The frontal planar QRS/T angle in patients with non-ischemic type late gadolinium enhancement on cardiac magnetic resonance.

OBJECTIVE: Evaluation of myocardial fibrosis may offer an insight into underlying pathological mechanisms of ventricular arrhythmias. We intended to investigate the possible association between the presence and extent of myocardial fibrosis and ventricular repolarization heterogeneity based on frontal planar QRS/T angle. PATIENTS AND METHODS: We retrospectively investigated patients in whom gadolinium-based contrast agent cardiac magnetic resonance imaging was performed due to the suspicion of any myocardial disease. Patients with non-ischemic type late gadolinium enhancement (LGE) were enrolled into this study. The association between presence and extent of myocardial LGE and frontal planar QRS/T angle defined as the absolute difference between QRS wave axis and T-wave axis on a resting 12-lead surface ECG was evaluated. RESULTS: The frontal planar QRS/T angle was significantly higher in patients with myocardial fibrosis indicated by LGE compared to those without LGE (61.67 ± 40.70 vs. 37.27 ± 32.35, p < 0.001). LGE extent score assessed by visual 17-segment model was the only independent variable, which had a significant effect on frontal planar QRS/T angle [Unstandardized Coefficients B = 4.052, 95% CI [(2.025) - (6.079), p < 0.001]. CONCLUSIONS: In conclusion, this study showed that inhomogeneous areas of myocardium due to varying degrees of myocardial fibrosis might affect the electrical activity of the left ventricle, even with normal left ventricular dimensions and function.

Immunohistochemical detection of insulin-like growth factor-I, transforming growth factor-beta2, basic fibroblast growth factor and epidermal growth factor-receptor expression in developing rat ovary.

The aim of this study was to determine the immunohistochemical expression and localization of insulin-like growth factor-I (IGF-I), transforming growth factor-beta2 (TGF-beta2), basic fibroblast growth factor (bFGF) and epidermal growth factor-receptor (EGF-R) in developing rat ovaries. Eighteen female Wistar rats were enrolled in this study; newborn (n=6), one-month-old (n=6) and adult (n=6) rats. Formalin-fixed and parafin-embedded ovarian tissues were stained with antibodies against IGF-I, TGF-beta2, bFGF and EGF-R, immunohistochemically. The ovarian cells were evaluated by semi-quantitative scoring system under light microscope. The staining of IGF-I, TGF-beta2, bFGF and EGF-R were most intense in the oocytes and were heavily at one-month-old rats. A moderate immunostaining in theca cells and corpus luteii reacted with IGF-I in adult rats. Furthermore the staining intensity for IGF-I was moderate in granulosa cells of newborn rat ovaries. We detected also a moderate staining for TGF-beta2 in corpus luteii of adult rats. In addition, we found a bFGF immunostaining mainly in oocytes of follicles of young and adult rats. Immunostaining for EGF-R was moderate in granulosa cells of one-month-old rats. In conclusion, this study suggests that growth factors play a pivotal role in ovarian function, especially in follicular development. The role of growth factor in controlling degeneration or growth (or both) of ovary follicles remain as explained.

Expression and localisation of Bcl-2 and Bax proteins in developing rat ovary.

Bcl-2 and Bax proteins localised mainly in granulosa cells. Primordial and primary follicles of new born rat ovary showed an intensive nuclear staining for Bax but faint staining for Bcl-2. In terms of staining intensity, no remarkable difference was observed within the same stage of developing follicle. Compared to new born rats, granulosa cells of adult and one month old rat ovary showed an increased staining both for Bcl-2 and Bax proteins. No staining was observed in primordial follicles of one month old and adult rats. However, granulosa cells of primary follicles, granulosa cells and theca cells in tertiary follicles of adult rat ovary also showed a strong staining for Bcl-2 and Bax proteins. Oocytes of follicles from different developmental stages revealed an apparent staining both for Bcl-2 and Bax proteins. However, in the more mature follicles oocytes stained more intensively. In developing corpus luteum a remarkable staining was observed for Bax. However, the staining was more prominent in regressing corpus luteum. Contrary to this, Bcl-2 stained the luteal cells in developing corpus luteum strongly, while in the fully developed corpus luteum no staining for Bcl-2 was observed. In conclusion, there was an apparent relation between the expression of the apoptosis regulating protein Bcl-2 and Bax and follicular development. Thus, during the follicular development Bcl-2 and Bax may be involved in granulosa cell demise in rat ovary. Furthermore, increased levels of Bax and decreased levels of Bcl-2 in the fully developed corpus luteum suggest that Bax plays a role in apoptosis of luteal cells in rat ovary.

Early maternal changes contributing to the formation of the chorioallantoic and yolk sac placentas in rat: a morphological study.

In order to determine the maternal changes contributing to the formation of the chorioallantoic and yolk-sac placentas, rat gestation sites were examined by light and electron microscopy on days 7 through 10 of pregnancy. On day 7, the implantation chamber showed different compartments and contained the blastocyst in the antimesometrial chamber. The epithelial lining of the implantation chamber disappeared at the antimesometrial chamber, transformed into disintegrated cells in the mesometrial chamber, and showed signs of the programmed cell death in the decidual crypt. On day 8, the mesometrial chamber lumen contained red blood cells and it was continuous with subepithelial sinusoids. The endothelial cells lining the mesometrial sinusoids also showed some characteristics of the sprouting type angiogenesis such as hypertrophy and cell proliferation. While the yolk-sac placental circulation was more obvious with participation of the giant trophoblasts at the antimesometrial pole of the conceptus on day 9, the antimesometrial cells showed autophagic degeneration after the formation of the chorioallantoic placenta on day 10. The contribution of the regional cell death and angiogenesis to form both of the two placentas are discussed.

Heat shock preconditioning and pretreatment with glucocorticoid antagonist RU 486 protect rat myogenic cells H9c2 against glutamate-induced cell death.

We have observed that the treatment of rat-heart derived H9c2 myoblasts for 20 h with the excitatory amino acid glutamate resulted in cell death in a dose dependent manner as determined by LDH release. The optimum cardiotoxicity was seen at 25 mM glutamate. Preconditioning with either sublethal heat shock (42 degrees C for 30 min) or pretreatment with 500 nM of the glucocorticoid antagonist RU 486 for 24 h almost completely protected H9c2 cells against subsequent 20 h treatment with 25 mM lethal glutamate. In addition, we have observed that glutamate treatment resulted in intense nuclear localization of glucocorticoid receptors (GR) in H9c2 cells as judged by the confocal immunofluorescence microscopy. Furthermore, pretreatment with either heat shock or RU 486 followed by glutamate treatment resulted in dramatic decrease in GR nuclear localization which was almost comparable to that observed with control untreated cells. In conclusion, we have shown for the first time using H9c2 cells that (i) protection from glutamate cardiotoxicity occurs with prior treatment with sub lethal heat shock or RU 486 and (ii) these measures down regulate the intense nuclear localization of GR induced by glutamate. The block to GR nuclear localization is likely to be involved in cardioprotective effects offered against glutamate toxicity by pretreatment with heat shock or RU 486.

Biological effects of long-term caloric restriction: adaptation with simultaneous administration of caloric stress plus repeated immobilization stress in rats.

In the present study, we have established the biological effects during 8 weeks of (i) caloric restriction (Cal) and (ii) simultaneous administration of Cal plus 2 hr daily immobilization stress using male Sprague-Dawley rats. Animals were divided into three equal groups: (i) ad libitum fed, (ii) 30% restriction of food intake of the ad libitum diet, and (iii) 30% restriction of food intake plus 2 hr daily immobilization stress. Caloric-restricted animals gained only 30% of the total body weight of the unrestricted animals but received 70% of the food of those rats. Cal animals showed a significant loss in their relative liver and-thymus weight and a significant gain in their relative adrenal and testis weight as compared to the control animals. Cal animals had almost 2-fold higher levels of plasma corticosterone levels with a dramatic decrease in the total glucocorticoid receptor (GR) levels in the liver, thymus, heart, and testis as compared to ad libitum fed control animals. Interestingly, Cal animals showed higher levels of lipid peroxidation in both the liver and heart, indicating increased oxidative activities in these tissues when compared with the control animals. In addition, Cal animals had increased heat shock protein 70 (HSP 70) content in the testis. Surprisingly, hardly any significant differences were observed in either total body weight gain, organ weights, plasma corticosterone levels, or lipid peroxidation between Cal animals and Cal plus immobilization-stressed animals. The results obtained suggest that (i) several stress-related responses such as inhibition of total body weight gain, increased adrenal weight, decreased thymus weight, increased plasma corticosterone, and lipid peroxidation levels in the liver and heart are associated with Cal, but (ii) no additional effects were observed on the parameters that were measured when two stress regimens were given simultaneously, suggesting that animals subjected to two stress regimens can protect themselves by controlling their stress-related thresholds of response through adaptation.

Pregnenolone protects mouse hippocampal (HT-22) cells against glutamate and amyloid beta protein toxicity.

In the present work we have examined whether the neurosteroid pregnenolone has any neuroprotective effects against glutamate and amyloid beta protein neurotoxicity using immortalized clonal mouse hippocampal cell line (HT-22). The neurosteroid pregnenolone protects HT-22 cells against both 5 mM glutamate and 2 microM amyloid beta protein induced cell death in a concentration dependent manner. Optimum protection was attained at 500 nM pregnenolone, against both 5 mM glutamate as well as 2 microM amyloid beta protein induced HT-22 cell death. Furthermore, using confocal immunoflourescence microscopy we observed that 20 hours of treatment with 5 mM glutamate resulted in intense nuclear localization of the glucocorticoid receptor (GR) in HT-22 cells as compared to control untreated cells. Interestingly, 500 nM pregnenolone treatment for 24 hours, followed by 20 hours treatment with 5 mM glutamate resulted in dramatic reduction in GR nuclear localization. These results show that (i) pregnenolone has neuroprotective effects against both glutamate and amyloid beta protein neuropathology and (ii) prevention of glucocorticoid receptor (GR) localization to the nucleus may be involved in the observed neuroprotective effects of pregnenolone against glutamate neurotoxicity.

The environmental estrogenic compound bisphenol A exerts estrogenic effects on mouse hippocampal (HT-22) cells: neuroprotection against glutamate and amyloid beta protein toxicity.

We have examined using immortalized clonal mouse hippocampal cell line (HT-22) whether the environmental estrogenic compound bisphenol A (BPA), like estrogen, has any neuroprotective effect against glutamate and amyloid beta protein-induced neurotoxicity. BPA protects HT-cells against both 5 mM glutamate and 2 microM amyloid beta protein-induced cell death in a dose dependent manner. Optimum protection was attained at 1 microM and 500 nM BPA against 5 mM glutamate and 2 microM amyloid beta protein-induced HT-22 cell death, respectively. Using confocal immunoflourescence microscopy technique, we observed that 20 h of treatment with 5 mM glutamate resulted in intense nuclear localization of the glucocorticoid receptors (GR) in HT-22 cells as compared to control untreated cells. Interestingly, 1 microM BPA treatment for 24 h, followed by 20-h treatment with 5 mM glutamate, resulted in dramatic reduction in GR nuclear localization. We conclude that: (i) BPA mimics estrogen and exerts neuroprotective effects against both neurotoxins used; (ii) BPA inhibits enhanced nuclear localization of GR induced by glutamate; and (iii) HT-22 cells provide a good in vitro model system for screening the potencies of various environmental compounds for their estrogenic activity.

Biological effects of single and repeated swimming stress in male rats: beneficial effects of glucocorticoids.

We have examined the biological effects of single (45 min at 22 degrees C) and repeated swimming stress (45 min at 22 degrees C for 7 d) using male Sprague-Dawley rats. Repeated swimming for a week resulted in a significant inhibition in total body weight (25%) as compared to control unstressed animals. There was significant increase in adrenal and kidney relative weight and decreases in relative thymus weight in repeated swimming-stressed animals as compared to control animals. Repeated swimming stress resulted in almost threefold increase in plasma corticosterone levels with concomitant dramatic decrease in total glucocorticoid receptor (GR) levels in liver, thymus, and heart as compared to control unstressed animals. Interestingly, single swimming stress resulted in a significant elevation in lipid peroxidation levels in the liver and heart. In contrast, there was no change in the lipid per oxidation levels in the liver and heart between chronic stressed and control unstressed animals. Finally, both single and repeated swimming-stress animals had almost 50% reduction in plasma triglyceride levels as compared to control unstressed animals. It is concluded that elevated plasma corticosterone levels by downregulating GR during repeated swimming stress exerts beneficial effects in rats by retarding the total body weight gain and lowering plasma triglyceride levels without affecting free-radicals-induced oxidative stress.

Various types of glomus anastomoses in developing rat fetuses.

Some observations on arteriovenous anastomosis (AVA) morphogenesis of rats were considered in this investigation. Interesting typical glomus anastomoses observed under light microscopy were determined by photography before evaluation. AVAs in the skin and mesentery were compared with those reported in morphological studies in the literature. Most of the glomus anastomoses were determined to be in conformity with those in the studies reported. Some resembled simple shunts and others complex glomus anastomoses. It is suggested that the existence of various types of anastomoses of rat fetuses during development is dependent on blood flow.

Anti-stress effects of dehydroepiandrosterone: protection of rats against repeated immobilization stress-induced weight loss, glucocorticoid receptor production, and lipid peroxidation.

In the present study, we have (i) examined the biological effects of repeated immobilization stress, and (ii) tested the hypothesis that the adrenal steroid hormone dehydroepiandrosterone (DHEA) is an anti-stress hormone, using male Sprague-Dawley rats. Rats (N = 6) were divided into the following four groups: (i) control, (ii) repeated immobilization stress (2 hr daily, for 60 days), (iii) repeated immobilization stress (2 hr daily, for 60 days) plus daily i.p. administration of 5 mg DHEA/0.1 mL DMSO, and (i.v.) daily i.p. administration of 5 mg DHEA/0.1 mL DMSO alone. Results obtained showed that repeated immobilization stress resulted in a significant (25%) inhibition in body weight gain, a significant increase in adrenal weight, an increase in glucocorticoid receptor (GR) in the liver, thymus, and spleen, decreased plasma triglyceride levels, and increased lipid peroxidation in the liver and heart as compared with control unstressed animals. Interestingly, DHEA administration resulted in a significant reversal in stress-induced inhibition in body weight gain, adrenal weight, GR levels in liver, thymus, and spleen, and lipid peroxidation levels in the liver and heart. In addition, animals treated with DHEA alone without stress showed a significant (15%) inhibition in body weight gain and an almost 60% decrease in plasma triglyceride levels as compared with control unstressed animals. It is concluded that DHEA acts as an anti-stress hormone in rats, as shown in its antagonizing the effects of repeated immobilization stress on total body weight, adrenal weight, GR levels, and free radical generation.

Blastocyst implantation in rats: a morphological approach.

Very complex cellular events take place at the morphological and molecular levels both in the maternal and embryonic sites during blastocyst implantation. Although the morphological alterations in the gestational stages of the implantation site are well-known, mechanisms of the interaction between blastocyst and endometrial epithelium (apposition and adhesion stages) and penetration of the blastocyst into the endometrium have not yet been fully described. Therefore the present study was designed to determine these events morphologically throughout the implantational stages. To do this the endometrial tissues containing implantation sites from pregnant rats were taken 5 and 8 days after fertilization. In addition, non-pregnant rat endometrial tissues were also obtained in order to use as control. Tissues were processed for light and electron microscopy. Semi-thin and ultra-thin sections were stained using appropriate staining methods, and were evaluated under light and electron microscope. In the present study, interaction between trophoblast and luminal epithelium, penetration of the blastocyst into the endometrium, differentiation of germinal layers in the blastocyst and tissue alterations such as decidualization in the maternal site have been demonstrated morphologically. In conclusion, while blastocyst was implanted into the endometrium, distinct morphological changes occurred both in maternal and embryonic sites. The following stages of implantation were the development of germinal layers in the embryo and morphological changes in the maternal site leading to the placentation.

Development of arteriovenous anastomoses in Rattus norvegicus species during the fetal period.

In this study carried out on the development of arteriovenous anastomoses (AVAs) in Rattus norvegicus species, the fetus was followed up 10 days after post coitum (pc). The sections obtained from the fetuses of 10-20-day rats were subjected to the usual processes for examination under the light microscope. Various staining processes were applied on the preparations and the simple and glomus types of AVAs determined in the mesenchyme of fetuses observed to birth every day. The regions with AVAs were then photographed by photomicroscopy and evaluated. From the 10-day embryo to birth, an increase in the smooth muscles in the wall structure of simple anastomoses was observed. It was however very difficult to follow up this increase in glomus anastomoses because the wall structure is more complicated. Their morphology was examined and compared with that in literature.

[Proteus syndrome. Clinical and differential diagnostic markers]. / Proteus-Syndrome. Klinische und differentialdiagnostische Merkmale.

A case with various skeletal abnormalities, soft tissue masses and skin lesions diagnosed as Proteus syndrome is reported. Proteus syndrome is included in the differential diagnosis of many disorders, because of the polymorphous nature of its clinical manifestations. It is a rare disorder and leads to functional disabilities as well as cosmetic and psychological problems. Early diagnosis and careful follow-up with appropriate surgical interventions is important in preventing long-term complications.

[Lipoid proteinosis in 2 sisters]. / Lipoidproteinose bei zwei Schwestern.

Lipoid proteinosis was diagnosed in two daughters of a consanguinous marriage on the basis of genetic, clinical, light microscopic and ultrastructural findings. Hyaline material accumulation, thickening of the basal laminae and the resulting typical onion skin phenomenon were observed. In addition to the pathognomonic cutaneous mucosal findings, unusual manifestations such as persistence of deciduous teeth (in one case), oligodontia and intracerebral calcifications were observed. In one patient, the intracerebral calcifications caused epileptic seizures.

Ultrastructural findings of the nasal mucosa of "ozaena" in atrophic rhinitis.

Alterations in the nasal mucous layer of atrophic rhinitis "Ozaena" patients have been investigated. The vast majority (99%) of these patients were women. Morphological findings in healthy nasal mucosa demonstrated the different functional stages of the glandular tissue cells with the healthy epithelium. On the other hand, nasal mucosal material taken from the patients displayed prominent epithelial deterioration, decrease in and loss of cilia, increase in goblet cells with squamous epithelial metaplasia. A decrease in secretory granules and membrane deterioration in the apical region of the secretory cells was also clearly visible. It is possible to postulate that the ultrastructural changes seen in the secretory and storage cycles of glandular tissue of the nasal mucosa from patients seem to arise as a reaction to the superficial epithelial deterioration of the nasal mucosa together with the resulting deterioration of physiological conditions.

Renal amyloidosis: immunofluorescence and electron microscopy studies.

Renal biopsy specimens of 15 patients with renal amyloidosis were studied by immunofluorescence microscopy. The amyloidosis was associated with chronic pulmonary disease in five, rheumatoid arthritis in one, chronic lymphocytic leukemia in one, and familial Mediterranean fever in five patients. In three patients no associated condition could be determined although the pattern of organ involvement resembled that of secondary amyloidosis. IgG and complement (C3) were demonstrated in the glomerular capillary walls and in the mesangium in all patients. The pattern of the deposits was neither granular nor linear. Ig and C3 appeared as large confluent masses or broad ribbon-like segments. In the six patients studied by electron microscopy the fibrillary formation of amyloid was seen in the mesangium and the glomerular capillary walls corresponding to the Ig deposits. No immunofluorescence or ultrastructural differences were observed among the patients with secondary, inherited and leukemia-associated amyloidosis included in this study.