Activation of C-C motif chemokine receptor 2 modulates testicular macrophages number, steroidogenesis, and spermatogenesis progression.

The monocyte chemoattractant protein 1 (MCP-1) belongs to the CC chemokine family and acts in the recruitment of C-C motif chemokine receptor 2 (CCR2)-positive immune cell types to inflammation sites. In testis, the MCP-1/CCR2 axis has been associated with the macrophage population's functional regulation, which presents significant functions supporting germ cell development. In this context, herein, we aimed to investigate the role of the chemokine receptor CCR2 in mice testicular environment and its impact on male sperm production. Using adult transgenic mice strain that had the CCR2 gene replaced by a red fluorescent protein gene, we showed a stage-dependent expression of CCR2 in type B spermatogonia and early primary spermatocytes. Several parameters related to sperm production were reduced in the absence of CCR2 protein, such as Sertoli cell efficiency, meiotic index, and overall yield of spermatogenesis. Daily sperm production decreased by almost 40%, and several damages in the seminiferous tubules were observed. Significant reduction in the expression of important genes related to the Sertoli cell function (Cnx43, Vim, Ocln, Spna2) and meiosis initiation (Stra8, Pcna, Prdm9, Msh5) occurred in comparison to controls. Also, the number of macrophages significantly decreased in the absence of CCR2 protein, along with a disturbance in Leydig cell steroidogenic activity. In summary, our results show that the non-activation of the MCP-1/CCR2 axis disturbs the testicular homeostasis, interfering in macrophage population, meiosis initiation, blood-testis barrier function, and androgen synthesis, leading to the malfunction of seminiferous tubules, decreased testosterone levels, defective sperm production, and lower fertility index.

Tadpoles fed supplemented diet with probiotic bacterium isolated from the intestinal tract of bullfrog Lithobates catesbeianus: Haematology, cell activity and electron microscopy.

The aim of this study is to select and isolate autochthonous bacteria with probiotic potential for use in a supplemented diet for bullfrog tadpoles, Lithobates catesbeianus. A total of 20 strains of lactic acid bacteria were isolated. Nine out of these were used in the following in vitro assays: antagonism against pathogenic bacteria (ANT), antimicrobial activity from extracellular compounds (MIC), tolerance to bile salts (TBS), pH reduction, protease production, sensitivity to antimicrobial tetracycline, cell viability, growth rate and doubling time. Using these data was defined an ideotype (ideal strain) based on the best results. Distances were estimated with the Mahalanobis (D2) test, and the best candidates, presenting the shortest ideotype distances, were considered to be used. The best strain was found to be Lactobacillus plantarum because it presented 10.00 ± 0.50 mm of ANT against Aeromonas hydrophila, 3.99 ± 0.01 of MIC independent of pathogenic bacteria, 85.07 ± 0.01 of TBS, 4.20 ± 0.02 of final pH, 17.67 ± 1.15 of protease production, 13.50 ± 2.00 sensitivity to antimicrobial tetracycline, 9.36 ± 0.04 of cell viability, 0.20 ± 0.00 of growth rate and 3.46 ± 0.00 doubling time. Therefore this probiotic candidate was then supplemented (2.045 ± 1.07 × 107 colony forming unities. g-1) into the diets of bullfrog tadpoles for a period of 42 days. At the end of the trial, samples of blood and intestines were collected to verify the haematological alterations and the intestinal morphology using transmission and scanning electron microscopy. Tadpoles fed the supplemented diet showed successful lactic acid bacterium colonisation, an increased number of circulating thrombocytes, monocytes, eosinophil and LG-PAS+ and also an increase in the length and density of intestinal microvilli. This study shows the feasibility of using probiotics isolated from farmed bullfrogs as a supplement in the diets of tadpoles, providing a promising alternative for modulating the health of these animals.

Visceral leishmaniasis after orthotopic liver transplantation: a rare cause of infection.

Visceral leishmaniasis (VL) was known as an opportunistic infection associated with immunosuppression, particularly related to human immunodeficiency virus infection and rarely to solid organ transplant recipients. We report a case of VL, 6 months after liver transplantation, in a patient who presented with febrile pancytopenia. The diagnosis was made by demonstration of amastigotes in smears from bone marrow. VL is a very rare infection in patients who undergo liver transplantation and, to our knowledge, this is the first case diagnosed in Portugal.

Biofilms from a Brazilian water distribution system include filamentous fungi.

Filamentous fungi in drinking water can block water pipes, can cause organoleptic biodeterioration, and are a source of pathogens. There are increasing reports of the involvement of the organisms in biofilms. This present study describes a sampling device that can be inserted directly into pipes within water distribution systems, allowing biofilm formation in situ. Calcofluor White M2R staining and fluorescent in situ hybridization with morphological analyses using epifluorescent microscopy were used to analyse biofilms for filamentous fungi, permitting direct observation of the fungi. DAPI (4',6-diamidino-2-phenylindole) was applied to detect bacteria. Filamentous fungi were detected in biofilms after 6 months on coupons exposed to raw water, decanted water and at the entrance of the water distribution system. Algae, yeast, and bacteria were also observed. The role of filamentous fungi requires further investigations.

Ghrelin improves burn-induced delayed gastrointestinal transit in rats.

Delayed gastrointestinal transit is common in patients with severe burn. Ghrelin is a potent prokinetic peptide. We aimed at testing the effect of ghrelin on burn-induced delayed gastrointestinal transit in rats. Gastric emptying (GE), intestinal transit (IT), and colonic transit (CT) studies were performed in male Sprague-Dawley rats. Rats were randomized into two main groups as follows: sham injury and ghrelin-treated burn injury with doses of 0, 2, 5, and 10 nmol/rat ip 6 h after burn. Sham/burn injury was induced under anesthesia. Rats received a phenol red meal 20 min following ghrelin injection. Based on the most effective ghrelin dose, 1 mg/kg sc atropine was given 30 min before the ghrelin in one group of rats for each study. The rats in each group were killed 30-90 min later; their stomachs, intestines, and colons were harvested immediately, and the amount of phenol red recovered was measured. Percentage of gastric emptying (GE%) and geometric center for IT and CT were calculated. We found 1) severe cutaneous burn injury significantly delayed GE, IT, and CT compared with sham injury (P < 0.05); 2) ghrelin normalized both GE and IT, but not the CT; 3) the most effective dose of ghrelin was 2 nmol/rat; and 4) atropine blocked the prokinetic effects of ghrelin on GE% and IT. In conclusion, ghrelin normalizes burn-induced delayed GE and IT but has no effect on CT in rats. The prokinetic effects of ghrelin are exerted via the cholinergic pathway. Ghrelin may have a therapeutic potential for burn patients with delayed upper gastrointestinal transit.

[Prognosis value of p53, C-erB-2 and Ki67 proteins in ovarian carcinoma]. / Valor prognóstico das proteínas p53, C-erB-2 E Ki67 no carcinoma do ovário.

The prognosis in ovarian carcinoma remains poor. We need to identify patients who are less likely to respond to treatment. In order to evaluate the prognostic value of C-erb-B2, p53 and Ki 67 expression and correlate these markers with classic prognostic factors, we studied paraffin-embedded tumor tissue from 81 patients with epithelial ovarian cancer and made a quantitative evaluation of C-erb-B2, p53 and Ki 67 expression by immunohistochemistry. The results were: age 5.4 +/- 15(22-88); 66% with normal physical activity; 48.2% with residual disease < 2 cm; initial stage--42% and advanced stage--58%. Age, performance status, residual disease and stage were correlated with 2 and 5 years survival. Positive immunostaining: p53--87%, C-erb B-2--51% and Ki67--100%. P53 and C-erb B-2 were associated with residual disease and stage; patients with no C-erbB-2 staining had a significantly better survival. A direct and significant correlation was found between p53 and Ki67 and between C-erb B-2 and p53. We conclude that these markers have a high expression in ovarian carcinoma and p53 and C-er B-2 correlate with stage and residual disease. Although C-erb B-2 was associated with better survival, it was not found to be an independent prognostic factor.

Transvaginal ultrasonography and hysteroscopy in postmenopausal bleeding: a prospective study.

OBJECTIVE: To determine the diagnostic value of transvaginal ultrasonography and hysteroscopy in patients with postmenopausal bleeding. MATERIAL AND METHODS: Between January 1, 1998 and June 30, 1999, 88 outpatient women with postmenopausal bleeding were enrolled in a prospective study. They underwent transvaginal ultrasonography and hysteroscopy, and were submitted to directed biopsy during hysteroscopy. Findings were classified as normal endometrium, suggestive of atrophy, focal abnormality (benign or suspicious), and diffuse thickness (benign or suspicious). Data was compared with the final diagnosis, established by histological examination, as atrophy, benign pathology, atypical hyperplasia and endometrial carcinoma. RESULTS: Among 88 women enrolled, 15 were excluded because hysteroscopy was impossible, and four had abandoned the study. The histological findings were scanty material in 12 (17.4%), atrophy in 24 (34.8%), cystic atrophy in one (1.4%), normal endometrium in five (7.2%), tuberculous endometritis in one (1.4%), polyps in 12 (17.4%), leiomyoma in one (1.4%), non-atypical hyperplasia in three (4.3%), atypical hyperplasia in one (1.4%) and endometrial carcinoma in nine cases (13.0%). For the assessment of endometrial carcinoma, ultrasonography revealed sensitivity 77.8%, specificity 93.3%, positive predictive value 63.6%, negative predictive value 96.6%; and hysteroscopy revealed sensitivity of 88.9%, specificity 98.3%, positive predictive value 88.9%, negative predictive value 98.3%. The combined use of both methods revealed sensitivity 100%, specificity 91.7%, positive predictive value 64.3%, negative predictive value 100%. CONCLUSIONS: Both imagiological methods were found to be useful screening tests for endometrial carcinoma. Hysteroscopy was a superior diagnostic procedure.

Bioequivalence of two lamivudine tablet formulations.

The present study describes the determination of the bioavailability of a new commercial tablet formulation of lamivudine (CAS 134678-17-4) compared with a reference formulation. The comparative bioequivalence of the test and a reference formulation (each 3 x 150 mg) was assessed in 24 healthy volunteers by means of a randomized two-way crossover design. Prior to the study both the test and reference formulations were examined for conformation to chromatographic purity and drug content. Each volunteer received the test (T) and the reference formulation (R) with a one-week drug-free interval between administrations. The plasma concentrations of T were monitored over a period of 12 h after drug administration using a sensitive HPLC method. Pharmacokinetic parameters for T were determined from plasma concentration-time data. Statistical tests were carried out at 90% confidence intervals using a parametric method (three-way ANOVA) for AUC and Cmax, and non-parametric method for Tmax. The present study showed that both formulations were bioequivalent for the geometric mean of AUC(0-12), AUC0-infinity), Cmax, and Tmax at the 90% confidence interval. The bioavailability of the test (%) was 96.7, 93.3, 99.7, 100.3, respectively. The T:R ratio was, in each case, well within the acceptable range of 100 +/- 20%.

[Thoracic endometriosis]. / Endometriose torácica.

Endometriosis was first described by Russel more than one-hundred years ago and still remains a clinical entity of difficult comprehension, with totally aberrant symptomatology, particularly in extra-genital situations. The present article describes a clinical case of recurring right-sided catamenial pneumothorax in a 45 year-old caucasian woman. In her last episode of right chest pain the X-ray film showed, besides the presence of a pneumothorax, a nodular image, later identified by CT scan as being a transdiaphragmatic hernia with hepatic content. The patient was then submitted to diagnostic and therapeutic thoracotomy and the diaphragmatic endometriosis was confirmed. Finally, a total hysterectomy with bilateral salpingo-ophorectomy was performed and the patient's condition remained uneventful with combined hormone therapy. The clinical presentation, pathogenesis, and therapeutic resolution of this nosological entity are briefly discussed.

[Effect of combined conjugated estrogen-medrogestone replacement therapy on lipid profiles, climacteric symptoms and the endometrium]. / Effets sur les lipides, les symptômes climatériques et l'endomètre d'une association d'estrogènes conjugués et de médrogestone dans le traitement hormonal substitutif.

OBJECTIVE: To assess the effects of continuous administration of conjugated estrogen combined with sequential administration of medrogestone on lipid profiles, climateric symptoms and endometrial tolerance. METHODS: This multicenter open study was conducted for one year to assess the effects of a hormone replacement therapy (HRT) regimen using Premarin (0.625 mg/day 28d/28) combined with medrogestone 5mg for 12 days (d17-d28 of each 28-day cycle) on lipid profiles, climateric symptoms and cycle control in 228 post menopausal women with an intact uterus. The subjects were recruited in 23 centers in 7 countries in Europe and Asia. Serum lipid/lipoprotein levels were determined at baseline and at cycles 3, 6, 13; endometrium biopsies were performed at screening then at cycle 13. Climateric symptoms and bleeding patterns were recorded by the patients from daily diaries cards collected at baseline and at visits during cycle 3, 6, 9, and 13. RESULTS: By cycle 3, the conjugated estrogen-medrogestone combination induced significant modifications of the lipid profile which were judged favorable. These modifications were maintained throughout treatment. All the baseline values were within normal limits. Mean variations compared with baseline values (expressed in mmol/l) after cycles 3, 6, and 13 were -0.46, -0.54, and -0.46 for total cholesterol (p<0.05), + 0.053, + 0.057, and + 0.078 for HDL-cholesterol (p<0.05) and -0.556, -0. 542, and -0.493 for LDL-cholesterol (p<0.001) respectively. VLDL-cholesterol levels were unchanged. Triglycerides increased significantly though moderately: + 0.12, + 0.15, and + 0.15 mmol/l at cycles 3, 6, and 13 respectively. Endometrial biopsies obtained at cycle 13 (n=195) did not reveal any endometrial hyperplasia. Withdrowal bleeding was predictable for a 6 to 7.4 day interval. The incidence of irregular bleeding varied from 7 to 33% and decreased progressively over the 13-cycle treatment. The incidence of amenorrhea increased from 14 to 52% over the 12 months studied. Finally, at each cycle, menopausal symptoms (mean number of hot flushes/day and Küpperman score) were significantly improved compared with the baseline. As expected, modifications were more pronounced after cycle 1, but improvements were maintained throughout the study. CONCLUSION: Continuous administration of Premarin in combination with sequential administration of medrogestone was found to be an effective treatment for menopausal symptoms. It was associated with favorable modifications of the lipid profile and was safe for the endometrium.

[Factors of recurrence of intraepithelial lesions of the uterine cervix]. / Factores de recorrência das lesões intraepiteliais do colo do útero.

MATERIAL AND METHODS: A retrospective study (1994-1996) of 282 cases of intraepithelial lesions treated in the Gynecology Services of Coimbra University Hospitals and subsequent follow-up (9-45 months). The following parameters were evaluated as recurrence factors: the patient's age, lesion degree, associated HPV infection, treatment type, safety margin in the ablative treatments and interval free of illness. RESULTS: Of the 282 cases, 72.4% (N = 204) corresponded to HSIL and 27.6% (N = 78) to LSIL. The patients' mean was 36.3 +/- 9.1 [19-67] years. The most commonly used treatment types were the LLETZ (large loop excision of the transformation zone) (76.9%) followed by CO2 LASER vaporization (16%) and finally cold-knife conization (7.1%). The safety margins were determined in 184 cases and, on average, were of 3.2 +/- 2.4 [1-10] mm. Forty-four recurrences occurred (15.6%) of which 75% corresponded to HSIL lesions (N = 33) and 25% (N = 11) to LSIL. From the total number of recurrences the association to HPV infection was found in 24 of the cases (54.5%) and 75% of these (N = 19) corresponded to HSIL. The average age in the recurrence group was 38.2 +/- 8.7 [21-53] years and 35.9 +/- 9.2 [19-67] years in the group without recurrences (p > 0.05). In the LSIL group treated with CO2 LASER (N = 32) the recurrence rate was 15.6% versus 13.3% in the group treated with LLETZ (N = 45). In the HSIL that were treated with CO2 LASER vaporization (N = 13) the recurrence rate was 23%; in those treated with LLETZ (N = 172) there were 16.2% recurrences and in the group submitted to cold-knife conization (N = 19) that rate was 10.5%. CONCLUSIONS: The total rate of recurrence was 15.6% with the greatest incidence of lesions associated to HPV (p > 0.05). Regarding the particular case of the HSIL, a great number of recurrences with the destructive treatments (CO2 LASER) was observed when compared to the ablative treatments (cold-knife conization or LLETZ).

[Wertheim-Meigs procedure. 10-year results]. / Operação de Wertheim-Meigs. Resultados de 10 anos.

Wertheim-Meigs hysterectomy is used in the treatment of cervical cancer, but since 1988 we have also used it to stage and treat endometrial carcinoma. To evaluate the morbidity of Wertheim Meigs hysterectomy as well as node invasion incidence and its correlation with other prognostic factors, the authors made a retrospective study of 112 patients, from October 1986 to March 1996. Among the 112 cases evaluated, 52% had cervical carcinoma and 48% had endometrial carcinoma; mean ages were 45 +/- 10 and 60 +/- 8 years (p < 0.005). FIGO stage distribution was: I-94.8 and 33.3%; II-5.2 and 51.9%; III-0 and 13%; IV-0 and 1.8%, for cervical and endometrial carcinoma. Hemorrhagic and traumatic accidents happened in 24.3% and 1.7% respectively. We had 15.5% early post operative complications and 1.7% late post operative ones. The mean duration of surgery and hospitalization was 163 +/- 29 minutes and 10 +/- 5 days. Pelvic lymphadenectomy was performed in 70% of patients and para-aortic nodes were also excised in the other 30%. The mean number of lymphatic nodes excised were 17 +/- 9. Pelvic node invasion was found in 9% and in 3.7% of para-aortic ones. When we studied endometrial cancer, we found a positive correlation between external myometrium invasion and lymphovascular invasion and positive node (p < 0.05). From the data available we may conclude that the complication rate of Wertheim Meigs is quite important, even though all but one complication were transitory. It may be possible to make a better selection of patients who need this surgical procedure by correct evaluation of risk factors.

Malnutrition as a prognostic factor in lymphoblastic leukaemia: a multivariate analysis.

One hundred and twenty eight Brazilian children with lymphoblastic leukaemia were intensively treated with a Berlin-Frankfurt-Munich based protocol. More children had a white cell count above 50 x 10(9)/l (31%) then observed in developed countries. After a median follow up of 31 months (11-58 months), the estimated probability of relapse free survival was 41% (7%) for the whole group. After adjustment in the Cox's multivariate model, malnutrition was the most significant adverse factor affecting duration of complete remission. Age above 8 years and high peripheral white cell count were also significant adverse factors. Among the nutritional indices, the height for age and weight for age z scores were both significant, whether the cut off points of z-2 or z = -1.28 were chosen to define malnutrition. A strong statistical association between the two indices was found; the contribution of height for age z score to the prediction of relapse free survival was more significant. Children with height for age z score < -2 had a relapse risk of 8.2 (95% confidence interval 3.1 to 21.9) relative to children with z score > -2. The results of this study suggest that socioeconomic and nutritional factors should be considered in the prognostic evaluation of children with leukaemia in developing countries.

Hemoglobinas anormais: perfil estatístico em doadores de sangue do Centro de Hematologia e Hemoterapia de Minas Gerais / Haemoglobinophaties in capable and uncapable blood samples donors from the Centro de Hematologia e Hemoterapia de Minas Gerais

As autoras apresentam um estudo sobre a frequência de hemoglobinopatias em amostras de sangue de doadores aptos e inaptos do Centro de Hematologia e Hemoterapia de Minas Gerais. Foram observados percentuais de 4,7 porcento e 6,7 para doadores aptos e inaptos em relaçäo à frequência de hemoglobinopatias. Devido aos possíveis prejuízos causados por transfusäo de sangue com hemoglobinopatias e levando em consideraçäo a significativa frequência desta alteraçäo, as autoras sugerem a intruduçäo dos principais métodos de diagnóstico nos serviços de Hemoterapia

Causas da cegueira binocular no Instituto Penido Burnier nos anos de 1940, 1960 e 1980

Os aa. estudaram as causas de cegueira binocular no Instituto Penido Burnier, Campinas, SP. nos anos de 1940, 1960 e 1980. Foram estudados 322 casos cujas etiologias puderam ser bem definidas. Avulta entre causas de cegueira, o glaucoma tanto no adulto quanto na criança. Merecem destaque também a retinpatia diabética no adulto e a fibroplasia retrolenticular na criança

Cateterismo percutaneo da veia subclavia em recem-nascidos e lactentes.Experiencia de 85 casos consecutivos. / Percutaneous subclavian vein catheterization in newborn infant and suckling. Experience of 85 cases

O cateterismo percutaneo da veia subclavia vem sendo por muitos anos restringido a adultos e criancas acima de um ano de vida, nao sendo bem estabelecidos os motivos que levam a este tipo de limitacao. Os autores estudam 85 cateterismos consecutivos da veia subclavia, realizados percutaneamente por via infraclavicular em 74 recem-nascidos e lactentes. As principais indicacoes para o cateterismo foram: a) acesso a veia central para monitorar pressao venosa indicacoes e administracao de antibioticos, drogas vasoativas etc., em pacientes criticos (56 casos); b) nutricao parenteral (9 casos); c) reanimacao cardiopulmonar (9 casos) e d) outros fins (11 casos). As principais complicacoes da insercao do cateter foram pneumotorax em dois pacientes e hidrotorax em um. Outras complicacoes consideradas "minor" (mau posicionamento do cateter puncao arterial. Sepse pelo cateter, definida como episodio de sepse para o qual nenhuma fonte anatomica de infeccao foi identificada, ocorreu em apenas dois casos, o que corresponde a uma incidencia de 2,4% em relacao ao total de cateterismos. O estudo revela assim uma incidencia de complicacoes mecanicas e infecciosas decorrentes do cateterismo percuteneo da veia subclavia em crianca de baixa idade igual aos indices dos estudos realizados em adultos. O procedimento e portanto seguro e de valor inestimavel em recem-nascidos e lactentes criticamente doentes