ISSN  2587-2362  |  E-ISSN  2618-642X
International Journal of Medical Biochemistry - Int J Med Biochem : 7 (2)
Volume: 7  Issue: 2 - 2024
1.Front Matter

Pages I - X

2.25-hydroxyvitamin-D3 analysis with high-performance liquid chromatography in terms of total analytical error and measurement uncertainty
Muhammed Seyithanoğlu
doi: 10.14744/ijmb.2024.81894  Pages 45 - 50
INTRODUCTION: Ensuring the accuracy and consistency of data obtained in the biochemistry laboratory is essential for obtaining reliable and comparable results. This study aims to calculate total analytical error (TAE) and measurement uncertainty (MU) values to assess the analytical performance of the 25-hydroxyvitamin D3 (25-OH vitD3) analyte measured using high-performance liquid chromatography (HPLC) in our laboratory
METHODS: In our study, the internal quality control (IQC) results, which were analyzed at two levels daily between 01–01.2022–31.12.2022, and the data of the external quality control (EQC) program, which was performed at two levels per period for four periods per year, were retrospectively examined for the MU and TAE calculations of the 25-OH vitD3 analyte. TAE was calculated by the formula TAE% = Bias% + (1.65×CV%). MU has been calculated adhering to the Nordtest guideline.
RESULTS: In our study, while TAE values and U value calculated using EQC data for bias and u(bias) calculation were found to be higher than the analytical performance goals we used in our study, TAE values and U value calculated using IQC data for bias and RMSbias calculation were found to be lower than the analytical performance goals we used in our study.
DISCUSSION AND CONCLUSION: Clinical laboratories should evaluate analytical performance at regular intervals using appropriate methods. In cases where the number of participants using the same method and device in the EQC program is low, we recommend that EQC data not be used in the calculation of the bias component when evaluating analytical performance with TAE or MU.

3.Bioinformatic assessment of the relationship between breast cancer and autophagy-related protein Ambra1 mutation
Umut Karabay, Durmuş Ayan
doi: 10.14744/ijmb.2024.59244  Pages 51 - 59
INTRODUCTION: Autophagy protein 1, regulated by Beclin 1 (Ambra1), promotes tumor formation and development by modulating autophagy. Therefore, in situ intervention in autophagy is a promising new strategy for tumor therapy. We aimed to evaluate the possible effects of changes in the Ambra1 gene on breast cancer (BC) treatment in the BRCA cohort.
METHODS: The gene profile of a total of 996 patients with BC was examined using data obtained from the Cancer Genome Atlas database via cBioPortal. The effects of mutations on proteins were examined by scoring the Polymorphism Phenotyping v2, Mutation Assessor, and Sorting Intolerant from Tolerant databases. The association of genes with other genes was determined with the STRING database. Kaplan-Meier Plot database was used by evaluating the overall survival (OS). The promoter methylation was evaluated by the UALCAN database.
RESULTS: Eleven mutations were detected. Four of these mutations were truncated proteins. Ambra1 tissue expression levels were upregulated compared to healthy tissue in the BRCA cohort; this was not statistically significant (p>0.05).
Decreased Ambra1 expression levels were associated with a shorter OS (p=0.038). Ambra1 promoter region hypermethylation was significant in the BRCA cohort compared to healthy tissue (p<0.001).
DISCUSSION AND CONCLUSION: To our best knowledge, our study is the first to examine the relationship between BC and Ambra1 using bioinformatic tools. Ambra1 may be a candidate target molecule within the treatment strategy due to the mutations evaluated in the BRCA cohort, hypermethylation status, and the association of Ambra1 with shorter OS. However, these situations need to be confirmed by further studies.

4.Relationship between atherogenic index of plasma, asprosin, and metrnl levels in hemodialysis patients
Meltem Yardım, Levent Deniz, Mehmet Akif Saltabas, Damla Kayalp, Ramazan Fazıl Akkoç, Tuncay Kuloğlu
doi: 10.14744/ijmb.2024.55376  Pages 60 - 66
INTRODUCTION: This study aimed to evaluate the relationship between the atherogenic index of plasma (AIP) and plasma asprosin and metrnl levels in hemodialysis (HD) patients.
METHODS: Forty-eight patients receiving HD treatment with a diagnosis of end-stage renal disease (ESRD) were included. The control group comprised 34 age-, sex-, and body mass index-matched healthy volunteers without a history
of renal disease. ESRD patients were divided into two groups: high-risk (AIP≥0.24) and low-moderate risk (AIP<0.24).
Asprosin and metrnl levels in the plasma of blood samples taken just before dialysis were studied by enzyme-linked immunosorbent assay.
RESULTS: A significant difference was found between the control group [23.3(19.9-27.7 ng/mL)], the low-moderate risk
group [39.3(34.9-40.8 ng/mL)], and the high-risk group [48.1(44.5-49.9 ng/mL)] in terms of asprosin levels (for each p<0.001). Asprosin values of both low-moderate risk and high-risk groups were significantly higher than the controls.
In the high-risk group, plasma asprosin levels were higher than in the low-moderate risk group (p=0.012). Metrnl levels of the high-risk group were found to be lower than both the control and low-risk groups (p<0.001 and p=0.003, respectively). AIP showed a positive relation to asprosin and a negative relation to metrnl.
DISCUSSION AND CONCLUSION: Logistic regression analysis has revealed important insights into the independent relationships between metrnl, asprosin, and high AIP values in HD patients. These findings support the anti-atherogenic potential of metrnl and suggest the potential atherogenic effects of asprosin, highlighting the complex interplay between adipokines and cardiovascular risk in this patient population.

5.Evaluation of drug abuse test analysis: One year experience
Gamze Gök, Turan Turhan
doi: 10.14744/ijmb.2024.46330  Pages 67 - 72
INTRODUCTION: Drug abuse is a major problem for public health and it has negative impacts on people's health. Drug analysis methods used in our country vary, but the results show that drug abuse is an increasing problem in our country compared to previous years. In our hospital, drug urine screen tests are performed to analyze amphetamines, benzodiazepines, cannabinoids, cocaine, and their metabolites, opiates, and synthetic cannabinoids. Our research aimed to evaluate the frequency of drug use according to age and gender.
METHODS: A total of 2172 amphetamine, 2172 benzodiazepine, 2172 cannabinoids, 2169 cocaine and cocaine metabolites, 2168 opiates, and 1906 synthetic cannabinoid analysis results were included in our study. Analyses were performed by Advia® (Siemens Healthineers, Erlangen, Germany) autoanalyzer with a homogeneous immunoassay method that enables qualitative or semi-quantitative analysis of analytes. At every stage of our study, we worked in accordance with the Declaration of Helsinki.
RESULTS: The results were as follows: 1989 (91.6%) were negative, 183 (8.4%) were positive for amphetamine, 1924 (88.6%) were negative, and 248 (11.4%) were positive for benzodiazepine; 2057 (94.7%) were negative and 115 (5.3%) were positive for cannabinoids; 2157 (99.4%) were negative and 12 (0.6%) were positive for cocaine and its metabolites; and 1865 (97.8%) were negative and 41 (2.2%) were positive for synthetic cannabinoids. Males' positive results for amphetamine (p<0.001), cannabinoid (p<0.001), and opiates (p=0.026) were statistically significant when compared according to gender. The adult group's positive results for amphetamine (p<0.001), cannabinoid (p<0.001), and synthetic cannabinoids (p=0.046) were statistically significant when compared according to age.
DISCUSSION AND CONCLUSION: Sharing drug positivity situations in different age groups and genders may help to draw attention to this problem and maybe preventive. More studies including more than one year of results may be beneficial.

6.IL28B rs12979860 gene polymorphism and sofosbuvir-based therapy response in HCV-infected Pakistani patients
Maleha Asim, Anoshia Javed, Ammarah Saeed, Bushra Adeel, Shazia Siddique, Saddaf Ayub
doi: 10.14744/ijmb.2024.75437  Pages 73 - 80
INTRODUCTION: Interleukin-28B (IL-28B) gene polymorphisms play an important role in response prediction of direct-acting antivirals (DAAs) treatment, including Sofosbuvir and Daclatasvir with or without Ribavirin. The purpose of this study was to assess the IL-28B polymorphism SNP (rs12979860) and other clinical factors as response predictors for the sustained virological response (SVR) in chronic HCV-infected patients taking DAA therapy.
METHODS: A cross-sectional and observational study was carried out among 104 HCV-infected patients who completed a course of Sofosbuvir and Daclatasvir along with Ribavirin. Patients were classified according to their response to therapy. Genotyping of IL-28B was determined through polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay, and HCV genotyping was identified by PCR method. We analyzed the response prediction of IL-28 gene polymorphism among patients receiving DAA therapy.
RESULTS: Overall, IL-28B CC, CT, and TT genotypes were found in 56 (53.8%), 22 (21.2%), and 26 (25.0%) patients, respectively. Higher early virological response (EVR) and SVR were observed in patients with the rs12979860 CC alleles (82.1% and 75%) as compared to CT/TT alleles (54.2% and 20.8%). IL-28B CC genotype (OR=0.14; 95% CI=0.04-0.44; p=0.001) and EVR (OR=0.20; 95% CI=0.05-0.71; p=0.013) remained significantly associated with SVR in the multivariate regression analysis. However, the FIB-4 score (OR=4.24; 95% CI=1.46-11.75; p=0.008) is a strong predictor of non-SVR.
DISCUSSION AND CONCLUSION: The antiviral efficacy of triple therapy (sofosbuvir, daclatasvir, and ribavirin) is influenced by the variability of the IL-28B gene, as well as the EVR and FIB-4 score. These variables also play a significant role in predicting the treatment response of patients with chronic HCV infection in Pakistan.

7.Investigation of Prothrombin Time, International Normalized Ratio and Activated Partial Thromboplastin Time reference ranges in children
Gözde Ülfer
doi: 10.14744/ijmb.2024.77527  Pages 81 - 86
INTRODUCTION: This study aimed to ascertain pediatric age-specific reference ranges for prothrombin time (PT), international normalized ratio (INR), and activated partial thromboplastin time (aPTT). Retrospective data were obtained from healthy children who had undergone preoperative tests and compared with those obtained from a group of adult patients.
METHODS: Reference individuals were determined by the indirect method. A total of 15,179 patients who presented to our hospital in 2022 and 2023 were retrospectively reviewed. Pediatric patients were divided into three age groups: 1–5 years (n=1949), 6–10 years (n=1563), and 11–17 years (n=508). The adult age group consisted of healthy individuals aged 18–50 years (n=11165). The tests were run in a coagulation autoanalyzer with the mechanical coagulometric measurement method. Reference ranges were analyzed using the non-parametric method statistically.
RESULTS: The mean PT, INR, and aPTT values were found to be 14.34±1.99 s, 1.07±0.16, and 31.43±3.47 s, respectively, in children aged 1–5 years; 14.48±1.40 s, 1.08±0.11, and 31.30±2.66 s, respectively, in those aged 6–10 years; and 14.73±1.12 s, 1.10±0.09, and 31.21±2.91 s, respectively, in those aged 11–17 years. Among the adults aged 18–50 years, the mean PT, INR, and aPTT values were 13.95±1.40 s, 1.04±0.11, and 30.21±2.86 s, respectively. The mean PT, INR, and aPTT values of children aged 1–5 years, 6–10 years, and 11–17 years were statistically significantly higher than those of adults aged 18–50 years (for each group p<0.01).
DISCUSSION AND CONCLUSION: It is important for laboratories to employ age-specific reference ranges for coagulation tests performed on children to ensure accurate diagnosis and avoid unnecessary further investigations. In this study, the reference ranges of the PT, INR, and aPTT parameters were determined for pediatric patients and found to be significantly higher than those of adults, which will be useful for clinical evaluation and diagnosis.

8.Assessment of IL-6, TGF-β1 and CTX-II in the diagnosis of early Post-Traumatic Osteoarthritis of knee
Keerthy Rethinam Meenakshisundaram, Thiagarajan Keddin Alwar, Ramchand Nannapan, Vellora Mohanakrishnan Vinodhini, Karthikeyan Rajamani, Kuppusamy Mahesh Kumar, Santhi Silambanan
doi: 10.14744/ijmb.2024.86158  Pages 87 - 94
INTRODUCTION: Osteoarthritis is an inflammatory and degenerative disorder characterized by the degradation of the extracellular matrix. It commences with injuries to the knee that activate inflammatory pathways. Trauma may predispose to osteoarthritis, called post-traumatic osteoarthritis (PTOA). The disease is diagnosed in the later stages with (KL grade>2) as seen by X-ray; measuring the biomarkers present in the body fluids holds significant promise for the early detection and evaluation of PTOA. The study aimed to establish the role of inflammatory and collagen markers such as serum interleukin 6 (IL-6), transforming growth factor beta 1 (TGF-β1), and urine C-terminal cross-linked telopeptide of type II collagen (CTX-II) in the diagnosis of early post-traumatic osteoarthritis of the knee.
METHODS: This case-control study was conducted among 80 participants, of which 40 were apparently healthy individuals, and 40 were cases with a history of trauma to the knee joint in the past ten to 12 weeks. Baseline characteristics, body mass index (BMI), Visual Analog Score (VAS), and Western Ontario McMasters Universities Osteoarthritis Index (WOMAC) were collected from all the participants. X-ray and MRI were done in the cases. Serum IL-6 and TGF-β1, and urine CTX-II were analyzed by ELISA. Statistical analysis was done with SPSS version 16. A P value ≤ 0.05 was considered statistically significant.
RESULTS: The mean serum IL-6, TGF-β1, and urine CTX-II levels were significantly higher in cases than in controls, with P values of 0.025, 0.033, and 0.040 respectively. IL-6 showed correlations with age, WOMAC score, and urine CTX-II values. TGF-β1 showed a positive correlation with VAS.
DISCUSSION AND CONCLUSION: Individuals with previous knee joint trauma exhibited notably elevated serum IL-6, TGF-β1, and urine CTX-II levels. Among the three biomarkers, IL-6 seemed to be a potential biomarker of early post-traumatic osteoarthritis in patients with knee injuries.

9.Creatinine normalization approach to diluted urine samples screened by LC-MS/MS method
Cigdem Karakukcu, Derya Kocer, Veysel Uzen, Hatice Saracoglu
doi: 10.14744/ijmb.2024.57441  Pages 95 - 100
INTRODUCTION: Urine is the most used matrix in drug analysis; however, it is susceptible to adulteration or tampering. Urine creatinine is the most important urine integrity parameter used as an indicator of dilution. This study aimed to evaluate the prevalence of diluted urine samples and the change in positivity after creatinine normalization.
METHODS: Urine samples screened by the LC-MS/MS method over a 3.5-year period (n=21,927) were included in the study. Positivity rates were evaluated in both total and diluted urine samples. Additionally, the impact of creatinine normalization on samples with substance concentrations above the limit of quantitation (LOQ) and below the cut-off was investigated.
RESULTS: A total of 350,832 tests were conducted on 21,927 urine samples, resulting in an overall positivity rate of 21.2% (n=4652). The ratio of diluted urine was 1.6% (n=343), with 61.5% (n=211) testing negative (<LOQ), 23.3% (n=80) testing positive (at least one substance >cut-off), and 15.2% (n=52) testing above LOQ and below cut-off. After creatinine normalization in diluted urines, the sample positivity rate increased from 23.3% (n=80) to 33.8% (n=116) (p<0.001), and the substance positivity rate increased from 2.3% (n=125) to 3.9% (n=212) (p<0.001).
DISCUSSION AND CONCLUSION: Precautions should be taken in reporting diluted urine samples to avoid reporting false negative results. The creatinine normalization approach shows promise in laboratories using quantitative screening methods such as LC-MS/MS for samples with substance concentrations above the LOQ and below the cut-off. However, more clinical and laboratory collaboration is needed for its routine application.

10.Unveiling the latest insights into Androgen Receptors in Prostate Cancer
Suleiman Zakari, Christogonus Chichebe Ekenwaneze, Emmanuel Chimuebuka Amadi, Abrar Abuhamdia, Olubanke Olujoke Ogunlana, Suleiman Zakari
doi: 10.14744/ijmb.2024.93585  Pages 101 - 113
Prostate cancer (PCa) is a major cause of cancer-related mortality worldwide, with a rising incidence observed over the years. The androgen receptor (AR) signaling pathway plays a pivotal role in male development and maintaining masculine characteristics. Dysregulation of AR signaling in PCa can lead to disease progression and resistance to standard therapies. Understanding the intricate regulation and function of AR in both healthy and diseased states is crucial for developing effective treatment strategies. This review comprehensively explores the role of androgen receptors in PCa susceptibility, disease progression, and treatment response by analyzing recent literature. An extensive search of peer-reviewed publications in major databases, including PubMed, Scopus, and Web of Science, was conducted using specific keywords related to androgen receptor, prostate cancer, disease progression, and treatment resistance. Relevant conference abstracts and clinical trial reports were also included. The review presents an overview of the role of androgen receptors in PCa initiation, progression, and treatment resistance. It also highlights the role of SPOP as an emerging biomarker associated with AR signaling dysregulation and their potential utility for early detection and personalized treatment approaches. Additionally, recent advances in targeting the AR pathway for novel therapeutic strategies to improve patient outcomes and overcome treatment resistance in advanced PCa are discussed. The findings contribute to a comprehensive understanding of the AR signaling pathway in PCa and offer insights into its multifaceted role in disease development and treatment response. They may pave the way for innovative therapeutic interventions and precision medicine approaches based on specific AR signaling profiles, enhancing patient care and reducing the burden of this lethal disease.

11.Fibroblast growth factors: properties, biosynthesis, biological functions, therapeutic applications and engineering
Nitin Tiwari, Ashok Tiwari, Lalita Mehra, Arjun Ganguly, Kamlesh Darji, Muzafar Pandit, Rachana R
doi: 10.14744/ijmb.2024.16768  Pages 114 - 126
Fibroblast Growth Factors (FGFs) function as signaling molecules within various signaling pathways, regulating the proliferation, migration, and differentiation of soft connective tissues, nerves, epithelial tissue, and bone. The FGF family comprises 22 members, with acidic Fibroblast Growth Factor (aFGF/FGF-1) and basic Fibroblast Growth Factor (bFGF/FGF-2) being of primary significance. This article explores the biochemical and biological properties of different FGFs, elucidating their roles in various biological processes. Additionally, it delves into the interactions between FGFs and Receptor tyrosine kinases (RTKs), which activate several cell signaling cascades, such as the RAS/MAPK (Mitogen-activated Protein Kinase) pathway, PI3K (phosphoinositide 3-kinase)/AKT (v-akt murine thymoma viral oncogene homolog) pathway, PLC-γ (Phospholipase C-γ) pathway, and Signal Transducer and Activator of Transcription (STAT) pathway, to facilitate diverse cellular functions. The article also examines methods for engineering FGFs, including N-terminal truncation, point mutations, or combinations thereof, for therapeutic applications in tissue regeneration, angiogenesis, and repairing damaged tissues such as cartilage, bone, ligaments, and skin. Finally, it concludes with a discussion of the delivery systems for FGFs, encompassing scaffolds, hydrogels, as well as nano- and micro-particulate methods.

12.The changes of oxidative stress markers and Vitamin E in patients with diabetes using SGLT2 inhibitors
Pradnya Padalkar
doi: 10.14744/ijmb.2024.68926  Pages 127 - 128
Abstract | Full Text PDF

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