Investigation of effects of olea europaea leaf, rubus fruticosus and nigella CV extracts on microrna expression levels on colon cancer cell lines

Kolorektal kanser, dünyada en yaygın görülen kanserlerden biridir. Son 30 yılda gelişen cerrahi ve görüntüleme tekniklerine rağmen kemoterapatiklerin tedavide yeterince etkin sonuçlar veremediği görülmektedir. Biyoaktif ajanlar olarak adlandırılan bitki özütleri, son yıllarda yeni tedavilerin geliştirilmesi açısından yoğun olarak araştırılmaktadır. Epigenetik mekanizmalar üzerinden etkili olduğu anlaşılan bitkisel özütlerin yeni kanser tedavi modellerinin geliştirilmesinde faydalı olacağı düşünülmektedir. Tüm bu sebepler ile mevcut tez çalışmasında kolorektal kanser hücrelerinde bitkisel özütlerin etkilerinin, önemli bir epigenetik mekanizma olan mikroRNA (miRNA) ekspresyon seviyelerindeki değişimler ile değerlendirilmesi amaçlanmıştır. Gerçekleştirilen çalışmada, Nigella sativa (çörekotu) tohum özütü (NTE), Olea europeae (zeytin) yaprağı özütü (OLE) ve Rubus fruticosus (böğürtlen) kök özütü (RKE)'nün iki kolorektal kanser hücre hattındaki (HT-29 ve LoVo) anti kanser etkisi hücre proliferasyonu ve invazyonu açısından araştırıldı ve olası anti kanser etkinin moleküler mekanizması miRNA ekspresyon seviyesinde çalışılarak epigenetik olarak değerlendirildi. WST-1 analizi sonuçlarına göre değerlendirilen bitki özütlerinin hücre proliferasyonunu anlamlı düzeyde azaltıcı etkisinin (p<0.05) olduğu belirlendi. RKE ve NTE'nin her iki hücre hattında da istatistiki olarak anlamlı seviyede (p<0.05), OLE'nin ise LoVo hücre hattında istatistiki anlamlılığa yakın seviyede (p=0.0641) hücrelerin migrasyon özelliğini inhibe ettiği görüldü. Bitki özütlerinin RT-PCR yöntemi ile miRNA ekspresyon profillerine etkisi değerlendirildiğinde RKE'nin her iki hücre hattında da miR-140 ekspresyon artışına (p<0.05), NTE ve OLE'nin ise HT-29 hücrelerinde miR-182a ekspresyonunda istatistiki olarak anlamlı seviyede (p<0.05) düşüşe neden olduğu belirlendi. Mevcut çalışmada, RKE, NTE ve OLE'nin kolorektal kanser hücrelerinde epigenetik mekanizmalar aracılığı ile hücre proliferasyonu ve migrasyonunu azaltıcı yönde etkiye sahip olduğu literatürde ilk kez ortaya koyulmuştur. İleri çalışmalar gerekmekle birlikte her üç bitki özütünün de kolorektal kanserde yeni tedavilerin araştırılmasına yönelik çalışmalar için potansiyel birer aday olabileceğini düşünmekteyizColorectal cancer one of the most common cancers in the World. Although surgical and screening techniques have been improved a lot in last 30 years, chemotherapeutics have not advanced sufficiently for a successful treatment. New treatments options for many disease by the help of plant extracts, which are called bioactive agents, have been searching intensively. Plant extracts, also effective on epigenetic mechanisms, are thought to be efficious for developing new cancer therapy approaches. Aim of this dissertation study is to evaluate changes in one of the important epigenetic mechanisms microRNA (miRNA) expression levels due to effects of plant extracts on the colorectal cell lines. In present study, Nigella sativa (black cumin) seed extract (NTE), Olea europeae (olive) leaf extract (OLE) and Rubus fruticosus (blackberry) root extract (RKE) are investigated in two colorectal cell lines (HT-29 and LoVo) for their anti-proliferative and migration properties in means of effects on microRNA expressions. Anti-proliferative effects of plant extract are evaluated by WST-1 assay and cell proliferation is reduced significantly (p<0.05). In cell migration analysis RKE and NTE reduced migration in both cell line significantly (p<0.05). Migration properties of LoVo cell line is found to be inhibited by OLE with nearly significant values (p<0.06). Real time-PCR method is used for investigation of expression differences in miRNA after extract treatment. Statistically significant increases are observed in miR-140 expressions in both cell lines after RKE treatment, and in HT-29 cell line miR-182a expression is again significantly reduced after both OLE and NTE treatments (p<0.05). Our data show that NTE, OLE, RKE have capability to inhibit colorectal cancer cell proliferation and migration by the means of epigenetic mechanisms for the first time in the literature. Although further studies should be conducted in this topic, all three plant extracts are thought to be potential candidates for future researches on new therapy options for colorectal cancer

Evaluation of SLC6A2 and CYP2D6 Polymorphisms’ Effects on Atomoxetine Treatment in Attention Deficit and Hyperactivity Disorder

Background There is insufficient replicated data to establish a relationship between the polymorphisms of SLC6A2 and CYP2D6 and the treatment responses of atomoxetine (ATX) in ADHD. We focused on evaluating the effect of top-line single nucleotide polymorphisms (SNPs) in SLC6A2 and CYP2D6 on the ATX treatment response in attention deficit and hyperactivity disorder (ADHD). Methods Of 160 patient records, 34 patients who met the inclusion criteria were evaluated to determine the relationship between genotypes of ten SNPs (six of SLC6A2 and four of CYP2D6) and ATX treatment response. Additionally, the connection between SNPs of CYP2D6 and the severity of side effects associated with ATX was analyzed in 37 patients, including the 34 study patients, and three patients discontinued because of ATX-dependent side effects. Results All six polymorphisms we studied in SLC6A2 were associated with the treatment response of ATX. Clinical improvement in oppositional defiant disorder symptoms of patients with ADHD was only observed in carriers of the homozygous “C” allele of rs3785143 (podd = 0.026). We detected an association between higher CGI-side-effect severity scores and the “TT” genotype of rs1065852 polymorphism in CYP2D6 (p = 0.043). Conclusions The findings of this study suggest that genotypes of polymorphisms within the SLC6A2 and CYP2D6 may play an influential role in treatment response or the severity of side effects associated with ATX in ADHD patients

Contribution of genotypes in Prothrombin and Factor V Leiden to COVID-19 and disease severity in patients at high risk for hereditary thrombophilia

Thrombotic and microangiopathic effects have been reported in COVID-19 patients. This study examined the contribution of the hereditary thrombophilia factors Prothrombin (FII) and Factor V Leiden (FVL) genotypes to the severity of COVID-19 disease and the development of thrombosis. This study investigated FII and FVL alleles in a cohort of 9508 patients (2606 male and 6902 female) with thrombophilia. It was observed that 930 of these patients had been infected by SARS-CoV-2 causing COVID-19. The demographic characteristics of the patients and their COVID-19 medical history were recorded. Detailed clinical manifestations were analyzed in a group of cases (n = 4092). This subgroup was age and gender-matched. FII and FVL frequency data of healthy populations without thrombophilia risk were obtained from Bursa Uludag University Medical Genetic Department's Exome Databank. The ratio of males (31.08%; 27.01%) and the mean age (36.85 & PLUSMN; 15.20; 33.89 & PLUSMN; 14.14) were higher among COVID-19 patients compared to non-COVID-19 patients. The prevalence of FVL and computerized tomography (CT) positivity in COVID-19 patients was statistically significant in the thrombotic subgroup (p < 0.05). FVL prevalence, CT positivity rate, history of thrombosis, and pulmonary thromboembolism complication were found to be higher in deceased COVID-19 patients (p < 0.05). Disease severity was mainly affected by FVL and not related to genotypes at the Prothrombin mutations. Overall, disease severity and development of thrombosis in COVID-19 are mainly affected by the variation within the FVL gene. Possible FVL mutation should be investigated in COVID-19 patients and appropriate treatment should be started earlier in FVL-positive patients

Batı karadeniz bölgesinde obezite prevelansı: Melen çalışması

Purpose: The aim of our study was to determine the prevalence of obesity and abdominal obesity in the West Black Sea region of Turkey and to display the associated chronic diseases, especially Diabetes Mellitus (DM) in this population. Material and Method: We evaluated 2222 (1418 female, 804 male, mean age: 50) participants in Yigilca. The medical histories were recorded and physical examinations were done in all participants. Body weight categories were defined according to the body mass index (BMI) as follows: BMI 94 cm in males and >90 cm in females were accepted as the cut-off points for abdominal obesity. Results: The mean BMI of the participants was 30.6 in females and 27.5 in males. According to BMI, obesity prevalence was 53.1% in females and 26.9% in males. The mean obesity prevalence was 43.5% in general. Abdominal obesity prevalence was 63% in females, 46% in males and 57% in all participants. Obesity prevalence increased with age in both sexes. Especially 3/4 (75%) of females and 1/3 (33%) of males aged 50-59 years were obese. Postmenopausal females had very high (64%) obesity prevalence but the rate was %43 in premenopausal women. While DM prevalence was 12.6% according to patient history, DM prevalence increased to 18.8% by adding patients with fasting glucose >126 mg/dl. Discussion: Obesity, especially abdominal obesity and DM prevalence was found to be at exaggerated rates in both sexes being highest in postmenopausal females

Cystatin C Levels in Patients With Dipper and Nondipper Hypertension

WOS: 000301976100011PubMed: 22373662Objective: Subjects with nondipper hypertension carry a higher risk of cardiovascular events than their normotensive counterparts. The present study was designed to investigate cystatin C levels in patients with dipper and nondipper hypertension. Methods: Eighty-eight consecutive patients who had been treated with antihypertensive drugs for at least 6 months were included in the study. Dipping and nondipping patterns were detected with ambulatory blood pressure monitoring. Clinical, laboratory, and ambulatory blood pressure monitoring data of patient groups with nondipper and dipper hypertension were compared. Results: Patients in the nondipper group were older than those in the dipper group. Serum cystatin C level was higher in the patients in the nondipper group. Cystatin C was negatively correlated with the rate of systolic blood pressure fall at night (r = -0.41; P < 0.001). Linear regression analyses revealed that only cystatin C level was a significant correlate of nocturnal systolic blood pressure decrease. Logistic regression analyses also showed that cystatin C was an independent predictor of nondipping pattern (odds ratio, 3.586; 95% confidence interval, 1.432-8.98; P = 0.006]). Conclusion: The present study showed that cystatin C is higher in patients with nondipper hypertension patients

Dpyd C.19051G&gt;A Promotes Fluoropyrimidine-Induced Anemia, A Prognostic Factor In Disease-Free Survival, In Colorectal Cancer

Background and Aim: In 10-30 of colorectal cancer (CRC) patients, toxic reactions occur after fluoropyrimidine-based chemotherapy. A dihydropyridine dehydrogenase (DPYD) gene variant, c.1905 1G&gt;A, leads to intolerance to fluoropyrimidines. Due to the low frequency of this variant in many populations, the prevalence of fluoropyrimidine-induced hematologic side effects in CRC patients with the c.1905 1G&gt;A variant is unclear. In this study, we investigated the prevalence of the DPYD c.1905 1 variants in a Turkish CRC cohort and the potential effects of these variants on fluoropyrimidine-induced hematologic side effects. Materials and Methods: The DPYD c.1905 1 variant was genotyped using polymerase chain reaction-restriction fragment length polymorphism analysis and confirmed by Sanger sequencing in peripheral blood samples of 100 CRC patients who received fluoropyrimidine-based chemotherapy and 60 healthy volunteers. The association of c.1905 1 variants with susceptibility to hematologic side effects was evaluated. Results: The DPYD c.1905 1G&gt;A variant was more common in the CRC group than in the healthy control group (p 0.001). The presence of the c.1905 1G&gt;A variant was associated with thrombocytopenia (p 0.039) and anemia (p 0.035). CRC patients with fluoropyrimidine-induced anemia had shorter disease-free survival than CRC patients without fluoropyrimidine-induced anemia (p 0.0009). Conclusions: Before administering fluoropyrimidine-based chemotherapy, genetic screening for the DPYD c.1905 1G&gt;A variant should be performed with the aim of preventing anemia and anemia-induced complications in CRC patients.GENETIC TESTING AND MOLECULAR BIOMARKER

Asymmetric dimethylarginine levels in dipper and nondipper hypertensive patients

Amaç: Asimetrik dimetil arjinin (ADMA), nitrik oksit sentezinin endojen bir inhibitörü olup aynı zamanda endotel disfonksiyonunun bir göstergesidir. Non-dipper hipertansiyon (HT) tipine sahip hastalarda yapılan çalışmalarda endotel disfonksiyonun dolayısıyla hedef organ hasarının dipper HT tipi olan hastalardan daha fazla olduğu gösterilmiştir. Biz çalışmamızda bu iki grup arasında endotel fonksiyonu değişikliklerini ADMA seviyelerine bakarak karşılaştırdık. Gereç ve yöntem: Bu çalışmaya 6 ay içerisinde Düzce Üniversitesi Tıp Fakültesi iç hastalıkları ve kardiyoloji polikliniğine başvuran ve daha önceden esansiyel HT tanısı konulup medikal tedavi ile takip edilen 87 hasta dahil edildi. Hastalar ambulatuar kan basıncı ölçümü yapılarak dipper ve non-dipper olmak üzere iki gruba ayrıldı. Hastaların vucut kitle indeksi(VKİ), sistolik kan basıncı ve diyastolik kan basıncı, trigliserid, total kolesterol, düşük dansiteli lipoprotein(LDL) kolesterol, yüksek dansiteli lipoprotein(HDL) kolesterol ve ADMA ölçümleri yapıldı. Bulgular: Grupların yaş, VKİ ve lipid değerleri arasında istatistiksel olarak fark yoktu (p0.05). Dipper grubunda bakılan ADMA seviyeleri 1.290.17 ?mol/L, non-dipper grubunda ise 1.270.13 ?mol/L idi. Dipper ve non-dipper grupları arasında ADMA açısından anlamlı farlılık yoktu (p0.575). Sonuç: ADMA seviyeleri HT hastalarında bozulmuş endotel fonksiyonuna bağlı olarak yüksek olarak bulunur. Kan basıncının non-dipper tipinde endotel disfonksiyonu dipper tipi olanlara göre daha fazladır. Çalışmamızda iki tip arasında ADMA seviyelerinde farklılık bulunamamıştır.Objectives: Asymmetric Dimethylarginine (ADMA) is an endogenous inhibitor of nitric oxide synthesis and also is an indicator of endothelial dysfunction. Patients who have nondipper blood pressure pattern have been shown to exhibit more endothelial dysfunction than patients who has dipper blood pressure pattern. However, other endogenous markers, but not ADMA, are used to assess endothelial function in these studies. In our study, we used ADMA levels to compare differences in endothelial function between each groups. Materials and methods: This study includes 87 patients who admitted to Düzce University internal medicine and Cardiology outpatient clinics throughout 6 months and have been diagnosed previously as essential hypertension and followed with medical therapy. Patients were divided into two groups as non-dippers and dippers, using ambulatory blood pressure measurement. Patients&#8217; body mass index, Systolic blood pressure and diastolic blood pressure, triglycerides, total cholesterol, low density lipoprotein (LDL) cholesterol, high density lipoprotein (HDL) cholesterol, and ADMA levels were measured. Results: Age, BMI, and lipid values did not differ significantly between groups (p>0.05). Mean ADMA level was 1.29±0.13 in dippers, and that of non-dippers was 1.27±0.13&#956;mol/L. ADMA levels did not differ significantly between the non-dipper and dipper groups (p0.575). Conclusion: ADMA levels were found to be higher in hypertensive patients due to endothelial dysfunction. Endothelial dysfunction was more frequent in patients who had nondipper blood pressure pattern than patients who had dipper blood pressure pattern. In our study, ADMA levels did not differ significantly between two patterns

Cystatin C Levels in Patients with Dipper and Nondipper Hypertension

Objective Subjects with nondipper hypertension carry a higher risk of cardiovascular events than their normotensive counterparts. The present study was designed to investigate cystatin C levels in patients with dipper and nondipper hypertension. Methods Eighty-eight consecutive patients who had been treated with antihypertensive drugs for at least 6 months were included in the study. Dipping and nondipping patterns were detected with ambulatory blood pressure monitoring. Clinical, laboratory, and ambulatory blood pressure monitoring data of patient groups with nondipper and dipper hypertension were compared. Results Patients in the nondipper group were older than those in the dipper group. Serum cystatin C level was higher in the patients in the nondipper group. Cystatin C was negatively correlated with the rate of systolic blood pressure fall at night ( r = −0.41; P &lt; 0.001). Linear regression analyses revealed that only cystatin C level was a significant correlate of nocturnal systolic blood pressure decrease. Logistic regression analyses also showed that cystatin C was an independent predictor of nondipping pattern (odds ratio, 3.586; 95% confidence interval, 1.432–8.98; P = 0.006]). Conclusion The present study showed that cystatin C is higher in patients with nondipper hypertension patients. </jats:sec