A comparative physicochemical and pharmacological evaluation of dexamethasone sodium phosphate and betamethasone sodium phosphate mucoadhesive gels for the treatment of oral submucous fibrosis in rats

Abstract The present study is aimed to formulate steroidal oral mucoadhesive gels of dexamethasone sodium phosphate and betamethasone sodium phosphate. Six gel formulations each of dexamethasone sodium phosphate and betamethasone sodium phosphate prepared using two different polymers carboxymethyl cellulose sodium and hydroxypropyl methylcellulose, in variable proportions. All the formulations subjected for assessment of various physicochemical parameters and mechanical properties. The formulations BSP5 and DSP5, both containing 1.25 % carboxymethyl cellulose sodium, 1.25 % hydroxypropyl methylcellulose, exhibiting mucoadhesive strength of 12.300 ± 0.004 and 12.600 ± 0.01, adhesiveness of 28.04 ± 00 and 30.02 ± 00, cohesiveness of 28.04 ± 00 and 30.02 ± 00, drug release of 86.869 ± 0.380 % and 88.473 ± 0.457 % respectively were considered as promising ones and were further subjected for stability studies and in vivo study in male albino rats. Formulation DSP5 upon oral application for 4 months in arecoline induced oral submucous fibrosis rats, showed more than 80 % reduction in fibrosis as compared with BSP5 which showed nearly 50 % reduction. These results were concluded on the basis of histopathological profile and weight gain among the experimental animals during in vivo study. Hence, DSP5 by minimizing the painful injuries and morbidities justifies being suitable noninvasive model for OSMF treatment.

Assessment of binder of sperm protein 1 (BSP1) and heparin effects on in vitro capacitation and fertilization of bovine ejaculated and epididymal sperm.

The present study evaluated the effect of binder of sperm protein 1 (BSP1) and/or heparin on in vitro bovine capacitation and fertilization rates using epididymal and ejaculated bovine sperm. Frozen-thawed sperm were selected and used in the following treatments. Control group: Fert-TALP medium without heparin; heparin (HEP) group: Fert-TALP with heparin (10 UI/ml); BSP1 group: Fert-TALP medium with BSP1 (10 µg/ml for ejaculated sperm; 40 µg/ml for epididymal sperm); HEP + BSP1 group: Fert-TALP medium with heparin (5 UI/ml) and BSP1 (5 µg/ml for ejaculated sperm; 20 µg/ml for epididymal sperm) and determined in vitro capacitation rates in different interval times (0, 15, 30 and 60 min) using the chlortetracycline fluorescence (CTC) method. Also, we evaluated the development rates of oocytes fertilized with ejaculated or epididymal sperm into the same treatments. Capacitation was greater and faster when ejaculated sperm were treated for 60 min with heparin compared with other treatments. However, developmental rates were similar in all treatments. For epididymal sperm, the treatments with BSP1 presented higher capacitation and fertilization rates compared with heparin (P < 0.05). The effects of heparin + BSP1 on capacitation and developmental rates did not cause any increase in capacitation or blastocyst rates compared with other groups for ejaculated or epididymal sperm. In conclusion, this study confirmed that either BSP1 and heparin can be used as capacitator agents for bovine ejaculated sperm during IVF. However, BSP1 seems to be more efficient compared with heparin for epididymal sperm. Furthermore, BSP1 and heparin have no synergic effects on sperm capacitation.

"Colonization routes and demographic history of Chirostoma humboldtianum in the central Mexican plateau".

In the present study we evaluate the population structure and potential colonization routes of the silverside Chirostoma humboldtianum through approximate Bayesian computations. Six microsatellite loci were amplified in a total of 288 individuals from six different locations covering the complete geographic distribution of the species. Additionally, two mitochondrial DNA markers, a D loop control region and cytochrome b were amplified in a subset of 107 individuals. The results found with microsatellites allow recovering well-structured populations that have experienced a drastic reduction in the effective population size. On the other hand, mtDNA sequences showed a moderate phylogeographic structure with shared haplotypes between geographic localities and signalsof a slight increase in the effective population size. Finally, the approximate Bayesian computation analysis performed with both datasets suggested a west-to-east colonization route for the species in Central Mexico.

Effect of the addition of sodium caseinate on the viability of cryopreserved buffalo semen / Efeito da adição de caseinato de sódio sobre a viabilidade do sêmen bubalino criopreservado

The use of cooled semen in artificial insemination operations results in higher pregnancy rates than the use of frozen semen. This result seems to be related to the more severe damage triggered by the freezing process than that observed during refrigeration. Due to its ability to bind to sperm-binding proteins and calcium ions, sodium caseinate has been studied as a substance capable of preventing early sperm capacitation, a significant cause of the decreased pregnancy rate resulting from the use of frozen semen. The first objective of this study was to evaluate whether a commercial egg yolk diluent developed for frozen bovine semen could be used for buffalo semen cryopreservation; the second objective was to investigate the effect of this diluent in combination with sodium caseinate during the procedures of buffalo sperm cryopreservation using flow cytometry and computer-assisted sperm analysis. In the first part of the study, comparing the results of spermatic kinetics and plasma and acrosomal membrane integrity, it was observed that the freezing process resulted in more cell damage than the cooling process. In the second part of the study, no effects of the addition of sodium caseinate to the egg yolk diluent were observed. From the results of the present study, it was possible to conclude that the egg yolk-based diluent was suitable for buffalo semen cryopreservation and that the addition of sodium caseinate did not decrease the harmful effects related to seminal cryopreservation.(AU)

O uso de sêmen resfriado em operações de inseminação artificial resulta em taxas de prenhez mais altas do que o uso de sêmen congelado. Esse resultado parece estar relacionado aos danos mais severos desencadeados pelo processo de congelação, quando comparado com o de refrigeração. Devido à sua capacidade de se ligar às proteínas ligadoras de espermatozoides e íons cálcio, o caseinato de sódio foi estudado quanto a sua capacidade de prevenir a capacitação espermática precoce, uma causa significativa de diminuição da taxa de prenhez com o uso de sêmen congelado. O primeiro objetivo deste estudo foi avaliar se um diluidor comercial a base de gema de ovo, destinado à congelação de sêmen bovino, poderia ser usado para a criopreservação de sêmen bubalino; o segundo objetivo foi investigar o efeito da adição do caseinato de sódio ao diluidor estudado durante os procedimentos de criopreservação de espermatozoides bubalinos. Foram empregadas a citometria de fluxo e a análise computadorizada do movimento espermático como métodos de avaliação seminal. Na primeira fase do estudo, comparando-se os resultados da cinética espermática e da integridade das membranas plasmática e acrossomal, observou-se que o processo de congelação promoveu mais danos celulares que o processo de resfriamento. Na segunda fase do estudo, não foram observados efeitos benéficos da adição de caseinato de sódio ao diluente empregado. A partir dos resultados do presente estudo, foi possível concluir que o diluente à base de gema de ovo foi adequado para a criopreservação do sêmen de bubalino e que a adição do caseinato de sódio não diminuiu os efeitos deletérios desencadeados pelo processo de criopreservação seminal.(AU)

Effect of the addition of sodium caseinate on the viability of cryopreserved buffalo semen / Efeito da adição de caseinato de sódio sobre a viabilidade do sêmen bubalino criopreservado

The use of cooled semen in artificial insemination operations results in higher pregnancy rates than the use of frozen semen. This result seems to be related to the more severe damage triggered by the freezing process than that observed during refrigeration. Due to its ability to bind to sperm-binding proteins and calcium ions, sodium caseinate has been studied as a substance capable of preventing early sperm capacitation, a significant cause of the decreased pregnancy rate resulting from the use of frozen semen. The first objective of this study was to evaluate whether a commercial egg yolk diluent developed for frozen bovine semen could be used for buffalo semen cryopreservation; the second objective was to investigate the effect of this diluent in combination with sodium caseinate during the procedures of buffalo sperm cryopreservation using flow cytometry and computer-assisted sperm analysis. In the first part of the study, comparing the results of spermatic kinetics and plasma and acrosomal membrane integrity, it was observed that the freezing process resulted in more cell damage than the cooling process. In the second part of the study, no effects of the addition of sodium caseinate to the egg yolk diluent were observed. From the results of the present study, it was possible to conclude that the egg yolk-based diluent was suitable for buffalo semen cryopreservation and that the addition of sodium caseinate did not decrease the harmful effects related to seminal cryopreservation.

O uso de sêmen resfriado em operações de inseminação artificial resulta em taxas de prenhez mais altas do que o uso de sêmen congelado. Esse resultado parece estar relacionado aos danos mais severos desencadeados pelo processo de congelação, quando comparado com o de refrigeração. Devido à sua capacidade de se ligar às proteínas ligadoras de espermatozoides e íons cálcio, o caseinato de sódio foi estudado quanto a sua capacidade de prevenir a capacitação espermática precoce, uma causa significativa de diminuição da taxa de prenhez com o uso de sêmen congelado. O primeiro objetivo deste estudo foi avaliar se um diluidor comercial a base de gema de ovo, destinado à congelação de sêmen bovino, poderia ser usado para a criopreservação de sêmen bubalino; o segundo objetivo foi investigar o efeito da adição do caseinato de sódio ao diluidor estudado durante os procedimentos de criopreservação de espermatozoides bubalinos. Foram empregadas a citometria de fluxo e a análise computadorizada do movimento espermático como métodos de avaliação seminal. Na primeira fase do estudo, comparando-se os resultados da cinética espermática e da integridade das membranas plasmática e acrossomal, observou-se que o processo de congelação promoveu mais danos celulares que o processo de resfriamento. Na segunda fase do estudo, não foram observados efeitos benéficos da adição de caseinato de sódio ao diluente empregado. A partir dos resultados do presente estudo, foi possível concluir que o diluente à base de gema de ovo foi adequado para a criopreservação do sêmen de bubalino e que a adição do caseinato de sódio não diminuiu os efeitos deletérios desencadeados pelo processo de criopreservação seminal.

Differential abundances of four forms of Binder of SPerm 1 in the seminal plasma of Bos taurus indicus bulls with different patterns of semen freezability.

The Binder of SPerm 1 (BSP1) protein is involved in the fertilization and semen cryopreservation processes and is described to be both beneficial and detrimental to sperm. Previously, the relationship of BSP1 with freezability events has not been completely understood. The objective of this work was to determine the differential abundance of the forms of the BSP1 protein in cryopreserved seminal plasma of Bos taurus indicus bulls with different patterns of semen freezability using proteomics. A wide cohort of adult bulls with high genetic value from an artificial insemination center was used as donors of high quality, fresh semen. Nine bulls presenting different patterns of semen freezability were selected. Two-dimensional gel electrophoresis showed differential abundance in a group of seven protein spots in the frozen/thawed seminal plasma from the bulls, ranging from 15 to 17 kDa, with pI values from 4.6 to 5.8. Four of these spots were confirmed to be BSP1 using mass spectrometry, proteomics, biochemical, and computational analysis (Tukey's test at P < 0.05). The protein spot weighing 15.52 ± 0.53 kDa with a pI value of 5.78 ± 0.12 is highlighted by its high abundance in bulls with low semen freezability and its absence in bulls presenting high semen freezability. This is the first report showing that more than two forms of BSP1 are found in the seminal plasma of Nelore adult bulls and not all animals have a similar abundance of each BSP1 form. Different BSP1 forms may be involved in different events of fertilization and the cryopreservation process.

Purification of binder of sperm protein 1 (BSP1) and its effects on bovine in vitro embryo development after fertilization with ejaculated and epididymal sperm.

The present study evaluated functional aspects of binder of sperm 1 (BSP1) in the bovine species. In a first experiment, cumulus-oocyte complexes (n = 1274) were incubated with frozen-thawed ejaculated sperm (18 hours) in Fert-TALP medium containing: heparin, 10, 20, or 40 µg/mL BSP1. Heparin followed by gelatin affinity chromatography was used for purification of BSP1 from bovine seminal vesicle fluid. With ejaculated sperm, cleavage rates were similar when Fert-TALP medium was incubated with heparin (74.1 ± 2.7%), 10 µg/mL BSP1 (77.8 ± 3.1%), or 20 µg/mL BSP1 (74 ± 2.0%). Day-7 blastocyst rates were equivalent after incubations with heparin (40.8 ± 5.0%) and 10 µg/mL BSP1 (34.1 ± 4.4%), but reduced after 20 µg/mL BSP1 (22.4 ± 2.9%) and 40 µg/mL BSP1 (19.3 ± 4.1%; P < 0.05). In the second experiment, cumulus-oocyte complexes (n = 1213) were incubated with frozen-thawed cauda epididymal sperm (18 hours) in Fert-TALP medium containing: no heparin, heparin, 10, 20, or 40 µg/mL. Cleavage and blastocyst rates were similar after treatments with heparin (68.5 ± 1.3% and 24.7 ± 3.2%, respectively) or without heparin (65.5 ± 1.8% and 27.3 ± 1.6%, respectively). Cleavage was higher after treatment with any BSP1 concentrations (74.2 ± 2.7%-79.0 ± 1.1%) than without heparin (P < 0.05). Also, cleavage was better after Fert-TALP medium incubation with 40 µg/mL BSP1 (79.0 ± 1.1%) than with heparin (68.5 ± 1.3%; P < 0.05). Embryo development was higher (P < 0.05) after treatment with 20 µg/mL BSP1 (35.6 ± 2.5%) and 40 µg/mL (41.1 ± 2%) than after incubations with heparin (24.7 ± 3.2%) or without heparin (27.3 ± 1.6%). Interestingly, BSP1 did not cause reductions in blastocyst rates after fertilization with epididymal sperm, as observed with ejaculated sperm. On the basis of immunocytochemistry, there was BSP1 binding to frozen-thawed ejaculated but not to epididymal sperm. Also, anti-BSP1 reaction remained on ejaculated sperm (as expected) and appeared on epididymal sperm after incubation with purified BSP1. Acrosome reaction of ejaculated and epididymal sperm was induced after incubation with purified BSP1 as well, indicating an effect of BSP1 on capacitation. In conclusion, purified BSP1 from bull seminal vesicles was able to bind to and induce capacitation of ejaculated and epididymal sperm. Also, BSP1 added to fertilization media and allowed proper cleavage and embryo development, with the effects being modulated by previous exposure or not of spermatozoa to seminal plasma.

IGF-I, leptina, insulina e proteínas associadas à qualidade do plasma seminal: ação local / IGF-I, leptine, insulin and proteins associated to seminal plasma quality: local action

É possível que a expressividade de alguns elementos do plasma seminal dos bovinos, como proteínas e hormônios, possa servir como marcadores para sêmen de alta ou baixa fertilidade. Vários estudos têm demonstrado a associação de proteínas do plasma seminal com a fertilidade de touros. Dentre as mais estudadas, destacam-se aquelas com afinidade à heparina, que exercem importantes papéis na capacitação espermática e na reação acrossômica. Alguns fatores endócrinos e/ou locais, podem estar associados à expressividade e/ou função destas proteínas, auxiliando nas condições espermáticas favoráveis à fecundação. Dentre estes, destacam-se a insulina, a leptina e o fator de crescimento semelhante à insulina do tipo I. Assim sendo, evidenciam diferenças entre animais, estando associados à estrutura e as condições metabólicas da célula espermática, auxiliando na determinação da qualidade do plasma seminal. Desta maneira, o estudo das proteínas do plasma seminal asociado à condição metabólica destes hormônios, presentes neste meio, pode servir como importante parâmetro de avaliação da condição reprodutiva do macho

It is possible that the expression of some elements in seminal plasma, such as proteins and hormones, may act as markers of quality. Several studies have demonstrated an association of seminal plasma proteins with fertility. Among the most studied, there are the proteins with affinity to heparin, which play an important role in sperm capacitation and acrosome reaction. Some endocrine factors and/or locals may be associated with the expression and function of these proteins, aiding in sperm condition conducive to fertilization. Among them stand out as insulin, leptin and growth factor insulin-like type I. Thesefactors may highlight the differences between these animals because they are associated with the metabolic condition and structure of sperm cells, aiding in determining the quality of seminal plasma. Thus, the study of seminal plasma proteins associated with the metabolic condition of these hormones are present in this medium can serve as a new parameter for assessment of male reproductive condition...