Search results for: clinical markers

Authors: Leyla Sattarzadeh, Shahin Fathi Molk Kian, Maghsoud Peeri, Mohammadali Azarbaijani, Hasan Matin Homaee

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Inflammation by various mechanisms may cause atherosclerosis. Systemic circulating inflammatory markers such as C-reactive protein (CRP), pro-inflammatory cytokines such as Interleukin-6 (IL-6), vascular inflammatory markers as adhesion molecules like Intracellular Adhesion Molecule-1 (ICAM-1) and Vascular Cell Adhesion Molecule-1 (VCAM-1) are the predictors of cardiovascular diseases. Regarding the conflicting results about the effect of different patterns of resistance exercise training on these inflammatory markers, present study aimed to examine the effect of different patterns of eight week resistance exercise training on CRP, IL-6, ICAM-1 and VCAM-1 levels in healthy untrained women. 56 healthy volunteered untrained female university students (aged: 21 ± 3 yr., Body Mass Index: 21.5 ± 3.5 kg/m²) were selected purposefully and divided into four groups. At the end of training protocol and after subject drop during the protocol, upper body exercise training (n=11), lower body (n=12) and whole body resistance exercise training group (n=11) completed the eight weeks of training period although the control group (n=7) did anything. Blood samples gathered pre and post-experimental period and CRP, IL-6, ICAM-1 and VCAM-1 levels were evaluated using special laboratory kits, then the difference of pre and post values of each indices analyzed using one-way analysis of variance (α < 0.05). The results of one way ANOVA for difference of pre and post values of CRP, ICAM-1 and VCAM-1 showed no significant changes due to the exercise training, but there were significant differences between groups about IL-6. Tukey post- hoc test indicated that there is significant difference between the differences of pre and post values of IL-6 between lower body exercise training group and control group, and eight weeks of lower body exercise training lead to significant changes in IL-6 values. There were no changes in anthropometric indices. The findings show that the different patterns of upper, lower and whole body exercise training by involving the different amounts of muscles altered the IL-6 values in lower body exercise training group probably because of engaging the bigger amount of muscles, but showed any significant changes about CRP, ICAM-1 and VCAM-1 probably due to intensity and duration of exercise or the lower levels of these markers at baseline of healthy people.

Keywords: resistance training, C-reactive protein, interleukin-6, intracellular adhesion molecule-1, vascular cell adhesion molecule-1

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Authors: Fasil Wagnew, Kefyalew Addis Alene, Setegn Eshetie, Tom Wingfield, Matthew Kelly, Darren Gray

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Introduction: Undernutrition is a major and under-appreciated risk factor for TB, which is estimated to be responsible for 1.9 million TB cases per year globally. The effectiveness of micronutrient supplementation on TB treatment outcomes and its prognostic markers such as sputum conversion and serum zinc, retinol, and hemoglobin levels has been poorly understood. This systematic review and meta-analysis aimed to determine the association between zinc and vitamin A supplementation and TB treatment outcomes and its prognostic markers. Methods: A systematic literature search for randomized controlled trials (RCTs) was performed in PubMed, Embase, and Scopus databases. Meta-analysis with a random effect model was performed to estimate risk ratio (RR) and mean difference (MD), with a 95% confidence interval (CI), for dichotomous and continuous outcomes, respectively. Results: Our search identified 2,195 records. Of these, nine RCTs consisting of 1,375 participants were included in the final analyses. Among adults with pulmonary TB, zinc (RR: 0.94, 95%CI: 0.86, 1.03), vitamin A (RR: 0.90, 95%CI: 0.80, 1.01), and combined zinc and vitamin A (RR: 0.98, 95%CI: 0.89, 1.08) supplementation were not significantly associated with TB treatment success. Combined zinc and vitamin A supplementation was significantly associated with increased sputum smear conversion at 2 months (RR: 1.16, 95%CI: 1.03, 1.32), serum zinc levels at 2 months (MD of 0.86umol/l, 95% CI: 0.14, 1.57), serum retinol levels at 2 months (MD: 0.06umol/l, 95 % CI: 0.04, 0.08) and 6 months (MD: 0.12umol/l, 95 % CI: 0.10, 0.14), and serum hemoglobin level at 6 months (MD: 0.29 ug/dl, 95% CI: 0.08 to 0.51), among adults with TB. Conclusions: Providing zinc and vitamin A supplementation to adults with pulmonary TB during treatment may increase early sputum smear conversion, serum zinc, retinol, and hemoglobin levels. However, the use of zinc, vitamin A, or both were not associated with TB treatment success.

Keywords: zinc and vitamin A supplementation, tuberculosis, treatment outcomes, meta-analysis, RCT

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Authors: Sohyoung Won, Heebal Kim, Dajeong Lim

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Genomic prediction is an effective way to measure the abilities of livestock for breeding based on genomic estimated breeding values, statistically predicted values from genotype data using best linear unbiased prediction (BLUP). Using haplotypes, clusters of linked single nucleotide polymorphisms (SNPs), as markers instead of individual SNPs can improve the reliability of genomic prediction since the probability of a quantitative trait loci to be in strong linkage disequilibrium (LD) with markers is higher. To efficiently use haplotypes in genomic prediction, finding optimal ways to define haplotypes is needed. In this study, 770K SNP chip data was collected from Hanwoo (Korean cattle) population consisted of 2506 cattle. Haplotypes were first defined in three different ways using 770K SNP chip data: haplotypes were defined based on 1) length of haplotypes (bp), 2) the number of SNPs, and 3) k-medoids clustering by LD. To compare the methods in parallel, haplotypes defined by all methods were set to have comparable sizes; in each method, haplotypes defined to have an average number of 5, 10, 20 or 50 SNPs were tested respectively. A modified GBLUP method using haplotype alleles as predictor variables was implemented for testing the prediction reliability of each haplotype set. Also, conventional genomic BLUP (GBLUP) method, which uses individual SNPs were tested to evaluate the performance of the haplotype sets on genomic prediction. Carcass weight was used as the phenotype for testing. As a result, using haplotypes defined by all three methods showed increased reliability compared to conventional GBLUP. There were not many differences in the reliability between different haplotype defining methods. The reliability of genomic prediction was highest when the average number of SNPs per haplotype was 20 in all three methods, implying that haplotypes including around 20 SNPs can be optimal to use as markers for genomic prediction. When the number of alleles generated by each haplotype defining methods was compared, clustering by LD generated the least number of alleles. Using haplotype alleles for genomic prediction showed better performance, suggesting improved accuracy in genomic selection. The number of predictor variables was decreased when the LD-based method was used while all three haplotype defining methods showed similar performances. This suggests that defining haplotypes based on LD can reduce computational costs and allows efficient prediction. Finding optimal ways to define haplotypes and using the haplotype alleles as markers can provide improved performance and efficiency in genomic prediction.

Keywords: best linear unbiased predictor, genomic prediction, haplotype, linkage disequilibrium

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Authors: Thao Nguyen, Maximiliano Hyon, Sany Rajagukguk, Anna Melkonyan

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Introduction: Type 2 Diabetes is a well-established risk factor for severe SARS-CoV-2 infection. Uncontrolled hyperglycemia in patients with established or newly diagnosed diabetes is associated with poor outcomes, including increased mortality and hospital length of stay. Objectives: Our study aims to compare three different glycemic management strategies and their association with clinical outcomes in patients hospitalized for moderate to severe SARS-CoV-2 infection. Identifying optimal glycemic management strategies will improve the quality of patient care and improve their outcomes. Method: This is a retrospective observational study on patients hospitalized at Adventist Health White Memorial with severe SARS-CoV-2 infection from 11/1/2020 to 02/28/2021. The following inclusion criteria were used: positive SARS-CoV-2 PCR test, age >18 yrs old, diabetes or random glucose >200 mg/dL on admission, oxygen requirement >4L/min, and treatment with glucocorticoids. Our exclusion criteria included: ICU admission within 24 hours, discharge within five days, death within five days, and pregnancy. The patients were divided into three glycemic management groups: Group 1, managed solely by the Primary Team, Group 2, by Pharmacy; and Group 3, by Endocrinologist. Primary outcomes were average glucose on Day 5, change in glucose between Days 3 and 5, and average insulin dose on Day 5 among groups. Secondary outcomes would be upgraded to ICU, inpatient mortality, and hospital length of stay. For statistics, we used IBM® SPSS, version 28, 2022. Results: Most studied patients were Hispanic, older than 60, and obese (BMI >30). It was the first CV-19 surge with the Delta variant in an unvaccinated population. Mortality was markedly high (> 40%) with longer LOS (> 13 days) and a high ICU transfer rate (18%). Most patients had markedly elevated inflammatory markers (CRP, Ferritin, and D-Dimer). These, in combination with glucocorticoids, resulted in severe hyperglycemia that was difficult to control. Average glucose on Day 5 was not significantly different between groups primary vs. pharmacy vs. endocrine (220.5 ± 63.4 vs. 240.9 ± 71.1 vs. 208.6 ± 61.7 ; P = 0.105). Change in glucose from days 3 to 5 was not significantly different between groups but trended towards favoring the endocrinologist group (-26.6±73.6 vs. 3.8±69.5 vs. -32.2±84.1; P= 0.052). TDD insulin was not significantly different between groups but trended towards higher TDD for the endocrinologist group (34.6 ± 26.1 vs. 35.2 ± 26.4 vs. 50.5 ± 50.9; P=0.054). The endocrinologist group used significantly more preprandial insulin compared to other groups (91.7% vs. 39.1% vs. 65.9% ; P < 0.001). The pharmacy used more basal insulin than other groups (95.1% vs. 79.5% vs. 79.2; P = 0.047). There were no differences among groups in the clinical outcomes: LOS, ICU upgrade, or mortality. Multivariate regression analysis controlled for age, sex, BMI, HbA1c level, renal function, liver function, CRP, d-dimer, and ferritin showed no difference in outcomes among groups. Conclusion: Given high-risk factors in our population, despite efforts from the glycemic management teams, it’s unsurprising no differences in clinical outcomes in mortality and length of stay.

Keywords: glycemic management, strategies, hospitalized, SARS-CoV-2, outcomes

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Authors: Huma Balouch

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Menochilus sexmaculatus commonly known as six spotted zig zag ladybird, is an aphidophagus and the most misidentified Coccinellids due to the occurrence of numerous color variants. The correct identification of Menochilus sexmaculatus and its strains is necessary to implement the use of biological control. In the present study phenotypic and genotypic polymorphism was investigated in Menochilus sexmaculatus collected from Punjab, NWFP and Sindh provinces of Pakistan. Six different morphs of the species were distinguished by analyzing its Elytral color and spot pattern and then Polymerase Chain Reaction was used to generate random amplification of polymorphic DNA (RAPD) from six different types of Menochilus sexmaculatus. Forty primers (OPA & OPC Kit) were used to perform RAPD PCR on six different types of Menochilus sexmaculatus of which, seven primers revealed different patterns related to the Menochilus sexmaculatus types. These seven primers (OPA-04, OPA-09, OPA-18, OPC-04, OPC-12, OPC-15 and OPC-18) produced 111 clear polymorphic bands and 6 scorable strain specific markers. The cluster analysis applied to RAPD data showed high polymorphism among six types and it can be concluded that these six types are six polymorphic strains of the same species.

Keywords: Menochilus sexmaculatus, aphidophagus, coccinellids, phenotypic and genotypic polymorphism, RAPD-PCR, strain specific markers

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Authors: Hossein Ghaedamini, Salman Farahbakhsh, Alireza Amirbeigi, Zahra Saghafi, Salman Daneshi, Alireza Ghaedamini

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Background: Assessing the challenges of clinical education of medical students is one of the most important and sensitive parts of medical education. The aim of this study was to investigate the views and experiences of Kerman medical students on the factors that facilitate and inhibit the quality of clinical education. Materials and Methods: This research was qualitative and used a phenomenological approach. The study population included medical interns of Kerman University of Medical Sciences in 1400. The method of data collection was in-depth interviews with participants. Data were encoded and analyzed by Claizey stepwise model. Results: First, about 540 primary codes were extracted in the form of two main themes (facilitators and inhibitors) and 10 sub-themes including providing motivational models and creating interest in interns, high scientific level of professors and the appropriate quality of their teaching, the use of technology in the clinical education process, delegating authority and freedom of action and more responsibilities to interns, inappropriate treatment of some officials, professors, assistants and department staff with their interns, inadequate educational programming, lack of necessary cooperation and providing inappropriate treatment by clinical training experts for interns, inadequate evaluation method in clinical training for interns, poor quality mornings, the unefficiency of grand rounds, the inappropriate way of evaluating clinical training for interns, the lack of suitable facilities and conditions with the position of a medical intern, and the hardwork of some departments were categorized. Conclusion: Clinical education is always mixed with special principles and subtleties, and special attention to facilitators and inhibitors in this process has an important role in improving its quality.

Keywords: clinical education, medical students, qualitative study, education

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Authors: Lu-Chiu Huang, Pei-Pei Chen, Li-Chin Yu, Chiao-Wen Kuo, Tsui-Tao Liu, Rung-Chuang Feng

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Purpose: People put increasing emphasis on demands of medical quality and protecting their interests. Patients' or family's dissatisfaction with medical care may easily lead to medical dispute. Physician-family communication plays an essential role in medical care. A sound communication cannot only strengthen patients' belief in the medical team but make patient have definite insight into treatment course of the disease. A family meeting provides an effective platform for communication between clinical staff, patients and family. Decisions and consensuses formed in family meetings can promote patients' or family's satisfaction with medical care. Clinical staff's attitudes toward family meeting may determine behavioral intentions to hold family meeting. This study aims to explore clinical staff's difficulties in holding family meeting and evaluate how their attitudes and behavior influence the effect of family meetings. Methods: This was a cross-sectional study. It was conducted at a regional teaching hospital in Taipei city. The research team developed its own structural questionnaires, whose expert validity was checked by the nursing experts. Participants filled in the questionnaires online. Data were collected by convenience sampling. A total of 568 participants were invited. They included doctors, nurses, social workers, and so on. Results: 1) The average score of ‘clinical staff’s attitudes to family meetings’ was 5.15 (SD=0.898). It fell between ‘somewhat agree’ and ‘mostly agree’ on the 7-point likert scale. It indicated that clinical staff had positive attitudes toward family meetings, 2) The average score of ‘clinical staff’s behavior to family meetings’ was 5.61 (SD=0.937). It fell between ‘somewhat agree’ and ‘mostly agree’ on the 7-point likert scale. It meant clinical staff tended to have positive behavior at the family meeting, and 3) The average score of ‘Difficulty in conducting family meetings’ was 5.15 (SD=0.897). It fell between ‘somewhat agree’ and ‘mostly agree’ on the 7-point likert scale. The higher the score was, the less difficulty the clinical staff felt. It demonstrated clinical staff felt less difficulty in conducting family meetings. Clinical staff's identification with family meetings brought favored effects. Persistent and active promotion for family meetings can bring patients and family more benefits. Implications for practice: Understanding clinical staff's difficulty in participating family meeting and exploring their attitudes or behavior toward physician-family communication are helpful to develop modes of interaction. Consequently, quality and satisfaction of physician-family communication can be increased.

Keywords: clinical staff, communication, family meeting, physician-family

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Authors: Phawida Wattanasoonthorn

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Ventilator-associated pneumonia is a serious infection found to be among the top three infections in the hospital. To investigate the effects of clinical practice guideline on knowledge and preventive practices of nursing personnel, and incidences of ventilator-associated pneumonia. A pre-post quasi-experimental study on 17 professional nurses, and 123 ventilator-associated pneumonia patients admitted to the surgical intensive care unit, and the accident and surgical ward of Songkhla Hospital from October 2013 to January 2014. The study found that after using the clinical practice guideline, the subjects’ median score increased from 16.00 to 19.00. The increase in practicing correctly was from 66.01 percent to 79.03 percent with the statistical significance level of .05, and the incidences of ventilator-associated pneumonia decreased by 5.00 percent. The results of this study revealed that the use of the clinical practice guideline helped increase knowledge and practice skill of nursing personnel, and decrease incidences of ventilator-associated pneumonia. Thus, nursing personnel should be encouraged, reminded and promoted to continue using the practice guideline through various means including training, providing knowledge, giving feedback, and putting up posters to remind them of practicing correctly and sustainably.

Keywords: Clinical Practice Guideline, knowledge, Preventive Ventilator, Pneumonia

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Authors: Valeriy M. Kondratev, Ekaterina A. Vyacheslavova, Talgat Shugabaev, Alexander S. Gudovskikh, Alexey D. Bolshakov

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Modern sensorics imposes strict requirements on the biosensors characteristics, especially technological feasibility, and selectivity. There is a growing interest in the analysis of human health biological markers, which indirectly testifying the pathological processes in the body. Such markers are acids and alkalis produced by the human, in particular - ammonia and hydrochloric acid, which are found in human sweat, blood, and urine, as well as in gastric juice. Biosensors based on modern nanomaterials, especially low dimensional, can be used for this markers detection. Most classical adsorption sensors based on metal and silicon oxides are considered non-selective, because they identically change their electrical resistance (or impedance) under the action of adsorption of different target analytes. This work demonstrates a feasible frequency-resistive method of electrical impedance spectroscopy data analysis. The approach allows to obtain of selectivity in adsorption sensors of a resistive type. The method potential is demonstrated with analyzis of impedance spectra of silicon nanowires in the presence of NH3 and HCl vapors with concentrations of about 125 mmol/L (2 ppm) and water vapor. We demonstrate the possibility of unambiguous distinction of the sensory signal from NH3 and HCl adsorption. Moreover, the method is found applicable for analysis of the composition of ammonia and hydrochloric acid vapors mixture without water cross-sensitivity. Presented silicon sensor can be used to find diseases of the gastrointestinal tract by the qualitative and quantitative detection of ammonia and hydrochloric acid content in biological samples. The method of data analysis can be directly translated to other nanomaterials to analyze their applicability in the field of biosensory.

Keywords: electrical impedance spectroscopy, spectroscopy data analysis, selective adsorption sensor, nanotechnology

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Authors: G. Cunningham, E. Cantor, X. Wu, F. Shen, G. Jiang, S. Philips, C. Bales, Y. Xiao, T. R. Cummins, J. C. Fehrenbacher, B. P. Schneider

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Background: Taxane-induced peripheral neuropathy (TIPN) is the most devastating survivorship issue for patients receiving therapy. Dose reductions due to TIPN in the curative setting lead to inferior outcomes for African American patients, as prior research has shown that this group is more susceptible to developing severe neuropathy. The mechanistic underpinnings of TIPN, however, have not been entirely elucidated. While it would be appealing to use primary tissue to study the development of TIPN, procuring nerves from patients is not realistically feasible, as nerve biopsies are painful and may result in permanent damage. Therefore, our laboratory has investigated paclitaxel-induced neuronal morphological and molecular changes using an ex vivo model of human-induced pluripotent stem cell (iPSC)-derived neurons. Methods: iPSCs are undifferentiated and endlessly dividing cells that can be generated from a patient’s somatic cells, such as peripheral blood mononuclear cells (PBMCs). We successfully reprogrammed PBMCs into iPSCs using the Erythroid Progenitor Reprograming Kit (STEMCell Technologiesᵀᴹ); pluripotency was verified by flow cytometry analysis. iPSCs were then induced into neurons using a differentiation protocol that bypasses the neural progenitor stage and uses selected small-molecule modulators of key signaling pathways (SMAD, Notch, FGFR1 inhibition, and Wnt activation). Results: Flow cytometry analysis revealed expression of core pluripotency transcription factors Nanog, Oct3/4 and Sox2 in iPSCs overlaps with commercially purchased pluripotent cell line UCSD064i-20-2. Trilineage differentiation of iPSCs was confirmed with immunofluorescent imaging with germ-layer-specific markers; Sox17 and ExoA2 for ectoderm, Nestin, and Pax6 for mesoderm, and Ncam and Brachyury for endoderm. Sensory neuron markers, β-III tubulin, and Peripherin were applied to stain the cells for the maturity of iPSC-derived neurons. Patch-clamp electrophysiology and calcitonin gene-related peptide (CGRP) release data supported the functionality of the induced neurons and provided insight into the timing for which downstream assays could be performed (week 4 post-induction). We have also performed a cell viability assay and fluorescence-activated cell sorting (FACS) using four cell-surface markers (CD184, CD44, CD15, and CD24) to select a neuronal population. At least 70% of the cells were viable in the isolated neuron population. Conclusion: We have found that these iPSC-derived neurons recapitulate mature neuronal phenotypes and demonstrate functionality. Thus, this represents a patient-derived ex vivo neuronal model to investigate the molecular mechanisms of clinical TIPN.

Keywords: chemotherapy, iPSC-derived neurons, peripheral neuropathy, taxane, paclitaxel

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Authors: Yiding Qu, Svetlana V. Komarova

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Chemokines acting through their cognate chemokine receptors guide the directional migration of the cell along the chemokine gradient. Several chemokine receptors were recently identified as non-signaling (decoy), based on their ability to bind the chemokine but produce no measurable signal in the cell. The function of these decoy receptors is not well understood. We examined the expression of a decoy receptor CCRL1 and a signaling receptor that binds to the same ligands, CCR7, in prostate cancer using publically available microarray data (www.oncomine.org). The expression of both CCRL1 and CCR7 increased in an approximately half of prostate carcinoma samples and the majority of metastatic cancer samples compared to normal prostate. Moreover, the expression of CCRL1 positively correlated with the expression of CCR7. These data suggest that CCR7 and CCRL1 can be used as clinical markers for the early detection of transformation from carcinoma to metastatic cancer. In addition, these data support our hypothesis that the non-signaling chemokine receptors actively stimulate cell migration.

Keywords: bioinformatics, cell migration, decoy receptor, meta-analysis, prostate cancer

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Authors: Getachew Belay Kassahun

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Background: Locally established clinical laboratory reference intervals (RIs) are required to interpret laboratory test results for screening, diagnosis, and prognosis. The objective of this study was to establish a reference interval of clinical chemistry parameters among apparently healthy adolescents aged between 12 and 17 years in Mekelle, Tigrai, in the northern part of Ethiopia. Methods: Community-based cross-sectional study was employed from December 2018 to March 2019 in Mekelle City among 172 males and 172 females based on a Multi-stage sampling technique. Blood samples were tested for Fasting blood sugar (FBS), alanine amino transferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), Creatinine, urea, total protein, albumin (ALB), direct and indirect bilirubin (BIL.D and BIL.T) using 25 Bio system clinical chemistry analyzer. Results were analyzed using SPSS version 23 software and based on the Clinical Laboratory Standard Institute (CLSI)/ International Federation of Clinical Chemistry (IFCC) C 28-A3 Guideline which defines the reference interval as the 95% central range of 2.5th and 97.5th percentiles. Mann Whitney U test, descriptive statistics and box and whisker were statistical tools used for analysis. Results: This study observed statistically significant differences between males and females in ALP, ALT, AST, Urea and Creatinine Reference intervals. The established reference intervals for males and females, respectively, were: ALP (U/L) 79.48-492.12 versus 63.56-253.34, ALT (U/L) 4.54-23.69 versus 5.1-20.03, AST 15.7- 39.1 versus 13.3- 28.5, Urea (mg/dL) 9.33-24.99 versus 7.43-23.11, and Creatinine (mg/dL) 0.393-0.957 versus 0.301-0.846. The combined RIs for Total Protein (g/dL) were 6.08-7.85, ALB (g/dL) 4.42-5.46, FBS(mg/dL) 65-110, BIL.D (mg/dL) 0.033-0.532, and BIL.T (mg/dL) 0.106-0.812. Conclusions: The result showed a marked difference between sex and company-derived values for selected clinical chemistry parameters. Thus, the use of age and sex-specific locally established reference intervals for clinical chemistry parameters is recommended.

Keywords: reference interval, adolescent, clinical chemistry, Ethiopia

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Authors: Daphne Alroy-Thiberge

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Trauma-impacted individuals present unique treatment challenges that include high reactivity, hyper-and hypo-arousal, poor adherence to therapy, as well as powerful transference and counter-transference experiences in therapy. This work provides an overview of the clinical tenets most often encountered in trauma-impacted individuals. Further, it provides readily applicable clinical techniques to optimize therapeutic rapport and facilitate accelerated positive mental health outcomes. Finally, integrated neuroscience and clinical evidence-based data are discussed to shed new light on crisis states in trauma-impacted individuals. This knowledge is utilized to provide effective and concrete interventions towards rapid and successful de-escalation of the impacted individual. A highly interactive, adult-learning-principles-based modality is utilized to provide an organic learning experience for participants. The information and techniques learned aim to increase clinical effectiveness, reduce staff injuries and burnout, and significantly enhance positive mental health outcomes and self-determination for the trauma-impacted individuals treated.

Keywords: clinical competencies, crisis interventions, psychotherapy techniques, trauma informed care

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Authors: Zahra F. Al-Khateeb, Shenel Shekerzade, Hasna Boumenar, Siân M. Henson, Jordi L. Tremoleda, A. T. Michael-Titus

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Traumatic brain injury (TBI) is the leading cause of disability and death in young adults and also increases the risk ofneurodegeneration. The mechanisms linking moderate to severe TBI to neurodegeneration are not known. It has been proposed that cellular senescence inductionpost-injury could amplify neuroinflammation and induce long-term changes. The impact of these processes after injury to an immature brain has not been characterised yet. We carried out a controlled cortical impact injury (CCI) in juvenile 1 month-old male CD1 mice. Animals were anesthetised and received a unilateral CCI injury. The sham group received anaesthesia and had a craniotomy. A naïve group had no intervention. The brain tissue was analysed at 5 days and 35 days post-injury using immunohistochemistry and markers for microglia, astrocytes, and senescence. Compared tonaïve animals, injured mice showed an increased microglial and astrocytic reaction early post-injury, as reflected in Iba1 and GFAP markers, respectively; the GFAP increase persisted in the later phase. The senescence analysis showed a significant increase inγH2AX-53BP1 nuclear foci, 8-oxoguanine, p19ARF, p16INK4a, and p53 expression in naïve vs. sham groups and naïve vs. CCI groups, at 5 dpi. At 35 days, the difference was no longer statistically significant in all markers. The injury induced a decrease p21 expression vs. the naïve group, at 35 dpi. These results indicate the induction of a complex senescence response after immature brain injury. Some changes occur early and may reflect the activation/proliferation of non-neuronal cells post-injury that had been hindered, whereas changes such as p21 downregulation may reflect a delayed response and pro-repair processes.

Keywords: cellular senescence, traumatic brain injury, brain injury, controlled cortical impact

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Authors: Alexey Yu. Martynov, Sulejman Bayramov

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Introduction: The myocardial bridging is the most common anomaly of the coronary arteries (CA). Depending on the examination method, the frequency of detected myocardial bridges (MB) varies in a rather wide range. The typical clinical manifestations of MB are angina pectoris, arrhythmias, sudden cardiac death. Objective: To study the incidence of MB in patients hospitalized with coronary artery disease (CAD). To assess clinical manifestations of MB in patients admitted with CAD. Materials and methods: A retrospective analysis of 19159 case histories of patients admitted at clinical city hospital in Moscow from 01.01.2018 to 31.12 2019 with CAD was performed. 9384 patients’ coronary angiographies (CAG) were examined for MB. The localization of MB, the degree of coronary contraction by MB, the number of MB, isolated MB and combined with CAD were assessed. The clinical manifestations of MB were determined. Results: MB was detected in 52 patients all with one myocardial bridge. 20 patients with MB have intact CA, and 32 patients have MB combined with CAD. Among 20 patients with intact CA: I degree of MB contraction (up to 50%) was detected in 9 patients. Clinical manifestations in five cases were angina pectoris, in 3 myocardial infarction (MI) - 1 patients with ST segment elevation MI (STEMI), 2 without ST segment elevation MI (NSTEMI), 1 post-infarction cardiosclerosis (PICS). Stable angina II FC in 3, III FC in 1, vasospastic angina (VSA) in 1 patient. II degree of MB contraction (up to 50-70%) was determined in 9 patients: in seven cases angina pectoris was detected, 1 NSTEMI, 1 PICS. Stable angina II FC in 3, III FC in 1, VSA in 3 patients. III degree of MB contraction (> 70%) detected in 2 patients. II FC stable angina in one case, PICS in another. Among 32 patients having MB combined with CAD I degree of MB contraction was observed in 20 patients. Clinical manifestations in 12 cases were angina pectoris in 8 II FC and in 4 III FC, 7 MI 6 with STEMI and 1 NSTEMI, 1 PICS. II degree of MB contraction was detected in 7 patients, 4 of them had angina pectoris, 3 MI 2 with STEMI and 1 NSTEMI. Stable angina II FC in 3, VSA in 1 patients. III degree of MB contraction was diagnosed in five patients. In two cases, II FC and III FC stable angina were observed, 2 MI with STEMI and NSTEMI, 1 PICS. Conclusions: MB incidence is one in 368 patients with CAD. The most common involvement (68%) is MB combined with CA atherosclerotic lesions. MB with intact CA are detected in one-third (32%) of patients. The first-degree MB contraction is most frequent condition. MI is more often detected in intact CA with first degree MB than in the second degree. The degree of MB contraction was not correlated with the severity of the clinical manifestations.

Keywords: clinical manifestations, coronary angiography, coronary artery disease, myocardial bridging, myocardial infarction, stable angina

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Authors: Kamal Kaushik, Trisha, Dandapni, Sambit Nanda, A. Mukherjee, S. Pradhan

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INTRODUCTION: As complexity in radiotherapy practice and processes are increasing, there is a need to assure quality control to a greater extent. At present, no international literature available with regards to the optimal quality control indicators for radiotherapy; moreover, few clinical audits have been conducted in the field of radiotherapy. The primary aim is to improve the processes that directly impact clinical outcomes for patients in terms of patient safety and quality of care. PROCEDURE: A team of an Oncologist, a Medical Physicist and a Radiation Therapist was formed for weekly clinical audits of patient’s undergoing radiotherapy audits The stages for audits include Pre planning audits, Simulation, Planning, Daily QA, Implementation and Execution (with image guidance). Errors in all the parts of the chain were evaluated and recorded for the development of further departmental protocols for radiotherapy. EVALUATION: The errors at various stages of radiotherapy chain were evaluated and recorded for comparison before starting the clinical audits in the department of radiotherapy and after starting the audits. It was also evaluated to find the stage in which maximum errors were recorded. The clinical audits were used to structure standard protocols (in the form of checklist) in department of Radiotherapy, which may lead to further reduce the occurrences of clinical errors in the chain of radiotherapy. RESULTS: The aim of this study is to perform a comparison between number of errors in different part of RT chain in two groups (A- Before Audit and B-After Audit). Group A: 94 pts. (48 males,46 female), Total no. of errors in RT chain:19 (9 needed Resimulation) Group B: 94 pts. (61 males,33 females), Total no. of errors in RT chain: 8 (4 needed Resimulation) CONCLUSION: After systematic periodic clinical audits percentage of error in radiotherapy process reduced more than 50% within 2 months. There is a great need in improving quality control in radiotherapy, and the role of clinical audits can only grow. Although clinical audits are time-consuming and complex undertakings, the potential benefits in terms of identifying and rectifying errors in quality control procedures are potentially enormous. Radiotherapy being a chain of various process. There is always a probability of occurrence of error in any part of the chain which may further propagate in the chain till execution of treatment. Structuring departmental protocols and policies helps in reducing, if not completely eradicating occurrence of such incidents.

Keywords: audit, clinical, radiotherapy, improving functionality

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Authors: Yousif Mohamed Y. Abdallah, Mommen A. Alkhir, Amel S. Algaddal

Abstract:

This was experimental study conducted to study segmentation of brain in MRI images using edge detection and morphology filters. For brain MRI images each film scanned using digitizer scanner then treated by using image processing program (MatLab), where the segmentation was studied. The scanned image was saved in a TIFF file format to preserve the quality of the image. Brain tissue can be easily detected in MRI image if the object has sufficient contrast from the background. We use edge detection and basic morphology tools to detect a brain. The segmentation of MRI images steps using detection and morphology filters were image reading, detection entire brain, dilation of the image, filling interior gaps inside the image, removal connected objects on borders and smoothen the object (brain). The results of this study were that it showed an alternate method for displaying the segmented object would be to place an outline around the segmented brain. Those filters approaches can help in removal of unwanted background information and increase diagnostic information of Brain MRI.

Keywords: improvement, brain, matlab, markers, boundaries

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Authors: S. A. Bhat, Ahmad Arif, Muneeb U. Rehman, Manzoor R Mir, S. Bilal, Ishraq Hussain, H. M Khan, S. Shanaz, M. I Mir, Sabhiya Majid

Abstract:

Background: Geologically Climatic conditions of the state range from sub-tropical (Jammu), temperate (Kashmir) to cold artic (Ladakh) zones, which exerts significant influence on its agro-climatic conditions. Gastrointestinal parasitism is a major problem in sheep production worldwide. Materials and Methods: The present study was to evaluate the resistance status of sheep breeds reared in Kashmir Valley for natural resistance against Haemonchus contortus by natural pasture challenge infection. Ten microsatellite markers were used in the study for evaluation of association of Ovar-MHC with parasitic resistance in association with biochemical and parasitological parameters. Following deworming, 500 animals were subjected to selected contaminated pastures in a vicinity of the livestock farms of SKUAST-K and Sheep Husbandry Kashmir. For each animal about 10-15 ml blood was collected aseptically for molecular and biochemical analysis. Weekly fecal samples (3g) were taken, directly from the rectum of all experimental animals and examined for Fecal egg count (FEC) with modified McMaster technique. Packed cell volume (PCV) was determined within 2-5 h of blood collection, all the biochemical parameters were determined in serum by semi automated analyzer. DNA was extracted from all the blood samples with phenol-chloroform method. Microsatellite analysis was done by denaturing sequencing gel electrophoresis Results: Overall sheep from Bakerwal breed followed by Corriediale breed performed relatively better in the trial; however difference between breeds remained low. Both significant (P<0.05) and non-significant differences with respect to resistance against haemonchosis were noted at different intervals in all the parameters.. All the animals were typed for the microsatellites INRA132, OarCP73, DRB1 (U0022), OLA-DQA2, BM1818, TFAP2A, HH56, BM1815, IL-3 and BM-1258. An association study including the effect of FEC, PCV, TSP, SA, LW, and the number of alleles within each marker was done. All microsatellite markers showed degree of heterozygosity of 0.72, 0.72, 0.75, 0.62, 0.84, 0.69, 0.66, 0.65, 0.73 and 0.68 respectively. Significant association between alleles and the parameters measured were only found for the OarCP73, OLA-DQA2 and BM1815 microsatellite marker. Standard alleles of the above markers showed significant effect on the TP, SA and body weight. The three sheep breeds included in the study responded differently to the nematode infection, which may be attributed to their differences in their natural resistance against nematodes. Conclusion: Our data confirms that some markers (OarCP73, OLA-DQA2 and BM1815) within Ovar-MHC are associated with phenotypic parameters of resistance and suggest superiority of Bakerwal sheep breed in natural resistance against Haemonchus contortus.

Keywords: Ovar-Mhc, ovine leukocyte antigen (OLA), sheep, parasitic resistance, Haemonchus contortus, phenotypic & genotypic markers

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Authors: Sandeep Kumar Agrawal, Aditi Bhattacharya, Janvie Manhas, Krushna Bhatt, Yatin Kholakiya, Nupur Khera, Ajoy Roychoudhury, Sudip Sen

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Autologous cultured explants of human oral mucosal epithelial cells (OMEC) are a potential therapeutic modality for limbal stem cell deficiency (LSCD). Injury or inflammation of the ocular surface in the form of burns, chemicals, Stevens Johnson syndrome, ocular cicatricial pemphigoid etc. can lead to destruction and deficiency of limbal stem cells. LSCD manifests in the form of severe ocular surface diseases (OSD) characterized by persistent and recurrent epithelial defects, conjuntivalisation and neovascularisation of the corneal surface, scarring and ultimately opacity and blindness. Most of the cases of OSD are associated with severe dry eye pertaining to diminished mucin and aqueous secretion. Mycophenolate mofetil (MMF) has been shown to upregulate the mucin expression in conjunctival goblet cells in vitro. The aim of this study was to evaluate the effects of MMF on mucin expression in primary cultures of oral mucosal epithelial cells. With institutional ethics committee approval and written informed consent, thirty oral mucosal epithelial tissue samples were obtained from patients undergoing oral surgery for non-malignant conditions. OMEC were grown on human amniotic membrane (HAM, obtained from expecting mothers undergoing elective caesarean section) scaffold for 2 weeks in growth media containing DMEM & Ham’s F12 (1:1) with 10% FBS and growth factors. In vitro dosage of MMF was standardised by MTT assay. Analysis of stem cell markers was done using RT-PCR while mucin mRNA expression was quantified using RT-PCR and q-PCR before and after treating cultured OMEC with graded concentrations of MMF for 24 hours. Protein expression was validated using immunocytochemistry. Morphological studies revealed a confluent sheet of proliferating, stratified oral mucosal epithelial cells growing over the surface of HAM scaffold. The presence of progenitor stem cell markers (p63, p75, β1-Integrin and ABCG2) and cell surface associated mucins (MUC1, MUC15 and MUC16) were elucidated by RT-PCR. The mucin mRNA expression was found to be upregulated in MMF treated primary cultures of OMEC, compared to untreated controls as quantified by q-PCR with β-actin as internal reference gene. Increased MUC1 protein expression was validated by immunocytochemistry on representative samples. Our findings conclude that OMEC have the ability to form a multi-layered confluent sheet on the surface of HAM similar to a cornea, which is important for the reconstruction of the damaged ocular surface. Cultured OMEC has stem cell properties as demonstrated by stem cell markers. MMF can be a novel enhancer of mucin production in OMEC. It has the potential to improve dry eye in patients undergoing OMEC transplantation for bilateral OSD. Further clinical trials are required to establish the role of MMF in patients undergoing OMEC transplantation.

Keywords: limbal stem cell deficiency, mycophenolate mofetil, mucin, ocular surface disease

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Authors: Anoosh Eghdami

Abstract:

Background and aim: Conventional anticancer drugs display significant shortcomings which limit their use in cancer therapy. For this reason, important progress has been achieved in the field of nanotechnology to solve these problems and offer a promising and effective alternative for cancer treatment. Tumor markers may also be measured periodically during cancer therapy. Tumor markers may also be measured after treatment has ended to check for recurrence the return of cancer. The aim of this study was to evaluate the effect of nano drug delivery in induced breast cancer with DMBA by using CA15-3 tumor marker. Material and method: the rats were divided into five groups. The first group (control n=15) were fed only sesame oil as a gavage. In the second group n=15,10 mg DMBA was dissolved in 5ml of sesame oil and were fed as a gavage. In addition to DMBA treatment as the second group, in the 3,4and 5 groups after cancer creation, respectively affected by alpha interferon (α-IFN),alpha interferon conjugated with single walled carbon nano tube (α-IFN-SWNT) and encapsulated in poly lactic poly glycolic acid (α-IFN-PLGA). Tumor marker was measured in recent three groups. Results: The ANOVA test was used to determine the differences among the groups. Cancer inducing in rats (group 2) caused a significant increase in blood levels of CA15-3 (P<0.05). Administration of α-IFN, α-IFN –SWNT and α-IFN-PLGA in 3 groups of cancerous rats caused a significant decrease in blood levels of CA15-3 only the group that treated with α-IFN-PLGA (p<0.05). Conclusion: the results of this study indicate that nano drugs more effective than traditional drug in cancer treatment, although further work is needed to elucidate the safety and side effect of these compound in human.

Keywords: breast cancer, nano drug, tumor markers, CA15-3, α-IFN-PLGA, -IFN –SWNT

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Authors: A. Satty, H. Mwambi

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Much clinical trials data-based research are characterized by the unavoidable problem of dropout as a result of missing or erroneous values. This paper aims to review some of the various techniques to address the dropout problems in longitudinal clinical trials. The fundamental concepts of the patterns and mechanisms of dropout are discussed. This study presents five general techniques for handling dropout: (1) Deletion methods; (2) Imputation-based methods; (3) Data augmentation methods; (4) Likelihood-based methods; and (5) MNAR-based methods. Under each technique, several methods that are commonly used to deal with dropout are presented, including a review of the existing literature in which we examine the effectiveness of these methods in the analysis of incomplete data. Two application examples are presented to study the potential strengths or weaknesses of some of the methods under certain dropout mechanisms as well as to assess the sensitivity of the modelling assumptions.

Keywords: incomplete longitudinal clinical trials, missing at random (MAR), imputation, weighting methods, sensitivity analysis

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Authors: Mustafa M. Donma, Orkide Donma

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Zinc is a vital element required for growth and development. This fact makes zinc important, particularly for children. It maintains normal cellular structure and functions. This essential element appears to have protective effects against coronary artery disease and cardiomyopathy. Higher serum zinc levels are associated with lower risk of cardiovascular diseases (CVDs). There is a significant association between low serum zinc levels and heart failure. Zinc may be a potential biomarker of cardiovascular health. High sensitive cardiac troponin T (hs-cTnT) and cardiac myosin binding protein C (cMyBP-C) are new generation markers used for prediagnosis, diagnosis, and prognosis of CVDs. The aim of this study is to determine zinc as well as new generation cardiac markers profiles in children with normal body mass index (N-BMI), obese (OB), morbid obese (MO) children, and children with metabolic syndrome (MetS) findings. The association among them will also be investigated. Four study groups were constituted. The study protocol was approved by the institutional Ethics Committee of Tekirdag Namik Kemal University. Parents of the participants filled informed consent forms to participate in the study. Group 1 is composed of 44 children with N-BMI. Group 2 and Group 3 comprised 43 OB and 45 MO children, respectively. Forty-five MO children with MetS findings were included in Group 4. World Health Organization age- and sex-adjusted BMI percentile tables were used to constitute groups. These values were 15-85, 95-99, and above 99 for N-BMI, OB, and MO, respectively. Criteria for MetS findings were determined. Routine biochemical analyses, including zinc, were performed. High sensitive-cTnT and cMyBP-C concentrations were measured by kits based on enzyme-linked immunosorbent assay principle. Appropriate statistical tests within the scope of SPSS were used for the evaluation of the study data. p<0.05 was accepted as statistically significant. Four groups were matched for age and gender. Decreased zinc concentrations were measured in Groups 2, 3, and 4 compared to Group 1. Groups did not differ from one another in terms of hs-cTnT. There were statistically significant differences between cMyBP-C levels of MetS group and N-BMI as well as OB groups. There was an increasing trend going from N-BMI group to MetS group. There were statistically significant negative correlations between zinc and hs-cTnT as well as cMyBP-C concentrations in MetS group. In conclusion, inverse correlations detected between zinc and new generation cardiac markers (hs-TnT and cMyBP-C) have pointed out that decreased levels of this physiologically essential trace element accompany increased levels of hs-cTnT as well as cMyBP-C in children with MetS. This finding emphasizes that both zinc and these new generation cardiac markers may be evaluated as biomarkers of cardiovascular health during severe childhood obesity precipitated with MetS findings and also suggested as the messengers of the future risk in the adulthood periods of children with MetS.

Keywords: cardiac myosin binding protein-C, cardiovascular diseases, children, high sensitive cardiac troponin T, obesity

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Authors: Y. I. Tretyakova, S. G. Shulkina, T. Y. Kravtsova, A. A. Antipova, N. Y. Kolomeets

Abstract:

The research aimed to assess the functional significance of tumor necrosis factor-alpha (TNF-α) gene polymorphism at the -308G/A (rs1800629) region and vascular endothelial growth factor A (VEGFA) gene polymorphism at the -634G/C (rs 2010963) region in the development of ulcerative colitis (UC), focusing on patients from the Perm region, Russia. We examined 70 UC patients and 50 healthy donors during the active phase of the disease. Our focus was on TNF-α and VEGF concentration in the blood serum, as well as TNF-α and VEGFA gene polymorphisms at the -308G/А and -634G/C regions, respectively. We found that TNF-α and VEGF levels were significantly higher in patients with severe UC and high endoscopic activity compared to those with milder forms of the disease and low endoscopic activity. These tests could serve as additional non-invasive markers for assessing mucosal damage in the large intestine of UC patients. The frequency of allele variations in the TNF-α gene -308G/A (rs1800629) revealed a significantly higher occurrence of the unfavorable homozygote AA in UC patients compared to donors. Additionally, the major allele G and the allele pair GG were more frequent in patients with mild to moderate disease and 1-2 degree of endoscopic activity than in those with severe UC and 3-4 degree of endoscopic activity (χ2=14.19; p=0.000). We also observed a mutant allele A and the unfavorable homozygote AA associated with severe progressive UC. The occurrence of the mutant allele increased the risk of severe UC by 5 times (OR 5.03; CI 12.07-12.21). We did not find any significant differences in the frequency of the CC homozygote (χ2=1.02; p=0.6; OR=1.32) and the mutant allele C of the VEGFA gene -634G/C (rs 2010963) (χ2=0.01; p=0.913; OR=0.97) between groups of UC patients and healthy individuals. However, we detected that the mutant allele C and the unfavorable homozygote CC of the VEGFA gene were associated with more severe endoscopic changes in the colonic mucosa of UC patients (χ2=25,76; р=0,000; OR=0,15). The presence of the mutant allele increased the risk of severe UC by 6 times (OR 6,78; CI 3,13–14,7). We found a direct correlation between TNF-α and VEGFA gene polymorphisms, increased production of the same factors, disease severity, and endoscopic activity (р=0.000). Therefore, the presence of the mutant allele A and homozygote AA of the TNF-α gene at the -308G/A region and the mutant allele C and homozygote CC of the VEGFA gene at the -634G/C region are associated with risks related to an unfavorable clinical course of UC, frequent recurrences, and rapid progression. These findings should be considered when making prognoses regarding the clinical course of the disease and selecting treatment strategies. The presence of the homozygote AA in the TNF-α gene (rs1800629) is considered a sign of genetic predisposition to UC.

Keywords: gene polymorphism, TNF-α, ulcerative colitis, VEGF

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Authors: Sujata Chaturvedi, Ishita Pant, Deepak Kumar Jha, Vinod Kumar Singh Gautam, Chandra Bhushan Tripathi

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Objective: To describe clinical, histopathological and immunohistochemical profile of glioblastoma in patients and to correlate these findings with patient survival. Material and methods: 30 cases of histopathologically diagnosed glioblastomas were included in this study. These cases were analysed in detail for certain clinical and histopathological parameters. Immunohistochemical staining for p53, epidermal growth factor receptor (EGFR), vascular endothelial growth factor (VEGF), mouse double minute 2 homolog (MDM2) and Ki67 was done and scores were calculated. Results of these findings were correlated with patient survival. Results: A retrospective analysis of the histopathology records and clinical case files was done in 30 cases of glioblastoma (WHO grade IV). The mean age of presentation was 50.6 years with a male predilection. The most common involved site was the frontal lobe. Amongst the clinical parameters, age of the patient and extent of surgical resection showed a significant correlation with the patient survival. Histopathological parameters showed no significant correlation with the patient survival, while amongst the immunohistochemical parameters expression of MDM2 showed a significant correlation with the patient survival. Conclusion: In this study incorporating clinical, histopathological and basic panel of immunohistochemistry, age of the patient, extent of the surgical resection and expression of MDM2 showed significant correlation with the patient survival.

Keywords: glioblastoma, p53, EGFR, VEGF, MDM2, Ki67

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Authors: Peter David Reiss

Abstract:

The concept of the psychosis prodrome has allowed for the identification of adolescent and young adult patients who have a significantly elevated risk of developing schizophrenia spectrum disorders. A number of different interventions have been tested in order to prevent or delay progression of symptoms. To date, there has been no consistent meta-analytical evidence to support efficacy of antipsychotic treatment for patients in the prodromal state, and their use remains therefore inconclusive. Although antipsychotics may manage symptoms transiently, they have not been found to prevent or delay onset of psychotic disorders. Furthermore, pharmacological intervention in high-risk individuals remains controversial, because of the antipsychotic side effect profile in a population in which only about 20 to 35 percent will eventually convert to psychosis over a two-year period, with even after two years conversion rates not exceeding 30 to 40 percent. This general estimate is additionally problematic, in that it ignores the fact that there is significant variation in individual risk among clinical high-risk cases. The current lack of reliable tests for at-risk patients makes it difficult to justify individual treatment decisions. Preventive treatment should ideally be dictated by an individual’s risk while minimizing potentially harmful medication exposure. This requires more accurate predictive assessments by using valid and accessible prognostic markers. The following will compare prediction and risk modification potential of behavioral biomarkers such as disturbances of basic sense of self and emotion awareness, neurocognitive biomarkers such as attention, working and declarative memory, and neurophysiological biomarkers such as glutamatergic abnormalities and NMDA receptor dysfunction. Identification of robust biomarkers could therefore not only provide more reliable means of psychosis prediction, but also help test and develop new clinical interventions targeted at the prodromal state.

Keywords: at-risk mental state, biomarkers, glutamatergic system, NMDA receptor, psychosis prodrome, schizophrenia

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Authors: Shirin jalili, Hadi Shirzad, Mahasti Modarresi, Samaneh Nabavi, Somayeh Khanjani

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Identifying the source tissue of biological material found at crime scenes can be very informative in a number of cases. Despite their usefulness, current visual, catalytic, enzymatic, and immunologic tests for presumptive and confirmatory tissue identification are applicable only to a subset of samples, might suffer limitations such as low specificity, lack of sensitivity, and are substantially impacted by environmental insults. In addition their results are operator-dependent. Recently the possibility of discriminating body fluids using mRNA expression differences in tissues has been described but lack of long term stability of that Molecule and the need to normalize samples for each individual are limiting factors. The use of DNA should solve these issues because of its long term stability and specificity to each body fluid. Cells in the human body have a unique epigenome, which includes differences in DNA methylation in the promoter of genes. DNA methylation, which occurs at the 5′-position of the cytosine in CpG dinucleotides, has great potential for forensic identification of body fluids, because tissue-specific patterns of DNA methylation have been demonstrated, and DNA is less prone to degradation than proteins or RNA. Previous studies have reported several body fluid-specific DNA methylation markers.The presence or absence of a methyl group on the 5’ carbon of the cytosine pyridine ring in CpG dinucleotide regions called ‘CpG islands’ dictates whether the gene is expressed or silenced in the particular body fluid. Were described methylation patterns at tissue specific differentially methylated regions (tDMRs) to be stable and specific, making them excellent markers for tissue identification. The results demonstrate that methylation-based tissue identification is more than a proof-of-concept. The methodology holds promise as another viable forensic DNA analysis tool for characterization of biological materials.

Keywords: DNA methylation, forensic science, epigenome, tDMRs

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Authors: Rafia S. Al-lamki, Jun Wang, Simon Pacey, Jordan Pober, John R. Bradley

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Tumor necrosis factor alpha (TNF), signaling through TNFR2, may act an autocrine growth factor for renal tubular epithelial cells. Clear cell renal carcinomas (ccRCC) contain cancer stem cells (CSCs) that give rise to progeny which form the bulk of the tumor. CSCs are rarely in cell cycle and, as non-proliferating cells, resist most chemotherapeutic agents. Thus, recurrence after chemotherapy may result from the survival of CSCs. Therapeutic targeting of both CSCs and the more differentiated bulk tumor populations may provide a more effective strategy for treatment of RCC. In this study, we hypothesized that TNFR2 signaling will induce CSCs in ccRCC to enter cell cycle so that treatment with ligands that engage TNFR2 will render CSCs susceptible to chemotherapy. To test this hypothesis, we have utilized wild-type TNF (wtTNF) or specific muteins selective for TNFR1 (R1TNF) or TNFR2 (R2TNF) to treat either short-term organ cultures of ccRCC and adjacent normal kidney (NK) tissue or cultures of CD133+ cells isolated from ccRCC and adjacent NK, hereafter referred to as stem cell-like cells (SCLCs). The effect of cyclophosphamide (CP), currently an effective anticancer agent, was tested on CD133+SCLCs from ccRCC and NK before and after R2TNF treatment. Responses to TNF were assessed by flow cytometry (FACS), immunofluorescence, and quantitative real-time PCR, TUNEL, and cell viability assays. Cytotoxic effect of CP was analyzed by Annexin V and propidium iodide staining with FACS. In addition, we assessed the effect of TNF on isolated SCLCs differentiation using a three-dimensional (3D) culture system. Clinical samples of ccRCC contain a greater number SCLCs compared to NK and the number of SCSC increases with higher tumor grade. Isolated SCLCs show expression of stemness markers (oct4, Nanog, Sox2, Lin28) but not differentiation markers (cytokeratin, CD31, CD45, and EpCAM). In ccRCC organ cultures, wtTNF and R2TNF increase CD133 and TNFR2 expression and promote cell cycle entry whereas wtTNF and R1TNF increase TNFR1 expression and promote cell death of SCLCs. Similar findings are observed in SCLCs isolated from NK but the effect was greater in SCLCs isolated from ccRCC. Application of CP distinctly triggered apoptotic and necrotic cell death in SLCSs pre-treatment with R2TNF as compared to CP treatment alone, with SCLCs from ccRCC more sensitive to CP compared to SLCS from NK. Furthermore, TNF promotes differentiation of SCLCs to an epithelial phenotype in 3D cultures, confirmed by cytokeratin expression and loss of stemness markers Nanog and Sox2. The differentiated cells show positive expression of TNF and TNFR2. These findings provide evidence that selective engagement of TNFR2 drive CSCs to cell proliferation/differentiation, and targeting of cycling cells with TNFR2 agonist in combination with anti-cancer agents may be a potential therapy for RCC.

Keywords: cancer stem cells, ccRCC, cell cycle, cell death, TNF, TNFR1, TNFR2, CD133

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Authors: Iman Nezamzadeh, Ali Faghih, Behnam Oveisi

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In order to reduce earthquake hazards in urban areas, it is necessary to perform comprehensive studies to understand the dynamics of the active faults and identify potentially high risk areas. The fault slip-rates in Late Quaternary sediment are critical indicators of seismic hazard and also provide valuable data to recognize young crustal deformations. To measure slip-rates accurately, is needed to displacement of geomorphic markers and ages of quaternary sediment samples of alluvial deposit that deformed by movements on fault. In this study we produced information about Intermediate term slip rate of Sabzpushan Fault Zone (SPF) within the central part of the Zagros Mountains of Iran using OSL dating technique to make better analysis of seismic hazard and seismic risk reduction for Shiraz city. For this purpose identifiable geomorphic fluvial surfaces help us to provide a reference frame to determine differential or absolute horizontal and vertical deformation. Optically stimulated luminescence (OSL) is an alternative and independent method of determining the burial age of mineral grains in Quaternary sediments. Field observation and satellite imagery show geomorphic markers that deformed horizontally along the Sabzpoushan Fault. Here, drag folds is forming because of evaporites material of Miocen Formation. We estimate 2.8±0.5 mm/yr (mm/y) horizontal slip rate along the Sabzpushan fault zone, where ongoing deformation is involve with drug folding. The Soltan synclinal structure, close to the Sabzpushan fault, shows slight uplift rate due to active core-extrousion.

Keywords: slip rate, active tectonics, OSL, geomorphic markers, Sabzpushan Fault Zone, Zagros, Iran

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Authors: Wafaa M. El- Kotb, Soheir Mohamed Weheida, Manal E. Fareed

Abstract:

The aim of this study was to determine the effect of implementing a teaching module about diet and exercises on clinical outcomes of patients with gout. Subjects: A purposive sample of 60 adult gouty patients was selected and randomly and alternatively divided into two equal groups 30 patients in each. Setting: The study was conducted in orthopedic out patient's clinic of Menoufia University. Tools of the study: Three tools were utilized for data collection: Knowledge assessment structured interview questionnaire, Clinical manifestation assessment tools and Nutritional assessment sheet. Results: All patients of both groups (100 %) had poor total knowledge score pre teaching, while 90 % of the study group had good total knowledge score post teaching by three months compared to 3.3 % of the control group. Moreover the recovery outcomes were significantly improved among study group compared to control group post teaching. Conclusion: Teaching study group about diet and exercises significantly improved their clinical outcomes. Recommendation: Patient's education about diet and exercises should be ongoing process for patients with gout.

Keywords: clinical outcomes, diet, exercises, teaching module

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Authors: Asma Naseer Cheema, Attya Bhatti, Jabar Ali, John Peter

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Background: High cholesterol levels, endothelial dysfunction, inefficient coagulation cascade and hyper inflammatory response all are the basis of coronary artery disease (CAD). Several studies are carried out to see the genetic influence of these factors on disease outcome. Objective: The objective of our study was to see the association of 10 putative loci with coronary artery disease in our population. Materials & Methods: We screened our population for 10 putative loci of CAD showing significant association (p < 5x10-8) with candidate genes (regulating the cholesterol metabolism, endothelial function, coagulation cascade and inflammatory response of body). Hardy-Weinberg equilibrium and linkage disequilibrium in cases and controls s were estimated separately. Approximately 5-10 ng of dried DNA in 384 well plate format was used to genotype each sample on the Sequenom iPLEX assay at University of Pittsburgh Genomics and Proteomics Core Laboratories. It was built on single-base primer extension with the MALDI-TOF MS detection possessing high sensitivity and specificity. The SNPs were genotyped through Taqman assay. Hardy Weinberg test was applied. The 10 SNPs were selected as genetic markers for this study (rs579459, rs1561198, rs2954029, rs1122608, rs17114036, rs9515203, rs10947789, rs7173743, rs2895811, rs2075650). Results: Mean age of the patient was 52 ± 11 years. Blood pressure and positive family history was found a significant risk factor for CAD. None of the selected SNPs showed significant association with coronary artery disease in our population (p>0.05). Conclusion: rs579459, rs1561198, rs2954029, rs1122608, rs17114036, rs9515203, rs10947789, rs7173743, rs2895811, rs2075650 are not significant genetic markers for CAD in our population.

Keywords: CAD, genetic markers, loci, risk factors

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