Simultaneous Detection of Influenza A/B, Respiratory Syncytial Virus, and SARS-CoV-2 in Nasopharyngeal Swabs by One-Tube Multiplex Reverse Transcription Polymerase Chain Reaction.

The treatment and outcome of respiratory virus infections differ. SARS-CoV-2, as well as other respiratory viruses such as influenza virus (A and B) and respiratory syncytial virus (RSV), require simultaneous, cost-effective, and rapid differential detection. We used a gold standard five-target single-step RT-PCR to detect influenza viruses, RSV, and SARS-CoV-2, and this method can be extended to detect influenza virus subtypes. As a result, this five-target single-step RT-PCR method is ideal for differentiating respiratory viruses. The 5' nuclease activity of Taq DNA polymerase is used in the real-time reverse transcription PCR assay. The Taq man fast viral 1-step enzyme is a 4× Master mix and five-target primer probe mix that detects influenza A, influenza B, SARS-CoV-2 ORF1ab, respiratory syncytial viruses A/B and actin. When compared with TaqMan TM and Invitrogen superscript TM III Platinum and the Meril Kit for SARS-CoV-2, the assay demonstrated 100% sensitivity, specificity, and amplification efficiency of 90.1% for target genes. In conclusion, our one-tube multiplex RT-PCR assay offers a rapid and reliable method for the simultaneous detection of influenza A/B, RSV, and SARS-CoV-2 from nasopharyngeal swabs. This assay has the potential to enhance diagnostic capabilities and improve public health responses during respiratory outbreaks, enabling timely interventions and informed decision making.

Resistance Pattern in Mostly Gram-negative Bacteria Causing Urinary Tract Infections.

PURPOSE: The antimicrobial prescription in urinary tract infections (UTI) is driven by local data on its pathogenic spectrum and the resistance pattern exhibited by the disease-causing pathogens. We aimed to determine the bacteriological diversity of UTI causing pathogens and antimicrobial resistance in mostly gram-negative bacteria. METHODS: This retrospective hospital-based cross-sectional study analyzed the culture and sensitivity reports of urine samples from a referral centre in the Aljouf region of Saudi Arabia. All the antibiograms from January 1, 2020, to December 31st 2020, were included. The bacterial identification and antimicrobial testing were carried out by the BD Phoenix system (BD Diagnostics, Sparks, MD, USA). Antimicrobial testing was performed as per the Clinical and Laboratory Standard Institute recommendations. Frequencies of multidrug- and extensive drug resistance were calculated. RESULTS: Of the 1334 non-duplicate urine samples received, 422 (31.6%) bacterial growths were observed. Of these, 383 (90.8%) and 39 (9.2%) were gram-negative and gram-positive bacterial isolations, respectively. E. coli 161 (38.1%), K. pneumoniae 97 (23.0%), and E. faecalis 18 (4.3%) were frequent aetiologies of UTI. 309 (80.7%) of gram-negative bacteria were multidrug-resistant including 88 (23.0%) extensively drug-resistant. Overall, a resistance rate of > 55 % to 1st through 4th generation cephalosporins was observed except for cefoxitin (43.7%). A resistance rate of 37.6% was observed towards carbapenems, with the lowest rate (34.0%) to meropenem. CONCLUSION: Multi-drug resistant gram-negative bacteria dominate the pathogenic spectrum of UTI in the region. A high resistance rate to cephalosporins and carbapenems exists in gram-negative organisms, causing UTI.

Clinical characteristics among patients with COVID­19: A single­center retrospective study.

The aim of the present study was to investigate the clinical features and laboratory parameters of hospitalized patients with coronavirus disease 2019 (COVID-19) and assess the characteristics between severe and non-severe cases. The study retrospectively analyzed the clinical data of 1,096 patients, of which, 626 (57.11%) and 470 (42.89%) were categorized into severe and non-severe groups, respectively. Clinical parameters such as signs and symptoms, comorbidities, levels of D-dimer, C-reactive protein (CRP), interleukin 6 (IL-6) and lactate dehydrogenase were analyzed. The data are presented as frequencies, means and standard deviations. The chi-square test and Mann-Whitney U test were used to assess any significant differences between the severe and non-severe COVID-19 groups. The clinical symptoms in severe COVID-19 cases included anosmia (P≤0.01), sore throat (P≤0.01), fatigue (P≤0.01), headache (P≤0.01), and shortness of breath (P≤0.01). Laboratory findings showed a significant increase in CRP (21.90±40.23 vs. 16.13±21.82; P≤0.01) and IL-6 levels (58.92±55.07 vs. 41.41±38.30; P≤0.01). Patients with severe COVID-19 had significant lymphopenia compared with that in non-severe cases. Among the comorbidities, hypertension (P≤0.01) was significantly more frequent in patients with severe COVID-19. In conclusion, major derangements in laboratory parameters were observed in patients with severe COVID-19 infection.

Quantitative Changes in White Blood Cells: Correlation with the Hallmarks of Polycystic Ovary Syndrome.

Background and Objectives: In women of reproductive age, leukocytosis is a risk factor that bridges low-grade chronic inflammation (metabolic inflammation), metabolic changes, and polycystic ovary syndrome (PCOS) and is a potential early predictor of PCOS. This study aims to explore the predictive role of quantitative changes in white blood cells (WBCs) and neutrophils in PCOS-associated metabolic changes. Materials and Methods: A total number of 176 blood samples were obtained from age-matched women of the reproductive period, comprising 88 PCOS cases and 88 healthy controls. Hematological, metabolic, and anthropometric indices and ultrasonic assessment were recorded. Results: Elevated levels of luteinizing hormone, testosterone, and lipid parameters except HDL-C levels, and the prevalence of metabolic syndrome in PCOS were statistically significant (p < 0.001). The neutrophil count and neutrophil−lymphocyte ratio (NLR) in PCOS patients were significantly higher (p < 0.001) than their counterparts. The predictive ability of the neutrophil count and neutrophil−lymphocyte ratio (NLR) for PCOS, and possibly its associating subclinical inflammation at optimum cut-off values for the neutrophil count and NLR of >46.62% (sensitivity 94.32% and specificity 74.42%) and >1.23 (sensitivity 71.59% and specificity 100%), respectively. With regard to the areas under the curve (AUC) and Youden indices, they constituted 0.922 and 0.697 for neutrophil count and 0.926 and 0.716 for NLR, respectively. The comparative ROC z-statistic value was 2.222 and a p = 0.026. The multiple linear regression analysis revealed no significant influence for hormonal and metabolic independent variables on the neutrophil count in PCOS cases, but, as can be expected, revealed a significant negative relationship with the other components of WBCs. Conclusion: In conclusion, relative neutrophilia and elevated NLR are potential cost-effective, sensitive, and specific predictors of PCOS that may also shed light on the mechanism of chronic low-grade inflammation that is characteristic of the disease.

Bacteriological profile of wound infections and antimicrobial resistance in selected gram-negative bacteria.

Background: Managing wound infections is a challenging task. Understanding their resistance pattern is an essential step at reducing its burden in hospital settings. Objective: To determine the bacteriological diversity of wound infections and the antimicrobial resistance exhibited by a selected Gram-negative bacterium in the Aljouf region of Saudi Arabia. Methods: The study retrospectively analysed the antibiograms of wound infections from hospitalized patients for the year 2019. The European Centre for Disease Control guidelines were adopted for the classification of resistant bacteria. Multidrug-, extensive drug-, and carbapenem-resistant isolates are presented as frequencies and percentages. Results: A total of 295 non-duplicate wound swab antibiograms were retrieved, 64.4% (190) and 35.6% (105) isolates were Gram-negative and Gram-positive bacterial infections respectively. Predominant pathogens included Staphylococcus species 21.0% (62), E. coli 16.3% (48) and K. pneumoniae 13.5% (40). 148 (77.9%), 42 (22.1%) and 43 (22.6%) of the Gram-negative isolates were multidrug-, extensively drug- and carbapenem-resistant. The antibiotic resistance exhibited by gram-negative bacteria was 43.4% (234/539), 59.1% (224/379) and 53.7% (101/188) towards carbapenems, 3rd - and 4th - generation cephalosporins. Conclusions: The majority of wound infections are caused by multidrug-, extensively drug- and carbapenem-resistant Gram-negative bacteria. Further studies should focus on the molecular basis of this resistance.

Resistance Patterns of Gram-Negative Bacteria Recovered from Clinical Specimens of Intensive Care Patients.

Intensive care units are complex environments favoring high resistance in microorganisms. This study evaluated the resistance and the distribution dynamics of resistant Gram-negative bacteria (GNB) in patients admitted to intensive care units. This retrospective, record-based, cross-sectional study analyzed all of the antibiograms of patients admitted to the ICUs. The BD Phoenix system (BD Diagnostics, Sparks, MD, USA) was used for bacterial identification and antimicrobial testing. Clinical and Laboratory Standard Institute recommendations were used for antimicrobial testing. Frequencies and percentages of multidrug and pan-drug resistance were calculated. A total of 570 bacterial growths were observed, out of which 437 (76.7%) were of GNB. K. pneumoniae (21.0%), P. aeruginosa (11.8%), and Staphylococcus aureus (13.2%) were the most frequent disease-causing bacteria in intensive care patients. Resistance rates of 73.2% and 70.1% were observed for third- and fourth-generation cephalosporins, respectively, while 48.2% carbapenem and > 65% fluoroquinolones resistance rates were observed. Amikacin was the most effective antibiotic, with a sensitivity rate of 69.5%. A total of 372 (85.1%) of GNB were multidrug resistant. The majority of infections in intensive care patients are caused by multidrug-resistant (MDR) Gram-negative bacteria. Female gender and advancing age are factors favoring MDR. Enhanced surveillance and strengthening of the antimicrobial stewardship program are warranted.

ESBL Activity, MDR, and Carbapenem Resistance among Predominant Enterobacterales Isolated in 2019.

Antimicrobial-resistance in Enterobacterales is a serious concern in Saudi Arabia. The present study retrospectively analyzed the antibiograms of Enterobacterales identified from 1 January 2019 to 31 December 2019 from a referral hospital in the Aljouf region of Saudi Arabia. The revised document of the Centers for Disease Control (CDC) CR-2015 and Magiorakos et al.'s document were used to define carbapenem resistance and classify resistant bacteria, respectively. The association of carbapenem resistance, MDR, and ESBL with various sociodemographic characteristics was assessed by the chi-square test and odds ratios. In total, 617 Enterobacterales were identified. The predominant (n = 533 (86.4%)) isolates consisted of 232 (37.6%), 200 (32.4%), and 101 (16.4%) Escherichia coli, Klebsiella pneumoniae, and Proteus mirabilis, respectively. In general, 432 (81.0%) and 128 (24.0%) isolates were of MDR and ESBL, respectively. The MDR strains were recovered in higher frequency from intensive care units (OR = 3.24 (1.78-5.91); p < 0.01). E. coli and K. pneumoniae resistance rates to imipenem (2.55 (1.21-5.37); p < 0.01) and meropenem (2.18 (1.01-4.67); p < 0.04), respectively, were significantly higher in winter. The data emphasize that MDR isolates among Enterobacterales are highly prevalent. The studied Enterobacterales exhibited seasonal variation in antimicrobial resistance rates towards carbapenems and ESBL activity.

Prevalence and Molecular Analysis of Novel Newly Identified Periodontal Pathogens in Subgingival Plaque Samples of Saudi Patients with Chronic Periodontitis

Objective: To evaluate the prevalence of novel newly identified periodontal pathogenic strains in subgingival plaque samples and relate it with bleeding on probing, probing pocket depth and age. Material and Methods: 268 chronic periodontitis patients with a mean age of 46.0 ± 6.0 years were included. The following microorganisms were evaluated: Campylobacter gracilis (Cg), Fusobacterium nucleatum (Fn), Porphyromonas gingivalis (Pg), and Tannerella forsythia (Tf). Full mouth examination was registered; the probing pocket depth and clinical attachment level were assessed at six sites per tooth. Dental subgingival plaque samples were taken in the deepest pocket per arch in the maxilla and mandible. DNA analysis was performed using DNA-strip technology. Pearson's correlation coefficient was used for statistical analysis. Results: Porphyromonas gingivalis and Tannerella forsythia were detected at high level of 80% and 82% respectively. F nucleatum revealed a rate of 94%. Bacterial load significantly increased by increasing mean probing depth of the examined sites. Pearson's coefficient was the highest for Pg (r=0.62, p=0.0001) and the lowest for Cg (r=0.08, p=0.04). The bacterial load significantly increased by increasing the number of bleeding sites; Pearson's coefficient varied from r= 0.14 for Pg (p=0.01) to r=0.39 for Tf (p=0.001). Conclusion: It was confirmed the presence of main putative periodontal pathogens detectable in Saudi periodontal subjects, also providing a comprehensive view for correlation of these putative periodontal pathogens with the increase in probing pocket depth to the presence and load of all the bacterial species.

Antiurolithic effect of olive oil in a mouse model of ethylene glycol-induced urolithiasis.

PURPOSE: At present, commercially available antiurolithic drugs have more adverse effects than potential therapeutic or preventive effects with chronic use. With this in mind, the present study was designed to assess the antiurolithic effect of olive oil in a mouse model of ethylene glycol (EG)-induced urolithiasis. MATERIALS AND METHODS: Adult albino mice were divided into 6 groups. Group I was fed the vehicle only. Group II was supplemented with 0.75% EG alone in drinking water during the experimental period to initiate deposition of calcium oxalate in kidneys, which leads to urolithiasis in animals. Groups III (olive oil control group) through V were fed olive oil orally at various doses during the experimental period. Group VI received cystone (750 mg/kg). Groups IV-VI additionally received 0.75% EG in drinking water ad libitum. SPSS ver.17.0 was used for statistical analysis. RESULTS: The study results showed significantly higher levels of serum urea, uric acid, and creatinine (p<0.05) in group II than in groups III-VI and I. Administration of olive oil at different doses restored the elevated serum parameters in groups IV and V compared with group II. Urine and kidney calcium, oxalate, and phosphate levels in groups IV-VI were significantly lower (p<0.05) than in animals with EG-induced urolithiasis (group II). Group V mice showed a significant restoration effect on serum as well as urine and kidney parameters compared with group II. CONCLUSIONS: Supplementation with olive oil (1.7 mL/kg body weight) reduced and prevented the growth of urinary stones, possibly by inhibiting renal tubular membrane damage due to peroxidative stress induced by hyperoxaluria.

Variation of antioxidant activity and phenolic content of some common home remedies with storage time.

The variation of antioxidant activity and total phenolic content of some plants commonly used as home remedies were screened on the basis of air drying time of (short term and long term air) the dried plant Ocimum sanctum (leaf), Cucumis sativus (Seed), and ethanolic extracts of Capsicum frutescens (Seed) and Coriandrum sativum (Seed). Antioxidant activities and total phenolic content values were estimated using DPPH reagent and Folin Ciocalteu reagent methods respectively. The antioxidant activity in short term air dried Ocimum sanctum plant methanol extract (IC50 = 0.03 ± 0.01 mg/l) was the highest among the other plant extracts. However, Coriandrum sativum plant ethanol extract produced higher phenolic content (70.24±3.4/66.57±4.1 mg/g) than other plant extracts. It was found that there was no correlation between antioxidant and phenolic content of these plant extracts.

Comparative study of antimicrobial activity and phytochemical analysis of methanolic and aqueous extracts of the fruit of Emblica officinalis against pathogenic bacteria.

OBJECTIVE: To investigate antimicrobial potential of aqueous (infusions, decoctions) and methanlic extracts (1:2 and 1:5 concentrations) of Emblica officinalis (amla) against seven pathogenic bacteria namely Staphylococcs aureus, Staphylococcus saprophyticus, Escherichia coli, Enterococcus faecalis, Enterococcus cloacae, Proteus vulgaris and Klebsiella pneumoniae. METHODS: The well diffusion technique was employed. The minimum inhibitory concentration (MIC) was determined using micro-broth dilution methods and phytochemical screening was done as per standard procedures. RESULTS: Aqueous infusion extract of amla exhibited potent antimicrobial activity against E. cloacae followed by E. coli. Preliminary phytochemical analysis of E. officinalis aqueous extracts (infusions and decoctions) only showed presence of tannins, saponins, flavanoids, Terpenoids and phenols. MIC of aqueous extract of E. officinalis was most active against K. pneumoniae. Whereas MIC of methanol extract of E. officinalis shows maximum activity against E. coli. CONCLUSION: Emblica officinalis definitely possesses potent antimicrobial activities and this can serve as an important platform for the development of inexpensive, safe and effective medicines