Screening the antifungal activities of monoterpenes and their isomers against Candida species.

AIM: Increasing the occurrence of non-albicans Candida species with intrinsic or acquired resistance to antifungals as well as the emergence of multidrug Candida species coupled with the limited antifungal agents challenges the treatment of candidiasis. Consequently, a class of secondary metabolites of plants exhibiting decent antifungal activity. Therefore, this study aimed to evaluate the antifungal potential of various monoterpenes including Carvone, Limonene, Pinene, Menthone, Menthol, Camphor, Thujone, Citronellol, and Piperitone against standard and clinical isolates of Candida. METHODS AND RESULTS: Minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of compounds were determined, using the broth Microdilution method based on M27-A3 protocol documented by clinical laboratory standard institute (CLSI). Amongst the tested monoterpenes, oxygenated terpenoids showed strong antifungal activity. Specifically, alcoholic terpenoids such as (±)-Citronellol possess more efficacy than the corresponding ketonic ones with MICs ranging from 0·03 to 2·00 µl ml-1 (0·16-10·80 mmol l-1 ). Among the examined yeasts, Candida tropicalis was the most susceptible species to (±)-Citronellol. Moreover, the examined monoterpenes successfully inhibited the growth of fluconazole-resistant Candida species. Moreover, statistical analysis showed no statistically significant difference between the (+) and (-) isomers, except for (±)-α-Pienene and (±) Menthone (ρ-value < 0·05). CONCLUSION: Among the tested monoterpenes, (±)-Citronellol was the most potent compounds followed by (+)-α-Pinene and Menthol. Considering the significant antifungal activity of the examined monoterpenes, they could be used in controlling or treating candidiasis. Those potent antifungal monoterpenes with GRAS status in addition to their pleasant taste and odour make them appropriate additive or preservative compounds in food and cosmetics products. Furthermore, these data might help researchers to predict EOs antifungal activities, after determining its constituents. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides new information about the antifungal activities of monoterpenes and their isomers presented widely in essential oils. Screening results against pathogenic yeasts confirm the correlation between the chemical structure of tested monoterpenes and their antifungal effects. The present findings might be helpful to anticipate the antifungal activity of essential oils.

Skewing of the genetic architecture at the ZMYM3 human-specific 5' UTR short tandem repeat in schizophrenia.

Differential expansion of a number of human short tandem repeats (STRs) at the critical core promoter and 5' untranslated region (UTR) support the hypothesis that at least some of these STRs may provide a selective advantage in human evolution. Following a genome-wide screen of all human protein-coding gene 5' UTRs based on the Ensembl database ( http://www.ensembl.org ), we previously reported that the longest STR in this interval is a (GA)32, which belongs to the X-linked zinc finger MYM-type containing 3 (ZMYM3) gene. In the present study, we analyzed the evolutionary implication of this region across evolution and examined the allele and genotype distribution of the "exceptionally long" STR by direct sequencing of 486 Iranian unrelated male subjects consisting of 196 cases of schizophrenia (SCZ) and 290 controls. We found that the ZMYM3 transcript containing the STR is human-specific (ENST00000373998.5). A significant allele variance difference was observed between the cases and controls (Levene's test for equality of variances F = 4.00, p < 0.03). In addition, six alleles were observed in the SCZ patients that were not detected in the control group ("disease-only" alleles) (mid p exact < 0.0003). Those alleles were at the extreme short and long ends of the allele distribution curve and composed 4% of the genotypes in the SCZ group. In conclusion, we found skewing of the genetic architecture at the ZMYM3 STR in SCZ. Further, we found a bell-shaped distribution of alleles and selection against alleles at the extreme ends of this STR. The ZMYM3 STR sets a prototype, the evolutionary course of which determines the range of alleles in a particular species. Extreme "disease-only" alleles and genotypes may change our perspective of adaptive evolution and complex disorders. The ZMYM3 gene "exceptionally long" STR should be sequenced in SCZ and other human-specific phenotypes/characteristics.

Evolving evidence on a link between the ZMYM3 exceptionally long GA-STR and human cognition.

The human X-linked zinc finger MYM-type protein 3 (ZMYM3) contains the longest GA-STR identified across protein-coding gene 5' UTR sequences, at 32-repeats. This exceptionally long GA-STR is located at a complex string of GA-STRs with a human-specific formula across the complex as follows: (GA)8-(GA)4-(GA)6-(GA)32 (ZMYM3-207 ENST00000373998.5). ZMYM3 was previously reported among the top three genes involved in the progression of late-onset Alzheimer's disease. Here we sequenced the ZMYM3 GA-STR complex in 750 human male subjects, consisting of late-onset neurocognitive disorder (NCD) as a clinical entity (n = 268) and matched controls (n = 482). We detected strict monomorphism of the GA-STR complex, except of the exceptionally long STR, which was architecturally skewed in respect of allele distribution between the NCD cases and controls [F (1, 50) = 12.283; p = 0.001]. Moreover, extreme alleles of this STR at 17, 20, 42, and 43 repeats were detected in seven NCD patients and not in the control group (Mid-P exact = 0.0003). A number of these alleles overlapped with alleles previously found in schizophrenia and bipolar disorder patients. In conclusion, we propose selective advantage for the exceptional length of the ZMYM3 GA-STR in human, and its link to a spectrum of diseases in which major cognition impairment is a predominant phenotype.

Stress among medical students of Gorgan (South East of Caspian Sea), Iran.

OBJECTIVE: This study aims to estimate the prevalence of psychological stress and association between the levels of stress and study variables among Gorgan medical students. MATERIALS AND METHODS: All three year medical students (129 basic sciences students) in Gorgan Faculty of Medicine, Golestan University of Medical Sciences, were asked to complete the Kessler 10 questionnaire. RESULTS: The findings showed mild, moderate and severe stress among 26.22%, 20.50% and 14.75% study subjects. 39.35% of medical students had no stress. There was statistically significant association between year of study and stress levels (p= 0.040). CONCLUSION: The results indicate that there is a decrease in the psychological health of first year medical students. Provided that stress management courses are organised by medical schools, when the students arrive, they will cope up with the stress in coming years. These courses may reduce the negative effects of stress on medical students. By providing such courses and reducing stress level, medical students may improve their medical education.

Interaction between Herpes Virus Infections and IL10 and Risk of Bone Marrow Suppression.

BACKGROUND: Syndrome of transient bone marrow suppression may result from various extra-hematological diseases, such as immunological deregulations, and viral infectious diseases secondarily affecting the function of hematopoietic stem cells. OBJECTIVE: To evaluate the pathogenic role of herpes viruses and their contraction with IL10 cytokine gene polymorphism, which can impair hematopoiesis in patients with transient bone marrow suppression. METHODS: In a cross-sectional study 30 patients who admitted to Namazi Hospital, affiliated to Shiraz University of Medical Sciences, with transient bone marrow suppression were recruited. Diagnosis of the transient bone marrow suppression was made by expert hematologists. A control group consisting of 100 healthy unrelated individuals was also included. One EDTA-treated blood sample was collected from each studied patients and plasma was isolated. The molecular prevalence of cytomegalovirus and HHV8 evaluated was evaluated using real-time and nested PCR protocols, respectively. The SNPs of the IL10 (rs 1800896-1082G/A) cytokine gene was evaluated by PCR-RFLP method. RESULTS: Cytomegalovirus and HHV8 infections were found in 2 and 3 of studied patients with transient bone marrow suppression. Significant higher frequency of IL10 G allele and GG genotype were found in HHV8-infected patients comparing to uninfected ones. Higher frequencies of A allele and AG and AA genotypes of IL10 were found in cytomegalovirus-uninfected patients comparing to infected ones, respectively. The significant higher frequencies of IL10 AA and AG genotypes were found in controls compared to bone marrow suppressed patients. CONCLUSION: IL10 genetic polymorphism might have determinative role in resistance to the cytomegalovirus, especially HHV8 infections, in patients with bone marrow suppression. Focus in new interaction between HHV8 infection and IL10 genetics in bone marrow suppressed patients should be completed by the analysis of the anti-herpes virus immunity in future studies.

Quantitation of ergosterol content and gene expression profile of ERG11 gene in fluconazole-resistant Candida albicans.

BACKGROUND AND PURPOSE: The frequency of opportunistic fungal infections in immunocompromised patients, especially by Candida species, has sharply increased in the last few decades. The objective of this study was to analyse the ergosterol content and gene expression profiling of clinical isolates of fluconazole-resistant Candida albicans. MATERIALS AND METHODS: Sixty clinical samples were identified and collected from immunocompromised patients, namely recurrent oral, vaginal, and cutaneous candidiasis, during 2015-16. Antifungal susceptibility testing of fluconazole against clinical Candida species was performed according to Clinical and Laboratory Standards Institute guidelines. Ergosterol content and gene expression profiling of sterol 14α-demethylase (ERG11) gene in fluconazole-susceptible and -resistant C. albicans were investigated. RESULTS: The specimens consisted of C. albicans (46.67%), Candida krusei (41.67%), and Candida tropicalis (11.67%). All the isolates were resistant to fluconazole. No significant reduction was noted in total cellular ergosterol content in comparison with untreated controls in terms of fluconazole-resistant C. albicans. The expressionlevel of ERG11 gene was down-regulated in fluconazole-susceptible C. albicans. Eventually, the expression pattern of ERG11 gene revealed no significant changes in fluconazole-resistant isolates compared to untreated controls. The results revealed no significant differences between fluconazole-susceptible and -resistant C. albicans sequences by comparison with ERG11 reference sequence. CONCLUSION: Our findings provide an insight into the mechanism of fluconazole resistance in C. albicans. The mechanisms proposed for clinical isolates of fluconazole-resistant C. albicans are alteration in sterol biosynthesis, analysis of expression level of ERG11 gene, and analysis of gene sequences. Nonetheless, further studies are imperative to find molecular mechanisms that could be targeted to control fluconazole resistance.

Association of TLR4 polymorphisms and polyomavirus BK infection in liver transplant patients.

Toll-like receptors (TLRs) may have a role in orchestrating the immune responses against polyomavirus BKV and also may influence liver transplant outcomes. However, the clinical relevance of this experimental observation has not been examined. Improving knowledge regarding details of genetic source of TLR polymorphisms can promote new therapeutic strategies to inhibit virus related clinical disorders in post-liver transplantation. Therefore, the Asp299Gly and Thr399Ile TLR4 polymorphisms were evaluated in liver transplanted patients with and without polyomavirus BK infection. In a cross sectional study, 144 liver transplant patients received allograft at Transplant Center of Namazi Hospital affiliated to Shiraz University of Medical Sciences who were recruited between years: 2014- 2015. Patients were followed for the graft outcome and acute rejection episode(s) and divided into two groups based on experiencing acute rejection or not. The genomic DNA of polyomavirus BK was diagnosed in studied patient using qualitative nested- PCR technique. Analysis of TLR4 gene polymorphisms were analyzed using PCR-RFLP protocols. The polyomavirus BK infection was found in 15 of 144 (10.4%) liver transplanted patients. A total of 14 of 15 (93.3%) and all of polyomavirus BK infected patients have been shown to be homozygous wild type AA genotype of TLR4-Asp299Gly (A896G) and CC genotype of TLR4- Thr399Ile (C1196T) polymorphisms. Homozygous mutant GG genotype of Asp299Gly (A896G) was found in 3 (2.1%) of the studied patients. Homozygous mutated TT genotype of Thr399Ile (C1196T) was found in only 5 (3.5%) of the liver recipients. There were no significant differences between homozygous wild type genotypes of studied TLR4 SNPs for liver transplant patients with or without polyomavirus BK infections. Significant association was also not found between homozygous mutated genotype of TLR4 SNPs for patients experiencing rejection episodes. However further completed studies on larger population and with longer follow-up are needed to confirm these results.

A comparison between tourniquet application and epinephrine injection for hemostasis during hypospadias surgery: The effect on bleeding and postoperative outcome.

OBJECTIVE: To compare tourniquet application and epinephrine injection for hemostasis during hypospadias surgery in terms of bleeding and complications. METHODS: Between April 2013 and September 2014, patients who were admitted for hypospadias repair were divided by random allocation into either a diluted epinephrine injection (DE) group or tourniquet application (T) group for hemostasis during the procedure. In the T group, a rubber band was applied at the base of the penis after skin dissection, and in the DE group, epinephrine 1/100,000 was injected along the incision lines. The patients' ages, urethral defect lengths, pre-operative, intraoperative and postoperative variables were compared between the two groups. RESULTS: A total of 70 patients (35 in each group) were enrolled into the study. The mean ages and preoperative variables were not significantly different. Average blood loss was 23.51 ± 15.36 cc in the tourniquet group and 15.99 ± 10.00 cc in the epinephrine group, and was significantly higher in tourniquet group (P = 0.022); however, the mean operative time was not significantly different. Postoperative complications, described as Clavian classification, were reported in eight patients (23%) in the T group and 10 patients (30%) in the DE group, which was not significantly different. CONCLUSION: Epinephrine injections minimize operative bleeding without significant harmful effects on postoperative outcomes; therefore, it could be considered to be a safe and effective method for preparation of a bloodless field during hypospadias surgery.

Physical mapping of chromosomes using unique probes.

The goal of physical mapping of the genome is to reconstruct a strand of DNA given a collection of overlapping fragments, or clones, from the strand. We present several algorithms to infer how the clones overlap, given data about each clone. We focus on data used to map human chromosomes 21 and Y, in which relatively short substrings, or probes, are extracted from the ends of clones. The substrings are long enough to be unique with high probability. The data we are given is an incidence matrix of clones and probes. In the absence of error, the correct placement can be found easily using a PQ-tree. The data are never free from error, however, and algorithms are differentiated by their performance in the presence of errors. We approach errors from two angles: by detecting and removing them, and by using algorithms that are robust in the presence of errors. We have also developed a strategy to recover noiseless data through an interactive process that detects anomalies in the data and retests questionable entries in the incidence matrix of clones and probes. We evaluate the effectiveness of our algorithms empirically, using simulated data as well as real data from human chromosome 21.

Effect of uterine contraction and amniotomy on fetal cardiotocograph.

Both uterine contractions and artificial rupture of fetal membranes (amniotomy) are important events during delivery phase, this study was planned to determine possible effects of both events on fetal heart rate using nonstress test method. Sixty term pregnant women admitted for termination of pregnancy were selected. Primary nonstress test was performed, then nonstress test was done after active uterine contractions. After the amniotomy the last test was done. All results were statistically analyzed. Out of 60 term pregnant women, 51 (85%) patients reported acceleration for primary nonstress test. After uterine contractions, acceleration was seen in 46 (76.7%) patients, this happened in 40 (66.7%) cases after amniotomy. There was no deceleration after primary nonstress test. However, deceleration reported after uterine contractions and amniotomy in five (8.3%) and two (3.3%) cases, respectively. The 56 (93.3%) patients showed variability in primary nonstress test; variability following the uterine contractions and amniotomy was seen in 58 (96.7%) and 56 (93.3%) of subjects, respectively. None of them were statistically significant (p > 0.05). Mean fetal heart rate baseline in primary Nonstress test was (141.00 +/- 9.35), this was (140.50 +/-10.51) following uterine contractions and (143.08 +/- 11.97) after amniotomy. Baseline fetal heart rate reduction, statistically significant, was seen following uterine contractions (r = +0.28, p = 0.02). This means reductions in lower baseline fetal heart rates will be more outstanding. Uterine contractions and amniotomy had no correlation with presence or absence of variability, acceleration and deceleration.

Use of leisure time in cardiovascular patients in Gorgan (south east of Caspian Sea).

The aim of this study was to compare activity patterns and leisure time between matched groups of patients with cardiovascular disease and individuals without a heart disease. The study included 100 patients recruited from those referred to cardiology department of 5th Azar General Hospital of Golestan University of Medical Sciences in Gorgan (South East of Caspian Sea) and 100 matched control subjects during the period 2007-2008. Odds ratios (OR), together with 95% confidence intervals (95% CI), were calculated using logistic regression, as estimates of relative risks. Listening to music OR = 8.800 (95% CI: 2.717-28.499, p<0.05), meditation OR = 6.111 (95% CI; 2.616-14.274, p<0.05) were independent risk factors. Subjects who performed 2 h per week and 2-4 h per week physical activity, the odds ratios were 0.038 (95% CI: 0.012-0.124, p<0.05) and 0.079, (95% CI: 0.024-0.260, p<0.05), respectively. Low physical activity and use of long time relaxation are associated with cardiovascular disease in these patients. Regular participation in physical activity such as walking 2 h per week and 2-4 h per week, are associated with reduced risk of cardiovascular disease. This study suggests the importance of both leisure-time physical activity and sedentary behaviors in the prevention of CVD.