Purification of alpha-galactosidase from pepino (Solanum muricatum) by three-phase partitioning

WOS: 000297444900018Three-phase partitioning (TPP) is a bioseparation technique used for the extraction, concentration and purification of biomolecules. The technique consists of simultaneous addition of a salt (generally ammonium sulfate) to the crude extract followed by the addition of an organic solvent (generally t-butanol). In the present study, alpha-galactosidase was purified from pepino (Solanum muricatum) fruit by TPP. The influence of various process parameters (ammonium sulfate saturation, crude extract to t-butanol ratio and pH) on alpha-galactosidase partitioning is investigated to get highest purity fold and yield. The results showed that, 50% (w/v) ammonium sulfate saturation with 1:1.5 crude extract to t-butanol ratio at pH 5.25 gave 6.2-fold purification with 127% activity recovery of alpha-galactosidase. Characterization and determination of biochemical properties of the partitioned alpha-galactosidase were also aimed. The sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis showed considerable purification and the molecular weight of the enzyme was nearly found to be as 38 kDa. Various parameters (temperature, pH and substrate concentration) affecting to the enzyme activity and stability were studied. The optimum temperature and pH of the partitioned alpha-galactosidase were found to be 50 degrees C and pH 5.5, respectively. The enzyme was very stable at the temperature ranged from 37 to 45 degrees C and also in a pH range of 4.5-7.0. The kinetic constants; K(m) and V(max) were determined to be 0.37 mM and 0.46 U, respectively. The results indicated that, TPP technique is very attractive process for purification of pepino alpha-galactosidase and the characteristic properties of the enzyme partitioned by TPP make the enzyme good candidates for its several industrial applications. (C) 2011 Elsevier B.V. All rights reserved

Role of the norepinephrine transporter polymorphisms in atomoxetine treatment: From response to side effects in children with ADHD

Objective

The Effects of Wortmannin and EGCG and Combined Treatments on MDA-MB-231 Breast Cancer Cell Lines via Inactivation of PI3K Signaling Pathway

Epigallocatechin gallate (EGCG), a major polyphenol in green tea, has been studied as an agent against carcinogenesis and Wortmannin is a microbial steroid and it inhibits phosphatidyl inositol 3-kinase (PI3K) pathway. The aim of this study was to investigate the effects of PI3K inhibitor Wortmannin, EGCG and combined treatments on PI3K pathway on human breast cancer cell line MDA-MB-231 using indirect immunohistochemistry method. MDA-MB-231 breast cancer cells were cultured in RPMI-1640 medium containing 10% FBS, 1% l-glutamine and 1% penicillin/streptomycin. Anti-PI3K, anti-AKT, anti-ERK, anti-NFkB, anti-c-jun and anti-EZH2 primary antibodies were used for indirect immunohistochemistry after 24 h administrations of Wortmannin (2.5 µM), EGCG (100 µM) and combination of them. The mean values of the staining intensities (mild, moderate, strong and very strong) and percentage of positively stained cells were calculated using H-Score. The results of this study showed that the combined treatment of Wortmannin and EGCG is more effective on the decreasing of immunoreactivities of PI3K pathway molecules than single administrations. The combined use of these drugs is thought to be advantageous in enhancing the development and efficacy of existing cancer treatments

Comparison of ultrastructural changes and the anticarcinogenic effects of thymol and carvacrol on ovarian cancer cells: which is more effective?

Cavusoglu, Turker/0000-0001-7100-7080WOS: 000520568500001PubMed: 32183603Ovarian cancer is the seventh most common cancer worldwide in women. Many anticancer drugs are currently used clinically have been isolated from plant species or are based on such substances. Thymol (5-methyl-2-isopropylphenol) and carvacrol are oxygenated aromatic compounds from the monoterpene group. They are the main constituents of thyme essential oil and show antiproliferative, antioxidant, and antiseptic properties. the aim of this study is to compare the antiproliferative and apoptotic effects of thymol and carvacrol on SKOV-3 ovarian cancer cell line. the cancer cells were treated with different concentrations of thymol and carvacrol (100, 200, 400, 600 mu M) at 24 h and 48 h durations. the cell viability was investigated by MTT assay and analysis of apoptosis with annexin V assay was determined. the study show that thymol and carvacrol significantly induced apoptosis in all groups as dose and time-dependent (p < .05). the data in the present study demonstrated that thymol and carvacrol have apoptotic and antiproliferative properties in a concentration-dependent manner toward ovarian cancer cells. SKOV-3 cancer cell line was much more sensitive to the toxic effect of thymol than carvacrol.Mugla Sitki Kocman University Research Projects Coordination OfficeMugla Sitki Kocman University [17/062]This study has been granted by the Mugla Sitki Kocman University Research Projects Coordination Office through Project Grant Number: (17/062). the funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscrip

The Effects of Paclitaxel and Metformin and Combined Treatments on TLR Signaling Pathway on MDA-MB-231 Breast Cancer Cell Lines

The aim of this study was to determine the effects of Paclitaxel, Metformin and combined treatments on human breast cancer cell line MDA-MB-231 via TLR signaling pathway using immunocytochemical technique. MDA-MB-231 breast cancer cells were cultured in RPMI-1640 medium containing 10% FBS, 1% L-glutamine and 1% penicillin/streptomycin. Anti-TLR2, anti-TLR4, anti-MyD88, anti-NFkB, anti-IL-6 and anti-ERK primary antibodies were used for indirect immunohistochemistry after 24 h administrations of Paclitaxel, Metformin and combination of them. The mean values of the staining intensities (mild, moderate, strong and very strong) and percentage of positively stained cells were calculated using H-Score. The results showed that the immunoreactivities of TLR-2, TLR-4, MyD88, NFkB and ERK is increased after the drug treatments while the immunoreactivity of IL-6 has not changed between control and treated groups. To conclude that paclitaxel, metformin and combined therapies on breast cancer cells caused the activation of the TLR-MyD88-ERK signaling pathway which mediates tumor growth and progression, metastasis and drug resistance

The identification of small supernumerary marker chromosomes; the experiences of 15,792 fetal karyotyping from Turkey.

Small supernumerary marker chromosomes (sSMCs) are often associated with developmental abnormalities and malformations are de novo in similar to 60% of the cases. Fluorescence in situ hybridization (FISH) techniques using various probes provided the possibility to analyze and characterize sSMCs. which is highly important for prenatal diagnosis and genetic counseling. We now present the establishment of a specific strategy to identify the origin and structure of the sSMCs using a combination of conventional banding and classical FISH techniques. Based on this strategy, in a series of 15,792 prenatal karyotypes, 20 cases with sSMCs (prevalence 1.26 per 1000) were diagnosed. Eighteen of these cases were completely analyzed by FISH using commercial probes and Chromoprobe Multiprobe (R)-I System. Out of 20 sSMCs 12 were satellited (10 bisatellited and two monosatellited) (60%) and eight were non-satellited (six ring-like and two isochromosomes) (40%). sSMCs were mostly derived from chromosome 15 (10/20) (50%). Euchromatin material was found in 13 cases by various banding and FISH techniques, while in six of 20 sSMCs there was no evidence of euchromatin material. Parental karyotypes could be evaluated in 15 cases and familial inheritance was found in only three of them (20%). We conclude that the proposed strategy for the identification and characterization of sSMCs is accurate and represents a good alternative to novel FISH techniques for modestly equipped cytogenetic laboratories. (c) 2005 Elsevier SAS. All rights reserved

Push and Pull Factors of Why Medical Students Want to Leave Türkiye: A Countrywide Multicenter Study

Phenomenon: Physician immigration from other countries is increasing as developed countries continue to be desirable destinations for physicians; however, the determinants of Turkish physicians’ migration decisions are still unclear. Despite its wide coverage in the media and among physicians in Türkiye, and being the subject of much debate, there is insufficient data to justify this attention. With this study, we aimed to investigate the tendency of senior medical students in Türkiye to pursue their professional careers abroad and its related factors. Approach: This cross-sectional study involved 9881 senior medical students from 39 different medical schools in Türkiye in 2022. Besides participants’ migration decision, we evaluated the push and pull factors related to working, social environment and lifestyle in Türkiye and abroad, medical school education inadequacy, and personal insufficiencies, as well as the socioeconomic variables that may affect the decision to migrate abroad. The analyses were carried out with a participation rate of at least 50%. Findings: Of the medical students, 70.7% had emigration intentions. Approximately 60% of those want to stay abroad permanently, and 61.5% of them took initiatives such as learning a foreign language abroad (54.5%) and taking relevant exams (18.9%). Those who wanted to work in the field of Research & Development were 1.37 (95% CI: 1.22–1.54) times more likely to emigrate. The push factor that was related to emigration intention was the “working conditions in the country” (OR: 1.89, 95% CI: 1.56–2.28) whereas the “social environment/lifestyle abroad” was the mere pull factor for the tendency of emigration (OR: 1.73, 95% CI: 1.45–2.06). In addition, the quality problem in medical schools also had a significant impact on students’ decisions (OR: 2.20, 95% CI: 1.83–2.65). Insights: Although the percentage of those who want to emigrate “definitely” was at the same level as in the other developing countries, the tendency to migrate “permanently” was higher in Türkiye. Improving working conditions in the country and increasing the quality of medical faculties seem vital in preventing the migration of physicians

Treatment efficacy of ribociclib or palbociclib plus letrozole in hormone receptor-positive/HER2-negative metastatic breast cancer.

Background: Ribociclib, palbociclib and abemaciclib are currently approved CDK4/6 inhibitors along with aromatase inhibitors as the first-line standard-of-care for patients with hormone receptor-positive, HER2-negative metastatic breast cancer. Methods: The authors report retrospective real-life data for 600 patients with estrogen receptor- and/or progesterone receptor-positive and HER2-negative metastatic breast cancer who were treated with ribociclib and palbociclib in combination with letrozole. Results & conclusion: The results demonstrated that the combination of palbociclib or ribociclib with letrozole has similar progression-free survival and overall survival benefit in real life for the patient group with similar clinical features. Specifically, endocrine sensitivity may be a factor to be considered in the treatment preference