The Prevalence of Extended Spectrum Beta-Lactamase Genes SHV, TEM and CTX-M in Clinical Isolates of Proteus mirabilis Ramina Khoshaba Iraqi Journal of Science, 2025 This study investigates the antibiotic resistance patterns in Proteus mirabilis isolates, with a specific focus on the presence of Beta-Lactamase genes, including SHV, TEM, and CTX-M, which are known to contribute to the bacterium's ability to cause various diseases. A total of 100 urine samples were collected from patients confirmed urinary tract infections (UTIs), and out of these, only 20 of those samples were positively identified as Proteus mirabilis using cultural characteristics, biochemical examinations, and the Vitek II system. Antibiotic susceptibility was tested for 27 antibiotics. Results showed that isolates exhibited multi-drug resistance ranging from 10 to 19 antibiotics, with 100% resistance to Ampicillin, Erythromycin, Clindamycin, Clarithromycin, Penicillin G, Cephalothin and Cefaclor. Additionally, the isolates revealed high resistance to the third generation Cephalosporines of, including Ceftriaxone (85%), Cefotaxime (75%), Ceftazidime (55%), and Cefoperazone (45%). Using the Double Disk Synergy Test (DDST), eight isolates were confirmed to produce ESβL. Conventional PCR was conducted with specific primers to detect four distinct ESβLs (TEM, SHV, CTX-M-8, and CTX-M-9). Results revealed that CTX-M-8 was found in all eight isolates with phenotypic evidence of ESβL production, indicating that they are the most prevalent type of ESβL. At the same time, those for CTX-M-9, TEM-type and SHV type were 87.5%, 62.5% and 12.5 % respectively. In conclusion, all the examined isolates were found to produce ESβLs, particularly the CTX-M-8 gene.
Antibiotic Resistance, Biofilm Formation, and Identification of FimH and FimA Adhesion Genes in Uropathogenic Escherichia Coli (UPEC) Isolated from Patients in Baghdad Province Safa A. Abdul Hamid, Ramina M. Khoshabeh Iraqi Journal of Science, 2024 Urinary tract infections (UTIs) are one of the most common infectious disorders worldwide. The most frequent cause of UTIs is uropathogenic Escherichia coli (UPEC). The current study is designed to assess the resistance of antibiotics, the formation of biofilm, and the detection of adhesion genes in E. coli isolated from patients with UTIs. A hundred and fifty samples were collected from patients with confirmed UTIs in Baghdad province during the period of October 2022 to February 2023. Isolation and identification of E. coli were performed using cultural characteristics, gram staining, and biochemical examination, and the results indicated that 52 (35%) isolates were identified as E. coli. Eleven antibiotic discs were utilized to estimate the sensitivity of E. coli isolates. 98% of isolates were resistant to ampicillin, followed by 75.5% of isolates resistant to both trimethoprim/sulfamethoxazole and fosfomycin, followed by 73.5, 71.4, 67.3, 53.1, and 6.1% of isolates resistant to imipenem, ceftazidime, nitrofurantoin, cefepime, and amikacin. All isolates were sensitive to piperacillin-tazobactam, ciprofloxacin, and aztreonam at 100%, 100%, and 98%, respectively. The microtiter plate method was utilized to estimate the ability of E. coli to form biofilm. The majority of isolates (69.2%; n = 36) were moderate biofilm producers, while 19.2% (n = 10) and 11.5% (n = 6) of isolates were strong and weak biofilm producers, respectively. Polymerase Chain Reactions (PCR) were applied to determine the gene expression level of adhesion genes (fimH and fimA genes). Forty isolates harbored both fimH and fimA genes.
AKR1B8 deficiency drives severe DSS-induced acute colitis through invasion of luminal bacteria and activation of innate immunity Qiulin Deng, Yichen Yao, Jing Yang, Ramina Khoshaba, Yi Shen, et al. Frontiers in Immunology, 2022 BackgroundDysfunction of intestinal epithelial cells (IECs) promotes inflammatory bowel disease (IBD) and associated colorectal cancer (CRC). AKR1B8 deficiency impairs the IEC barrier function, leading to susceptibility to chronic colitis induced by dextran sulfate sodium (DSS), yet it remains unclear how acute colitic response is in AKR1B8 deficient mice.MethodsAKR1B8 knockout (KO) and littermate wild type mice were exposed to oral 1.5% DSS in drinking water for 6 days. Disease activity index and histopathological inflammation scores by H&E staining were calculated for colitic severity; permeability was assessed by fluorescein isothiocyanate dextran (FITC-Dextran) probes and bacterial invasion and transmission were detected by in situ hybridization in mucosa or by culture in blood agar plates. Immunofluorescent staining and flow cytometry were applied for immune cell quantification. Toll-like receptor 4 (TLR4) and target gene expression was analyzed by Western blotting and qRT-PCR.ResultsAKR1B8 KO mice developed severe acute colitis at a low dose (1.5%) of DSS in drinking water compared to wild type controls. In AKR1B8 KO mice, FITC-dextran was penetrated easily and luminal bacteria invaded to the surface of IEC layer on day 3, and excessive bacteria translocated into the colonic mucosa, mesenteric lymph nodes (MLNs) and liver on day 6, which was much mild in wild type mice. Hyper-infiltration of neutrophils and basophils occurred in AKR1B8 KO mice, and monocytes in spleen and macrophages in colonic mucosa increased markedly compared to wild type mice. TLR4 signaling in colonic epithelial cells of AKR1B8 KO mice was activated to promote great IL-1β and IL-6 expression compared to wild type mice.ConclusionsAKR1B8 deficiency in IECs drives severe acute colitis induced by DSS at a low dose through activation of the innate immunity, being a novel pathogenic factor of colitis.
Impaired Barrier Function and Immunity in the Colon of Aldo-Keto Reductase 1B8 Deficient Mice Xin Wang, Ramina Khoshaba, Yi Shen, Yu Cao, Minglin Lin, et al. Frontiers in Cell and Developmental Biology, 2021 Aldo-keto reductase 1B10 (AKR1B10) is downregulated in human ulcerative colitis (UC) and colorectal cancer, being a potential pathogenic factor of these diseases. Aldo-keto reductase 1B8 (AKR1B8) is the ortholog in mice of human AKR1B10. Targeted AKR1B8 deficiency disrupts homeostasis of epithelial self-renewal and leads to susceptibility to colitis and carcinogenesis. In this study, we found that in AKR1B8 deficient mice, Muc2 expression in colon was diminished, and permeability of colonic epithelium increased. Within 24 h, orally administered FITC-dextran penetrated into mesenteric lymph nodes (MLN) and liver in AKR1B8 deficient mice, but not in wild type controls. In the colon of AKR1B8 deficient mice, neutrophils and mast cells were markedly infiltrated, γδT cells were numerically and functionally impaired, and dendritic cell development was altered. Furthermore, Th1, Th2, and Th17 cells decreased, but Treg and CD8T cells increased in the colon and MLN of AKR1B8 deficient mice. In colonic epithelial cells of AKR1B8 deficient mice, p-AKT (T308 and S473), p-ERK1/2, p-IKBα, p-p65 (S536), and IKKα expression decreased, accompanied with downregulation of IL18 and CCL20 and upregulation of IL1β and CCL8. These data suggest AKR1B8 deficiency leads to abnormalities of intestinal epithelial barrier and immunity in colon.
Curcumin nicotinate selectively induces cancer cell apoptosis and cycle arrest through a P53-mediated mechanism Ying-chun He, Lan He, Ramina Khoshaba, Fang-guo Lu, Chuan Cai, et al. Molecules, 2019 Curcumin is an anticancer agent, but adverse effects and low bioavailability are its main drawbacks, which drives efforts in chemical modifications of curcumin. This study evaluated antiproliferative activity and cancer cell selectivity of a curcumin derivative, curcumin nicotinate (CN), in which two niacin molecules were introduced. Our data showed that CN effectively inhibited proliferation and clonogenic growth of colon (HCT116), breast (MCF-7) and nasopharyngeal (CNE2, 5-8F and 6-10B) cancer cells with IC50 at 27.7 μM, 73.4 μM, 64.7 μM, 46.3 μM, and 31.2 μM, respectively. In cancer cells, CN induced apoptosis and cell cycle arrest at G2/M phase through a p53-mediated mechanism, where p53 was activated, p21 and pro-apoptotic proteins Bid and Bak were upregulated, and PARP was cleaved. In non-transformed human mammary epithelial cells MCF10A, CN at 50 µM had no cytotoxicity and p53 was not activated, but curcumin at 12.5 µM activated p53 and p21 and inhibited MCF10A cell growth. These data suggest that CN inhibits cell growth and proliferation through p53-mediated apoptosis and cell cycle arrest with cancer cell selectivity.
Long non-coding RNA UASR1 promotes proliferation and migration of breast cancer cells through the AKT/mTOR pathway Zhe Cao, Ping Wu, Min Su, Hongyan Ling, Ramina Khoshaba, et al. Journal of Cancer, 2019 Long non-coding RNAs (lncRNAs) are non-coding RNAs longer than 200 nucleotides that function as regulatory factors in many human diseases, including cancer. However, majority of lncRNAs remain to be characterized. In this study, we characterized a novel lncRNA transcript, named UNC5B antisense RNA1 (UASR1). UASR1 is 647bp in length consisting of two exons. This lncRNA is an antisense of intron 1 of unc-5 netrin receptor B (UNC5B) gene. In breast cancer tissues, UASR1 was upregulated. Ectopic expression of UASR1 promoted proliferation and clonogenic growth of breast cancer cells MCF7 and MDA-MB-231. The migration of these cells also increased as demonstrated by wound healing and transwell assays. In contrast, silencing of UASR1 suppressed cell proliferation and migration. Further studies showed that UASR1 activated AKT and AKT-mediated mTOR signaling pathway to stimulate cell proliferation and growth. In these cells, active pAKT, pTSC2, p4EBP1 and pp70S6K were increased. Taken together, our data suggest that UASR1 plays an oncogenic role in breast cancer cells through activation of the AKT/mTOR signaling pathway, being a novel RNA oncogene.
AKR1B10 activates diacylglycerol (DAG) second messenger in breast cancer cells Chenfei Huang, Zhe Cao, Jun Ma, Yi Shen, Yiwen Bu, et al. Molecular Carcinogenesis, 2018 Aldo‐keto reductase 1B10 (AKR1B10) is upregulated in breast cancer and promotes tumor growth and metastasis. However, little is known of the molecular mechanisms of action. Herein we report that AKR1B10 activates lipid second messengers to stimulate cell proliferation. Our data showed that ectopic expression of AKR1B10 in breast cancer cells MCF‐7 promoted lipogenesis and enhanced levels of lipid second messengers, including phosphatidylinositol bisphosphate (PIP2), diacylglycerol (DAG), and inositol triphosphate (IP3). In contrast, silencing of AKR1B10 in breast cancer cells BT‐20 and colon cancer cells HCT‐8 led to decrease of these lipid messengers. Qualitative analyses by liquid chromatography‐mass spectrum (LC‐MS) revealed that AKR1B10 regulated the cellular levels of total DAG and majority of subspecies. This in turn modulated the phosphorylation of protein kinase C (PKC) isoforms PKCδ (Thr505), PKCµ (Ser744/748), and PKCα/βII (Thr638/641) and activity of the PKC‐mediated c‐Raf/MEK/ERK signaling cascade. A pan inhibitor of PKC (Go6983) blocked ERK1/2 activation by AKR1B10. In these cells, phospho‐p90RSK, phospho‐MSK, and Cyclin D1 expression was increased by AKR1B10, and pharmacological inhibition of the ERK signaling cascade with MEK1/2 inhibitors U0126 and PD98059 eradicated induction of phospho‐p90RSK, phospho‐MSK, and Cyclin D1. In breast cancer cells, AKR1B10 promoted the clonogenic growth and proliferation of breast cancer cells in two‐dimension (2D) and three‐dimension (3D) cultures and tumor growth in immunodeficient female nude mice through activation of the PKC/ERK pathway. These data suggest that AKR1B10 stimulates breast cancer cell growth and proliferation through activation of DAG‐mediated PKC/ERK signaling pathway.
