The severity of COVID-19's outcomes has been positively correlated with an increased risk of respiratory failure and death, especially in patients with chronic illnesses. This case-control design study aims to examine the correlation in the Palestinian population in light of its impact on diabetic patients. The study was conducted from March 2020 to June 2021 on 417 patients admitted to the Palestinian National Center for Rehabilitation. Of them, 198 cases were tested positive for COVID-19 and had diabetes, whereas the remaining 219 were those who tested positive for COVID-19 but were not diagnosed with diabetes and acted as controls. Data from patient files were collected to address the study questions. Patients' ages ranged from 17 to 98 years, with a mean age of 58. Male participants represented 53.5% of the total. The results of the current study indicated that the case fatality rate (CFR) was 14.2% for all participants and 19.7% for patients with diabetes. In regard to patients' health conditions, 5.7% had cardiovascular diseases (CVD), 33.6% had hypertension, 6.5% had kidney diseases, and 47.4% had diabetes. According to the multivariate analysis, diabetic patients had a 1.63 times higher risk for COVID-19 infection compared to nondiabetic patients (adjustment of odds ratio of 1.63 and confidence interval [CI] = 0.354-1.856). This is the first study to investigate the relationship between the severity of COVID-19 outcomes in Palestine and the diabetes status, and the majority of its findings are consistent with other research studies that assessed the impact of COVID-19 on diabetic patients (with minor variations in the number and percentage). The study confirms that along with diabetes, age, hypertension, kidney diseases, shortness of breath, requirement of oxygen, D-dimer, C-reactive protein (CRP), kidney function test (KFT), and days of hospitalization were also associated with severe COVID-19 outcomes, whereas gender, CVDs, and liver function test (LFT) were not associated with severe COVID-19 outcomes.
Background. Salmonella enterica serovar Typhimurium (Salmonella Typhimurium) is a common food-borne pathogen that causes gastroenteritis and can lead to life-threatening systemic disease when it spreads to vital organs, such as the liver. Stimulator of interferon genes (STING) is a crucial regulator of the host’s innate immune response to viral infections, while its role in bacterial infections remains controversial. This study aims to establish a STING-deficient HepG2 cell line through the CRISPR/Cas9 system and evaluate its effects on Salmonella replication. Methods. In this study, a STING knockout HepG2 cell line was constructed through the application of CRISPR/Cas9 technology. We assessed cell viability and proliferation using the CCK-8 assay. Subsequently, we investigated the effect of STING deletion on Salmonella replication and the expression of type I interferon-related genes. Results. The STING knockout HepG2 cell line was successfully constructed using the CRISPR/Cas9 system. The proliferation capability was diminished in STING-deficient HepG2 cells, while Salmonella Typhimurium replication in these cells was augmented compared to the wild-type (WT) group. Following Salmonella infection, the transcriptional responses of type I interferon-related genes, such as IFNB1 and ISG15, were inhibited in STING-deficient HepG2 cells. Conclusions. We successfully constructed a STING-deficient cell line. Our finding of increased Salmonella Typhimurium replication in STING-deficient HepG2 cells provides the basis for further studies on pathogen-host interactions.
Aim. The increasing antibiotic resistance and the ability to form biofilms in medical devices have become the leading cause of severe infections associated with Staphylococcus aureus (S. aureus). Since the bacteria living in biofilms can exhibit 10- to 1,000-fold increase in antibiotic resistance and implicate chronic infectious diseases, the detection of S. aureus ability to form biofilms is of great importance for managing, minimizing, and effectively treating infections caused by it. This study aimed to compare the tube and tissue culture methods to detect biofilm production and antibiotic susceptibility in MRSA and MSSA. Materials and Methods. The S. aureus isolates were identified by the examination of the colony morphology, Gram staining, and various biochemical tests. Antimicrobial susceptibility testing of all isolates was performed by the modified Kirby–Bauer disc diffusion method as recommended by CLSI guidelines. MRSA screening was performed phenotypically using a cefoxitin disc (30 µg). Isolates were tested for inducible resistance using the D-test, and two phenotypic methods detected biofilm formation. Results. Among 982 nonrepeated clinical specimens, S. aureus was isolated from 103 (10.48%). Among 103 clinical isolates of S. aureus, 54 (52.42%) isolates were MRSA, and 49 (47.57%) were MSSA. Among 54 MRSA isolates, the inducible MLSB phenotype was observed in 23/54 (42.59%) with a positive D-test. By TCP method, 26 (48.1%) MRSA isolates were strong biofilm producers, whereas, among all MSSA isolates, only 6 (12.2%) were strong biofilm producers. Conclusion. MRSA showed strong biofilm production in comparison with MSSA. The TCP method is a recommended reliable method to detect the biofilm among S. aureus isolates, and the TM method could be useful for the screening of biofilm production in S. aureus in the routine clinical laboratory.
Pseudomonas aeruginosa, a Gram-negative opportunistic bacterium, has emerged as a cause of life-threatening infections in burn wounds. Current therapeutic approaches through wound dressings and systemic medicines are far from satisfactory; resistance to more than two antibiotics shown by pathogens contributes to failures of therapy causing mortality. This animal study was conducted to check the efficacy of one Ayurveda-based fermented polyherbal preparation (AP 01) against multiple antibiotics-resistant (MAR) P. aeruginosa HW01-infected rat burn wounds. AP-01 was applied on artificially infected burn wound on a rat model infected with MAR P. aeruginosa to register the healing effects in terms of reduction in residual wound area percentage, the presence of C-reactive protein in blood, and the presence of viable bacteria colony. Topical application with conventional antibiotics served as a positive control. The polyherbal preparation had reduced the infected residual burn wound area at 40.63% ± 0.69 from the initial burn wound area within two weeks after a single intervention, whereas residual burn wound area remained much higher in the case of animals left untreated and in the case of the animals treated with control drug. Restoration to the normalcy of serum C-reactive protein level was also achieved earlier in the case of polyherbal AP-01-treated groups than in other groups. Fermented formulations using components of AP-01 singly or in different combinations had never been tested earlier for topical application in infected burn wound. The formulation of AP-01 was found superior in terms of the rate of healing and control of infection by MAR P. aeruginosa strains in burn wounds in rat models.
The involvement of multiple antibiotic-resistant gastrointestinal pathogens in diarrhea aggravates the disease condition uncontrollably. The current study aimed to find and develop a suitable formulation utilizing multiple natural components from known plant sources to augment the current therapeutic outcomes. The hydroethanolic extraction method was applied through boiling and fermentation on ancient observation-based efficacious plant parts for developing the antidiarrheal polyherbal formulation AP-01. An animal study model of diarrhea was used to evaluate the safety and efficacy of the formulation. The formulation was tested in vitro on four different multiple antibiotic-resistant gastrointestinal pathogens collected from the national repository. The formulation depicted no cytotoxicity on normal gut cells and was efficacious at 10 ml/kg single dose in relieving symptoms of diarrhea by 79.71%, compared with the standard drug showing a reduction of symptoms by 83.01%. AP-01 exhibited delayed gastric motility. The symptoms of diarrhea ceased to occur within 320.66 ± 5.05 minutes with AP-01, whereas the standard drug took 308 ± 6.63 minutes. AP-01 was found successful at a viable dosage regimen of 75 μl/ml v/v to 100 μl/ml v/v in inhibiting the growth of different pathogens from the Enterobacteriaceae family possessing resistance against several classes of antibiotics in culture media. Chemical analysis revealed different alkaloids, flavonoids, and polyphenols that probably work in unison through multiple modes of action to arrest diarrhea and inhibit pathogens simultaneously. These promising results shown by AP-01 should evoke an effort to dive deep into research and development for better therapeutic formulations for infectious diarrhea by harvesting nature's arsenal.
