Journal of Advanced Biotechnology and Experimental Therapeutics

326-336

Authors: Mirna Kifah Faiq , Eman Saadi Saleh , Omar Basher Fathalla

The high mobility group A1 gene (HMGA1) rs139876191 variant has been related to metabolic syndrome and type 2 diabetes, but data are lacking in Middle Eastern populations. The study aimed to assess whether the HMGA1 rs139876191 variant is associated with metabolic syndrome risk and whether this variant predicts the risk of insulin resistance. This case-control study was carried out at single center in Kirkuk city/ Iraq from February to August 2022. Polymorphisms in HMGA1 and genotyping were identified by Sanger sequencing of genomic DNA obtained from 91 Iraqi participants (61 patients with metabolic syndrome and 30 control). Lipid profile, serum (glucose and insulin), glycated hemoglobin, blood pressure, body mass index, and waist circumference were also measured. The high prevalence of the del/del genotype of rs139876191 was found. Minor allele frequency of rs139876191 was 0.16 in both metabolic syndrome and the control group. A non-significant difference in genotyping was identified between total metabolic syndrome and the control group. The del/ins variant was associated with significantly higher waist circumference, triglycerides (TG), low-density lipoprotein (LDL), very low-density lipoprotein (VLDL), and glycated hemoglobin (HbA1c) (P=0.03, 0.041, 0.007, 0.034, and 0.001, respectively), and significantly lower high-density lipoprotein (HDL) (p=0.000). Linear regression analysis showed no significant effect of the variant (del/ins) on developing insulin resistance. Thus, rs139876191 polymorphism with del/ins genotype in the HMGA1 gene was not associated with metabolic syndrome risk but it was associated with indices of metabolic syndrome including waist circumference, TG, HDL, LDL, VLDL, and HbA1c. Besides, this variant did not predict the risk of insulin resistance.

350-358

Authors: Mulia I.C. Pertiwi , Manik R. Wahyunitisari , Rebekah J. Setiabudi , Agung D.W. Widodo , Nurul Wiqoyah , M. Vitanata Arifijanto

Uropathogenic Escherichia coli (UPEC) is a fundamental cause of nosocomial urinary tract infection (UTI). UPEC can form biofilms on the mucosa of the urinary bladder and the surface of urethral catheters, thus causing clinical problems. Therefore, the current study aimed at the potential of synbiotics to manage biofilm-associated UTIs by analyzing biofilm inhibition UPEC with cell-free supernatant (CFS) of synbiotics. Biofilm inhibition was accomplished by inoculating each microbial suspension into 96-well microplates on tryptic soy broth medium at 37°C for 48 h and a microtiter plate reader was used at 595 nm to read the OD value. The outcome (%) was calculated from the OD value of CFS-treated with UPEC. The result of this research was that each CFS of synbiotic treatment displayed significantly different (P<0.05) results and was able to inhibit UPEC biofilm. The highest percentage of biofilm inhibition of UPEC was shown in CFS 8 treatment with a value of 41.51 ± 0.687, where CFS of synbiotic from Lactobacillus rhamnosus bacteria with 2% inulin. The lowest percentage of biofilm inhibition UPEC was shown in CFS 1 treatment with a value of 36.56 ± 1.987, where CFS from Lactobacillus plantarum bacteria with 0% inulin. It could be concluded that the higher the concentration of inulin in the CFS of synbiotics, the higher the percentage value of biofilm inhibition on UPEC, which indicates the potentials to manage or prevent UPEC-induced biofilm infection.

375-385

Authors: Mohanad Hasan , Reza Safaralizadeh , Mohammad Khalaj Kondori , Saeid Latifi Navid

Gastric cancer (GC), which is the fourth most prevalent cancer in the world is significantly threatened the health of people, particularly those in developing nations. Nearly all significant pathological and physiological mechanisms, including apoptosis, proliferation, cell cycle, differentiation, as well as DNA damage, are regulated by miRNAs. This study investigated the miR-221-3p expression and identified its target genes in GC tissue samples and cell lines, for an understanding of the miR-221-3p influence and basic processes in the progression of GC. GC tissues and matched marginal tissues were taken from 50 patients undergo gastric surgery. MiR-221-3p mimics, inhibitors, and negative controls (NC) were transfected into MKN-45 cells, using Lipofectamine RNAiMAX reagent. The proliferation was assessed by the MTT assay. Cell migration and invasion was assessed by Transwell assay. By combining Western blotting and qRT-PCR, the impact of miR-221-3p in the PIK3R1 expression in gastric cancer cells was examined. Overexpression of miR-221-3p significantly enhanced the migration, invasion, and proliferation of gastric cancer cells, conversely, transfection of miR-221-3p inhibitor led to opposite effect caused by overexpression of this miRNA on phenotypic characteristics of gastric cancer cell line. Additional investigation revealed that PIK3R1 was downregulated significantly by overexpression of miR-221-3p. Whereas, when the MKN-45 cells transfected with miR-221-3p inhibitor, PIK3R1 was noticeably overexpressed. Our current data indicate that miR-221-3p possibly work as a tumor promoter in the development of gastric cancer by negatively regulating PIK3R1 expression, hence miR-221-3p/ PIK3R1 highlighted as promising therapeutic targets or prognostic and diagnostic biomarkers for GC patients.

403-411

Authors: Raid D. Hashim , Israa Nathir , Esraa Ghazy , Ihsan Al Timimi , Tiba M. Hameed , Mayssam Hussein Mohammed Ali

Vitamin D deficiency is a highly prevalent medical condition associated with various clinical and biochemical outcomes although these outcomes might be absent in a significant percentage of patients. Altered serum calcium, phosphorus (PO4), parathyroid hormone (PTH) and alkaline phosphatase (ALP) activity are expected but not proven by various research. The current study aims to investigate the possibility of these biomarkers to be used as indicators of vitamin D deficiency or insufficiency. The study enrolled 150 randomly selected participants who have no acute or chronic medical condition with various levels of serum vitamin D ranging from normal to severely deficient. They were investigated for serum total calcium, PO4, PTH, and ALP activity. The results were statistically compared among the studied groups and correlated with serum vitamin D levels. There was no significant difference in mean serum calcium, PO4, and ALP activity among the groups with poor correlation with vitamin D. Mean serum PTH has shown a significant difference among the studied groups with a strong negative correlation with vitamin D. This difference was apparent even in patients with vitamin D insufficiency. Serum calcium, PO4 and ALP activity seem to have a poor correlation with vitamin D concentration. In addition, serum PTH might be considered a sensitive marker of both vitamin D deficiency and insufficiency.

419-428

Authors: Nishat Akther , Farjana A. Hira , Ayesha Khatun , Md. Shawan , Shaike M. Abdullah , Esrat J. Shorna , Ashekul Islam , Khairul Islam , Nazmul M. Hossain

Antimicrobial resistance has become an alarming issue all over the world. Moreover, in the developing countries as well as in Bangladesh facing this badly due to lack of study. Antibiotic resistance is prevalent among the bacterial pathogens causing urinary tract infections. Urinary tract infections affect >30% of people around the world. Worldwide, about 150 million people experience urinary tract infections each year. Antimicrobial treatment is often prescribed ‘empirically’ without antibiotic susceptibility testing to treat urinary tract infections (UTIs). The prevalence of antibiotic resistance is worsening, probably because of the increasing number of multidrug-resistant bacterial strains and the indiscriminate use of broad-spectrum antibiotics and empirical treatment. This study aims to determine the frequency and antibiotic resistance pattern of bacteria from suspected UTIs in Bangladeshi patients. In our study, about 200 samples of positive UTIs patients were analyzed with several parameters to see the antibiotic resistance rate. After examining the patient’s sample of UTIs antibiotic resistance rate is alarming and the most causative bacteria is Escherichia coli which is about 61.8%. Among the 200-urine sample suspected UTI patients 66% and 30% are female and male, respectively. Empirical therapy should be guided to control the antibiotic resistance. Especially, developing country should accept the challenge with proper infrastructure to get the surveillance of resistance, as resistance varies from region to region. Appropriate antibiotics should be focused for the rational use of antibiotics to prevent antibiotics resistance through common infections.

436-444

Authors: Riski Sulistiarini , Anggie Puranti , Wisnu Cahyo Prabowo

Hemorrhoids occur due to inflammation of the rectoanal venous plexus, which causes inflammation, pain, and can cause lumps. This inflammation may cause difficulty in defecation and may even cause heavy bleeding. Empirically, Tapak Liman (Elephantopus scaber L.) leaves have been used to treat hemorrhoids and have been known to contain flavonoids and terpenoid lactones which are active as anti-inflammatory agents. To determine the anti-hemorrhoidal activity of E. scaber leaves in reducing inflammation in hemorrhoids in Wistar rats, the research begins with the extraction of E. scaber leaves with 70% ethanol as solvent. Extracts were analyzed for total flavonoid content with quercetin standard. Anti-hemorrhoidal activity was measured based on the degree of edema formed based on the weight of the rectum through the surgical process. Hemorrhoid was induced with croton oil for 3 days on the rectoanal area in rats. The extracts were treated at concentration of 50, 100, and 200 mg/kgW for 7 days. The drug control group used oral diclofenac sodium for 7 days. E. scaber leaf ethanol extract contains alkaloids, flavonoids, saponins, and polyphenols. The flavonoid level was 3.433 mgQE/g extract. This study showed that E. scaber leaf extract had the best anti-hemorrhoidal activity at a dose of 100 mg/kgW of extract with a significance value of p<0.05. Also, the extract of E. scaber leaves containing flavonoids provides a fairly good anti-hemorrhoid activity compared to the control drug diclofenac sodium. It is suggested that this plant extract might be used as a new herbal anti-hemorrhoid drug.

458-467

Authors: Siti Fatkhul Jannah , Agung Dwi Wahyu Widodo , Rebekah Juniati Setiabudi

An intravenous catheter is a medical device used to inject intravenous fluid into the body. This procedure can cause coagulase-negative staphylococci (CoNS) bacteria such as Staphylococcus epidermidis penetrates the skin and forms biofilm on the catheter. Biofilms bring serious problems such as antibiotic resistance, the long-term effects that increase the length of staying in the hospital, cost, morbidity, and mortality. This research aimed to analyze the biofilm formation of S. epidermidis in several types of intravenous fluids based on time. This was a laboratory experimental research using the microtiter plate assay method and crystal violet coloring. Three 96-well microplates were inoculated with S. epidermidis in ringer lactate, 10% dextrose, 5% dextrose, normal saline, and gelafusal, where each plate was incubated at 3 different times of 24, 48, and 72 h, respectively. The results show that the optical density values of all intravenous fluids with bacteria within 24 and 48 h of incubation time did not show any significant differences compared to negative controls, while the 72 h treatments of 10% dextrose, 5% dextrose, and normal saline showed significant differences. This indicates that biofilms of S. epidermidis were not formed in intravenous fluids within 24 and 48 h of incubation time, however this bacterium started forming biofilm in 10% dextrose, 5% dextrose, and normal saline within 72 h of incubation time. In conclusion, the length of incubation time may influence biofilm formation.

483-494

Authors: Azeez Hasan Saleh , Narges Dastmalchi , Parisa Banamolaei , Reza Safaralizadeh

Male infertility can occur due to spermatogenesis defects. The most common reasons for male infertility are azoospermia and oligospermia, which have several underlying factors, one of which is genetic. This study was aimed to investigate the association of azoospermia with the DMRT1 and RBMY1A1 genes polymorphisms and AZFb region microdeletions in Iranian men. Expression of these genes was assessed by RT-PCR. A total of 100 Iranian men with azoospermia, oligozoospermia, or severe oligozoospermia and 100 fertile controls were included in this case-control study. Subjects were genotyped for DMRT1 rs755383 and RBMY1A1 rs1481942953 polymorphisms using Tetra-ARMS PCR. The existence of two sequence-tagged sites (STSs) on the AZFb region was also investigated by multiplex PCR. RT-PCR was used to analyze the expression in the testis of azoospermia cases. With a p-value of 0.038, rs755383 in the DMRT1 gene was related to an elevated risk of azoospermia. However, no significant difference was found in genotype distribution in the RBMY1A1 (rs1481942953) gene polymorphism. Four cases demonstrated Y chromosome microdeletions with sY127 and sY134 markers. Infertile males’ cDNA analysis revealed low expression levels for DMRT1 and PRY (one of the main genes in the AZFb region) with a p-value<0.0001. In contrast, RBMY1A1 expression level did not differ between patients and control groups with a p-value of 0.112. ROC curve analysis was done to detect genes with biomarker potential. With AUCs of 83% and 77%, DMRT1 and PRY had diagnostic marker potential in azoospermia detection.

510-526

Authors: Md. Al Hasibuzzaman , H.M. Iftekhar Alam , Md. Abdur Rashid Mia , Samarth Islam , Sabiha Sultana , Sharmin Ahmed , Afsana Masud , Samiur Rahman , Arysha Alif Khan , Fahmida Hoque Rimti , Ashrak Shad Pyash , Partha Biswas , Homayra Rahman Shoshi , Mahbuba Siddiquy , Fariha Hoque Rimu , Ramiza Nausheen Zaman , Mst. Umme Habiba

Hemoglobin A1C (HBA1c) represents the average serological sugar status of the T2D patients of the past three months, considered a clinically standard method of studying sugar metabolism. Overexpressing HbA1 can metabolically forecast the risk of different cancers among the T2D patients. Based on which, the study aimed to analyze the impact of sugar metabolism in cancer development considering the overexpression of HbA1 as the prognostic biomarker of screening the risks of eight different cancers among the chronic male T2D patients of Bangladesh. Serological analysis of the concentrations of FBS, THABF, creatinine, SC, STGs, HDLC, and LDLC of the T2D patients were conducted in response to their individual HbA1c concentration. Afterward, HbA1 overexpression and promotor-methylation responsible for BLCA, BRCA, CHOL, COAD, LUAD, LUSC, PAAD, and PRAD cancers in the male T2D patients were profiled as the oncoinformatic screening, where the sample types used, individual cancer stages, racial-footprints, gender, age, nodal metastasis, p53-methylations, pancreatitis, diabetes status, smoking behaviors, and survivability status were studied. Finally, the genetic involvement of a group of genes responsible for genetic co-expression of HbA1, endophytic vesicle regulation, antioxidant regulation, and reactive oxygen species based-metabolic regulation in T2D males were identified and comprehensively discussed. The research revealed, significant correlation between BMI and FBS in both the patient and the control groups (p<0.0001). Besides, FBS, THABF, and creatinine were found significantly regulated with their respective HbA1c concentrations (p<0.0001) for each group. The SC, STGs, HDLC, and LDLC regulated ardently and equally for both groups (p<0.0001), while HbA1c ranged from 3.8-5.8% and 5.11-15.8%, for the controls and patients respectively. HbA1 was found interactive with diversified cancer-causing genes, while HbA1 was mostly downregulating with the progressing metastasis. To receive maximum benefits from using HbA1c in clinically profiling of cancer-risks among the chronic-male T2D patients in minimal time and expense further studies can be needed with larger sample size.