Genotype and clinical phenotype of children with Marfan syndrome in Southeastern Anatolia.

The cardinal phenotypic hallmarks of Marfan syndrome (MFS) include cardiac, ocular, and skeletal abnormalities. Since the clinical phenotype of MFS is highly heterogeneous, with certain symptoms appearing as children age, the diagnostic process and establishing a genotype-phenotype association in childhood MFS can be challenging. The lack of sufficient childhood studies also makes it difficult to interpret the subject. This study aims to evaluate the relationship between clinical symptoms used as diagnostic criteria and FBN1 variations in children with MFS. This study investigated the relationships between genotypes and phenotypes in 131 children suspected of having Marfan syndrome (MFS). Diagnosis of MFS was made according to the revised Ghent nosology. FBN1 variants were categorized based on exon regions, type of variant, and pathogenicity classes. These FBN1 variants were then correlated with the clinical manifestations including cardiovascular, ocular, facial, and skeletal abnormalities. Out of the children, 43 were diagnosed with MFS. FBN1 variant was identified in 32 (74.4%) of the MFS children. MFS diagnosis could not be made in five (15.6%) FBN1 variant-positive children. The most common cardinal finding is cardiac anomalies n = 38 (88.3%). The most common FBN1 pathogenic variant was c.1786 T > C/p.Cys596Arg n = 4 (12.5%). The distribution of pathogenic variants was as follows: 29 (90.6%) missense, 2 (6.3%) frameshift, and 1 (3.1%) nonsense. The numbers of AD and EL of the variant-positive children were 16 (50%) and 14 (43.7%), respectively. Ocular abnormalities were more common in children with FBN1-positive MFS (p = 0.009). There was no difference in the number of cardiac abnormalities between FBN1-positive and FBN1-negative MFS patients (p = 0.139).   Conclusion: This study examines the relationship between FBN1 variants and clinical features used as diagnostic criteria in MFS children. The findings emphasize the importance of long-term monitoring of heterogeneous clinical phenotypes and bioinformatic reanalysis in determining the genotype-phenotype relationship in children, as MFS symptoms can vary with age. What is Known: • Marfan syndrome has highly variable phenotypic heterogeneity. • The genotype-phenotype relationship in childhood Marfan syndrome is not clear enough due to the variation in the time of onset of the findings. What is New: • This article provides regional data for the field of research on genotype-phenotype relationships in childhood Marfan syndrome. • Long-term follow-up of clinical findings and bioinformatics reanalysis is an important requirement for a well-established genotype-phenotype relationship in childhood Marfan syndrome.

lncRNA NORAD, soluble ICAM1 and their correlations may be related to the regulation of the tumor immune microenvironment in laryngeal squamous cell carcinoma (LSCC).

NORAD, non-coding RNA activated by DNA damage, is a Long non-coding RNA (lncRNA) transcript that modulates genome stability and has been reported to be dysregulated in different cancers. Although it has been reported to be upregulated in tumor cells mostly for solid organ cancers, it has also been reported to be downregulated in some cancers. Although the pathophysiological mechanism is not fully understood, a negative correlation between NORAD and intercellular cell adhesion molecule-1 (ICAM-1) has been shown in experimental models, but this situation has not been evaluated in terms of cancer. We aimed to evaluate the potential roles of these two biomarker candidates together and separately in the clinicopathological axis in Laryngeal squamous cell carcinoma (LSCC) in a case-control study setting. The interactions of NORAD and ICAM1 at the RNA level were evaluated interactively by the RIblast program. sICAM1 (soluble intercellular cell adhesion molecule-1) levels were determined by ELISA in one hundred and five individuals (forty-four LSCC, sixty-one control) and lncRNA NORAD expression in eighty-eight tissues (forty-four LSCC tumors, forty-four tumor-free surrounding tissues) was determined by Real-time PCR. While the energy treesholud was - 16 kcal/mol between NORAD and ICAM1, the total energy was 176.33 kcal/mol, and 9 base pair pairings from 4 critical points were detected. NORAD expression level was found to be higher in tumor surrounding tissue compared to tumor tissue, and sICAM1 was higher in the control group compared to LSCC (p = 0.004; p = 0.02). NORAD discreminte tumor surrounding tissue from tumor (AUC: 0.674; optimal sensitivity:87.50%; optimal specificity 54.55%; cut-off point as >1.58 fold change; P = 0.034). The sICAM1 level was found to be higher in the control (494,814 ± 93.64 ng/L) than LSCC (432.95 ± 93.64 ng/L) (p = 0.02). sICAM1 discreminte control group from LSCC (AUC: 0.624; optimal sensitivity 68,85%; optimal specificity 61,36%; cut-off point ≤115,0 ng/L; (p = 0.033). A very strong negative correlation was found between NORAD expression and patients' sICAM1 levels (r = -.967; n = 44; p = 0.033). sICAM1 levels were found to be 1.63 times higher in NORAD downregulated subjects compared to upregulated ones (p = 0.031). NORAD was 3.63 times higher in those with alcohol use, and sICAM 1 was 5.77 times higher in those without distant organ metastasis (p = 0.043; 0.004). The increased NORAD expression in the tumor microenvironment in LSCC, the activation of T cells via TCR signaling, and the decrease of sICAM in the control group in correlation with NORAD suggests that ICAM1 may be needed as a membrane protein in the tumor microenvironment. NORAD and ICAM1 may be functionally related to tumor microenvironment and immune control in LSCC.

Investigation of Long Non-coding RNAs H19 and LINC00675 in Colorectal Cancers in Terms of Histopathological Features and Correlations With Plasma Markers.

BACKGROUND/AIM: Functional and bioinformatic studies provide strong evidence that long non-coding RNAs (lncRNAs) can alter the molecular mechanisms of cancer through their interactions with DNA, RNAs, and proteins. This study aimed to evaluate the role of H19 and LINC00675 lncRNAs in colorectal cancers (CRCs) in terms of clinicopathological features. MATERIALS AND METHODS: Tumor and tumor-free surrounding tissue samples were obtained from 51 CRC cases. Total RNA isolation and cDNA synthesis were performed. qPCR was performed using the TaqMan non-coding lncRNA assay specific for H19 and LINC00675. Preoperative levels of plasma markers, lncRNA expression, and clinicopathological characteristics of the cases were evaluated statistically. RESULTS: Expression of H19 in tumor tissue was found to be 2.11 times higher than that of tumor-free surrounding tissue (p<0.001). LINC00675 levels were found to be approximately three times higher in colon tumors than tumors with rectal involvement (p=0.019). There was a correlation between H19 expression and creatinine (r=0.408; p=0.003). In addition, correlations were detected between LINC00675 with albumin (r=0.303; p=0.03), and between LINC00675 with globulin (r=0.332; p=0.02). CONCLUSION: H19 is a candidate biomarker that can be evaluated in terms of prognosis and antineoplastic treatment response, while LINC00675 may be an important marker of the microenvironment of advanced stage tumors, especially in tumors with rectal involvement.

Immunological interpretation of minipuberty: Minipuberty as the driving force of sexual dimorphism in the immune response.

BACKGROUND/OBJECTIVE: Although the physiology of minipuberty is well established, it is not fully explained why it occurs. It has been suggested that minipuberty contributes to the development of reproductive organs, somatic growth, cognitive/behavioural and sex-specific brain development. Given the well-known trade-off between the reproductive/endocrine and immune systems in adults, the immunological approach to minipuberty, which is characterized by transient activation of the hypothalamo-pituitary gonadal (HPG) axis, seems to be ignored. This study focused on the relationship of the lymphocyte subsets with gonadotrophin and sex hormones during the minipuberty. MATERIALS AND METHODS: A total of 121 newborns (58 male) were included in this cross-sectional study. The hormone and lymphocyte subsets studied were as follow: follicle-stimulating hormone (FSH), luteinizing hormone (LH) estradiol (E), testosterone (T), CD19, CD16/56, CD3, CD3/CD4, CD3/CD8 and HLA-DR as lymphocyte activation marker. RESULTS: The mean FSH levels were higher in females (15.15 ± 10.12 mIU/ml vs, 2.61 ± 1.74 mIU/ml) and LH in males (5.80 ± 2.51 mIU/ml vs. 1.91 ± 12.89 mIU/ml) (P < .001 for each). The mean percentages of the CD3/CD4 levels were higher in females (54.61 ± 6.70% vs. 51.17 ± 6.77%) and CD3/CD8 in males (21.49 ± 4.82% vs. 17.31 ± 3.66%) (P < .001 for each). In the females, the mean FSH levels negatively correlated with CD3/CD4 (rFSH-CD3/CD4  = -0.423, P = .001) and positively correlated with CD3/CD8 (rFSH-CD3/CD8  = 0.311, P = .013). In the males, LH positively correlated with CD3/CD4 (rLH-CD3/CD4  = 0.406, P < .001) and negatively correlated with CD3/CD8 (rLH-CD3/CD8  = -0.486, P < .001). CONCLUSION: This study showed that the mean CD3/CD4 levels were higher in female and CD3/CD8 in male newborns, indicating that there was a sexual dimorphism in favour of immunostimulant in females and immunosuppressor components of immune response in males during the minipuberty. These interactions point to sex-specific trade-off between reproductive/endocrine and immune systems, which it reflects the an investment favouring in the reproductive system against the immune response during minipuberty, which is critical period for adult fertility, especially in males.

Immunophenotypic analysis of lymphocyte subsets in newborns with biotinidase deficiency.

BACKGROUND/AIM: Biotin is a vital micronutrient that plays a role in metabolic homeostasis and the regulation of innate and adaptive immune system functions. Biotinidase deficiency (BTD) leads to impairment in biotin-dependent immune functions. This study focused on immunophenotypic analysis of lymphocyte subsets in newborns with BTD. PATIENTS AND METHODS: A total of 181 (95 female and 86 male; 114 had BTD and 67 were healthy) newborns underwent biotinidase enzyme activity, molecular and lymphocyte immunophenotyping analyses. BTD is classified into four biochemical phenotypes: profound, partial, heterozygous and normal. The following lymphocyte subsets were studied in all participants: total B lymphocyte (CD19), total T lymphocyte (CD3), helper T lymphocyte (CD3/CD4), cytotoxic T lymphocyte (CTL) (CD3/CD8), natural killer T lymphocyte (CD16/56) and a T-lymphocyte activation marker (HLA-DR). RESULTS: The percentages of lymphocyte subsets were similar in newborns with and without BTD. In all newborns with BTD, the mean CD3/CD4 levels were higher in females, while the CD3/CD8 levels were higher in males (P < .001 for each). In female and male newborns, the CD3/CD4 levels were 53.83 ± 9.46 and 16.82 ± 5.19, respectively, and the CD3/CD8 levels were 48.80 ± 8.65 and 21.48 ± 6.02, respectively. A moderate negative correlation was found between CD3/CD4 and CD3/CD8 in female and male newborns (rfemale  = -0.488, rmale  = -0.574, P < .001). CONCLUSION: This study showed that although there were no differences in the lymphocyte subsets in newborns with BTD, the CD3/CD4 levels were higher in females, and the CD3/CD8 levels were higher in males. In addition, there was a negative correlation between the CD3/CD4 and CD3/CD8 levels in both genders. Although these results indicate sexual dimorphism between CD3/CD4 and CD3/CD8 levels, whether this dissociation is unique to BTD in newborns is not fully clear.

Clinical and laboratory clues of maturity-onset diabetes of the young and determination of association with molecular diagnosis.

BACKGROUND/AIM: Maturity-onset diabetes of the young (MODY) is often misdiagnosed as other types of diabetes because it is overlooked due to atypical clinical presentations. This study aims to reveal the clinical and laboratory clues and examine their compatibility with MODY genotypes. METHODS: Participants consisted of 230 children with atypical presentations for type1(T1DM) and type2 diabetes mellitus (T2DM). MODY-causing mutations were screened in the following genes:GCK-HNF1A-HNF4A-HNF1B-PDX1-NEUROD1-KLF11-CEL-PAX4-INS-BLK. Clinical and laboratory features were compared between children with MODY and children without MODY. RESULTS: The most common reasons for MODY screening were as follows (n/%):low daily dose of insulin (DDI) requirement (122/53%), absence of beta-cell antibodies(58/25.3%), coincidental hyperglycemia(26/11.3%), family history of diabetes (12/5.2%), hypoglycemia/hyperglycemia episodes(7/3%), hyperglycemia related to steroids(3/1.4%) and renal glycosuria(2/0.8%). The markers with the most likelihood to distinguish MODY from T1DM were determined as follows: measurable C-peptide in follow-up, family history of early-onset diabetes and low DDI requirement (odds ratio:12.55, 5.53 and 3.43, respectively). The distribution of the most common causative genes in children with MODY(n = 24) is as follows (n/%):GCK(15/62.5%), HNF4A(7/29.1%), HNF1A(1/9.2%) and PDX1(1/9.2%).All children(n = 12) with GCK-MODY(MODY2) were screened for low DDI requirement, while beta-cell negativity was more common in HNF4A-MODY(MODY1). CONCLUSION: The study shows that measurable C-peptide in follow-up, family history of early-onset diabetes, and low DDI are still remarkable clues to predict MODY in children with misdiagnosed T1DM. In addition, the most common mutations were found in the GCK and HNF4A genes. Among children misdiagnosed with T1DM, a low DDI requirement was found more frequently in MODY2, whereas beta-cell antibody negativity was more common in MODY1.

Increased Expressions of ICAM-2 and ICAM-3 in Pterygium.

Purpose: Pterygium, one of the most common ocular surface diseases, is characterized by inflammatory infiltrates, proliferation, angiogenesis, fibrosis, and extracellular matrix breakdown. The objective of this study was to elucidate the levels of the intercellular adhesion molecule (ICAM)-2, and ICAM-3 gene and protein expressions in pterygium. Methods: A total of 59 patients with pterygium were included in this study. mRNA from pterygial and conjunctival autograft tissues were extracted, and real-time polymerase chain reaction on the BioMark HD dynamic array system was performed for the ICAM-2 and ICAM-3 gene expressions. ICAM-2 and ICAM-3 protein expressions using western blot and immunohistochemistry methods were also investigated in pterygial and conjunctival autograft tissues. Results: ICAM-2 and ICAM-3 gene expressions were markedly augmented in pterygial tissues (P = 0.0018 and P = 0.0023, respectively). Significant increases in protein expressions in pterygial tissues were also detected for ICAM-2 and ICAM-3 (P = 0.0116 and P = 0.0252, respectively). In the immunohistochemical studies, there was a marked increase in ICAM-3 (P = 0.0152), but not in ICAM-2 (P = 0.1041), protein expressions in pterygial tissues. Significant positive correlations between pterygia grading with ICAM-2 protein expression (P = 0.0398) and ICAM-3 immunohistochemical scores (P = 0.0138) were observed. Conclusion: These results demonstrate, for the first time, the expressions of ICAM-2 and ICAM-3 in the pterygium. These findings may help to understand the signal transduction mechanisms in the pterygium formation and provide a new therapy strategy for pterygium treatment.

Chronotypical characteristics and related miR-142-3p levels of children with attention deficit and hyperactivity disorder.

To compare children with Attention Deficit and Hyperactivity Disorder (ADHD) and a healthy control group in terms of chronotype characteristics and miRNA-142-3p/miRNA-378 levels. 50 children with ADHD and 44 healthy children were included in the study. Childhood Chronotype Questionnaire was used to identify the chronotype preferences of children. Serum miR-142-3p and miR- 378 levels were determined. Preference for nighttime was higher in children with ADHD. Additionally, a night preference was found to be associated with attention deficit in both groups. While a significant correlation was found between the psychopathology rate in mothers and the presence of ADHD, there was no such correlation in fathers. In the comparison between children with ADHD and the control group, no significant difference was found between miRNA levels. Both the miR-142-3p and miR-378 values of the children with ADHD that have immediate relatives with a psychiatric disorder were lower, compared to control group. We found that shift to night preference in the circadian rhythm was higher and this preference was associated with attention deficit in the children with ADHD. In addition, the presence of psychopathology in the family and the mother's psychopathology affected the miR-142-3p and miR378 levels.

Does the serotonin receptor gene (rs6313 and rs6314) polymorphism have a role in suicidal attempts?

Suicide is an important public health problem. The aim of the present study is to determine the incidence of serotonin receptor gene polymorphisms (rs6313and rs6314) in patients with a history of suicide attempt by blood sampling and to evaluate whether a causal relation exists between gene polymorphisms and suicide. After obtaining the necessary approvals for the study, we included 178 patients with attempted suicide history admitted to the emergency room between December 14, 2016 and July 31, 2016; 174 control subjects were also included. The blood samples were tested for rs6313 and rs6314 polymorphisms. Among the 178 cases with attempted suicide history, 116 (65.2%) were females and 62 (35.8%) were males. With regard to rs6313 polymorphisms in the case group, 40 cases had AA genotype, 99 had AG genotype, and 39 had GG genotype. In the control group, 38 subjects had AA genotype, 91 had AG genotype, and 45 had GG genotype. With regard to rs6314 polymorphisms, 176 cases in the case group had AG genotype and two cases had GG genotype in the case group, whereas 171 subjects in the control group had AG genotype and three subjects had GG genotype in the control group. The present study did not find any significant association between the incidence of rs6313 and rs6314 polymorphisms and suicidal behavior.

The investigation of miR-221-3p and PAK1 gene expressions in breast cancer cell lines.

The most common malignancy in women is breast cancer. Drug resistance in the treatment of cancer still remains a major clinical concern. Resistance to tamoxifen is seen in half of the recurrences in breast cancer. The anti-estrogen tamoxifen gains agonistic property by transactivating ERα. PAK1-mediated phosphorylation of serine 305 (S305) of ERα leads to resistance to tamoxifen. In our study, PAK1-induced suggestive tamoxifen resistance was designed. According to our hypothesis, phosphorylation of ERα-S305 by PAK1 may be reversed by PAK1 transcriptional inhibition by miR-221-3p due to miR-221-3p targeting the 3' UTR of PAK1. For this purpose, we used Real-time PCR (qRT-PCR) to measure the expression level of miR-221-3p in ER-positive breast cancer cell lines (ZR-75-1, MCF7) and breast epithelial cell line, hTERT-HME1, as control in the laboratory in our department. The increase in the expression of PAK1 depending on miR-221-3p may be related to ZR-75-1 cell line which has invasive characteristic but other two ER+ cancer cell lines, MCF7 and HCC1500, have milder cancer severity. miR-221-3p may have a role on regulation of PAK1 expression because miR-221-3p expression level decreases while PAK1 expression level increases in SKBR3 cell line. miR-221-3p and PAK1 expressions in MDA-MB-231 cell line are higher than that of hTERT-HME1 cell line. This may mean that miR-221-3p has no regulatory effect on of PAK1 expression in this cell line. According to these results, miR-221-3p may give crucial information about molecular mechanism of the disease upon PAK1 activity or different mechanisms with respect to histopathology and severity of breast cancer.

Evaluation of Factor V G1691A, prothrombin G20210A, Factor XIII V34L, MTHFR A1298C, MTHFR C677T and PAI-1 4G/5G genotype frequencies of patients subjected to cardiovascular disease (CVD) panel in south-east region of Turkey.

Cardiovascular disease (CVD) risk factors, such as arterial hypertension, obesity, dyslipidemia or diabetes mellitus, as well as CVDs, including myocardial infarction, coronary artery disease or stroke, are the most prevalent diseases and account for the major causes of death worldwide. In the present study, 4,709 unrelated patients subjected to CVD panel in south-east part of Turkey between the years 2010 and 2013 were enrolled and DNA was isolated from the blood samples of these patients. Mutation analyses were conducted using the real-time polymerase chain reaction method to screen six common mutations (Factor V G1691A, PT G20210A, Factor XIII V34L, MTHFR A1298C and C677T and PAI-1 -675 4G/5G) found in CVD panel. The prevalence of these mutations were 0.57, 0.25, 2.61, 13.78, 9.34 and 24.27 % in homozygous form, respectively. Similarly, the mutation percent of them in heterozygous form were 7.43, 3.44, 24.91, 44.94, 41.09 and 45.66%, respectively. No mutation was detected in 92 (1.95%) patients in total. Because of the fact that this is the first study to screen six common mutations in CVD panel in south-east region of Turkey, it has a considerable value on the diagnosis and treatment of these diseases. Upon the results of the present and previous studied a careful examination for these genetic variants should be carried out in thrombophilia screening programs, particularly in Turkish population.

Screening of common and novel familial mediterranean fever mutations in south-east part of Turkey.

Familial mediterranean fever (FMF) is an autosomal recessive autoinflammatory disorder (MIM# 249100), particularly common in populations of Mediterranean extraction. MEFV gene, responsible for FMF, encoding pyrin has recently been mapped to chromosome 16p13.3. In the present study, 3,341 unrelated patients with the suspicion of FMF in south-east part of Turkey between the years 2009 and 2013 were enrolled and genomic sequences of exon 2 and exon 10 of the MEFV gene were scanned for mutations by direct sequencing. We identified 43 different type of mutations and 9 of them were novel. DNA was amplified by PCR and subjected to direct sequencing for the detection of MEFV gene mutations. Among the 3,341 patients, 1,598 (47.8 %) were males and 1,743 (52.1 %) were females. The mutations were heterozygous in 806 (62.3 %), compound heterozygous in 188 (14.5 %), homozygous in 281 (21.8 %) and mutations had complex genotype in 17 (1.32 %) patients. No mutation was detected in 2,051 (61.4 %) patients. The most frequent mutations were M694V, E148Q, M680I(G/C) and V726A. We could not find any significant differences between the two common mutations according to the gender. Molecular diagnosis of MEFV is a useful tool in clinical practice, thus a future study relating to genotype/phenotype correlation of FMF in more and larger group in Turkish population involving the whole MEFV gene mutations is necessary.