Minor sperm abnormalities in young male post-pubertal patients with juvenile dermatomyositis

The objective of the present study was to identify sperm abnormalities in young male patients with juvenile dermatomyositis (JDM). In 2005, 18 male JDM patients, diagnosed according to the criteria of Bohan and Peter, were followed at the Pediatric Rheumatology Unit and Rheumatology Division, of our Institution. Of the 18 males, 11 were pre-pubertal and 7 were post-pubertal. Two of 7 post-pubertal JDM male patients were excluded: one for orchidopexy for cryptorchidism and the other for testicular ectopia in the left testis. The remaining 5 post-pubertal JDM patients were prospectively evaluated on the basis of two semen analyses, according to the World Health Organization (WHO), urologic evaluation, testicular Doppler ultrasound hormone profile. The data of the JDM patients were compared with those of 5 age-matched healthy controls. The median age 18, was similar in JDM patients and controls. All JDM patients had teratozoospermia (abnormal sperm morphology), as did 4 (80%) of the controls. One of JDM patients had previous oligoasthenoteratozoospermia treated with intravenous cyclophosphamide with normalization of the number and concentration of the sperm after 5 years. All sperm parameters (sperm concentration, total sperm count and total motile sperm count by WHO, and sperm morphology by Kruger strict criteria), testicular volumes by Prader orchidometer and ultrasound, and hormones were similar in JDM patients compared with controls. The frequency of anti-sperm antibodies was similar in both groups. All JDM patients had minor sperm abnormalities in the head, midpiece, and/or tail of spermatozoids. Serial semen analyses in larger study populations are necessary to identify the extent and duration of sperm abnormalities in male patients with idiopathic inflammatory myopathies.

Puberty in male collared peccary (Pecari tajacu) determined by quantitative analysis of spermatogenic cells

Biological studies are necessary for the management of wildlife in captivity, and knowledge of reproduction is one of the important features for increasing production. The objective of the research was to determine the age at which male collared peccaries reach puberty. Testicular samples of 15 animals, aged 7 to 16 months, distributed into five groups (G1, G2, G3, G4 and G5) were used. The testes showed considerably increased weight, length and width (p < 0.05) from G1 to G3, whereas, from this group onward, the development of this organ was slower. There was positive correlation (p < 0.001) between the following testicular parameters: weight and length (r = 0.97), weight and width (r = 0.88), length and width (r = 0.92). Regarding the diameter of seminiferous tubules, an increase was observed (p < 0.05) from G1 to G4. The total number of spermatogenic cells increased significantly (p < 0.05) until G3 and then it stabilized. There was also positive correlation between testis weight and tubular diameter (r = 0.99, p < 0.001), and testis weight and spermatogenic cells (r = 0.98, p < 0.001). The number of Sertoli cells decreased significantly (p < 0.05) from G1, when they were undifferentiated as support cells, to G5, when they occurred together with the complete line of spermatic cells. The results demonstrate that the reproductive development of peccaries can be classified into the following stages: impuberty (G1, 7-8 months); pre-pubertal (G2, 9-10 months); pubertal (G3, 11-12 months); post-pubertal 1 (G4, 13-14 months); and post-pubertal 2 (G5, 15-16 months). Based on the histological analyses, puberty in the male collared peccary was determined to occur between 11 and 12 months of age.

Porcine follicular fluid concentration of free insulin-like growth factor-I collected from different diameter ovarian follicles

The study aimed to quantify the concentrations of free IGF-I in serum and fluid of ovarian follicles in pre-pubertal gilts and describe the ovarian morphology by measuring the size of the ovaries and counting the number of surface follicles. Ovaries (n=1,000) from pre-pubertal gilts were obtained immediately after slaughter. A total of 10 samplings were performed, with ovaries obtained from 50 females for each collection. The follicles situated on the surface of each ovary were classified as small (SFs, 2 to 5mm in diameter) or large (LFs 6 to 10mm in diameter) and the follicular fluid was obtained by follicle aspiration. The collection of serum samples was performed after the gilts exsanguination using sterile tubes. From the pool of serum and follicular fluid obtained from 50 females, the concentration of free IGF-I was determined in each sample using an enzyme immunoassay kit (ELISA). The description of ovarian morphometry was performed in 100 ovaries from randomly selected gilts. The larger and smaller lengths of ovaries were measured, and the total number of SFs and LFs present on the surface of each ovary were also counted. The IGF-I concentration was greater (P<0.05) in LFs (170.92±88.29 ng/mL) compared with SFs (67.39±49.90ng/mL) and serum (73.48±34.63ng/mL). The largest and smallest length of the ovaries was 26.0±3.0 and 19.0mm ±2.0mm, respectively. The number of SFs (70.86±25.76) was greater (P<0.01) than LFs (6.54±5.26). The study concluded that LFs present greater levels of IGF-I when compared with SFs and blood, which is related to increased activity of the LFs and its differentiation to ovulation. In addition, ovaries of pre-pubertal gilts have a higher number of SFs compared to LFs. Therefore, our study demonstrated unique data regarding the physiological concentration of free IGF-I in ovarian follicles, that can be used in future research to evaluate the addition of this hormone in the in vitro production media of porcine embryos with the goal to improve the technique efficiency.

Animal model for age- and sex-related genotoxicity of diethylstilbestrol

Environmental xenoestrogens pose a significant health risk for all living organisms. There is growing evidence concerning the different susceptibility to xenoestrogens of developing and adult organisms, but little is known about their genotoxicity in pre-pubertal mammals. In the present study, we developed an animal model to test the sex- and age-specific genotoxicity of the synthetic estrogen diethylstilbestrol (DES) on the reticulocytes of 3-week-old pre-pubertal and 12-week-old adult BALB/CJ mice using the in vivo micronucleus (MN) assay. DES was administered intraperitoneally at doses of 0.05, 0.5, and 5 µg/kg for 3 days and animals were sampled 48, 72 and 96 h, and 2 weeks after exposure. Five animals were analyzed for each dose, sex, and age group. After the DES dose of 0.05 µg/kg, pre-pubertal mice showed a significant increase in MN frequency (P < 0.001), while adults continued to show reference values (5.3 vs 1.0 MN/1000 reticulocytes). At doses of 0.5 and 5 µg/kg, MN frequency significantly increased in both age groups. In pre-pubertal male animals, MN frequency remained above reference values for 2 weeks after exposure. Our animal model for pre-pubertal genotoxicity assessment using the in vivo MN assay proved to be sensitive enough to distinguish age and sex differences in genome damage caused by DES. This synthetic estrogen was found to be more genotoxic in pre-pubertal mice, males in particular. Our results are relevant for future investigations and the preparation of legislation for drugs and environmentally emitted agents, which should incorporate specific age and gender susceptibility.

Health management practices and disease prevalence in dairy sheep systems in Argentina

A cross-sectional survey of 19 dairy sheep farms in Argentina was carried out with the purpose to know farm management, health practices, and occurrence and mortality of diseases. The survey comprised 40% of all sheep milking farms in Argentina. A questionnaire was conducted by way of personal interviews with sheep owners during farm visits. The proportions of farms reporting routine vaccination for clostridial diseases, contagious ecthyma, pneumonia and mineral and vitamin parenteral administration were 63%, 47.3%, 16.6% and 42.1% respectively. Regular treatment against lice was used in 37.5% of the farms, and 89.5% o the farmers treated against gastrointestinal nematodes (GIN). The mean number of GIN drenches per farm was 2.26±1.78 annually. In 68.4% of the milking flocks the California Mastitis Test was regularly done and 55.6% of the farmer managers had sampled their flocks once a year for Brucella ovis antibodies. During the pre-mating period respectively 68.4% and 50% of farmers clinically examined their rams and ewe for general health and teeth condition. The udders of ewes were frequently inspected at the start of each milking period. The most important parasite problems noticed were GIN (reported by 57.9% of farmers), lice (57.9%) and scabies (10.5%) and the most frequent infectious diseases were ecthyma (73.7%), pneumonia and other respiratory problems (57.9%), clinical mastitis (55.6%), clostridial diseases (36.9%) and foot lameness (35.2%). Photosensitivity (47.4%) and ruminal acidosis (42.1%) were reported as other frequent toxic or metabolic disorders. Owners mentioned that the mean lifespan or milk productive time per ewe was 4.5±1.4 years. Perinatal lamb mortality was 8.5% and the total flock mortality rates, above the first 24 h of life was 6.9%. The high rates of lamb mortality during the pre-weaning (10.3%) and post-weaning (5.9%) periods indicate that this problem, as well as the most prevalent diseases, should be the subject of further studies.

Intraocular pressure measurement in sheep using an applanation tonometer

The purpose of this study was to evaluate and establish the mean values of IOP in healthy adult sheep using an applanation tonometer. Information on age, sex, and breed was obtained for all animals included in this study. Twenty five healthy sheep (Ovis aries), of the same breed (Texel), male or female, with three years of age, received an ophthalmic examination in both eyes, including pupillary reflexes, Schirmer tear test, slit lamp biomicroscopy, and fluorescein staining. For all ophthalmic testing, animals were gently physically restrained, with no pressure in the jugular area and the eyelids were carefully open. IOP was measured by applanation tonometry (Tonopen XL). The same examiner performed the tonometry; measurements were taken three times for each eye, and their average was recorded as the IOP of the animal. Statistical analysis was performed using paired t-test and values of P < 0.05 were considered significant. The mean intraocular pressure in the whole group of 50 eyes was of 16.36 +/- 2.19 mm Hg. The mean (SD) IOP in the right eye was of 15.96 +/- 2.02 mm Hg, while the mean (SD) IOP in the left eye was of 16.76 +/- 2.32 mm Hg. Significant differences in IOP were not found between right and left eyes. The applanation tonometer was adequate for measuring the intraocular pressure in sheep. Reference data will assist in diagnosing testing for ophthalmic disease in sheep, as¹ well as promote further studies in this area.

Correlation between grain nutritional content and pasting properties of pre-gelatinized red rice flour

As with any variety of rice, red rice characteristics are subject to varietal differences, growing conditions, types of processing, and nutritional and rheological properties. This study determined the nutritional characteristics (centesimal composition and minerals) and paste viscosity properties of raw grains of four red rice genotypes (Tradicional MNAPB0405, MNACE0501 and MNACH0501) and the paste viscosity properties of pre-gelatinized flours obtained at different cooking times (20, 30 and 40 min). The main nutritional properties were correlated with the pasting properties of the pre-gelatinized flours. The samples showed differences in nutritional properties and paste viscosity. MNAPB0405 and MNACE0501 showed higher levels of fiber and fat and provided higher caloric energy than Tradicional and MNACH0501, which, in turn, showed higher levels of amylose. MNACH0501 showed higher peak viscosity (2402 cP), higher breakdown viscosity (696 cP) and a greater tendency to retrogradation (1510 cP), while Tradicional, MNAPB0405 and MNACE0501 had pasting profiles with peak viscosities varying between 855 and 1093 cP, breaking viscosity below 85 cP and retrogradation tendency between 376 and 1206 cP. The factors genotype and cooking time influenced the rheological behavior of pre-gelatinized flours, decreasing their pasting properties. The protein and amylose levels are correlated with the pasting properties and can be used as indicators of these properties in different genotypes of red rice, whether raw or processed into pre-gelatinized flours.

Effectiveness of see-and-treat for approaching pre-invasive lesions of uterine cervix

OBJECTIVE: To compare the effectiveness between the see-and-treat (S&T) approach and the conventional one (with prior biopsy) for squamous intraepithelial lesions of uterine cervix. METHODS: A cross-sectional study was conducted with 900 nonpregnant women with cytology suggestive of high grade squamous intraepithelial lesions in the city of Rio de Janeiro, Southeastern Brazil, between 1998 and 2004. The S&T approach consists of a large loop excision of the transformation zone procedure and is recommended when cytology is suggestive of high grade squamous intraepithelial lesion, satisfactory colposcopy with abnormalities compatible with the suspected cytological results, and the lesion is limited to the ectocervix or extends up to one centimeter of the endocervical canal. A subgroup of 336 patients whose colposcopy was considered satisfactory was analyzed, and they were divided into two groups for comparison: patients treated without prior biopsy (n = 288) and patients treated after a biopsy showing high grade squamous intraepithelial lesions (n = 48). Patients who were not treated or only treated more than a year later after recruitment at the colposcopy unit were considered dropouts. RESULTS: Of patients recruited during the study period, 71 were not treated or were only treated for at least a year. The overall dropout rate was 7.9% (95% CI: 6.1;9.7). Mean time elapsed between patient recruitment and treatment was 17.5 days in the S&T group and 102.5 days in the prior biopsy group. Dropout rates were 1.4% (95% CI: 0.04;2.7) and 5.% (95% CI: 0;12.3), respectively (p=0.07). The proportion of overtreated cases (negative histology) in the S&T group was 2.0% (95% CI: 0.4;3.6). CONCLUSIONS: The difference in the mean time elapsed between patient recruitment and treatment indicates that S&T is a time-saving approach The proportion of negative cases from using the S&T approach can be regarded as low.

Risk factors for osteoporotic fractures and low bone density in pre and postmenopausal women

OBJECTIVE: To estimate the prevalence and analyze risk factors associated to osteoporosis and low-trauma fracture in women. METHODS: Cross-sectional study including a total of 4,332 women older than 40 attending primary care services in the Greater São Paulo, Southeastern Brazil, between 2004 and 2007. Anthropometrical and gynecological data and information about lifestyle habits, previous fracture, medical history, food intake and physical activity were obtained through individual quantitative interviews. Low-trauma fracture was defined as that resulting from a fall from standing height or less in individuals 50 years or older. Multiple logistic regression models were designed having osteoporotic fracture and bone mineral density (BMD) as the dependent variables and all other parameters as the independent ones. The significance level was set at p<0.05. RESULTS: The prevalence of osteoporosis and osteoporotic fractures was 33% and 11.5%, respectively. The main risk factors associated with low bone mass were age (OR=1.07; 95% CI: 1.06;1.08), time since menopause (OR=2.16; 95% CI: 1.49;3.14), previous fracture (OR=2.62; 95% CI: 2.08;3.29) and current smoking (OR=1.45; 95% CI: 1.13;1.85). BMI (OR=0.88; 95% CI: 0.86;0.89), regular physical activity (OR=0.78; 95% CI: 0.65;0.94) and hormone replacement therapy (OR=0.43; 95% CI: 0.33;0.56) had a protective effect on bone mass. Risk factors significantly associated with osteoporotic fractures were age (OR=1.05; 95% CI: 1.04;1.06), time since menopause (OR=4.12; 95% CI: 1.79;9.48), familial history of hip fracture (OR=3.59; 95% CI: 2.88;4.47) and low BMD (OR=2.28; 95% CI: 1.85;2.82). CONCLUSIONS: Advanced age, menopause, low-trauma fracture and current smoking are major risk factors associated with low BMD and osteoporotic fracture. The clinical use of these parameters to identify women at higher risk for fractures might be a reasonable strategy to improve the management of osteoporosis.

Pre-harvest cane burning and health: the association between school absences and burning sugarcane fields

The objective of this study was to investigate an association between pre-harvest sugarcane burning and respiratory diseases in children under five years of age. The following data were collected in five schools in the city of Araraquara, SP, Southeastern Brazil, between March and June 2009: daily records of absences and the reasons stated for these absences, total concentration of suspended particulate matter (µg/m3), and air humidity. The relationship between the percentage of school absences due to respiratory problems and the concentration of particulate matter in March and from April to June presented a distinct behavior: absences increased alongside the increase in particulate matter concentration. The use of school absences as indicators of this relationship is an innovative approach.

Schistosoma mansoni circulating polysaccharide and protein antigens recognized by sheep antisera in patients with different clinical forms of schistosomiasis before and after treatment

Two sheep antisera, one of which raised against polysaccharide (Po) and other against protein (Pt) components of Schistosoma mansoni adult worms, were assessed by ELISA for their ability to detect circulating parasite antigens in patients with different clinical forms of chronic schistosomiasis mansoni. The former antiserum detected parasite antigens in liver granulomata and the latter in renal glomeruli from schistosomiasis patients and mice experimentally infected with S. mansoni. In general, the levels and/or positivity rate of circulating antigens and specific IgG antibodies were significantly higher in patients with hepatointestinal (HI) and hepatosplenic (HS) forms than in mild intestinal (I) forms. An association between Po antigens and clinical features of the disease was observed, as the level of these antigens was low (137 ng/ml) as well as the positivity rate (7.9%) in patients with I forms; values that were intermediate (593 ng/ml and 33.3%) in those with HI forms, and high (1.563 ng/ml and 50.0%) in more severe HS forms. The Pt antigens were detected in the studied clinical forms not differing statistically but, the positivity rate was significantly higher in HS forms comparatively to I forms. The antisera studied revealed distinct circulating antigen profiles, and the prognostic value of Po and Pt antigens was suggested.

Studies on human anti-rabies immunization in Brazil: I - Evaluation of the 3 + 1 pre-exposure vaccination schedule under field conditions

The currently used pre-exposure anti-rabies immunization schedule in Brazil is the one called 3+1, employing suckling mouse brain vaccine (3 doses on alternate days and the last one on day 30). Although satisfactory results were obtained in well controlled experimental groups using this immunization schedule, in our routine practice, VNA levels lower than 0.5 IU/ml are frequently found. We studied the pre-exposure 3+1 schedule under field conditions in different cities on the State of São Paulo, Brazil, under variable and sometimes adverse circumstances, such as the use of different batches of vaccine with different titers, delivered, stored and administered under local conditions. Fifty out of 256 serum samples (19.5%) showed VNA titers lower than 0.5 IU/ml, but they were not distributed homogeneously among the localities studied. While in some cities the results were completely satisfactory, in others almost 40% did not attain the minimum VNA titer required. The results presented here, considered separately, question our currently used procedures for human pre-exposure anti-rabies immunization. The reasons determining this situation are discussed.

Studies on human anti-rabies immunization in Brazil: II - Preliminary evaluation of the 2-1-1 schedule for human pre-exposure anti-rabies immunization, employing suckling mouse brain vaccine

This study reports preliminary results of virus neutralizing antibody (VNA) titers obtained on different days in the course of human anti-rabies immunization with the 2-1-1 schedule (one dose is given in the right arm and one dose in the left arm at day 0, and one dose is apllied on days 7 and 21), recommended by WHO for post-exposure treatment with cell culture vaccines. A variant schedule (double dose on day zero and another on day 14) was also tested, both employing suckling mouse brain vaccine. A complete seroconversion rate was obtained after only 3 vaccine doses, and almost all patients (11 of 12) presented titers higher than 1.0 IU/ml. Both neutralizing response and seroconversion rates were lower in the group receiving only 3 doses, regardless of the sample collecting day. Although our results are lower than those found with cell culture vaccines, the geometry mean of VNA is fully satisfactory, overcoming the lower limit recommended by WHO of 0.5 IU/ml. The 2-1-1 schedule could be an alternative one for pre exposure immunization, shorter than the classical 3+1 regimen (one dose on days 0, 2, 4 and 30) with only three visits to the doctor, instead of four.

Antigenic typing of brazilian rabies virus samples isolated from animals and humans, 1989-2000

Animal and human rabies samples isolated between 1989 and 2000 were typified by means of a monoclonal antibody panel against the viral nucleoprotein. The panel had been previously established to study the molecular epidemiology of rabies virus in the Americas. Samples were isolated in the Diagnostic Laboratory of the Pasteur Institute and in other rabies diagnostic centers in Brazil. In addition to the fixed virus samples CVS-31/96-IP, preserved in mouse brain, and PV-BHK/97, preserved in cell culture, a total of 330 rabies virus samples were isolated from dogs, cats, cattle, horses, bats, sheep, goat, swine, foxes, marmosets, coati and humans. Six antigenic variants that were compatible with the pre-established monoclonal antibodies panel were defined: numbers 2 (dog), 3 (Desmodus rotundus), 4 (Tadarida brasiliensis), 5 (vampire bat from Venezuela), 6 (Lasiurus cinereus) and Lab (reacted to all used antibodies). Six unknown profiles, not compatible with the panel, were also found. Samples isolated from insectivore bats showed the greatest variability and the most commonly isolated variant was variant-3 (Desmodus rotundus). These findings may be related to the existence of multiple independent transmission cycles, involving different bat species.