American Journal of Molecular Biology

Journal Information
ISSN / EISSN : 2161-6620 / 2161-6663
Published by: Scientific Research Publishing, Inc. (10.4236)
Total articles ≅ 226
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Otilmoi Poul Stephen, Tonny Teya Nyandwaro, Robinson Mugasiali Irekwa, Rebecca Wanjiku Waihenya, Matthew Mutinda Munyao, Peter Kipkemboi Rotich, Caroline Wangui Njoroge, Anne Wanjiru Mwangi, Joanne Jepkemei Yego, Nicole Sian Tanchu, et al.
American Journal of Molecular Biology, Volume 12, pp 43-53; https://doi.org/10.4236/ajmb.2022.122005

Abstract:
Background: A marked decrease in malaria-related deaths worldwide has been attributed to the administration of effective antimalarials against Plasmodium falciparum. However, the continuous spread of P. falciparum resistance to anti-malarial drugs is raising a serious problem in controlling Malaria to the vulnerable children’s immune system. In recent studies, Plasmodium falciparum Kelch 13 propeller gene (Pfk13) has been reported to develop resistance to artemisinin in South Asia. In this study, we checked Plasmodium falciparum chloroquine resistance transporter gene (Pfcrt) involved in chloroquine (CQ) resistance. Method: In this study, archived 280 samples were collected from Alupe primary school children in Busia, Western Kenya from May, 2016 to November, 2016. Genomic DNA was extracted using the MightyPrep reagent. The samples were investigated for P. falciparum positivity out of which 67 of them tested positive giving a prevalence rate of 24%. The sixty-seven were subjected to PCR amplification for the molecular marker resistance to Pfcrt. After PCR amplification, the amplicons were purified and sequenced using Sanger Sequencing. The sequence data were analyzed using BioEdit software to identify point mutations. Results: 14 samples sequences were analyzed on Bioedit software giving the following amino acid changes F76C, Y66H, L70A, Y58C, T59V, V65I, P67L, T81L, Y60S, Y66S, P67T and I71F). New mutations have been reported at position 76 leading to an amino acid change, one of Pfcrt gold standard biomarkers. However, amino acid changes Y66H, L70A, Y58C, T59V, V65I, P67L, T81L, Y60S, Y66S, P67T and I71F are newly reported giving an increase in Pfcrt prevalence of concern from zero to 5.0%. A phylogenetic evolutionary relationship was constructed as shown below. Generally, the results showed a continuous resistance of P.falciparum to Pfcrt which calls for robust continuous monitoring and surveillance. Conclusion: Due to the increase of the resistant Pfcrt gene prevalence, continuous development of new mutants against chloroquine indicates that there is need to repurpose anti-malarial drugs for future partner drugs.
Ning Ma, Xiaohong Li, Renyi Wu
American Journal of Molecular Biology, Volume 12, pp 97-108; https://doi.org/10.4236/ajmb.2022.123009

Abstract:
The aim of the study was to assess the distribution of connexin43 (Cx43) and connexin40 (Cx40) in human and bovine ciliary bodies. The effect of the second messengers cAMP and cyclic cGMP on Cx43 protein expression was also investigated. Enucleated human eyes (remnant after corneal transplantation) and bovine eyes were used. Tissue preparations of the anterior segments of the eyes have proceeded for immunohistochemical staining with polyclonal antibodies of Cx43 and Cx40. Isolated ciliary bodies of human and bovine eyes were incubated with cAMP analog 8-Bromo-cAMP or the cGMP analog 8-Bromo-cGMP, the expression of Cx43 protein in the tissues was then assessed by Western blot assay. Both in human and bovine ciliary bodies, strong immunoreactivity of Cx43, but not Cx40, was observed predominantly in the apical cytoplasmic portions of the pigment ciliary epithelial cells and non-pigmented ciliary epithelial cells. In human ciliary body both cAMP and cGMP up-regulated Cx43 expression, while in the bovine ciliary body, cGMP increased Cx43 expression but cAMP decreased it. Cx43 is the major component of human and bovine gap junctions in the ciliary epithelium. The regulation on the Cx43 expression by cAMP and cGMP in human and bovine ciliary bodies suggests the possibly different roles of these signal messengers in the intracellular communication.
Babacar Faye, Serigne A. A. M. Gueye, Jean A. D. Tine, Hameth Sarr, Alioune Dièye
American Journal of Molecular Biology, Volume 12, pp 54-66; https://doi.org/10.4236/ajmb.2022.122006

Abstract:
Background: Different studies have demonstrated high prevalence of HPV infection and dysplastic lesions of the cervix in immunocompromised patient such as women living with HIV. Is this high prevalence due to a greater susceptibility to HPV infection, which is known to be frequent in its latent form in women? Objective: This study aims to identify HPV genotypes in HIV+ and HIV− women to understand HPV molecular epidemiology in Senegal. Material and Method: Endocervical samples from 331 HIV+ and HIV− women, sexually active, were collected. The molecular identification of the 28 genotypes studied (19 HPV-HR and 9 HPV-LR) was carried out after DNA extraction, by multiplex PCR with the Anyplex™ II HPV28 detection kit from Seegene on CFX96™ Bio-Rad machine. The comparisons were made by calculating the p-value and odds ratio with R Studio software (version 4.1.0). The results were considered significant if p − women. Conclusion: Our results showed that the prevalence of HPV, HPV-HR and HPV-BR was significantly higher in HIV+ women. Non-vaccine genotypes were among the most found genotypes. Groups of HIV+ women aged between 35 and 50, married and using contraception were significantly more infected with HPV than the same groups of HIV-women.
Sanaa Mohammed Yousif, Adam Dawoud Abakar, Salaheldein Gumaa Elzaki, Salma Omer Ibrahim, Omer Abu Elhasan, Mohamed Taj-Eldin, Elhadi Abdalla Ahmed
American Journal of Molecular Biology, Volume 12, pp 30-42; https://doi.org/10.4236/ajmb.2022.122004

Abstract:
Background: The characteristics of Staphylococcus aureus that made it the most important cause of wound infections are environmental spread antimicrobials resistance and virulence. Absence of molecular detection of drug resistance and virulence factors in many developing countries limits the epidemiological information. This study conducted to identify PVL virulence gene, and blaOXA-23 and blaOXA-51 drug resistance genes of Staphylococcus aureus isolated from surgical-sites infections (SSIs) and traumatic wounds. Methods: A cross-sectional study was conducted from 2019 to 2021, in which 70 cefepime resistant Staphylococcus aureus were used, the strains were isolated from patients of SSIs and traumatic wounds admitted to the department of General Surgery in Wad Medani Teaching Hospital. Mannitol salt agar was used for primary culture followed by biochemical identification and Kirby Bauer susceptibility testing. Single and multiplex PCR protocols performed for bacterial confirmation and target genes detection. Results: Staphylococcus aureus strains from SSIs constituted 56% (39/70) from which 41% (16/39) possessed PVL gene while 42% (13/31) of wound infections strains were positive for PVL gene. Presence of PVL gene was significantly associated with resistance to meropenem (P. value 0.023) and ceftriaxone (P. value 0.037). blaOXA-23 was significantly detected with resistance to meropenem, augmentin and ceftriaxone. While blaOXA-51 was significantly identified among Staphylococcus aureus strains that showed resistance to meropenem and ciprofloxacin. Conclusion: This is the first study in Sudan that identified blaOXA-23 and blaOXA-51 in Staphylococcus aureus and correlated them to resistance to commonly used antimicrobials. Meropenem resistant Staphylococcus aureus were significantly positive for PVL, blaOXA-23 and baOXA-51 genes.
Ying Liu, Yanan Jiang, Chao Wang, Haiying Zhang, Yan Liu
American Journal of Molecular Biology, Volume 12, pp 11-22; https://doi.org/10.4236/ajmb.2022.122002

Abstract:
Idiopathic pulmonary fibrosis is an untreatable lethal lung disease, which is related to the aberrant proliferation of fibroblasts. M3 muscarinic acetylcholine receptor (M3-mAChR) activation exerts proliferative effect on various kinds of cells. However, whether M3-mAChR inhibition has a protective effect on pulmonary fibrosis remains unexplored. A rat model of pulmonary fibrosis was established by intratracheal instillation of bleomycin. Darifenacin was used to block M3-mAChR. Histological changes were observed using Masson’s Trichrome and hematoxylin and eosin (HE) staining. Hydroxyproline was measured by Hydroxyproline detection kit. Transforming growth factor β1 (TGF-β1) and tumor necrosis factor-α (TNF-α) were measured by enzyme-linked immunosorbent assay (ELISA). In vitro, pulmonary fibroblasts were isolated from lungs of neonatal rat. After treatment, the cell viability, Hydroxyproline level was measured by MTT and Hydroxyproline detection kit respectively. The expression level of extracellular signal-regulated kinase (ERK), nuclear factor kappa-B (N-NF-κB), and microRNA-21 (miR-21) was detected by western blot or quantitative real-time PCR (qRT-PCR). Darifenacin relieved the fibrotic effects provoked by bleomycin. The expression level of hydroxyproline, TGF-β1 and TNF-α level was all downregulated after darifenacin treatment. In lung fibroblasts, darifenacin decreased cell viability and hydroxyproline level induced by bleomycin. Besides, phosphorylation-ERK and nuclear N-NF-κB protein level was downregulated, as well as miR-21 level. M3-mAChR antagonist darifenacin attenuates bleomycin-induced pulmonary fibrosis in rats, which may relate to the ERK/NF-κB/miRNA-21 signaling pathway.
Huan Zhao, Xiaoyu Jiang, Lingyu Lu, Qing Lin
American Journal of Molecular Biology, Volume 12, pp 1-9; https://doi.org/10.4236/ajmb.2022.121001

Abstract:
Acute respiratory infection in children (ARTI) is the most common childhood infectious disease, and its pathogens include bacteria, fungi, viruses, chlamydia, mycoplasma and rickettsia. In recent years, with the continuous development of pathogen detection methods, the diagnosis and treatment of acute respiratory infections has received more and more attention from clinicians. The clinical diagnosis and treatment of acute respiratory infections in children and the research of laboratory detection methods have also been continuously developed. The manuscript presents a review of progress in the clinical diagnosis, treatment and laboratory testing of acute respiratory infections in children by collecting references.
Usenobong F. Ufot, Monday I. Akpanabiatu, Khasim Cali, Ifiok D. Uffia, Inyang Udosen
American Journal of Molecular Biology, Volume 12, pp 67-84; https://doi.org/10.4236/ajmb.2022.122007

Abstract:
The goal of this study was to determine whether mutation of the Mn-binding site of wild-type recombinant Phlebia radiata manganese peroxidase 3 affected the pH-dependence kinetic parameters. pH range investigated was 2.5 – 12.0. The catalytic efficiency of the mutant enzymes at high and low pH in comparison to the wild-type was investigated using standard rPr-MnP3 protocol. Wild-type recombinant Phlebia radiata MnP3 enzyme showed optimal activity with Mn (II) as substrate at pH 5.0 and remained moderately active (approximately 40%) in the pH range of 6.0 - 9.0. The rPr-MnP3 mutants’ maximum activity ranged between 5.5 and 8.0. Wild-type and mutants rPr-MnP3 enzymes exhibited a similar pH profile with optimum pH of 3.0 for ABTS oxidation. Mutation has severely decreased the catalytic efficiency for Mn (II) oxidation at pH 5.0. The rPr-MnP3 enzymes showed enhanced affinity for Mn (II) at alkaline pH and a more alkaline range for catalysis than ever reported for any Manganese Peroxidase. This study reveals that at higher pH, rPr-MnP3 can function with alternative ligands in the Mn (II) site and does not have an absolutely obligate requirement for an all carboxylate ligand set. These results further strongly confirm that Mn2+ binding site is the only productive catalytic site for Mn (II) oxidation.
Keita Odanaka
American Journal of Molecular Biology, Volume 12, pp 23-29; https://doi.org/10.4236/ajmb.2022.122003

Abstract:
Candida albicans proliferates in the skin and oral cavity and is the causative agent of candida dermatitis and oral candidiasis. C. albicans is known to form biofilms on oral mucosa and denture surfaces. Formation of biofilms deteriorates the permeability of antifungal drugs, decreasing their effectiveness. Therefore, in this study, I identified a compound with inhibitory activity against C. albicans biofilm formation. Heat shock protein 90 was selected as the target protein, and a potential ligand for the same was extracted and identified as 2-(4-methylpiperazin-1-yl)cyclopentanol. C. albicans was then cultured with varying concentrations of this compound: 0 mmol/L, 0.63 mmol/l. 2.5 mmol/l, and 10 mmol/l, and biofilm formation was measured via crystal violet assay. The findings demonstrated that 2-(4-methylpiperazin-1-yl)cyclopentanol substantially inhibits biofilm formation when added at a concentration of 0.63 mmol/l or higher. It is suggested that C. albicans could be eliminated more efficiently using this compound in combination with the existing antifungal drug miconazole. Further, the compound may also be useful as a disinfectant for medical devices, such as catheters, to prevent the formation of C. albicans biofilms.
Salma Omer Ibrahim, Elimam M. A. Mohammed, Sami Mahjoub Taha, Sanaa Mohammed Yousif, Hajir Omer, Omer Omer, Mirghani Seif-Elnasr, Seitelbanat Yassin, Yousif Abdelhameed Mohammed, Omer Abu Elhasan, et al.
American Journal of Molecular Biology, Volume 12, pp 85-96; https://doi.org/10.4236/ajmb.2022.123008

Abstract:
Background: Urosepsis is one of the most common infections that require empirical broad spectrum antibiotics immediately after diagnosis. This has led to development of bacterial resistance by acquiring the capability to destroy the β-lactam ring. Methodology: This is a cross-sectional hospital-based study. The study was conducted from 2019 to 2020 at Gezira Hospital for Renal diseases and surgery (GHRDS). A hundred patients were diagnosed clinically with urosepsis and the isolated organisms were Escherichia coli, Staphylococcus aureus, Proteus mirabilis, Klebsiella pneumonia and Pseudomonas aeruginosa. The susceptibility test was conducted by Kirby Bauer disc diffusion technique according to clinical laboratory standard institute (CLSI) guidelines. Seventy eight samples of bacterial genomic DNA were confirmed by 16srRNA and multiplex PCR, were performed for genotypic blaOXA-51 and blaOXA-23 gene characterization of isolated bacteria. Then gel electrophoresis was used to identify the presence or absence of (blaOXA-51 and blaOXA-23) genes. Results: 88.5% (69/78) in 16srRNA detected. Using multiplex PCR, the frequencies of blaOXA-51 and blaOXA-23 genes were 13% and 10.1%, respectively. The percentages of isolates which yielded both blaOXA-51 and blaOXA-23 among P. aeruginosa was 25% (1/4), among K. pneumonia was 17% (1/6), and among E. coli was 8% (3/37). Only blaOXA-51 was detected in P. mirabilis 10% (1/10) and only blaOXA-23 was detected in S. aureus 5% (1/18). Conclusion: In this study, the presence of blaOXA-51 and blaOXA-23 genes was increased in the isolated bacteria.
Aiju Zhang, Zhiming Zhou
American Journal of Molecular Biology, Volume 11, pp 38-50; https://doi.org/10.4236/ajmb.2021.112004

Abstract:
Plasma Membrane Calcium ATPase (PMCA) plays a critical role in transporting Ca2+ out of the cytosol across the plasma membrane. Here, a full-length cDNA sequence of plasma membrane Ca2+-ATPase gene was isolated from the gill of Hyriopsis cumingii (HcPMCA) by using SMART RACE technique. The entire cDNA was 5230 bp, including a 417-bp 5'-UTR, a 3588-bp ORF and a 1225-bp 3'-UTR, encoding a 1195-amino acid protein, and no putative signal peptide was predicted. Compared with PMCA homologs from seawater mollusks, HcPMCA had high similarity with them in both sequence and structure. Tissue-specific expression analysis revealed that HcPMCA mRNA was detected in all the sampled tissues, but was prominently expressed in the gill and mantle. When exposed to a serie of increasing Ca2+ that lasted for 7 days, the mRNA expression of HcPMCA in the mantle was slightly downregulated, but peaked at 60 mg/L. Moreover, the temporal expression of HcPMCA transcripts in the mantle after 60 mg/L Ca2+ exposure was shown to be bell-shaped, which was slightly downregulated at 24 h, but upregulated from 24 h to 48 h post-treatment, peaking at 48 h. The result of present study provides useful information for further studies on function and regulation mechanism of HcPMCA gene.
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