Cefixime and cefixime-clavulanate for screening and confirmation of extended-spectrum beta-lactamases in Escherichia coli.

INTRODUCTION: Detection of Extended-Spectrum Beta-Lactamases (ESBLs) depends on screening for resistance to certain cephalosporins, confirmation with selective ESBL inhibitors, and ESBL genes detection. New tests are required for accurate ESBL detection. AIMS: To test the ability of cefixime (CFM) and cefixime-amoxicillin/clavulanate (CFM-AMC) as a screening and confirmatory test for ESBL identification. METHODS: 246 clinical isolates of Escherichia coli were tested by an ESBL screening test, a double-disk synergy test (DDST), a disk replacement test, the Vitek 2 ESBL test, and an ESBL genes test by PCR. CFM ESBL Screening was performed by disk diffusion, while CFM-AMC confirmation was performed by DDST and a disk replacement test. RESULTS: 246 E. coli clinical isolates from two referral hospitals were collected over 2 years. The mean age ± standard deviation of patients was 43.8 ± 27.7 years and 76.8% were females. Resistance rates to penicillins, first, second, and third generation cephalosporins, and monobactams were very high at 97%, 84%, 100% and 97%, respectively. ESBL screening was positive in 81.3% of isolates, DDST was positive in 74.8%, disk replacement was positive in 79%, Vitek 2 ESBL test was positive in 67.3%, and ESBL genes were detected in 85.8% of isolates (CTX-M 75%, TEM 42.5%, SHV 4.6%). Compared to genotyping, screening with CFM achieved 87.7% sensitivity and 64.7% specificity. CFM-AMC DDST achieved 75.8% sensitivity and 75.4% specificity, and CFM-AMC disk replacement had 73% sensitivity and 70% specificity. CONCLUSIONS: High prevalence of ESBLs was noted among E. coli isolates, dominated by CTX-M genotype. ESBL screening and confirmation using CFM and CFM-AMC is a new and accurate method for ESBLs detection.

The utility of platelet activation biomarkers in thrombotic microangiopathies.

Primary thrombotic microangiopathies (TMAs) are observed in thrombotic thrombocytopenic purpura (TTP) and hemolytic uremic syndrome (HUS), while secondary TMAs have a wide range of etiologies. Early diagnosis and treatment of TMA are critical for patient well-being; however, distinguishing TTP from HUS on presentation is particularly challenging. Thrombocytopenia and platelet activation are central to different types of TMAs, thus limiting the utility of standard diagnostic approaches to evaluate the platelet function and hemostatic capacity. Alternative means of quantifying and monitoring changes to platelet activation and function are urgently needed. Activated platelets have been shown to interact with proteins of the complement and coagulation cascades and form part of inflammation processes engaged in TMA. Increased levels of platelet surface receptors as well as increased plasma levels of platelet-derived soluble proteins have been reported in TMAs. Elevated levels of platelet-leukocyte aggregates and platelet microparticles are also reported in different types of TMAs. Larger prospective evaluations of platelet activation markers in TMA using standardized assays, with comparison to cohorts of patients with thrombosis, coagulopathy, and thrombocytopenia, to evaluate the clinical usefulness of platelet markers in TMA are now needed. This review will summarize the current knowledge around platelet activation markers and critically evaluate their utility in diagnosis and prognosis of TMA patients.

The platelet Fc receptor, FcγRIIa.

Human platelets express FcγRIIa, the low-affinity receptor for the constant fragment (Fc) of immunoglobulin (Ig) G that is also found on neutrophils, monocytes, and macrophages. Engagement of this receptor on platelets by immune complexes triggers intracellular signaling events that lead to platelet activation and aggregation. Importantly these events occur in vivo, particularly in response to pathological immune complexes, and engagement of this receptor on platelets has been causally linked to disease pathology. In this review, we will highlight some of the key features of this receptor in the context of the platelet surface, and examine the functions of platelet FcγRIIa in normal hemostasis and in response to injury and infection. This review will also highlight pathological consequences of engagement of this receptor in platelet-based autoimmune disorders. Finally, we present some new data investigating whether levels of the extracellular ligand-binding region of platelet glycoprotein VI which is rapidly shed upon engagement of platelet FcγRIIa by autoantibodies, can report on the presence of pathological anti-heparin/platelet factor 4 immune complexes and thus identify patients with pathological autoantibodies who are at the greatest risk of developing life-threatening thrombosis in the setting of heparin-induced thrombocytopenia.

Pathologic shear triggers shedding of vascular receptors: a novel mechanism for down-regulation of platelet glycoprotein VI in stenosed coronary vessels.

Ligand-induced ectodomain shedding of glycoprotein VI (GPVI) is a metalloproteinase-dependent event. We examined whether shear force, in the absence of GPVI ligand, was sufficient to induce shedding of GPVI. Human-citrated platelet-rich plasma or washed platelets were subjected to increasing shear rates in a cone-plate viscometer, and levels of intact and cleaved GPVI were examined by Western blot and ELISA. Pathophysiologic shear rates (3000-10 000 seconds(-1)) induced platelet aggregation and metalloproteinase-dependent appearance of soluble GPVI ectodomain, and GPVI platelet remnant. Shedding of GPVI continued after transient exposure to shear. Blockade of α(IIb)ß(3), GPIbα, or intracellular signaling inhibited shear-induced platelet aggregation but minimally affected shear-induced shedding of GPVI. Shear-induced GPVI shedding also occurred in platelet-rich plasma or washed platelets isolated from a von Willebrand disease type 3 patient with no detectable VWF, implying that shear-induced activation of platelet metalloproteinases can occur in the absence of GPVI and GPIbα ligands. Significantly elevated levels of sGPVI were observed in 10 patients with stable angina pectoris, with well-defined single vessel coronary artery disease and mean intracoronary shear estimates at 2935 seconds(-1) (peak shear, 19 224 seconds(-1)). Loss of GPVI in platelets exposed to shear has potential implications for the stability of a forming thrombus at arterial shear rates.

The Impact of COVID-19 on Mental Health and Sleep Function of Hospital Staff Working With COVID-19 Patients: A Cross-Sectional Nationwide Study.

The coronavirus disease 2019 (COVID-19) pandemic has presented a globally challenging situation for human physical and mental health. Healthcare workers (HCWs) are affected by increased levels of anxiety, stress, and insomnia. This study aimed to evaluate the effect of COVID-19 on HCWs anxiety, stress, and insomnia levels. This cross-sectional study employed the Hospital Anxiety and Depression Scale, Perceived Stress Scale 10, and Insomnia Severity Index to assess anxiety, stress, and insomnia among HCWs at 10 COVID-19 isolation and treatment hospitals/centers after the first COVID-19 wave in Jordan. A web-based survey was used to collect data from 183 participants. Statistical analysis of factors affecting the mean scores of anxiety, stress, and insomnia was carried using student t-test or ANOVA while factors associated with differences in anxiety, stress, and insomnia frequencies were tested using Chi-square/Fisher exact test. Multivariate analysis was performed to determine the independent risk factors. Among participants, 97.3% reported moderate to severe levels of stress, 68% reported borderline to high abnormal levels of anxiety, and 32% had moderate to severe insomnia. The mean of anxiety total score was 9.8 ± 4.8, stress total score was 22.7 ± 4.5, and insomnia total score was 11.0 ± 7.1. Significant positive correlations were noted between anxiety, stress, and insomnia (P < .005). Female gender, migraine, less working years, increased time spent with patients, lower workforce, clinical insomnia and high stress were significant independent factors associated with anxiety (P < .05). Younger age, being single or divorced, heart disease, smoking, occupation (nurses), lower workforce, vaccination dose, and anxiety were significant independent factors associated with insomnia (P < .05). Increased time spent with patients, lower workforce, lower spouse and colleagues support, sadness due to isolation and anxiety were significant independent factors associated with stress. HCWs at COVID-19 centers had high levels of stress, anxiety, and insomnia. Appropriate interventions to maintain HCWs mental health are recommended.

Quantitative and equipment-free paper-based agglutination assay of bacterial cells.

Background: point-of-care (POC) tests are useful for bedside/home applications, emergencies, frequent follow-ups, and resource-limited areas. Limited quantitative and equipment-free POC assays have been reported. This study aims to develop, validate, and apply a simple, quantitative, paper-based POC assay. Methods: wax-channeled paper treated with specific anti-Brucella and anti-Salmonella antibodies was used for distance-based chromatographic elution of stained bacterial cell agglutinations. Results: a qualitative paper-based agglutination POC test was developed using color intensity, tail appearance, and "+/-" signs that clearly distinguish the positive and negative results. The optimization of the test for paper type, microfluidic channel design, antibody and bacterial cell concentrations, and elution methods was carried out. Quantitative assay transformation was successfully developed using the color intensity of the original reaction zone, intensity of elution tail, and distance-based migration that correspond to bacterial agglutination size. The migration distance of eluted bacterial agglutination bands corresponds to the target concentration with good linearity and minimal variability. Reporting of colored band migration with numbers using microfluidic patterns was used to enhance non-technical end-user applications. A distance-based POC assay prototype was then successfully used for the accurate detection of known and unknown samples in comparison with standard assays. Conclusions: the migration distance of an eluted stained bacterial agglutination correlated with anti-bacterial antibody concentrations. A simple, cheap, quantitative, and equipment-free paper-based POC assay of bacterial cell agglutination was developed. This test can be used for simple "+/-" results, thermometer-like quantification, or text reporting with numbers corresponding to target concentrations. The assay has extended applications to different human disease biomarkers.

Multi-locus sequence typing of Escherichia coli isolated from clinical samples in Jordan.

INTRODUCTION: Escherichia coli (E. coli) is the major cause of extraintestinal infections in the urinary tracts and bloodstream in humans in the community and health care institutions. Several studies on the genetic characterization of E. coli among clinical and environmental isolates were performed and revealed a wide diversity of sequence types (STs). In Jordan, phenotypic and genetic features of E. coli were extensively studied but there is still a need to identify the STs that inhabit the community. METHODOLOGY: In this study, multi-locus sequence typing (MLST) was performed on archived clinical E. coli isolates collected from different hospitals in Jordan and the identified STs were extensively analyzed. RESULTS: Genotyping of 92 E. coli isolates revealed 34 STs and 9 clonal complexes. The frequencies of STs ranged between 1 to 23 observations. The most frequent STs among E. coli isolates were ST131 (n = 23), ST69 (n = 19), ST998 (n = 7), ST2083 (n = 5), and ST540 (n = 4). These five ST accounted for up to 60% of the 92 E. coli isolates. Based on the MLST database, the STs reported in this work were world widely recognized in humans, animals, and in the environment. CONCLUSIONS: This study has elaborated more knowledge about the genotypes of E. coli in Jordan, with recommendations for future studies to correlate its genotypes with virulence and resistance genes.

Anti_spike and anti_nucleocapsid IgG responses to SARS-CoV-2 in children of Jordan.

Background: It is proven that children have significantly milder COVID-19 disease compared to adults. Various immunological characteristics influence this age-related difference in protection against COVID-19. Pediatric COVID-19 in Jordan is extremely under reported. Objectives: The primary goal of this work is to identify the anti_S and anti_N antibody responses in a random group of children in Jordan and compare it to that of naturally infected-unvaccinated adults. Methods: 151 unvaccinated children, 4 days to 18 years old, were screened for anti_S and anti_N antibodies. History of COVID-19 infection or exposure to infection and symptom severity were reported by parents on a special questionnaire. Results: 78.9 % and 65.3 % of participants were seropositive for anti_S IgG and anti_N Abs, respectively. There was a remarkable association between age and anti_S IgG and anti_N IgG antibody titers, as children aged 12 years or older had increased anti_S IgG titers (mean = 19.3 BAU/mL) compared to younger groups (means of 10.15, 9.24, 7.91 BAU/mL for age groups 6-12, 1-6, less than 1 year, respectively). Gender did not show a statistically important role in anti_S and anti_N IgG seropositivity rates or titers. Children displayed significantly elevated anti_S titers (mean = 13.23 BAU/mL) compared to naturally infected adults (mean = 9.72 BAU/mL), in contrast, adults' anti_N titers (mean = 39.64 U/mL) were significantly higher compared to those of children (mean = 10.77 U/mL). Conclusions: The current work provides evidence of distinctly robust and persistent humoral immunity displayed by high anti_S and anti_N IgG in children, even >12 months post-infection. Age was the only factor that had a significant statistical impact on anti_S and anti_N Ab levels among the pediatric group in this study. Children exhibited significantly higher anti_S titers than naturally infected adults. In contrast, adults' anti_N titers were significantly higher. Such information can assist direct pediatric SARS-CoV-2 immunization programs, with implications for creating age-targeted strategies for diagnostic and population protection measures.

Coagulation-induced shedding of platelet glycoprotein VI mediated by factor Xa.

This study evaluated shedding of the platelet collagen receptor, glycoprotein VI (GPVI) in human plasma. Collagen or other ligands induce metalloproteinase-mediated GPVI ectodomain shedding, generating approximately 55-kDa soluble GPVI (sGPVI) and approximately 10-kDa platelet-associated fragments. In the absence of GPVI ligands, coagulation of platelet-rich plasma from healthy persons induced GPVI shedding, independent of added tissue factor, but inhibitable by metalloproteinase inhibitor, GM6001. Factor Xa (FXa) common to intrinsic and tissue factor-mediated coagulation pathways was critical for sGPVI release because (1) shedding was strongly blocked by the FXa-selective inhibitor rivaroxaban but not FIIa (thrombin) inhibitors dabigatran or hirudin; (2) Russell viper venom that directly activates FX generated sGPVI, with complete inhibition by enoxaparin (inhibits FXa and FIIa) but not hirudin; (3) impaired GPVI shedding during coagulation of washed platelets resuspended in FX-depleted plasma was restored by adding purified FX; and (4) purified FXa induced GM6001-inhibitable GPVI shedding from washed platelets. In 29 patients with disseminated intravascular coagulation, mean plasma sGPVI was 53.9 ng/mL (95% confidence interval, 39.9-72.8 ng/mL) compared with 12.5 ng/mL (95% confidence interval, 9.0-17.3 ng/mL) in thrombocytopenic controls (n = 36, P < .0001), and 14.6 ng/mL (95% confidence interval, 7.9-27.1 ng/mL) in healthy subjects (n = 25, P = .002). In conclusion, coagulation-induced GPVI shedding via FXa down-regulates GPVI under procoagulant conditions. FXa inhibitors have an unexpected role in preventing GPVI down-regulation.

Development of Novel Paper-Based Assay for Direct Serum Separation.

Background: Many conventional laboratory tests require serum separation using a clot activator/gel tube, followed by centrifugation in an equipped laboratory. The aim of this study is development of novel, equipment-free, paper-based assay for direct and efficient serum separation. Methods: Fresh blood was directly applied to wax-channeled filter paper treated with clotting activator/s and then observed for serum separation. The purity, efficiency, recovery, reproducibility, and applicability of the assay were validated after optimization. Results: Serum was successfully separated using activated partial thromboplastin time (APTT) reagent and calcium chloride-treated wax-channeled filter paper within 2 min. The assay was optimized using different coagulation activators, paper types, blood collection methods, and incubation conditions. Confirmation of serum separation from cellular components was achieved by direct visualization of the yellow serum band, microscopic imaging of the pure serum band, and absence of blood cells in recovered serum samples. Successful clotting was evaluated by the absence of clotting of recovered serum by prolonged prothrombin time and APTT, absence of fibrin degradation products, and absence of Staphylococcus aureus-induced coagulation. Absence of hemolysis was confirmed by undetectable hemoglobin from recovered serum bands. The applicability of serum separated in paper was tested directly by positive color change on paper using bicinchoninic acid protein reagent, on recovered serum samples treated with Biuret and Bradford reagents in tubes, or measurement of thyroid-stimulating hormone and urea compared to standard serum samples. Serum was separated using the paper-based assay from 40 voluntary donors and from the same donor for 15 days to confirm reproducibility. Dryness of coagulants in paper prevents serum separation that can be re-stored by a re-wetting step. Conclusions: Paper-based serum separation allows for development of sample-to-answer paper-based point-of-care tests or simple and direct blood sampling for routine diagnostic tests.

COVID-19-associated mental health impact on menstruation physiology: A survey study among medical students in Jordan.

BACKGROUND: The coronavirus disease 2019 pandemic has been an extraordinarily stressful situation in recent years. Stress is a physiological reaction to negative stimuli that is regulated by different neuroendocrine pathways. The female reproductive function is maintained by the menstrual cycle, which is negatively affected by hyperstimulation of stress signals. OBJECTIVES: This study evaluates the effect of the coronavirus disease 2019 outbreak on menstrual function and mental health, exploring the relationship between them. DESIGN: The current study uses a cross-sectional, survey-based design. METHODS: During this cross-sectional study, an online self-completion questionnaire was conducted among a sample of 385 Jordanian female medical students during the pandemic. The survey compared menstrual characteristics, depression, anxiety, and stress 10 months after the coronavirus disease 2019 pandemic with 10 months prior. Paired t-test, McNemar's test, Pearson's correlation, and multiple linear regression model were employed to analyze data using SPSS software. RESULTS: The mean age of female medical student respondents was 19.89 years. Data showed that the menstrual cycle length significantly increased during the coronavirus disease 2019 pandemic compared with 10 months prior (32.23 days versus 30.02 days, p = 0.019). The average number of heavy bleeding days also increased during the coronavirus disease 2019 pandemic (2.82 days versus 2.42 days, p = 0.002). The proportion of females with heavy bleeding amount was more than doubled during the pandemic of coronavirus disease 2019 compared with before (27.3% versus 10.4%, p = 0.000). Unpleasant menstrual signs such as nausea and/or vomiting, breast pain, and urinary urgency were significantly increased during the pandemic (p = 0.000, p = 0.008, and p = 0.024, respectively). During coronavirus disease 2019, a positive association between total Depression, Anxiety, and Stress Scale-21 Questionnaire score and heavy bleeding was identified (p < 0.05). The findings also indicated that mental disorders and the incidence of amenorrhea, nausea and/or vomiting, and urinary urgency were positively correlated during the coronavirus disease 2019 pandemic. The multiple regression analysis revealed associations between several menstrual characteristics such as amenorrhea and severity of bleeding with coronavirus disease 2019-related depression, anxiety, and stress. CONCLUSION: This study revealed that the stress related to the pandemic of coronavirus disease 2019 could affect the female menstrual cycle and hence the quality of women's life. Therefore, this study could serve as a baseline for planning and introducing stress mitigation interventions in crisis situations to improve the physiological and mental well-being of females and improve their quality of life.

Immunoglobulins response of COVID-19 patients, COVID-19 vaccine recipients, and random individuals.

BACKGROUND: The development of specific immunoglobulins to COVID-19 after natural infection or vaccination has been proposed. The efficacy and dynamics of this response are not clear yet. AIM: This study aims to analyze the immunoglobulins response among COVID-19 patients, COVID-19 vaccine recipients and random individuals. METHODS: A total of 665 participants including 233 COVID-19 patients, 288 COVID-19 vaccine recipients, and 144 random individuals were investigated for anti-COVID-19 immunoglobulins (IgA, IgG, IgM). RESULTS: Among COVID-19 patients, 22.7% had detectable IgA antibodies with a mean of 27.3±57.1 ng/ml, 29.6% had IgM antibodies with a mean of 188.4±666.0 BAU/ml, while 59.2% had IgG antibodies with a mean of 101.7±139.7 BAU/ml. Pfizer-BioNTech vaccine recipients had positive IgG in 99.3% with a mean of 515.5±1143.5 BAU/ml while 85.7% of Sinopharm vaccine recipients had positive IgG with a mean of 170.0±230.0 BAU/ml. Regarding random individuals, 54.9% had positive IgG with a mean of 164.3±214 BAU/ml. The peak IgM response in COVID-19 patients was detected early at 15-22 days, followed by IgG peak at 16-30 days, and IgA peak at 0-60 days. IgM antibodies disappeared at 61-90 days, while IgG and IgA antibodies decreased slowly after the peak and remained detectable up to 300 days. The frequency of IgG positivity among patients was significantly affected by increased age, admission department (inpatient or outpatient), symptoms, need for oxygen therapy, and increased duration between positive COVID-19 RT PCR test and serum sampling (p˂0.05). Positive correlations were noted between different types of immunoglobulins (IgG, IgM, and IgA) among patients. CONCLUSIONS: Natural infection and COIVD-19 vaccines provide IgG-mediated immunity. The class, positivity, mean, efficacy, and duration of immunoglobulins response are affected by the mechanism of immunity and host related variables. Random community individuals had detectable COVID-19 IgG at ~55%, far from reaching herd immunity levels.

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.

Chronotherapy in dentistry: A scoping review.

The circadian clock modulates almost all vital aspects of our physiology and metabolism, including processes relevant to dentistry, such as healing, inflammation and nociception. Chronotherapy is an emerging field aiming to improve therapeutic efficacy and decrease adverse effects on health outcomes. This scoping review aimed to systematically map the evidence underpinning chronotherapy in dentistry and to identify gaps in knowledge. We conducted a systematic scoping search using four databases (Medline, Scopus, CINAHL and Embase). We identified 3908 target articles screened by two blinded reviewers, and only original animal and human studies investigating the chronotherapeutic use of drugs or interventions in dentistry were included. Of the 24 studies included, 19 were human studies and five were animal studies. Chrono-radiotherapy and chrono-chemotherapy reduced treatment side effects and improved therapeutic response, leading to higher survival rates in cancer patients. Animal studies reported that tooth movement and periodontal tissue response to orthodontic forces follow a diurnal rhythm that might influence bone metabolism. Profound and prolonged local anesthesia could be achieved when injected in the evening. Although the overall quality of the included studies was low, chronotherapy applications in dentistry seem to have favourable outcomes, especially in head and neck cancer treatments.

Anti-S and Anti-N Antibody Responses of COVID-19 Vaccine Recipients.

The long-term immunoglobulin responses of COVID-19 vaccinations is important to determine the efficacy of these vaccinations. This study aimed to investigate and compare the long-term immunoglobulin response of COVID-19 vaccination recipients, using anti-S IgG, anti-N IgG, and IgM titer levels. This study included 267 participants, comprising individuals who tested positive for COVID-19 through PCR testing (n = 125), and those who received the Pfizer (n = 133), Sinopharm (n = 112), AstraZeneca (n = 20), or Sputnik (n = 2) vaccines. Female participants comprised the largest share of this study (n = 147, 55.1%). This study found that most participants had positive IgG antibodies, with 96.3% having anti-S IgG and 75.7% having anti-N IgG. Most participants (90.3%) tested negative for anti-N IgM antibodies. Sinopharm-vaccinated individuals exhibited a notably lower rate of positive anti-S IgG (93.8%) and a significantly higher rate of positive anti-N IgG antibodies (91%). Anti-N IgG levels were significantly correlated with the number of prior COVID-19 infections (p = 0.015). Specifically, individuals with a history of four COVID-19 infections had higher anti-N IgG titers (14.1 ± 1.4) than those with only one experience of COVID-19 infection (9.4 ± 7.2). Individuals who were infected with COVID-19 after receiving the vaccine demonstrated higher levels of anti-N IgG, exhibiting a 25% increase in mean titer levels compared to those who were infected prior to vaccination. There was a statistically significant association between anti-N IgG positivity with age (p = 0.034), and smoking status (p = 0.006) of participants. Participants younger than 20 and older than 60 showed the highest positivity rate of anti-N (>90%). Smokers had a low positivity rate of anti-N (68.8%) compared to nonsmokers (83.6%). In conclusion, this study demonstrated that most COVID-19 vaccination recipients had positive IgG antibodies, with differences in the long-term immunoglobulin response depending on the type of vaccine administered and occurrence of COVID-19 infection.

Validation of paper-based assay for rapid blood typing.

We developed and validated a new paper-based assay for the detection of human blood type. Our method involves spotting a 3 µL blood sample on a paper surface where grouping antibodies have already been introduced. A thin film chromatograph tank was used to chromatographically elute the blood spot with 0.9% NaCl buffer for 10 min by capillary absorption. Agglutinated red blood cells (RBCs) were fixed on the paper substrate, resulting in a high optical density of the spot, with no visual trace in the buffer wicking path. Conversely, nonagglutinated RBCs could easily be eluted by the buffer and had low optical density of the spot and clearly visible trace of RBCs in the buffer wicking path. Different paper substrates had comparable ability to fix agglutinated blood, while a more porous substrate like Kleenex paper had enhanced ability to elute nonagglutinated blood. Using optimized conditions, a rapid assay for detection of blood groups was developed by spotting blood to antibodies absorbed to paper and eluted with 200 µL of 0.9% NaCl buffer directly by pipetting. RBCs fixation on paper accurately detected blood groups (ABO and RhD) using ascending buffer for 10 min or using a rapid elution step in 100/100 blood samples including 4 weak AB and 4 weak RhD samples. The assay has excellent reproducibility where the same blood group was obtained in 26 samples assessed in 2 different days. Agglutinated blood fixation on porous paper substrate provides a new, simple, and sensitive assay for rapid detection of blood group for point-of-care applications.

COVID-19-Associated Mental Health Impact on Menstrual Function Aspects: Dysmenorrhea and Premenstrual Syndrome, and Genitourinary Tract Health: A Cross Sectional Study among Jordanian Medical Students.

The physiology of reproduction is affected by psychological distress through neuroendocrine pathways. Historically, COVID-19 is one of the most stressful events with devastating consequences. This research aims to investigate the relationship between dysmenorrhea, PMS, and reproductive tract health on one hand, and COVID-19-related anxiety, depression, and stress on the other among medical students in Jordan. Medical students were invited through teaching platforms and social media to complete an online survey. SPSS software was used to analyze data. A total of 385 medical students participated in this research. Hence, 49.9% of the study population reported severe dysmenorrhea during COVID-19 compared to 36.9% before COVID-19 (p = 0.000). Dysmenorrhea was significantly associated with disruptions of sport and daily activities during COVID-19 (p = 0.015 and p = 0.002, respectively). The prevalence of PMS components, e.g., mastalgia, fatigue, headache, palpitation, and emotional and sleep disturbances, was raised during COVID-19 compared with before (p < 0.05). Symptoms of genitourinary tract infections, such as lower abdominal pain, vaginal discharge, genitalia rash/ulcers and itching, and urgency, were significantly increased after COVID-19 (p < 0.05). Positive Pearson correlations between COVID-19-associated mental health disorders and dysmenorrhea severity, PMS, and genital tract health abnormalities were observed (p < 0.05). The multiple linear regression model revealed that dysmenorrhea severity, PMS symptoms like palpitation, and genitourinary symptoms like lower abdominal pain and urgency were associated with worsening of depression, while dysuria was associated with a protective effect against depression. Moreover, it was observed that dysmenorrhea severity, PMS symptoms, such as headache and palpitation, and urinary urgency were associated with aggravation of anxiety. However, food craving and dysuria were protective against anxiety. Finally, dysmenorrhea severity, PMS symptoms of headache and palpitation, lower abdominal pain, and urgency were related to worsening of stress, whereas the premenstrual symptom of breast pain was a protective factor against stress. This work showed that COVID-19 pandemic-related psychological distress and menstrual, premenstrual, and genitourinary symptoms are closely related. Further future work is required to evaluate the long lasting-effects of the pandemic on mental health and the physiology of reproduction.

Efficient and Simple Paper-Based Assay for Plasma Separation Using Universal Anti-H Agglutinating Antibody.

BACKGROUND: Conventional laboratory tests require plasma separation using centrifugation by skilled personnel in well-equipped lab. Development of a simple, reliable, and cheap point-of-care (POC) test for plasma separation will overcome these limitations. METHODS: Plasma separation was achieved in filter paper using the anti-H agglutinating antibody. Hydrophobic channels were created using a solid ink printer. The reproducibility, efficiency, recovery, and applicability of the assay were validated on a large number of blood samples. RESULTS: A simple, fast, cheap, and direct paper-based assay for plasma separation from whole blood using universal anti-H agglutinating antibody was developed without equipment or pretreatment requirements. The purity of plasma separation using anti-H treated paper was confirmed by microscopy and biuret test for plasma albumin detection. Plasma separation was affected by paper structure, antibody concentration, donor gender, and hematocrit. The efficiency of the assay was 72% and the reproducibility was about 90% with minimal interassay and intra-assay variabilities. The assay successfully separated plasma from 116/119 samples, indicating high sensitivity (97.5%). Furthermore, the assay accurately recovers thyroid stimulating hormone from samples compared to standard methods with 107% recovery rate. CONCLUSIONS: Paper-based plasma separation using anti-H agglutinating antibodies would have numerous applications in paper-based POC tests and in resource limited areas.

SARS-CoV-2 Antinucleocapsid Antibody Response of mRNA and Inactivated Virus Vaccines Compared to Unvaccinated Individuals.

Comparative studies of SARS-CoV-2 antinucleocapsid (anti-N) antibody response in the context of inactivated virus vaccines versus natural infection are limited. This study aims to determine and compare the anti-N antibody levels in people vaccinated with Sinopharm's (Wuhan, China) inactivated virus vaccine in comparison with naturally infected unvaccinated and Pfizer's spike (S) mRNA-based vaccinated subjects. Two hundred ninety-nine Jordanian adults participated in the study including unvaccinated COVID-19-infected patients (n = 99), Pfizer-vaccinated (n = 100), and Sinopharm-vaccinated recipients (n = 100). Serum samples were assayed for anti-N IgG, anti-N IgM, and anti-S IgG. Sera of 64.6% of naturally infected unvaccinated participants had positive anti-S IgG (median = 36.35 U/mL; range: 0.04−532.5 U/mL) compared to 88% of Pfizer-vaccinated (Manhattan, NY, USA) (median = 26.52 U/mL; range: 0.39−1265 U/mL) and 58% of Sinopharm-vaccinated subjects (median = 14.35 U/mL; range: 0.39−870.17 U/mL). Samples of 60.6% of naturally infected unvaccinated people had positive anti-N IgG (median = 15.03 U/mL; range: 0−265.1 U/mL) compared to 25% of Pfizer-vaccinated (median = 0.02 U/mL; range: 0−68 U/mL) and 48% of Sinopharm-vaccinated subjects (median = 0.8 U/mL; range: 0−146.3 U/mL). Anti-N titers among the three groups were significantly different (p < 0.05). Anti-N IgM antibodies appeared in 23.2% of the naturally infected unvaccinated group (median = 0.29 U/mL; range: 0−15 U/mL) compared to only 9.0% of Pfizer-vaccinated (median = 018 U/mL; range: 0−33 U/mL) and 7.0% of Sinopharm-vaccinated subjects (median = 0.2 U/mL; range: 0−12.02 U/mL). A significant negative correlation was found between anti-S and age for both vaccines and between anti-S and the presence of chronic disease in Sinopharm-vaccinated subjects. A significant positive correlation between anti-N and anti-S titers was found among the three groups. This study shows that the inactivated virus vaccine, Sinopharm, induces an anti-N response that can boost that of natural infection or vice versa. On the other hand, the Pfizer mRNA-based vaccine induces a significantly stronger anti-S Ab response.

Multidrug-Resistant Acinetobacter baumannii in Jordan.

Background: Acinetobacter baumannii is a common cause of multi-drug (MDR)-resistant infections worldwide. The epidemiological and molecular characteristics of MDR-A. baumannii in Jordan is not known. Methods: A. baumannii isolates were collected from 2010 to 2020 from three tertiary hospitals in Jordan. Demographic and clinical data, isolates information, antibiotic susceptibility patterns, phenotypic, and molecular characterization of carbapenem resistance genes were performed. Results: A total of 622 A. baumannii isolates were collected during the study period. Most isolates were from males, aged 18−60 years, Jordanian, from infected wounds, and were patients in surgery or critical care units. Among patients from whom A. baumannii was isolated, associated risk factors for MDR were adults over 60, males, critically ill patients and infected wounds (OR 4.14, 2.45, 10, 7, respectively, p < 0.0001). Incidence rates from 2010 to 2015 showed a slight increase in MDR (3.75/1000 to 4.46/1000). Resistance patterns indicated high resistance for most cephalosporins, carbapenems, and fluoroquinolones, moderate resistance for trimethoprim/sulfamethoxazole and ampicillin/sulbactam, low resistance for aminoglycosides and tetracyclines, while colistin and tigecycline, have the lowest resistance rates. 76.8% of A. baumannii isolates were MDR and 99.2% were carbapenem-resistant. All isolates were positive for the OXA-51 gene (100%), 98.5% were positive for the OXA-23 gene, 26.6% for the VIM gene, while KPC and IMP genes were almost not detected (0% and 0.8% respectively). Conclusions: This is the first large, multicentric, prolonged study that provides insights into A. baumannii infections in Jordan. Attention to patients at higher risk is important for early identification. Colistin and tigecycline were the most effective antimicrobials.