Introduction: Angiogenesis (neovascularization), which means new vessel construction, is normal and physiologically, wound healing, embryogenesis, a necessary menstrual cycle it's a mechanism. When taken in appropriate amounts together with or separately with nutrients, mucosal and by regulating systemic immunity, ensuring nutritional and microbial balance in the intestines living nonpatogenic microorganisms that positively affect the health of the host it is called "probiotics". Lactic acid bacteria, the most probiotic microorganisms it constitutes its important group. Where probiotics have an effect on angiogenesis, and it is thought to help heal wounds through the road. With this research indicated that roles of Lactobacillus acidophilus and Lactobacillus rhamnosus on angiogenesis if present to demonstrate in vitro methods and the gene expression responsible for the formation of these effects it is intended to reveal. Material and Method: This study is an experimental study conducted in vitro human umbilical cord vein endothelial cell (HUVEC) MTT test in cell culture with (3-[4,5-Dimethylthiazole-2-yl]-2,5-diphenyltetrazolium bromide; Thiazolyl blue) evaluation of viability and proliferation wound healing model, tube formation method and gene expression with real rime-polymer chain reaction (RT-PCR) methods of appointment were used. Results: HUVEC cells L. acidophilus 10⁹ CFU\ml after extract application statistical of mRNA expression of VEGF and FGF genes by control group 24 per hour it was found to increase significantly. L. rhamnosus 10 6 CFU\ml and 109 CFU\ml after application of extracts VEGF gene mRNA by control group 24 per hour its expression was found to be statistically significantly increased. Also L. rhamnosus extracts cell proliferation and migration of in vitro wound model it was found to increase statistically significantly. Conclusion: In this study, in vitro L. acidophilus 10⁹ CFU\ml extract and 10⁶ CFU\ml and 10⁹ CFU\ml extract of L. rhamnosus, VEGF gene mRNA revealed to be effective on angiogenesis in HUVEC cells by increasing expression it is.

The increasing antibiotic resistance in Pseudomonas aeruginosa, responsible for both community-acquired and hospital-acquired infections, is of global significance. The primary mechanisms contributing to resistance development in P.aeruginosa include the increased activity of efflux pumps, decreased permeability of outer membrane porins and the production of carbapenemases. This study aimed to determine the effects of resistance nodulation division (RND) efflux pumps, outer membrane porin D (OprD) outer membrane protein and carbapenemase production on the development of resistance to different antibiotics in P.aeruginosa isolates. Eighty P.aeruginosa isolates obtained from clinical samples in our hospital between 2019 and 2021 were included in the study. Species-level identification of the isolates was performed using MALDI-TOF MS (Bruker Daltonics, Germany). Antibiotic susceptibilities were determined using the VITEK (R) 2 (bioM & eacute;rieux, France) system according to the criteria of the European Committee on Antimicrobial Susceptibility Testing (EUCAST). The expression levels of the outer membrane porin protein OprD and the regulatory genes of efflux pumps (mexB, mex C, mex E, and mex X ) were investigated using real-time quantitative reverse transcriptase polymerase chain reaction (Rt-qPCR). The rpsL gene was used as the reference gene and P.aeruginosa PAO1 strain was used as the control strain in Rt-qPCR. Comparative expression analysis was calculated using the delta-delta cycle threshold (Delta Delta Ct) method. The presence of bla(OXA-48), bla(NDM), bla(VIM), bla(IMP), bla(KPC) and bla(OXA-10) was investigated by PCR. Clonal relationships among the isolates were determined by AP-PCR using the M13 primer. Band profiles were analyzed using GelCompar II (Applied Maths) software. Decreased OprD expression was detected in 63.7% of the isolates and 74% of carbapenem-resistant isolates. The decrease in OprD expression was found to be significant only between the carbapenem-susceptible (n= 30) and carbapenem-resistant (n= 50) groups (p= 0.014). The overexpression rate of mexB was 82% in carbapenem-resistant isolates and 33% in carbapenem-susceptible isolates; 76.6% in multidrug-resistant (MDR) isolates and 45.5% in non-MDR isolates (p= 0.001 and p= 0.004, respectively). The overexpression rate of mexX was 68% and 40% in the amikacin-resistant and susceptible groups, respectively; and 64.3% and 40.4% in the gentamicin-resistant and susceptible groups, respectively (p= 0.02 and p= 0.041, respectively). Over- expression of mexX was found in 61.7% of MDR isolates and 30.3% of non-MDR isolates (p= 0.006). No significant difference was found in the expression of mexC and mexE between antibiotic-susceptible and resistant groups. While bla OXA-10 was detected in 32% (16/50) of carbapenem-resistant isolates, bla OXA-48 , bla NDM , bla VIM , bla IMP , bla KPC were not detected in any of the isolates. AP-PCR analysis identified 64 different genotypes and no dominant genotype was observed. The isolates were grouped into 14 different clusters with a clustering rate of 36%. RND efflux systems play a crucial role in the development of antibiotic resistance in P.aeruginosa. This study showed that overexpression of mexB and decreased expression of OprD contributed to the development of carbapenem resistance, while overexpression of mexX contributed to the development of amikacin and gentamicin resistance. Overexpression of mexB and mexX has significant correlation with MDR isolates. The detection of bla OXA-10 in 16 isolates suggests that the presence of this resistance gene may contribute to the development of carbapenem resistance.

Investigation of the Effects of Polyunsaturated Fatty Acid Ratios in Human SH-SY5Y Cells by in vitro Methods - Turkish Journal of Pharmaceutical Sciences

Objective: The mitochondria isolation method is used to investigate mitochondrial processes. RT-qPCR measurement of gene transcription levels is frequently used in these studies. In this method, it is important that the housekeeping genes used in the normalisation of the results are stable. The study aimed to determine which of two housekeeping genes is more stable in isolated mitochondria. Material and Method: Mitochondria were isolated from FHC cells. Membrane integrity and functionality were measured by mitotracker and JC-1 stainings and ROS ratio by flow cytometry. Housekeeping genes GAPDH and β-actin were used in RT-qPCR, and gene stability was calculated using ΔCq method and percentage coefficient of variance. For gene validation, CAT and SOD1 transcriptions were calculated by the 2-ΔΔCq method. These genes were selected because oxidative stress status has been detected. Result and Discussion: Isolated mitochondria membrane integrity was preserved and the membrane potential ratio was found 91%. ROS ratio was 1.6% and there was no oxidative stress that would affect the antioxidant enzymes levels. ΔCq values were 4.54±0.06 for β-actin and 0.69±0.16 for GAPDH, with 1.33% and 23.95% percentage coefficient of variance respectively. CAT transcription levels were 25.7±2.6 and 1.8±0.33, while SOD1 levels were 70.1±12.7 and 4.8±0.6 correspondingly. It was concluded that β-actin may be more stable than GAPDH. The potential impact of housekeeping gene selection on outcomes should be considered.

Objectives: The Pasteur effect in cellular energy metabolism is defined as the switch to anaerobic glycolysis due to alterations in ambient oxygen concentration or inhibition of the electron transport chain (ETC) and oxidative phosphorylation. This study aimed to determine the effect of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) infection on the energy metabolism of leukocytes among Coronavirus Disease 2019 (COVID-19) patients at various stages. Methods: A total of 90 cases were analyzed in asymptomatic, mild, moderate, and severe COVID-19 patients. Levels of aerobic glycolysis intermediates and products of anaerobic glycolysis were measured in leukocytes with liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. Results: A decrease in the rate of aerobic glycolysis was found as the disease progressed (p-values<0.001), while the anaerobic glycolysis rate was unchanged in all study groups because lactate levels had no difference between asymptomatic, mild, moderate, and severe groups (p=0.754). Moreover, the ratios of lactate to Tricarboxylic Acid (TCA) cycle parameters demonstrated a significant reduction in the rate of aerobic glycolysis of COVID-19 patients at different stages (p-values<0.001). Furthermore, Receiver Operating Characteristic (ROC) curve analysis for citrate, alpha-ketoglutarate, fumarate, lactate, lactate/citrate, lactate/alpha-ketoglutarate, and lactate/fumarate had significant cut-off values in Moderate and Severe patients (p<0.001). Conclusions: Our results suggest that the Pasteur effect from aerobic to anaerobic glycolysis was shown to be induced as the severity of COVID-19 progresses.