Infectious Disorders - Drug Targets (Formerly Current Drug Targets - Infectious Disorders) - Volume 21, Issue 4, 2021

Ocular tissues can serve as a reservoir for the SARS-CoV-2 virus which can not only cause conjunctivitis but also serve as a source of infection transmission to others. Additionally, the eye and its tear drainage apparatus can track the SARS-CoV-2 from the eye into the respiratory tract of the patient. The potential ocular presence of the SARS-CoV-2 in the eye of a patient can target ACE2 receptors in the endothelium of the conjunctival vessels and use the lacrimal sac a potential space to evade immune detection and clinical isolation. The recently reported case of COVID-19 after the acquisition of SARS-CoV-2 from a COVID-19 patient should alert the healthcare professionals dealing with COVID-19 patients that wearing masks alone cannot guarantee protection against infection transmission. Further studies, like isolation of SARS-CoV-2 from the eyes of patients with COVID-19, are needed to identify the eyes as a potential source of SARS-CoV-2 infection transmission.

Background: As countries and industries continue to cope with the unparalleled challenges presented by the novel coronavirus (COVID-19), a specific area of concern has been the uncertainty surrounding the impact of the COVID-19 pandemic on the global and Indian supply chains of the pharmaceutical industry. The COVID-19 crisis has demonstrated the importance of establishing a risk management system that focuses on assessing future risks resulting from the loss of a supply chain among countries. Objective: This review focuses on the role of the Indian pharmaceutical industry towards the pandemic. This review investigates the economic effect of COVID-19 across segments and what it implies for the Indian economy. Methods: The COVID 19 flare-up has additionally commenced the Indian pharmaceutical organizations an opportunity to transform into a supported trade place point for gathering drugs and intermediates. Results: An enormous pharmaceutical industry in India has consistently been a foundation of reasonable human services, and this pattern would now be able to be required to heighten further. Conclusion: The activities from COVID-19 are with a need to change the overall impression of Indian pharmaceutical associations and even more altogether, reduce the dependence of the private pharma associations on alone suppliers like China.

Fever is a response of a human body, due to an increase in the temperature, against certain stimuli. It may be associated with several reasons and one of the major causes of fever is a mosquito bite. Fever due to dengue virus (DENV) infection is being paid most attention out of several other fever types because of a large number of deaths reported worldwide. Dengue virus is transmitted by biting of the mosquitoes, Aedes aegypti and Aedes albopictus. DENV1, DENV2, DENV3 and DENV4 are the four serotypes of dengue virus and these serotypes have 65% similarities in their genomic structure. The genome of DENV is composed of single-stranded RNA and it encodes for the polyprotein. Structural and non-structural proteins (nsP) are the two major parts of polyprotein. Researchers have paid high attention to the non-structural protease (nsP) of DENV like nsP1, nsP2A, nsP2B, nsP3, nsP4A, nsP4B and nsP5. The NS2B-NS3 protease of DENV is the prime target of the researchers as it is responsible for the catalytic activity. In the present time, Dengvaxia (vaccine) is being recommended to patients suffering severely from DENV infection in few countries only. Till date, neither a vaccine nor an effective medicine is available to combat all four serotypes. This review describes the fever, its causes, and studies to cure the infection due to DENV using theoretical and experimental approaches.

The pandemic spread of COVID 19 caused by the novel Coronavirus (SARS-CoV- 2) produced a tremendous effect on the life of humanity across the globe. The epidemiological studies revealed the drastic spectrum of SARS-CoV 2 infection ranging from mere flu-like symptoms to severe respiratory suppression within a short period. Initially, cases have been confined in the emerging point, Wuhan, China. But, within a few months, it has spread all over 212 countries around the globe and presently has become a severe threat to human life. Even though it is a severe acute respiratory syndrome virus, recent reports came with multiple organ effects of SARS-CoV 2, suggesting the virulence potential of this novel virus to sweep the planet in the absence of a proper vaccine or therapy. In this review, we discuss the multi-organ pathophysiology of COVID-19 infection, together with the treatment methods adopted and innovative diagnostic methods used.

Background: Leishmaniasis is a public health problem and endemic in countries of the tropics and subtropics. An ongoing project with naked LACK (Leishmania homolog of receptors for activated C-kinase) demonstrated that this case of the gene is entirely susceptible to immune response and it does enter the cells effectively. This study aimed at developing a procedure to prepare a type of lipid nanoparticles overloaded with plasmid LACK (pcLACK) for usage as Leishmania major (L. major) nanoliposomal vaccine. Materials and Methods: The single-gene expression plasmid of pcLACK was encoded in the LACK antigen. Nanoparticles were set up by thin film procedure using cationic lipids 1, 2-Dioleoyl- 3-Trimethylammonium propane (DOTAP), 1, 2-Dioleoyl-snGlycero-3-Phosphoethanolamine (DOPE), and cholesterol in a molar proportion of 2:1:1 molar ratio. Using dynamic light scattering, the particle diameters of empty and loaded lipoplexes were measured in triplicate. The zeta-potential (ζ) was measured with the same instrument using the zeta potential mode as the average of 20 measurements by diluting the particles into a low salt buffer. Results: The results of the sustainability studies of Liposome-pcLACK formulation showed that there were no significant physical changes up to the 30th day of stability study at the storage condition of 4°C. However, there were significant changes in the formulation content during storage at 25°C for 30 days (204.2±0.90 at Day 30 compared with 207.2±0.26 nm at Day 0). It was observed that the prepared nanoliposomal formulation had more stability under refrigeration. Conclusion: Immunostimulatory cationic lipids bearing a pcLACK encapsulation could serve as an effective delivery system.

Background: Tobacco mosaic virus (TMV) is the most known virus in the plant mosaic virus family and is able to infect a wide range of crops, in particular, tobacco, causing a production loss. Objectives: Herein, and for the first time in Lebanon, we investigated the presence of TMV infection in crops by analyzing 88 samples of tobacco, tomato, cucumber and pepper collected from different regions in North Lebanon. Methods: Double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA), revealed a potential TMV infection of four tobacco samples out of 88 crop samples collected. However, no tomato, cucumber and pepper samples were infected. The TMV+ tobacco samples were then extensively analyzed by RT-PCR to detect viral RNA using different primers covering all the viral genome. Results and Discussion: PCR results confirmed those of DAS-ELISA showing TMV infection of four tobacco samples collected from three crop fields of North Lebanon. In only one of four TMV+ samples, we were able to amplify almost all the regions of viral genome, suggesting possible mutations in the virus genome or an infection with a new, not yet identified, TMV strain. Conclusion: Our study is the first in Lebanon, revealing TMV infection in crop fields and highlighting the danger that may affect the future of agriculture.

Background: Since December 2019, a novel coronavirus, SARS-CoV-2, has caused global public health issues after being reported for the first time in Wuhan province of China. So far, there have been approximately 14.8 million confirmed cases and 0.614 million deaths due to the SARS-CoV-2 infection globally, and still, numbers are increasing. Although the virus has caused a global public health concern, no effective treatment has been developed. Objective: One of the strategies to combat the COVID-19 disease caused by SARS-CoV-2 is the development of vaccines that can make humans immune to these infections. Considering this approach, in this study, an attempt has been made to design epitope-based vaccine for combatting COVID-19 disease by analyzing the complete proteome of the virus by using immuno-informatics tools. Methods: The protein sequence of the SARS-CoV-2 was retrieved and the individual proteins were checked for their allergic potential. Then, from non-allergen proteins, antigenic epitopes were identified that could bind with MHCII molecules. The epitopes were modeled and docked to predict the interaction with MHCII molecules. The stability of the epitope-MHCII complex was further analyzed by performing a molecular dynamics simulation study. The selected vaccine candidates were also analyzed for their global population coverage and conservancy among SARS-related coronavirus species. Results: The study has predicted 5 peptide molecules that can act as potential candidates for epitope- based vaccine development. Among the 5 selected epitopes, the peptide LRARSVSPK can be the most potent epitope because of its high geometric shape complementarity score, low ACE and very high response towards it by the world population (81.81% global population coverage). Further, molecular dynamic simulation analysis indicated the formation of a stable epitope-MHCII complex. The epitope LRARSVSPK was also found to be highly conserved among the SARS-CoV- -2 isolated from different countries. Conclusion: The study has predicted T-cell epitopes that can elicit a robust immune response in the global human population and act as potential vaccine candidates. However, the ability of these epitopes to act as vaccine candidate needs to be validated in wet lab studies.

Background: Methicillin-resistant Staphylococcus aureus (MRSA) can cause serious infections not only in hospitals but also in the community. The present study was aimed to characterize drug resistance and virulence determinants of community-associated (CA) MRSA isolate compared with healthcare-associated (HA) MRSA. Materials and Methods: A total of 44 patients with HA-MRSA and 11 patients with CA-MRSA infection (median age, 72 years) were included. The clinical isolates of MRSA were subjected to molecular analysis of virulence genes and drug susceptibility testing. Results: Panton-Valentine leucocidin (PVL) exotoxin and toxic shock syndrome toxin (TSST) genes were disproportionately distributed between CA- and HA-isolates. PVL genes were more likely to be found among CA-isolates (36.4%) than HA-isolates (18.2). TSST genes were identified in only 2 CA-MRSA isolates tested (18.2%) compared with 9 HA-isolates (20.5%). Exfoliative toxin- b gene was negative in all isolates, however, one HA-isolate was positive for exfoliative toxin-a. mec-A gene was present in all clinical isolates. CA-isolates were more likely to be susceptible to trimethoprim-sulfamethoxazole and vancomycin compared with HA-isolates. Vancomycin-intermediate resistance was found in 2 HA-isolates. All clinical isolates were also resistant to clindamycin. Conclusion: CA- and HA- MRSA isolates are epidemiologically and microbiologically distinct. Thus, the strategies to prevent and treat these infections would be different. Patients with CA- and HA-MRSA infections should be treated effectively and receive follow-up evaluation to ensure the resolution of their infection. Surveillance studies should be conducted to determine the extent of CA- and HA-MRSA dissemination in Iran.

Background: Drug- and multidrug-resistant Klebsiella pneumoniae isolates have been found worldwide. Treatment failures against carbapenems and extended-spectrum cephalosporins, the currently recommended drugs, contribute to consider K. pneumoniae infections as untreatable infections. The emergence and spread of oxacillinases (OXAs) with carbapenem-hydrolyzing properties are a major concern and seriously become a public health problem worldwide. The present study was aimed to explore the blaOXA genes among clinical isolates of K. pneumoniae in some clinical settings in Tehran, Iran. Methods: A total of 90 K. pneumoniae isolates were collected from different clinical samples at hospitals in Tehran during the year 2016 and 2018. Antimicrobial susceptibility testing was performed on bacterial isolates using the Kirby-Bauer disc diffusion method on Mueller Hinton agar plates. PCR experiments were carried out to detect the presence of the blaOXA genes, including blaOXA- 1, blaOXA-2, blaOXA-4, blaOXA10, and blaOXA-48-like, using specific primers. Results: The antibiotics susceptibility results showed that 41% of the K. pneumoniae isolates were resistant to imipenem and meropenem. Resistance rates for cephalosporin agents, including cefpodoxime, ceftazidime, cefuroxime, cefotaxime, and cefepime, were measured as 72.3%, 67.8%, 67.7%, 65.5%, and 60%, respectively. In the present study, 51.1% of isolates were classified as multidrug-resistant K. pneumoniae strains. The molecular assays showed that 56.6% of isolates harbored blaOXA-2. In addition, blaOXA-4, blaOXA-1, blaOXA-10, and blaOXA-48-like genes were also found in 16.7%, 5.6%, 1.1%, and 1.1% of isolates, respectively. Conclusion: The spread of blaOXAs, especially blaOXA-48-like, among K. pneumoniae isolates indicated the inadequate dissemination control of multidrug-resistant bacteria in the Iranian hospital environment. There is a reason to assume that OXA producing K. pneumoniae will limit clinical therapeutic options in the future and pose threats to national public health among the Iranian population.

Background: Increasing prevalence of multiple antibiotic resistance in Klebsiella pneumoniae strains confines the therapeutic options used to treat bacterial infections. Objective: We aimed in this study to investigate the role of AcrAB and qepA efflux pumps and AAC(6′)-Ib-cr enzyme in ciprofloxacin resistance and to detect the RAPD-PCR fingerprint of K. pneumoniae isolates. Methods: A total of , 117 K. pneumoniae isolates were collected from hospitalized patients in three hospitals in Tehran, Iran, from August 2013 to March 2014. Antimicrobial susceptibility tests were performed by the disk diffusion method. Molecular identification and expression level of encoding quinolone resistance genes, acrA, acrB, qepA, and aac(6')-Ib-cr, were performed by PCR and real-- time PCR assays, respectively. All the K. pneumoniae isolates containing the mentioned genes were used simultaneously for RAPD-PCR typing. Results: Colistin and carbapenems were the most efficient antibiotics against the clinical isolates of K. pneumoniae. PCR assay demonstrated that among the 117 isolates, 110 (94%) and 102 (87%) were positive for acrA and acrB gene and 5 (4%) and 100 (85%) isolates showed to have qepA and aac(6′)-Ib-cr genes, respectively. Determination for AcrAB pump expression in 21% of strains demonstrated an increased expression, and the mean increase expression for acrB genes was 0.5-81. The results of RAPD-PCR reflected that in 95% CI, all isolates belonged to a clone. Conclusion: A high prevalence of genes encoding quinolone resistance in K. pneumoniae was detected in clinical samples. Therefore, the control of infection and prevention of drug-resistant bacteria spread need careful management of medication and identification of resistant isolates.

Background: Globally, the recent outbreak of Zika virus (ZIKV) in Brazil, Asia Pacific, and other countries highlighted the unmet medical needs. Currently, there are neither effective vaccines nor therapeutics available to prevent or treat ZIKV infection. Objective: In this study, we aimed to design an epitope-based vaccine for ZIKV using an in silico approach to predict and analyze B- and T-cell epitopes. Methods: The prediction of the most antigenic epitopes has targeted the capsid and envelope proteins as well as non-structural proteins NS5 and NS3 using immune-informatics tools PROTPARAM, CFSSP, PSIPRED, and Vaxijen v2.0. B and T-cell epitopes were predicted using ABCpred, IEDB, TepiTool, and their toxicity was evaluated using ToxinPred. The 3-dimensional epitope structures were generated by PEP-FOLD. Energy minimization was performed using Swiss- Pdb Viewer, and molecular docking was conducted using PatchDock and FireDock server. Results: As a result, we predicted 307 epitopes of MHCI (major histocompatibility complex class I) and 102 epitopes of MHCII (major histocompatibility complex class II). Based on immunogenicity and antigenicity scores, we identified the four most antigenic MHC I epitopes: MVLAILAFLR (HLA-A*68:01), ETLHGTVTV (HLA-A*68:02), DENHPYRTW (HLA-B*44:02), QEGVFH TMW (HLA-B*44:03) and TASGRVIEEW (HLA-B*58:01), and MHC II epitopes: IIKKFKKDLAAMLRI (HLA-DRB3*02:02), ENSKMMLELDPPFGD (HLA-DRB3*01:01), HAET WFFDENHPYRT (HLA-DRB3*01:01), TDGVYRVMTRRLLGS (HLA-DRB1*11:01), and DGCW YGMEIRPRKEP (HLA-DRB5*01:01). Conclusion: This study provides novel potential B cell and T cell epitopes to fight against Zika virus infections and may prompt further development of vaccines against ZIKV and other emerging infectious diseases. However, further investigations for protective immune response by in vitro and in vivo studies to ratify immunogenicity, the safety of the predicted structure, and ultimately for the vaccine properties to prevent ZIKV infections are warranted.

Background: Trichomoniasis is a sexually transmitted disease (STD) prevalent in different parts of the world. The present study was designed to evaluate the scientific output and collaboration of researchers worldwide on trichomoniasis. Methods: In the present scientometric research, all papers conducted on Trichomonas vaginalis (T. vaginalis) published in the time period of 1922 to 2018 were extracted from the Scopus database and evaluated according to the aims of the research, using scientometric software. Results: The results of this search strategy were the retrieval of 12,596 scientific papers authored by a total of 28,822 people during 97 years (1922-2018). Research in this field has encountered success and failure over the years. The first article on trichomoniasis at the Scopus database was published by Musgrave W.E. (1922) in the “Journal of the American Medical Association”. Most papers on this subject (352) were published in 2013; and the years 2017 and 2016 are ranked second and third. Researchers from the US Centers for Disease Control and Prevention have the highest number of papers in this field (210). Alderete J.F. with 111 papers is the most active researcher on trichomoniasis in Scopus. Also, researchers from the US ranked the first with 3,236 papers. In Iran, by the end of 2018, 138 papers on trichomoniasis have been registered in Scopus, which accounts for only one percent of the world's science production in this field. Discussion and conclusion: The results following about one century indicated that the number of publications on T. vaginalis is growing and that there is a high level of collaboration among researchers in this field. Moreover, many countries have collaborated internationally on trichomoniasis research with the US as the leading country.

Background: Extended Spectrum Beta Lactamase (ESBL) production in gram negative bacteria confers multiple antibiotic resistance, adversely affecting antimicrobial therapy in infected individuals. ESBLs result from mutations in β-lactamases encoded mainly by the blaTEM,blaSHV and blaCTX-M genes. The prevalence of ESBL producing bacteria has been on the increase globally, especially its upsurge among isolates from community-acquired infections has been observed. Aim: To determine ESBL prevalence and identify ESBL genes among clinical isolates in Osun State, Nigeria. Material and Methods: A cross-sectional study was carried out from August 2016 – July 2017 in Osun State, Nigeria. Three hundred and sixty Gram-negative bacteria recovered from clinical samples obtained from both community and healthcare-associated infections were tested. They included 147 Escherichia coli (40.8%), 116 Klebsiella spp (32.2%), 44 Pseudomonas aeruginosa (12.2%) and 23 Proteus vulgaris (6.4%) isolates. Others were Acinetobacter baumannii, Serratia rubidae, Citrobacter spp, Enterobacter spp and Salmonella typhi. Disk diffusion antibiotic susceptibility testing was carried out, isolates were screened for ESBL production and confirmed using standard laboratory procedures. ESBLs resistance genes were identified by Polymerase Chain Reaction (PCR). Results: All isolates demonstrated multiple antibiotic resistance. Resistance to ampicillin, amoxicillin with clavulanate and erythromycin was 100%, whereas resistance to Imipenem was very low (5.0%). The overall prevalence of ESBL producers was 41.4% with Klebsiella spp as the highest ESBL producing Enterobacteriacaea. ESBL producers were more prevalent among the hospital pathogens than community pathogens, 58% vs. 29.5% (p=0.003). ESBL genes were detected in all ESBL producers with the blaCTX-M gene predominating (47.0%) followed by blaTEM (30.9%) and blaSHV gene was the least, 22.1%. The blaCTX-M gene was also the most prevalent in the healthcare pathogens (62%) but it accounted for only 25% in those of community origin. Conclusion: A high prevalence of ESBL producing gram-negative organisms occurs both in healthcare and in the community in our environment with the CTX-M variant predominating. Efforts to control the spread of these pathogens should be addressed.

Introduction: Acute pyelonephritis is among the most common bacterial infections. Options for initial treatment of pyelonephritis include an extended-spectrum cephalosporin or a fluoroquinolone. This study aimed to compare the clinical outcomes of patients receiving ceftriaxone to those who received levofloxacin for the treatment of acute pyelonephritis. Methods: In this randomized, open-label trial, hospitalized adults with acute pyelonephritis were treated with ceftriaxone (1g IV every 12 hours) or levofloxacin (750 mg IV daily) for at least 7 days. Clinical and microbiological characteristics were compared among patients treated with ceftriaxone and levofloxacin. Results: A total of 59 patients were randomized, 30 to the ceftriaxone group and 29 to the levofloxacin group. The clinical response was that 68.0% of patients in the ceftriaxone group and 56.0% of patients in the levofloxacin group were cured. The microbiological response, i.e. pathogen eradication rates was 68.7% in the ceftriaxone group and 21.4% in the levofloxacin group. (P value=0.00028) Escherichia coli was the most common pathogen (n = 31), followed by Klebsiella pneumoniae (n = 21). High resistance rates were detected for cotrimoxazole (55%), ciprofloxacin (48%), and ceftriaxone (34.4%) in isolated E.coli. Likewise, all K. pneumoniae isolates were resistant to ciprofloxacin. Conclusion: This study indicates that ceftriaxone was more effective than levofloxacin in the treatment of acute pyelonephritis, on the basis of microbiological response, but there were no statistically significant differences between the treatment groups in the rates of clinical cure. The resistance of uropathogens to the most used antibiotics was relatively high. Choosing the treatment regimen based on susceptibility testing results and shortening the duration of the therapy are now recommended to be the most important approaches to decrease the spread of antibiotic resistance worldwide.

Background: COVID-19 is a life-threatening novel corona viral infection to our civilization and spreading rapidly. Tremendousefforts have been made by the researchers to search for a drug to control SARS-CoV-2. Methods: Here, a series of arsenical derivatives were optimized and analyzed with in silico study to search the inhibitor of RNA dependent RNA polymerase (RdRp), the major replication factor of SARS-CoV-2. All the optimized derivatives were blindly docked with RdRp of SARS-CoV-2 using iGEMDOCK v2.1. Results: Based on the lower idock score in the catalytic pocket of RdRp, darinaparsin (-82.52 kcal/- mol) was revealed to be the most effective among them. Darinaparsin strongly binds with both Nsp9 replicase protein (-8.77 kcal/mol) and Nsp15 endoribonuclease (-8.3 kcal/mol) of SARS-- CoV-2 as confirmed from the AutoDock analysis. During infection, the ssRNA of SARS-CoV-2 is translated into large polyproteins forming viral replication complex by specific proteases like 3CL protease and papain protease. This is also another target to control the virus infection where darinaparsin also performs the inhibitory role to proteases of 3CL protease (-7.69 kcal/mol) and papain protease (-8.43 kcal/mol). Conclusion: In the host cell, the furin protease serves as a gateway to the viral entry and darinaparsin docked with furin protease, which revealed a strong binding affinity. Thus, screening of potential arsenic drugs would help in providing the fast in-vitro to in-vivo analysis towards the development of therapeutics against SARS-CoV-2.

Introduction: Mycobacterium simiae is an emerging pathogen in Iran and little is known about drug susceptibility patterns of this pathogen. Materials and Methods: Twenty-five clinical isolates of M. simiae from 80 patients with confirmed NTM pulmonary disease were included in this study. For drug susceptibility testing (DST), proportional and broth microdilution methods were used according to the clinical and laboratory standards institute (CLSI) guideline. Results: All clinical isolates of M. simiae were resistant to isoniazid, rifampicin, ethambutol, streptomycin, amikacin, kanamycin, ciprofloxacin, and clarithromycin. They also were highly resistant to ofloxacin (80%). Susceptibility to ofloxacin was only noted in the 5 isolates. Conclusion: Clinical isolates of M. simiae were multidrug-resistant, and had different drug susceptibility patterns than previously published studies. DST results can assist in selecting more appropriate treatment regimens. Newer drugs with proven clinical efficacy correlating with in vitro susceptibility should be substituted with first- and second-line anti-TB drug testing.

Background and Aim: Colorectal Cancer (CRC) is one of the most frequent cancers diagnosed in both men and women worldwide. Fusobacterium nucleatum adhesin A (FadA) has an important potential factor in the development or progression of CRC. The aim of the present study was to evaluate the proliferative effect of recombinant FadA on SW480 colorectal cancer cell line. Materials and Methods: The recombinant pET21(b)-fadA plasmid was synthesized and transformed into competent E.coli DH5α. In the next step, induction and expression of recombinant FadA were carried out in E. coli BL21 (DE3) competent cells. Expression and purification of protein were successfully done and it was analyzed and confirmed by SDS-PAGE and western blotting. The proliferative effect of purified FadA on SW480 CRC cell line was evaluated using MTT assay and cell counting methods. Results: Visualization of the specific band isolated from the linear plasmid on the agarose gel confirmed the presence of the desired gene. After electrophoresis and Coomassie blue staining, the protein of interest with an approximate molecular weight of 13KDa was detected. The MTT assay, similar to cell counting methods, revealed that FadA dose and time-dependently promoted SW480 cell growth and proliferation in 24, 48 and 72 hours. Conclusion: The results showed that FadA stimulates proliferation of SW480 colorectal cancer cell line with a dose and time-dependent manner.