The epigenetic effect of nicotine on dopamine D1 receptor expression in rat prefrontal cortex

Nicotine is a highly addictive drug and exerts its effect partially through causing dopamine release, thereby increasing intrasynaptic dopamine levels in the brain reward systems. Dopaine D1 receptor (DRD1) mRNAs and receptors are localized in reward‐related brain regions, which receive cholinergic input. The aim of this study is to evaluate whether nicotine administration affects the expression of DRD1s, and if so, whether epigenetic mechanisms, such as histone acetylation, are involved. Twenty Male Sprague Dawley rats received nicotine (0.4 mg/kg/day, s.c.) or saline injections for 15 days. After nicotine/saline treatment, rats were perfused with saline; prefrontal cortex (PFC), corpus striatum (STR), and ventral tegmental area (VTA) were dissected. Homogenates were divided into two parts for total RNA isolation and histone H4 acetylation studies. DRD1 mRNA expression was significantly higher in the PFC of the nicotine‐treated group compared with controls; similar trends were observed in the VTA and STR. To study epigenetic regulation, the 2kb upstream region of the DRD1 gene promoter was investigated for histone H4 acetylation in PFC samples. After chromatin immunoprecipitation with anti‐acetyl histone H4 antibody, we found an increase in histone acetylation by two different primer pairs which amplified the ?1365 to ?1202 (P < 0.005) and ?170 to +12 (P < 0.05) upstream regions of the DRD1 promoter. Our results suggest that intermittent subcutaneous nicotine administration increases the expression of DRD1 mRNA in the PFC of rats, and this increase may be due to changes in histone H4 acetylation of the 2kb promoter of the DRD1 gene. Synapse 67:545–552, 2013. © 2013 Wiley Periodicals, Inc.     

Polymorphisms in the tumor necrosis factor-alpha gene in Turkish women with pre-eclampsia and eclampsia

The genetic background predisposing pregnant women to pre-eclampsia/eclampsia (PE/E) is still unknown. The aim of the current study was to investigate whether there is an association between the TNF-alpha-308 and 850 polymorphisms and PE or eclampsia. In this study, 40 cases of eclampsia, 113 cases of PE and 80 normotensive control cases were genotyped for the TNF-alpha-G-308A and C-850 polymorphisms. At position 308, the replacement of Guanine with Adenosine was denoted as TNF2. We found a significant difference between the TNF2 allele frequencies of the eclamptic, pre-eclamptic and normotensive controls. TNF2 (AA) polymorphism frequency was significantly higher among the eclamptics and pre-eclamptics (control : 5%, PE : 13.3%, E : 12.9%). A significantly different genotype distribution of C-850T polymorphism was observed between the PE/E and control groups, with the frequency of the variant TT genotype being significantly reduced in the preeclamptics (PE : 17% ; E : 17.5%) when compared with the control group (24.3%). We have demonstrated an association between TNF-alpha polymorphisms and pre-eclampsia susceptibility. However, it is not known whether C-850T polymorphism has a functional effect on the TNF-alpha gene. In addition, it was not possible to determine whether this polymorphism promotes the progression from PE to eclampsia because of no statistically significant difference between eclampsia and the controls.     

Comparison of different methods to detect <Emphasis Type="Italic">Helicobacter pylori</Emphasis> in the dental plaque of dyspeptic patients

The aim of this study was to compare different methods of detection of Helicobacter pylori (H. pylori) in the dental plaque of dyspeptic patients. After recording the clinical indices, culture and polymerase chain reaction (PCR) methods were performed on plaque samples, while rapid urease test in addition to these tests was carried on gastric samples from 67 dyspeptic patients who attended for an upper gastrointestinal endoscopy. Forty-seven of 67 patients were H. pylori-positive in gastric biopsy material whereas the microbial dental plaque from 19 patients demonstrated H. pylori positivity detected by PCR. Among the patients, 25.4% harbored H. pylori both in the stomach and in microbial dental plaque. No significant correlations were found among the presence of H. pylori in the stomach, in plaque, and clinical variables (P > 0.05). Although oral hygiene was observed optimal and the mean of pocket depth was not found to be higher, the prevalence of H. pylori was observed to be higher in dental plaque. According to our results, PCR technique gave the highest detection rate both in gastric biopsy and in dental plaque compared to the other methods used.     

Comparison of the proliferative activity in gingival epithelium after surgical treatments of intrabony defects with bioactive glass and bioabsorbable membrane

Guided tissue regeneration is based on preventing the more rapidly proliferating epithelium from growing into the periodontal defect after surgical procedures incorporating barrier membranes. The aim of this study was to compare the proliferative activity of gingival epithelium using proliferating cell nuclear antigen (PCNA) as a marker of cell proliferation after surgical treatments with bioactive glass graft material and bioabsorbable membrane. Using split mouth design, 20 intrabony defects were randomly assigned treatments with bioactive glass (BG group) or bioabsorbable membrane (BM group). Gingival biopsies were taken at preoperative and postoperative 12 weeks. After histological processing, the number of the inflammatory cells was measured in hematoxylin and eosin-stained sections; PCNA expression was determined in immunohistochemically-stained sections. At postoperative 12 weeks, the number of the inflammatory cells was significantly decreased (p < 0.01), PCNA expression was significantly increased (p < 0.001) in both treatment groups compared to baseline data. There was no significant difference in PCNA expression between baseline values of two groups (p > 0.05), while at postoperative 12 weeks, increase in BG group was significantly greater than that in BM group (p < 0.001). These results suggest that epithelial cell proliferation is more prominent after treatment of intrabony defects with bioactive glass compared to the treatment with bioabsorbable membrane.     

Ocular injuries,attention deficit and hyperactivity disorder,and maternal anxiety/depression levels:Is there a link?

Purpose:Given the increased risk of accidents in patients with attention deficit and hyperactivity dis-order(ADHD)or maternal anxiety/depression,we aimed to investigate the frequency of the two diseases in children with penetrating eye injury(PEI).Methods:Altogether 79 children,39 with PEIs and 40 healthy individuals(control group),aged 5-15 years,underwent a complete ophthalmologic examination.Afterwards,schedule for affective disorders and schizophrenia for school-aged children was conducted to assess the psychiatric diagnosis of all children.Turgay diagnostic and statistical manual of mental disorders(DSM-Ⅳ)-based child and adolescent behavior disorders screening and rating scale(T-DSM-Ⅳ-S)was flled by parents to evaluate the severity of ADHD symptoms.The depression and anxiety levels of mothers of each group were evaluated by two self-report measures:the Beck depression scale and the state-trait anxiety inventory(STAI),respectively.Data were analyzed by IBM SPSS version 22.0.The Chi-square and Fisher's exact test were used to determine whether there is a significant difference between qualitative variables while independent sample t and Mann-Whitney U tests to compare quantitative variables.Results:The only diagnostic difference was a significantly higher frequency of ADHD among patients with PEIs(48.7%in PEI vs.17.5%in control group,χ^2=7.359,p=0.007).The total scores of the T-DSM-Ⅳ-S(attention subscale U=418.000,p=0.006;hyperactivity subscale U=472.000,p=0.022)and maternal state-trait anxiety inventory(maternal STAI-state U=243.00,p=0.003;maternal STAl-trait U=298.000,p=0.021)were significantly higher in the PEI group than in control group.In logistic regression,children with PEI had a tendency to have a 3.5-fold increased risk for ADHD(OR=3.538,CI=0.960-13.039,p=0.058).Conclusion:ADHD was detected almost 1 in 2 children with PEls.Besides,the maternal anxiety level was significantly higher in the PEI group than in the control group.This association should be further explored via a future prospective longitudinal study.Since a proper treatment of ADHD in children and anxiety treatment in mothers may prevent vision loss following PEIs in children.     

Surgical rehabilitation following ocular chemical injury

Purpose: The purpose of this study is to evaluate the management of limbal stem cell deficiency (LSCD) secondary to chemical ocular burns.

Materials and methods: The charts of 48 eyes of 40 patients with grade 2 or higher chemical injury were evaluated retrospectively. Subjects with follow-up longer than 1 year were included. Medical treatment, surgical correction of abnormalities of ocular adnexial structures, limbal stem cell transplantation from patient’s fellow eye, from living relatives or from cadaveric donor, amniotic membrane transplantation, conjunctival epitheliectomy, chelation with ethylenediaminetetraacetic acid and penetrating keratoplasty were the treatment modalities. Outcome measures were ocular surface stability and corrected distance visual acuity (CDVA). Failure was defined as the appearance of persistent epithelial defect (nonhealing epithelial defect for more than 2 weeks) with progressive corneal conjunctivalization/vascularization and thinning, and also progression of conjunctivalization to the central 6?mm of the cornea in eyes with subsequent keratoplasty.

Results: The mean age of 31 male and 9 female patients were 32.32?±?12.6 years. LSCD was bilateral in 8 cases. The mean follow-up was 77.2?±?35.1 months. The presentations were in acute phase in 37.5%, in subacute phase in 32.5% and in chronic phase in 30% of the patients. Only 13 of 48 (27.1%) eyes obtained sufficient ocular surface stability through medical treatment; however, only 5 of these eyes achieved CDVA of less than 0.7 logMAR. Limbal stem cell transplantation was performed in 26 eyes as conjunctival limbal autograft, living-related conjunctival limbal allograft and keratolimbal allograft or as a combination of these transplantations. At the last visit, 30 eyes (62.5%) had an intact and stable ocular surface. Clear cornea was achieved in 11 (78.6%) of 14 eyes with grade 2 injury, in 9 (60%) of 15 eyes with grade 3 injury, in 5 (50%) of 10 eyes with grade 4 injury, in 1 (16.6%) of 6 eyes with grade 5 injury and in 1 (33.3%) of 3 eyes with grade 6 injury. The CDVA that was 1.66?±?0.99 logMAR initially improved to 0.87?±?0.85 logMAR at the last visit (p?Conclusion: While patients with low-grade chemical injury seem to benefit quite well from the medical treatment, amniotic membrane transplantation, limbal graft transplantation and subsequent keratoplasty; patients with severe injuries seem to be more prone to failure after all of the available treatment modalities.     

A Study of the pupal developments of Sarcophaga argyrostoma (Robineau-Desvoidy, 1830)

Forensic Science, Medicine and Pathology - Forensic entomology is a discipline in which insect evidence is used to determine the minimum postmortem interval (PMImin). The insect evidence may also...     

Human immunodeficiency virus type-1 Tat protein induces secretory leukocyte protease inhibitor expression in African green monkey but not human cells

African monkeys are resistant to HIV-1 infection due to intrinsic restriction mechanisms found in their cells. However, although they can be infected by monkey-adapted modified HIV-1 particles that are designed to overcome known restriction factors, virus numbers drop to undetectable levels in immunocompetent animals. These results indicate the possibility of the presence of yet unidentified factor(s) that restrict HIV-1 in old-world monkey (OWM) cells after integration of the viral genome into the host cell chromosome. In the light of these findings, we hypothesized that OWMs might have evolved resistance mechanism(s) against HIV-1 by switching specific gene(s) on in response to the synthesis of viral proteins in infected cells. In an attempt to mimic post-infection status, we expressed HIV-1 Tat gene in African green monkey cells and compared the whole proteome with normal cells and identified secretory leukocyte protease inhibitor (SLPI), a protein with known extracellular anti-HIV-1 activity, as an over-expressed protein in the presence of HIV-1 Tat protein by 2D-PAGE and mass spectrometry analysis. We also showed that overexpression of SLPI in the presence of HIV-1 Tat was specific to monkey cells. Our results also suggest that SLPI had a previously undiscovered intracellular anti-HIV activity in addition to its extracellular activity.

     

Outcomes of Pediatric Fistulising Perianal Crohn’s Disease

Background: Perianal disease is reported more widely in pediatric Crohn patients than in the past, and has been stated as an independent modifier of the disease behavior. In this study, we aimed to analyze the clinical characteristics and outcomes of fistulising perianal Crohn’s disease (fpCD) in the pediatric age group.Methods: A total number of 149 children with an established diagnosis of inflammatory bowel disease who have been diagnosed before 18 years of age and followed in our tertiary center were revised. Clinical, endoscopic, laboratory, and radiologic data of 50 patients with CD, who had at least 18 months follow-up data, were compiled.Results: Of 50 patients, 26 (52%) were diagnosed as fpCD (38% at onset). More than half of the patients without any notable external orifices around the perianal area were diagnosed as fpCD by an magnetic resonance imaging (MRI). Pediatric fpCD patients had a higher disease activity score and platelet count, lower serum albumin level, and a higher rate of granuloma in the biopsy samples, compared with non-fistulising patients. A considerably high rate of surgical interventions (i.e., seton placement 46% and abscess drainage 15%) was performed in combination with infliximab.Conclusion: Fistulising perianal Crohn’s disease seems to be more common than previously reported in the pediatric age group. A severe course of the disease might serve as a warning for the development of fpCD. A careful physical examination and use of perianal MRI with a high index of suspicion may increase the likelihood of fistula detection, hence may change the treatment strategy.     

The effect of smoking on epithelial proliferation in healthy and periodontally diseased marginal gingival epithelium

BACKGROUND: Smoking causes an increase in the thickness of gingival epithelium, which is the outcome of increased keratinocyte proliferation or loss. Smoking-related changes in the proliferative activity of the gingival epithelium are largely uncharacterized for periodontal diseases. The aim of the present study was to determine the effects of smoking on the proliferation of the epithelium in periodontally diseased marginal gingiva by comparing the expression patterns of two different proliferation markers. METHODS: Gingival biopsies (N=60) were obtained from smokers who had clinically healthy gingiva (n=10), smokers with gingivitis (n=10), smokers with periodontitis (n=10), non-smokers with clinically healthy gingiva (n=10), non-smokers with gingivitis (n=10), and non-smokers with periodontitis (n=10). The quantitative measurement of maximum epithelial thickness was performed on hematoxylin and eosin-stained sections. The expression patterns for proliferating cell nuclear antigen (PCNA) and Ki67 were evaluated immunohistochemically. RESULTS: The percentage of PCNA-positive cells was higher than the percentage of Ki67-positive cells in all groups (P<0.001). When the mean values of PCNA and Ki67 were compared in each group, a statistically significant difference was observed only in the healthy smoker group (P=0.003). Significant differences in PCNA proliferation indices were only found between the smoker group and the non-smoker healthy group (P=0.015). CONCLUSIONS: Smoking had an affect on the proliferation of cells in the oral gingival epithelium, regardless of periodontal status. The increase in thickness of the epithelium was not associated with smoking; periodontal status and inflammation seemed to be more important factors. Smoking induced the replication activity of gingival epithelium and induced DNA repair.