A DNA Extraction Method for Nondestructive Testing and Evaluation of Cotton Seeds (Gossypium L.).

Kernels of cotton provide lint and linter for textiles, oil and protein for food and feed. Cotton seed is formed following fertilization between an ovule and a pollen grain. The seed coat is maternal in origin, whereas the embryo and attached cotyledonary leaves are hybrids of parental lines. The extraction of genomic DNA from an ungerminated whole, a portion or mixed seeds are prerequisite in genetic and genomic studies of cotton. As far as our knowledge, there is only one method of nondescriptive DNA extraction from ungerminated cotton seeds without affecting the seed germination capability, but it has technical difficulties and requires special equipment. Furthermore, the amount of DNA extracted using the published method is low and, therefore, it is only suitable for routine marker assisted selection studies. In this study, a DNA extraction protocol referred to as the CTAB-LiCl was developed for single whole cotton seed, a portion of cotton seed and bulked cotton seeds. This protocol uses a combination of CTAB and LiCl to lyse cells and deplete RNAs simultaneously. The CTAB-LiCl DNA extraction method was evaluated in ninety-six individuals of six different cotton cultivars along with two genetic standards of cotton, TM-1 (G. hirsutum L.), Pima 3-79 (G. barbadense L.), and several other plant species of different plant genera. Results revealed that this method produced high quality and amounts of DNA as confirmed by spectrophotometry, agarose gel, restriction enzyme digestion, polymerase chain reaction, and library production for next generation sequencing studies of whole genome bisulfite sequencing. It does not require the use of liquid nitrogen, RNase, proteinase K, or beta-mercaptoethanol and can be completed in approximately 2 h. Small tissues of the chalaza ends of ungerminated cotton seeds could be used to obtain high quality and quantity of DNA ranging from 14 to 28 µg without affecting the seeds' germination ability, allowing marker-assisted selection before planting and flowering.

Grafting based DNA methylation alteration of snoRNAs in upland cotton (Gossypium L.).

The effects of grafting in response to various biotic and abiotic stressors have been studied, however, the methylation status of small nucleolar RNA (snoRNA) genes in heterograft and homograft cotton needs investigation. This study was undertaken to determine grafting effects on DNA methylation of snoRNA genes in Upland cotton. Rootstocks used were Pima 3-79 (Gossypium barbadense acc. Pima 3-79) and Texas Marker-1 (G. hirsutum acc. TM-1), representing two different species with different fiber properties, adaptations, and morphologies. The methylation ratio and differently methylated cytosines (DMCs) of 10935 snoRNA genes in mature seeds of heterograft and homograft cotton samples were studied using the whole genome bisulfite sequencing method. Seedling vigor and seed weight were studied to investigate phenotype alterations that might be associated with altered methylation levels among grafts. Statistically significant DMC differences among gene elements of snoRNA genes and between homograft and heterograft cotton samples were identified in the absence of DNA sequence alterations. DNA methylation alterations of snoRNA genes associated with seedling vigor and 100 seed weight. The majority of snoRNA genes showed higher numbers of mCG + mCHG-DMCs with increased methylation levels in heterograft, while there were higher numbers of mCG + mCHG-DMCs with decreased methylation levels in homograft. Since snoRNAs regulate essential genes for plant growth and development and plant adaptation to different habitats or extreme environments, their altered methylation levels should be related with plant physiology.

Comparison of ICSI and conventional IVF in non-male factor patients with less than four oocytes.

BACKGROUND: We aimed to analyse our clinical results for a particular subgroup of patients with poor ovarian response (POR) to clarify if lower number of oocytes is a drawback for proceeding to C-IVF. MATERIALS AND METHODS: In this retrospective study, patient files of all couples (#1733) who underwent oocyte retrieval between January 2017 and December 2019 were reviewed and 191 cases diagnosed with non-male factor infertility in which ≤ 3 cumulus-oocyte complexes available for fertilisation were analysed. Exclusion criteria were: woman age > 42, patients with a history of previous ART trial, prenatal genetic testing cycles and couples undergoing total cryopreservation for any indication. Three groups were constructed depending on the method of fertilisation and on semen quality as follows: IVF non-male factor (Group 1, n = 77); ICSI non-male factor (Group 2, n = 65); ICSI male factor-ICSI/MF n = 49 according to WHO reference values. Main outcome parameters were: fertilisation rate, implantation rate and live birth rate. RESULTS: Fertilisation rate per collected COC was significantly higher in group 1 compared to the other two groups (85.68%, 72.58%, 73.33% respectively, p = 0.004). FR per inseminated oocyte also tended to be higher in group 1 but not reaching a statistically significant level. Both techniques yielded similar implantation rates (20.42%, 28.49%, 23.33% respectively, p = 0.407) and live birth rates (26.8%, 30.6%, 31.1%, respectively, p = 0.643). CONCLUSION: In the presence of normal semen parameters, low egg number is not an indication to perform ICSI. The choice of fertilisation method should be based primarily on semen quality, in combination with the patient's previous history regardless of the ovarian reserve.

Tissue and/or developmental stage specific methylation of nrDNA in Capsicum annuum.

The nuclear ribosomal DNA (nrDNA) sequences are often used for phylogenetic analysis among organisms. Because DNA cytosine methylation and nucleolar dominancy are two common epigenetic mechanisms of nrDNA, we hypothesized that internal transcribed spacer 1 (ITS1), 5.8S rRNA and ITS2 of nrDNA sequences could be used as epigenetic biomarkers. Thus, this research was undertaken to study level and pattern of site-specific cytosine methylation of ITS1, 5.8S and ITS2 in nine tissues and/or developmental stage of pepper Capsicum annuum L. cultivar Demre Sivrisi. Tissues studied consisted of young and old roots at 30 and 90 days after sowing (das), mature dry seeds and seeds at 26 days of post anthesis (dpa), flowering buds at 1 day before flowering, pericarps at 3, 15 and 65 dpa. Levels and patterns of DNA cytosine methylation were identified at single base resolution using bisulfite conversion sequencing. Results of this study revealed that DNA cytosine level and pattern of ITS1, 5.8S and ITS2 were different in most tissues and/or developmental stages studied. In addition, methylation levels of CG, CHG and CHH contexts were also significantly different among the regions. Based on the findings of this study, it was concluded that high level of methylation of nrDNA sequences was relatively higher as observed in transposable element and promoter. On the other hand, its tissue-specific gene expression was effective as that of gene body and promoter methylation. Overall findings revealed that methylation levels of nrDNA could be used as biomarkers for tissue identification or age estimation in plants.

Molecular characterization of the Pratylenchus vulnus populations on cereals in Turkey.

Pratylenchus vulnus (walnut root lesion nematode) is one of the most damaging root lesion nematode species worldwide. In this study, 17 populations of P. vulnus obtained from wheat and barley cultivated fields in 2016 to 2017 in Turkey (Karaman and Konya provinces) were identified using real-time PCR and melting curve analysis. Samples provided a single peak at 87.3˚C with real-time PCR. D2 to D3 expansion segments of the 28S rRNA of one population from Cihanbeyli district in Konya province was sequenced and recorded in GenBank (Accession number: MT320536.1). Alignments of the population was identical 98.66% to the populations of P. vulnus available in GenBank (Accs. nos: LT985479.1 and LT965052.1) and 98.65% (Accs. nos: KY424305.1 and KY424304.1).

Genotypic-phenotypic features and enzyme replacement therapy outcome in patients with mucopolysaccharidosis VI from Turkey.

Mucopolysaccharidosis type VI (MPS VI) is a lysosomal storage disorder (LSD) characterized by a chronic, progressive course with multiorgan involvement. In our study, clinical, biochemical, molecular findings, and response to enzyme replacement therapy (ERT) for at least 6 months were evaluated in 20 patients with MPS VI. Treatment effects on clinical findings such as liver and spleen sizes, cardiac and respiratory parameters, visual and auditory changes, joints' range of motions, endurance tests and changes in urinary glycosaminoglycan excretions, before and after ERT were analyzed. ERT caused increased physical endurance and decreased urinary dermatan sulfate/chondroitin sulfate ratios. Changes in growth parameters, cardiac, respiratory, visual, auditory findings, and joint mobility were not significant. All patients and parents reported out an increased quality of life, which were not correlated with clinical results. The most prevalent mutation was p.L321P, accounting for 58.8% of the mutant alleles and two novel mutations (p.G79E and p.E390 K) were found. ERT was a safe but expensive treatment for MPS VI, with mild benefits in severely affected patients. Early treatment with ERT is mandatory before many organs and systems are involved.

[A case with scalp pruritus caused by Dermanysus gallinae (order: mesostigmata)]. / Dermanysus gallinae'nin (Takim: mesostigmata) neden oldugu saçli deride kasintili olgu.

Dermanyssus gallinae is one of the important hematophagous ectoparasite species of poultry like chicken, pigeon and wild bird species. These ectoparasites in the form of nymphs or adults who can not find their hosts are also seen in mammals and even in humans. For this reason, they are considered as important for public health. The ectoparasite causes a clinical condition named gamasoidosis among pet owners and people who live or work close to animal shelters, barns and chicken farms. Pruritus dermatitis is also caused by D.gallinae in humans and can cause false diagnosis. In this report, a case of D.gallinae which leads to severe itch in the hairy head skin was presented. A 66-year-old female patient admitted to University Hospital with complaints of "bugs in her hair and itching of the skin increasing in the evenings" that have persisted for a month. In the dermatological examination of the patient, it was noted that her hair and scalp were usual. Routine laboratory tests were normal. However, a large number of mites were found in her headscarf that she brought with her to the examination. Later, it was learned that the patient feeds chicken in her garden in the village where she lives. The collected mite samples were were kept in glass test tubes that contained glycerol and alcohol. The mites were identified as D.gallinae by morphological identification with light microscopy by using 10x, 20x and 40x magnifications. The mites were described as D.gallinae (Order: Mesostigmata, local name: poultry red mite, perch mite, poultry mite) with the morphological examination. Long-acting 1% permethrin shampoo was applied to remove the mites on the patient and during the controls, it was changed as 5% permethrin and 10% crotamiton lotion. For environmental sanitation, carbamates (such as carbolineum, trichlorfon, malathion, tetrachlorvinphos, etc.), organophosphates and acaricide insecticides with pyrethroids spraying or powder formulations were recommended. It was recommended to repair the slits and cracks where the parasite in the shelter could be stored. The patient was informed on (i) how to clean the household items with susceptible acaricides, (ii) removal of unused infected animal shelters, cages and nests from human habitat, (iii) raising of ambient temperature above 45°C, (iv) ventilation of the living spaces and (v) washing the clothes with detergent. In order to be effectively protected from the risk of infection and the detriments that are brought by this parasite, it is imperative to stay away from the hosts and the infected areas such as chicken farms, to obey the hygiene regulations, and to properly conduct the disinfestation of the shelters. In addition, it is also helpful to receive a true story from the patient, with details of contact with birds for the protection and treatment.

Monocyte to high-density lipoprotein ratio as a new prognostic marker in patients with STEMI undergoing primary percutaneous coronary intervention.

BACKGROUND: Monocyte count to high-density lipoprotein ratio (MHR) has recently emerged as an indicator of inflammation and oxidative stress in the literature. We aimed to investigate the prognostic value of MHR in patients with ST-segment elevation myocardial infarction treated with primary percutaneous coronary intervention (PCI). METHODS: A total of 513 patients who were hospitalized with diagnosis of acute ST-segment elevation myocardial infarction and treated with primary PCI were retrospectively enrolled in the study. Demographic and clinical data, admission laboratory parameters, and MHR values were recorded. Inhospital major adverse cardiac events (MACE) and mortality were reported as the clinical outcomes. RESULTS: Twenty-six patients (5%) died, and MACE was observed in 86 patients (17%) during hospital follow-up. Patients were categorized in 3 groups according to tertiles of admission MHR. The rates of inhospital mortality and MACE were significantly higher in tertile 3 group compared to tertile 1 group (10% vs 1%, 27% vs 11%; P < .01 and P < .01). In multivariate regression analysis, age, sex, presence of Killip 3 or 4, left ventricular ejection fraction, troponin I, C-reactive protein, and increased MHR levels (odds ratio, 1.03; 95% confidence interval, 1.01-1.05; P < .01) independently predicted inhospital mortality; age, presence of Killip 3 or 4, troponin I, and increased MHR levels (odds ratio, 1.02; 95% confidence interval, 1.01-1.04; P < .01) independently predicted MACE. CONCLUSION: Admission MHR values were found to be independently correlated with inhospital MACE and mortality after primary PCI.

Aliskiren - a promising strategy for ovarian ischemia/reperfusion injury protection in rats via RAAS.

The aim of this study was to evaluate the effects of aliskiren, direct renin inhibitor, as an antioxidant and tissue protective agent and evaluate the molecular, biochemical, and histopathological changes in experimental ischemia and ischemia/reperfusion injury in rat ovaries. Forty-eight female rats were randomly divided into eight groups: in Group 1, only sham operation was performed. Group 2 received 100 mg/kg aliskiren and sham operated. In Group 3, 3 h-period of bilateral ovarian ischemia was applied. Group 4 received a 3-h period of ischemia followed by 3 h of reperfusion. Groups 5 and 6 received 50 and 100 mg/kg, respectively, of aliskiren and bilateral ovarian ischemia was applied (after a 3-h period of ischemia, both ovaries were surgically removed). To Groups 7 and 8, 50 and 100 mg/kg of aliskiren were administered, respectively, and the induction of ischemia was performed. At the end of a 3-h period of ischemia, bilateral vascular clips were removed, and 3 h of reperfusion continued. After the experiments, IL-1ß, IL-6, TNF-α, and iNOS mRNA expressions and SOD, GSH, MDA, renin, and angiotensin-II levels were determined and histopathological changes were examined in rat ovaries. Aliskiren treatment normalized excessive changes in cytokine and oxidative stress markers in both ischemia and ischemia/reperfusion injury. Histopathologically, treatment with aliskiren ameliorated the development of ischemia and/or ischemia/reperfusion tissue injury. This study concluded that aliskiren treatment is effective in reversing ischemia and/or ischemia/reperfusion induced ovary damage via the improvement of oxidative stress, reduction of inflammation, and suppression of the renin-angiotensin aldosterone system.

Genetic diversity of disease-associated loci in Turkish population.

Many consortia and international projects have investigated the human genetic variation of a large number of ethno-geographic groups. However, populations with peculiar genetic features, such as the Turkish population, are still absent in publically available datasets. To explore the genetic predisposition to health-related traits of the Turkish population, we analyzed 34 genes associated with different health-related traits (for example, lipid metabolism, cardio-vascular diseases, hormone metabolism, cellular detoxification, aging and energy metabolism). We observed relevant differences between the Turkish population and populations with non-European ancestries (that is, Africa and East Asia) in some of the investigated genes (that is, AGT, APOE, CYP1B1, GNB3, IL10, IL6, LIPC and PON1). As most complex traits are highly polygenic, we developed polygenic scores associated with different health-related traits to explore the genetic diversity of the Turkish population with respect to other human groups. This approach showed significant differences between the Turkish population and populations with non-European ancestries, as well as between Turkish and Northern European individuals. This last finding is in agreement with the genetic structure of European and Middle East populations, and may also agree with epidemiological evidences about the health disparities of Turkish communities in Northern European countries.

Detection of biotinidase gene mutations in Turkish patients ascertained by newborn and family screening.

UNLABELLED: The incidence of biotinidase deficiency in Turkey is currently one of the highest in the world. To expand upon the information about the biotinidase gene (BTD) variations in Turkish patients, we conducted a mutation screening in a large series (n = 210) of probands with biotinidase deficiency, using denaturing high-performance liquid chromatography and direct DNA sequencing. The putative effects of novel mutations were predicted by computational program. Twenty-six mutations, including six novels (p.C143F, p.T244I, c.1212-1222del11, c.1320delG, p.V457L, p.G480R) were identified. Nine of the patients were symptomatic at the initial clinical assessment with presentations of seizures, encephalopathy, and lactic acidemia. The most common mutation in this group of symptomatic patients was c.98-104 del7ins3. Among the screened patients, 72 have partial and 134 have profound biotinidase deficiency (BD) of which 106 are homozygous for BTD mutations. The common mutations (p.R157H, p.D444H, c.98-104del7ins3, p.T532M) cumulatively accounted for 72.3% of all the mutant alleles in the Turkish population. CONCLUSION: The identification of common mutations and hot spot regions of the BTD gene in Turkish patients is important for mutation screening in the Turkish population and helps to ascertain carriers, may have impact on genetic counseling and implementing prevention programs.

GSTM1, GSTP1, and GSTT1 genetic variability in Turkish and worldwide populations.

OBJECTIVE: Glutathione S-transferase (GST) variants have been widely investigated to better understand their role in several pathologic conditions. To our knowledge, no data about these genetic polymorphisms within the Turkish population are currently available. The aim of this study was to analyze GSTM1 positive/null, GSTT1 positive/null, GSTP1*I105V (rs1695), and GSTP1*A114V (rs1138272) variants in the general Turkish population, to provide information about its genetic diversity, and predisposition to GST-related diseases. METHODS: Genotyping was performed in 500 Turkish individuals using the Sequenom MassARRAY platform. A comparative analysis was executed using the data from the HapMap and Human Genome Diversity Projects (HGDP). Sequence variation was deeply explored using the Phase 1 data of the 1,000 Genomes Project. RESULTS: The variability of GSTM1, GSTT1, and GSTP1 polymorphisms in the Turkish population was similar to that observed in Central Asian, European, and Middle Eastern populations. The high linkage disequilibrium between GSTP1*I105V and GSTP1*A114V in these populations may have a confounding effect on GSTP1 genetic association studies. In analyzing GSTM1, GSTT1, and GSTP1 sequence variation, we observed other common functional variants that may be candidates for associated studies of diseases related to GST genes (e.g., cancer, cardiovascular disease, and allergy). CONCLUSIONS: This study provides novel data about GSTM1 positive/null, GSTT1 positive/null, GSTP1*I105V, and GSTP1*A114V variants in the Turkish population, and other functional variants that may affect GSTM1, GSTT1, and GSTP1 functions among worldwide populations. This information can assist in the design of future genetic association studies investigating oxidative stress-related diseases.

Ultrasonography for the prediction of extension of trophoblastic infiltration into the tubal wall in ampullary pregnancy.

OBJECTIVE: Predictive factors of damage to the Fallopian tube may guide the treatment for patients with tubal pregnancy. The purpose of this study was to evaluate the predictive value of ultrasonographic findings in patients affected by ampullary pregnancy for the determination of the depth of trophoblastic infiltration into the tubal wall on histological examination. MATERIAL AND METHODS: 38 patients with ampullary pregnancy undergoing salpingectomy were enrolled into the study. The patients were divided into two subgroups depending on their transvaginal sonography (TVS) findings; either an ectopic gestational sac containing an embryo with cardiac activity or those with a tubal ring. The ampullary pregnancies were histologically classified according to the depth of infiltration of trophoblastic tissue into the tubal wall as follows: stage I: limited to mucosa; stage II: extension to the muscularis layer; stage III: complete infiltration of the tubal wall with or without rupture of the serosa. The association between findings on TVS and stage of trophoblastic invasion, serum beta-human chorionic gonodatropin (ß-hCG) levels was evaluated. RESULTS: Although there was no significant difference among two groups in terms of histological stage of trophoblastic infiltration (p = 0.257), patients in whom an embryo with cardiac activity had been identified were found to have higher percentage of stage II (47.8%) or stage III (8.7%) invasion. However, there was a significant difference in serum ß-hCG levels on the day of surgery among the two groups (p = 0.028). CONCLUSIONS: Ultrasonographic aspect of ampullary pregnancy is associated with depth of trophoblastic infiltration into the tubal wall and serum ß-hCG levels.

Long term outcomes of surgical revascularization for isolated left main coronary artery stenosis: a single-center surveillance study.

OBJECTIVE: The objective of this study was to retrospectively analyze the clinical course and postoperative long-term survival of patients diagnosed with isolated left main coronary artery (LMCA) stenosis after surgical revascularization. METHODS: A total of 38 patients (27 males, 11 females) who were diagnosed with isolated LMCA stenosis and underwent surgical revascularization were enrolled in the study. Isolated LMCA stenosis was classified into 2 groups: ostial stenosis and nonostial stenosis. Coronary events were defined as death of cardiac origin, the need for a new myocardial revascularization procedure, or the occurrence of myocardial infarction in the course of follow-up. The postoperative assessment period included short- and long-term follow-up. The study endpoint was defined as all-cause mortality. RESULTS: Among the 38 patients who participated in the study, 25 suffered from ostial LMCA stenosis. The early postoperative mortality rate before hospital discharge was 2.6%. Median duration of postoperative long-term follow-up was 73.43 months (range: 0.17-187.23). Median duration of long-term follow-up free from coronary events or percutaneous coronary interventions was 73.43 months. Postoperative 2-year survival rate was 97.4%, and 5-year survival rate was 92.1%. The postoperative survival period and period free of coronary events of patients with isolated ostial LMCA stenosis did not differ significantly from those of patients with nonostial stenosis (p=0.801, p=0.970, respectively). CONCLUSION: Postoperative short- and long-term prognosis of isolated LMCA stenosis appears good in terms of mortality and coronary event symptoms.

Efficacy of oral hydration in the prevention of contrast-induced acute kidney injury in patients undergoing coronary angiography or intervention.

BACKGROUND: Efficacy of intravenous (IV) volume expansion in preventing contrast-induced acute kidney injury (CI-AKI) is well known. However, the role of oral hydration has not been well established. The aim of this work was to evaluate the efficacy of oral hydration in preventing CI-AKI. METHODS: We prospectively randomized 225 patients undergoing coronary angiography and/or percutaneous coronary intervention in either oral hydration or IV hydration groups. Patients who have at least one of the high-risk factors for developing CI-AKI (advanced age, type 2 diabetes mellitus, anemia, hyperuricemia, a history of cardiac failure or systolic dysfunction) were included in the study. All patients had normal renal function or stage 1-2 chronic kidney disease. Patients in the oral hydration group were encouraged to drink unrestricted amounts of fluids freely whereas isotonic saline infusion was performed by the standard protocol in the IV hydration group. RESULTS: CI-AKI occurred in 8/116 patients (6.9%) in the oral hydration group and 8/109 patients (7.3%) in the IV hydration group (p = 0.89). There was also no statistically significant difference between the two groups when different CI-AKI definitions were taken into account. CONCLUSION: Oral hydration is as effective as IV hydration in preventing CI-AKI in patients with normal kidney function or stage 1-2 chronic kidney disease, and who also have at least one of the other high-risk factors for developing CI-AKI.

Use and effectiveness of prothrombin complex concentrates vs fresh frozen plasma in gastrointestinal hemorrhage due to warfarin usage in the ED.

OBJECTIVES: High International Normalized Ratio (INR) level resulting from warfarin use increases the risk of gastrointestinal hemorrhages. We aimed to compare the efficacy of prothrombin complex concentrates (PCC) and fresh frozen plasma (FFP) at lowering the INR level, decreasing active hemorrhages visible by endoscopy, and shortening the length of stay at the emergency department (ED). METHOD: This study is a prospective cohort study of consecutive patents with gastrointestinal hemorrhages that received either PCC or FFP. With strict exclusion criteria, only patients over 18 years of age with a high INR level (>2.1) due to warfarin usage were included. RESULTS: A total of 40 patients (18 female) were included in the study, 20 each in the PCC and FFP groups. For the PCC group, the mean INR levels at the second and sixth hours were lower than those for the FFP group (second hour INR: 1.53 vs 4.50, P<.01, sixth hour INR: 1.52 vs 2.41, P<.01). Seven patients experienced active bleeding (Forrest 1) in the FFP group, whereas no patient experienced active bleeding in the PCC group based on the Forrest classification (35% vs 0%, P<.01), and only 3 patients in the FFP group underwent invasive/surgical treatment (15% vs 0%, P<.01). The ED length of stay was lower for the PCC group (1.62 days vs 3.46 days, P<.01). CONCLUSION: For patients experiencing a gastrointestinal hemorrhage, INR levels were reversed more quickly, there was less active bleeding on endoscopy, and the ED length of stay was lower in the PCC group than in the FFP group.

Identification of novel NRF2-regulated genes by ChIP-Seq: influence on retinoid X receptor alpha.

Cellular oxidative and electrophilic stress triggers a protective response in mammals regulated by NRF2 (nuclear factor (erythroid-derived) 2-like; NFE2L2) binding to deoxyribonucleic acid-regulatory sequences near stress-responsive genes. Studies using Nrf2-deficient mice suggest that hundreds of genes may be regulated by NRF2. To identify human NRF2-regulated genes, we conducted chromatin immunoprecipitation (ChIP)-sequencing experiments in lymphoid cells treated with the dietary isothiocyanate, sulforaphane (SFN) and carried out follow-up biological experiments on candidates. We found 242 high confidence, NRF2-bound genomic regions and 96% of these regions contained NRF2-regulatory sequence motifs. The majority of binding sites were near potential novel members of the NRF2 pathway. Validation of selected candidate genes using parallel ChIP techniques and in NRF2-silenced cell lines indicated that the expression of about two-thirds of the candidates are likely to be directly NRF2-dependent including retinoid X receptor alpha (RXRA). NRF2 regulation of RXRA has implications for response to retinoid treatments and adipogenesis. In mouse, 3T3-L1 cells' SFN treatment affected Rxra expression early in adipogenesis, and knockdown of Nrf2-delayed Rxra expression, both leading to impaired adipogenesis.

Differences in geographical distribution and risk factors for urinary incontinence in Turkey: analysis of 6,473 women.

OBJECTIVE: To assess the prevalence, types and risk factors for urinary incontinence (UI) and to evaluate the impact of incontinence on quality of life by using validated and objective questionnaires in the western and eastern parts of Turkey. METHODS: In this multicenter observational study, 6,473 women from 38 cities in the western and eastern parts of Turkey were included. UI was assessed by ICIQ-SF (International Consultation on Incontinence Questionnaire Short Form) and IIQ-7 (Incontinence Impact Questionnaire). RESULTS: The UI rate was 20.9% (10% for stress, 8.3% overactive bladder and 2.6% for mixed type). In all, stress incontinence was the most common type. The rate of UI in women residing in the west was higher than in women living in the east (p < 0.001). ICIQ scores were comparable in the two groups but women in the west scored higher in each item of the IIQ. Age >40 years (p < 0.001), number of siblings >5 (p < 0.001) and low educational status (p < 0.001) increased the rate of incontinence. In binary logistic regression analysis menopausal status, age >40 years, number of siblings >5, being overweight, region of residence, and educational status were associated with UI. CONCLUSION: The rate of UI in women residing in the western part of Turkey was higher than women living in the east. Residing in a different geographical region (in our case living either in the western or eastern part of Turkey) seemed to be an independent risk factor for UI. Moreover, UI deteriorates quality of life and more attention should be paid to this vulnerable population.

Betulin Stimulates Osteogenic Differentiation of Human Osteoblasts-Loaded Alginate-Gelatin Microbeads.

Osteoporosis, a terminal illness, has emerged as a global public health problem in recent years. The long-term use of bone anabolic drugs to treat osteoporosis causes multi-morbidity in elderly patients. Alternative therapies, such as allogenic and autogenic tissue grafts, face important issues, such as a limited source of allogenic grafts and tissue rejection in autogenic grafts. However, stem cell therapy has been shown to increase bone regeneration and decrease osteoporotic bone formation. Stem cell therapy combined with betulin (BET) supplementation might be adequate for bone remodeling and new bone tissue generation. In this study, the effect of BET on the viability and osteogenic differentiation of hFOB 1.19 cells was investigated. The cells were encapsulated in alginate-gelatin (AlGel) microbeads. In vitro tests were conducted during the 12 d of incubation. While BET showed cytotoxic activity (>1 µM) toward non-encapsulated hFOB 1.19 cells, encapsulated cells retained their functionality for up to 12 days, even at 5 µM BET. Moreover, the expression of osteogenic markers indicates an enhanced osteo-inductive effect of betulin on encapsulated hFOB 1.19, compared to the non-encapsulated cell culture. The 3D micro-environment of the AlGel microcapsules successfully protects the hFOB 1.19 cells against BET cytotoxicity, allowing BET to improve the mineralization and differentiation of osteoblast cells.

Investigation of the effects of in ovo taurine injection on hatching characteristics and stress reduction potential.

BACKGROUND: In ovo application is the process of administering some nutrients or components into the egg. The main purpose of this application is to ensure that some nutrients are provided to chicks with a short incubation period. Few studies were conducted with taurine in fertile eggs; especially, no observation of hatchability and chick quality has been found. In addition, taurine has an anti-stress impact that fights oxidative factors. OBJECTIVE: To assess the hatchability and chick quality after in ovo taurine administration. To determine the stress that may occur as a result of in ovo application and whether taurine has a stress-reducing effect. METHODS: A total of 1200 fertile eggs from a 34-week-old broiler breeder (Ross 308) flock were categorized into 4 groups with 75 eggs per replicate: control (uninjected), taurine group (0.30 mL dissolved taurine in distilled water), sham control (sterile distilled water) and perforation (eggs perforated and then waxed). On day 14 of incubation, an in ovo injection was administered to the albumen. Data on hatching parameters and hepatic HSP70 levels were obtained using relevant formulas and western blotting, respectively. RESULTS: Control chicks exhibited higher hatchability than other groups, with the taurine group showing the lowest hatchability. The HSP70 levels were the highest in the perforation group compared to the control group. An increase of 21.37% in the taurine group and 83.45% in the sham control group was observed compared to the control group. CONCLUSIONS: The findings suggest that in ovo application may induce increased stress, whereas taurine may have positive effects in mitigating the stress caused by in ovo application.