-
Asian-Australasian Journal of Animal... Jan 2018In this study, we investigated the adverse effects of dietary zearalenone (ZEA) (0.5 to 1.5 mg/kg diet) on the localization and expression of the growth hormone receptor...
OBJECTIVE
In this study, we investigated the adverse effects of dietary zearalenone (ZEA) (0.5 to 1.5 mg/kg diet) on the localization and expression of the growth hormone receptor (GHR) in the uteri of post-weaning gilts and explored alternative mechanism of the reproductive toxicity of ZEA on piglets.
METHODS
A total of forty healthy piglets (Duroc×Landrace×Large White) aged 28 d were selected for study. Piglets were transferred to single cages after 10 days' adaptation on an obstetric table. The animals were allocated to one of four treatments: a normal basal diet supplemented with 0 (Control), 0.5 (ZEA0.5), 1.0 (ZEA1.0), or 1.5 (ZEA1.5) mg/kg purified ZEA, and fed for 35 d after the 10-d adaptation. Analyzed ZEA concentrations in the diets were 0, 0.52±0.07, 1.04±0.03, and 1.51±0.13 mg/kg, respectively. At the end of the feeding trial, piglets were euthanized after being fasted for 12 h. Two samples of uterine tissue from each pig were rapidly collected, one of which was stored at -80°C for analysis of the relative mRNA and protein expression of GHR, and the second was promptly fixed in Bouin's solution for immunohistochemical analysis.
RESULTS
The relative weight of the uteri and thickness of the myometrium and endometrium increased linearly (p<0.001) and quadratically (p<0.001) with an increasing level of ZEA. The results of immunohistochemical analysis indicated that GHR immunoreactive substance was mainly localizated in the cytoplasm of uterine smooth muscle, glandular epithelial, luminal epithelial, stromal, and vascular endothelial cells. In contrast, nuclear staining was rarely observed. The immunoreactive integrated optic density of GHR in the myometrium, luminal epithelium, glandular epithelium, and whole uteri of weaning gilts increased linearly (p<0.001) and quadratically (p<0.05) with an increasing level of ZEA. The mRNA and protein expression of GHR in the uteri of weaning gilts increased linearly (p<0.001) and quadratically (p<0.05) with an increasing level of ZEA.
CONCLUSION
In conclusion, ZEA at a concentration of 0.5 mg/kg was sufficient to significantly thicken the myometrium and endometrium, and at a concentration of 1.0 mg/kg induced a high level of GHR expression to promote growth and development of the uteri. This revealed an alternative molecular mechanism whereby ZEA induces growth and development of the uteri and provides a theoretical basis for the revision of Chinese feed hygiene standards.
PubMed: 28920404
DOI: 10.5713/ajas.17.0526 -
JBRA Assisted Reproduction Oct 2021This comparative study was designed to analyze the potential effects of Yaji (suya meat sauce) and its composite spices on male fertility based on testicular histology,...
OBJECTIVE
This comparative study was designed to analyze the potential effects of Yaji (suya meat sauce) and its composite spices on male fertility based on testicular histology, serum testosterone level, and semen analysis parameters.
METHODS
The study included 70 adult male Sprague Dawley rats with an average weight of 120 g. They were divided into two experimental study groups, respectively analyzed for 28 and 56 days. Each group featured 35 rats, further subdivided into seven treatment groups (A - G; n=5 each). Group A - Control; Group B: 200 mg/kg of Yaji; Group C: 200 mg/kg of red pepper; Group D: 200 mg/kg of black pepper; Group E: 200 mg/kg of clove; Group F: 200 mg/kg of ginger; and Group G: 200 mg/kg of garlic given orally using an oral cannula. At the end of the experiment, the animals were euthanized. Blood samples collected via cardiac puncture and their testes were excised and weighed. The cauda epididymis was excised for semen analysis using a Neubauer Counting Chamber (hemocytometer) and the testes were fixed in Bouin solution, processed, and stained with Hematoxylin and Eosin.
RESULTS
Significant increases (p<0.05) were seen in body weight, testicular weight, serum testosterone level, sperm count and motility in the Yaji treated groups, in addition to significant increases in serum testosterone level, sperm counts, and sperm motility, and enhanced spermatogenesis and proliferation of Leydig cells in vivo as compared to the groups given isolated component spices (groups C-G), which also showed significant changes in testosterone and semen analysis when compared with the control groups.
CONCLUSIONS
Yaji or its spice components can boost male fertility parameters when consumed in moderated quantities without the known cytotoxic additives or condiments such as monosodium glutamate.
Topics: Animals; Male; Meat; Rats; Rats, Sprague-Dawley; Sperm Count; Sperm Motility; Spermatozoa; Spices
PubMed: 34224239
DOI: 10.5935/1518-0557.20210020 -
Folia Histochemica Et Cytobiologica 2007Apoptosis is a natural process which accompanies human ovary from the moment of birth until old age. While it is a well-known process at the reproductive age, it still...
Apoptosis is a natural process which accompanies human ovary from the moment of birth until old age. While it is a well-known process at the reproductive age, it still needs to be thoroughly examined when referring to the postmenopausal age. The study involved 30 postmenopausal women who had their ovaries removed by laparotomy due to nonneoplastic diseases of the uterus. The women were divided into 3 groups depending on the time that had passed since the last menstruation. Group A consisted of women who had their last menstruation no more than 5 years earlier. In group B menopause occurred 5 to 10 years earlier. Group C was composed of patients who had the last menstruation over 10 years earlier. In all the patients concentrations of follitropin (FSH) and estradiol (E2) in blood plasma were measured. Ovarian tissue was obtained during surgery. For morphological studies, ovaries were fixed in Bouin's solution and 4% formalin and embedded in paraffin. Morphological analysis was carried out after hematoxylin-eosin (H-E) staining. For histochemical detection of apoptotic cells (in situ localization of fragment DNA), the TUNEL method was used. The expression of caspase-3 positive cells was determined immunohistochemically in paraffin-embedded specimens. Comparing to groups A and B, the ovaries in group C contained small number of corpora albicantia located in the medullary part as well as thinned blood vessels and few lymphatic vessels and nerves. In contrast to group A where the number of TUNEL-positive cells was high and caspase-3 expression was observed, no TUNEL-positive nuclei and caspase-3 expression were found in the examined ovaries of group C women.
Topics: Apoptosis; Caspase 3; Estradiol; Female; Follicle Stimulating Hormone; Humans; Immunohistochemistry; In Situ Nick-End Labeling; Middle Aged; Ovary; Postmenopause
PubMed: 17597023
DOI: No ID Found -
The Journal of Cell Biology Jul 1962The ultrastructure of the uterine epithelium has been studied in estrous, ovariectomized, pregnant, and pseudopregnant rabbits. Tissue for light microscopy was fixed in...
The ultrastructure of the uterine epithelium has been studied in estrous, ovariectomized, pregnant, and pseudopregnant rabbits. Tissue for light microscopy was fixed in Bouin's solution and stained with hematoxylin and eosin, by the periodic acid-Schiff (PAS) method, and with methylene blue. Tissue for electron microscopy was fixed in 1 per cent osmium tetroxide in White's saline and embedded in Araldite. The uterine epithelium in estrus is comprised of ciliated and non-ciliated cells. After ovariectomy the epithelium becomes reduced in height and PAS-positive material disappears. Multinucleated cells are formed in the epithelium in pregnancy, pseudopregnancy, and in the non-pregnant horn in unilateral pregnancy. They degenerate during the 3rd week of pseudopregnancy and during the 4th week of pregnancy in the non-pregnant horn. The formation of multinucleated cells is believed to be under hormonal control. The uterine epithelium in contact with the blastocyst changes into a "symplasma," presumably under the influence of a local (chemical?) effect produced by the blastocyst. This change is not seen in pseudopregnancy nor in the non-pregnant horn in unilateral pregnancy. A complex infolding of the basal cell membrane of the epithelium accompanies the "symplasmic" change. The remaining uterine epithelium in pregnancy shows a well developed ergastoplasm suggesting a production of secretion materials, some of which may be available for absorption by the fetus through the yolk sac or paraplacental chorion.
Topics: Animals; Blastocyst; Cell Membrane; Electrons; Epithelium; Estrus; Female; Humans; Microscopy; Microscopy, Electron; Pregnancy; Pseudopregnancy; Rabbits; Uterus
PubMed: 14462496
DOI: 10.1083/jcb.14.1.49 -
Acta Cirurgica Brasileira Dec 2013To evaluate the effects of copaiba oil ointment (Copaifera langsdorffii) on dorsal skin flaps in rats.
PURPOSE
To evaluate the effects of copaiba oil ointment (Copaifera langsdorffii) on dorsal skin flaps in rats.
METHODS
Adult male rats (n=30) were distributed into three groups of ten animals each, as follows: GC--control; GCA--absolute control and GT--treated with copaiba ointment. The rats were subjected to dorsal cutaneous skin flap surgery and the animals from the GC and GT received post-operative treatment for eight consecutive days. The animals from the GCA group did not receive treatment while the animals from the GC group received daily topical treatment of ointment without the active ingredient and the animals from the GT group were daily treated with 10% copaiba oil ointment. At the end of each experimental period the lesions were evaluated according to the percentage of necrotic area. Then, fragments from cranial, median and caudal parts were fixed in Boüin's solution and processed for paraffin embedding. The morphology of histological sections (5µm) was evaluated and the number of leucocytes, fibroblasts and blood vessels was also analyzed. The data obtained were submitted to ANOVA test complemented by Tukey-Kramer test (p<0.05).
RESULTS
The necrotic area was lower in the group treated with copaiba ointment when compared to the control groups (GCA>GC and GT), while the morphology showed larger granulation tissue with bulky fibroblasts and collagen fibers more arranged in the GT group. The morphometry showed a significant higher number of blood vessels in the median and caudal parts (GT>GCA and GC), leucocytes in the cranial part (GT>GC>GCA), and also fibroblasts in the median (GT and GC> GCA) and caudal parts (GT>GC and GCA) (p<0.05).
CONCLUSION
The copaiba oil ointment favors angiogenesis and accelerates the viability of random skin flaps in rats.
Topics: Administration, Topical; Animals; Blood Vessels; Fabaceae; Fibroblasts; Leukocyte Count; Male; Ointments; Plant Oils; Random Allocation; Rats; Reproducibility of Results; Skin; Surgical Flaps; Tissue Survival
PubMed: 24316860
DOI: 10.1590/s0102-86502013001200009 -
The Journal of Parasitology Feb 1991Fresh (36 days old) sporulated oocysts of Eimeria nieschulzi were divided into 7 groups. Control oocysts were maintained at 23 C in 2% aqueous (w/v) K2Cr2O7. The 6...
Fresh (36 days old) sporulated oocysts of Eimeria nieschulzi were divided into 7 groups. Control oocysts were maintained at 23 C in 2% aqueous (w/v) K2Cr2O7. The 6 experimental groups were mixed with either Bouin's solution, 10% aqueous (v/v) buffered formalin, Karnovsky's solution, glutaraldehyde, paraformaldehyde, or 70% aqueous (v/v) ethanol (EtOH). After 115 days, oocysts from all 7 groups were examined under oil immersion to determine the effect of fixation on their structural integrity. The parameters examined were lengths and widths of oocysts and sporocysts, percent sporulation (%S), and percent crenation (%C) of oocysts and sporocysts. The highest destruction (%S and %C) occurred in oocysts exposed to glutaraldehyde and Karnovsky's fixatives where 100% of both oocysts and sporocysts crenated and only 8% and 48%, respectively, remained sporulated. Of the oocysts in paraformaldehyde, 93% remained sporulated, but 95%of these oocysts and 100% of the sporocyst crenated. In Bouin's solution, 75% of the oocysts were intact structurally, but of these, only 60% were still sporulated with 70% of their sporocysts crenated. Oocysts preserved in 70% EtOH were 80% intact and 70% remained sporulated, but nearly 60% of their sporocysts collapsed even though the oocyst walls were intact. Oocysts preserved in 10% buffered formalin maintained structural integrity but had lower numbers of sporulated oocysts (84%) and greater numbers of crenated oocysts (18%) than control oocysts maintained in the dichromate solution (95% and 0%, respectively).
Topics: Acetates; Acetic Acid; Animals; Eimeria; Ethanol; Fixatives; Formaldehyde; Glutaral; Picrates; Polymers; Preservation, Biological
PubMed: 1899451
DOI: No ID Found -
Veterinary Microbiology Sep 1994Feline coronavirus infections in cell cultures and in fresh and fixed feline tissues were detected using a polymerase chain reaction (PCR) test. Cell cultures were...
Feline coronavirus infections in cell cultures and in fresh and fixed feline tissues were detected using a polymerase chain reaction (PCR) test. Cell cultures were inoculated with feline infectious peritonitis virus (FIPV), feline enteric coronavirus (FECV) or sham inoculum. The tissue samples of liver, kidney and spleen were taken from specific-pathogen-free (SPF) cats that were inoculated intranasally with 10(3) TCID50 of FIPV 79-1146 (n = 10), FIPV UCD1 (n = 3) or sham inoculum (n = 3), from clinical cats (n = 43), and from formalin-fixed archived feline tissues (n = 49), respectively. Additional tissue samples were taken from the FIPV-inoculated cats (n = 6) and were kept at 4 degrees C, room temperatures (20-24 degrees C) and 37 degrees C respectively for 0, 6, 12, 24, 48, 72, and 96 hours before frozen (-70 degrees C) for PCR to evaluate the effects of the ambient temperatures and post-mortem intervals on the test. The samples were also fixed in 10% neutrally buffered formalin, 95% ethanol, and Bouin's solution respectively to evaluate the effects of the fixatives on the test. Positive PCR results were obtained from the cell cultures that were inoculated with FIPV and FECV and from the FIPV-inoculated cats (13/13). Negative PCR results were obtained from the sham-inoculated cell cultures and cats (3/3). Of the 92 clinical cats, 7 of the 8 FIP-suspected cats (87.5%) and 51 of the 84 non-FIP-suspected cats (60.7%) were shown to be virus-positive in at least one of the tissue samples. There was no significant difference in the PCR results between the fresh and the formalin-fixed tissues of the clinical cats (P > 0.05). Of the FIPV inoculated cats, the virus was detectable equally well in fresh and formalin-, Bouin's solution- or ethanol-fixed tissues. However, the amounts of total RNA extracted from the fixed tissues were significantly less than those from fresh tissues (P < 0.01). In tissues that were kept at 4 degrees C, the virus was detectable up to 96 h; at room temperatures, up to 48 h; and at 37 degrees C, up to 24 h, respectively.
Topics: Animals; Base Sequence; Cat Diseases; Cats; Cells, Cultured; Coronavirus; Coronavirus Infections; DNA, Viral; Molecular Sequence Data; Nucleic Acid Hybridization; Polymerase Chain Reaction; RNA, Viral
PubMed: 7839586
DOI: 10.1016/0378-1135(94)90078-7 -
Animals : An Open Access Journal From... Mar 2024Fixatives and fixation protocol have a profound effect on both the morphology and epitope sensitivity of ovarian tissue, which hampers accurate ovarian tissue...
Fixatives and fixation protocol have a profound effect on both the morphology and epitope sensitivity of ovarian tissue, which hampers accurate ovarian tissue evaluation. We aimed to establish the most suitable fixation protocol for feline () ovarian tissue. Fragments (1.5 mm diameter) were punched from 1 mm-thick feline ovarian tissue, divided into three groups then fixed with three different fixatives (Bouin, neutral buffered formalin [NBF] and form acetic acid [new compound fixative formulation for ovarian tissue composed of 5% acetic acid in NBF]) for five fixation periods. Subsequently, fragments were processed and evaluated for the morphology and intensity of immunohistochemical signals against three antigens (Ki-67, MCM-7 and activated caspase-3). Proportions of grade 1 or morphologically intact follicles were significantly lower in NBF when compared with Bouin and form acetic acid fixatives. However, Bouin fixative had the lowest mean DAB intensity ( < 0.05) in all three antigen targets, while NBF had the highest ( < 0.05) in Ki-67 and caspase-3, but in MCM-7, it was no different from form acetic acid. In conclusion, form acetic acid maintained ovarian tissue architecture with excellent follicular morphology in the same manner as Bouin fixative, and it also maintained reasonable DAB signals similar to NBF, thus providing a better alternative for feline ovarian tissue studies.
PubMed: 38539923
DOI: 10.3390/ani14060825 -
Journal of Toxicologic Pathology Jul 2018To prevent fixation defects or artifacts in the whole bodies of fish caused by conventional fixatives, such as formalin solution, Bouin's fluid (BF), and Davidson's...
To prevent fixation defects or artifacts in the whole bodies of fish caused by conventional fixatives, such as formalin solution, Bouin's fluid (BF), and Davidson's fluid (DF), the optimal fixatives and fixing method were examined. An improved method of fixing the whole bodies of fish was examined that makes use of a combination of 20% formalin and BF or DF. The fixatives were examined with four representative tissues, i.e., the gill, liver, intestinal tract, and kidney, to evaluate end points including the appearance of degraded tissues and artifacts caused by each fixative, overall morphological clarity of nuclei, staining intensity, and integrity of the other tissues. The best results were obtained when the fresh whole bodies were initially fixed in 20% formalin (primary fixation) at 4°C for 1 h and subsequently fixed in BF for 5 h at 4°C (secondary fixation). Therefore, the current findings led the authors to conclude that the combination of primary fixation with 20% formalin at 4°C for 1 h and secondary fixation with BF at 4°C for 5 h was suitable for fixation of the whole bodies of fish.
PubMed: 30093790
DOI: 10.1293/tox.2018-0001 -
Iranian Journal of Basic Medical... Jan 2018Lipopolysaccharide (LPS)-induced endotoxemia is known to cause male infertility. This study was designed to explore the effects of bacterial LPS on histomorphometric...
OBJECTIVES
Lipopolysaccharide (LPS)-induced endotoxemia is known to cause male infertility. This study was designed to explore the effects of bacterial LPS on histomorphometric changes of mice testicular tissues.
MATERIALS AND METHODS
In experiment 1, a pilot dose responsive study was performed with mice that were divided into five groups, receiving 36000, 18000, 9000, and 6750 µg/kg body weight (B.W) of LPS or only saline (control). White blood cells (WBC) were observed for 3 days after LPS inoculation. In experiment 2, two groups of mice were treated with 6750 µg/kg B.W of LPS or only saline (control). Five cases from each experimental group were sacrificed at 3, 30, and 60 days after LPS inoculation. Left testes were fixed in Bouin's solution, and stained for morphometrical assays.
RESULTS
Time-course changes of WBC obtained from different doses of LPS-treated mice showed that inoculation of 6750 µg/kg B.W produced a reversible endotoxemia that lasts for 72 hr and so it was used in the second experiment. In experiment 2, during the first 3 days, no significant changes were observed in the evaluated parameters instead of seminiferous tubules diameter. Spermatogenesis, Johnsen's score, meiotic index, and epithelial height were significantly affected at 30 day. However, complete recovery was only observed for the spermatogenesis at day 60. Interestingly, deleterious effects of LPS on spermatogonia were only seen at 60 day (<0.05).
CONCLUSION
Endotoxemia induced by LPS has long-term detrimental effects on spermatogonia and later stage germ cells, which are reversible at the next spermatogenic cycle.
PubMed: 29372036
DOI: 10.22038/IJBMS.2017.24415.6083