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Introduction: Amenorrhea is a symptom of a variety of disorders and dysfunctions. Historically, there have been many practical difficulties associated with the diagnosis of amenorrhea due to the complex nature of the ovaries, pituitary gland and hypothalamus. Purpose: To develop a free, simple stand alone educational research software (ERS) to assist the education of clinicians and laboratorians (or undergraduate students) with regard to the clinicobiochemical evaluation of amenorrhea. Μaterials and methods: The software was designed using: a) Microsoft Windows as operating system, b) C# .NET (4.0) as software component (plug-in), and c) C# (C Sharp) as (object-oriented) programming language. It can be distributed on Compact Disk (CD) and be run on any Personal Computer (PC) on Windows. Results: The developed (ERS) -which we have called ERSA v.1.0 - does not require comprehensive skills and expertise in computers. Its educational benefits (common reasons for use) include activating motivation, stimulating recall of the prerequisite material, providing learning guidance and feedback (interactivity), usability outside timetabled course, and competency assessment. Conclusions: The free ERSA v.1.0 could be a practical digital teaching tool for supporting the clinicobiochemical education. Future research should continue so as to evaluate and improve its accuracy, appropriateness, and usability by healthcare students or professionals.
In the present study, the pattern of cyclooxygenase-2 (COX-2) expression in health and inflamed porcine uteri was analyzed using real-time reverse transcriptase-polymerase chain reaction (RT-PCR),Western blot and immunohistochemistry. On day 3 of the estrous cycle, 50 ml of saline or 50 ml of Escherichia coli (E. coli) suspension containing 10⁹ colony-forming units/ml, were injected into each uterine horn of the control (n=6) and experimental gilts (n=7), respectively. This latter procedure lead to a moderately (n=3) or severely intense (n=4) acute endometritis after eight days. Expression of both the COX-2 mRNA and protein was increased in the endometrium (ENDO) of animals suffering from the moderate (P<0.05, P<0.01, respectively) and severe (P<0.01) acute endometritis, as compared to the control tissues. Moreover, COX-2 mRNA level and protein content were higher (P<0.05) in the ENDO of animals with severe than with a moderately acute endometritis. An elevation in the COX-2 gene (P<0.05) and protein (P<0.001) expression was also observed in the myometrium (MYO) of animals suffering from severe endometritis, when compared with the levels observed in MYO of both the health and moderate intensely inflamed uteri. However, both the COX-2 mRNA and protein levels were similar in MYO of the control and moderately inflamed organs. The luminal epithelium, some of uterine glands and circular layer of the MYO were more intensely stained for COX-2 in animals with severe endometritis, than in animals with healthy or moderately inflamed uteri. Nonetheless, stronger COX-2 reaction was found in some of the uterine glands in latter group, when compared to that observed in uteri of the control animals. While positive COX-2-labeling was observed in the muscular layer of all arteries supplying the health and inflamed uteri, such staining was exclusively present in the endothelium of some arteries in inflamed organs. Likewise, some arteries in uteri of the animals with severe endometritis displayed immunoreaction stronger than that found in uteri of the animals with moderate inflammation. The present study revealed an up-regulation of COX-2 mRNA and protein in the inflamed porcine uterus, which was directly related to the intensity of the organ inflammation. An increase in the COX-2 expression in the uterus challenged by E. coli-induced inflammation indicates that this enzyme is crucial for elevated prostaglandins production in the inflamed organ.
The objective of this study was to determine the effect of long-term (48 d), per os animal administration of low zearalenone (ZEA) doses (50% and 100% NOAEL values) on the dynamics of changes in the morphometric parameters of the reproductive organs in sexually-immature gilts. The experiment involved 12 clinically-healthy gilts aged 2 months with initial body weight of ± 40 kg and a determined immune status. The animals were randomly divided into two experimental groups (El, n=4; E2, n=4) and a control group (C, n=4). Group El was administered per os 20 µg of ZEA/kg b.w. for 48 d, group E2 received per os 40 µg of ZEA/kg b.w. for 48 d, and group C was administered per os placebo for 48 d. The mycotoxin was administered daily per os animal in gelatin capsules before morning feeding. The animals were slaughtered at the end of the experiment. No significant morphometric changes were noted in the reproductive system of the gilts, except for an increase in the number of medium-sized ovarian follicles in group El. This suggests that ZEA at low concentrations may cause hormonal effects (hyperoestrogenism) but it does not exhibit xenobiotic activity.
The localisation of oestrogen (ER) and epidermal growth factor receptors (EGFR) in the various cell types of the bitch uterine was determined. In this study, 23 adult, healthy crossbred bitches brought to the clinic for ovariohisterectomy were used. ER and EGFR positive staining was detected in all cell types of the uterus. A distinct staining was seen in the luminal and glandular epithelium; while stromal and myometrial cells showed weak or moderate staining. The endothelial and smooth muscle cells of the vessels in the endometrium and myometrium sometimes appeared positive. No staining was observed in the mesothelium. The results of this study suggested that ER and EGFR were expressed at various levels in different cell types of bitch uterus. In light of the previous studies, and data of the presented investigations, it may be necesssary to elicit the harmonious proliferation and differentiation of epithelial and stromal cells that are considered essential for the preparation of the uterus for implantation.
Our previous studies suggested that acylation of the N-terminus of several known B2 antagonists with various kinds of bulky acyl groups consistently improved their antagonistic potency in rat blood pressure assay. On the other hand, our earlier observations also seemed to suggest that the effects of acylation on the contractility of isolated rat uterus depended substantially on the chemical character of the acyl group, as we observed that this modification might either change the range of antagonism or even transform it into agonism. Bearing all this in mind, we decided to synthesize seven new analogues of bradykinin by N-terminal acylation with various acyl groups of a moderately potent B2 antagonist, previously synthesized by Stewart's group, D-Arg-Arg-Pro-Hyp-Gly-Thr-Ser-D-Phe-Thi-Arg. The analogues were tested in vitro for their blood pressure-lowering and uterotonic activities. The modifications either preserved or increased the antagonistic potency in the rat blood pressure test. On the other hand, all seven substituents negatively influenced the interaction with the rat uterine receptors. Our results may be helpful for designing new B2 agonists and antagonists.
An eight-year-old German shepherd crossbreed clinically healthy bitch was ovariohysterectomized on the owner's request. The ovariohysterectomy operation was performed on the left flank area, under general anaesthesia. In both uterine horns, diffuse 0.1-3 cm in length cystic structures filled with serous fluid were detected during macroscopic examination. Histopathological examination of the uterus revealed that the pathological tissue was diffuse cystic lymphangiectasia.
Both, earlier investigations and complete formulas of pet food for dogs show that professionally prepared pet food is based in 30% on the material of the plant origin. Our own investigations prove that animal feeding stuffs of the plant origin are very often vectors of many undesirable substances (including micotoxins) that are unfavourable for carnivores. Zearalenone is a micotoxin, which frequently occurs among them. It was revealed in pet food in quantities from 5.0-299.5 μg/kg- It is a very dangerous xenobiotic because of its quick and easy absorption in the organism after per os application in monogastric animals. It is also a sterid and despite its plant origin it binds estrogen receptors in the reproductive tract and causes its hypo- and dysfunction. It is clearly visible in young, growing organisms and those during the menopause. In bitches, dysfunctions of the reproduction system, such as: endometritis-pyometra syndrome, prolonged oestrus, ovarian cysts and others are often found. It is supposed that specific for this species hormonal regulation of the reproduction processes based on the long progesterone and prolactin cycles, and on high sensibility to estrogens plays an important role in the etiopathogenesis of these dysfunctions. The application of hormones in bitches in therapeutic and biotechnical (contraception) purposes are the other factors, which should be taken into account in this aspect. There are some suggestions that therapeutic management, unfortunately connected with mistakes in the medical art, is often a cause of pathological condition in a reproductive organ in bitches. However, another reason of these disorders may be that bitches take zearalenone with the commercial pet food. The long-lasting intake of the feed with a low dose of this micotoxin may be the factor, which complicates hormonal regulation of the reproduction processes and is the cause of many disorders. Our own pilotage investigations showed that application of zearalenone at a dose of 200 μg/kg b. w. for 7 days leads to apoptotic-like changes in granule cells in the bitch reproductive system, atresia of follicular cells and oedema and hyperplasia of the oviductal and uterine cells. They could also indicate a follicular phase of the oestrus cycle in bitch. However, it did not take place.
The purpose of this study was to determine whether inflammatory changes in uterus caused by Escherichia coli are associated with changes in nitric oxide (NO) production. Therefore, the activity of NADPH-diaphorase (NADPH-d), a histochemical marker for nitric oxide synthase (NOS), localization of inducible isoform of NOS (iNOS) and tissue content of nitrite were studied in the uterine structures after inoculation of Escherichia coli into the uterus in gilts. Twelve sexually matured gilts with controlled estrous cycle were used. The animals were laparotomized on the 2nd day of the estrous cycle and polyvinyl cannulas were inserted into the uterine horns to infuse saline or Escherichia coli. In the group I (control; n=6), 25 ml of saline was infused into each uterine horn on the 4,h day of the estrous cycle. At the same time, 25 ml of Escherichia coli (strain 025:K23/α/:Hl) suspension, containing 107 colony forming units/ml was inoculated into each uterine horn of the treated gilts (group II; n=6). The middle part of the uterine horn was collected on the 14th day of the next estrous cycle immediately after slaughter. Cryostat sections from paraformaldehyde fixed tissues were stained histochemically to study the activity of NADPH-d and immunohistochemi- cally to investigate the distribution of iNOS. Optical density was assessed to evaluate the intensity of the histochemical reaction. Nitrite content was measured spectrophotometrically. In the Escherichia coli-treated gilts, the activitý of NADPH-d in the luminal epithelium and in external parts of excretory ducts of uterine glands was higher (P < 0.001) as compared to that in the control animals. In the secretory part of the uterine glands the activity of this enzyme was similar in both groups of the gilts. In the gilts that received Escherichia coli, the histochemical reaction of NADPH-d in endometrial blood vessels was stronger than that found in the control animals. Immunoreactivity for iNOS in the luminal epithelium, in external parts of excretory ducts of uterine glands and in vascular endothelial cells was stronger in the Escherichia coli-treated gilts as compared to that observed in the control animals. Only weak or no immunoreactivity was found in the secretory part of the uterine glands in both groups of the gilts. After Escherichia coli inoculation, nitrite content in the uterine tissues was higher (P < 0.05) than that determined in the controls. Our study has revealed that infusion of Escherichia coli into the porcine uterus induces the activity of NADPH-d, iNOS and increases the tissue content of nitrite in this organ. The data obtained indicate that NO can mediate an inflammatory effect of Escherichia coli in the uterus.
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