Aspectos da biologia reprodutiva de Boa constrictor constrictor: um estudo histológico dos testículos nos períodos reprodutivos de quiescência e máxima atividade / Boa constrictor constrictor reproductive biology aspects: the testes histological study in the reproductive periods of quiescence and maximal activity

As serpentes pertencem ao segundo maior grupo dentro dos répteis, podendo apresentar sazonalidade quanto à espermatogênese, com produção descontínua ou contínua. O presente trabalho tem como objetivo caracterizar aspectos da biologia reprodutiva de Boa constrictor constrictor com base nos achados histológicos dos testículos nos períodos de máxima atividade (período de gametogênese) e quiescência. Os testículos de dois espécimes de Boa c. constrictor (7767 e 11752) foram cortados a uma espessura de 3µm em micrótomo, corados com azul de toluidina 1%, fotodocumentados e descritos. A presença de espermatozoides na luz do túbulo seminífero no indivíduo 7767 indica um período de máxima gametogênese, enquanto o lúmen dos túbulos seminíferos pouco evidentes, sem a presença de espermatozoides e de células gaméticas em divisão, caracteriza o indivíduo 11752 em período quiescente. Mediante os achados histológicos descritos no presente estudo, concluiu-se que Boa c. constrictor apresenta sazonalidade em relação à gametogênese, sendo esse padrão de sazonalidade associado ao período de cópulas relatado em literatura característico de serpentes com padrão pré-nupcial.(AU)

These snakes belong to the second largest group within the reptiles, being able to present seasonality regarding spermatogenesis, with discontinuous or continuous production. The present study aims to characterize Boa constrictor constrictor reproductive biology aspects from histological findings in testicles during periods of maximum activity (period of gametogenesis) and quiescence. The testicles of two specimens of Boa c. constrictor (7767 and 11752) were cut to a thickness of 3µm in microtome, stained with 1% toluidine blue, photodocumented and described. The spermatozoa presence in the seminiferous tubule lumen in individual 7767 indicates a period of maximum gametogenesis, whereas the seminiferous tubules lumen is not very evident without spermatozoa and the absence of dividing gametic cells characterizes individual 11752 in the quiescent period. Through the histological findings we concluded that Boa c. constrictor presents seasonality in relation to gametogenesis, and the pattern of reproductive seasonality observed along with the period of copulas reported in the literature resembles the pre-nuptial pattern.(AU)

Disintegration time of orally dissolving films: various methodologies and in-vitro/in-vivo correlation.

Orodispersible films (ODF) have gained a remarkable success in the market, especially in pediatric and geriatric populations. The time required for an ODF to disintegrate is an essential quality and safety feature, thus an appropriate methodology is crucial. The USP disintegration apparatus is not appropriate for ODFs, as the determination of the end point is challenging and may not predict in-vivo disintegration time. The aim of the present study was to design and evaluate new disintegration protocols as an attempt to select the best approach that would reflect the in-vivo disintegration time in comparison to formerly reported procedures. Novel methods were designed, namely; the frame, the cell, and the agar plate methods, and compared to the previously reported methods; clamp and modified USP disintegration methods. Different ODFs were formulated using various viscosity grades of hydroxypropylmethyl cellulose. The mechanical characteristics of the prepared films were studied using texture analyzer and film folding endurance test. The resultant disintegration time of the films measured by the aforementioned methods were compared and correlated with its in-vivo time. Interestingly, the results obtained through the use of the cell method for the low viscosity polymers did not vary significantly from that of their in-vivo results (p>0.05). Moreover, the disintegration time of all polymeric films determined by the cell method revealed independently on their viscosity the highest correlation with in-vivo disintegration time (R² = 0.999). Such findings indicated the suitability of the cell method in predicting in-vivo disintegration time of low viscosity polymeric films.

Elucidation of the molecular mechanism underlying the anti-inflammatory activity of an effective and safe bipyrazole-based compound.

INTRODUCTION: Since the synthesis of acetylsalicylic acid by Hoffmann in 1897, new classes of NSAIDs have been introduced; however, their side effects have limited their clinical applications. Consequently, our team has recently synthesized a novel bipyrazole compound that showed a satisfactory efficacy and safety profile. The aim of the current study was to elucidate the molecular mechanism of this bipyrazole compound. METHOD: The anti-inflammatory efficacy of the compound was assessed using formalin-induced paw edema test. Computer-assisted simulation docking experiments were carried out. Cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), neuronal nitric oxide synthase (nNOS), tumor necrosis factor-alpha (TNFα), interleukin-1 (IL1) and interleukin-10 (IL10) gene expression were quantified with real-time polymerase chain reaction (RT-PCR) using SYBR Green technology. The samples were taken from the plantar paw of mice after formalin local injection. RESULTS: The efficacy of the bipyrazole compound was similar to that of indomethacin, diclofenac, and celecoxib, as proven by the formalin-induced paw edema. Docking study indicated a superior binding score for the studied compound relative to celecoxib, indomethacin, and diclofenac. RT-PCR assessment revealed a significant decrease in iNOS, COX-2, and TNFα gene expression in the bipyrazole-treated group. Moreover, a reduction in IL1 and nNOS gene expression levels and an increase in IL10 level were detected despite being insignificant compared to the control group. CONCLUSION: These findings revealed the superiority of the newly synthesized bipyrazole compound not only on the binding site, but also by inhibiting most of the inflammatory mediators including TNF-α.

Biochemical markers as diagnostic/prognostic indicators for ischemic disease.

OBJECTIVE: The use of a biomarker was extremely useful in clinical emergencies such as stroke to aid in triage and early management of cases. The diagnostic accuracy of laboratory biomarkers is run to approve the identification of easy, cheap and fast tests associated with cerebral ischemia and intracranial hemorrhage. The present study was designed to screen serum enolase activity, activities of CK-BB, LDH and lipid profile in patients with ischemic or related diseases as good diagnostic/ prognostic indicator for ischemic diseases. METHODS: Sixty male subjects in the age range of (45 ±2years) were divided into four groups each with 15 participants: Group (I) normal . Group (II) patients recently diagnosed as ischemic disease; Group (III) hypertensive patients and Group (IV); diabetic patients enolase activity (p<0.001) and CK-BB (p<0.01) in ischemic and hypertensive patients compared with control and diabetic groups. LDH level was significantly elevated in ischemic, hypertensive and diabetic patients compared with controls (p<0.001). The cut -off value for serum enolase was 62.5 nmol/l showing 90% sensitivity and 93% specificity for differentiation of ischemic disease. Positive correlations were observed between serum enolase (r = 0.56), and CK-BB (r = 0.53). CONCLUSION: Serum enolase can be considered as a more sensitive and specific marker and used as a sensitive diagnostic or prognostic marker for ischemic related diseases.

Morphologic and morphometric analysis of testis of Pseudis limellum (Cope, 1862) (Anura, Hylidae) during the reproductive cycle in the Pantanal, Brazil

The spermatogenesis of Pseudis limellum, from the Southern Pantanal, Brazil, was studied from July 1995 to May 1996, through histological sections of the testis. The cells could be differentiated as: primary spermatogonia, large cells, generally with bilobed nucleus; secondary spermatogonia, smaller cells, with darker cytoplasm, chromatin of radial form; primary and secondary spermatocytes, differentiated according to the different stages of the nucleus during the successive cells divisions. Furthermore, we observed cells in process of morphologic differentiation: rounded spermatids much smaller, with nucleus containing chromatin in compacting process and cytoplasm reduction; elongated spermatids, generally parallel organized in well defined bundles, with the anterior region directed toward the periphery of the seminiferous tubule and the tail directed toward the lumen. Spermatozoa are free in the lumen of the seminiferous tubule. All the cells are organized as cysts, which are supported by a large amount of Sertoli cells. The spermatogenesis in P. limellum is very similar to that of other anurans, but peculiarities were observed regarding the organization of the germ cells, the great amount of free Sertoli cells in the lumen of testis collected in May, and the long cytoplasmatic extensions of the cells bearing pigments and involving the seminiferous tubule. The diameter of the seminiferous tubule (SD) exhibited an annual mean of 251.79 +/- 37.57 microm. Spermatozoa number by seminiferous tubule (SN) exhibited an annual mean of 306.66 +/- 39.83, also with higher and lower values at each month. Variations in SD and SN were not significantly correlated with climatic variables. In this species, reproduction occurs throughout the year in ponds and flooded areas, despite the seasonal climate of the Pantanal. Although males varied in their annual reproductive activity, they were considered potentially reproductive in all months throughout the year.(AU)

Morphologic and morphometric analysis of testis of Pseudis limellum (Cope, 1862) (Anura, Hylidae) during the reproductive cycle in the Pantanal, Brazil

The spermatogenesis of Pseudis limellum, from the Southern Pantanal, Brazil, was studied from July 1995 to May 1996, through histological sections of the testis. The cells could be differentiated as: primary spermatogonia, large cells, generally with bilobed nucleus; secondary spermatogonia, smaller cells, with darker cytoplasm, chromatin of radial form; primary and secondary spermatocytes, differentiated according to the different stages of the nucleus during the successive cells divisions. Furthermore, we observed cells in process of morphologic differentiation: rounded spermatids much smaller, with nucleus containing chromatin in compacting process and cytoplasm reduction; elongated spermatids, generally parallel organized in well defined bundles, with the anterior region directed toward the periphery of the seminiferous tubule and the tail directed toward the lumen. Spermatozoa are free in the lumen of the seminiferous tubule. All the cells are organized as cysts, which are supported by a large amount of Sertoli cells. The spermatogenesis in P. limellum is very similar to that of other anurans, but peculiarities were observed regarding the organization of the germ cells, the great amount of free Sertoli cells in the lumen of testis collected in May, and the long cytoplasmatic extensions of the cells bearing pigments and involving the seminiferous tubule. The diameter of the seminiferous tubule (SD) exhibited an annual mean of 251.79 +/- 37.57 microm. Spermatozoa number by seminiferous tubule (SN) exhibited an annual mean of 306.66 +/- 39.83, also with higher and lower values at each month. Variations in SD and SN were not significantly correlated with climatic variables. In this species, reproduction occurs throughout the year in ponds and flooded areas, despite the seasonal climate of the Pantanal. Although males varied in their annual reproductive activity, they were considered potentially reproductive in all months throughout the year.

Effect of Norplant implants on the pituitary-adrenal axis function and reserve capacity.

The present work investigated the effects of Norplant implants on the pituitary-adrenal function among 15 users of Norplant implants prior to and 6 months after insertion of the implants. Serum cortisol levels and their diurnal variations, ACTH and 24-h urinary 17-ketosteroids, ketogenic steroids, 17-hydroxy steroids, and creatinine, were measured. Also, a dynamic test (the 5-h Synacthen depot = ACTH stimulation test) was done before and 6 months after implants insertion. The 9 a.m. cortisol levels were blunted (within the normal ranges) while the 6 p.m. values were unaltered. The 24-h urinary ketogenic, hydroxy, and ketosteroids were also unchanged after Norplant implants use. The ACTH stimulation test showed a decreased adrenal response which was also within normal ranges. These data should raise the question related to suprarenal response to acute or prolonged stresses, such as surgical operations or shock in women using Norplant implants.

PIP: To ensure that Norplant contraceptive implants are not associated with a risk of pituitary-adrenal suppression, a series of laboratory tests were conducted in 15 women both before and 6 months after Norplant insertion. Comparisons of hormonal profiles before and after Norplant insertion revealed a significant drop in morning serum cortisol levels (404.33 +or- 84.07 nmol/l vs. 353.67 +or- 56.65 nmol/l, p 0.05), but no significant change in evening readings. The observed changes in morning cortisol values were still within the normal range. Serum ACTH values and 24-hour urinary 17-hydroxy steroids, 17-ketogenic steroids, and 17-ketosteroids were not different after insertion compared to baseline. Before Norplant insertion, injection of synthetic ACTH resulted in a 259.59 +or- 169.53% increase in the mean level of serum cortisol 5 hours later; 6 months after Norplant insertion, the percent rise above baseline was 165.85 +or- 91.64%. The significantly lower adrenal response among Norplant users (although still within normal limits) is presumably due to a local inhibition of the adrenal itself and not of the hypothalamic-pituitary axis. Although these findings suggest a minimal suppressive effect of prolonged microdose release of levonorgestrel from Norplant implants, the suprarenal response to acute or prolonged stresses (e.g., surgical operations or shock) in Norplant users requires investigation.

Chlamydial serology among patients with tubal factor infertility and ectopic pregnancy in Alexandria, Egypt.

BACKGROUND AND OBJECTIVES: Little is known about the role of Chlamydia trachomatis in the etiology of tubal factor infertility and ectopic pregnancy in Egypt. GOAL OF THIS STUDY: To assess the association between past chlamydial infection, tubal factor infertility, and ectopic pregnancy in an Egyptian population. STUDY DESIGN: This report consists of two concurrent case-control studies. First, 51 patients with tubal factor infertility were compared with 48 healthy subjects who did not have tubal factor infertility and 53 pregnant subject subjects. Second, 66 patients with ectopic pregnancy were compared with 51 pregnant control subjects. RESULTS: Geometric mean titers for Chlamydia trachomatis were higher among patients with tubal factor infertility and ectopic pregnancy, and they were more likely to have high antichlamydial titers (> or = 1:128 immunoglobulin G). Serum titer was significantly correlated with histologic evidence of salpingitis among the patients with an ectopic pregnancy. CONCLUSION: Our findings, similar to those from Western societies, suggest that among Egyptian women, prior chlamydial infection is associated with an increased risk of tubal factor infertility and possibly ectopic pregnancy.

PIP: This paper reports findings from a study which explored the association between past chlamydial infection, tubal factor infertility, and ectopic pregnancy in an Egyptian population. Conclusions are based upon findings from two concurrent case-control studies. In the first study, 51 patients with tubal factor infertility were compared with 48 healthy subjects who did not have tubal factor infertility and 53 pregnant subjects. In the second study, 66 patients with ectopic pregnancy were compared with 51 pregnant control subjects. Women who participated in the studies were patients of mean age approximately 29 years who presented for obstetrics and gynecology services at Shatby Hospital, Alexandria, Egypt, during 1989. The hospital serves a low- to lower-middle-class population. Geometric mean titers for Chlamydia trachomatis were higher among patients with tubal factor infertility and ectopic pregnancy, and they were more likely to have high antichlamydial titers. Further, serum titer was significantly correlated with histologic evidence of salpingitis among the patients with an ectopic pregnancy. These findings suggest that among Egyptian women, prior chlamydial infection is associated with an increased risk of tubal factor infertility and possibly ectopic pregnancy.

Chlamydial serologic characteristics among intrauterine contraceptive device users: does copper inhibit chlamydial infection in the female genital tract?

OBJECTIVE: Our objective was to assess the association between copper intrauterine contraceptive device use and the level of antichlamydial antibodies. STUDY DESIGN: Fifty-four women (29 with history of ectopic pregnancy, 2 with non-tubal factor infertility, 1 with tubal factor infertility, and 22 with intact intrauterine pregnancies) with current or prior intrauterine contraceptive device use were compared with 60 prenatal control subjects with respect to antichlamydial antibodies. RESULTS: When the intrauterine contraceptive device user group was subdivided into copper users and Lippes Loop device users, a significantly lower geometric mean serum antibody titer among copper intrauterine contraceptive device users was detected. The odds ratio estimates for past chlamydial exposure (> or = 1:64 immunoglobulin G, > or = 1:128 immunoglobulin G) in users of noncopper versus copper intrauterine contraceptive devices were 9.1 (95% confidence interval 1.9 to 43.0) and 10.5 (95% confidence interval 1.5 to 71.8), respectively. CONCLUSIONS: The lower geometric mean serum antibody titer of antichlamydial antibody among copper versus Lippes Loop device users and the large association measured between past chlamydial infection and noncopper intrauterine contraceptive device use suggest that copper may have a protective effect against Chlamydia trachomatis infection or a suppressive effect on development of antichlamydial antibodies.

PIP: In Alexandria, Egypt, clinicians compared data on 54 women who were either current or prior IUD users with data on 60 pregnant women who had never used any contraception to examine the association between IUD use and the level of antichlamydia antibodies. The IUD users had a history of ectopic pregnancy (29), intact intrauterine pregnancy (22), nontubal-factor infertility (2), and tubal factor infertility (1). The cases and controls had essentially the same geometric mean serum antibody titers for Chlamydia trachomatis (10.47 vs. 6.17). Yet, women who used copper-releasing IUDs had a lower geometric mean titer than did those who used the Lippes Loop IUD, but the difference was only significant for IUD only users and IUD users with a history of ectopic pregnancy. Specifically, among women who had used a copper IUD, the geometric mean serum antibody titers for those who only used 1 type of IUD, had used an IUD just for contraception, and those who had a history of ectopic pregnancy were 4, 5.9, and 3.8, respectively, while they were 24, 25.1 and 25.1, respectively, for their Lippes Loop counterparts (p 0.05). Among IUD users who had had an intrauterine pregnancy, the titer for women using a copper IUD was 5 compared to 14.8 for those using a Lippes Loop. When the researchers used a serum titer of 1:64 as a cutoff point for the 2 subgroups of IUD users, the Lippes Loop group had a higher proportion with titers above 1:64 than did the copper IUD group (47.6% vs. 9.09%; odds ratio [OR] = 9.09). Further, a higher proportion of Lippes Loop users had a titer greater than 1:128 than did copper IUD users (OR = 10.5). These results suggest that copper either inhibits the biologic effect of C. trachomatis or suppresses the immune response to chlamydia. Based on these findings and those of previous studies, the researchers believe the former is the most likely explanation.