We studied the levels and patterns of genetic variability in eight species of native fishes from the Valdivia River Basin (Región de los Rios, Chile), with the aim to estimate microevolutionary differences in species with similar ecological and historical conditions. The species considered were Diplomystes camposensis, Percilia gillissi, Galaxias platei, G. maculatus, Basilichthys australis, Percichthys trucha, Aplochiton zebra and A. taeniatus. We compared genetic diversity indexes both intra and inter-zones, and genetic structure, based on sequences of the control region for different sections along the river. In addition, we recovered relationships among haplotypes and their spatial distribution for each species, based on statistical parsimony networks. Estimators of past demographic behavior (D Tajima, Fs-Fu and mismatch analysis were also obtained. The results suggest that both patterns of genetic diversity in the river such as demographic responses are species specific. The most variable and strongest structured species in the river were G. maculatus and A. taniatus. P. trucha showed high structure values but its genetic variation was lower than the first two species. The less variable and less subdivided species were D. camposensis and P. gillissi. The remaining species showed levels of variability and structure intermediate between the two extreme patterns mentioned above. The differences among species are attributed to idiosyncratic biological attributes such as level of specificity in the use of aquatic habitat, population abundances, and historical migration patterns. In the case of G. maculatus, high levels of variability are attributed to the historical maintenance of high and stable effective population sizes, due to the ability to use both marine and freshwater habitats. Low levels of genetic diversity and indicators of historical reduction in effective population size, suggests that D. camposensis and P.gillissi were more sensitive and more exposed to population reduction. Results are discussed regarding ecological, biological, and historical factors.

Estudiamos los niveles y patrones de variabilidad genética de ocho especies de peces nativos de la cuenca del río San Pedro (cuenca del río Valdivia, Región de los Ríos, Chile), con el objetivo de evaluar diferencias microevolutivas en especies sometidas a similares condiciones ecológicas e históricas. Las especies consideradas fueron: Diplomystes camposensis, Percilia gillissi, Galaxias platei, G. maculatus, Basilichthys australis, Percichthys trucha, Aplochiton taeniatus y A. zebra. Para ello comparamos indicadores de diversidad genética tanto intra como inter-zonas, e indicadores de estructuración, basados en secuencias de la región de control, para diferentes tramos a lo largo del desarrollo del río. También recuperamos relaciones de parentesco entre haplotipos y su distribución espacial, para cada especie, a partir de redes construidas por parsimonia estadística. Además estimamos indicadores de comportamiento demográfico en el pasado (D de Tajima, Fs de Fu y distribución de frecuencias de diferencias pareadas). Los resultados sugieren que, tanto los patrones de diversidad genética en el río, como las respuestas demográficas, son especie específicos. Las especies más variables, conjuntamente con un alto grado de estructuración y bajo flujo génico en el río son G. maculatus y A. taeniatus. P. trucha mostró altos niveles de estructuración pero valores menores de variabilidad que el par de especies anterior. Las especies menos variables y menos subdivididas en la cuenca son Diplomystes camposensis y P. gillissi. Las demás especies mostraron niveles de variabilidad y estructuración intermedios entre los patrones extremos señalados para las especies anteriores. Las diferencias entre especies se atribuyen a atributos biológicos idiosincráticos tales como nivel de especialización en el uso de hábitat acuático, abundancias poblacionales históricas y comportamientos migratorios. En el caso de Galaxias maculatus, los altos niveles de variabilidad son atribuidos a la mantención histórica de altos tamaños poblacionales, gracias a la posibilidad de usar tanto hábitats marinos como dulceacuícolas. Mientras que los bajos niveles de diversidad e indicadores de variación histórica de tamaños efectivos poblacionales, sugieren que Diplomystes camposensis y Percilia gillissi fueron especies más sensibles y sometidas a reducciones poblacionales en el pasado. Se discuten los resultados en torno a factores ecológicos, biológicos e históricos.

RESUMO Foram avaliados soros de suínos provenientes de propriedades rurais que desenvolvem agricultura familiar no Município de Monte Negro, Rondônia, frente a agentes bacterianos e virais. Para a pesquisa de anticorpos contra Brucella spp., foram utilizados o Antígeno Acidificado Tamponado (AAT), a Soroaglutinação Lenta em Tubos (SAL) e a prova do 2-Mercaptoetanol (2-ME). Para a pesquisa de Leptospira spp. utilizou-se a Soroaglutinação Microscópica (SAM) contra 24 sorovares de leptospira. Anticorpos contra Parvovírus Suíno (PVS) foram pesquisados pela Inibição da Hemaglutinação (IH), enquanto que o vírus da Peste Suína Clássica (PSC) e da Doença de Aukeszky (DA) por Ensaio Imunoenzimático (ELISA). Apenas uma (0,9%) das 104 amostras testadas foi reagente na AAT e nenhuma na SAL e 2-ME. Para Leptospira spp. foram testados 88 soros, sendo detectados anticorpos em 29 amostras (32,9%) e os sorovares mais freqüentes foram Castellonis, Bratislava e Canicola. Foram detectados anticorpos contra PVS em 7,7% (8/104) dos soros testados. Nenhuma amostra foi reagente para PSC e DA.

ABSTRACT Sera from swine fromfamily-employed farms in Monte Negro County, state of Rondônia, Brazil, were evaluated againstbacterial and viral agents. Brucellosis was evaluated by Rose-Bengal agglutination (RBA), standard tube agglutination (STA), and mercaptoethanol (ME); and leptospirosis by microscopic agglutination test (MAT) against 24 leptospira serovars. Anti-porcine parvovirus antibodies were tested by haemmaglutination inhibition assay, and ELISA technique was used for detection of antibodies against classical swine fever virus and Aujeszky disease virus. Only 1 (0.9%) from 104 tested sera reacted by RBA, and no samples reacted in STA and ME. Leptospira spp. antibodies were detected in 29 (32.9%) of 88 tested sera and the most frequent serovars detected were Castellonis, Bratislava and Canicola. Antibodies against porcine parvovirus were detected in 7.7% (8/104) of the sera. None of the examined samples reacted to classical swine fever virus or Aujeszky disease virus.

Fifty-seven type 2 diabetic patients with metabolic syndrome and on insulin were assessed by a paired analysis before and 6 months after addition of metformin as combination therapy to evaluate the impact of the association on glycemic control, blood pressure, and lipid profile. This was a historical cohort study in which the files of type 2 diabetic patients with metabolic syndrome on insulin were reviewed. The body mass index (BMI), waist circumference, lipid profile, A1C level, fasting blood glucose level, daily dose of NPH insulin, systolic blood pressure, and diastolic blood pressure were assessed in each patient before the start of metformin and 6 months after the initiation of combination therapy. Glycemic control significantly improved (P < 0.001) after the addition of metformin (1404.4 ± 565.5 mg/day), with 14% of the 57 patients reaching A1C levels up to 7%, and 53% reaching values up to 8%. There was a statistically significant reduction (P < 0.05) of total cholesterol (229.0 ± 29.5 to 214.2 ± 25.0 mg/dL), BMI (30.7 ± 5.4 to 29.0 ± 4.0 kg/m²), waist circumference (124.6 ± 11.7 to 117.3 ± 9.3 cm), and daily necessity of insulin. The reduction of total cholesterol occurred independently of the reductions of A1C (9.65 ± 1.03 to 8.18 ± 1.01%) and BMI and the reduction of BMI and WC did not interfere with the improvement of A1C. In conclusion, our study showed the efficacy of the administration of metformin and insulin simultaneously without negative effects. No changes were detected in HDL-cholesterol or blood pressure.

Fetuses of mothers with gestational diabetes mellitus are at increased risk to develop perinatal complications mainly due to macrosomia. However, in view of the marked heterogeneity of this disease, it seems difficult to set guidelines for diagnosis and treatment. This complicates the choice of assigning patients either to diet or to insulin therapy. Also of concern is how much benefit could be expected from insulin therapy in preventing fetal complications in these patients. In a systematic review of the literature assessing the efficacy of insulin in preventing macrosomia in fetuses of mothers with gestational diabetes, we found six randomized controlled trials comparing diet alone to diet plus insulin. The studies included a total of 1281 patients (644 in the diet plus insulin group and 637 in the diet group), with marked differences among trials concerning diagnostic criteria, randomization process and treatment goals. Meta-analysis of the data resulted in a risk difference of -0.098 (95%CI: -0.168 to -0.028), and a number-necessary-to-treat of 11 (95%CI: 6 to 36), which means that it is necessary to treat 11 patients with insulin to prevent one case of macrosomia. This indicates a potential benefit of insulin, but not significantly enough to set treatment guidelines. Because of the heterogeneous evidence available in the literature about this matter, we conclude that larger trials addressing the efficacy of these two therapeutic modalities in preventing macrosomia are warranted.

Pancreatic ß cell function and insulin sensitivity, analyzed by the homeostasis model assessment, before and after 24 weeks of insulin therapy were studied and correlated with the presence of autoantibodies against ß cells (islet cell and anti-glutamic acid decarboxylase antibodies), in a group of 18 Brazilian lean adult non-insulin-dependent diabetes mellitus (NIDDM) patients with oral hypoglycemic agent failure (OHAF). Median fasting plasma glucose before and after insulin treatment was 19.1 and 8.5 mmol/l, respectively (P < 0.001); median HbA1c was 11.7% before vs 7.2% after insulin treatment (P < 0.001). Forty-four percent of the patients were positive (Ab+) to at least one autoantibody. Fasting C-peptide levels were lower in Ab+ than Ab- patients, both before (Ab+: 0.16 ± 0.09 vs Ab-: 0.41 ± 0.35 nmol/l, P < 0.003) and after insulin treatment (Ab+: 0.22 ± 0.13 vs Ab-: 0.44 ± 0.24 nmol/l, P < 0.03). Improvement of Hß was seen in Ab- (median before: 7.3 vs after insulin therapy: 33.4%, P = 0.003) but not in Ab+ patients (median before: 6.6 vs after insulin therapy: 20.9%). These results show that the OHAF observed in the 18 NIDDM patients studied was due mainly to two major causes: autoantibodies and ß cell desensitization. Autoantibodies against ß cells could account for 44% of OHAF, but Ab- patients may still present ß cell function recovery, mainly after a period of ß cell rest with insulin therapy. However, the effects of ß cell function recovery on the restoration of the response to oral hypoglycemic agents need to be determined.

The anthropometric status and metabolic control of 51 recently diagnosed Brazilian schoolchildren with type 1 diabetes (DM1), during the first 5 years of the disease, were compared with those of normal children (60 girls and 132 boys) belonging to the same environmental condition and pubertal stage. Metabolic control was evaluated on the basis of fasting plasma glucose (FPG) and HbA1c levels. The criteria of the National Center for Health Statistics were used for anthropometric evaluation. FPG (205 ± 51 mg/dl for girls vs 200 ± 34 mg/dl for boys) and % above upper normal limit of median HbA1c (1.8% for girls vs 2.5% for boys with diabetes) were not significantly different during follow-up. The Z-score of the last height evaluation was lower in the girls' group (-0.14 vs -0.53, P<0.05). By forward stepwise analysis, the Z-score of the initial height was statistically significant as a determinant factor for height at the end of the study in both girls and boys with DM1. The Z-score of weight at last evaluation was not different from that at diagnosis in either sex. However, analysis according to pubertal stage showed a tendency to a weight increase in the girls. The weight recovery and height loss in girls with DM1 follows the trend of the normal Brazilian population.

Adrenocortical autoantibodies (ACA), present in 60-80% of patients with idiopathic Addison's disease, are conventionally detected by indirect immunofluorescence (IIF) on frozen sections of adrenal glands. The large-scale use of IIF is limited in part by the need for a fluorescence microscope and the fact that histological sections cannot be stored for long periods of time. To circumvent these restrictions we developed a novel peroxidase-labelled protein A (PLPA) technique for the detection of ACA in patients with Addison's disease and compared the results with those obtained with the classical IIF assay. We studied serum samples from 90 healthy control subjects and 22 patients with Addison's disease, who had been clinically classified into two groups: idiopathic (N = 13) and granulomatous (N = 9). ACA-PLPA were detected in 10/22 (45%) patients: 9/13 (69%) with the idiopathic form and 1/9 (11%) with the granulomatous form, whereas ACA-IIF were detected in 11/22 patients (50%): 10/13 (77%) with the idiopathic form and 1/9 (11%) with the granulomatous form. Twelve of the 13 idiopathic addisonians (92%) were positive for either ACA-PLPA or ACA-IIF, but only 7 were positive by both methods. In contrast, none of 90 healthy subjects was found to be positive for ACA. Thus, our study shows that the PLPA-based technique is useful, has technical advantages over the IIF method (by not requiring the use of a fluorescence microscope and by permitting section storage for long periods of time). However, since it is only 60% concordant with the ACA-IIF method, it should be considered complementary instead of an alternative method to IIF for the detection of ACA in human sera.