Volume 47, Issue 4

The increasing availability and use of POCT are influenced by many factors, including the industry trend toward patient-centered care and decentralization of health care; the increasing prevalence of infectious diseases, which includes the current use of rapid SARS-CoV-2 testing; the increasing incidence of lifestyle diseases such as diabetes, heart disease, and hypertension; and advances in in-vitro medical diagnostics. The use of POCT can increase the efficiency of services and improve patient outcomes. However, the variability of the testing environment and conditions, as well as the competency of the staff performing the tests, can have a significant impact on the quality and accuracy of POCT results. Most personnel performing POCT are not trained laboratory staff and may not be as familiar with the processes associated with testing, such as patient preparation, specimen collection, management of equipment and supplies, calibration and maintenance of equipment, the performance of the test, quality control, interpretation of results, and reporting/documentation of results related to the particular patient. Therefore, personnel performing POCT must have the appropriate training and experience to ensure that test results are accurate and reliable. This review outlines the specific personnel training requirements based on international standards that must be considered to ensure the quality of test results and describe the competency criteria required for POCT compliance.

Objectives Microorganisms produce optically single bioactive natural products; the process is cheap as compared to chemical synthesis and environmentally friendly. Geodin 1 was isolated from the soft coral-derived fungus Aspergillus sp. It has a broad range of biological activities such as antiviral, antimicrobial, glucose stimulator for rat adipocytes, enhancement of fibrinolytic and cytotoxic activities, and is a subunit of the first nonpeptide and inhibitory active galanin receptor. In this work, we studied the effects of sodium chloride (NaCl) salt concentration on the production of geodin 1 and improved its yield to a multi-gram quantity through media optimization from a marine-derived fungus Aspergillus sp. Methods The fungal strain was cultivated at various concentrations of NaCl salt in rice medium and extracted after different intervals of time. Results The yield of geodin 1 was 137.2 mg/L at optimal conditions. The optimal conditions for the high yield of geodin 1 were found as rice medium with 2.0% NaCl salt and 3 weeks incubation at room temperature. Conclusions The concentration of NaCl salt greatly affects the yield of geodin and hence its biosynthetic pathway.

Objectives As one of the most widely used tests, the erythrocyte sedimentation rate (ESR) is a measurement of sedimentation reaction in blood samples. Although the recommended method for ESR measurement is the Westergren method, this method has some disadvantages in comparison with automated ESR systems. In this cross-sectional study, we aimed to compare Vision C with the Westergren method. Methods The Vision C automated ESR system and the modified Westergren method were compared using K 3 EDTA-containing tubes and citrated blood tubes from randomly selected 100 patients. Precision, bias, and interference calculations were performed. Results The mean difference between the Vision C (room temperature) and the modified Westergren method was 22.8 ± 26.6 mm/h (95% CI for the mean was 17.50–28.08 mm/h). The mean difference between the Vision C (18 °C) and the modified Westergren method was 27.4 ± 30.6 mm/h (95% CI for the mean was 21.3–33.5 mm/h). The calculated regression analysis equation was “ y =  0.263  +  1.053x ” and “ y = − 0.530  +  0.851x ” for the Vision C (room temperature) and the Vision C (18 °C), respectively. The imprecision values ranged at 7.55–17.09%. According to our external quality control results, bias was 11.11 and 9.66 for the low- and high-level samples, respectively. Conclusions The Vision C automated ESR system has a comparable analytical performance with the modified Westergren method. The Vision C automated system may be utilized in ESR measurements with quality control evaluations. Nevertheless, temperature correction using Manley’s monogram causes an important negative bias and should be taken into consideration during the evaluation of the Vision C results.

Objectives Acute kidney injury (AKI) is a critical clinical event characterized by a reduction in the excretory function of the kidneys. N-acetylcysteine (NAC), N-acetylmethionine (NAM) and N-acetylglucosamine (NAG) are antioxidants with scanty known genetic mechanisms. We aimed to assess both kidney injury molecule-1 (KIM-1) and growth-arrested DNA damage-inducible gene-153 (GADD-153) genes expression in paracetamol (PA) induced AKI. Also, to recognize whether NAC, NAM and/or NAG have roles in altering the expression of these genes for ameliorating the AKI induced by PA. Methods The present preliminary study achieved the AKI model by oral administration of PA therapeutic dose for 15 days in experimental male rats. Serum urea, creatinine, and renal oxidative stress parameters were analyzed. Genetic expression of KIM-1 and GADD-153 were determined using real time-PCR. Results Significant elevations of serum urea, creatinine and nitric oxide in renal tissue after PA administration; however, total thiol content was reduced. In addition, both KIM-1 and GADD-153 were upregulated. These biochemical alterations were improved after using NAC and partially after NAM; however, NAG had little effect. Conclusions Up-regulation of both KIM-1 and GADD-153 occur in AKI induced by PA, which was significantly reversed by NAC.

Objectives Breast cancer is the leading cause of mortality in today’s world. An alarming rise in cancer incidence has been observed in the South Asian region. The aberrant molecular mechanisms regulating cell proliferation and development contribute to cancer development. A better understanding of the detailed molecular mechanisms at genetic and epigenetic levels can help to treat breast cancer more efficiently. The present study is aimed to identify the possible association of MAP3K1 SNP rs889312 and MAP3K9 rs11628333 in breast cancer in the South Asian region. Methods Female breast cancer patients were recruited in the study. DNA was isolated from the blood samples collected from the patients. PCR-RFLP was used for genotyping, and data analysis was done by SPSS software. Results Genotyping data for MAP3K1 SNPrs889312 showed statistically significant association with breast cancer, while MAP3K9 SNPrs11628333 showed characteristic association of rare allele heterozygote’s and homozygotes in pre and post-menopausal patients, respectively. Conclusions The study concludes a strong association of the rs889312 with breast cancer in the Pakistani population and a characteristic association of unique genotypes TC and CC in pre- and post-menopausal breast cancer patients. These findings can provide a ready tool as a breast cancer marker in south Asian populations.

Objectives Adipose tissue mediates various bioactive molecules and cytokine discharge. The anti-inflammatory cytokine, interleukin-10 (IL-10), has roles in systemic inflammation. Matrix metalloproteinases (MMPs) are endopeptidases implicating in tissue remodeling, and extracellular matrix degradation. Interleukins and MMPs may have specific roles in obesity development. In this investigation, we marked the roles of IL-10, MMP-2, and MMP-9 in obesity and its related clinical phenotypes. Methods Using real-time quantitative polymerase chain reaction (RT-qPCR), also ELISA, IL-10, MMP-2, and MMP-9 mRNA and protein levels were detected respectively in the subcutaneous adipose tissues of 34 patients with obesity and 36 healthy individuals. Results MMP-2 and MMP-9 gene expression were significantly downregulated in obesity patients compared to controls (p=0.004, p=0.045). Nevertheless, IL-10 was elevated in the obesity group as to controls (p=0.010). MMP-2 mRNA expression was correlated with fasting blood glucose levels (r=0.426, p=0.013) in the patient group. As for protein levels, MMP-2 concentration decreased in patients compared to controls (p=0.001). Moreover, MMP-2 was correlated with BMI (r=−0.411; p=0.022) and weight (r=−0.381; p=0.034) in obesity group. Conclusions MMP-2 , MMP-9, and IL-10 may be related to increased susceptibility to obesity development and its related phenotypes in a sample of Turkish patients with obesity.

Objectives This study aimed to examine the role of miR-221, miR-650, and miR-4534 expressions in the development, pathogenesis, and early diagnosis of prostate cancer. Methods The study included 83 participants: 37 patients with PCa, 31 patients with BPH, and 15 healthy subjects. MiRNA expressions in plasma samples was evaluated by quantitative RT-PCR. Results Plasma miR-221 and miR-4534 levels were significantly upregulated in the PCa and BPH groups compared to the control group. A significant difference was determined between the presence of lymph node metastasis and the expressions of miRNAs. In the ROC analysis of the miRNAs, it was determined that the AUC for miR-221 was 0.737 with a sensitivity of 57% and specificity of 100%, AUC for miR-650 was 0.706 with a sensitivity of 62% and specificity of 93% and AUC for miR-4534 was 0.800 with a sensitivity of 73% and specificity of 93%. Conclusions Overexpression of miR-221, miR-650, and miR-4534 may distinguish PCa and BPH from healthy controls, but seems to be insufficient in differentiating PCa from BPH when used alone or in combination. However, these oncogenic miRNAs may have a role in determining the development and progression of the disease by suppressing the tumor suppressor genes they target.

Objectives Aim of the study was to reveal new biomarker genes to target breast cancer stem-like cells (BCSC-like) and then sensitize BCSC-like cells to chemotherapy by silencing MDR1 gene found to be the most suitable target. Methods Drug resistance associated genes were screened by cDNA microarray to unveil biomarker genes in drug resistance breast cancer model cells. Drug resistance was then reversed by silencing MDR1 gene in BCSC-like cells. The effect of silencing was monitored by real-time cell proliferation analysis. Differential expressions of MDR1 , ALDH1A3 , EGFR and BAG4 genes were identified by real-time PCR. P-glycoprotein (P-gp) expression level and its activity were investigated by Western blot and flow cytometry measurements, respectively. Results 16 new biomarker genes were identified upon gene expression analysis by cDNA microarray. MDR1 gene was selected as the most potent target gene and silencing of it caused down-regulation of MDR1 , ALDH1A3 , EGFR , BAG4 expression and P-glycoprotein activity and expression in BCSC-like cells. At the end, silenced BCSC-like cells were found to be more responsive to paclitaxel therapy. Conclusions In conclusion, si MDR1 silencing is an effective way to reverse multidrug resistance and malignancy. New biomarker genes revealed in this study require to be investigated to target stemness of BC.

Objectives We aimed to determine the relationship of Lyso-Gb1 levels, bone biomarkers, and mutation findings with bone marrow burden (BMB) scores. Methods Lyso-Gb1 and bone biomarkers, and BMB scores of 10 Gaucher type 1 (GD1) patients under enzyme therapy were prospectively evaluated. Results Ten GD1 patients, aged between 4.5 and 40 (mean 23 ± 11 years), were included in the study. Four patients were homozygous for L444P/L444P, and six patients were compound heterozygous for N370S/R415H. We found positive correlations between pain and BMB scores with Lyso-Gb1 levels (r=0.889, p=0.001 and r=0.701, p=0.035, respectively). There were negative correlations between bone mineral density (BMD) of both the lumbar spine and femoral neck between Lyso-Gb1 levels (r=−0.929, p=0.001 and r=−0.893, p=0.007, respectively). Patients with L444P/L444P mutation had higher Lyso-Gb1 levels and BMB, pain scores and lower BMD measurements than patients with N370S/R415H (p=0.01, p=0.02, p=0.03, p=0.04, p=0.04, respectively). Conclusions There was an apparent correlation between, Lyso-Gb1 levels, BMB scores and genotype in evaluating bone involvement in Gaucher patients.

Objectives Endoplasmic reticulum aminopeptidase-1 ( ERAP1 ) polymorphic changes cause autoimmunity. To understand the contribution of ERAP1 to the occurrence of rheumatoid arthritis (RA) disease, we investigated the relationship between ERAP1 and RA. Methods This study was conducted with 201 patients and 171 healthy controls. The rs26653, rs27044, rs27582, rs28096, and rs30187 polymorphic regions of ERAP1 were investigated. The comparison was done with Arlequin software and logistic regression. Haplotypes were analyzed with Phylogenetic Network software. ERAP1 was modeled using Promod3. Topological changes in ERAP1 were analyzed with TM-Score. Results The results showed that rs26653G>C (p=0.002, OR=2.001, 95%CI=1.276–3.137), rs27044C>G (p=0.037, OR=1.583, 95%CI=1.028–2.440), rs27582G>A (p<0.05, OR=0.348, 95%CI=0.194–0.622) and rs30187C>T (p=0.006, OR=1.849, 95%CI=1.191–2.870) polymorphisms are associated with RA disease risk. The relationship between rs28096 polymorphism and RA disease risk could not be determined (p=0.509). The risk haplotype for rheumatoid arthritis was determined as [CGAAT]. It was determined that polymorphisms of ERAP1 cause changes in the entry pocket of substrate and ligand. Conclusions We report a haplotype [CGAAT] that is associated with RA risk from Turkey that has not been described before. These data will make important contributions to elucidating the molecular mechanism of RA.

Objectives In this study, we aim at deciphering the phenolic content of Vitex agnus-castus L. leaf and fruit extracts prepared with different methods and relate it to their antioxidant activity. Methods In this study, phenolic compounds and the antioxidant potential of the ethanol fruit and leaf extracts of V. agnus-castus L. (Chaste tree) were evaluated spectrometrically. Furthermore, selected polyphenols, i.e., chlorogenic acid and rutin, were determined by the HPLC-DAD method qualitatively and quantitatively. Results The results obtained from leaf and fruit extracts were compared with a commercial product (CP) containing the fruit extract of V. agnus-castus . Leaf extract was found to be richer in flavonoids when compared to the fruit counterparts. Accordingly, they also showed higher antioxidant activity. Conclusions Extracts prepared here can be considered as promising antioxidant agents for future therapeutic formulations.

Objectives The majority of research programs had been focused on extract from the fruiting body. However, fungal mycelium obtained by submerged cultivation also has higher pharmacological potential. The aim of this study was investigation of the antioxidant and antimicrobial potential of Ganoderma lucidum and Trametes versicolor fungal mycelium biomass obtained by submerged cultivation and liquid fluid. Methods The antioxidant activity was evaluated by comparing the oxidation kinetics of the reduced form of 2,6-dichlorophenolindophenol with atmospheric oxygen in the presence/absence of biological samples. A criterion for evaluating for assessing the antioxidant activity was the values of the inhibition constant of the oxidation reaction. Antimicrobial activity of the fungal extracts was performed by agar disc diffusion method. Results A comparative analysis of antioxidant potential of G. lucidum and T. versicolor mycelium biomass and liquid fluid was conducted for the first time. It was found that antioxidant activity of fungal biomass depends on the cultivation time. The most active were the fractions obtained on the 21st day of fungal cultivation. Fungal extracts were more effective against Gram-positive bacteria compared to Gram-negative bacteria, micromycete and yeasts. Conclusions The results showed that submerged cultivation of mushrooms has significant industrial potential.

Objectives Patients suffering from myocardial ischemia-reperfusion (IR) injuries usually have varying degrees of negatively-affecting comorbidities like hyperlipidemia. We evaluated the preconditioning effect of ginsenoside-Mc1 on reperfusion injury-induced myocardial arrhythmias, along with cardiac oxidative stress, endoplasmic reticulum stress protein expression, and histological damage in hyperlipidemic rats, and further, explore the role of JAK2/STAT3 activity. Methods Thirty-five Sprague–Dawley rats fed a high-fat diet for eight weeks. Ginsenoside-Mc1 (10 mg/dL, IP) was administered to hyperlipidemic rats daily for one month before IR injury. IR injury was induced by 35 min LAD coronary artery ligation and subsequent 60-min reperfusion. A selective JAK2 inhibitor (AG490) was injected before IR injury. Electrocardiography was recorded and myocardial arrhythmias (ventricular premature complexes, tachycardia, and fibrillation) were evaluated in the reperfusion phase according to Lambeth convention. Hematoxylin-Eosin staining, spectrophotometry, and Western blotting techniques were employed to measure the endpoints. Results IR injury in hyperlipidemic rats significantly increased the reperfusion-induced myocardial arrhythmias counts, timing, incidence, and severity. The expression of proteins of endoplasmic reticulum stress (p-PERK, p-eIF2α, CHOP), and oxidative stress marker malondialdehyde were significantly upregulated following IR induction, whereas antioxidant superoxide-dismutase and JAK2/STAT3 proteins expression significantly reduced, as compared with untreated-hyperlipidemic rats. Administration of ginsenoside-Mc1 to hyperlipidemic rats significantly corrected the arrhythmogenic feature of IR injury, reduced phosphorylation and expression of PERK, eIF2α, CHOP, and improved oxidative stress and histological changes. Interestingly, inhibition of the JAK2/STAT3 pathway via AG490 significantly abolished ginsenoside-Mc1-induced cardioprotection. Conclusions Taken together, ginsenoside-Mc1 exerts substantial anti-arrhythmogenic influences against myocardial IR injury in hyperlipidemic rats through activation of JAK2/STAT3 pathway and subsequent reduction of oxidative/endoplasmic reticulum stress.

Objectives This study aims the covalent immobilization of Jack bean urease on Fe 3 O 4 magnetic nanoparticles via glutaraldehyde (urease@MNPs-Si-Glu) and epichlorohydrin (urease@MNPs-ECH) spacer arms. Methods The optimum pH and temperature, thermal, storage and reuse stability of free and immobilized urease preparations were investigated. Thermodynamics characterizations of free and immobilized urease preparations were also studied. Results The free urease and both immobilized urease preparations showed maximal catalytic activity at pH 7.5. The free urease had a maximal catalytic activity at 50 °C, while the both immobilized urease preparations exhibited their maximal catalytic activities at 70 °C. The urease@MNPs-Si-Glu and urease@MNP-ECH showed 2.7- and 1.9-fold higher thermal stability than the free urease at 60 °C, respectively. The free urease remained 30% of their initial activity at 4 °C, while urease@MNPs-Si-Glu and urease@MNPs-ECH retained 72 and 60% of their initial activities at the same conditions. The urease@MNPs-Si-Glu and urease@MNPs-ECH preserved 53 and 52% of their initial activities, respectively for urea hydrolysis after 20 reuses. Conclusions The urease@MNPs-Si-Glu and urease@MNPs-ECH may find a potential application area in urea hydrolysis.

Objectives The study was aimed to evaluate the effect of essential oils against fungal pathogens including the chemical analysis (GC-MS) of the most effective essential oil. Methods The antifungal effect of essential oils was assessed after morpho-molecular identification of Alternaria alternata . Mycelial growth inhibition (%) in in vitro treatment of eight essential oils at different concentrations was observed against A. alternata . Fungitoxicity assay was done following chemical composition analysis by Gas chromatography-mass spectrometry. Direct application of thyme oil was performed on healthy fruits against A. alternata . Results All eight essential oils showed mycelial growth inhibition at different levels, whereas thyme oil was found more efficacious against A. alternata . Chemical composition analysis detected 32 components and thymol was found in a higher percentage. Direct application of the most significant concentration of thyme oil was found effective against A. alternata with a varying decrease in decaying percentage. Conclusions Investigation of this study proved thyme oil as a potential and eco-friendly botanical fungicide effectively used against A. alternata on loquat fruit. The current study explored that thyme oil could be potentially used against A. alternata and its compounds could be further investigated for the development of eco-friendly approaches for the control of postharvest fruit rots.

Objectives This study investigated the association between the pre-miRNA146a C > G rs2910164 polymorphism and serum TNF-α in Egyptian patients with IS. Methods A case-control study was conducted on 75 Egyptian cases with IS and 75 sex-matched control subjects aged 57–65 years. Genomic DNA analysis of pre-miRNA146a and TNF-α measurement was performed with real-time PCR and ELISA, respectively. Results There was a statistically significant difference between cases of ischemic stroke (IS) and control subjects in pre-miRNA146a rs2910164 GG genotype (p=0.017) and G allele (p=0.005). The pre-miRNA146a rs2910164 is significantly associated with large artery atherosclerosis [LAA] in GG genotypes (p=0.019) and G alleles (p=0.004) compared to control subjects. There was a highly statistically significant increase in TNF-α levels (p<0.001) in IS compared to the control group. There was also a statistically significant increase in TNF-α levels (p=0.001) in GG genotypes in IS. Conclusions Our results showed that there was a statistically significant association between pre-miRNA146a rs2910164 GG genotype and susceptibility to IS and LAA. In addition, there was a statistically significant association between pre-miRNA146a rs2910164 GG genotype and TNF-α in IS subjects.

Objectives Virtual laboratory simulations (VLSs) are computer-based tools that offer unlimited application options in scientific, medical, and engineering fields. The aim of this study was to evaluate whether VLSs are efficient learning tools and how these simulations can be integrated into laboratory practice in medical laboratory education. Methods In this pre-test/post-test control group study, 32 volunteers were randomly assigned to either experimental or control groups. The experimental group performed laboratory simulations based on biochemistry and microbiology and then completed a self-report survey to evaluate their satisfaction and beliefs about simulations. Results In the experimental group, post-test scores of each simulation were significantly elevated compared to pre-test scores; however, pre- and post-test scores of control group were statistically the same. The experimental group agreed that these simulations should be applied before theoretical lectures and laboratory practices. They also highlighted that translating from English to their native language creates difficulties in applying and understanding the simulation. Conclusions We emphasized that VLSs are excellent learning tools that increase not only the knowledge but also the self-motivation and focus of the students. Based on feedbacks, native language options are necessary to enable the students to achieve equality of opportunity in education.

Objectives The Presidency of the Republic of Turkey Human Resources Office has stated that a Career Planning Course (CPC) should be implemented in higher education curricula by 2020. An institutional CPC consisting of 10-online sessions was designed and implemented in at Ege University Faculty of Medicine (EUFM) curriculum. This study reports the design, implementation, and evaluation of this new CPC at EUFM. Methods A descriptive, cross-sectional research design was used in this study. An online questionnaire (n=253) focusing mainly on Kirkpatrick-model Level 1 was administered to gather students’ feedback on CPC as well as perceptions regarding the concept of “Career Planning”. Percentage distributions were used for categorical variables and mean ± standard deviation calculations were used for numerical variables. Student’s t-test was used to compare students’ characteristics with career planning and important factors, and Pearson correlation test was used to evaluate competency areas with the mean scores of important factors in career planning. Statistical significance level was accepted as p<0.05. Thematic analysis was applied to qualitative data. Results The perception of the term “career planning” revealed 12 themes of which “life” was the most common (140/245; 57.1%), followed by “professional life” (102/245; 41.6%). Participants’ total satisfaction rate was found 65.6% (Mean=39.36 ± 14.88) for the course. Students were most satisfied with the goals and content appropriateness (7.77 ± 3.08). The stimulating and motivating attribute of the CPC was the least satisfactory item. Conclusions An evaluation report, based on students’ feedback, was shared with the faculty members involved in education via an interactive web page. The survey not only benefitted course educators but also helped students to reflect on the course content. CPC can help students to address their strengths and weaknesses and hopefully to take supportive initiatives at the beginning of their career.