Mutations of SARS-CoV-2 spike protein: Implications on immune evasion and vaccine-induced immunity.

Responsible for more than 4.9 million deaths so far, COVID-19, caused by SARS-CoV-2, is instigating devastating effects on the global health care system whose impacts could be longer for the years to come. Acquiring a comprehensive knowledge of host-virus interaction is critical for designing effective vaccines and/or drugs. Understanding the evolution of the virus and the impact of genetic variability on host immune evasion and vaccine efficacy is helpful to design novel strategies to minimize the effects of the emerging variants of concern (VOC). Most vaccines under development and/or in current use target the spike protein owning to its unique function of host receptor binding, relatively conserved nature, potent immunogenicity in inducing neutralizing antibodies, and being a good target of T cell responses. However, emerging SARS-CoV-2 strains are exhibiting variability on the spike protein which could affect the efficacy of vaccines and antibody-based therapies in addition to enhancing viral immune evasion mechanisms. Currently, the degree to which mutations on the spike protein affect immunity and vaccination, and the ability of the current vaccines to confer protection against the emerging variants attracts much attention. This review discusses the implications of SARS-CoV-2 spike protein mutations on immune evasion and vaccine-induced immunity and forward directions which could contribute to future studies focusing on designing effective vaccines and/or immunotherapies to consider viral evolution. Combining vaccines derived from different regions of the spike protein that boost both the humoral and cellular wings of adaptive immunity could be the best options to cope with the emerging VOC.

The role of long noncoding RNAs in the diagnosis, prognosis and therapeutic biomarkers of acute myeloid leukemia.

Acute myeloid leukemia (AML) is the abnormal proliferation of immature myeloid blast cells in the bone marrow. Currently, there are no universally recognized biomarkers for the early diagnosis, prognosis and effective treatment of AML to improve the overall survival of patients. Recent studies, however, have demonstrated that long noncoding RNAs (lncRNAs) are promising targets for the early diagnosis, prognosis and treatment of AML. A critical review of available data would be important to identify study gaps and provide perspectives. In this review, we explored comprehensive information on the potential use of lncRNAs as targets for the diagnosis, prognosis, and treatment of AML. LncRNAs are nonprotein-coding RNAs that are approximately 200 nucleotides long and play important roles in the regulation, metabolism and differentiation of tissues. In addition, they play important roles in the diagnosis, prognosis and treatment of different cancers, including AML. LncRNAs play multifaceted roles as oncogenes or tumor suppressor genes. Recently, deregulated lncRNAs were identified as novel players in the development of AML, making them promising prognostic indicators. Given that lncRNAs could have potential diagnostic marker roles, the lack of sufficient evidence identifying specific lncRNAs expressed in specific cancers hampers the use of lncRNAs as diagnostic markers of AML. The complex roles of lncRNAs in the pathophysiology of AML require further scrutiny to identify specific lncRNAs. This review, despite the lack of sufficient literature, discusses the therapeutic, diagnostic and prognostic roles of lncRNAs in AML and provides future insights that will contribute to studies targeting lncRNAs in the diagnosis, treatment, and management of AML.

IgNAR antibody: Structural features, diversity and applications.

In response to the invasion of exogenous microorganisms, one of the defence strategies of the immune system is to produce antibodies. Cartilaginous fish is among those who evolved the earliest humoral immune system that utilizes immunoglobulin-type antibodies. The cartilaginous fish antibodies fall into three categories: IgW, IgM, and IgNAR. The shark Immunoglobulin Novel Antigen Receptor (IgNAR) constitutes disulfide-bonded dimers of two protein chains, similar to the heavy chain of mammalian IgGs. Shark IgNAR is the primary antibody of a shark's adaptive immune system with a serum concentration of 0.1-1.0 mg/mL. Its structure comprises of one variable (V) domain (VNAR) and five constant (C1 -C5) domains in the secretory form. VNARs are classified into several subclasses based on specific properties such as the quantity and position of additional non-canonical cysteine (Cys) residues in the VNAR. The VDJ recombination in IgNAR comprises various fragments; one variable component, three diverse sections, one joining portion, and a solitary arrangement of constant fragments framed in each IgNAR gene cluster. The re-arrangement happens just inside this gene cluster bringing about a VD1D2D3J segment. Therefore, four re-arrangement procedures create the entire VNAR space. IgNAR antibody can serve as an excellent diagnostic, therapeutic, and research tool because it has a smaller size, high specificity for antigen-binding, and perfect stability. The domain characterization, structural features, types, diversity and therapeutic applications of IgNAR molecules are highlighted in this review. It would be helpful for further research on IgNAR antibodies acting as an essential constituent of the adaptive immune system and a potential therapeutic agent.

Diagnostic accuracy of serological tests and kinetics of severe acute respiratory syndrome coronavirus 2 antibody: A systematic review and meta-analysis.

This study aimed to assess the diagnostic test accuracy (DTA) of severe acute respiratory syndrome coronavirus 2 (SARS-COV-2) serological test methods and the kinetics of antibody positivity. Systematic review and meta-analysis were conducted following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses guideline. We included articles evaluating the diagnostic accuracy of serological tests and the kinetics of antibody positivity. MEDLINE through PubMed, Scopus, medRxiv and bioRxiv were sources of articles. Methodological qualities of included articles were appraised using QUADAS-2 while Metandi performs bivariate meta-analysis of DTA using a generalized linear mixed-model approach. Stata 14 and Review Manager 5.3 were used for data analysis. The summary sensitivity/specificity of chemiluminescence immunoassay (CLIA), enzyme-linked immunosorbent assay (ELISA) and lateral flow immunoassay (LFIA) were 92% (95% CI: 86%-95%)/99% (CI: 97%-99%), 86% (CI: 82%-89%)/99% (CI: 98%-100%) and 78% (CI: 71%-83%)/98% (95% CI: 96%-99%), respectively. Moreover, CLIA-based assays produced nearly 100% sensitivity within 11-15 days post-symptom onset (DPSO). Based on antibody type, the sensitivity of ELISA-total antibody, CLIA-IgM/G and CLIA-IgG gauged at 94%, 92% and 92%, respectively. The sensitivity of CLIA-RBD assay reached 96%, while LFIA-S demonstrated the lowest sensitivity, 71% (95% CI: 58%-80%). CLIA assays targeting antibodies against RBD considered the best DTA. The antibody positivity rate increased corresponding with DPSO, but there was some decrement when moving from acute phase to convalescent phase of infection. As immunoglobulin isotope-related DTA was heterogeneous, our data have insufficient evidence to recommend CLIA/ELISA for clinical decision-making, but likely to have comparative advantage over RT-qPCR in certain circumstances and geographic regions.

Aging, inflammaging and immunosenescence as risk factors of severe COVID-19.

Coronavirus disease 2019 (COVID-19) is a respiratory infectious disease caused by the novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). COVID-19 is characterized by having a heterogeneous disease course, ranging from asymptomatic and mild symptoms to more severe and critical cases. In most cases the severity of COVID-19 is related to host factors, especially deregulation of the immune response in patients. Even if COVID-19 indiscriminately affects individuals of different age group, ethnicity and economic status; most severe cases and disproportional mortality occur in elderly individuals. This point out that aging is one risk factor for unfavourable clinical outcomes among COVID-19 patients. The biology of aging is a complex process; Aging can alter the structure and function of cells, tissues, and organs resulting in impaired response to stress. Alongside with other systems, the immune system is also affected with the aging process. Immunosenescence is an age associated change in the immune system that affects the overall response to immunological challenges in the elderly. Similarly, apart from the normal inflammatory process, aging is associated with a low grade, sterile, chronic inflammation which is termed as inflammaging. We hypothesized that inflammaging and immunosenescence could play an important role in SARS-CoV-2 pathogenesis and poor recovery from COVID-19 in elderly individuals. This review summarizes the changes in the immune system with age and how these changes play part in the pathogenesis of SARS-CoV-2 and clinical outcome of COVID-19 which could add to the understanding of age associated targeted immunotherapy in the elderly.

Generation, biochemical characterizations and validation of potent nanobodies derived from alpaca specific for human receptor of advanced glycation end product.

A detrimental role of the receptor for the advanced glycation end product (RAGE) has been identified in the immune response, and various pathological conditions and its V and C1 domains in the extracellular region of RAGE are believed to be the main ligand-binding domains. Consequently, specific inhibitors targeting those domains could be of clinical value in fighting against the pathological condition associated with RAGE over-activation. Single-domain antibodies, also called nanobodies (Nbs), are antibody fragments engineered from the heavy-chain only antibodies found in camelids, which offer a range of advantages in therapy. In this study, we report the development and characterization of the V-C1 domain-specific Nbs. Three Nbs (3CNB, 4BNB, and 5ENB) targeting V-C1 domain of human RAGE were isolated from an immunized alpaca using a phage display. All of these Nbs revealed high thermostability. 3CNB, 4BNB, and 5ENB bind to V-C1 domain with a dissociation constant (KD) of 27.25, 39.37, and 47.85 nM, respectively, using Isothermal Titration Calorimetry (ITC). After homodimerization using human IgG1-Fc fusion, their binding affinity improved to 0.55, 0.62, and 0.41 nM, respectively, using Surface Plasmon Resonance (SPR). Flow cytometry showed all the Fc fusions Nbs can bind to human RAGE expressed on the cell surface. Competitive ELISA further confirmed their V-C1-hS100B blocking ability in solution, providing insights into the applicability of Nbs in treating RAGE-associated diseases.

The COVID-19 Severity and Its Association with Intestinal Parasite Coinfection and Urine Biochemical Parameters among COVID-19-Confirmed Patients Admitted to Debre Markos University COVID-19 Center, Northwest Ethiopia.

Background: Though most people with COVID-19 disease show asymptomatic to mild illness, a substantial number of patients are at high risk of developing severe disease and adverse outcomes with long COVID-19 and death. Even though some studies showed that previously existing infections with parasites amend the host's body defenses to increase resistance to infection with SARS-CoV-2, there is limited data in Ethiopia. Objectives: This study is aimed at determining the COVID-19 disease severity and its association with intestinal parasite coinfection and urine biochemical parameters among COVID-19-confirmed patients admitted at Debre Markos University COVID-19 Center, 2021. Methods: A prospective cohort study was conducted on 136 RT-qPCR-confirmed COVID-19 patients admitted at Debre Markos University COVID-19 Center from January 1 to March 30, 2021. Sociodemographic and clinical data were collected by using standardized data collection forms. A urine biochemical test was performed using a dry urine dipstick kit and stool examination using direct wet mount microscopic examination and formalin-ether concentration method. The chi-square test, Fisher exact test, and ordinal logistic regression analysis were computed to assess association with outcome variables using Statistical Package for Social Science software (version 24). Result: A total of 136 COVID-19-confirmed patients participated in this study. The median age of the participants was 48 years. The majority (86 (62.5%)) of them were male in sex. Of the 136 cases, 39 (28.7%) had died. Among the 136 patients, 22 (16.2%) were coinfected with intestinal parasites. COVID-19 patients who have intestinal parasite coinfection had lower odds of developing clinically severe COVID-19 compared to noninfected (AOR = 0.37; 95% CI = 0.147-0.944; P = 0.037). The majority (104 (76.5%)) of them have abnormal urine biochemical results. From the abnormal urine biochemical tests observed, the urine blood, glucose, and ketone tests were positive for 54 (39.7%), 36 (26.5%), and 30 (21.1%) patients, respectively. Among the 31 critical COVID-19 patients, 25 (80.6%) showed abnormal urine biochemical parameters. Age and comorbidity were significantly associated with COVID-19 severity (P < 0.05). Conclusion: Patients with old age and comorbidity had an increased risk of developing severe COVID-19 disease. Patients having SARS-CoV-2 and intestinal parasitic coinfections demonstrated mild COVID-19 disease severity. Abnormal urine biochemical results were common among critical COVID-19 patients. Thus, advanced study on the effect of the interaction among intestinal parasites on COVID-19 clinical severity and its mechanisms is essential.

Prevalence, antimicrobial resistance pattern, and associated factors of Salmonella and Shigella among under five diarrheic children attending public health facilities in Debre Markos town, Northwest Ethiopia.

Background: Under five children are at risk of diarrhea-associated morbidity and mortality. Salmonella and Shigella are major causes of diarrhea in under-five children, especially in developing countries. This study aimed to assess the prevalence, antimicrobial resistance pattern, and associated factors of Salmonella and Shigella among under-five diarrheic children in Debre Markos town public health facilities. Methods: A cross-sectional study was conducted at public health facilities in Debre Markos town using a consecutive convenient sampling technique. Data on socio-demographic and associated factors were collected using a structured questionnaire. Salmonella serovars and Shigella species were identified using MacConkey, Xylose Lysine Deoxycholate, Salmonella Shigella agar, and biochemical tests. The antimicrobial resistance pattern was determined by using the modified Kirby-Bauer disk diffusion technique. Results: The overall prevalence of Salmonella and Shigella was 11.7% (26/222; 95% CI = 7.2-17.5%). Isolated Salmonella serovars showed a higher rate of resistance (85.7%, 6/7) for both Ampicillin and Amoxicillin/Clavulanic acid while Shigella isolates showed a higher resistance rate to Amoxicillin/Clavulanic acid (78.9%, 15/19) and Ampicillin (73.7%, 14/19). The overall multidrug resistance (MDR) rate of Salmonella and Shigella isolates was 88.5% (23/26). Parent/guardian educational status ≤ elementary school (AOR = 3.783; 95% CI = 1.28-11.19; P = 0.016), presence of two or more under-five children in the family (AOR = 8.999; 95% CI = 2.93-27.69; P < 0.001), unimproved source of drinking water (AOR = 5.010; 95% CI = 1.56-16.10; P = 0.007), the habit of storing cooked foods for later use (AOR = 3.199; 95% CI = 1.07-9.54; P = 0.037), attendance of the child at social gatherings (AOR = 5.387; 95% CI = 1.78-16.35; P = 0.003), and infrequent child fingernail trimming (every ≥ 2 weeks; AOR = 4.693; 95% CI = 1.47-14.94; P = 0.009) showed statistically significant association with the prevalence of culture-confirmed Salmonella and Shigella isolates. Conclusion: The prevalence of culture-confirmed Salmonella and Shigella isolates was significantly high in the study area. Salmonella and Shigella isolates exhibited a high rate of MDR pattern. Parent/guardian education level below the elementary school, the presence of two or more under-five children in the family, using unimproved water source, a habit of storing cooked food, and infrequent fingernail trimming were independent predictors of culture-confirmed Salmonella and Shigella. Therefore, besides public health measures, regular surveillance of the prevalence and antimicrobial resistance pattern of Salmonella and Shigella should be routinely practiced in the study setting.

In silico Screening of Potential SARS-CoV-2 Main Protease Inhibitors from Thymus schimperi.

Background: COVID-19 is still instigating significant social and economic chaos worldwide; however, there is no approved antiviral drug yet. Here, we used in silico analysis to screen potential SARS-CoV-2 main protease (Mpro) inhibitors extracted from the essential oil of Thymus schimperi which could contribute to the discovery of potent anti-SARS-CoV-2 phytochemicals. Methods: The absorption, distribution, metabolism, excretion, and toxicity (ADMET) profiles of compounds were determined through SwissADME and ProToxII servers. AutoDock tools were used for molecular docking analysis studies, while Chimera, DS studio, and LigPlot were used for post-docking studies. Molecular dynamic simulations were performed for 200 ns under constant pressure. Results: All compounds exhibited a bioavailability score of ≥0.55 entailing that at least 55% of the drugs can be absorbed unchanged. Only five (9%), nine (16%) and two (3.6%) of the compounds showed active hepatotoxicity, carcinogenicity, and immunotoxicity, respectively. Except for flourazophore P, which showed a little mutagenicity, all other compounds did not show mutagenic properties. On the other hand, only pinene beta was found to have a little cytotoxicity. Five compounds demonstrated effective binding to the catalytic dyad of the SARS-CoV-2 Mpro substrate binding pocket, while two of them (geranylisobutanoate and 3-octane) are found to be the best hits that formed hydrogen bonds with Glu166 and Ser144 of SARS-CoV-2 Mpro. Conclusion: Based on our in silico analysis, top hits from Thymus schimperi may serve as potential anti-SARS-CoV-2 compounds. Further in vitro and in vivo studies are recommended to characterize these compounds for clinical applications.

Bacterial isolates, their antimicrobial susceptibility pattern, and associated factors of external ocular infections among patients attending eye clinic at Debre Markos Comprehensive Specialized Hospital, Northwest Ethiopia.

BACKGROUND: External eye infection caused by bacteria can lead to reduced vision and blindness. Therefore, pathogen isolation and antimicrobial susceptibility testing are vital for the prevention and control of ocular diseases. OBJECTIVE: The main aim of this study was to assess bacterial isolates, their antimicrobial susceptibility pattern, and associated factors of external ocular infection (EOI) among patients attended eye clinic at Debre Markos Comprehensive Specialized Hospital (DMCSH), Northwest Ethiopia. METHODS: We conducted a cross-sectional study in patients with external ocular infections from January 1, 2021, to June 30, 2021, at DMCSH. Socio-demographic and clinical data were collected using semi-structured questionnaires. Following standard protocols, external ocular swabs were collected and inoculated onto blood agar, chocolate agar, MacConkey agar and mannitol salt agar (MSA). Finally, bacterial isolates were identified by Gram stain, colony morphology, and biochemical tests. Antimicrobial susceptibility testing was done by using the modified Kirby-Bauer disk diffusion technique according to Clinical and Laboratory Standards Institute (CLSI) guideline. Cleaned and coded data were entered into EpiData version 4.2 software and exported to Statistical Packages for Social Sciences (SPSS) version 22 for analysis. Bivariate logistic regression was applied to investigate the association between predictors and outcome variables. P-values ≤ 0.05 with 95% confidence interval were considered statistically significant. RESULTS: Two hundred seven study participants were enrolled in this study. More than half of them (57.5%, 119/207) were males, and 37.7% (78/207) of them were ≥ 65 years old. A total of 130 (62.8%) bacterial isolates were identified, with Gram-positive bacteria accounting for 78.5% (102/130) of the isolates. Staphylococcus aureus was the most common isolate with a 46.2% (60/130) prevalence. Ciprofloxacin was comparatively effective against Gram-positive and Gram-negative bacteria. The prevalence of culture-confirmed bacteria was significantly associated with age groups 15-24 (AOR: 9.18, 95%CI: 1.01-82.80; P = 0.049) and 25-64 (AOR: 7.47, 95%CI: 1.06-52.31; P = 0.043). Being farmer (AOR: 5.33, 95% CI: 1.04-37.33; P = 0.045), previous history of eye surgery (AOR: 5.39, 95% CI: 1.66-17.48; P = 0.005), less frequency of face washing (AOR: 5.32, 95% CI: 1.31-7.23; P = 0.010) and face washing once a day (AOR: 3.07, 95% CI: 1.13-25.13; P = 0.035) were also significantly associated with the prevalence of culture-confirmed bacteria. CONCLUSION: The prevalence of culture-confirmed bacteria among patients with EOI was high in the study area. A considerable proportion of bacterial isolates exhibited mono and/or multi-drug resistance. Age (15-64 years), being farmer, previous history of eye surgery and less frequency of face washing were significantly associated with the prevalence of culture-confirmed bacteria. Bacterial isolation and antibiotic susceptibility testing should be routinely performed in the study area to combat the emergence of antibiotic resistance.

Prevalence, Antibiogram, and Associated Factors of Bacteria Isolated From Presumptive Meningitis Patients at Debre Markos Comprehensive Specialized Hospital, Northwest Ethiopia.

OBJECTIVE: Bacterial meningitis (BM) is a public health threat with considerable mortality and morbidity worldwide; particularly in the meningitis belt of Africa where Ethiopia is located. The study aims to assess the prevalence, antibiogram, and associated factors of bacteria isolated from presumptive meningitis patients at Debre Markos Comprehensive Specialized Hospital (DMCSH), Northwest Ethiopia. METHODS: We conducted a cross-sectional study between March 1, 2021, and May 30, 2021. Socio-demographic and clinical data were collected using structured questionnaires. Cerebrospinal fluid (CSF) was collected aseptically, and gram stain, culture, and biochemical tests were performed to identify bacterial isolates. An antimicrobial susceptibility test was conducted using the disc diffusion method on Mueller-Hinton agar (MHA). Data were entered into EpiData version 3.1 (Epidata Association, Denmark) and exported to SPSS version 23 software (IBM Corp., Armonk, NY) for analysis. P values ≤ 0.05 at 95% CI were considered statistically significant. RESULTS: CSF samples from 152 study participants were analyzed and half (50%, 76/152) of them were males. Bacteria were isolated from 17 individuals with an overall prevalence rate of 11.2% (95% CI= 5.9-16.4). The predominant bacterial isolates were Staphylococcus aureus (S. aureus) and Klebsiella pneumonia (K. pneumoniae) each accounting for 29.4% (5/17). About 41% (7/17) of the isolated bacteria were found to be multi-drug resistant (MDR) with the predominance of gram-negative bacteria (6/7). Bacteria prevalence was significantly higher in individuals with stiff neck [adjusted odds ratio (AOR), 95% CI, 47.529 (3.2-10.92), P=0.023] and tonsillectomy [AOR, 95% CI, 137.015 (6.25-12.34), P=0.02]. CONCLUSION: S. aureus and K. pneumoniae were the leading isolates among presumptive meningitis patients. The alarming presence of a high rate of MDR isolates mandates the need to implement the antibiotic stewardship program in the study setting.

Nanotechnology: A Promising Approach for Cancer Diagnosis, Therapeutics and Theragnosis.

Cancer remains the most devastating disease and the major cause of mortality worldwide. Although early diagnosis and treatment are the key approach in fighting against cancer, the available conventional diagnostic and therapeutic methods are not efficient. Besides, ineffective cancer cell selectivity and toxicity of traditional chemotherapy remain the most significant challenge. These limitations entail the need for the development of both safe and effective cancer diagnosis and treatment options. Due to its robust application, nanotechnology could be a promising method for in-vivo imaging and detection of cancer cells and cancer biomarkers. Nanotechnology could provide a quick, safe, cost-effective, and efficient method for cancer management. It also provides simultaneous diagnosis and treatment of cancer using nano-theragnostic particles that facilitate early detection and selective destruction of cancer cells. Updated and recent discussions are important for selecting the best cancer diagnosis, treatment, and management options, and new insights on designing effective protocols are utmost important. This review discusses the application of nanotechnology in cancer diagnosis, therapeutics, and theragnosis and provides future perspectives in the field.

Phage Therapy: A Different Approach to Fight Bacterial Infections.

Phage therapy is one of the alternatives to treat infections caused by both antibiotic-sensitive and antibiotic-resistant bacteria, with no or low toxicity to patients. It was started a century ago, although rapidly growing bacterial antimicrobial resistance, resulting in high levels of morbidity, mortality, and financial cost, has initiated the revival of phage therapy. It involves the use of live lytic, bioengineered, phage-encoded biological products, in combination with chemical antibiotics to treat bacterial infections. Importantly, phages will be removed from the body within seven days of clearing an infection. They target specific bacterial strains and cause minimal disruption to the microbial balance in humans. Phages for medication must be screened for the absence of resistant genes, virulent genes, cytotoxicity, and their interaction with the host tissue and organs. Since they are immunogenic, applying a high phage titer for therapy exposes them and activates the host immune system. To date, no serious side effects have been reported with human phage therapy. In this review, we describe phage-phagocyte interaction, bacterial resistance to phages, how phages conquer bacterial resistance, the role of genetic engineering and other technologies in phage therapy, and the therapeutic application of modified phages and phage-encoded products. We also highlight the comparison of antibiotics and lytic phage therapy, the pros and cons of phage therapy, determinants of human phage therapy trials, phage quality and safety requirements, phage storage and handling, and current challenges in phage therapy.

SARS-CoV-2 subunit vaccine adjuvants and their signaling pathways.

INTRODUCTION: Vaccines are the agreed upon weapon against the COVID-19 pandemic. This review discusses about COVID-19 subunit vaccines adjuvants and their signaling pathways, which could provide a glimpse into the selection of appropriate adjuvants for prospective vaccine development studies. AREAS COVERED: In the introduction, a brief background about the SARS-CoV-2 pandemic, the vaccine development race and classes of vaccine adjuvants were provided. . The antigen, trial stage, and types of adjuvants were extracted from the included articles and thun assimilated. Finally, the pattern recognition receptors (PRRs), their classes, cognate adjuvants, and potential signaling pathways were comprehended. EXPERT OPINION: Adjuvants are unsung heroes of subunit vaccines. The in silico studies are very vital in avoiding several costly trial errors and save much work times. The majority of the (pre)clinical studies are promising. It is encouraging that most of the selected adjuvants are novel. Much emphasis must be paid to the optimal paring of antigen-adjuvant-PRRs for obtaining the desired vaccine effect. A good subunit vaccine/adjuvant is one that has high efficacy, safety, dose sparing, and rapid seroconversion rate and broad spectrum of immune response. In the years to come, COVID-19 adjuvanted subunit vaccines are expected to have superior utility than any other vaccines for various reasons.

Prevalence of Medication Non-Adherence and Associated Factors among Diabetic Patients in A Tertiary Hospital at Debre Markos, Northwest Ethiopia.

Background: Non-adherence to prescribed medications is possibly the most common reason for poor treatment outcomes among people with diabetes although its rate is highly variable. Data on the magnitude of medication non-adherence and associated factors are scarce in the study area. This study aimed to assess the rate of non-adherence and associated factors among diabetic patients at Debre Markos Comprehensive Specialized Hospital. Methods: A cross-sectional study was conducted from June 17 to July 17, 2021. Study participants were selected using a simple random sampling technique. Data were collected with a pre-tested structured questionnaire and entered into SPSS version 25. Logistic regression was utilized to determine predictors of medication non-adherence at a significance level of ≤ 0.05. Results: A total of 176 study participants were enrolled in the study. About 59% of the study participants had type-2 diabetes mellitus. The prevalence of non-adherence to anti-diabetic medications was found to be 41.5%. Male sex, rural residence, being divorced, being merchant, self- or family-borne medical cost, and presence of comorbidities were significantly associated with increased rate of non-adherence to anti-diabetic medications. Conclusion: The prevalence of non-adherence to medications among diabetic patients is significantly high in the study area. Public health measures should be strengthened to decrease nonadherence among diabetic patients.

Structural Basis of Potential Inhibitors Targeting SARS-CoV-2 Main Protease.

The Coronavirus disease-19 (COVID-19) pandemic is still devastating the world causing significant social, economic, and political chaos. Corresponding to the absence of globally approved antiviral drugs for treatment and vaccines for controlling the pandemic, the number of cases and/or mortalities are still rising. Current patient management relies on supportive treatment and the use of repurposed drugs as an indispensable option. Of a crucial role in the viral life cycle, ongoing studies are looking for potential inhibitors to the main protease (Mpro) of severe acute respiratory syndrome Coronavirus -2 (SARS-CoV-2) to tackle the pandemic. Although promising results have been achieved in searching for drugs inhibiting the Mpro, work remains to be done on designing structure-based improved drugs. This review discusses the structural basis of potential inhibitors targeting SARS-CoV-2 Mpro, identifies gaps, and provides future directions. Further, compounds with potential Mpro based antiviral activity are highlighted.

Epidemiological, Clinical and Laboratory Characteristics of Patients with Brucella Infection in Anhui Province, China.

BACKGROUND: Brucellosis is currently one of the most widespread zoonotic diseases caused by Brucella genus, and the Brucella melitensis is the major pathogen. The number of people infected with Brucella has gradually increased in Anhui Province. PURPOSE: To retrospectively evaluate the epidemiological, clinical, and laboratory data of brucellosis patients in Anhui Province. PATIENTS AND METHODS: A total of 109 brucellosis patients were admitted to the First Affiliated Hospital of Anhui Medical University from January 2012 to March 2021. Data from all patients were retrieved from the hospital's electronic medical system. The final results were grouped and compared according to the presence or absence of bacteremic brucellosis and three phases of brucellosis. RESULTS: The most common symptoms among all 109 brucellosis patients were fever (89.0%), followed by chills (52.3%), arthralgia (48.6%), and weight loss (30.3%), and laboratory results presented with anemia (65.1%), elevate of C-reactive protein (CRP) (91.7%), erythrocyte sedimentation rate (ESR) (86.2%), aspartate aminotransferase (AST) (40.4%), and lactate dehydrogenase (LDH) (43.1%). The percentage of fever (96.1%), arthralgia (58.8%), anorexia (35.3%), leukopenia (31.4%), and the AST (51.0%) were higher in bacteremic than nonbacteremic group. Additionally, the median level of LDH (332.0 mg/L, IQR, 209.0-553.0) was higher in bacteremic than nonbacteremic group. Nevertheless, the albumin (36.0 mg/L, IQR, 33.9-38.2) was lower in the bacteremic group. The percentage of fever (94.9%) and the median LDH level (316.0 U/L (IQR,218.0-517.5)) in the acute phase of brucellosis were higher than the percentage of fever (72.0%) and the median LDH level (209.0 U/L (IQR,162.0-276.0)) in the subacute phase of brucellosis. CONCLUSION: Brucellosis has become an important public health issue in Anhui Province. Brucellosis is a disease with diverse clinical manifestations. Our data showed that unexplained fever, arthralgia, and elevated AST and LDH should be considered as a diagnosis of bacteremia brucellosis for early treatment intervention.

Effects of intestinal parasite infection on hematological profiles of pregnant women attending antenatal care at Debre Markos Referral Hospital, Northwest Ethiopia: Institution based prospective cohort study.

BACKGROUND: Intestinal parasitosis is a common disease that causes misery and disability in poor populations. The number of individuals affected is staggering. From two billion peoples who harbor parasites worldwide, 300 million suffer severe morbidity and more than 25% of pregnant women are infected with hookworm, which causes intestinal bleeding and blood loss, and has been most commonly associated with anemia. Intestinal parasite infection during pregnancy has been associated with iron deficiency, maternal anemia, and impaired nutritional status, as well as decreased infant birth weight. OBJECTIVE: This study aimed to assess the effects of intestinal parasite infection on hematological profiles of pregnant women attending antenatal care in Debre Markos Referral Hospital from December 2017 to February 2019. METHOD: A prospective cohort study design was conducted among 94 intestinal parasite-infected pregnant women as an exposed group and 187 pregnant women free from intestinal parasite were used as a control group. The effect of intestinal parasites on hematological profiles of pregnant women was assessed at Debre Markos Referral Hospital antenatal care ward. Socio-demographic data and nutrition status were assessed by using structured questionnaires and mid-upper arm circumference (MUAC), respectively. Two ml of venous blood and 2 gm of stool samples were collected to analyze the hematological profiles and detect intestinal parasites, respectively. Wet mount and formol-ether concentration (FEC) techniques were used to detect intestinal parasites. Hematological profile was analyzed using Mind ray BC-3000 plus instrument. Data were double entered into EpiData version 3.1 software and exported to SPSS version 24 software for analysis. Results were presented using tables and graphs. Associations of hemoglobin levels with intestinal parasitic infections were determined using binary logistic regression models. P≤0.05 was considered statistically significant. The mean hematological profile difference between parasite-infected and parasite-free pregnant women was computed using independent t-test. RESULTS: In the present study, the predominant parasites identified were Entamoeba histolytica, hookworm, Giardia lamblia, Schistosoma mansoni, and Ascaris lumbricoides. About 8.2% of intestinal parasite-infected pregnant women had mild anemia while 4% had moderate anemia. Only 1.2% of intestinal parasite-free pregnant women developed moderate anemia. The mean HGB, HCT, MCV, MCH, and MCHC values of intestinal parasite-infected pregnant women were 12.8g/dl, 38.2%, 94.7fl, 33.1pg and 34.7g/dl, respectively. But the mean HGB, HCT, MCV, MCH and MCHC values of pregnant women who were free from intestinal parasites were 14.4 g/dl, 39.8%, 94.9fl, 33.9pg and 35.5g/dl, respectively. Anemia was strongly associated with hookworm (AOR = 21.29, 95%CI: 8.28-54.75, P<0.001), S.mansoni (AOR = 63.73, 95% CI: 19.15-212, P<0.001) and A.lumbricoide (AOR = 14.12, 95% CI 3.28-60.65, P<0.001). CONCLUSION: Intestinal parasitic infection in pregnant women caused adverse impact on hematological profiles and was an independent predictor of anemia. Intestinal parasitic infection significantly decreased pregnant the level of HGB, HCT, MCV, MCH, and MCHC values. To minimize maternal anemia deworming could be good before pregnancy.

Structural Basis of the Pore-Forming Toxin/Membrane Interaction.

With the rapid growth of antibiotic-resistant bacteria, it is urgent to develop alternative therapeutic strategies. Pore-forming toxins (PFTs) belong to the largest family of virulence factors of many pathogenic bacteria and constitute the most characterized classes of pore-forming proteins (PFPs). Recent studies revealed the structural basis of several PFTs, both as soluble monomers, and transmembrane oligomers. Upon interacting with host cells, the soluble monomer of bacterial PFTs assembles into transmembrane oligomeric complexes that insert into membranes and affect target cell-membrane permeability, leading to diverse cellular responses and outcomes. Herein we have reviewed the structural basis of pore formation and interaction of PFTs with the host cell membrane, which could add valuable contributions in comprehensive understanding of PFTs and searching for novel therapeutic strategies targeting PFTs and interaction with host receptors in the fight of bacterial antibiotic-resistance.

Potency, Safety, and Pharmacokinetic Profiles of Potential Inhibitors Targeting SARS-CoV-2 Main Protease.

Effective, safe, and pharmacokinetically suitable drugs are urgently needed to curb the ongoing COVID-19 pandemic. The main protease or 3C-like protease (Mpro or 3CLpro) of SARS-CoV-2 is considered an important target to formulate potent drugs corresponding to its crucial role in virus replication and maturation in addition to its relatively conserved active site. Promising baseline data on the potency and safety of drugs targeting SARS-CoV-2 Mpro are currently available. However, preclinical and clinical data on the pharmacokinetic profiles of these drugs are very limited. This review discusses the potency, safety, and pharmacokinetic profiles of potential inhibitors of SARS-CoV-2 Mpro and forward directions on the development of future studies focusing on COVID-19 therapeutics.