Association of Human Papilloma Virus, Cytomegalovirus, and Epstein-Barr Virus with Breast Cancer in Jordanian Women.

Background and Objectives: The investigation of oncogenic viruses and their potential association with breast cancer (BC) remains an intriguing area of study. The current work aims to assess evidence of three specific viruses, human papillomavirus (HPV), cytomegalovirus (CMV), and Epstein-Barr virus (EBV) in BC samples and to explore their relationship with relevant clinicopathological variables. Materials and Methods: The analysis involved BC samples from 110 Jordanian female patients diagnosed with BC and breast tissue samples from 30 control patients with no evidence of breast malignancy, investigated using real-time PCR. The findings were then correlated with various clinico-pathological characteristics of BC. Results: HPV was detected in 27 (24.5%), CMV in 15 (13.6%), and EBV in 18 (16.4%) BC patients. None of the control samples was positive for HPV or CMV while EBV was detected in only one (3.3%) sample. While (HPV/EBV), (HPV/CMV), and (EBV/CMV) co-infections were reported in 1.8%, 2.7%, and 5.5%, respectively, coinfection with the three viruses (HPV/CMV/EBV) was not reported in our cohort. A statistically significant association was observed between HPV status and age (p = 0.047), and between clinical stage and CMV infection (p = 0.015). Conclusions: Our findings indicate the presence or co-presence of HPV, CMV, and EBV in the BC subpopulation, suggesting a potential role in its development and/or progression. Further investigation is required to elucidate the underlying mechanisms that account for the exact role of oncoviruses in breast carcinogenesis.

Synthesis, docking study, and antitumor evaluation of benzamides and oxadiazole derivatives of 3-phenoxybenzoic acid as VEGFR-2 inhibitors.

Current chemotherapeutic agents have several limitations, including lack of selectivity, the development of undesirable side effects, and chemoresistance. As a result, there is an unmet need for the development of novel small molecules with minimal side effects and the ability to specifically target tumor cells. A new series of 3-phenoxybenzoic acid derivatives, including 1,3,4-oxadiazole derivatives (4a-d) and benzamides derivatives (5a-e) were synthesized; their chemical structures were confirmed by Fourier-transform infrared spectroscopy, 1H nuclear magnetic resonance (NMR), 13C NMR, and mass spectra; and various physicochemical properties were determined. The antiproliferative activities of the new derivatives were evaluated by means of the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Three compounds (4b, 4c, and 4d) exhibited cytotoxicity against two of the three cell lines tested, five compounds (3, 4a, 5a, 5b, and 5e) were toxic to one cell line, while two compounds (5c and 5d) were not cytotoxic to any of the three cell lines tested in the current study. Based on docking scores, MTT assay findings, and vascular endothelial growth factor receptor 2 (VEGFR-2) kinase activity data, Compound 4d was selected for further biological investigation. Flow cytometry was used to determine the mode of cell death (apoptosis vs. necrosis) and the effect on cell cycle progression. Compound 4d arrested HepG2 hepatocellular carcinoma cells in the G2/M phase and activated both the intrinsic and extrinsic apoptosis pathways. In conclusion, Compound 4d has shown promising results for future research as a potent VEGFR-2 tyrosine kinase inhibitor.

Antimicrobial Resistance Molecular Mechanisms of Helicobacter pylori in Jordanian Children: A Cross-Sectional Observational Study.

BACKGROUND: H. pylori antimicrobial resistance causes increasing treatment failure rates among H. pylori gastritis in children. This study investigates the molecular mechanisms of H. pylori antimicrobial resistance among Jordanian children. METHODS: Demographic, clinical, and laboratory data were recorded for children referred to Prince Hamzah Hospital. Clarithromycin, Metronidazole, and Levofloxacin susceptibility were tested via E-test. Clarithromycin-related mutations were investigated using Real-Time (RT)-PCR and Levofloxacin resistance was analyzed with DNA sequencing of the gyrA gene. RESULTS: 116 children were recruited, including 55.2% females and 55.2% in the age range of 10.1 to 14 years. A total of 82.7% were naïve to eradication therapy. H. pylori positivity was 93.9%, 89.6%, 61.7%, and 84.3% according to Rapid Urease Test, histology, culture, and RT-PCR, respectively. Resistance rates were 25.9% for Clarithromycin, 50% for Metronidazole, and 6.9% for Levofloxacin via E-test. A2142G or A2143G or a combination of both mutations concerning Clarithromycin resistance were documented in 26.1% of samples, while mutations in gyrA gen-related to Levofloxacin resistance were reported in 5.3% of samples. Antibiotic resistance was significantly affected by abdominal pain, anemia, hematemesis, and histological findings (p < 0.05). CONCLUSION: H. pylori resistance was documented for Metronidazole and Clarithromycin. RT-PCR for H. pylori identification and microbial resistance determination are valuable alternatives for cultures in determining antimicrobial susceptibility.

Prevalence of SARS-COV-2 and other respiratory pathogens among a Jordanian subpopulation during Delta-to-Omicron transition: Winter 2021/2022.

Acute respiratory tract infections (ARTIs) during the winter months are associated with higher morbidity and mortality compared to other seasons of the year, with children below five, elderly, and immunocompromised patients being the most susceptible. Influenza A and B viruses, rhinovirus, coronaviruses, respiratory syncytial virus, adenovirus, and parainfluenza viruses, are the most frequently identified causes of viral ARTIs. In addition, the emergence of SARS-CoV-2 in 2019 provided an additional viral cause of ARTIs. The aim of this study was to provide an overview of the epidemiological status of upper respiratory infections, their main causative agents, and reported clinical presentation in the winter months of 2021, during two important surges of COVID-19 in Jordan. Nasopharyngeal samples were collected from 339 symptomatic patients during the period from December 2021 to March 2022, followed by nucleic acid isolation using a Viral RNA/DNA extraction Kit. The causative virus species associated with the patient's respiratory symptoms was determined utilizing a multiplex real-time PCR targeting 21 viruses, 11 bacteria, and a single fungus. SARS-CoV-2 was identified in 39.2% of the patients (n = 133/339). A total of 15 different pathogens were also identified as co-infections among these 133 patients (n = 67/133). SARS-CoV-2-Bacterial coinfections (37.6%, n = 50/133) were the most frequent, with Bordetella species being the most common, followed by Staphylococcus aureus, and H.influenzae type B. Viral coinfection rate was 27.8% (n = 37/133), with Influenza B virus and Human bocavirus being the most common. In Conclusion, Both SARS-CoV-2, influenza B virus, and Bordetella accounted for the majority of infections in patients with URTI during the winter months of 2021-2022. Interestingly, more than 50% of the patients with symptoms of URTIs were confirmed to have a coinfection with two or more respiratory pathogens, with SARS-CoV-2 and Bordetella coinfection being most predominant.

Senolytic Therapy: A Potential Approach for the Elimination of Oncogene-Induced Senescent HPV-Positive Cells.

Senescence represents a unique cellular stress response characterized by a stable growth arrest, macromolecular alterations, and wide spectrum changes in gene expression. Classically, senescence is the end-product of progressive telomeric attrition resulting from the repetitive division of somatic cells. In addition, senescent cells accumulate in premalignant lesions, in part, as a product of oncogene hyperactivation, reflecting one element of the tumor suppressive function of senescence. Oncogenic processes that induce senescence include overexpression/hyperactivation of H-Ras, B-Raf, and cyclin E as well as inactivation of PTEN. Oncogenic viruses, such as Human Papilloma Virus (HPV), have also been shown to induce senescence. High-risk strains of HPV drive the immortalization, and hence transformation, of cervical epithelial cells via several mechanisms, but primarily via deregulation of the cell cycle, and possibly, by facilitating escape from senescence. Despite the wide and successful utilization of HPV vaccines in reducing the incidence of cervical cancer, this measure is not effective in preventing cancer development in individuals already positive for HPV. Accordingly, in this commentary, we focus on the potential contribution of oncogene and HPV-induced senescence (OIS) in cervical cancer. We further consider the potential utility of senolytic agents for the elimination of HPV-harboring senescent cells as a strategy for reducing HPV-driven transformation and the risk of cervical cancer development.

Phenotypic and Molecular Screening of Nasal S. aureus from Adult Hospitalized Patients for Methicillin- and Vancomycin-resistance.

INTRODUCTION: Nasal carriers of Staphylococcus aureus are common and play an important role in the transmission of infections. The aim of this study is a phenotypic and molecular investigation of nasal methicillin- and vancomycin-resistant S. aureus in hospitalized patients. METHODS: 202 nasal swabs were collected from patients at Prince Hamzah Hospital, Jordan, through 2016-2017. Swabs were processed according to standard microbiological procedures to isolate Staphylococci. Antibiotic susceptibility testing was performed using disk diffusion, E-test, microdilution and Vitek 2. Methicillin resistance was confirmed by testing for the mecA gene, while vancomycin resistance was screened by testing for the vanA and vanB genes. RESULTS: The mean age of participants was 50.17±18.18 years and 59.4% were females. Nasal Staphylococci was isolated in 64/202 (31.7%), S. aureus was isolated from 33 samples (16.3%), MRSA was isolated from 13 samples (6.4%) and constitutive Macrolide-lincosamidestreptogramin B (MLSB) was isolated from 12 samples (5.9%). All MRSA isolates harbored the mecA gene. All isolates were sensitive to vancomycin using E-test and the microdilution test and were negative for the vanA and vanB genes. The highest resistance rate was observed for benzylpenicillin (>90%), while the lowest resistance rate was for tobramycin (<5%) among all isolates. Nasal Staphylococci, S. aureus and MRSA colonization significantly correlate with increased number of family members and previous hospitalization (P<0.05), while nasal S. aureus significantly correlates with a history of skin infection (P=0.003). CONCLUSION: Nasal colonization by mecA-mediated MRSA is common among hospitalized patients, while vanA- and vanB-mediated vancomycin resistance was not detected in any nasal isolates.

Status of Biofilm-Forming Genes among Jordanian Nasal Carriers of Methicillin-Sensitive and Methicillin-Resistant Staphylococcus aureus.

Background: Biofilm formation in Staphylococcus aureus is a major virulence factor. Both methicillin-sensitive Staphylococcus aureus (MSSA) and methicillin-resistant Staphylococcus aureus (MRSA) are common causes of community- and hospital-acquired infections and are associated with biofilm formation. The status of biofilm-forming genes has not been explored in Jordanian nasal carriers of S. aureus. This study investigates antibiotic resistance patterns and the prevalence of biofilm-forming genes between MSSA and MRSA in two distinct populations in Jordan. Methods: A total of 35 MSSA and 22 MRSA isolates were recovered from hospitalized patients and medical students at Prince Hamzah Hospital, Jordan. Antibiotic susceptibility was tested using disk diffusion method and Vitek 2 system. The phenotypic biofilm formation was tested using Congo red agar and microtiter plate assays. The prevalence of the biofilm-forming genes was determined using multiplex PCR. Results: Among 57 S. aureus isolates, 22 (38.6%) isolates were MRSA and were highly resistant against benzylpenicillin, oxacillin, and imipenem. The frequencies of the icaADBC were 77.1%, 97.1%, 94.3%, and 97.1% respectively in MSSA compared to 86.4%, 100%, 100%, and 100% in MRSA isolates. On the other hand, the frequency of the fnbA, fnbB, clfA, fib, clfB, ebps, eno, and cna genes was 81.8%, 90.9%, 95.5%, 90.9%, 86.4%, 100%, 100%, and 40.9%, respectively in the MRSA isolates. Conclusion: In both groups, MRSA isolates, in comparison to MSSA, were significantly more resistant to cefoxitin, oxacillin, imipenem, tetracycline, clindamycin, and trimethoprim-sulfamethoxazole. Unexpectedly, biofilm formation and gene prevalence between MRSA and MSSA isolates showed no significant difference, suggesting other potential virulence mechanisms.

Methicillin and vancomycin resistance in coagulase-negative Staphylococci isolated from the nostrils of hospitalized patients.

INTRODUCTION: Nasal colonization by coagulase-negative Staphylococci (CoNS) play an important role in nosocomial infections. This study aims to determine antibiotics susceptibility pattern and molecular screening of methicillin- and vancomycin-resistant nasal CoNS among hospitalized patients. METHODOLOGY: Nasal swabs were collected from 202 inpatients at Prince Hamzah Hospital, Jordan. Swabs were processed according to standard microbiological procedures to isolate Staphylococci. Antibiotic susceptibility testing was performed using disk diffusion, E-test, microdilution, and Vitek 2. Molecular analysis was performed using PCR for the detection of mecA, vanA, and vanB genes. RESULTS: Nasal Staphylococci was isolated in 64/202 (31.7%) samples. Thirty isolates (14.8%) were CoNS, including S. haemolyticus (n = 17, 8.4%), S. sciuri (n = 6, 3%), S. epidermidis (n = 2, 1%), S. warneri (n = 2, 1%), S. hominis (n = 2, 1%), and S. lentus (n = 1, 0.5%). Twenty-two (10.9%) isolates were MR-CoNS harboring mecA gene. CoNS and MR-CoNS isolates were highly resistant to benzylpenicillin, erythromycin, fosfomycin, and imipenem. All isolates were sensitive to vancomycin by E-test and microdilution test and were negative for vanA and vanB genes. Nasal CoNS colonization was associated with an increased number of family members living with the participant (P = 0.04) and with admission to the orthopedic department (P = 0.03), while MR-CoNS colonization was associated with smoking (P = 0.03). CONCLUSIONS: Nasal colonization by unusual CoNS species and mecA-positive MR-CoNS are common among hospitalized patients. Absence of vanA and vanB genes suggests little contribution of nasal CoNS to vancomycin resistance transmission.

Seroprevalence of cystic echinococcosis in a high-risk area (Al-Mafraq Governorate) in Jordan, using indirect hemagglutination test.

Hydatid disease (HD) is a zoonotic disease of humans and animals which is caused by infection with the larval stages of the taeniid cestodes of the genus Echinococcus. HD is endemic in many countries of the Middle East, including Jordan. The seroprevalence rate of HD in areas of elevated risk in Jordan has not previously been investigated using indirect haemagglutination (IHA) testing. In the present study, 512 blood samples were collected from recruited outpatients from an internal medicine clinic in Al-Mafraq Governmental Hospital in Jordan. Each participant signed a consent form and completed a questionnaire. The presence of antibodies specific for E. granulosus antigens was detected using an IHA test. Statistical analysis was performed by SPSS software using the Chi-square test. In all, 4.1% of the study participants were seropositive for E. granulosus IgG antibodies. There was a significant correlation between unexplained weight loss among seropositive patients (P = 0.018). Seropositivity was significantly higher in patients who slaughtered sheep inside their houses (P = 0.023). HD seroprevalence did not correlate with gender (P = 0.433), age (P = 0.880), residency status (P = 0.938), or educational level (P = 0.808). The vast majority (75.2%) of participants reported no prior knowledge about HD, and 99.8% were not aware about the etiology of the disease.

Isolation of Fully Vancomycin-Resistant Streptococcus thoraltensis from the Nasal Cavity of a Healthy Young Adult.

BACKGROUND: Streptococcus thoraltensis was first isolated from pigs and rabbits. Later, isolation from human oral and nasal cavities and from throat and oropharynx was documented. S. thoraltensis was isolated from patients with periodontitis, tonsillopharyngitis, and chorioamnionitis suggesting a possible pathological role in human infections. All S. thoraltensis isolates of animal and human origins were sensitive to vancomycin. METHODS: Standard microbiological identification methods, biochemical analysis, and antibiotic susceptibility testing using disk diffusion and E methods were used. Automatic species identification and antibiotic susceptibility testing were carried out using the Vitek 2 compact system. Molecular analysis of vancomycin resistance gene was carried out using a PCR with specific primers for vanA. RESULTS: We report a healthy young female adult, aged 19 years, with history of exposure to pet rabbit who had nasal colonization with S. thoraltensis. Identification of S. thoraltensis was based on traditional microbiological methods (culture, Gram stain, and biochemical tests), and the Vitek 2 compact system with 97% confidence rate. Antibiotic susceptibility testing of the isolate indicated resistance to most antibiotics, including penicillins, cephalosporins, methicillin, and glycopeptides. The minimal inhibitory concentration for vancomycin and teicoplanin was exceptionally high (>256 µg/mL). Molecular analysis indicated the absence of vanA gene in S. thoraltensis. CONCLUSION: We report for the first time the isolation of a fully vancomycin-resistant S. thoraltensis independent of vanA from a healthy human anterior nasal cavity. The pathological role of this newly identified organism with an exceptionally rare resistance pattern in human infections is yet to be identified.

Nasal colonization by methicillin-sensitive and methicillin-resistant Staphylococcus aureus among medical students.

INTRODUCTION: Nasal carriers of methicillin-resistant Staphylococcus aureus (MRSA) are common and play an important role in nosocomial infections. The prevalence rate and characterization of nasal carriers of MRSA among medical students in Jordan has not been investigated before. METHODOLOGY: The resistance of S. aureus to several antibiotics was tested using disc diffusion method, automatic Vitek 2, and penicillin binding protein (PBP) 2 slide test. Bacterial species and resistance genes were confirmed using molecular analysis of three relevant genes by real-time PCR. Two hundred ninety nasal swabs were collected from medical students at Hashemite University from June 2015 to August 2016. All participants signed a voluntary consent form and filled a predesigned questionnaire. RESULTS: The mean age of participants was 19.7 ± 2 years and 61.7% of them were males. 63 out of the 290 (21.7%) samples were identified to have S. aureus, 56 (19.3%) were methicillin-sensitive S. aureus (MSSA) and 7 (2.4%) were MRSA. S. aureus nasal colonization significantly associates with male gender (OR = 1.7, CI = 0.94-3.18, P = 0.049) and chronic illnesses (OR = 4.0, CI = 1.52-10.65, P = 0.006). Consistency between disc diffusion, Vitek 2, and PBP 2 methods for MRSA screening were satisfactory compared to molecular analysis. All MRSA samples were positive for SCCmec:orfx junction gene (MRSA-specific), nuc gene (S. aureus- specific), mecA gene (PBP-mediated resistant), and PBP2 production. All MRSA isolates were multi-drug resistant and were sensitive to Linezolid, Vancomycin, and Tigecycline. CONCLUSIONS: This study confirms that nasal colonization by MRSA among medical students necessitates further attention to prevent nosocomial infections.