Impact of COVID-19 infections and vaccination on menstrual cycle symptoms in the south of Jordan: a cross-sectional study.

BACKGROUND: Several recent studies have highlighted the need for more evaluation of the impact of COVID-19 infections and vaccines on the reproductive system and menstruation. This study aimed to assess the impact of COVID-19 infection and vaccines on menstrual symptoms. METHODS: A cross-sectional survey utilizing face-to-face interviews from January 1 to 31 March 2022 was conducted in the city of Al-Karak in southern Jordan. The questionnaire included sociodemographic characteristics, medical and reproductive history, the contraceptive method used if any, menstrual cycle (MC) status, previous medical and drug history, and the impact of infection and vaccination on the MC. RESULTS: The study questionnaire was completed by 400 participants with a mean age of 32.1±12.6 years. Regarding the history of COVID-19 infections, 33.8% of the participants reported a history of confirmed COVID-19 infections, 77.8% of them did not report any menstrual changes following the infection, while the remaining 22.2% reported changes in menstruation. The most commonly reported post-COVID-19 manifestations were irregular (27.6%) and light menstrual cycle (MC) (24.15) or dysmenorrhea (24.1%). Heavy menstruation was reported by 17.2% of participants post-COVID-19 infection. Two-thirds of the study participants (66.6%) reported no changes in the MC following the administration of the COVID-19 vaccine. The most reported symptoms for those who experienced changes in the MC following the vaccination were irregular cycle (13.1%), heavy menstruation (7%), and light menstruation (7%). Other reported symptoms were dysmenorrhea (4.6%), intermenstrual bleeding (1.2%), and amenorrhea (0.5%). CONCLUSION: This study revealed minor changes in the MC following COVID-19 infections and administration of the COVID-19 vaccine. These findings are consistent with published reports. It is recommended that future clinical trials for new vaccines for women of childbearing age include outcomes related to sex hormones and MC. Women should be encouraged to take the vaccines and report symptoms to healthcare professionals for further assessment.

Emergence and Genomic Characterization of a spa Type t4407 ST6-SCCmec Type IVa Methicillin-Resistant Staphylococcus aureus Strain Isolated from Al-Karak Hospital, Jordan.

Background and Objectives: Methicillin-resistant Staphylococcus aureus (MRSA) is a major concern in Jordanian hospitals in terms of infection control. The purpose of this study was to identify the resistance patterns of Staphylococcus aureus strains isolated from surfaces of critical locations within the Al-Karak Governmental Hospital in 2019. Additionally, the study aimed to conduct whole-genome sequencing on the isolates. Materials and Methods: In February 2019, fourteen S. aureus strains were isolated from surfaces in critical sites in the Al-Karak Governmental Hospital. These isolates underwent antibiogram testing to determine their resistance profile. Genome sequencing using the Illumina MiSeq platform was applied to the extracted DNA from these isolates. The genomic data, including coding sequences, were analyzed to identify lineage, resistance genes, and plasmids. Results: The antibiogram results revealed that 11 of the 14 isolates were resistant to oxacillin, 6 to linezolid, and 1 to rifampicin, while none showed resistance to chloramphenicol. Eleven isolates were identified as MRSA, with a novel spa type (t4407) not previously reported in Jordan. High-quality sequencing data were obtained for only one isolate, i.e., A29, the genome showed 2,789,641 bp with a 32.7% GC content and contained 2650 coding sequences. Genomic analysis indicated the ST6 lineage, mecA gene (SCCmec type IVa(2B)), and a hybrid plasmid (pJOR_blaZ) carrying the blaZ gene for ß-lactam resistance. Genomic data were deposited in NCBI (CP104989). The A29 genome closely resembled an MRSA genome isolated from a Danish hospital in 2011. The SNP analysis revealed identical antimicrobial resistance genes in these two genomes. Conclusions: This study unveils the first genomic sequence of an MRSA isolate from Jordan, marked by distinctive genotypic traits. The findings enhance our understanding of the MRSA types circulating in Jordan and the region and substantiate the phenomenon of intercontinental MRSA transmission.

COVID-19 pandemic impact on antibiotics sensitivity of E. coli and K. pneumoniae from urine specimens: a retrospective study.

INTRODUCTION: Escherichia coli and Klebsiella pneumoniae are common organisms associated with urinary tract infections. The COVID-19 pandemic has had a negative impact on antibiotics misuse globally. This study analyzed the antibiotic susceptibility for these two pathogens isolated from urine samples during the period of 18 months before and after the COVID-19 pandemic. METHODOLOGY: This retrospective study was conducted in Al-Karak government referral and teaching hospital in Jordan. The study included two groups; group A included urine samples from September 2018 to March 11, 2020, while group B from March 12, 2020 to August 2021. Samples were analyzed using the automated VITEK 2 system and the analysis of results was done using the WHONET version 5.6. RESULTS: A total of 642 E. coli and 113 K. pneumoniae were isolated and analyzed. The antibiogram showed a significant overall increase in antibiotic susceptibility of both bacteria during the pandemic period (group B). The sensitivity has significantly increased by 75% (15/20) and 50% (10/20) for all antibiotics used for E. coli and K. pneumoniae respectively. On the other hand, E. coli showed a significant increase in resistance to ceftriaxone (13.4%) and gentamicin (6.4%). A similar trend of an increase in resistance to gentamicin (17.4%) was also noticed among K. pneumoniae isolates. CONCLUSIONS: The antimicrobial susceptibility pattern for urine isolates showed an increased overall sensitivity and an increased resistance to ceftriaxone and gentamicin during the pandemic period. Our results highlight the need for revising and updating the antimicrobial stewardship programs post-COVID pandemic utilizing local data.

Deregulation of the cyclin-dependent kinase inhibitor p27 as a putative candidate for transformation in Chlamydia trachomatis infected mesenchymal stem cells.

Purpose: Several pathological conditions might cause the degradation of the cyclin-dependent kinase inhibitor (CKI) p27 and cell cycle arrest at the G1 phase, including cancers and infections. Chlamydia trachomatis (Ctr), as an obligatory intracellular pathogen, has been found to alter the fate of the cell from different aspects. In this study, we aimed to investigate the effect of Ctr infection on the expression of the important cell cycle regularity protein p27 in mesenchymal stem cells (MSCs). Methods: Isolation of MSCs from healthy human fallopian tube was confirmed by detection of the stemness markers Sox2, Nanog and Oct4 and the surface markers CD44, CD73 and CD90 by Western blotting and fluorescence-activated cell sorting analysis. The expression of p27 was downregulated at the protein level upon Ctr D infection measured by Real-Time Quantitative Reverse Transcription PCR (qRT-PCR), IF and Western blotting. Recovery of p27 in Ctr D-infected MSCs was achieved by treatment with difluoromethylornithine (DFMO). Ctr D infected MSCs were able to produce colonies in anchorage-independent soft agar assay. Conclusion: Ctr D infection was able to downregulate the expression of the important cell cycle regulator protein p27, which will be considered a putative candidate for transformation in Ctr D infected MSCs.

Diabetic foot ulcer in Southern Jordan: A cross-sectional Study of Clinical and Microbiological Aspects.

Background: Diabetes mellitus (DM) is a common metabolic disorder that significantly affects public health. Diabetic foot ulcer (DFU) is one of the serious complications of diabetes. DFU has a wide spectrum of bacterial isolates comprising Gram-positive, Gram-negative, aerobic bacteria and anaerobes. In the last two decades there has been an increase in the multidrug-resistant isolates (MDR). Materials and methods: This cross-sectional prospective observational study was conducted in southern Jordan among patients with DFU. The included variables are sociodemographic and clinical information. Isolates from swab culture of ulcers and antimicrobial susceptibility pattern are also recorded. Results: A total of 64 diabetic patients with DFU were included in this study. Most patients included in the study were males with male-to-female ratio of (2.2:1). The mean age was 54 years (SD ± 10.7). The mean duration of DM was 16.4 years (SD ± 7.5) and the mean HbA1c was 9.9% (SD ± 2.1). Neuropathy and anemia were noted in 72% and 44% of patients, respectively. The most frequent bacterial isolates were gram negative bacteria accounts for 29 isolates (45.3%). About 37.5% (24) of bacterial isolates showed MDR pattern. Previous antibiotic use in the last 30 days showed significant association with MDR bacteria (p-value <0.05). Previous history of amputations, presence of neuropathy, renal impairment, retinopathy, presence of anemia, limited joint mobility and presence of foot deformity were significantly associated with Wagner's grade ≥ three. Conclusion: Many factors affect and increase the risk of having high grade diabetic foot ulcer. The most frequent bacterial isolates from diabetic foot ulcers were gram negative bacteria. High rates of MDR in this study reflect the loose implementation of regulations in Jordan regarding antibiotics dispensing.

Loss of p53 Expression in Gastric Epithelial Cells of Helicobacter pylori-Infected Jordanian Patients.

Background: Around half of the global population is chronically infected with the stomach bacterium Helicobacter pylori, making it one of the most common chronic infections worldwide. H. pylori induces the production of reactive oxygen species, DNA damage, and accelerates the degradation of the tumor suppressor protein p53, which may lead to cancer development. In this study, we investigated the relationship between H. pylori infection and the expression of p53 in gastric mucosa in a group of patients from Jordan. Methods: In this retrospective case-control study, the epithelium of gastric glands in subjects chronically infected with H. pylori was examined for the expression of p53. Paraffin-embedded gastric biopsy samples from the archives for 50 Jordanian patients diagnosed with chronic H. pylori infection and 25 samples free of H. pylori infection and any other gastric abnormalities were selected. Samples were analyzed for the presence of H. pylori as well as p53 expression levels in the mucosa and submucosa by immunohistochemical analyses and Western blotting. Results: H. pylori was detected in the gastric tissues of infected individuals (n = 50); whereas, no H. pylori infection was detected in uninfected healthy individuals (n = 25) using immunohistochemistry. In contrast to the noninfected samples of gastric mucosa, no nuclear p53 expression was detected in the infected samples using immunohistochemistry. In addition, the levels of p53 in H. pylori-positive samples detected by Western blotting were significantly lower than those in the negative individuals. Conclusion: Our data reveal that p53 protein expression decreased in gastric mucosa of patients infected with H. pylori. The loss of this tumor suppressor may play a role in the increased risk for tumor initiation associated with H. pylori carriage.

Humoral immune response to COVID-19 infection or vaccination among celiac disease patients.

Introduction: Celiac disease (CD) is the most common autoimmune disease (AD) of the small intestine, affecting 1-2% of the population globally. It is characterized by the serological presence of autoantibodies (Abs), tissue transglutaminase antibody (tTGA), immunoglobulin (Ig) A, and IgG. Production of antibodies against SARS-CoV-2 after infection with the virus or vaccination is not well understood, especially among CD patients. The goal of this study was to measure the IgG antibodies in Jordanian patients infected with or vaccinated against the SARS-CoV-2 virus with different types of vaccines (Pfizer- BioNTech BNT162b2, Sinopharm BBIBP-CorV or Oxford-AstraZeneca ChAdOx1-S) and compare them with the levels in non-celiac controls. IgG levels induced by different vaccines were also compared. Material and methods: The data for this cross-sectional study were obtained via a survey, whereby respondents were identified through convenience sampling. The healthy controls were given Questionnaire A while CD patients completed Questionnaire B. The blood samples from all participants were tested for the COVID-19 nucleocapsid protein (NP) IgG serum levels for participants previously infected with SARS-CoV-2, and spike (S) protein (S1/S2) IgG serum levels for vaccine recipients. Results: The study involved 116 individuals, 60 (51.7%) of whom were CD patients. The NP IgG serum levels in the infected and S1/S2 IgG levels in the vaccinated CD patients were significantly lower than the levels in controls (48.3 ±44.5 vs. 81.1 ±34.4 and 49 ±45.8 vs. 75.7 ±38.6, p = 0.002). Moreover, only the Pfizer vaccine induced significantly more IgG antibodies in controls compared to CD patients (88.8 ±29.1 vs. 58.3 ±45.4, p = 0.01). On the other hand, the IgG levels were significantly higher in CD patients who received the Pfizer relative to the AstraZeneca vaccine (58.3 ±45.5 vs. 13.0 ±23.6, p = 0.03). After adjusting for presence of CD, age, sex, body mass index (BMI), comorbidities, vaccine type, smoking, gluten adherence, and time since infection or vaccination, SARS-CoV-2 S1/S2 IgG Abs and/or NP IgG Abs positivity was significantly associated with CD absence and negatively with vaccine type (AstraZeneca) with the odds ratios (ORs) of 9.6 (95% CI = 1.5-59.2, p = 0.015) and 0.03 (95% CI = 0.004-0.244. p = 0.001), respectively. Conclusions: We concluded that patients with CD had lower SARS-CoV-2 S1/S2 IgG Abs and NP IgG Abs levels than controls, and CD patients who received the Pfizer vaccine had higher IgG levels than patients who received the AstraZeneca vaccine. We recommend that further research be conducted to address the dynamics of the antibody responses in CD patients regarding COVID-19 infection.

The Use of 13C-Urea Breath Test for Non-Invasive Diagnosis of Helicobacter pylori Infection in Comparison to Endoscopy and Stool Antigen Test.

Helicobacter pylori (H. pylori) can cause gastritis, peptic ulcer diseases and gastric carcinoma. Endoscopy as the gold standard method of diagnosis is an invasive procedure that might not be suitable in all scenarios. Therefore, this first study in Jordan aimed to assess the non-invasive 13C urea breath test (UBT) and stool antigen test for diagnosis of H. pylori infection and the successfulness of eradication therapy as alternatives for endoscopy. Hence, a total of 30 patients attending the endoscopy units at Alkarak teaching hospital were asked to complete a questionnaire with demographic and clinical data. They were then tested for H. pylori using 13C UBT and H. pylori stool antigen before having endoscopy. Another 30 patients who were positive for H. pylori by endoscopy were tested using both tests 6 weeks post eradication therapy. Results showed that the rate of H. pylori detection using endoscopy was 56.7% (17/30). Heartburns (82.3%, p value = 0.019), epigastric pain (88.2%, p value = 0.007) and vomiting (70.5%, p value = 0.02) were the most significant symptoms. Family history of peptic ulcer diseases was significantly associated with an increased risk for having a H. pylori positive result (p value = 0.02). Compared to endoscopy, the sensitivity of 13C UBT for the diagnosis of H. pylori was 94.1% (16/17), while it was 76.5% (13/17) for the stool antigen test. The specificity of both tests was equal (76.9%). However, the positive predictive and negative predictive values (84.2% and 90.9%) for 13C UBT were higher than those (81.3% and 71.4%) for the stool antigen test. The accuracy of 13C UBT was 86.7% compared to 76.7% for the stool antigen test. There was an 87% agreement (20 patients out of 23) between both tests when used to assess success of the eradication therapy. In conclusion, the 13C UBT was found to be more sensitive and accurate than the stool antigen test when used for diagnosis; furthermore, it has a comparable outcome to the stool antigen test in assessing the successfulness of the eradication treatment.

Human papillomavirus as an independent risk factor of invasive cervical and endometrial carcinomas in Jordan.

BACKGROUND: Endometrial and cervical carcinomas are the most common gynecologic malignancies in Western world and many countries. The human papillomavirus (HPV) high-risk genotypes are associated with cervical carcinoma (CC). Chlamydia trachomatis (C. trachomatis), the most common sexually transmitted bacterial infection worldwide, considered a cofactor for HPV infection and CC. Information on HPV infection rate and type distribution among Jordanian women having CC is currently limited and unavailable among those with endometrial carcinoma. Therefore, the present study aimed to provide an updated estimate on HPV infection rate and its high-risk genotypes' distribution among Jordanian women by comparing data from invasive cervical carcinoma (ICC) to normal cervical tissues. Similarly, assessment of HPV infection rate was extended to the endometrial tissues. C. trachomatis infection was investigated as well to explore its possibility as HPV cofactor for induction of such carcinomas. METHODS: Total DNA was extracted from 144 formaldehyde-fixed paraffin-embedded cervical and endometrial tissue, equally divided between age-matched control and carcinoma cases. Polymerase chain reaction (PCR) was used for general detection of HPV-DNA, high risk HPV-16 and 18 genotypes and C. trachomatis DNA using specific primers. RESULTS: HPV infection was detected in 91.7% and 61.1% of cervical cancer patients and controls, respectively. Likewise, it was higher among cases (47.2%) than controls (13.8%) in endometrial biopsies. Significantly higher HPV infection rates were found among ICC and endometrial control biopsies of women >50 years. Out of 33 HPV positive ICC cases, single HPV-16 infections were detected in 69.7% compared to HPV-18 (15.2%), while HPV-16/18 co-infections were only found in three (9%) samples. C. trachomatis was not detected in all studied groups. CONCLUSION: The present study has successfully provided an updated estimate on HPV infection rate among Jordanian women with and without ICC and endometrial carcinoma. In addition, a lack of co-infection was observed between HPV and C. trachomatis in both cancer types.

Analysis of Helicobacter pylori Genotypes Amongst Jordanians' Dental Plaque Samples.

BACKGROUND: Helicobacter pylori (H. pylori) infection has been associated with gastritis, gastric ulcer, mucosa-associated lymphoid tissue lymphoma and gastric cancer. The prevalence of H. pylori virulence genes have been studied in different populations and from different sources of samples but their prevalence has not been studied in dental plaque in Jordanian people; therefore, the aim of this study was to determine the genotypes of H. pylori isolated from dental plaque samples. METHODS: Dental plaque samples were collected from 60 Jordanian volunteers. The genotypes of H. pylori virulence genes including the cytotoxin-associated gene A (cagA) and the vacuolating toxin (vacA) were determined using polymerase chain reaction (PCR). RESULTS: The cagA gene was detected in 14 (23.3%) samples, while vacA was detected in all volunteers enrolled in this study (100%). The most prevalent vacA alleles were m2 and s1 in 54 (90%) and 55 (91.7%) of volunteers, respectively. Compared to the other combinations including the most virulent vacA genotype s1/m1 which was detected in 11 (18.2%) of volunteers, the most prevalent vacA allelic combinations were s1/m2 and s2/m2 in 56 (93.3%) and 27 (45%) of volunteers, respectively. CONCLUSIONS: These results indicate a significant carriage of virulent H. pylori strains among Jordanian people in their dental plaques, which increases the possible transmission of these strains among them. In addition, the studying of the genotypic pattern of H. pylori virulence genes in the dental plaque could represent an essential tool for infection prevention and predicting the severity and prognosis of H. pylori gastric infection.

Chlamydia trachomatis Prevents Apoptosis Via Activation of PDPK1-MYC and Enhanced Mitochondrial Binding of Hexokinase II.

The intracellular human bacterial pathogen Chlamydia trachomatis pursues effective strategies to protect infected cells against death-inducing stimuli. Here, we show that Chlamydia trachomatis infection evokes 3-phosphoinositide-dependent protein kinase-1 (PDPK1) signaling to ensure the completion of its developmental cycle, further leading to the phosphorylation and stabilization of MYC. Using biochemical approaches and imaging we demonstrate that Chlamydia-induced PDPK1-MYC signaling induces host hexokinase II (HKII), which becomes enriched and translocated to the mitochondria. Strikingly, preventing the HKII interaction with mitochondria using exogenous peptides triggers apoptosis of infected cells as does inhibiting either PDPK1 or MYC, which also disrupts intracellular development of Chlamydia trachomatis. These findings identify a previously unknown pathway activated by Chlamydia infection, which exhibits pro-carcinogenic features. Targeting the PDPK1-MYC-HKII-axis may provide a strategy to overcome therapeutic resistance of infection.

Chlamydia infection depends on a functional MDM2-p53 axis.

Chlamydia, a major human bacterial pathogen, assumes effective strategies to protect infected cells against death-inducing stimuli, thereby ensuring completion of its developmental cycle. Paired with its capacity to cause extensive host DNA damage, this poses a potential risk of malignant transformation, consistent with circumstantial epidemiological evidence. Here we reveal a dramatic depletion of p53, a tumor suppressor deregulated in many cancers, during Chlamydia infection. Using biochemical approaches and live imaging of individual cells, we demonstrate that p53 diminution requires phosphorylation of Murine Double Minute 2 (MDM2; a ubiquitin ligase) and subsequent interaction of phospho-MDM2 with p53 before induced proteasomal degradation. Strikingly, inhibition of the p53-MDM2 interaction is sufficient to disrupt intracellular development of Chlamydia and interferes with the pathogen's anti-apoptotic effect on host cells. This highlights the dependency of the pathogen on a functional MDM2-p53 axis and lends support to a potentially pro-carcinogenic effect of chlamydial infection.

Chlamydia trachomatis remodels stable microtubules to coordinate Golgi stack recruitment to the chlamydial inclusion surface.

Chlamydia trachomatis (Ctr), an obligate intracellular bacterium, survives and replicates within a membrane-bound vacuole, termed the inclusion, which intercepts host exocytic pathways to acquire nutrients. Ctr subverts cellular trafficking pathways from the Golgi by targeting small GTPases, including Rab proteins, to sustain intracellular bacterial replication; however, the precise mechanisms involved remain incompletely understood. Here, we show that Chlamydia infection in human epithelial cells induces microtubule remodeling, in particular the formation of detyrosinated stable MTs, to recruit Golgi ministacks, but not recycling endosomes, to the inclusion. These stable microtubules show increased resistance to chemically induced depolymerization, and their selective depletion results in reduced bacterial infectivity. Rab6 knockdown reversibly prevented not only Golgi ministack formation but also detyrosinated microtubule association with the inclusion. Our data demonstrate that Chlamydia co-opts the function of stable microtubules to support its development.

Chlamydia trachomatis inhibits inducible NO synthase in human mesenchymal stem cells by stimulating polyamine synthesis.

Chlamydia trachomatis is considered the most common agent of sexually transmitted disease worldwide. As an obligate intracellular bacterium, it relies on the host for survival. Production of NO is an effective antimicrobial defense mechanism of the innate immune system. However, whether NO is able to arrest chlamydial growth remains unclear. Similarly, little is known about the mechanisms underlying subversion of cellular innate immunity by C. trachomatis. By analyzing protein and mRNA expression in infected human mesenchymal stem cells, combined with RNA interference and biochemical assays, we observed that infection with C. trachomatis led to downregulated expression of inducible NO synthase (iNOS) in human mesenchymal stem cells in vitro. Furthermore, infection upregulated the expression of the rate-limiting enzyme in the polyamine biosynthetic pathway, ornithine decarboxylase, diverting the iNOS substrate l-arginine toward the synthesis of polyamines. Inhibition of ornithine decarboxylase activity using small interfering RNA or the competitive inhibitor difluoromethylornithine restored iNOS protein expression and activity in infected cells and inhibited chlamydial growth. This inhibition was mediated through tyrosine nitration of chlamydial protein by peroxynitrite, an NO metabolite. Thus, Chlamydia evades innate immunity by inhibiting NO production through induction of the alternative polyamine pathway.

Autophagy restricts Chlamydia trachomatis growth in human macrophages via IFNG-inducible guanylate binding proteins.

Interferon γ (IFNG) is a key host response regulator of intracellular pathogen replication, including that of Chlamydia spp The antichlamydial functions of IFNG manifest in a strictly host, cell-type and chlamydial strain dependent manner. It has been recently shown that the IFNG-inducible family of immunity-related GTPases (IRG) proteins plays a key role in the defense against nonhost adapted chlamydia strains in murine epithelial cells. In humans, IFN-inducible guanylate binding proteins (hGBPs) have been shown to potentiate the antichlamydial effect of IFNG; however, how hGBPs regulate this property of IFNG is unknown. In this study, we identified hGBP1/2 as important resistance factors against C. trachomatis infection in IFNG-stimulated human macrophages. Exogenous IFNG reduced chlamydial infectivity by 50 percent in wild-type cells, whereas shRNA hGBP1/2 knockdown macrophages fully supported chlamydial growth in the presence of exogenous IFNG. hGBP1/2 were recruited to bacterial inclusions in human macrophages upon stimulation with IFNG, which triggered rerouting of the typically nonfusogenic bacterial inclusions for lysosomal degradation. Inhibition of lysosomal activity and autophagy impaired the IFNG-mediated elimination of inclusions. Thus, hGBP1/2 are critical effectors of antichlamydial IFNG responses in human macrophages. Through their capacity to remodel classically nonfusogenic chlamydial inclusions and stimulate fusion with autophagosomes, hGBP1/2 disable a major chlamydial virulence mechanism and contribute to IFNG-mediated pathogen clearance.