The results of a close follow-up of trainees to gain a good blood collection practice

BackgroundPhlebotomy is one of the most important steps in the preanalytical phase of a clinical laboratory process. In order to decrease phlebotomy errors, this specific procedure should be taught in detail by laboratory organizations. Our study aims to practice the training program on venous blood sampling and observe the close follow-up results.MethodsIn this observational study, 127 students who started their summer internship in Antalya Education and Research Hospital were given a one-day theoretical phlebotomy training in accordance with the Venous Blood Sampling Guidelines. After the theoretical training, phlebotomy applications of 10 students who were working in the field of out-patient blood sampling were observed both with and without their knowledge. A comprehensive checklist related to phlebotomy was created by the trainers in Antalya Education and Research Hospital and the observers answered each question as yes or no. For the statistical analysis, IBM SPSS Statistics 21.0 was used.ResultsAfter the theoretical education, the trainees were observed but no significant difference was found between the first and the second informed observations (p = 0.125). The students were observed three times more in the following week without their knowledge. There was a statistically significant difference between the first and the third unannounced observations (p=0.001).ConclusionsIn order to perform phlebotomy correctly, apart from theoretical education, a close follow-up is necessary too.     

Chitosan and silver nanoparticles are attractive auxin carriers: A comparative study on the adventitious rooting of microcuttings in apple rootstocks

Nanocarriers for encapsulation and sustained release of agrochemicals such as auxins have emerged as an attractive strategy to provide enhanced bioavailability and efficacy for improved crop yields and nutrition quality. Here, a comparative study was conducted on the effectiveness of chitosan-as a biopolymeric nanocarrier- and silver-as a metallic nanocarrier- on in vitro adventitious rooting potential of microcuttings in apple rootstocks, for the first time. Auxins indole-3-acetic acid (IAA) and indole-3-butyric acid (IBA) loaded silver (nAg) or chitosan nanoparticles (nChi) were synthesized. Scanning electron microscopy and transmission electron microscopy studies showed the spherical shape of the nanoparticles. The average particle size of IAA-nChi was 167.5 ± 0.1 nm while that of IBA-nChi was 123.2 ± 2.6 nm. The hydrodynamic diameter of the nAg-IAA and nAg-IBA particles were measured as 93.66 ± 5 nm and 71.41 ± 3 nm, respectively. Fourier transform infrared spectroscopy analyses confirmed the encapsulation of IAA or IBA in the chitosan nanoparticles. Meanwhile, the characteristic peaks of IAA or IBA were detected on silver nanoparticles. In-vitro adventitious rooting of microcuttings of Malling Merton 106 (MM 106) was significantly higher both in chitosan and silver nanoparticles loaded with IAA or IBA (91.7%–62.5%) compared to free IAA or IBA applications (50.0%–33.3%), except for 2.0 mg L^–1 IBA (66.7%). However, the application of 2 mg L^–1 IBA and IBA-nChi at all concentrations caused an undesirable large callus development.     

Direct and Indirect Control of the Initiation of Meiotic Recombination by DNA Damage Checkpoint Mechanisms in Budding Yeast

Meiotic recombination plays an essential role in the proper segregation of chromosomes at meiosis I in many sexually reproducing organisms. Meiotic recombination is initiated by the scheduled formation of genome-wide DNA double-strand breaks (DSBs). The timing of DSB formation is strictly controlled because unscheduled DSB formation is detrimental to genome integrity. Here, we investigated the role of DNA damage checkpoint mechanisms in the control of meiotic DSB formation using budding yeast. By using recombination defective mutants in which meiotic DSBs are not repaired, the effect of DNA damage checkpoint mutations on DSB formation was evaluated. The Tel1 (ATM) pathway mainly responds to unresected DSB ends, thus the sae2 mutant background in which DSB ends remain intact was employed. On the other hand, the Mec1 (ATR) pathway is primarily used when DSB ends are resected, thus the rad51 dmc1 double mutant background was employed in which highly resected DSBs accumulate. In order to separate the effect caused by unscheduled cell cycle progression, which is often associated with DNA damage checkpoint defects, we also employed the ndt80 mutation which permanently arrests the meiotic cell cycle at prophase I. In the absence of Tel1, DSB formation was reduced in larger chromosomes (IV, VII, II and XI) whereas no significant reduction was found in smaller chromosomes (III and VI). On the other hand, the absence of Rad17 (a critical component of the ATR pathway) lead to an increase in DSB formation (chromosomes VII and II were tested). We propose that, within prophase I, the Tel1 pathway facilitates DSB formation, especially in bigger chromosomes, while the Mec1 pathway negatively regulates DSB formation. We also identified prophase I exit, which is under the control of the DNA damage checkpoint machinery, to be a critical event associated with down-regulating meiotic DSB formation.     

Determination of plasma microRNA for early detection of gastric cancer

Gastric cancer is the fourth most prevalent malignancy worldwide and remains the second most common cause of cancer-related death globally. Understanding the molecular structure of gastric carcinogenesis might identify new diagnostic and therapeutic strategies for this disease. Thus, early detection of gastric cancer is a key measure to reduce the mortality and improve the prognosis of gastric cancer. There have recently been several reports that microRNAs (miRNAs) circulate in highly stable, cell-free forms in blood. Because serum and plasma miRNAs are relatively easy to access, circulating miRNAs also have great potential to serve as non-invasive biomarkers. Although a number of miRNAs associated with gastric cancer have been identified, the underlying mechanism of these miRNAs in tumorigenesis and tumor progression remains to be investigated. The purpose of this study is to identify the potential of serum miRNAs as biomarkers for early detection of gastric cancer patients. RNA was isolated using the High Pure miRNA Isolation Kit (Roche) following the manufacturer’s protocol. cDNA and preamplification protocols were obtained from the isolated plasma miRNAs. The BioMark? 96.96 Dynamic Array (Fluidigm Corporation) for real-time qPCR was used to simultaneously quantite the expression of 740 miRNAs. All statistical analyses were performed using the Biogazelle’s qbase PLUS 2.0 software. In this study, among 740 miRNAs that we analyzed only miR-195-5p was significantly (p < 0.05, fold changes = 13, 3) down-regulated in gastric cancer patients compared with control. We demonstrated that miR-195-5p is a novel tumor suppressor miRNA and may contribute to gastric carcinogenesis. The miRNA expression profile described in this study should contribute to future studies on the role of miRNAs in gastric cancer.     

Development of Vaccine Prototype Against Zika Virus Disease of Peptide-Loaded PLGA Nanoparticles and Evaluation of Cytotoxicity

Zika virus has recently evolved from an obscure mosquito-borne pathogen to an international public health concern. People with Zika virus disease can have indications including mild fever, skin rash, conjunctivitis, muscle pain, malaise or headache. Effective vaccines are needed for controlling and preventing the disease. In the current study, we aim to design the substructure for vaccine against Zika virus by forming antigenic peptide epitope of the disease. Zika peptide loaded poly (lactic-co-glycolic acid) (PLGA) nanoparticles have been fabricated in the present work as a potential artificial vaccine. UV and FT-IR Spectrophotometers and ZetaSizer were used for studying the nanoparticles, and Scanning Electron Microscope was used for morphological examination. The nanoparticles (NPs) yield, encapsulation efficiency, the peptide loading capacity were determined and in vitro release of the peptide was studied. Cytotoxic effects of the various concentrations of Zika peptide, blank PLGA nanoparticles and Zika peptide loaded PLGA nanoparticles on ECV304 human epithelial cells were determined via MTT assay. The present paper could be considered as one of the important steps in the use of nanoparticles for constructing the new generation of vaccination systems.

     

Basal metabolic rate and autonomic nervous system dysfunction in men with spinal cord injury

Objectives: The purpose of this study was to determine the relationship between autonomic nervous system dysfunction and basal metabolic rate (BMR), and the effect of spasticity on basal metabolic rate. Research Method and Procedures: Twenty men (11 paraplegic and 9 tetraplegic) with American Spinal Injury Association (ASIA)‐A and ‐B grade chronic spinal cord injury (SCI) participated in this study. Total body fat mass and lean tissue mass were measured in all participants using DXA by standard methods. Patients were allocated into 2 groups to determine the effect of autonomic nervous system dysfunction on BMR: Group I (T6 and upper‐level injuries with history of autonomic dysreflexia) and Group II (T7 and lower‐level injuries without history of autonomic dysreflexia). Measurements of BMR were determined by indirect calorimetry under standardized conditions. Results: There were 13 patients in Group I and 7 patients in Group II and the difference between these two in terms of time since injury, BMI, age, weight, lean tissue mass, BMR, and BMR/kg were not significant. Conclusion: We concluded that autonomic nervous system dysfunction does not affect BMR, and it might be ignored in considering energy needs in spinal cord injury.     

Antioxidant and antimicrobial actions of the clubmoss <Emphasis Type="Italic">Lycopodium clavatum</Emphasis> L.

Purpose of the present study was to evaluate antioxidant, antibacterial, antifungal, and antiviral activities of the petroleum ether, chloroform, ethyl acetate and methanol extracts as well as the alkaloid fraction of Lycopodium clavatum L. (LC) from Lycopodiaceae growing in Turkey. Antioxidant activity of the LC extracts was evaluated by 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical-scavenging method at 0.2 mg/ml using microplate-reader assay. Antiviral assessment of LC extracts was evaluated towards the DNA virus Herpes simplex (HSV) and the RNA virus Parainfluenza (PI-3) using Madin-Darby Bovine Kidney (MDBK) and Vero cell lines. Antibacterial and antifungal activities of the extracts were tested against standard and isolated strains of the following bacteria; Escherichia coli, Pseudomonas aeruginosa, Proteus mirabilis, Acinobacter baumannii, Klebsiella pneumoniae, Staphylococcus aureus, Bacillus subtilis as well as the fungi; Candida albicans and C. parapsilosis. All of the extracts possessed noteworthy activity against ATCC strain of S. aureus (4 μg/ml), while the LC extracts showed reasonable antifungal effect. On the other hand, we found that only the chloroform extract was active against HSV (16–8 μg/ml), while petroleum ether and alkaloid extracts inhibited potently PI-3 (16–4 μg/ml and 32–4 μg/ml, respectively). However, all of the extracts had insignificant antiradical effect on DPPH. In addition, we also analyzed the content of the alkaloid fraction of the plant by capillary gas chromatography-mass spectrometry (GC-MS) and identified lycopodine as the major alkaloid.     

Alterations of oxidative-antioxidative status in human cutaneous leishmaniasis

Enhanced lipid peroxidation and decreased antioxidant defences have been defined in several diseases. In the present study, we aimed to evaluate the oxidative-antioxidative status of patients with cutaneous leishmaniasis (CL). Concentrations of erythrocyte lipid peroxidation (LPO), as an indicator for the oxidative status, reduced glutathione (GSH), glutathione peroxidase (GSH-Px) and serum vitamin C levels, as indicators for the antioxidative status, were measured. Seventy patients aged between 15 and 50 years (38 patients had active CL and 32 patients had healed CL) and 40 healthy controls aged between 19 and 50 years were included in the study. LPO and GSH of the patients with active CL were significantly higher (p < 0.001), whereas erythrocyte GSH-Px and serum vitamin C levels were lower (p < 0.001, p < 0.01 respectively) than those of healthy controls. There was a significant inverse correlation between LPO and serum vitamin C level (r=-0.32, p < 0.05) in active CL. No significant correlation of LPO, GSH, GSH-Px and serum vitamin C levels in control groups or in the group with healed CL was detected. In the light of our findings it is possible to conclude that patients having CL are affected by oxidative stress, which most likely induces the endogenous antioxidant system. An imbalance between the oxidant and antioxidant systems occurs and the suppressed antioxidants and increased lipid peroxidation may contribute to the progression of the disease.