Molecular identification, virulence factors and antifungal susceptibility patterns of Candida parapsilosis complex species isolated from clinical samples.

AIMS: The aim of this study was to identify C. parapsilosis complex strains isolated from various clinical samples by sequence analysis and to investigate whether there are any differences between the species in terms of virulence factors and antifungal susceptibility. METHODS: The study included a total of 42 isolates identified as C. parapsilosis complex based on the color they formed in chromogenic medium, colony morphology, and microscopic appearance in Corn Meal-Tween 80 Agar and they were confirmed with API 20 C AUX. For the DNA sequence analysis of clinical isolates, V9G forward and LS reverse primers were used as well as internal transcribed spacers (ITS1 and ITS4). Biofilm formation, esterase, phospholipase, and protease activities were evaluated as virulence factors. Antifungal susceptibility was investigated via colorimetric microdilution method. RESULTS: A total of 75 non-C. albicans isolates were obtained from various clinical samples between 2016 and 2017 in a Turkish Tertiary Care Hospital. Of them, 42 were identified as members of the C. parapsilosis complex. Of the 42 strains, 41 were identified as C. parapsilosis sensu stricto (CpSS), while only one was identified as C. orthopsilosis. Of the CpSS strains, 31 (75.6%) were biofilm-positive, six (14.6%) were esterase-positive, nine (21.9%) were positive for phospholipase activity, and 31 (75.6%) were positive for protease formation, whereas all virulence factors of C. orthopsilosis strain were found to be negative. All CpSS strains were found susceptible to amphotericin B, echinocandins, and flucytosine. CONCLUSIONS: It has been concluded that the use of molecular methods to identify CpSS would not be effective in routine laboratory practices as it is the most commonly isolated species from the C. parapsilosis complex and there are no significant differences between species in terms of antifungal susceptibility.

Can serums be replaced by Mueller-Hinton agar in germ tube test?

BACKGROUND: The germ tube test (GTT) is inexpensive, easy, and well-defined test that differentiates Candida albicans (excluding Candida dubliniensis and Candida africana) from other species. The aim of this study was to evaluate various serums (i.e., human, rabbit, horse, and fetal bovine serum) used in the GTT and Mueller-Hinton agar (MHA). MATERIALS AND METHODS: Fifty species isolated from various clinical samples that were defined as C. albicans by both conventional and DNA sequence analysis methods were included in the study. One to two colonies of C. albicans were mixed into 0.5-1 ml of fetal bovine serum, horse serum, rabbit serum, and human serum. Serums and MHA were incubated at 37°C for GTT. They were removed from the incubator and evaluated after 30 min, 1 h, 2 h, and 3 h of incubation. The GTT was accepted to be positive only if germ tube was 1/2 the width and 3 times the length of the parent yeast cell and with no constriction at the point of origin. RESULTS: When the use of serums and MHA for GTT was statistically evaluated, according to the positive scoring, the best results were obtained with MHA and with rabbit, horse, and fetal bovine serum, respectively. The best definition over time statistically was the third hour. CONCLUSION: It is suggested that inexpensive MHA is a fast, appropriate, and reliable medium for the probable diagnosis of GTT and C. albicans; however, additional studies are still needed to define other Candida species.

Change in the ovarian environment after hysterectomy with bilateral salpingectomy: is it the technique or surgery itself?

OBJECTIVE: To compare the effects of total laparoscopic hysterectomy with bilateral salpingectomy (TLH-BS) and total abdominal hysterectomy with bilateral salpingectomy (TAH-BS) on ovarian function among women of reproductive age. STUDY DESIGN: One hundred and three patients with a diagnosis of benign uterine disorder were divided into two groups in this prospective longitudinal study. Patients who had never had sexual intercourse and patients with uterovaginal disproportion underwent TAH-BS (n=57), and the remaining patients (n=46) underwent TLH-BS. Ovarian function was assessed before and 6 months after surgery; ovarian volume was assessed by gray-scale ultrasonography, and levels of serum follicle-stimulating hormone (FSH), luteinizing hormone (LH), oestradiol (E2), anti-Mullerian hormone (AMH) and inhibin B were measured. RESULTS: Postoperative serum FSH, LH and inhibin B decreased significantly in both groups. Postoperative serum E2 did not change significantly. Postoperative serum AMH and ovarian volume decreased significantly in the TAH-BS group (p=0.016 and p<0.001, respectively), but not in the TLH-BS group. Significant differences were observed between the TLH-BS and TAH-BS groups with respect to change in FSH (p=0.012) and ovarian volume (p=0.001); between-group differences were not significant for changes in AMH and inhibin B. CONCLUSIONS: Although serum AMH did not change significantly in patients who underwent TLH-BS, ovarian aging commenced following both surgical procedures.

Clinical significance of maternal serum vascular endothelial growth factor (VEGF) level in idiopathic recurrent pregnancy loss.

OBJECTIVE: The aim of this study was to determine whether maternal serum vascular endothelial growth factor (VEGF) levels are associated with unexplained recurrent pregnancy losses (RPLs). PATIENTS AND METHODS: Twenty-one pregnant women with idiopathic RPLs who were selected from 47 cases with RPLs were compared with age-matched 24 control participants. Transvaginal obstetric ultrasonographies were performed and maternal serum samples were collected between 5th and 10th gestational weeks to evaluate serum VEGF and progesterone (P4) concentrations. Enzyme-linked immunosorbent assay technique was used in measurements of VEGF and P4. RESULTS: Prevalence of idiopathic cases among all RPLs was 44.7%. Median serum VEGF value was found statistically higher in RPL group when compared to control group (210.33 ± 108.23 pg/ml vs. 123.91 ± 18.8 pg/ml, respectively). There was no statistical difference between the median values of serum P4 levels in idiopathic RPL group and the control group (19.53 ± 5.79 ng/ml and 20.08 ± 7.85 ng/ml, respectively). Serum VEGF levels did not differ significantly with regard to gestational age within the RPL and control groups (p = 0.72 and p = 0.89, respectively). A positive correlation was found between VEGF levels and the patients' age within RPL group (r = 0.515). CONCLUSIONS: Serum VEGF levels are independent by the gestational age. Serum VEGF concentrations correlate positively with maternal age. Increased maternal age, especially maternal age over 35 years, is related to elevated serum VEGF concentration. Increased maternal serum VEGF concentration is related to recurrent pregnancy loss.

The possible role of serum leptin in preeclampsia.

BACKGROUND: It is theorized that adipokines play a critical role in the pathophysiology of preeclampsia, particularly with their pro-inflammatory and inflammatory features. AIM: To investigate serum leptin levels in pregnancies complicated with preeclampsia and severe preeclampsia. MATERIALS AND METHODS: Maternal serum leptin levels were analyzed by solid phase enzyme amplified sensitivity immunoassay (EASIA) method in 23 patients with mild preeclampsia, 29 patients with severe preeclampsia, and 28 healthy pregnant controls. RESULTS: Mean serum leptin levels did not differ statistically between patients with mild preeclampsia, severe preeclampsia, and the controls (10.77 ng/ml, 13.40 ng/ml, and 8.43 ng/ml, respectively). Also, there was no relationship between serum leptin levels and the gestational ages of the participants. DISCUSSION: Serum leptin levels are not associated with preeclampsia. Leptin measurements are not affected with the gestational age. The role of leptin in the pathophysiology of preeclampsia should be evaluated cautiously.

Detecting coagulability status by thromboelastography in women with the history of preeclampsia and inherited thrombophilia.

OBJECTIVE: To assign tendency to thrombosis in patients with preeclampsia and inherited thrombophilia using thromboelastography (TEG), and therefore to evaluate possible relationship between thrombophilia and preeclampsia. MATERIALS AND METHODS: Kinetics of clot formation was assessed with TEG analyzer in 49 patients with severe preeclampsia, 54 cases with previous diagnosis of inherited thrombophilia, and 31 controls. RESULTS: 'r, 'k', TMA, coagulation index (CI) parameters were found statistically discrete between patients with inherited thrombophilia and controls. The difference between preeclampsia and control groups was not statistically significant. The difference in a angle was statistically significant between thrombophilics and preeclamptics (p = 0.01), and between thrombophilics and controls (p = 0.004). CI was found statistically lower in thrombophilia group than control group (p = 0.006). Particularly, clot lysis time (CLT) was measured to shorten in preeclampsia when compared with controls and patients with thrombophilia (p = 0.032, p = 0.028, respectively). CONCLUSIONS: Not only the inherited thrombophilia group but also preeclampsia group demonstrated elongated clot initiation patterns when compared to the controls. Moreover, apart from the patients with inherited thrombophilia, preeclamptics exposed shorter CLT values indicating a possible increment in clot turn over, which eventually results in increased depletion of coagulation substrates, and thus, increased frequencies of oxidative cycle injury.

Investigation of possible virulence factors in Candida strains isolated from blood cultures.

BACKGROUND: The Candida species, which are one of the most common causes of nosocomial bloodstream infections, present with high mortality and morbidity rates. This study aims to investigate the production of esterase, phospholipase, proteinase, and biofilm formation ability of the Candida strains isolated from the blood cultures. MATERIALS AND METHODS: Between June 2011 and July 2012, the Candida strains, which were isolated from blood cultures of a total of 50 patients, were studied. The esterase activity was analyzed in the Tween-80 agar, while phospholipase activity was studied in the egg yolk agar. The proteinase activity and biofilm formation were identified by using the petri dish method and microplate method, respectively. RESULTS: Of 50 specimens obtained from individual patients, 17 (34%) were identified as C. albicans, 14 (28%) as C. glabrata, 9 (18%) as C. parapsilosis, 5 (10%) as C. krusei, 4 (8%) as C. kefyr, and 1 (2%) as C. tropicalis. The rate of proteinase, phospholipase, and esterase positivity was higher in the C. albicans isolates. Biofilm formation was the highest in the C. parapsilosis strains. CONCLUSIONS: Higher rate of virulence factors in the most commonly isolated Candida species than other species indicates that these virulence factors play a crucial role in the pathogenesis.

Glutathione s-transferase m1 and t1 gene polymorphisms are not associated with increased risk of gestational diabetes mellitus development.

AIM: The aim of this study was to investigate whether the glutathione S-transferase M1 (GSTM1) and T1 (GSTT1) gene polymorphisms contributed to development of gestational diabetes mellitus (GDM). SUBJECTS AND METHODS: Fifty women with diagnosis of GDM and 50 control individuals without GDM or altered glucose intolerance during their pregnancy were enrolled in the study. Multiplex polimerase chain reaction-restriction fragment length polymorphism method was applied to determine the GSTM1 and GSTT1 gene polymorphisms. Genotypes were determined according to bands detected with the agarose gel electrophoresis. RESULTS: The difference in the frequencies of GSTM1 null genotypes between GDM and control groups was not statistically significant (60% and 54%, respectively). There was no statistically significant difference between GDM and control groups with respect to GSTT1 null genotype rates (22% and 20%, respectively).There was no statistically significant difference between GDM and control groups with respect to GSTT1 null genotype rates (22% and 20%, respectively). CONCLUSION: This study shows no association between GST gene polymorphisms and GDM.

Maternal adiponectin and visfatin concentrations in normal and complicated pregnancies.

OBJECTIVE: To evaluate the role of adiponectin and visfatin in the pathophysiology of pre-eclampsia (PE) and how their concentrations correlate with the severity of the disease and neonatal outcomes. STUDY DESIGN: A prospective case-control study was carried out in 52 preeclamptic and 28 healthy pregnant women during the third trimester. The maternal plasma concentrations of adiponectin and visfatin were determined. Neonatal outcomes were also recorded. RESULTS: Mean maternal plasma adiponectin concentrations in healthy pregnant women did not differ significantly from those of mild PE and severe PE groups. The plasma adiponectin levels of PE patients with small for gestational age (SGA) and those without SGA did not differ significantly, but the median plasma visfatin concentration of patients with SGA fetus was significantly higher if the patient was preeclamptic (p = 0.036). CONCLUSION: The severity of preeclampsia did not change the plasma levels of adiponectin and visfatin, but the median plasma visfatin concentration of patients with SGA fetuses were significantly higher if the patient was preeclamptic. Altered levels of adipocytokines strongly imply that the regulation of adipocytokines in PE is different and more complex compared to that in healthy pregnancy.

Polymorphisms of glutathione-s-transferase M1, T1, and P1 genes in endometrial carcinoma.

OBJECTIVE: To investigate the polymorphism rates and possible roles of glutathione-s-transferase M1, T1, and P1 gene polymorphisms in the predisposition to endometrial cancer in Caucasian women. MATERIALS AND METHODS: Serum samples and medical records were collected from 53 Caucasian women with newly diagnosed endometrial cancer and 67 women of the same race without any known history of cancer. Multiplex polymerase chain reaction (PCR) analysis was used to assess glutathione-s-transferase M1 (GSTM1) and T1 gene polymorphisms. Polymerase chain reaction - restriction fragment length polymorphism (PCR-RFLP) method was used in salvage of GSTP1 gene polymorphism. RESULTS: Frequencies of GSTM1 and GSTT1 null genotypes were not significantly different between the controls and patients with endometrial cancer (56.7% vs 52.8%, p = 0.671; 32.8% vs 26.4%, p = 0.574, respectively). The authors were not able to demonstrate any association between neither GSTP1 genotypes nor allele frequencies and endometrial carcinoma in the population studied (p = 0.310, p = 0.318, respectively). Moreover, no significant association could be demonstrated with GSTM1 and GSTT1 polymorphisms and clinical stages of endometrial cancer. Nevertheless, there was a significant difference between the frequencies of GSTP1 AA and GG genotypes in relation to Stage I disease when compared with advanced stages of endometrial carcinoma (p = 0.03). In addition, no association was found between polymorphisms of GST suspergene family and non-endometrioid type endometrial carcinomas. CONCLUSION: These results suggest that GSTT1, GSTM1, and GSTP1 polymorphisms are not associated with endometrial cancer in the Caucasian population.

Polymorphisms in angiotensin-converting enzyme and glutathione s-transferase genes in Turkish population and risk for preeclampsia.

AIMS: This study was conducted to investigate whether insertion/deletion (I/D) polymorphism of angiotensin-converting enzyme (ACE) gene and polymorphisms in glutathione S-transferase (GST) M1 and T1 genes are associated with increased risk for preeclampsia. MATERIALS AND METHODS: Sixty-three patients with hypertensive disorder of pregnancy and 85 controls were evaluated in a prospective case-control study. All subjects were genotyped by polymerase chain reaction (PCR) followed by agarose gel electrophoresis. RESULTS: Allele frequencies of ACE gene I/D polymorphism were found significantly different between preeclampsia and the control groups (p = 0.001). Differences in genotype frequencies of ACE gene I/D polymorphism between the two groups were statistically significant (p = 0.004). Individuals homozygous for D allele were more likely to develop preeclampsia (OR = 2.29; 95% CI, 1.39-3.79), whereas heterozygous individuals were not at increased risk (OR = 0.92; 95% CI, 0.56-1.49), compared to individuals homozygous for I allele. The differences in frequencies of functional and null alleles of GSTM1 and GSTT1 genes between the two groups were not significant (p = 0.46 and p = 0.44, respectively). CONCLUSION: ACE gene DD genotype was found to be associated with increased risk of preeclampsia development, whereas the authors did not find any significant relationship with polymorphisms of the GSTM1 and GSTT1 genes and preeclampsia.

Vascular endothelial growth factor gene 1154 G/A, 2578 C/A, 460 C/T, 936 C/T polymorphisms and association with recurrent pregnancy losses.

Vascular endothelial growth factor (VEGF) regulates endothelial cell proliferation, migration and differentiation. VEGF plays a critical role in angiogenesis during placenta formation. We investigated whether VEGF gene polymorphisms are associated with recurrent pregnancy loss. Thirty-eight women with recurrent pregnancy loss and 30 control women with live-born children were recruited from 2010 to 2011 in the region of Bursa, Turkey. VEGF gene polymorphisms were assessed with PCR-RFLP analysis of DNA samples obtained from leukocytes. DNA fragments were investigated by using appropriate primers. SNP scanning was performed using MnII, BgIII, BshI2361, Hsp92II restriction enzymes for 1154 G/A, 2578 C/A, 460 C/T, and 936 C/T polymorphisms, respectively. The frequencies of 2578 C/A, 460 C/T, 936 C/T polymorphisms were not significantly different between the controls and women with recurrent pregnancy loss. However, the prevalence of the 1154 G/A polymorphism A/A genotype was significantly higher in the recurrent pregnancy loss group (23.7 vs 3.4%). One of the four common polymorphisms of the VEGF gene was found to be more frequent in women with recurrent pregnancy loss. It is possible that disruption of VEGF function and placental angiogenesis can contribute to pregnancy loss in women with recurrent pregnancy loss.