Rodrigo Alves de Oliveira

Sperm Protamine-Status Correlates to the Fertility of Breeding Bulls

ABSTRACT During fertilization, spermatozoa make essential contributions to embryo development by providing oocyte activating factors, centrosomal components, and paternal chromosomes. Protamines are essential for proper packaging of sperm DNA; however, in contrast to the studies of oocyte-related female infertility, the influence of sperm chromatin structure on male infertility has not been evaluated extensively. The objective of this study was to determine the sperm chromatin content of bull spermatozoa by evaluating DNA fragmentation, chromatin maturity/protamination, PRM1 protein status, and nuclear shape in spermatozoa from bulls with different fertility. Relationships between protamine 1 (PRM1) and the chromatin integrity were ascertained in spermatozoa from Holstein bulls with varied (high vs. low) but acceptable fertility. Sperm DNA fragmentation and chromatin maturity (protamination) were tested using Halomax assay and toluidine blue staining, respectively. The PRM1 content was assayed using Western blotting and in-gel densitometry, flow cytometry, and immunocytochemistry. Fragmentation of DNA was increased and chromatin maturity significantly reduced in spermatozoa from low-fertility bulls compared to those from high-fertility bulls. Field fertility scores of the bulls were negatively correlated with the percentage of spermatozoa displaying reduced protamination and fragmented DNA using toluidine blue and Halomax, respectively. Bull fertility was also positively correlated with PRM1 content by Western blotting and flow cytometry. However, detection of PRM1 content by Western blotting alone was not predictive of bull fertility. In immunocytochemistry, abnormal spermatozoa showed either a lack of PRM1 or scattered localization in the apical/acrosomal region of the nuclei. The nuclear shape was distorted in spermatozoa from low-fertility bulls. In conclusion, we showed that inadequate amount and localization of PRM1 were associated with defects in sperm chromatin structure, coinciding with reduced fertility in bulls. These findings are highly significant because they reveal molecular and morphological phenotypes of mammalian spermatozoa that influence fertility.

Visceral leishmaniasis after renal transplantation : report of 4 cases in northeastern Brazil

Abstract: Visceral leishmaniasis (VL) is a well recognized opportunistic infection in immunosuppressed patients, which may cause febrile illness. We describe 4 renal transplant patients with VL in an endemic area in Brazil and their response to therapy. In 3 cases the diagnosis was confirmed by bone marrow aspirate that revealed the presence of Leishmania. In 1 case the bone marrow aspirate was inconclusive and the diagnosis was made through spleen biopsy that showed the presence of the parasite. VL needs to be considered as a cause of febrile illness in transplanted patients living in endemic areas.

Renal tubular dysfunction in patients with American cutaneous leishmaniasis.

Renal dysfunction seen in patients with American cutaneous leishmaniasis (ACL) has been attributed to the use of antimonials for treatment. To determine whether ACL itself causes tubular dysfunction, we measured renal function in 37 patients with ACL prior to their treatment and compared results to that in 10 healthy volunteers of similar mean age. None of the patients presented with glomerular dysfunction; however, 27 had a urinary concentrating defect. There was no statistical difference between groups in the pre- and post-desmopressin test of urine osmolality, but the post-test urine osmolality of the controls was significantly higher. Urinary AQP2 levels, determined by western blot of isolated exosomes, were found to be significantly lower in patients than in controls, whereas that of the cotransporter (NKCC2) was significantly higher. A urinary acidification defect (post-test pH greater than 5.50 following calcium chloride) was found in 15 patients. Pretest plasma bicarbonate was below normal in 12 patients as was the pretest plasma pH in 14. Expression of the Na/H exchanger (NHE3), H(+)-ATPase, and pendrin were all significantly higher in patients with ACL than in controls. A combined urinary concentration and acidification defect was found in 12 patients. Thus, the urinary concentrating defect of ACL may be caused by decreased AQP2, with increased NKCC2 compensatory. Pendrin upregulation may be related to the urinary acidification defect with increased NHE3 and H(+)-ATPase also compensatory. Hence, ACL can cause asymptomatic renal tubular dysfunction.

Acute renal failure following bee stings: case reports

Rhabdomyolysis is a syndrome characterized by muscle injury, most frequently due to muscle crushing and trauma. However, it may also be induced by non-traumatic causes, for example by means of stinging by Africanized bees. We describe two cases of rhabdomyolysis that presented dialytic acute renal failure after several bee stings.

Renal function evaluation in patients with American Cutaneous Leishmaniasis after specific treatment with pentavalent antimonial

BackgroundRenal evaluation studies are rare in American Cutaneous Leishmaniasis (ACL). The aim of this study is to investigate whether specific treatment reverts ACL-associated renal dysfunction.MethodsA prospective study was conducted with 37 patients with ACL. Urinary concentrating and acidification ability was assessed before and after treatment with pentavalent antimonial.ResultsThe patients mean age was 35.6 ± 12 years and 19 were male. Before treatment, urinary concentrating defect (U/Posm <2.8) was identified in 27 patients (77%) and urinary acidification defect in 17 patients (46%). No significant glomerular dysfunction was observed before and after specific ACL treatment. There was no reversion of urinary concentrating defects, being observed in 77% of the patients before and in 88% after treatment (p = 0.344). Urinary acidification defect was corrected in 9 patients after treatment, reducing its prevalence from 40% before to only 16% after treament, (p = 0.012). Microalbuminuria higher than 30 mg/g was found in 35% of patients before treatment and in only 8% after treatment. Regarding fractional excretion of sodium, potassium, calcium, phosphorus and magnesium, there was no significant difference between pre and post-treatment period.ConclusionAs previously described, urinary concentrating and acidification defects were found in an important number of patients with ACL. Present results demonstrate that only some patients recover urinary acidification capacity, while no one returned to normal urinary concentration capacity.

Fatal disseminated strongyloidiasis after kidney transplantation

Disseminated strongyloidiasis (DS) is a rare and severe parasitic disease that is difficult to recognize and affects immunocompromised individuals. We report the case of a kidney transplant recipient who presented with DS despite prophylaxis with albendazole. We have discussed the need for better prophylactic strategies and for a higher degree of suspicion in order to diagnose DS.

Performance of Orius insidiosus on alternative foods

Orius insidiosus is economically important for biological control of pests of vegetable and ornamental crops. To improve pest control with this predator, its densities in the crops can be enhanced by the provision of alternative foods, especially when prey are scarce. We therefore compared the performance of O. insidiosus on three alternative foods (Ricinus sp. pollen, bee pollen and the mixed stages of the astigmatid prey Tyrophagus putrescentiae) that are cheaper than frozen eggs of Ephestia kuehniella. Juvenile development was significantly shorter on E. kuehniella and T. putrescentiae than on Ricinus sp. pollen and on bee pollen and lowest without food. Female bugs had a higher oviposition rate when fed either E. kuehniella or T. putrescentiae, but produced fewer eggs when feeding on bee pollen and Ricinus sp. pollen. This shows that T. putrescentiae can possibly be used as cheap alternative food to boost predator populations in periods of low prey densities.

109 ASSESSMENT OF THE FUNCTIONALITY OF PLASMA MEMBRANE OF CRYOPRESERVED SPERM FROM DIFFERENT BREEDS OF SHEEP

Sperm cryopreservation is very important to genetic material quality dissemination. However, the spermatozoa can be damaged during this process. The objective of this study was to investigate the functionality of spermatozoa plasmatic membrane of ovine after cryopreservation. The work was conducted at Laboratory of Reproduction Biotechnology, Veterinary Medicine course, Federal University of Piaui. Fifteen animals were used (5 Dorper, 5 Santa Ines, and 5 animals of undefined breed - SRD). The animals were between 2 and 3 years old, and only one ejaculate was used per animal. Semen was collected via artificial vagina and frozen in cryopreservation machine (TK 3000®, OV curve 2), with Tris-yolk base extend and glycerol. The semen was evaluated before and after cryopreservation. To investigate the functionality of spermatozoa plasmatic membrane hypo-osmotic test (HOST) was used. After thawing, 20 μL of semen was incubated in 2 mL of hypoosmotic solution (125 mOsmol L-1) and 20 μL of semen was incubated in a 4% formalide citrate solution at 37°C for 1 h. After incubations, both samples were placed on microscopic slides, and 200 cells were visually evaluated per slide. Data were analyzed by Tukey-Kramer test (parametric data) and Dunn test (no parametric data) by Grafpad Instat® software. The motility averages after thawing were 36 ± 15.2% (Dorper), 36 ± 19.5% (Santa Ines), and 21 ± 14.3% (SRD). If the plasma membrane over the principal piece is intact, the membrane will swell, causing the tail to coil, whereas spermatozoa with damaged principal piece membranes will not swell. The HOST averages were 24.2 ± 11.8% (Dorper), 21% ± 6.4 (Santa Ines), and 10.4 ± 1.6% (SRD). Motility and HOST showed positive correlation (P < 0.001). The SRD animals showed higher correlation numbers than other groups (P < 0.009). The HOST is a cheap, easy, and efficient technique to estimate the functionality of sperm plasmatic membrane. We suggest that other studies could be done with more animals and more samples.