 24 cutias. Foram determinados os valores para o cálcio (Ca), fósforo (P), cloretos (Cl), aspartato aminotransferase (AST), alanina aminotransferase (ALT), fosfatase alcalina (FA) e gama-glutamiltransferase (GGT). Os valores médios obtidos foram: Ca= 7,62+2,59mg/dl; P= 3,91+1,41mg/dl; Cl= 58,63+16,45mg/dl; AST= 119,54+79,35UI/ml; ALT= 28,08+15,53UI/ml; FA= 26,95+14,01UI/ml e GGT= 25,34+19,44UI/ml. O valor de P foi maior nas fêmeas e da FA nos machos. Os níveis de FA diminuíram com o aumento da idade.


This research studied the profile of the ionogram and hepatic enzymes of healthy agoutis (Dasyprocta sp.) raised in captivity as well as evaluated the influence of gender, age and interaction gender-age. It was used a completely randomized design, in a factorial arrangement of 2 x 4 (two genders and four age groups), with three repetitions, totaling 24 agoutis. The values were determined for calcium (Ca), phosphorus (P), chlorides (Cl), aspartate aminotransaminase (AST), alanine aminotransaminase (ALT), alkaline phosphatase (ALP), and gamma-glutamyltransferase (GGT). The mean values were: Ca= 7.62±2.59mg/dl; P= 3.91±1.41mg/dl; Cl= 58.63±16,45mg/dl; AST= 119.54±79.35UI/ml; ALT= 28.08±15.53UI/ml; ALP= 26.95±14.01UI/ml, and GGT= 25.34±19.44UI/ml. The value of P was larger in females and ALP in males. As the age increased, levels of ALP decreased.


Subject(s)
Animals , Alanine Transaminase/analysis , Aspartate Aminotransferases/analysis , Electrolytes , Enzymes , Spectrophotometers/methods , Alkaline Phosphatase/analysis , Rodentia , gamma-Glutamyltransferase/analysis
12.
Vet Pathol ; 44(3): 314-9, 2007 May.
Article in English | MEDLINE | ID: mdl-17491072

ABSTRACT

Conidiobolomycosis is reported in the state of Piauí, in the semiarid region of northeastern Brazil. Affected sheep had depression, weight loss, serous or mucohemorrhagic nasal discharge, and cranium-facial asymmetry from exophthalmos of 1 eye, generally with increased volume of the eyeball, keratitis, and corneal ulceration. At necropsy of 60 sheep, friable masses were observed in the posterior region of the nasal cavity, often destroying the ethmoturbinate bones. Frequently, the lesions invaded the nasal sinuses, cribiform plate, orbit, and brain. The masses were irregular, granular with moist surfaces, and soft and friable with white, yellow, or tan coloration. Dissemination of the lesion to lungs was observed in 27 sheep, to the brain in 26, to lymph nodes in 3, to the kidney in 2, and to the gallbladder and heart in 1. The microscopic examination showed granulomatous inflammation composed of central necrosis surrounded by lymphocytes, epithelioid and giant cells, and fibrous tissue. In all lesions, negatively stained structures representing hyphae were surrounded by Splendore-Hoeppli material. Coagulative necrosis, thrombosis, and vasculitis were also observed. Grocott methenamine silver stain showed 8-30-microm-thick hyphae, rarely septate or ramified, irregular in shape, and with black contoured wall, sometimes with bulbous dilatation in the extremities. On electron microscopy, the hyphae had a thick double wall surrounded by cellular remnants and an inflammatory exudate. Conidiobolus coronatus was isolated from the lesions of 6 sheep. Conidiobolomycosis is an important disease of sheep in the state of Piauí, and other regions of northeastern Brazil.


Subject(s)
Conidiobolus/isolation & purification , Sheep Diseases/microbiology , Sheep/microbiology , Zygomycosis/veterinary , Animals , Brazil/epidemiology , Exophthalmos/microbiology , Exophthalmos/pathology , Exophthalmos/veterinary , Head/pathology , Lung/pathology , Sheep Diseases/epidemiology , Sheep Diseases/pathology , Zygomycosis/epidemiology , Zygomycosis/pathology
13.
Fertil Steril ; 73(2): 280-3, 2000 Feb.
Article in English | MEDLINE | ID: mdl-10685529

ABSTRACT

OBJECTIVE: To search for germline activating mutations of the FSH receptor in girls with gonadotropin-independent precocious puberty. DESIGN: Molecular studies in human tissue. SETTING: Four girls with polycystic ovaries and gonadotropin-independent isosexual precocious puberty without clinical and molecular features of McCune-Albright syndrome. INTERVENTION(S): Peripheral blood was used for DNA extraction. The alpha-subunit of the Gs gene and the entire exon 10 of FSH receptor gene were amplified by polymerase chain reaction (PCR). Gs-alpha mutations characteristic of McCune-Albright syndrome were excluded by denaturating gradient gel electrophoresis (DGGE) and allele-specific PCR. Exon 10 of the FSH receptor gene was analyzed by DGGE and direct sequencing. MAIN OUTCOME MEASURE(S): Results of DGGE and direct sequencing. RESULT(S): No germline activating mutations were detected in exon 10 of our patients. Instead, two previously described polymorphisms were found, leading to the substitution of alanine for threonine at position 307 and of serine for asparagine at position 680 of the FSH receptor molecule. CONCLUSION(S): Germline activating mutations were not found in exon 10 of the FSHR gene in any of our patients. Further studies, preferably in ovarian tissue, will be required to exclude the presence of somatic activating mutations of the FSH receptor in these patients.


Subject(s)
Germ-Line Mutation , Ovarian Cysts/genetics , Puberty, Precocious/genetics , Receptors, FSH/genetics , Amino Acid Substitution , Child , Child, Preschool , Electrophoresis/methods , Female , Heterozygote , Humans
14.
J Clin Endocrinol Metab ; 84(10): 3539-44, 1999 Oct.
Article in English | MEDLINE | ID: mdl-10522992

ABSTRACT

To establish normative data and determine the value of fluorometric AutoDELFIA assays (Wallac Oy) in the investigation of precocious puberty, we determined serum levels of LH, FSH, testosterone, and estradiol under basal and GnRH-stimulated conditions in 277 normal subjects at various pubertal stages and in 77 patients with precocious puberty. A substantial overlap was observed in basal and GnRH-stimulated gonadotropin levels in normal individuals of both sexes with pubertal Tanner stages 1 and 2. The 95th percentile of the normal prepubertal population was the cut-off limit between prepubertal and pubertal levels. These limits were 0.6 IU/L in both sexes for basal LH, 9.6 IU/L in boys and 6.9 IU/L in girls for peak LH after GnRH stimulation, 19 ng/dL in boys for basal testosterone, and 13.6 pg/mL in girls for basal estradiol. Basal and peak LH exceeding these limits were considered positive tests for the diagnosis of gonadotropin-dependent precocious puberty. According to these criteria, the sensitivities of basal and peak LH for the latter diagnosis were 71.4% and 100% in boys, and 62.7% and 92.2% in girls. The specificity and positive predicted value were 100% in both sexes for basal and peak LH levels. The negative predicted values for basal and peak LH were 62.5% and 100% in boys, and 40.6% and 76.5% in girls. Basal and GnRH-stimulated FSH levels overlapped among the various pubertal stages in normal subjects and were, in general, not helpful in the differential diagnosis of precocious puberty. In conclusion, basal LH levels were sufficient to establish the diagnosis of gonadotropin-dependent precocious puberty in 71.4% of boys and 62.7% of girls. In the remaining patients, a GnRH stimulation test was still necessary to confirm this diagnosis. Finally, suppressed LH and FSH levels after GnRH stimulation indicate gonadotropin-independent sexual steroid production.


Subject(s)
Fluorometry/standards , Puberty, Precocious/diagnosis , Adolescent , Adult , Child , Child, Preschool , Female , Gonadotropin-Releasing Hormone , Gonadotropins/blood , Humans , Male , Puberty, Precocious/blood , Reference Values , Steroids/blood
15.
Fertil Steril ; 71(4): 597-601, 1999 Apr.
Article in English | MEDLINE | ID: mdl-10202864

ABSTRACT

OBJECTIVE: To review clinical findings, hormone levels, and DNA analyses in genetic males and females with inactivating mutations of the LH receptor gene. DESIGN: Review of reported cases. SETTING: A university hospital. PATIENT(S): Genetic males and females with inactivating mutations of the LH receptor gene. RESULT(S): The clinical presentation in genetic males ranged from female genitalia to male genitalia with micropenis caused by Leydig cell hypoplasia. Genetic females presented with amenorrhea or oligomenorrhea, enlarged cystic ovaries, and infertility. Both males and females had elevated LH levels and LH/FSH ratios. Sequencing of genomic DNA revealed homozygous or compound heterozygous deletions, nonsense mutations, or missense mutations in the LH receptor gene. CONCLUSION(S): This study of patients with inactivating mutations of the LH receptor indicates that in genetic males, the action of hCG and LH is necessary for the normal development of primary and secondary sexual characteristics. In contrast, secondary sexual characteristics develop in genetic females in the absence of LH action, but they fail to ovulate.


Subject(s)
Infertility/genetics , Menstruation Disturbances/genetics , Mutation , Receptors, LH/genetics , Female , Humans , Male , Receptors, FSH/genetics
16.
Clin Endocrinol (Oxf) ; 51(6): 701-7, 1999 Dec.
Article in English | MEDLINE | ID: mdl-10619974

ABSTRACT

OBJECTIVE: To review clinical and hormonal characteristics of new and published females with LH resistance. PATIENTS: Seven sisters of patients with male pseudohermaphroditism due to LH resistance. MEASUREMENTS: Clinical characteristics, hormonal levels, pelvic ultrasound and molecular studies. RESULTS: Patients had: (1) normal female external genitalia; (2) spontaneous breast and pubic hair development at ages 9-13 years; (3) menarche at 12-20 years, followed by irregular menstrual cycles (3 weeks to 15 months); (4) infertility; (5) withdrawal bleeding after progesterone administration; (6) elevated serum LH levels (10-38 IU/l, normal 0. 9-8.4) and elevated LH/FSH ratio with normal androgen levels; (7) low or normal oestradiol levels for the follicular phase; and (8) normal or small uterus and normal or increased ovary size with one or more cysts at ultrasound examination. Ovarian biopsy showed antral follicles and confirmed lack of ovulation. One homozygous mutation in exon 11 of the LH receptor gene was found in each of 4 patients: 1 nonsense (Arg554Stop) and 2 missense mutations (Glu354Lys and Ala593Pro) and 1 microdeletion (Leu-608, Val-609) were found. In 3 patients (1 family) with the same phenotypic characteristics, no mutations in the coding sequence of the LH receptor gene were found. CONCLUSIONS: We conclude that women with LH resistance have spontaneous breast development, primary or secondary amenorrhoea, infertility, elevated serum LH levels and LH/FSH ratio with normal androgen levels and normal or enlarged cystic ovaries. Therefore, in females, primary and secondary sexual characteristics develop independently of LH action. However, LH stimulation is necessary for normal ovarian steroidogenesis and ovulation.


Subject(s)
Disorders of Sex Development/blood , Luteinizing Hormone/blood , Adolescent , Adult , Amenorrhea/blood , Amenorrhea/genetics , Amenorrhea/pathology , Disorders of Sex Development/genetics , Disorders of Sex Development/pathology , Estradiol/blood , Female , Follicle Stimulating Hormone/blood , Gene Deletion , Humans , Infertility, Female/blood , Infertility, Female/genetics , Infertility, Female/pathology , Mutation , Ovary/diagnostic imaging , Ovary/pathology , Phenotype , Progesterone , Receptors, LH/genetics , Ultrasonography , Uterus/diagnostic imaging
17.
Fertil Steril ; 70(3): 565-7, 1998 Sep.
Article in English | MEDLINE | ID: mdl-9757892

ABSTRACT

OBJECTIVE: To investigate the presence of FSH receptor gene mutations in women with premature ovarian failure (POF). DESIGN: Clinical and molecular studies. SETTING: Research laboratory in a university setting. PATIENT(S): Fifteen 46,XX women with POF and 42 normal fertile controls. INTERVENTION(S): Exon 7 was amplified and digested with BsmI to screen for the previously described inactivating mutation C566T. Exon 10 was screened for mutations by denaturing gradient gel electrophoresis and direct sequencing. MAIN OUTCOME MEASURE(S): Polymerase chain reaction followed by restriction enzyme analysis, denaturing gradient gel electrophoresis, and direct sequencing. RESULT(S): No inactivating mutations were identified in exons 7 and 10 of the FSH receptor gene in women with familial or sporadic POF. Exon 10 had two polymorphisms, G919A and G2039A, whose allelic frequencies were 46.7% and 56.6%, respectively, in women with POF. The allelic frequency of both polymorphisms was 59.5% in normal fertile controls. CONCLUSION(S): No inactivating mutations in exons 7 and 10 of the FSH receptor gene were identified in Brazilian women with POF. A high frequency of two polymorphisms that are in linkage disequilibrium was found in exon 10 of this gene.


Subject(s)
Periodicity , Primary Ovarian Insufficiency/genetics , Receptors, FSH/genetics , Adolescent , Adult , Brazil , Case-Control Studies , Female , Humans , Mutation , Polymerase Chain Reaction
18.
Fertil Steril ; 67(2): 394-7, 1997 Feb.
Article in English | MEDLINE | ID: mdl-9022621

ABSTRACT

OBJECTIVE: To report the clinical, hormonal, and histopathological features of a woman with ovarian resistance to LH. DESIGN: Clinical study. SETTING: University hospital. PATIENT(S): A woman with amenorrhea, sister of a patient with male pseudohermaphroditism due to Leydig cell hypoplasia. INTERVENTION(S): Blood drawing before and after GnRH stimulation and also after dexamethasone and hCG administration, pelvic ultrasound, and ovarian biopsy. MAIN OUTCOME MEASURE(S): Karyotype, gonadotropin and steroid measurements, follicular diameter, ovarian histology, and sequencing of the LH receptor gene. RESULT(S): Patient had normal female external genitalia, normal breast development at puberty, rare episodes of vaginal bleeding, and infertility. The karyotype was 46,XX. She had elevated serum LH levels, whereas E2 and P concentrations were in the range seen in the early follicular phase. Pelvic ultrasound revealed a slightly hypoplastic uterus and enlarged polycystic ovaries. A normal follicular reserve for age, antral follicles, and absence of corpora lutea or albicans were observed on ovarian biopsy. Exon 11 of the LH receptor gene had a normal sequence. CONCLUSION(S): In our patient with ovarian resistance to LH, FSH stimulated follicular development until the preovulatory stage, but E2 levels remained in the early follicular phase range, still sufficient for normal pubertal feminization. Apparently, LH is necessary for ovulation and corpus luteum formation.


Subject(s)
Amenorrhea/etiology , Infertility, Female/etiology , Luteinizing Hormone/physiology , Ovary/physiology , Adult , Amenorrhea/genetics , DNA/genetics , Disorders of Sex Development/genetics , Drug Resistance , Female , Gonadotropins/blood , Humans , Infertility, Female/genetics , Karyotyping , Male , Ovarian Follicle/pathology , Ovary/pathology , Polycystic Ovary Syndrome/diagnostic imaging , Receptors, LH/genetics , Steroids/blood , Ultrasonography , Uterus/diagnostic imaging
19.
J Hypertens ; 15(12 Pt 1): 1397-402, 1997 Dec.
Article in English | MEDLINE | ID: mdl-9431844

ABSTRACT

BACKGROUND: Apparent mineralocorticoid excess (AME) is a cause of low-renin, low-aldosterone hypertension in which cortisol acts as a mineralocorticoid. The condition reflects an inability to inactivate cortisol to cortisone due to defective activity of the type 2 isozyme of 11beta-hydroxysteroid dehydrogenase (11beta-HSD2). Homozygous mutations in 11beta-HSD2 gene in patients with AME have been described. A 7-year-old Brazilian girl had previously been found to have AME. Her father recently presented with mineralocorticoid hypertension at age 38 years. OBJECTIVE: To describe the clinical details, to perform steroid analyses and to assess the molecular basis for the hypertension in this kindred. METHODS: The 11beta-HSD2 gene was amplified from genomic DNA by the polymerase chain reaction and sequenced by direct chain-termination sequencing on an automatic DNA sequencer. The sequencing results were validated by restriction-site polymorphism. The mutant 11beta-HSD2 protein was expressed in Chinese hamster ovary polyoma cells and enzymatic activity was assessed by metabolizing cortisol in vitro. RESULTS: Sequence analysis of genomic DNA revealed a novel C1061T point mutation in exon V of the human 11beta-HSD2 gene, resulting in an amino acid substitution of alanine by valine at codon 328 of the enzyme protein (A328V). Expression studies confirmed that the mutant protein was devoid of 11beta-HSD2 activity. A HhaI restriction-site polymorphism confirmed that the proband was homozygous for the mutation whereas both parents were heterozygotes. The father of the proband had hypertension, a normal serum potassium level, suppressed plasma renin activity and plasma aldosterone level and a moderately elevated urinary cortisol: cortisone metabolite ratio. CONCLUSIONS: AME in this kindred is caused by a novel mutation in the 11beta-HSD2 gene. Detection of hypokalaemia, at least in this kindred, is an insensitive screening test for mineralocorticoid-based hypertension. In contrast to results from previously investigated kindreds, we have demonstrated that this kindred has an abnormal phenotype in the heterozygote state. Further studies are now required in order to evaluate the role of 11beta-HSD2 activity in the pathophysiology of 'essential' hypertension.


Subject(s)
Heterozygote , Hypertension/genetics , Mineralocorticoids/metabolism , 11-beta-Hydroxysteroid Dehydrogenases , Adult , Animals , Base Sequence , Brazil , Child , Cricetinae , Cricetulus , Female , Humans , Hydroxysteroid Dehydrogenases/genetics , Hydroxysteroid Dehydrogenases/metabolism , Ovarian Neoplasms/metabolism , Ovarian Neoplasms/pathology , Point Mutation , Tumor Cells, Cultured
20.
Fertil Steril ; 65(3): 495-502, 1996 Mar.
Article in English | MEDLINE | ID: mdl-8774276

ABSTRACT

OBJECTIVE: To investigate whether luteal and endometrial abnormalities occur more frequently in an infertile population and thus contribute to infertility. DESIGN: Prospective controlled clinical study. SETTING: Outpatient clinic in an academic research institution. PARTICIPANTS: Thirty-three fertile controls and 31 infertile women without ovulatory disorders, tubal disease, or male factors. INTERVENTIONS: All women underwent an endometrial biopsy 9 days after the LH surge followed by an IM injection of 5,000 IU hCG. Blood samples were drawn immediately before hCG administration for serum P and placental protein 14 (PP14) measurements, at 6 hours after hCG stimulation for serum P concentrations, and on day 5 after hCG administration for serum PP14 levels. MAIN OUTCOME MEASURES: Histologic dating of the endometrium and serum P and PP14 measurements. RESULTS: Abnormal endometrial biopsies occurred more frequently in infertile (43%) than in fertile women (9%). Except for one case, these specimens were not associated with low hCG-stimulated P levels. Serum PP14 measurements varied widely and did not discriminate subjects with abnormal endometrial development. CONCLUSIONS: Disruption of endometrial maturation without a concomitant defect of the corpus luteum occurs more frequently in an infertile population and thus may contribute to infertility.


Subject(s)
Corpus Luteum/pathology , Endometrium/pathology , Glycoproteins/blood , Infertility, Female/blood , Infertility, Female/pathology , Pregnancy Proteins/blood , Progesterone/blood , Adult , Biopsy , Chorionic Gonadotropin , Endometrium/physiopathology , Female , Glycodelin , Humans , Infertility, Female/physiopathology , Menstrual Cycle , Middle Aged , Prospective Studies , Time Factors
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