Retrospective review of re-positive qPCR tests for SARS-CoV: do they indicate presence of reinfection?

OBJECTIVE: In 2019, the Coronavirus Disease 2019 (COVID-19) pandemic broke out, caused by the coronavirus called Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). Reinfections can be observed with various respiratory viruses, including human coronaviruses. Moreover, they may result from weak or waning initial immune response, reinfection with another genotype/subtype, or the rapid antigenic changes in the virus. The aim of this study was to investigate the likelihood of reinfection in COVID-19 patients that had a positive qPCR test result at least 60 days after a negative test result in patients that were confirmed with COVID-19 on qPCR. MATERIALS AND METHODS: The quantitative polymerase chain reaction (qPCR) results of a total of 105,000 samples that had been obtained between April 1, 2020, and February 1, 2021, in two separate authorized laboratories were retrospectively analyzed. 22 samples from 11 patients included in the study, qPCR tests were repeated for each sample using the Rotorgene Q PCR system with Diagnovital SARS-CoV-2 (RTA Labs, Turkey) Real-Time PCR kits. Positive samples were screened for B.1.1.7 and E484K mutations using the qPCR method on the Rotorgene Q PCR system with Bio-Speedy SARS-CoV-2 Variant Plus kits (Bioeksen Technology, Turkey). RESULTS: The 105,000 individuals comprised 55,614 men and 49,386 women. In the qPCR test, 14,511 (13.82%) individuals were found to be positive for SARS-CoV-2. Of these, 11 (0.076%) patients were included in the study based on the inclusion criteria. Accordingly, the risk of reinfection was calculated as 0.076% (95% confidence interval [CI]: 0.056%-0.096%) and the incidence was 1.04 per 10,000 population (95% CI: 0.62-1.38 per 10,000). No patient was admitted to the intensive care unit or died during both episodes. Moreover, no B.1.1.7 or E484K mutation was detected in any patient. CONCLUSIONS: The high frequency of COVID-19 infection poses serious risks for the development of new variants and the currently used vaccines are likely to lose their efficacy against new variants. To reduce these risks and to be successful in the fight against the pandemic, we suggest compliance with personal protective measures as well as rapid and widespread application of vaccination not only in developed countries but also in the whole world and the modification of currently used vaccines in such a way to fight against newly emerged variants.

Protective effect of nebivolol on gentamicin-induced nephrotoxicity in rats.

OBJECTIVE: Nephrotoxicity is a major complication of gentamicin (GEN), which is widely used in the treatment of severe Gram-negative infections. As we know, treatment with nebivolol has been shown to decrease renal fibrosis and glomerular injury as well as improve endothelial dysfunction. Therefore, we evaluated the potential protective effect of nebivolol (NBV) against GEN-induced nephrotoxicity in rats. MATERIAL AND METHOD: Twenty-four rats were randomly divided into four groups: control group (Group 1); rats intraperitoneally injected with GEN (100 mg/kg/day; Group 2); rats treated with GEN plus distilled water (Group 3); and rats treated with GEN plus NBV (10 mg/kg/day; Group 4). After 15 days, the rats were sacrificed, their kidneys taken, and blood analysis performed. Tubular necrosis and interstitial fibrosis scores were determined histopathologically in a part of kidneys; nitric oxide (NO), malondialdehyde (MDA), and reduced glutathione (GSH) levels were determined in other part of kidneys. RESULTS: The GSH levels in renal tissue of only GEN-treated rats were significantly lower than those in control group, and administration of NBV to GEN-treated rats significantly increased the level of GSH. The group that was given GEN and NBV had significantly lower MDA and NO levels in kidney cortex tissue than that given GEN alone. Despite the presence of mild tubular degeneration, the rats treated with GEN+NBV showed a less severe tubular necrosis, and their glomeruli maintained a better morphology compared to GEN group. CONCLUSION: NBV exerts antioxidant, anti-inflammatory and antifibrotic effects on GEN-induced kidney damage by reducing oxidative stress in rat model (Tab. 3, Fig. 2, Ref. 68).

rs3918242 variant genotype frequency and increased TIMP-2 and MMP-9 expression are positively correlated with cancer invasion in urinary bladder cancer.

To study the role of MMP9 and TIMP2 genotypes and expression in predisposition to bladder cancer and relation with metastasis. 100 urinary bladder cancer patients and 100 healthy controls were included in the study. rs3918242 and rs8179090 genotypes were determined with PCR-RFLP. Quantitative real-time polymerase chain reaction was employed to assess the MMP-9 and TIMP-2 expression in tumors and adjacent healthy tissues. Variant genotype (TT) for rs3918242 polymorphism and rs8179090 variant genotype are not associated with bladder cancer risk. rs3918242 genotype was significantly associated with tumor invasion. In contrast with this, rs8179090 genotype has not shown a significant association with tumor invasion. Both SNPs did not show a significant association with metastatic status. MMP-9 was upregulated in tumors in comparison to cancer free tissues. Significant increase in the expression of MMP-9 was also observed in invasive tumors. TIMP-2 expression was significantly increased in tumors in comparison to cancer free tissues and in metastatic tumors in comparison to non-metastatic tumors. Tissues with rs3918242 variant genotype have shown increased MMP-9 expression.  rs3918242 promoter polymorphism of MMP-9 is significantly associated with tumor invasion, however; there is no positive correlation between TIMP-2 rs8179090 promoter polymorphism variant frequency and invasion. MMP-9 and TIMP-2 genes are upregulated in cancerous tissues when compared to normal bladder tissues.

Effects of taking tadalafil 5 mg once daily on erectile function and total testosterone levels in patients with metabolic syndrome.

We aimed to evaluate the efficacy of tadalafil 5 mg once-daily treatment on testosterone levels in patients with erectile dysfunction (ED) accompanied by the metabolic syndrome. A total of 40 men with metabolic syndrome were evaluated for ED in this study. All the patients received 5 mg tadalafil once a day for 3 months. Erectile function was assessed using the five-item version of the International Index of Erectile Function (IIEF) questionnaire. Serum testosterone, follicle-stimulating hormone and luteinising hormone levels were also evaluated, and blood samples were taken between 08.00 and 10.00 in the fasting state. All participants have three or more criteria of metabolic syndrome. At the end of 3 months, mean testosterone values and IIEF scores showed an improvement from baseline values (from 3.6 ± 0.5 to 5.2 ± 0.3, from 11.3 ± 1.9 to 19 ± 0.8 respectively). After the treatment, serum LH levels were decreased (from 5.6 ± 0.6 to 4.6 ± 0.5). There was significantly difference in terms of baseline testosterone and luteinising hormone values and IIEF scores (p < .05). Based on our findings, we recommend tadalafil 5 mg once daily in those men with erectile dysfunction especially low testosterone levels accompanied by metabolic syndrome.

Deregulation of seven CpG island-harboring miRNAs in bladder cancer: miR-155 and miR-23b as the most promising oncomiRs.

Analyses of differential miRNA expressions in tumor and normal tissues can identify specific miRNAs involved in cancer pathogenesis, which can then be used as diagnostic, therapeutic and prognostic biomarkers. In this respect, we aimed to investigate expression levels of seven CpG island-harboring miRNAs in 50 paired UBC tissues by qRT-PCR. miR-21 and miR-155 were found to be significantly upregulated, and miR-23b, miR-126, miR-129-5p, miR-143a and miR-218-5p were downregulated. ROC analysis indicated miR-155 as the most promising candidate for discrimination of tumors from healthy tissue, and miR-23b for the discrimination of early stage from late stage tumors.

Lipoprotein-associated phospholipase A2 levels are associated with erectile dysfunction in patients without known coronary artery disease.

Endothelial dysfunction and microvascular damage play a crucial role in the pathogenesis of erectile dysfunction (ED). Lp-PLA2 is a calcium-independent member of the phospholipase A2 family and hydrolyses oxidised phospholipids on low-density lipoprotein (LDL) particles that plays a pivotal role in ox-LDL-induced endothelial dysfunction. The purpose of the current study was to determine the association between Lp-PLA2 levels and ED in patients without known coronary artery disease (CAD). All patients were evaluated for ED and divided into two groups: 88 patients suffering from ED for >1 year were enrolled as an experimental group and 88 patients without ED were enrolled as a control group in this study. Diagnosis of ED was based on the International Index of Erectile Function Score-5. Levels of Lp-PLA2 were measured in serum by colorimetric assay. The relationship between Lp-PLA2 levels and ED in patients was evaluated statistically. The mean age of patients with ED group was 59.4 ± 11.32 and 55.8 ± 9.67 in the control group. Plasma Lp-PLA2 levels were significantly higher in ED than in the control group (220.3 ± 66.90 and 174.8 ± 58.83 pg ml(-1) , respectively, P < 0.001). The Lp-PLA2 levels were negatively correlated with score of ED (r = -0.482, P < 0.05). In logistic regression analysis, enhanced plasma Lp-PLA2 levels result in approximately 1.2-fold increase in ED [1.22 (1.25-2.76)]. In this study, serum Lp-PLA2 levels were found to be associated with endothelial dysfunction predictive of ED. Serum Lp-PLA2 level appears to be a specific predictor of ED, and it may be used in early prediction of ED in the male population.

Combination therapy with selective serotonin reuptake inhibitors and phosphodiesterase-5 inhibitors in the treatment of premature ejaculation.

We aimed to evaluate the effectiveness of paroxetine and tadalafil combination in the treatment of premature ejaculation (PE). A total of 150 primary (lifelong)PE patients were randomly distributed into three groups of 50 patients each. Group 1 received 20 mg paroxetine every day for 1 month, Group 2 received 20 mg tadalafil on demand 2 h before intercourse, and Group 3 received paroxetine and tadalafil on demand 2 h before intercourse. Intravaginal ejaculatory latency times (IELT) scores were evaluated at baseline, at the end of the first month of therapy and 1 month after discontinuation of the treatment, while International Index of Erectile Function (IIEF) questionnaire scores were evaluated both prior to and after the treatment. At the end of the first month of therapy, IELT scores were compared with the basal values and statistically significant changes were detected (60.6 ± 30.2-117.3 ± 67.3, 68.5 ± 21.4-110.2 ± 37.3, 71.56 ± 40.23-175.2 ± 60.2)(P < 0.01). IELT scores after discontinuation of treatment were found to be close to the baseline IELT scores (P > 0.05). IIEF scores were evaluated both prior to and after the treatment, and no statistically significant difference was detected (P > 0.05). It is concluded that utilisation of selective serotonin reuptake inhibitors (SSRI) and phosphodiesterase-5 inhibitors (PDE5i) combination before intercourse seems to provide significantly longer ejaculatory latency times as compared with SSRI alone for a long time in patients with PE.

Purification and characterization of an extremely stable glucose isomerase from Geobacillus thermodenitrificans TH2.

The D-glucose/D-xylose isomerase was purified from a thermophilic bacterium, Geobacillus thermodenitrificans TH2, by precipitating with heat shock and using Q-Sepharose ion exchange column chromatography, and then characterized. The purified enzyme had a single band having molecular weight of 49 kDa on SDS-PAGE. In the presence of D-glucose as a substrate, the optimum temperature and pH of the enzyme were found to be 80 degrees C and 7.5, respectively. The purified xylose isomerase of G. thermodenitrificans TH2 was extremely stable at pH 7.5 after 96 h incubation at 4 degrees C and 50 degrees C. When the thermal stability profile was analyzed, it was determined that the purified enzyme was extremely stable during incubation periods of 4 months and 4 days at 4 degrees C and 50 degrees C, respectively. The K(m) and V(max) values of the purified xylose isomerase from G. thermodenitrificans TH2 were calculated as 32 mM and 4.68 micromol/min per mg of protein, respectively. Additionally, it was detected that some metal ions affected the enzyme activity at different ratios. The enzyme was active and stable at high temperatures and nearly neutral pHs which are desirable for the usage in the food and ethanol industry.

Stress urinary incontinence in premenopausal and postmenopausal women: evaluation of serum estradiol levels and bone mineral density.

AIM: Stress urinary incontinence is the most common form of urinary incontinence, occurring in pure or mixed forms in nearly 80% of women with incontinence. Hypoestrogenism may cause female incontinence and low bone mineral density, together. So, we investigated the relationship between stress urinary incontinence, serum E2 levels and osteoporosis in premenopausal and postmenopausal women. METHODS: From February 2011 to March 2012, 78 postmenopausal and 30 premenopausal women with stress incontinence, and 57 continent postmenopausal and 20 premenopausal women included in the study. All women's ages, body mass indexes, comorbidities, numbers of birth, number of pregnancies, serum estradiol levels and T-scores were evaluated and compared between groups. RESULTS: Bone mineral density was evaluated in groups. Osteoporosis was more in women with stress urinary incontinence (P<0.05). E2 levels were found decrease in the postmenopausal and premenopausal women who have stress urinary incontinence compared to control group. we found that the women who have low estradiol levels, usually T score was ≤ -2.5 and have osteoporosis. CONCLUSION: Osteoporotic women should be evaluated for urinary incontinence and vice versa women with urinary incontinence evaluated for osteoporosis. Further studies are needed to clarify molecular mechanisms of these results.

Lamivudine and Adefovir Motif Variants Detected in chronic Hepatitis B patients.

BACKGROUND AND AIMS: Chronic hepatitis B is an important health problem worldwide. Lamivudine, adefovir, entecavir and telbivudine are the oral drugs licensed for the treatment of patients with chronic hepatitis B. Implementation of antiviral therapy leads to the emergence of mutant strains during the treatment in chronic hepatitis B. Primary antiviral resistance may be rarely encountered. The aims of this study were to detect the resistance patterns of Hepatit B Virus strains in treatment-naive chronic hepatitis B patients. MATERIALS AND METHODS: A total of 147 CHB patients were included to this study which was carried on between January 2007- December 2010. HBV DNA levels were detected by using the Real time PCR (COBAS Ampli- Prep/COBAS TaqMan HBV Test). HBV-DNA was extracted from the sera of the patients by using extraction kit (Invisorb, Instant Spin DNA/RNA Virus Mini Kit, Germany). A line prob assay (Inno-Lipa HBV DR v2, Innogenetics N.V, Ghent, Belgium) was used to determine motif variants at viral polymerase gene fragment in HBV-DNA samples of these patients and evaluated colorimetrically. RESULTS: In 147 patients antiviral resistance rate was found 17% (25/147) for lamivudin, 5.44% (8/147) adefovir, 0.68%(1/147) lamivudin and adefovir. Various mutations were detected. This mutations; responsible for lamivudine resistance YMDD+YVDD (n=10), YMDD+YIDD (n=12), YIDD (n=2), YVDD (=1); responsible for adefovir resistance N236T (n=3), A181T (n=5); responsible for lamivudine and adefovir resistance YMDD+YIDD+N236T (n=1). CONCLUSIONS: As a conclusion, it is thought that drug resistance should be followed up regularly, the determination of HBV drug resistance as immediate as possible period may be instructive for the treatment and follow-up in CHB patients. Although determination of known mutations with Inno Lipa DR v2 method is disadvantage, because of ease of application and the determination of both lamivudin-adefovir resistance in a short time, it can be used for the treatment and follow-up in CHB patients.

Relevance of serum nitric oxide levels and the efficacy of selective serotonin reuptake inhibitors treatment on premature ejaculation: decreased nitric oxide is associated with premature ejaculation.

The aim of the present study was to determine the relevance of serum nitric oxide levels and the efficacy of selective serotonin reuptake inhibitors (SSRI) treatment on premature ejaculation. Sixty married men (aged 20-50) with lifelong premature ejaculation and forty healthy men (aged 24-48) as control group were included in this study. The patients were evaluated by intravaginal ejaculation latency time (IELT) for premature ejaculation (PE). IELT<1 min is accepted PE. Patients with diabetes mellitus, chronic disorders or erectile dysfunction and heavy smokers were excluded. All patients were evaluated with history, physical examination, International Index of Erectile Dysfunction-5 (IIEF-5) score and IELT by stopwatch method. Nitric oxide levels were measured by Griess reaction, and all samples were frozen at -80 °C. Patients were randomly categorised 4 group to receive fluoxetine 20 mg day(-1) (Group 1), paroxetine 20 mg day(-1) (Group 2), sertraline 50 mg day(-1) (Group 3) and healthy control (Group 4) for 4 weeks. Baseline and post-treatment findings were compared between the four groups. At the end of 4 weeks, in fluoxetine, paroxetine, sertraline groups mean IELT values showed a statistically significant improvement from the baseline values (P < 0.001, P < 0.001, P = 0.03; respectively). Baseline and 1st month follow-up mean IIEF scores were 24.5 and 23.05, 24.70 and 23.60 (P < 0.05) in group 1 and group 3 respectively; also 23.09 and 23.32 (P > 0.05) in group 2. Baseline serum NO levels were 31.8, 30.44, 30.8 and 42.84 in fluoxetine, paroxetine, sertraline and healthy control groups respectively. NO levels were statistically lower in patients with PE. After treatment of fluoxetine, paroxetine and sertraline, NO levels were increased baseline (35.8, 36.4, 38.08) (P < 0.05). Our findings indicated that PE is associated with decreased serum NO levels. After the SSRI treatment increased, NO may retard ejaculation presumably by central peripheral mechanism. Further studies are needed to confirm this suggestion and the role of NO in pathophysiology and treatment for premature ejaculation.

Effect of tadalafil and 3-hydroxy-3-methylglutaryl coenzyme A inhibitor statin on the haemodynamics of cavernous and brachial arteries.

Erectile dysfunction (ED) is usually associated with cardiovascular disease and reduced endothelial function. The aim of the present study was to examine the effect of tadalafil and statin on the endothelial function of cavernous and brachial arteries in healthy men and in patients with ED. The cases included in the study were as follows: 150 men with ED complaints for at least 6 months, and 50 healthy volunteers without sexual problems. Patients were randomly divided into four groups of equal numbers. Group 1 received 20 mg of tadalafil on alternate days, Group 2 received 10 mg of statin a day, Group 3 received tadalafil on alternate days and 10 mg of statin a day, and the last group served as controls. Noninvasive evaluation of brachial artery flow-mediated dilatation (FMD) and percentage of increase in cavernosal arteries diameter (PICAD) was conducted via ultrasound at baseline and 4 weeks after administration of tadalafil or atorvastatin. Before drug administration, FMD and PICAD values did not significantly differ among the three treatment groups. After drug administration, FMD and PICAD values significantly increased in patients receiving tadalafil and tadalafil+statin (P < 0.001), but not in patients receiving only statin. These findings suggested that use of tadalafil alone and tadalafil combined with statin improved endothelial function of cavernous and brachial arteries.

Potential role of p38-mitogene-activated protein kinase and nuclear factor-kappa B expression in testicular dysfunction associated with varicocele: an experimental study.

The aim of this study was to investigate p38-mitogene-activated protein kinase (p38-MAPK), nuclear factor-kappa B (p65-NF-kB) and inducible nitric oxide synthase (iNOS) expression in an experimental model of varicocele in the rat testis. Male Wistar albino rats (n = 18) were divided into three equal groups: control group, sham operated group and left varicocele-induced group. Malondialdehyde (MDA), nitric oxide (NO) and reduced glutathione (GSH) levels were biochemically assessed, and the p38-MAPK and NF-kB activity, and iNOS expression were immunohistochemically studied in the right and left testicles of rats from each group. The GSH levels were significantly decreased, whereas the level of MDA and NO was significantly increased in the testicular tissues of rats in varicocele group compared with those of the control and sham groups. There was a marked staining for iNOS, p38-MAPK and p65-NF-kB expression in rats of varicocele group compared with the sham group. There was no positive staining in rats of control group. There were significant differences in biochemical, histological and immunohistochemical studies, but no significant differences were noted between other groups. p38-MAPK and p65-NF-kB activation, and iNOS expression have a significant role in varicocele-induced testicular dysfunction.

Castleman's disease as cervical mass: a report of three cases and review of the literature.

Castleman disease is a rare disorder characterized by benign lymph node hyperplasia involving lymphatic tissue in the neck, mediastinum, abdomen and other areas. Disease was described for the first time in 1956 by Castleman. The etiopathogenesis of the disease is unknown. The disorder can be classified into three histopathological types: hyalin-vascular, plasma-cell and mixed. We report three cases of the Castleman's disease (hyaline-vascular type) in three female patients with unilateral swelling of the neck. None of the patients developed any local or distant recurrence in postoperative follow-up.

Curcumin prevents oxidative renal damage induced by acetaminophen in rats.

Acetaminophen (APAP) can cause life-threatening renal damages and there is no specific treatment for APAP-induced renal damage. The aim of this study was to investigate the protective effects of curcumin (CMN) on APAP-induced nephrotoxicity. Nephrotoxicity was induced in male Wistar Albino rats by the administration of a single dose of 1000 mg/kg APAP intraperitoneally (i.p.). Some of these rats also received i.p. CMN (200mg/kg) at 30 min after the administration of APAP. Twenty-four hours after the administration of APAP, all the rats were sacrificed with a high dose of ketamine. Urea and creatinine levels were measured in the blood, and the levels of malondialdehyde (MDA) and glutathione (GSH), and antioxidant enzyme activity were determined in the renal tissue. Histopathological changes were studied. APAP administration caused elevated levels of renal MDA, and marked depletion of GSH levels and antioxidant enzyme activity, and deteriorated the renal functions as assessed by the increased plasma urea and creatinine levels as compared to control rats. CMN markedly reduced the elevated MDA levels, significantly increased the antioxidant enzyme activity and normalized the altered renal morphology in rats treated with APAP. CMN might be a potential candidate agent against APAP-induced nephrotoxicity, but further studies are required to identify this issue before clinical application becomes possible.

Comparison of tuberculin skin test and a specific T-cell-based test, T-Spot.TB, for the diagnosis of latent tuberculosis infection.

There is substantial evidence that the detection of T cells specific for the proteins ESAT-6 and CFP-10 using the ex vivo enzyme-linked immunospot technique is a marked improvement on the existing tuberculin skin test (TST). This new technique, which detects gamma-interferon-producing T cells, is now available as the commercial assay, T-Spot.TB. We compared the T-Spot.TB test with the TST for the diagnosis of latent tuberculosis infection (LTBI) in different groups of subjects. Significant accordance (85.7%) between the tests was found in subjects with active lung tuberculosis and in tuberculosis clinic and laboratory personnel (63.6%) but accordance was lowest and not significant amongst house contacts of tuberculosis patients (53.6%). We conclude that, in countries where vaccination is routinely performed, the T-Spot.TB test is a useful diagnostic test for LTBI in high-risk groups when carried out either together with the TST and/or to confirm the TST result.

Effects of zinc deficiency on oral and periodontal diseases in rats.

BACKGROUND AND OBJECTIVE: The aim of this study was to investigate the alterations of oral and periodontal tissues in zinc-deficient rats compared with control rats. MATERIAL AND METHODS: The study was carried out on 14 Sprague-Dawley rats, cessation of lactation on the 24th day after birth. Rats were randomly divided into two groups. Group I rats were fed with a zinc-deficient diet and group II rats were fed with a zinc-containing diet. At the end of the fourth week on experimental diets, alterations of the oral tissues in both groups were recorded. In addition, the gingival index (GI-Löe-Silness), plaque index (PI-Silness-Löe) and periodontal pocket depth scores were recorded in order to assess periodontal tissue health in the rats. Then, blood samples were taken and the serum zinc levels measured by atomic absorption spectrophotometry. At the end of the experiment, oral tissue samples were investigated by light microscopy. Finally, the results of the two groups were compared by using the Student's t-test. RESULTS: The effects of zinc deficiency were observed at 10-16 d in rats. Although body weight, body length and tail length were retarded in zinc-deficient rats, they were advanced in rats fed with a zinc-containing diet. The mean plaque index and gingival index for group II rats were significantly lower than for group I rats (p<0.001), but there was no significant difference regarding pocket depth between the two groups of rats (p>0.05). Aphthous ulcer was often seen in the study group, where it was observed on the alveolar mucosa with a high rate of 29.9%. According to histological findings, there was no difference related to the epithelial keratinization of the hard palate between the two groups. However, hyperkeratosis was found on the dorsal surface of the tongue in zinc-deficient rats. CONCLUSION: The findings indicated that oral health was better in group II rats (those fed with a zinc-containing diet) than in group I (zinc-deficient) rats. Hyperkeratinization was more prominent in zinc-deficient rats. We suggest that zinc deficiency is a potential risk factor for oral and periodontal diseases.