Neonatal inhalatory anesthetic exposure: Reproductive changes in male rats

We investigated the effects of an inhalatory anesthetic (ethyl ether) during the neonatal period of brain sexual differentiation on the later fertility and sexual behavior of male rats. Animals were exposed to ethyl ether immediately after birth. At adulthood, body weight, testes wet weight, and plasma testosterone levels were not affected; however, neonatal exposure to ether showed alterations on male fertility: a decrease in the number of spermatids and spermatozoa, an increase in the transit time of cauda epididymal spermatozoa and a decrease in daily sperm production. An alteration of sexual behavior was also observed: decreased male sexual behavior and appearance of homosexual behavior when the male rats were castrated and pretreated with exogenous estrogen. Probably, the ether delayed or reduced the testosterone peak of the sexual differentiation period, altering the processes of masculinization and defeminization of the hypothalamus. Our results indicate that perinatal exposure to ethyl ether during the critical period of male brain sexual differentiation, acting as endocrine disruptors, has a long-term effect on the fertility and sexual behavior of male rats, suggesting endocrine disruption through incomplete masculinization and defeminization of the central nervous system. © 2002 Elsevier B.V. All rights reserved.

Estudo do extrato de Ginkgo biloba na prevenção de malformações em recémnascidos de ratas diabéticas

The pregnant women presenting Diabetes mellitus develop metabolic alterations, that may cause damage to the fetal well-being and provoke anomalies and/or malformations. The antioxidant treatment has improved the embryonic development from streptozotocin diabetic rats. Several studies have shown that a Gingko biloba extract presents antioxidant effects and, in the present study, one of the G. biloba extract formulations was used (EGb761) - Tebonin (200 mg/Kg/day), given to the diabetic pregnant female rats. The aim was to evaluate the effect of the EGb761 treatment on the of anomalies and/ or malformations incidence of the offspring. Diabetes was induced in female rats using streptozotocin in a dose of 40 mg/kg. The rats were mated, and the pregnant animals were divided in two groups: Control (water) and experimental (G. biloba). At day 21 of pregnancy, the rats were killed, and their fetuses were analyzed and processed for anomalies and/or malformations incidence. The results demonstrated that control and experimental groups presented no external anomalies and malformations; increased incidence of skeletal anomalies and of visceral malformations, and lower rate of visceral anomalies and skeletal malformations. These data confirm no statistical difference and, therefore, EGb761 treatment did not cause changes. Thus, a dose of 200 mg/Kg/day of a Gingko biloba extract given during the pregnancy rat was ineffective in the prevention of the anomalies and/or malformations related to the diabetes.

Avaliação da fertilidade e de possíveis defeitos estruturais externos sobre a prole de ratos machos expostos cronicamente ao arsênio

Arsenic is an environmental pollutant that influences the male reproductive function, affecting the spermatogenesis process in the testis and causing alterations in the sperm. The objective of this work was evaluate the reproductive efficiency in adult male rats exposed to arsenic and the possible male-mediated structural malformations. The animal were treated with sodium arsenato (3,6 mg/Kg, i.p.), twice a weak, for 15, 30, 45, 60 or 75 days whereas control received only distilled water. Results have showed a reduced pregnancy rate associated to a decrease in the fertility potencial and to a increase in the preimplantation loss. There was a decrease in the fetal weights, as showed by its average. Malformations rate, postimplantation loss and resorptions were unaffect by treatment. These results suggest that chronic exposure to arsenic provokes reduction on fertility of male rats. Further studies are required to clarify the arsenic effects on male reproductive system and male-mediated effects on progeny.

Mixture of vitamin C, hesperidin and piperidol exposure in pregnancy: Maternal-fetal repercussions

To evaluate the reproductive performance and the development of their offspring on rat pregnancy. Wistar pregnant rats were gavaged with 0 mg/kg wb/day (control group, n = 20) and 166.5 mg/kg/day of a mixture of vitamin C, hesperidin and piperidol (experimental group, n = 20) during the organogenic period (from day 5 to 14 of pregnancy; positive vaginal smear = day 0). The female rats were killed on day 21 of pregnancy. The number of implantations, resorptions (dead embryos), and live/dead fetuses were counted for the analysis of the postimplantation loss rates. There was neither alteration in maternal reproductive performance, but it was verified an increase of the number of fetuses presenting dilated urether, hydronephrosis, and reduced ossification of skull due to the treatment of female rats with a mixture of vitamin C, hesperidin and piperidol, these abnormalities were considered transitory and may not interfere on offspring development. It was not verified other type of major malformation neither the appearance of fetuses presenting atrophy of upper limbs that it could be associated to use of this drug.

Determinação precoce do sexo fetal pela análise do DNA no plasma materno

Purpose: to verify the viability of early diagnosis of fetal gender in maternal plasma by the real-time polymerase chain reaction (real-time PCR) starting at the 5th week of pregnancy. Methods: peripheral blood was collected from pregnant women with single fetus starting at the 5th week of gestation. After centrifugation, 0.4 mL plasma was separated for fetal DNA extraction. The DNA was analyzed in duplicate by real-time PCR for two genomic regions, one of the Y chromosome and the other common to both sexes, through the TaqMan® method, which uses a pair of primers and a fluorescent probe. Patients who aborted were excluded. Results: a total of 79 determinations of fetal DNA in maternal plasma were performed in 52 pregnant women. The results of the determinations were compared to fetal gender after delivery. Accuracy according to gestational age was 92.6% (25 of 27 cases) at 5 weeks with 87% sensitivity, and 95.6% (22 of 23 cases) at 6 weeks with 92% sensitivity. Starting at the 7th week of pregnancy, accuracy was 100% (29 of 29 cases). Specificity was 100% regardless of gestational age. Conclusion: real-time PCR for the detection of fetal gender in maternal plasma starting at the 5th week of gestation has good sensitivity and excellent specificity. There was agreement of the results in 100% of the cases in which male gender was diagnosed, regardless of gestational age, and from the 7th week of gestation for female gender diagnosis.

Aspectos estruturais da membrana eritrocitária

This article describes the structures and functions of the erythrocyte membrane and its importance in transfusional medicine. The erythrocyte membrane is one of the best known membranes in terms of structure, function and genetic disorders. As any other plasma membrane, it mediates transport functions. It also provides the erythrocytes with their resilience and deformability. According to the International Society of Blood Transfusion (ISBT), more than 500 antigens are expressed in the erythrocyte membrane, and around 270 are involved in transfusion reaction cases and hemolytic diseases of the fetus and newborn. In the ISBT classification, the high frequency series is represented by antigens in more than 99% of population (high prevalence antigen). In transfusion, the absence of these antigens determines severe problems as for example, one woman without the P antigen suffered 6 repetitive miscarriages due to placental insufficiency, which was caused by an antibody formed against the absent P antigen. Some important erythrocyte membrane proteins are described here including Band 3, Glycophorins and spectrin. The most abundant integral membrane protein is Band 3 and its main function is to mediate exchange of chloride and bicarbonate anions across the plasma membrane. The second most abundant integral membrane protein in the human erythrocyte is sialoglycoprotein glycophorin A (GPA). With its high sialic acid content, GPA is the main contributor to the net negative cell-surface charge and is thus critical for minimizing cell-cell interactions and preventing red cell aggregation. Glycophorin C (GPC) is the receptor for PfEBP-2 (baebl, EBA-140), the newly identified erythrocyte binding ligand of Plasmodium falciparum. The ternary complex of spectrin, actin and 4.1R defines the nodes of the erythrocyte membrane skeletal network, and is inseparable from membrane stability when under mechanical stress. This erythrocyte membrane review is important for a better understanding of transfusion reactions, where the antibody formation against high prevalence antigens makes compatible transfusions difficult. The study of antigen diversity and biochemical characterization of different proteins will contribute to healthcare, as well as diagnosis, development of technology such as monoclonal antibody production and the therapeutic conduct of many diseases.

The materno-fetal interface in llama (Lama guanicoe glama)

Samples from 9 llamas (28 through 36 weeks of gestation) were collected and fixed in 4% buffered paraformaldehyde (light microscopy) and in 2.5% buffered glutaraldehyde (transmission and scanning electron microscopy). The material was processed in paraplast and slides (5μm) were stained with HE, PAS, Masson-Trichrome, acid phosphatase and Perl's. The uteroferrin was immunolocalized. The results show that llama placenta is chorioallantoic, diffuse, folded and epitheliochorial, and the fetus is covered with an epidermal membrane. The trophoblast cells have variable morphology: cubic, rounded and triangular cells, with cytoplasm containing PAS-positive granules. Binucleated cells with large cytoplasm and rounded nuclei, as well as giant trophoblastic cells with multiple nuclei were also observed. Numerous blood vessels were observed beneath the cells of the uterine epithelium and around the chorionic subdivided branches. Glandular activity was shown by PAS, Perl's, and acid phosphatase positive reactions in the cytoplasm and glandular lumen, and by immunolocalization of the uteroferrin in the glandular epithelium. The uterine glands open in spaces formed by the areoles, which are filled by PAS-positive material. The llama fetus was covered by the epidermal membrane, composed of stratified epithelium, with up to seven layers of mono-, bi- or trinucleated cells. The high level of maternal and fetal vascularization surfaces indicates an intense exchange of substances across both surfaces. The metabolic activity shown in the uterine glands suggests an adaptation of the gestation to the high altitudes of the natural habitat of this species.

Susceptibilidade in vitro a antimicrobianos da Mannheimia haemolytica e da Pasteurella multocida isoladas de ovinos sadios e com doenças respiratórias

Pasteurella multocida and Mannheimia haemolytica (P. haemolytica) are associated with ovine respiratory diseases. With the purpose of observe the susceptibility in vitro of these organisms against antimicrobials, were collected samples of nasopharingeal (n=180) and oropharingeal (n=82) from ovines healthy and with respiratory disease. Among the antimicrobials tested, the sensibility was greater for enrofloxacin (100%) and florfenicol (100%), for both bacteria. The greater resistance indices for M. haemolytica and P. multocida were observed with tetracyclin (15.64% and 17.65% respectively) and penicillin (1.82% and 4.2%).

Síndrome de Eisenmenger na gravidez

Eisenmenger's syndrome consists of pulmonary hypertension with a reversed or bidirectional shunt at the atrioventricular, or aortopulmonary level. Eisenmerger's syndrome in pregnancy is usually associated with high mortality rates (nearly 30-50%). Unfortunately, pulmonary hypertension is aggravated during pregnancy and often leads to an unfavorable outcome. Here, we report a successful pregnancy in a woman with Eisenmenger syndrome.

Mecanismo de centralização: Da insuficiência placentária à adaptação circulatória fetal

The application and development of obstetric Dopplervelocimetry provide a basis for the investigation of placental insuf ciency and demonstrate the dynamic behavior of fetal circulation during hypoxia. In clinical practice, assessing hemodynamics in three vascular regions involved in pregnancy, namely the uterine, umbilical and middle cerebral arteries, has become routine. Roughly, the cerebral artery expresses the balance between uterine artery oxygen supply and umbilical artery oxygen uptake. Currently, when such balance is unfavorable, the fetal cardiac reserve is investigated by assessing the venous duct. However, determining and interpreting vascular resistance indexes is not an easy task. The starting point is to know the physiopathology of placental insuf ciency and fetal circulatory adaptation through which Doppler con rmed its role in the assessment of fetal well-being.

Influence of lower cutoff values for 100-g oral glucose tolerance test and glycemic profile for identification of pregnant women at excessive fetal growth risk

Objective: To evaluate data from patients with normal oral glucose tolerance test (OGTT) results and a normal or impaired glycemic profile (GP) to determine whether lower cutoff values for the OGTT and GP (alone or combined) could identify pregnant women at risk for excessive fetal growth. Methods: We classified 701 pregnant women with positive screening for gestational diabetes mellitus (GDM) into 2 categories - (1) normal 100-g OGTT and normal GP and (2) normal 100-g OGTT and impaired GP - to evaluate the influence of lower cutoff points in a 100-g OGTT and GP (alone or in combination) for identification of pregnant women at excessive fetal growth risk. The OGTT is considered impaired if 2 or more values are above the normal range, and the GP is impaired if the fasting glucose level or at least 1 postprandial glucose value is above the normal range. To establish the criteria for the OGTT (for fasting and 1, 2, and 3 hours after an oral glucose load, respectively), we considered the mean (75 mg/dL, 120 mg/dL, 113 mg/dL, and 97 mg/dL), mean plus 1 SD (85 mg/dL, 151 mg/dL, 133 mg/dL, and 118 mg/dL), and mean plus 2 SD (95 mg/dL, 182 mg/dL, 153 mg/dL, and 139 mg/dL); and for the GP, we considered the mean and mean plus 1 SD (78 mg/dL and 92 mg/dL for fasting glucose levels and 90 mg/dL and 130 mg/dL for 1- or 2-hour postprandial glucose levels, respectively). Results: Subsequently, the women were reclassified according to the new cutoff points for both tests (OGTT and GP). Consideration of values, in isolation or combination, yielded 6 new diagnostic criteria. Excessive fetal growth was the response variable for analysis of the new cutoff points. Odds ratios and their respective confidence intervals were estimated, as were the sensitivity and specificity related to diagnosis of excessive fetal growth for each criterion. The new cutoff points for the tests, when used independently rather than collectively, did not help to predict excessive fetal growth in the presence of mild hyperglycemia. Conclusion: Decreasing the cutoff point for the 100-g OGTT (for fasting and 1, 2, and 3 hours) to the mean (75 mg/dL, 120 mg/dL, 113 mg/dL, and 97 mg/dL) in association with the GP (mean or mean plus 1 SD-78 mg/dL and 92 mg/dL for the fasting state and 90 mg/dL and 130 mg/dL for 1- or 2-hour postprandial values-increased the sensitivity and specificity, and both criteria had statistically significant predictive power for detection of excessive fetal growth. © 2008 AACE.

Melatonin effects on macrophage in diabetic rats and the maternal hyperglycemic implications for newborn rats

The modulatory effects of melatonin (MLT) on maternal and fetal macrophages in diabetic rats and the repercussion of maternal hyperglycemia on fetus-placenta parameters were studied. This was achieved by determining maternal and fetal blood glucose, weight and superoxide release by macrophages. Placental weight, protein, DNA and RNA concentration were also verified. Superoxide levels in macrophages isolated from pregnant healthy rats were higher than those obtained from diabetic animals. Melatonin increased significantly in the macrophages of control animals (18.7 ± 2.8 with MLT compared to 14.2 ± 1.6 without MLT) but decreased with melatonin stimulation in diabetic rats (8.8 ± 1.4 with MLT compared to 12.9 ± 2.1 without MLT). Melatonin significantly decreased superoxide levels in newborns of diabetic mothers (7.3 ± 3.4) compared to those of healthy (14.6 ± 3.5) mothers. Blood glucose levels were significantly higher (p<0.05) in newborn rats of diabetic mothers (108.3 ± 7.8) compared to blood glucose levels in newborn control rats (81.2 ± 10.7). Body weight was significantly higher (p <0.05) in the offspring of rats with alloxan-induced diabetes. No statistical difference (p> 0.05) was observed in the placenta weight, total protein concentration and DNA of rats. The RNA concentration was significantly lower (p <0.05) in the placentas of rats with alloxan-induced diabetes (156.1 ± 71.8), when compared to the concentration of RNA in the placentas of control rats (239.5 ± 77.3). In conclusion, maternal hyperglycemia modified the fetus-placental parameters and melatonin modulated the macrophages activation in maternal and fetal diabetic rats.

Resposta imune e capacidade de neutralização de anticorpos produzidos em ovinos jovens imunizados com veneno de Crotalus durissus terrificus nativo e irradiado com Cobalto 60

The ELISA technique was used to evaluate and compare young ovine humoral immune response during crotalic antiserum production. Animals were clinically evaluated throughout this process, and the neutralizing capacity of antisera raised against natural (NV) and Cobalt-60 irradiated (IrV) venoms of Crotalus durissus terrificus (C.d.t.) was verified by means of in vitro challenges. Three groups of six animals each were used: G1 received NV; G2 was inoculated with IrV; and G3 was used as control. Animals received six immunizations during 84 days at 14-day intervals. ELISA of antibody profile showed significant difference (p<5%) between experimental groups (G1ovines to produce commercial crotalic antiserum, which may be employed in the treatment of human and animal envenomation. Production cost might be reduced by the subsequent utilization of hyperimmunized ovines as food.

Integrated analysis of halogenated organic pollutants in sub-millilitre volumes of venous and umbilical cord blood sera

A rapid, robust and economical method for the analysis of persistent halogenated organic compounds in small volumes of human serum and umbilical cord blood is described. The pollutants studied cover a broad range of molecules of contemporary epidemiological and legislative concern, including polychlorobiphenyls (PCBs), polychlorobenzenes (CBs), hexachlorocyclohexanes (HCHs), DDTs, polychlorostyrenes (PCSs) and polybromodiphenyl ethers (PBDEs). Extraction and clean-up with n-hexane and concentrated sulphuric acid was followed with analysis by gas chromatography coupled to electron capture (GC-ECD) and GC coupled to negative ion chemical ionisation mass spectrometry (GC-NICI-MS). The advantages of this method rest in the broad range of analytes and its simplicity and robustness, while the use of concentrated sulphuric acid extraction/clean-up destroys viruses that may be present in the samples. Small volumes of reference serum between 50 and 1000 μL were extracted and the limits of detection/quantification and repeatability were determined. Recoveries of spiked compounds for the extraction of small volumes (≥300 μL) of the spiked reference serum were between 90% and 120%. The coefficients of variation of repeatability ranged from 0.1-14%, depending on the compound. Samples of 4-year-old serum and umbilical cord blood (n = 73 and 40, respectively) from a population inhabiting a village near a chloro-alkali plant were screened for the above-mentioned halogenated pollutants using this method and the results are briefly described. © 2010 Springer-Verlag.