Impacto da terapia hormonal sobre o peso corpóreo

OBJETIVO: avaliar o efeito da terapêutica hormonal (TH) no peso de mulheres na peri-menopausa, assim como o efeito de diferentes regimes terapêuticos no referido parâmetro. MÉTODOS: estudo retrospectivo de 139 mulheres, com menopausa há menos de 2 anos, acompanhadas na consulta de climatério do nosso departamento. Obtiveram-se dois grupos: mulheres a quem se iniciou TH (n=89) e outro, grupo controle, sem terapia hormonal (n=50). Em cada grupo, foi avaliada a modificação ponderal no intervalo de 1 ano após a primeira consulta. Nas submetidas a TH, avaliou-se esse mesmo parâmetro em função de diferentes regimes terapêuticos preconizados: estrogênio isolado vs estroprogestagênio e dose standard vs baixa dosagem. A análise estatística foi realizada com recurso ao programa SPSS®, adotando-se como nível de significância valores p<0,05. RESULTADOS: os dois grupos foram semelhantes no que diz respeito a características basais e demográficas. No nosso estudo, constatou-se um aumento ponderal médio superior no grupo controle relativamente ao grupo de mulheres sob TH (434 vs 76 g), embora a diferença verificada não seja estatisticamente significativa (p=0,406); Nas usuárias de TH, aquelas sob estrogênio isolado tiveram um aumento ponderal acrescido face a mulheres sob terapêutica com estroprogestativo (775 vs 24 g), embora com diferenças não significativas, o mesmo sucedendo quando analisada a dosagem de TH inicialmente prescrita (92 vs 49 g). CONCLUSÕES: apesar da crença comum do aumento ponderal associado à TH, os resultados do estudo descrito parecem contrariar esse aspecto, não havendo um ganho ponderal adicional ao normalmente associado a este período da vida da mulher.

Sintomas climatéricos e estado nutricional de mulheres na pós-menopausa usuárias e não usuárias de terapia hormonal

OBJETIVO: Analisar os sintomas climatéricos e estado nutricional em mulheres na pós-menopausa, usuárias e não usuárias de terapia hormonal (TH). MÉTODOS: Estudo analítico, exploratório, tipo inquérito populacional domiciliar, realizado na área urbana do município de Maringá, Paraná, incluindo 456 mulheres com idade entre 45 e 69 anos, no período pós-menopausa. A coleta teve como base de referência os setores censitários urbanizados (368) do município, de acordo com o Censo Demográfico Brasileiro. Foi utilizada amostra aleatória simples proporcional às mulheres residentes em cada setor censitário e, por meio de visita domiciliar, aplicou-se um questionário e verificaram-se as medidas antropométricas e pressão arterial. Para avaliação dos sintomas climatéricos, foi utilizado o Índice Menopausal de Blatt e Kupperman. A variável desfecho foi o uso de TH. RESULTADOS: A média de idade foi de 58,7 anos. O excesso de peso esteve presente em 72,6% das mulheres e a obesidade abdominal em 81,4% delas. Sintomas climatéricos de intensidade leve foram evidenciados em 69,5% das mulheres. Apenas 18,4% das mulheres faziam uso de TH e eram, na sua maioria, brancas, não fumantes, sem comorbidades e sem companheiro. Usuárias de TH apresentaram menor frequência de excesso de peso e obesidade abdominal e tiveram menor prevalência de sintomas climatéricos de intensidade severa. CONCLUSÃO: O excesso de peso e a obesidade abdominal foram prevalentes na amostra estudada. Embora em menor número, as usuárias de TH apresentaram uma frequência menor de excesso de peso e sintomas climatéricos leves e intensos na pós-menopausa.

Evaluation of an enzyme-linked immunosorbent assay to detect antibodies against Anaplasma marginale

A rapid indirect enzyme-linked immunosorbent assay (ELISA) was developed for measuring antibodies against Anaplasma marginale using a partially soluble antigen prepared from semi-purified initial bodies from erythrocytes with 80.0% of rickettsiaemia. This technique utilized alkaline phosphatase and p-nitrophenyl phosphate as reaction indicators. The high sensitivity (100.0%) was confirmed with sera from 100 calves experimentally-infected with A. marginale. All of these animals showed seroconversion before or at the same time of the first rickettsiaemia or even when it was not detected. Also the elevated specificity (94.0%) was confirmed by the low percentage of cross-reactions with sera from animals experimentally-infected with Babesia bigemina and Babesia bovis (1.4 and 6.6%, respectively). Performances of ELISA and indirect fluorescent antibody test (IFAT) with 324 sera from enzootically stable area did not show statistical difference (P>0.05), since the former showed 96.9% and the latter 97.2% of positive reactions. The advantage of this ELISA is a shorter execution time than others developed until now, allowing more samples to be analyzed.

An enzyme-linked immunosorbent assay (ELISA) for the detection of IgM antibodies against Leishmania chagasi in dogs

Visceral leishmaniasis is an emergent zoonosis with an increasing number of new cases in Brazil where the domestic dog is an important parasite reservoir in the infectious cycle of Leishmania chagasi. An enzyme-linked immunosorbent assay (ELISA), based upon the use of a total soluble antigenic preparation of L. chagasi, was adapted for the detection of IgM antibodies in the serum of infected dogs. Optimal dilutions of the antigen, using positive and negative reference sera, were determined by checkboard titrations. The specificity and sensitivity of the ELISA were 100 %. A total of 110 serum samples were taken from dogs in Belo Horizonte, Minas Gerais, Brazil, and examined for anti-L. chagasi IgM antibody by ELISA and indirect fluorescent antibody test (IFAT). About 25% (n=27) of all the dogs tested were found serologically positive for L. chagasi by IFAT, while 89.09% (n=98) were seropositive by ELISA. The results obtained by ELISA and IFAT were significantly different (P<0.01). The combined use of ELISA and IFAT is recommended in order to enable veterinary services to more efficiently detect canine visceral leishmaniasis.

The use of enzyme-linked immunosorbent assay and immunoblotting for the detection of Campylobacter fetus immunoglobulins in the cervico-vaginal mucus of female cattle

An indirect enzyme-linked immunosorbent assay was developed to detect antigen-specific secretory IgA antibodies to Campylobacter fetus subsp. venerealis in bovine vaginal mucus with a protein extract of the Campylobacter fetus subsp. venerealis by the acid glycine extraction method. Mean optical density measurement (λ=450 nm) was 0.143±0.9. The most immunoreactive protein bands of the Campylobacter fetus subsp. venerealis or Campylobacter fetus subsp. fetus recognized by IgA in immunoblotting, using bovine vaginal mucus samples, migrate at 42.6 kDa. The protein that migrates at 93 kDa was recognized exclusively for C. fetus subsp. venerealis. A positive vaginal mucus sample of a cow from negative herd recognized antigens of C. jejuni subsp. jejuni e C. fetus subsp. fetus.

Parameter estimation and use of gamma interferon assay for the diagnosis of bovine tuberculosis in Brazil

This study aimed to evaluate the interference of tuberculin test on the gamma-interferon (INFg) assay, to estimate the sensitivity and specificity of the INFg assay in Brazilian conditions, and to simulate multiple testing using the comparative tuberculin test and the INFg assay. Three hundred-fifty cattle from two TB-free and two TB-infected herds were submitted to the comparative tuberculin test and the INFg assay. The comparative tuberculin test was performed using avian and bovine PPD. The INFg assay was performed by the BovigamTM kit (CSL Veterinary, Australia), according to the manufacturer's specifications. Sensitivity and specificity of the INFg assay were assessed by a Bayesian latent class model. These diagnostic parameters were also estimate for multiple testing. The results of INFg assay on D0 and D3 after the comparative tuberculin test were compared by the McNemar's test and kappa statistics. Results of mean optical density from INFg assay on both days were similar. Sensitivity and specificity of the INFg assay showed results varying (95% confidence intervals) from 72 to 100% and 74 to 100% respectively. Sensitivity of parallel testing was over 97.5%, while specificity of serial testing was over 99.7%. The INFg assay proved to be a very useful diagnostic method.

Development and evaluation of a loop-mediated isothermal amplification (LAMP) assay for rapid detection of Actinobacillus pleuropneumoniae based the dsbE-like gene

This paper reports on the development and validation of a loop-mediated isothermal amplification assay (LAMP) for the rapid and specific detection of Actinobacillus pleuropneumoniae (A. pleuropneumoniae). A set of six primers were designed derived from the dsbE-like gene of A.pleuropneumoniae and validate the assay using 9 A. pleuropneumoniae reference/field strains, 132 clinical isolates and 9 other pathogens. The results indicated that positive reactions were confirmed for all A. pleuropneumoniae strains and specimens by LAMP at 63ºC for 60 min and no cross-reactivity were observed from other non-A.pleuropneumoniae including Haemophilus parasuis, Escherichia coli, Pasteurella multocida, Bordetella bronchiseptica, Streptococcus suis, Salmonella enterica, Staphylococcus, porcine reproductive and respiratory syndrome virus (PRRSV), and Pseudorabies virus. The detection limit of the conventional PCR was 10² CFU per PCR test tube, while that of the LAMP was 5 copies per tube. Therefore, the sensitivity of LAMP was higher than that of PCR. Moreover, the LAMP assay provided a rapid yet simple test of A. pleuropneumoniae suitable for laboratory diagnosis and pen-side detection due to ease of operation and the requirement of only a regular water bath or heat block for the reaction.

Effects of environmental modification on mastitis occurrence and hormonal changes in Holstein cows

The purpose of this research was to evaluate the effects of evaporative cooling in freestall on mastitis occurrence, milk production, and composition, as well as cortisol, T3 (triiodothyronine), and T4 (thyroxin) levels in lactating dairy cows. Twenty-eight multiparous cows averaging 70 ± 10 day postpartum were used in four treatments from January to March 2003. The treatments were: Day (cooling from 7:00 a.m. to 7:00 p.m.); Night (cooling from 7:00 p.m. to 7:00 a.m.); 24-hour (cooling 24-hour); and Control (no cooling). Wired cup test was used for clinical mastitis diagnosis, and the California Mastitis Test (CMT) was used to identify subclinical mastitis. Blood and milk samples were taken weekly for microbiological and hormonal analyses. The cortisol levels were higher than normal values in all treatment groups, suggesting stress conditions, but T3 and T4 levels remained normal in all groups. The occurrence of subclinical mastitis was lower in Day and Night groups than in Control and 24-hour groups. Regarding the microbiological analyses, in all groups the isolation of Corynebacterium sp. from milk samples increased while negative coagulase staphylococci (CNS) declined as etiological agents of subclinical mastitis. However, in Day and 24-hour groups, coagulase positive staphylococci (CPS) increased mainly Staphylococcus aureus (49.8% and 47.7% respectively). The Night group showed a decrease in subclinical mastitis occurrences. Our data indicate that all animals subjected to treatments presented high levels of cortisol, indicating a stress condition. The Night treatment presented a reduction in microbial isolation, suggesting a reduced susceptibility to mastitis.

Perfil energético e hormonal de ovelhas Santa Inês do terço médio da gestação ao pós-parto

No período periparto ocorrem importantes adequações fisiológicas que, se não forem efetivas predispõem a fêmea a enfermidades metabólicas. O conhecimento desta adaptação é relevante para que sejam implementadas, precocemente, medidas preventivas a poupar perdas produtivas. Com este objetivo foi avaliado o perfil energético e hormonal de ovelhas Santa Inês durante a gestação e puerpério. Foram utilizadas 10 ovelhas não gestantes (G0), 10 gestantes de um (G1) e 14 gestantes de dois e três fetos (G2). Foram avaliadas concentrações plasmáticas de glicose, ácidos graxos não esterificados (AGNE), betahidroxibutirato (BHB), e as concentrações séricas de insulina, glucagon, cortisol, triiodotironina (T3) e tiroxina (T4) a partir do 88º dia de gestação até o 28º dia pós-parto. No terço final de gestação, ovelhas gestantes apresentaram maiores concentrações de AGNE, T3 e T4 que as ovelhas não gestantes. No momento do parto foram observadas maiores concentrações de glicose, AGNE e T3 para todas as ovelhas gestantes em relação às não gestantes. Não houve diferença entre as ovelhas gestantes de um, dois ou três fetos. As diferenças observadas ocorreram apenas entre ovelhas gestantes e as vazias. Portanto, quando há adequada adaptação neste período de elevado desafio metabólico, os parâmetros bioquímicos aqui considerados independem do número de fetos gestados e podem ser considerados como valores de referência para ovelhas gestantes de um feto ou mais fetos do terço médio de gestação ao primeiro mês pós-parto.

Homogeneous assay technologies in drug screening: Quenching Resonance Energy Transfer (QRET) technique

Information gained from the human genome project and improvements in compound synthesizing have increased the number of both therapeutic targets and potential lead compounds. This has evolved a need for better screening techniques to have a capacity to screen number of compound libraries against increasing amount of targets. Radioactivity based assays have been traditionally used in drug screening but the fluorescence based assays have become more popular in high throughput screening (HTS) as they avoid safety and waste problems confronted with radioactivity. In comparison to conventional fluorescence more sensitive detection is obtained with time-resolved luminescence which has increased the popularity of time-resolved fluorescence resonance energy transfer (TR-FRET) based assays. To simplify the current TR-FRET based assay concept the luminometric homogeneous single-label utilizing assay technique, Quenching Resonance Energy Transfer (QRET), was developed. The technique utilizes soluble quencher to quench non-specifically the signal of unbound fraction of lanthanide labeled ligand. One labeling procedure and fewer manipulation steps in the assay concept are saving resources. The QRET technique is suitable for both biochemical and cell-based assays as indicated in four studies:1) ligand screening study of β2 -adrenergic receptor (cell-based), 2) activation study of Gs-/Gi-protein coupled receptors by measuring intracellular concentration of cyclic adenosine monophosphate (cell-based), 3) activation study of G-protein coupled receptors by observing the binding of guanosine-5’-triphosphate (cell membranes), and 4) activation study of small GTP binding protein Ras (biochemical). Signal-to-background ratios were between 2.4 to 10 and coefficient of variation varied from 0.5 to 17% indicating their suitability to HTS use.

Hormonal response during a fenfluramine-associated panic attack

Secretion curves for prolactin, cortisol, TSH, and GH from a 37-year old woman with dysthymia and panic disorder with agoraphobia were determined one day prior to (day I), and during a panic attack (day II) associated with an oral dose of 60 mg dl-fenfluramine, a drug known to increase anticipatory anxiety. The increased cortisol secretion observed is discussed in relation to the hormonal correlates of anxiety and the possible role of depression, dl-fenfluramine, and serotonergic receptor sensitivity

Percutaneous 17ß-estradiol replacement therapy in hypertensive postmenopausal women

The present study evaluated the short-term effects of percutaneous 17ß-estradiol on blood pressure, metabolic profile and hormonal levels in postmenopausal women with systemic arterial hypertension. After a wash-out period of 15 days, 10 hypertensive patients were treated with guanabenz acetate to control blood pressure, followed by 17ß-estradiol in the form of hydroalcoholic gel administered for 21 of 28 days of each cycle, for 3 cycles. Patients were evaluated before, during and 2 months after estrogen administration. Systolic and diastolic blood pressure or heart rate did not present any significant change in any patient when compared to those periods with the antihypertensive drug only (pretreatment period and 60 days after estrogen therapy was discontinued). Plasma biological markers of hepatic estrogenic action (plasma renin activity, antithrombin III, triglycerides, total cholesterol and lipoproteins) also remained unchanged during the study. Hormone treatment was effective, as indicated by the relief of menopausal symptoms, a decrease in FSH levels (73.48 ± 27.21 to 35.09 ± 20.44 IU/l, P<0.05), and an increase in estradiol levels (15.06 ± 8.76 to 78.7 ± 44.6 pg/ml, P<0.05). There was no effect on LH (18.0 ± 9.5 to 14.05 ± 8.28 IU/l). Hormone levels returned to previous values after estrogen treatment was discontinued. The data indicate that short-term percutaneous 17ß-estradiol replacement therapy, at the dose used, seems to be a safe hormone therapy for hypertensive menopausal women. Nevertheless, a controlled, prospective, randomized clinical assay with a larger number of subjects is needed to definitely establish both the beneficial and harmful effects of hormone replacement therapy in hypertensive women