The influence of post-fixation on visualising vimentin in the retina using immunofluorescence method.

BACKGROUND: Post-fixation of sections is especially required for cryostat sections of fresh frozen tissues. Vimentin is an intermediate filament in both fibrillary and non-fibrillary form, expressed in Müller's cells and astrocytes of the retina. Our aim was to determine the best post-fixation method for visualising vimentin in archival mouse eyes. MATERIALS AND METHODS: We used an archival mouse eye, slightly pre-fixed with paraformaldehyde and stored at -80°C for 4 years. We tried three fixatives (pa-raformaldehyde [PFA], alcohol/acetic acid [AAA] and methanol) for post-fixation of eye sections. RESULTS: We showed that post-fixation alters the labelling properties of vimentin expressed in the retina. In the sections with no post-fixation, vimentin positivity was observed in and around the nuclei in non-fibrillary form. In PFA post-fixed sections, the vimentin in the retina was not observed as fibrils. Positivity was observed in the nuclei and in perinuclear regions of the cells. In AAA post-fixed sections, positive labelling was observed around the nuclei as fibrils. In methanol post-fixed sections, labelling was observed around the nuclei as fibrils. CONCLUSIONS: We conclude that post-fixation with AAA is more convenient for immunofluorescent labelling of vimentin in the retina for slightly PFA pre-fixed and long-term stored retina. (Folia Morphol 2018; 77, 2: 246-252).

The Effects of Gonadotropin Replacement Therapy on Metabolic Parameters and Body Composition in Men with Idiopathic Hypogonadotropic Hypogonadism.

Testosterone replacement therapy (TRT) in idiopathic hypogonadotrophic hypogonadism (IHH) slows the process of metabolic syndrome (MetS), diabetes mellitus, and cardiovascular diseases by its inversing effects on insulin resistance, dyslipidemia, and blood pressure. Since there are not enough data regarding the effects of gonadotropin replacement therapy (GRT), we aimed to investigate the impact of GRT on MetS parameters in IHH patients. Sixteen patients with IHH and 20 age and body mass index (BDI)-matched healthy controls were enrolled into the study. Patients were evaluated at baseline and 6 months after the GRT. Sex hormones, insulin like growth factor-1, prolactin, insulin, C-reactive protein (CRP), homocysteine, and lipid levels were measured at baseline and after the treatment. Anthropometric measurements, including BMI, body fat ratio (BFR), fat free mass (FFM), waist circumference, and waist-to-hip ratio (WHR), were also performed. Homeostasis Model Assessment of Insulin Resistance (HOMA-IR) index was calculated. Body fat ratio, triglyceride, HOMA-IR, and CRP levels were higher, whereas bone age, fat free mass, and creatinine levels were lower in the patients with hypogonadism. HOMA-IR indices and basal insulin levels decreased significantly after 6 months of GRT compared with baseline levels. Triglyceride levels, and BFRs diminished significantly by an accompanying decline in WHR. FFM of the patients increased following the GRT. No significant changes were detected in CRP, homocysteine, total and LDL-cholesterol levels. Similar to TRT, hCG treatment decreases HOMA-IR, triglyceride levels, BFR and WHRs, and increases FFM in patients with IHH.

The impact of the time interval on in-vitro fertilisation success after failure of the first attempt.

The aim of this study was to identify the optimal time interval for in-vitro fertilisation that would increase treatment success after failure of the first attempt. This retrospective study evaluated 454 consecutive cycles of 227 infertile women who had two consecutive attempts within a 6-month period at an IVF centre. Data were collected on duration of stimulation, consumption of gonadotropin, numbers of retrieved oocytes, mature oocytes, fertilised eggs, good quality embryos on day 3/5 following oocyte retrieval and clinical and ongoing pregnancy. There were significant increases in clinical pregnancy rates at 2-, 3- and 4-month intervals. The maximum increase was after two menstrual cycles (p = 0.001). The highest rate of ongoing pregnancy was in women that had the second attempt after the next menstrual cycle following failure of IVF (27.2%). After IVF failure, initiating the next attempt within 2-4 months increases the clinical pregnancy rates.

Therapeutic approach to patients with familial Mediterranean fever-related amyloidosis resistant to colchicine.

The frequency of FMF-related amyloidosis has been decreased by colchicine use over the past few decades. However, the beneficial effect of colchicine may differ in accordance with nephropathic stages. When used in proper doses and with compliance, colchicine is very effective in preclinical and proteinuric stages of FMF-related amyloidosis. Even so, a large number of patients with nephrotic range proteinuria, despite compliance and an ideal dose of colchicine, may still progress to end-stage renal failure (ESRF).We do not know exactly what we can do with such patients. This paper discusses the therapeutic approach to patients with FMF-related amyloidosis.

Myocardial infarction in young patients (< or =35 years of age) with systemic lupus erythematosus: a case report and clinical analysis of the literature.

The present study aims to report a-20-year old girl with systemic lupus erythematosus (SLE) who developed myocardial infarction (MI) and also aims to review acute myocardial infarction (AMI) in young SLE cases (< or =35 years) reported in the literature. We conducted a comprehensive review of the English literature from 1975 to 2006 to analyse data on MI in SLE patients who had developed AMI either at 35 or earlier. In 32 English articles, we identified 49 SLE patients, plus our case, with AMI. They consist of 41 female and nine male patients, their mean age being 24 +/- 6.4 years (range of 5-35). Disease duration varied between 0 and 13 years. The lag time between the onset of the SLE manifestations and development of AMI was 7.7 +/- 5.4 year (range of 1 month to 20.5 years). We divided the patients into three subgroups according to their coronary involvement type (Group I: normal coronary artery or coronary thrombosis (n = 16); Group II: coronary aneurysm/arteritis (n = 12); Group III: coronary atherosclerosis (n = 22)). The lag time between the onset of the SLE manifestations and development of MI in the subgroups showed variations: Group I < Group II < Group III. Both prevalence of renal involvement and steroid therapy were higher in patients with coronary atherosclerosis than were in Group I. There were one or more risk factors for atherosclerosis in 39 SLE patients. AMI in young SLE patients may be seen, albeit rare. We suggest that clinicians should have a low threshold for cardiac evaluation in patients with SLE. Also, traditional risk factors could be managed through preventive measures.

Increased frequency of ultrasonographic findings suggestive of renal stones in patients with ankylosing spondylitis.

OBJECTIVE: To determine the frequency of hypercalciuria and renal stones in ankylosing spondylitis (AS) sufferers. METHODS: This study involved 83 consecutive AS patients (21 female, 62 male; mean age 36.7 yr), 72 consecutive Behcet's disease (BD) patients (29 female, 43 male; mean age 37.7 yr) as disease control and 92 healthy control (HC) (26 female, 66 male; mean age 32.9 yr.) Twenty-four hour urine analyses for urinary calcium and uric acid levels were performed in each patient. Likewise, blood samples for erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), parathyroid hormone (PTH), calcium and uric acid evaluation were taken. Renal ultrasonography to evaluate the presence of renal stone was performed in patients with AS, as well as in the BD patients and HC individuals after a fasting period of 12 hours. RESULTS: 20 of the 80 (25%) patients with AS were diagnosed with renal stones. Only 4/72 (5.5%) BD patients, and 3/98 (3.3%) HC individuals had renal stones by ultrasonography. AS patients had a significantly higher frequency of renal stones compared with BD (p < 0.001) and HC (p < 0.0001). AS patients with renal stones were much older and their disease duration was much longer in comparison with AS patients without renal stones. Ultrasonographic and laboratory findings did not correlate. The number of AS patients with hypercalciuria who had renal stones was higher than that of AS patients who did not have renal stones (p < 0.01). There was a negative correlation between acute phase response and spinal mobility. CONCLUSION: Renal stone prevalence was found to increase in AS patients. The likelihood of renal stone formation was also found to increase with the extension of the disease duration of AS sufferers.

Potentiation of androgen receptor transcriptional activity by inhibition of histone deacetylation--rescue of transcriptionally compromised mutants.

Androgens are critical in the development and maintenance of the male reproductive system and important in the progression of prostate cancer. The effects of androgens are mediated by the androgen receptor (AR), which is a ligand-modulated transcription factor that belongs to the nuclear receptor superfamily. We and others have previously shown that CREB-binding protein (CBP) can function as a coactivator for AR. Similar to some other nuclear receptor coactivators and/or the proteins that they interact with, CBP has histone acetyl transferase (HAT) activity that is thought to contribute to transcriptional activation by nuclear receptors. We have therefore assessed whether an increase in the histone acetylation status in the cell can influence AR transcriptional activity, by using the histone deacetylase (HDAC) inhibitors (HDACIs) trichostatin A (TSA), sodium butyrate (Na-But) and depsipeptide (FR901228). We found that inhibition of HDAC activity significantly increased the ability of endogenous AR in LNCaP cells, or ectopically expressed AR in HeLa cells, to activate transcription from AR-dependent reporter constructs. In addition, HDACIs increased the androgen-dependent activation of the prostate-specific antigen (PSA) gene in LNCaP cells, an increase that was not due to an increase in nuclear AR protein levels. Moreover, the viral oncoprotein E1A that inhibits CBP HAT activity fully repressed the ability of HDACIs to stimulate AR-mediated transcription, indicating that CBP is involved in this process. Deletional mutagenesis of AR indicated that whereas the AF-2 domain in the C-terminus is dispensable, the AF-1 domain in the N-terminus is required for augmentation of AR action by HDACIs, an observation which is in concordance with the reduced ability of CBP to activate AR N-terminal deletion mutants. Furthermore, HDACI treatment rescued the deficiency in the transactivation potential of AF-2 mutants. Taken together, our findings suggest that a change in the level of histone acetylation of target genes is an important determinant of AR action, possibly mediated by CBP.

Addison's disease associated with widespread abdomino-pelvic visceral calcification due to secondary amyloidosis: a case report.

We report the case of a 42-year-old woman who presented with Addison's disease with widespread abdomino-pelvic visceral calcifications due to secondary amyloidosis. AA amyloidosis and calcification were supported by histological evidence of liver tissue. To our knowledge, no other case with such extensive visceral calcification involving the adrenals, liver, ovaries, and renal secondary to amyloidosis has been published.

Effect of mannitol in experimental spinal cord injury: an ultrastructural and electrophysiological study.

BACKGROUND: Although many experimental and clinical studies were performed on the pathophysiology and treatment of spinal cord injury (SCI), the electrophysiological and ultrastructural changes of the spinal cord were not precisely evaluated. AIMS: To investigate the effect of mannitol on Somatosensory Evoked Potentials (SSEP), postoperative neurological recovery and ultrastructural findings after an experimental SCI. SETTING: The experimental microsurgery laboratory of a university hospital. DESIGN: A prospective, randomized animal study. MATERIAL AND METHODS: Sprague-Dawley rats were used and divided into three groups (Groups I-III) for this study. Those in Group I were control animals who underwent laminectomy only, and non-traumatized spinal cord samples were obtained 2 weeks later. SCI was produced in Groups II and III using clip compression technique, and cord samples were obtained 2 weeks later. The rats in Group II received 2 g/kg of 20% mannitol intraperitoneally, immediately and three hours after trauma was induced; and those in Group III received the same amount of 0,9% NaCl in the same manner. Preoperative and postoperative SSEP records at the end of 2 weeks were obtained. Electron microscopy examination of the cord samples was done at 2 weeks postoperatively. STATISTICAL ANALYSIS USED: Fischer's Exact Test. RESULTS: SSEP records, ultrastructural findings and clinical recovery showed that minor neural damage and significant recovery occurred in Group II. CONCLUSION: This study demonstrates that the administration of 2 g/kg of 20% mannitol produces significant improvement in the neural structures and protects the spinal cord following injury.

Renal involvement in primary antiphospholipid syndrome and its response to immunosuppressive therapy.

Antiphospholipid syndrome (APS) is the association between antiphospholipid antibodies, venous and arterial thrombosis and pregnancy morbidity. Although the kidney may be affected in APS, the treatment of renal involvement is yet to be elucidated. This report describes the clinical and laboratory features of four patients with primary APS nephropathy, and the beneficial effect of immunosuppressive therapy accompanied by warfarin and angiotensin-converting enzyme inhibitor. We also briefly discuss the possible mechanisms of the beneficial effects of immunosuppressives on primary APS nephropathy.

Is there an association of plasma homocysteine levels with vascular involvement in patients with Behçet's syndrome?

OBJECTIVE: To assess whether or not plasma homocysteine levels play a part in vascular involvement in Behçet's syndrome (BS). METHODS: 74 consecutive BS patients fulfilling the criteria of the International Study Group for BS, 35 healthy control (HC) and 14 rheumatoid arthritis (RA) patients on methotrexate (MTX) were studied. BS patients were then classified as those with and without vascular involvement. Fasting plasma homocysteine, folate, and vitamin B12 concentrations were measured by enzyme immunoassay and chemiluminescent immunoassay methods respectively. RESULTS: Plasma homocysteine levels were found to be higher in the BS patients than in the healthy control (16.08 +/- 7.5 vs. 12.9 +/- 6.3 micromol/L, p < 0.03). The homocysteine levels in the RA group on MTX were higher compared with both the BS and HC groups (28.7 +/- 9.9; p < 0.0001). No remarkable difference pertaining to homocysteine levels was found between BS patients with or without thrombosis (p < 0.86). Hyperhomocysteinemia was also detected in 11 out of 22 (50%) of the patients with vascular involvement, which proved to be of no significant difference in comparison with those without vascular involvement (20/52, 38%; chi2 = 0.26, p > 0.05). Active BS smokers exhibited a higher concentration of homocysteine in contrast to non-smoker BS sufferers (20 +/- 8.4 vs 14.1 +/- 6.1 micromol/l; p < 0.004). Smoking was determined to have a positive correlation with vascular involvement (r = 0.26, p < 0.046), as well as with homocysteine levels (r = 0.31, p < 0.012) in BS. Upon logistic regression analysis, smoking was found to have a significant relationship with vascular involvement (odds ratio 3.12 [95% CI 2.02-4.22] p = 0.04). There was no significant difference between the study groups with respect to their B12 vitamin and folate levels. We were unable to make any correlation between homocysteine and vitamin B12 or folate in any of the groups (p > 0.05). CONCLUSIONS: No association was found between homocysteine levels and vascular involvement in our BS patients. We determined that smoking seems to pose a risk for vascular involvement in BS patients.