Immunolocalization of Fertilin beta, IZUMO1, and P34H in Ram Spermatozoa

According to various reports, current methods of sperm freezing destroy the integrity of the sperm plasma membrane and acrosome. This study aimed to determine the changes in the existence and location of three proteins, namely fertilin beta, IZUMO1, and P34H, in ram spermatozoa. By using frozen-thawed spermatozoa, ejaculated fresh spermatozoa, and testicular and epididymal spermatozoa (obtained from caput, corpus, and caudal regions), the localizations of the mentioned proteins were performed using signal labeling with indirect immunofluorescence, and the quantification of these proteins was compared using Western blot analyses. Moreover, protein localization and signal labeling in fresh and frozen-thawed spermatozoa subjected to in vitro capacitation and acrosome reaction were compared. Using chlortetracycline (CTC) staining, as expected, it was detected that after incubating for 4 hours under capacitating conditions related to the control sample (0 hour), capacitated and acrosome-reacted sperm were increased (p < 0.001). Frozen-thawed samples had a lower density and expression than the ejaculate samples. Expression was not obtained, except for IZUMO1, from samples that underwent in vitro capacitation/acrosome reactions. Expression of IZUMO1 was seen as an increasing band formation from the equatorial region through the acrosome, after in vitro capacitation. However, after the acrosome reaction, the band formation was only on the equatorial region. Region-specific differences of proteins at the kDa level were obtained using Western blot analysis and possible isoforms specific to ram spermatozoa or proteins with similar epitopes were expressed. Considering the changes in surface proteins in frozen-thawed sperm, it is suggested that fertilin beta and P34H can be used as fertility or freezability markers

A comparative study of the effects of gutta-percha solvents on human osteoblasts and murine fibroblasts

We aimed to investigate the in vitro physiologic effects of xylene, chloroform, orange oil and eucalyptus oil solvents for dissolving gutta-percha on L929 and HOB cell lines; 2.5 and 10 mu L mL(-1) of these solvents were tested for 24, 48 and 72 h. Gutta-percha solvents inhibited the proliferation rate of fibroblasts in a dose- and time-dependent manner; however, no inhibition was detected in HOB (evaluated using MTT assay). None of the solvents induced apoptosis/necrosis in HOB cells at <= 2.5 mu L mL(-1) concentration in contrast to L929 (determined using acridine orange/ethidium bromide dual staining). Each solvent tested reduced the migration rate of both L929 and HOB cell lines in a dose-dependent manner (evaluated using a scratch assay). Gutta-percha solvents can damage fibroblast-rich tissues. Osteoblasts seemed to be more resistant to the tested solvents, and excessive extrusion of solvents from the root canal may also damage the periradicular tissues and reduce the ability to repair

Effects of Different Doses of Boric Acid Injected into Chicken Egg on Bursa of Fabricius and Spleen: A Histological and Stereological Study

The aim of the study was to investigate the effects of different doses of boric acid injected into chicken eggs on bursa of Fabricius and spleen. The study material consisted of three treatment and one control group. On the fourth day of incubation, boric acid dissolved in 0.9% NaCl were injected into eggs at 1.000, 1.500 and 2.000 ppm doses and control group received only 0.9% NaCl injection. Chicks hatched were raised until ten weeks of age (n= 10). Bursa of Fabricius and spleen were evaluated by histological, immunohistochemical and stereological methods. Weight of bursa of Fabricius, relative weight of bursa of Fabricius (P<0.01), volume of bursa of Fabricius, cortical thickness, follicle area and area of follicular medulla significantly reduced (P<0.001) by injection of 1.500 ppm boric acid compared to control group. The number of white blood cells also decreased in 1.000 ppm and 1.500 ppm groups. An increase (P<0.001) was recorded in hemoglobin ratio in 2.000 ppm group when compared with the other treatment groups and control group. There was an increase in 1500 ppm group in terms of spleen weight (P<0.001), relative spleen weight (P<0.01) and plasma cell count (P<0.05) and likewise an increase was noted in 1.000 and 2.000 ppm groups with respect to follicle count and number of apoptotic cells in comparison to control group. In conclusion, low doses of boric acid induced the bursal involution and implicitly increased plasma cell count in the spleen