Studies of esterase 6 in Drosophila melanogaster. XVIII. Biochemical differences between the slow and fast allozymes

Most natural populations of Drosophila melanogaster are polymorphic for two major electrophoretic variants at the esterase-6 locus. The frequency of the EST 6F allozyme is greatest in populations in warmer latitudes, whereas the EST 6S allozyme is predominant in colder latitudes. Latitudinal clines in electromorph frequencies are found on three continents. Purified preparations of the allozymes have been characterized for their pH optimum, substrate specificity, organophosphate inhibition, alcohol activation, thermal stability, and kinetic parameters. These and previous analyses of the EST 6 allozymes reveal that the two variants have differences in their physical and kinetic properties that may provide a basis for the selective maintenance of the polymorphisms and an explanation of the clinal variation observed in natural populations.      

Sweat gland density and response during high-intensity exercise in athletes with spinal cord injuries

Sweat production is crucial for thermoregulation. However, sweating can be problematic for individuals with spinal cord injuries (SCI), as they display a blunting of sudomotor and vasomotor responses below the level of the injury. Sweat gland density and eccrine gland metabolism in SCI are not well understood. Consequently, this study examined sweat lactate (S-LA) (reflective of sweat gland metabolism), active sweat gland density (SGD), and sweat output per gland (S/G) in 7 SCI athletes and 8 able-bodied (AB) controls matched for arm ergometry VO₂peak. A sweat collection device was positioned on the upper scapular and medial calf of each subject just prior to the beginning of the trial, with iodine sweat gland density patches positioned on the upper scapular and medial calf. Participants were tested on a ramp protocol (7 min per stage, 20 W increase per stage) in a common exercise environment (21±1°C, 45-65% relative humidity). An independent t-test revealed lower (p<0.05) SGD (upper scapular) for SCI (22.3 ±14.8 glands · cm⁻²) vs. AB. (41.0 ± 8.1 glands · cm⁻²). However, there was no significant difference for S/G between groups. S-LA was significantly greater (p<0.05) during the second exercise stage for SCI (11.5±10.9 mmol · l⁻¹) vs. AB (26.8±11.07 mmol · l⁻¹). These findings suggest that SCI athletes had less active sweat glands compared to the AB group, but the sweat response was similar (SLA, S/G) between AB and SCI athletes. The results suggest similar interglandular metabolic activity irrespective of overall sweat rate.     

Clinical,ultrasonographic, and endocrinological studies on donkey pregnancy

Although donkey breeding has gained new interest in the past two decades, knowledge about donkey reproduction is still scarce, particularly on jenny pregnancy. The aim of this study was to describe the ultrasonographic and endocrine profiles of the physiological pregnancy in the jenny. The study was performed on 12 pregnancies of 7 Amiata donkeys from Day 10 after ovulation to delivery. Because three pregnancies, respectively at weeks 42, 44, and 45, were considered pathologic and treated pharmacologically, data collected from 2 weeks before diagnosis to the end of pregnancy were removed from the analysis. Average length of the normal pregnancies was 353.4 ± 13.0 days (range, 339–370 days). Timing, dimensions, and development during the first phases of embryonic growth, evaluated using transrectal ultrasound, were similar to that previously described in jennies and mares: first detection of embryonic vesicle was at 11.8 ± 1.3 days of gestation and diameter was 6.5 ± 1.9 mm, loss of spherical shape occurred at 18.5 ± 1.4 days, and embryo and heart beat were first seen at 22.0 ± 1.1 and 25 ± 1.1 days, respectively. The intrauterine growth in the second half of pregnancy, evaluated using the transrectal and transabdominal approach, also showed strong positive correlations, similar to that reported for the mare. The trends of the combined thickness of the utero-placental unit and the echogenicity of the amniotic and allantoic fluids are examples. The diameters (mm) of fetal chest, eye orbit, and aorta increased throughout pregnancy and were 40.6 ± 2.9, 8.7 ± 1.5, and 3.5 ± 0.7, respectively, at week 13, and 190.9 ± 12.0, 21.4 ± 1.5, and 30.6 ± 1.8 at the last evaluation before parturition. In contrast, heart rate decreased as pregnancy progressed. Regression analyses between these parameters and day of gestation were statistically significant (P < 0.001). All fetuses consistently showed some intrauterine activity. Maternal plasma progestagens and estrone sulfate concentrations followed a pattern similar to that seen in mares, although the prepartal progestagen peak was lower in jennies. This study provides a range of ultrasonographic and endocrine values for normal pregnancy in jennies.     

Blood analysis in newborn donkeys: hematology,biochemistry, and blood gases analysis

The knowledge of reference ranges for hematologic, biochemical, and blood gas parameters in the different species and the influence of breed and age on them is a fundamental tool for the clinician. For this reason, the aim of this study was to evaluate the age-related changes of hematologic and biochemical parameters in Martina Franca donkey foals during the first 3 weeks of life and of blood gases during the first 24 hours of age. Fifteen healthy donkey foals were enrolled; blood samples were collected from each foal at 10 minutes after birth, 1 hour after the first and second suckles, 12 and 24 hours after birth, daily from Day 2 to 7, and at Days 10, 14, and 21 of life. Erythrocytes, leukocytes, and platelets counts were assessed; also metabolic (alanine aminotransferase, aspartate aminotransferase, gamma glutamyl transferase, creatinphospokinase, lactate dehydrogenase, alkaline phosphatase, glucose, blood urea nitrogen, creatinine, total proteins, albumins, cholesterol, and total bilirubin) and electrolytic parameters (Ca, P, Mg, Na, K, and Cl) were evaluated. Finally, blood gases and metabolic parameters (pH, pCO₂, pO₂, sO₂, TCO₂, HCO₃, lactate, and base excess) on venous blood were assessed with a portable analyzer. A statistical analysis to evaluate the influence of age and sex was performed. Several differences were found between sampling times, demonstrating that age influences these parameters. Moreover differences were found compared with data reported in literature for donkey foals of another species, horse foals, and adult donkeys. Although a great interindividual variation for some parameters exists, this study demonstrated that interval references should be addressed not only to different species, but also to specific breeds and to the neonatal period.     

Thyroid hormone concentrations in foals affected by perinatal asphyxia syndrome

The hypothalamus-pituitary-thyroid axis has specific functions, mostly related to metabolic activities, cell differentiation, and development. To the authors’ knowledge, there are no studies about thyroid hormone (TH) concentrations in foals affected by perinatal asphyxia syndrome (PAS). Hence, the aims of the study are (1) to evaluate plasma TH concentrations (T₃ and T₄) in healthy foals during the first 7 days of life; (2) to evaluate plasma TH concentration (T₃ and T₄) in critically ill foals affected by PAS during the first 7 days of hospitalization; and (3) to compare TH concentrations between surviving and nonsurviving critically ill foals. Forty-five Standardbred foals were enrolled in this prospective observational study: 21 healthy foals (group 1) and 24 foals affected by PAS (group 2). Jugular blood samples were collected within 10 minutes from birth/admission and every 24 hours for 7 days (t0–t7). TH concentrations were analyzed by RIA. In both groups, T₃ concentration was significantly lower at t4, t5, t6, and t7 compared with t1 (P < 0.05), and T₄ concentration was significantly higher at birth than at all other time points (P < 0.01). No differences were found in TH concentrations at admission between surviving (n = 20) and nonsurviving (n = 4) foals. Statistical comparison between healthy and PAS foals divided into age groups showed significantly lower TH concentrations at t0 in PAS foals <12 hours old at admission (P < 0.01). In conclusion, PAS may cause lower T₃ and T₄ concentrations in affected foals than in age-matched healthy foals, as reported for other systemic illnesses, such as sepsis and prematurity. TH concentrations showed no prognostic value, which maybe due to the small number of nonsurviving foals in this study. Further studies are needed to find out if thyroid replacement therapy could be useful in the treatment of critically ill foals affected by PAS.     

PGF(2α), LH, testosterone, oestrone sulphate, and cortisol plasma concentrations around sexual stimulation in jackass

Many hormones are involved in the regulation of male reproductive functions, controlling sexual behavior, and influencing sexual arousal, the onset of erection and ejaculation, and the post-ejaculatory detumescence. The aims of this study were to analyze the plasma concentrations of 15-ketodihydro-PGF_2α (PGFM), LH, testosterone (T), oestrone sulphate (OS), and cortisol (C) in relation to sexual stimulation and to evaluate the possible correlations among circulating hormones and between hormones and semen characteristics in the donkey stallion. Thirteen sexually experienced Martina Franca jackass of proven fertility were enrolled and semen was collected through an artificial vagina. Plasma samples were collected at 12, 9, 6 and 3 min before oestrous jenny exposure, at the first erection in the mating arena in the presence of an oestrous jenny, during ejaculation, at dismounting, 3, 6, 9 and 12 min after ejaculation in box, and then every 10 min during the following 50 min. PGFM showed an increasing trend with significant differences between the pre-ejaculatory and post-ejaculatory period, suggesting a role of this hormone in the control of ejaculation. LH showed a significantly higher concentration at ejaculation compared to last samples, while T showed significantly higher levels at erection, ejaculation and dismounting, probably for its influence on these processes and on sexual behavior. Finally, OS did not show any difference in the period of observation, while C presented a significant increase only 22 minutes after erection. The only hormonal correlation found was a positive one between LH and T at erection and dismounting, while T and OS were positively correlated with total and progressive motility, respectively.     

Characterization of the genetic diversity of <Emphasis Type="Italic">Mycobacterium tuberculosis</Emphasis> in São Paulo city,Brazil

Background

Tuberculosis is a major health problem in São Paulo, Brazil, which is the most populous and one of the most cosmopolitan cities in South America. To characterize the genetic diversity of Mycobacterium tuberculosis in the population of this city, the genotyping techniques of spoligotyping and MIRU were applied to 93 isolates collected in two consecutive years from 93 different tuberculosis patients residing in São Paulo city and attending the Clemente Ferreira Institute (the reference clinic for the treatment of tuberculosis).

Findings

Spoligotyping generated 53 different spoligotype patterns. Fifty-one isolates (54.8%) were grouped into 13 spoligotyping clusters. Seventy- two strains (77.4%) showed spoligotypes described in the international databases (SpolDB4, SITVIT), and 21 (22.6%) showed unidentified patterns. The most frequent spoligotype families were Latin American Mediterranean (LAM) (26 isolates), followed by the T family (24 isolates) and Haarlem (H) (11 isolates), which together accounted for 65.4% of all the isolates. These three families represent the major genotypes found in Africa, Central America, South America and Europe. Six Spoligo-International-types (designated SITs by the database) comprised 51.8% (37/72) of all the identified spoligotypes (SIT53, SIT50, SIT42, SIT60, SIT17 and SIT1). Other SITs found in this study indicated the great genetic diversity of M. tuberculosis, reflecting the remarkable ethnic diversity of São Paulo city inhabitants. The MIRU technique was more discriminatory and did not identify any genetic clusters with 100% similarity among the 93 isolates. The allelic analysis showed that MIRU loci 26, 40, 23 and 10 were the most discriminatory. When MIRU and spoligotyping techniques were combined, all isolates grouped in the 13 spoligotyping clusters were separated.

Conclusions

Our data indicated the genomic stability of over 50% of spoligotypes identified in São Paulo and the great genetic diversity of M. tuberculosis isolates in the remaining SITs, reflecting the large ethnic mix of the São Paulo city inhabitants. The results also indicated that in this city, M. tuberculosis isolates acquired drug resistance independently of genotype and that resistance was more dependent on the selective pressure of treatment failure and the environmental circumstances of patients.

     

Clinical use of dopamine antagonist sulpiride to advance first ovulation in transitional mares

Artificial photoperiod treatment is currently the best method to hasten the first ovulation of the breeding season in winter anoestrus mares. However, this is not easy to apply in large herds of mares and, to be effective, has to be planned in the northern hemisphere in December at the latest. Pharmacological treatments have been proposed as alternatives: GnRH agonists, progesterone or its synthetic agonist Altrenogest, and dopamino-antagonists, as pherphenazine, domperidone or sulpiride. Dopamino-antagonists protocols, beginning at a given day of the year, gave controversial results in terms of hastening ovulation. The aim of this study was to evaluate the efficacy of an up-to-21-d long dopamine antagonist (sulpiride) treatment on mares at the beginning of the spring transition for its ability to hasten estrous cyclicity. In Study 1, 49 seasonally-acyclic standardbred mares, maintained in paddocks under natural photoperiod, were treated with 1 mg/kg/d sulpiride at the evidence of the first follicle with of 25 mm in diameter until ovulation for a maximum of 21 d (Group S₁; n = 34) or left untreated (Group C₁; n = 15). Group S₁ and C₁ mares showed a follicle of 35 mm in diameter after 8 and 22 d (median; P < 0.05) and ovulated after 18 and 43 d, respectively (median; P < 0.05). Twenty-two/26 and 6/15 mares of the Group S₁ and C₁ ovulated within 30 d from the beginning of the treatment, respectively (P < 0.05). All the mares of the study cycled until Autumn, unless they became pregnant.In Study 2, pregnancy rates after the first ovulation of the year of 22 acyclic standardbred mares maintained in paddocks under natural photo-period, treated following the same protocol as Study 1 (S₂), and 47 untreated mares (C₂) were compared. In Groups S₂ and C_2, 63.6% and 61.7% of the mares became pregnant after the first cycle (P > 0.05) and 50.0% and 61.1% of the remaining became pregnant in the following cycles (P > 0.05), respectively.Beginning with sulpiride treatment when follicles were 25 mm in diameter resulted in a significant advancement of cyclicity in non-photo-stimulated mares. Pregnancy rates after artificial insemination of treated mares were similar to those of untreated animals.     

Morphological features of the placenta at term in the Martina Franca donkey

This study was designed to establish the morphological features of the placenta of the Martina Franca jenny. Ten placentas were harvested at the time of foal delivery and examined both for gross and histological characteristics. The following factors were determined: the total weight and volume of the placenta and its components, the surface area of the allantochorion, umbilical cord length and site of insertion, and the diameter of the umbilical cord vessels and vascular pattern type. The weight of the placenta was similar to previously reported for ponies, and represented 12% of foal birth weight. Umbilical cord length was similar to that in the horse but longer than in the pony, while cord weight was intermediate between the two. In a histological examination, numerous strong villi were observed at sites corresponding to the non-pregnant and pregnant horn and uterine body. No villi were detected in the area overlying the cervical star. Despite obvious similarities between the donkey and horse placenta, specific morphological features do exist, and are possibly related to the differences in length of gestation.