Interfacial concentrations of chloride and bromide and selectivity for ion exchange in vesicles prepared with dioctadecyldimethylammonium halides, lipids, and their mixtures

The local concentrations of chloride, Cl b, and bromide, Br b, in the interface of vesicles prepared with dioctadecyldimethylammonium chloride, DODAC, or bromide, DODAB, dipalmitoylphosphatidylcholine, DPPC, dimyristoylphosphatidylcholine, DMPC, and mixtures of DMPC, DPPC, and DODAC were determined by chemical trapping by analyzing product yields from spontaneous dediazoniation of vesicle-bound 2,6-dimethyl-4-hexadecylbenzenediazonium ion. The values of Cl b and Br b in DODAC and DODAB vesicles increase with vesicle size, in agreement with previous data showing that counterion dissociation decreases with vesicle size. Addition of tetramethylammonium chloride displaces bromide from the DODAB vesicular interface. The value for the selectivity constant for Br/Cl exchange at the DODAB vesicular interface obtained by chemical trapping was ∼2.0, well within values obtained for comparable amphiphiles. In vesicles of DPPC the values of Cl b were very sensitive to the nature of the cation and decreased in the order Ca 2+ > Mg 2+ > Li + > Na + > K + = Cs + = Rb + ≥ +. The effect of the cation becomes more important as temperature increases above the phase transition temperature, T m, of the lipid. The values of Cl b increased sigmoidally with the mol % of DODAC in vesicles prepared with DODAC/lipid mixtures. In sonicated vesicles prepared with DODAC and DMPC (or DPPC), the values of Cl b reach local concentrations measured for the pure amphiphile at 80 mol % DODAC. These results represent the first extensive study of local concentration of ions determined directly by chemical trapping in vesicles prepared with lipids, synthetic ampliiphiles, and their mixtures.

Structure of the pelvic and penile urethra - Relationship with the ducts of the sex accessory glands of the Mongolian gerbil (Meriones unguiculatus)

The urethra is the main place of entry for sexually transmitted pathogens. However, there is little literature on the morphology of the urogenital system, principally the urethra and ducts of the sex accessory glands. The Mongolian gerbil is an insectivorous, herbivorous and monogamous rodent with nocturnal habits; it has been used successfully as a laboratory animal since the 1960s. Therefore, the objective of the present paper was to describe the structure and ultrastructure of the urethra and its relations to the ducts of the accessory sex glands of the Mongolian gerbil (Meriones unguiculatus), contributing to the understanding of the reproductive biology of the rodent and aiming to provide data for future experimental studies. Conventional techniques of light and scanning electron microscopy were utilized. The urethra and ducts of the accessory sex glands are similar to those of the albino rat and the mouse. However, there is variation in drainage type among accessory sex glands for the inner urethra. The ducts of the seminal vesicle, the ductus deferens, drain their contents independently into the ampullary duct that opens in the urethra. The ducts of the prostate, coagulating and bulbourethral glands drain their contents independently into the urethra.

Desenvolvimento inicial e distribuição temporal de larvas e juvenis de Bryconamericus stramineus Eigenmann, 1908 (Osteichthyes, Characidae) na planície alagável do alto rio Paraná, Brasil

The aim of this paper was to analyze initial development and temporal distribution of larvae and juveniles of Bryconamericus stramineus and their relationship with some environmental variables of Ressaco do Leopoldo in the Upper Paraná River floodplain. The samples were taken from February 1991 to February 1992. The larvae and the juveniles were identified and described according to the with its morphologic and meristic characteristics. The larvae presented scarce pigmentation, subterminal mouth, medium anal opening and myomeres varying 35-40 (16-21 pre and IT-22 postanal). The greater densities of larvae and juveniles were verified in December 1991 and November 1991, respectively, mainly during the night. The larvae density was only correlated with precipitation.

Bryconamericus turiuba, a new species from the upper Rio Paraná system (Ostariophysi: Characiformes)

Bryconamericus turiuba, new species, is described from the upper Rio Paraná system in Brazil. It differs from all congeners in the following combination of characters: 15-20 branched anal-fin rays; four or five scales in a transverse series from the dorsal-fin origin to the lateral line; shallow body (depth 23.6-30.6% of SL); 37-43 perforated lateral line scales; first, third, and fifth or, more rarely, first and fourth outer premaxillary tricuspid teeth projecting anteriorly; two to five tricuspid to pentacuspid maxillary teeth; dentary with three or four tricuspid or pentacuspid large teeth, followed by five to seven smaller conical to tricuspid teeth; a conspicuous dark, vertically elongate humeral spot, extending to below the lateral line; a dark dorsal stripe extending from the supraoccipital spine to the caudal peduncle with a gap at the adipose-fin base; caudal-fin lobes without conspicuous markings, uniformly pigmented along rays; mature males lacking hooks on the pelvic- and anal-fin rays. The new species is compared with the Bryconamericus species described from the basins of the Rio Paraná, Rio Paraguay, Rio São Francisco, and coastal Brazilian rivers. © 2005 by the American Society of Ichthyologists and Herpetologists.

Fertilização e desenvolvimento embrionário: Morfometria e análise estereomicroscópica dos ovos dos híbridos de surubins (pintado, Pseudoplatystoma corruscans x cachara, Pseudoplatystoma fasciatum)

This study aimed to analyze the morphometry of hybrids' eggs under stereomicroscopy. The induced reproduction was carried out in specimens of pintado's females (P. corruscans) and cachara's males (P. fasciatum) from Cepta - Ibama, Pirassununga, state of São Paulo, Brazil. Samples were collected in extrusion moment, during the fertilization, in times of 10 and 30 seconds, 1, 2, 5, 8, 10, 15, 20, 30 and 45 minutes, 1 hour, at every 15 minutes until completing 2 hours, and afterwards, at every hour until the larval hatching. Oocytes and eggs' external morphology were observed, photographed and 30 samples from each moment were measured in stereomicroscopy. Oocytes, in the time of extrusion, and the eggs presented a spherical form and yellowish color. Characteristics such as wide perivitelinic space, somites, presence of optical vesicle and the main phases of embryonic development as egg-cell (blastodisc), 2, 4, 8, 16, 32 and 64 blastomeres, morula, blastula, gastrula, epiboly movements, formation and larvae hatching were verified.

The reproductive biology and cycle of Rioraja agassizi (Müller and Henle, 1841) (Chondrichthyes: Rajidae) in southeastern Brazil, SW Atlantic Ocean

Specimens of Rioraja agassizi were collected at Guarujá pier, São Paulo State, Brazil, from March 2005 to March 2006. A total of 275 males were captured. Based on the analysis of the clasper length, gonad weight, clasper gland length, alar thorns and lobule diameter, the size-at-maturity was estimated to be 32 cm. There were 1049 females in the record. Size-at-maturity was calculated in 40 cm. Sexual resting females were observed. Gonadosomatic and hepatosomatic indexes varied significantly in both sexes throughout the year. However, ovulation, egg-laying and presence of sperm in the seminal vesicle were observed all year round. For this reason, an annual cycle with at least one peak in the sexual activity is proposed for this species.

Prevalência de papiloma vírus (HPV) perianal assintomático em pacientes portadores de HPV genital tratados no Hospital das Clínicas da Faculdade de Medicina de Botucatu

Introduction: Human Papillomavirus (HPV) infection is one of the Sexually Transmitted Diseases (STD) of greatest incidence and prevalence worldwide, and it is presently considered to be a pre-neoplastic lesion. Human Papillomavirus infection has totaled 23.4% of the sexually transmitted diseases reported to the Department of Health, and it is currently the most common in our country. Many patients are asymptomatic carriers. Methods: Twenty patients who had been previously treated for HPV due to genital lesions were referred for the conduction of anuscopy and colposcopy of the perianal region. Results: Males (80%) prevailed over females (20%). Of the total number of patients, only 2 showed lesions as examined by anuscopy (10%). However, 3 other patients showed lesions by means of colposcopy, thus increasing to 5 (25%) the total number of asymptomatic patients who presented perianal lesions. Of the total number of patients with lesions, 4 were males and 1 was a female. Conclusion: The presence of perianal lesions was observed in 10% of the patients with genital lesions by means of simple anal inspection. This figure increased to 25% when anal colposcopy was associated, thus showing the importance of conducting such examination on all patients with increased risk factors for HPV infection in the anal region.

Localization of myosin-Va in subpopulations of cells in rat endocrine organs

Myosin-Va is a Ca 2+/calmodulin-regulated unconventional myosin involved in the transport of vesicles, membranous organelles, and macromolecular complexes composed of proteins and mRNA. The cellular localization of myosin-Va has been described in great detail in several vertebrate cell types, including neurons, melanocytes, lymphocytes, auditory tissues, and a number of cultured cells. Here, we provide an immunohistochemical view of the tissue distribution of myosin-Va in the major endocrine organs. Myosin-Va is highly expressed in the pineal and pituitary glands and in specific cell populations of other endocrine glands, especially the parafollicular cells of the thyroid, the principal cells of the parathyroid, the islets of Langerhans of the pancreas, the chromaffin cells of the adrenal medulla, and a subpopulation of interstitial testicular cells. Weak to moderate staining has been detected in steroidogenic cells of the adrenal cortex, ovary, and Leydig cells. Myosin-Va has also been localized to non-endocrine cells, such as the germ cells of the seminiferous epithelium and maturing oocytes and in the intercalated ducts of the exocrine pancreas. These data provide the first systematic description of myosin-Va localization in the major endocrine organs of rat. © 2008 Springer-Verlag.

The role of membranous urethral afferent autonomic innervation in the continence mechanism after nerve sparing radical prostatectomy: a clinical and prospective study

Purpose: We evaluated the somatic and autonomic innervation of the pelvic floor and rhabdosphincter before and after nerve sparing radical retropubic prostatectomy using neurophysiological tests and correlated findings with clinical parameters and urinary continence. Materials and Methods: From February 2003 to October 2005, 46 patients with prostate cancer were enrolled in a controlled, prospective study. Patients were evaluated before and 6 months after nerve sparing radical retropubic prostatectomy using the UCLA-PCI urinary function domain and neurophysiological tests, including somatosensory evoked potential, and the pudendo-urethral, pudendo-anal and urethro-anal reflexes. Clinical parameters and urinary continence were correlated with afferent and efferent innervation of the membranous urethra and pelvic floor. We used strict criteria to define urinary continence as complete dryness with no leakage at all, not requiring any pads or diapers and with a UCLA-PCI score of 500. Patients with a sporadic drop of leakage, requiring up to 1 pad daily, were defined as having occasional urinary leakage. Results: Two patients were excluded from study due to urethral stricture postoperatively. We evaluated 44 patients within 6 months after surgery. The pudendo-anal and pudendo-urethral reflexes were unchanged postoperatively (p = 0.93 and 0.09, respectively), demonstrating that afferent and efferent pudendal innervation to this pelvic region was not affected by the surgery. Autonomic afferent denervation of the membranous urethral mucosa was found in 34 patients (77.3%), as demonstrated by a postoperative increase in the urethro-anal reflex sensory threshold and urethro-anal reflex latency (p <0.001 and 0.0007, respectively). Six of the 44 patients used pads. One patient with more severe leakage required 3 pads daily and 23 showed urinary leakage, including 5 who needed 1 pad per day and 18 who did not wear pads. Afferent autonomic denervation at the membranous urethral mucosa was found in 91.7% of patients with urinary leakage. Of 10 patients with preserved urethro-anal reflex latency 80% were continent. Conclusions: Sensory and motor pudendal innervation to this specific pelvic region did not change after nerve sparing radical retropubic prostatectomy. Significant autonomic afferent denervation of the membranous urethral mucosa was present in most patients postoperatively. Impaired membranous urethral sensitivity seemed to be associated with urinary incontinence, particularly in patients with occasional urinary leakage. Damage to the afferent autonomic innervation may have a role in the continence mechanism after nerve sparing radical retropubic prostatectomy.

Perinatal exposure to parasiticide ivermectin: Long-term effects on reproductive parameters of adult male rats

Ivermectin is one of the most widely used antiparasitic drugs globally. The aim of this study was to evaluate the chronic effects of perinatal exposure to ivermectin on male reproductive parameters in rats. Pregnant rats were treated daily by oral gavage with 0.4 or 1.6 mg kg -1 of ivermectin or vehicle, from gestational day 6 until post-natal day 10. In the adulthood stage, there were significant reductions in the relative testicular weight of rats exposed to the low dose and in relative prostate weight of male rats exposed to the high dose of ivermectin. Furthermore, the animals exposed to the low dose also presented an increased seminal vesicle weight compared to controls. However, neither of the ivermectin doses interfered in daily sperm production, sperm number in testis, or sexual behavior of exposed males. In conclusion, perinatal exposure to ivermectin neither altered the male reproductive system development markedly, nor produced any adverse effects on the parameters evaluated. © 2011 Taylor & Francis.

Description and genetic analysis of three sets of monozygotic twins resulting from transfers of single embryos to recipient mares

Case Description-3 sets of monozygotic twins resulting from transfers of single embryos to recipient mares were examined. Clinical Findings-In all 3 recipient mares with twin pregnancies, only 1 embryonic vesicle was detected before day 25 of gestation. In 1 recipient mare, 2 apparent adjacent vesicles, each containing an embryo with a heartbeat, were visualized on ultrasonographic examination on day 37 of gestation. The other 2 recipient mares underwent ultrasonographic examination on day 30 of gestation, at which time only 1 vesicle and embryo was identified. In these latter 2 recipient mares, however, a thorough ultrasonographic examination for a second conceptus on day 30 had not been performed, as only 1 embryo had been transferred and visualized on early ultrasonographic examination. Treatment and Outcome-All twin pregnancies resulted in death of both fetuses. Genetic analysis confirmed that each set of monozygotic twins originated from the transferred embryo. Clinical Relevance-Monozygotic twin pregnancy may occur after embryo transfer; thus recipient mares should be examined thoroughly for multiple conceptuses, especially between 25 and 30 days of gestation. At this time, the allantoides of monozygotic twins should be visible ultrasonographically and effective management may still be possible.

Spermatogenesis of riffle bugs, Rhagovelia whitei and Rhagovelia sp (Veliidae), and backswimmers Martarega sp (Notonectidae).

We examined the course of spermatogenesis and the meiotic chromosome complements in aquatic species of true bugs, Heteroptera. The chromosome complement of the Veliidae species was 2n = 39 (38A + X0) and 23 (22A + X0) in Rhagovelia whitei and Rhagovelia sp, respectively, and in the species of the Notonectidae (Martarega sp) it was 26 (22A + 2m + XY); all collected from the region of São José do Rio Preto, SP, Brazil. An impressive characteristic of the first analysis was the size of the cells belonging to Martarega sp, which were six times larger than the same cells in Pentatomidae and twice as large as the cells in aquatic Heteroptera (Gerridae). Regarding spermatogenesis, all the species analyzed showed the same pattern: holocentric chromosomes and elongated spermatids with the chromatin distributed evenly along the head. The family Veliidae showed several bodies impregnated with silver nitrate at prophase, while the family Notonectidae displayed only one. The cells of Notonectidae also showed an evident and round body until the end of prophase I and in the family Veliidae the silver-impregnated bodies were disorganized, where the only region visualized was possibly that of the NOR. In metaphase, silver-stained regions were found at the periphery of all chromosomes in Veliidae and at the periphery of some chromosomes in Notonectidae. The spermatids of Veliidae showed a less silver-impregnated vesicle, while Notonectidae showed silver staining only in part of the nuclear membrane. Therefore, families of Heteroptera have some differences and features that can help identify and classify these species.

Physiology and endocrinology symposium: Influence of cattle genotype (Bos indicus vs. Bos taurus) on oocyte and preimplantation embryo resistance to increased temperature

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Vitrification of immature feline oocytes with a commercial kit for bovine embryo vitrification

The aim of this study was to evaluate the suitability of a commercial kit for bovine embryo vitrification for cryopreserving cat oocytes and to evaluate comparatively the effects of its use with slow freezing procedure on cryotolerance in terms of morphology and oocyte resumption of meiosis. Germinal vesicle stage oocytes isolated from cat ovaries were either vitrified (n=72) using a vitrification kit for bovine embryo or slow frozen (n=69) by exposing oocyte to ethylene glycol solution before being transferred to a programmable embryo freezer. After thawing and warming, oocytes were cultured for 48h and then were examined for meiosis resumption using bisbenzimide fluorescent staining (Hoechst 33342). Fresh immature oocytes (n=92) were used as the control group. The proportion of oocytes recovered in a morphologically normal state after thawing/warming was significantly higher in frozen oocytes (94.5%) than in the vitrified ones (75%, p<0.01). Morphological integrity after culture was similar in vitrified (73.6%) and slow frozen oocytes (76.8%); however, only 37.5% of the morphologically normal oocytes resumed meiosis after vitrification compared to 60.9% of those submitted to slow freezing procedure (p<0.01). Fresh oocytes showed higher morphological integrity (91.3%) and meiosis resumption rates (82.6%, p<0.002) than cryopreserved oocytes, irrespective of the procedure used. These results suggest that immature cat oocytes vitrified with a kit for bovine embryos retain their capacity to resume meiosis after warming and culture, albeit at lower rates than slow frozen oocytes. Vitrification and slow freezing methods show similar proportions of oocytes with normal morphology after culture, which demonstrate that thawed and warmed oocytes that resist to cryodamage have the same chances to maintain their integrity after 48h of culture. © 2012 Blackwell Verlag GmbH.

Ovarian morphology and oogenesis of Leptodesmus dentellus (Diplopoda, Polydesmida, Chelodesmidae)

This study presents the ovarian morphology and the dynamics of the vitellogenesis process in the oocytes of the diplopod Leptodesmus dentellus. The oocytes are arranged in clusters called ovisacs which are distributed in pairs along the midline of the body forming the ovary. Regions similar to the germarium appear paired in the anterior region of the ovary; however, the development of the oocytes of this species does not follow a regionalisation in the reproductive organ. Cells in three developmental stages are found throughout the length of the ovary. Calcium, proteins, lipids and neutral polysaccharides were detected in the oocytes of L. dentellus. The polysaccharides and the proteins found in the oocytes have a double origin: endogenous, with the participation of the germinative vesicle, and exogenous, from follicular epithelium. The origin of the lipids is exogenous, i.e., they are incorporated into the oocytes, probably derived from the perivisceral fat body, which are highly developed in this region. The deposition of calcium is pre-vitellogenic and probably functions as a reserve during the juvenile stages. © 2012 Koninklijke Brill NV, Leiden.