Search

Multidrug resistance patterns of Acinetobacter spp. have led to their emergence as an important source of nosocomial infections. This study investigated the prevalence and clinical characteristics of Acinetobacter spp. in hospital-acquired wound and urinary tract infections. A total of 432 samples [wound swabs (210) and urine samples (222)] were analyzed for the presence of Acinetobacter spp. through selective culturing on MacConkey and Leeds Acinetobacter medium followed by identification with API 20E strips and Vitek 2 compact system. Antimicrobial susceptibility was assessed by adopting the disk diffusion method on Muller-Hinton agar, whereas the minimum inhibitory concentration procedure was carried out by using a ComASP™ Colistin test kit. Biofilm formation was examined using microtiter plates and following the crystal violet staining method. PCR was performed to amplify virulence (lasB, bap, and plcN) and antimicrobial resistance (blaOXA-51like) genes. The results revealed a low prevalence of Acinetobacter spp. (1.85 %) where Acinetobacter baumannii was the predominant species. Acinetobacter baumannii isolates harbored blaOXA-51-like gene to exert extensive or pan-drug resistance. Most Acinetobacter baumannii isolates demonstrated weaker to moderate biofilm-forming capabilities and carried the bap gene. Acinetobacter baumannii isolates lacked the combination of virulence factors encoding lasB and plcN genes. Acinetobacter baumannii infections are rising in Saudi Arabia. The results of this study highlight the epidemiology of virulent and antibiotic-resistant Acinetobacter spp., particularly A. baumannii, in Saudi Arabia. The detailed elaboration on the diversity, virulence, and antimicrobial susceptibility patterns of Acinetobacter spp. in Saudi Arabia requires further in-depth molecular investigations.

The photoinitiated polymerization of 2-hydroxyethyl methacrylate (HEMA) (1–3 M) using camphorquinone (CQ) / triethanolamine (TEOHA) system has been studied in the pH range 6.0–9.0. The kinetics of the reaction has been evaluated during the initial stages causing about 5% HEMA conversion to avoid the effects of any variation in the volume of the solution. The change in the concentration of HEMA during polymerization has been determined by a UV spectrometric method at 208 nm with a precision of ±3%. The apparent first-order rate constants (kobs) for the polymerization of HEMA by CQ in the presence of 0.01 M TEOHA, carried out up to 100 s, range from 3.35–7.78 (1 M), 2.72–6.78 (2 M) and 2.33–5.89×10–4 s –1 (3 M). The second-order rate constants for the interaction of TEOHA radicals with HEMA (1–3 M) range from 2.33–7.78×10–2 M–1 s –1 at pH 6.0–9.0 indicating an increase in the reactivity of TEOHA radicals with an increase in the pH of the solution. There is a linear relation between kobs (1–3 M HEMA) and inverse of solution viscosity suggesting the quenching of CQ excited state with an increase in the viscosity of the medium

The kinetics of photolysis of ascorbic acid in aqueous solution on UV irradiation has been studied in the pH range 1–11 and the apparent first–order rate constants for the degradation reactions have been determined. The k–pH profile for the photolysis in the acid range is represented by a sigmoid curve indicating the gradual ionization of the molecule (AH2) to ascorbyl anion (AH– ). Ascorbic acid shows maximum stability around pH 5–6 due to the lowest rate of oxidation reduction of the mono–anion form. The rate of photolysis is increased up to sevenfold at pH 10.0, compared to that at pH 5.0, due to an increase in the redox potentials with pH leading to faster oxidation of the anionic species to dehydroascorbic acid in the alkaline range. The rate is very slow in the pH range 1–2 due to the existence of the non–ionized form. The apparent first–order rate constants for the photolysis of ascorbic acid at pH 1–11 range from 0.057–3.948×10–2 min–1 . A scheme for the sequence of reactions involved in the photolysis of ascorbic acid is presented.

Abstract Furin primarily localizes to the trans‐Golgi network (TGN), where it cleaves and activates a broad range of immature proproteins that play critical roles in cellular homeostasis, disease progression, and infection. Furin is retrieved from endosomes to the TGN after being phosphorylated, but it is still unclear how furin exits the TGN to initiate the post‐Golgi trafficking and how its activity is regulated in the TGN. Here three membrane‐associated RING‐CH finger (MARCHF) proteins (2, 8, 9) are identified as furin E3 ubiquitin ligases, which catalyze furin K33‐polyubiquitination. Polyubiquitination prevents furin from maturation by blocking its ectodomain cleavage inside cells but promotes its egress from the TGN and shedding. Further ubiquitin‐specific protease 32 (USP32) is identified as the furin deubiquitinase in the TGN that counteracts the MARCHF inhibitory activity on furin. Thus, the furin post‐Golgi trafficking is regulated by an interplay between polyubiquitination and phosphorylation. Polyubiquitination is required for furin anterograde transport but inhibits its proprotein convertase activity, and phosphorylation is required for furin retrograde transport to produce fully active furin inside cells.

Abstract SARS-CoV-2 and filovirus enter cells via the cell surface angiotensin-converting enzyme 2 (ACE2) or the late-endosome Niemann-Pick C1 (NPC1) as a receptor. Here, we screened 974 natural compounds and identified Tubeimosides I, II, and III as pan-coronavirus and filovirus entry inhibitors that target NPC1. Using in-silico, biochemical, and genomic approaches, we provide evidence that NPC1 also binds SARS-CoV-2 spike (S) protein on the receptor-binding domain (RBD), which is blocked by Tubeimosides. Importantly, NPC1 strongly promotes productive SARS-CoV-2 entry, which we propose is due to its influence on fusion in late endosomes. The Tubeimosides’ antiviral activity and NPC1 function are further confirmed by infection with SARS-CoV-2 variants of concern (VOC), SARS-CoV, and MERS-CoV. Thus, NPC1 is a critical entry co-factor for highly pathogenic human coronaviruses (HCoVs) in the late endosomes, and Tubeimosides hold promise as a new countermeasure for these HCoVs and filoviruses.

Purpose: Screening of Indian medicinal plants to obtain antioxidant plant extracts for their interference to bacterial quorum sensing and biofilm. Results: The most bioactive fraction of Moringa oleifera leaves extract, i.e., MOEF, has been identified and characterized in this study. In Chromobacterium violaceum 12472, MOEF significantly reduced the amount of violacein produced, i.e.,>70 %. Many virulence factors of Pseudomonas aeruginosa PAO1 were noticeably reduced in the presence of sub-MICs of MOEF. Adding MOEF to the culture media dramatically reduced prodigiosin, protease activity, and swarming of Serratia marcescens. With a >70 % decrease in biofilm development by the test pathogens, MOEF demonstrated broad-spectrum antibiofilm activity. SEM and CLSM examination showed that the biofilm growth on the glass coverslip was significantly decreased. GC/MS detected the major bioactive compounds 2-oleo-palmitostearin, 4-O-methylphorbol-12,13-didecanoate. The most numerous compounds of MOEF were identified using LC-qTOF-MS analysis. Further, the in vivo efficacy of active fraction needs to be assessed to uncover the therapeutic potential of MOEF. Conclusion: The findings of this study revealed that the most antioxidant fraction of M. oleifera, i.e., MOEF, demonstrated anti-infective efficacy in vitro against C. violaceum, P. aeruginosa, and S. marcescens. Concentration-dependent inhibition of Quorum-sensing regulated virulence factors and biofilms highlights its broad-spectrum action. It could be exploited in preparing standardized herbal bioactive products and developing modern phytomedicines. Further, the in vivo efficacy of active fraction needs to be assessed to uncover the therapeutic potential of MOEF.

Abstract Background: Tuberculosis affects millions of people across the globe annually. Pakistan is ranked 5th among the leading TB countries in the world. Anemia is a common observation among TB patients with the multifactorial phenomenon. The present study aimed to determine the frequency of anemia in patients with pulmonary TB and its classification as per the severity and morphological characteristics. Methodology: This retrospective cross-sectional study was conducted at DHQ Hospital Timergara, Lower Dir. A total of 109 TB patients were enrolled and underwent physical and systemic examination. Blood samples were collected to estimate complete blood counts (CBC), liver and renal function tests (LFT; RFT), serum albumin, and C-reactive protein (CRP) levels. Abdominal ultrasound was performed when any organomegaly was suspected. Hemoglobin was estimated using an automated analyzer (SYSMEX-SP100), and it was repeated via manual method among doubted cases. Written informed consent was obtained from all patients before inclusion, and the collected data was analyzed using SPSS version 22.0. Results: There was a female majority (54%) in the studied sample, with a mean age of 38 ± 15.3 years. Majority of them belonged to low socioeconomic status (80%). Moreover, 43% of them were from Dir Lower, 30% from Dir Upper, while the remaining were from Bajaur district, Chitral, and Malakand Swat districts. Anemia was found in 82.56% of the enrolled TB patients. Most patients were moderately weak (Hb < 10 g/dl > 7 g/dl), and microcytic anemia was found in more than 80% of the anemic patients. CRP level was raised in 70% of these patients, and there was hypoalbuminemia in 80%. Furthermore, females were more anemic than males. Conclusion: In our study, anemia was prevalent in TB patients, specifically microcytic anemia. Hence, TB patients must be monitored for dietary deficiencies and/or inflammatory modulators to avoid such complications.

Antimicrobial resistance (AMR) has become a critical global health challenge. Infections, particularly those caused by multidrug-resistant (MDR) pathogens, rank among the top causes of human mortality worldwide. Pseudomonas aeruginosa occupies a prominent position among pathogens responsible for opportunistic infections in humans. P. aeruginosa stands as a primary cause of chronic respiratory infections, significantly contributing to the burden of these chronic diseases. In the medical domain, nanotechnologies offer significant potential, spanning various applications, including advanced imaging, diagnostic devices, drug delivery systems, implants, tissue-engineered structures, and pharmaceutical treatments. Given the challenges associated with AMR and the limited discovery of new drugs to combat MDR microbes, there is a critical need for alternative strategies to address the problem of AMR. In this study, we synthesized titanium–cerium nanocomposites (Ti–Ce–NCs) using an eco-friendly green synthesis approach. X-ray diffraction (XRD) analysis confirmed the crystalline nature of the Ti–Ce–NCs and determined the particle size to be 17.07 nm. Electron microscopy revealed the size range of the particles to be 13 to 54 nm, where the majority of the particles were in the 20 to 25 nm range. Upon examining the composition, the Ti–Ce–NCs were determined to be composed of cerium, oxygen, and titanium, whose relative abundance were 36.86, 36.6, and 24.77% by weight, respectively. These nanocomposites were then evaluated for their effectiveness against various virulent traits and biofilms in P. aeruginosa. Out of six tested virulence factors, more than 50% inhibition of five virulence factors of P. aeruginosa was found. Roughly 60% inhibition of biofilm was also found in the presence of 400 µg/mL Ti–Ce–NCs. The nanocomposites also altered the biofilm architecture of the test bacterium. The success of this research opens doors for the potential use of such nanomaterials in the discovery of new antibacterial agents to combat drug-resistant bacteria.

Fixed point theory is a significant area of mathematical analysis with applications across various fields such as differential equations, optimization, and dynamical systems. Recently, multivalued mappings have gained attention due to their ability to model more complex and realistic problems. ln this work, novel classes of nonlinear mappings called k-strictly asymptotically demicontractive-type and asymptotically hemicontractive-type multivalued mappings are introduced in real Hilbert spaces that are symmetric spaces. In addition, we discuss the weak and strong convergence results by considered modified algorithms, and a demiclosedness property, for these classes of mappings are proved. Several non-trivial examples are demonstrated to validate the newly defined mappings. Consequently, the results and iterative methods obtained in this study improve and extend several known outcomes in the literature.

The main aim of this work is to use the XOR-operation technique to find the common solutions for a new class of extended system of fuzzy ordered variational inclusions with its corresponding system of fuzzy ordered resolvent equations involving the $ \oplus $ operation and fixed point problems, which are slightly different from corresponding problems considered in several recent papers in the literature and are more advantageous. We establish that the system of fuzzy ordered variational inclusions is equivalent to a fixed point problem and a relationship between a system of fuzzy ordered variational inclusions and a system of fuzzy ordered resolvent equations is shown. We prove the existence of a common solution and discuss the convergence of the sequence of iterates generated by the algorithm for a considered problem. The iterative algorithm and results demonstrated in this article have witnessed, a significant improvement for many previously known results of this domain. Some examples are constructed in support of the main results.

Hospital acquired-Staphylococcus aureus (HA-Staphylococcus aureus), particularly methicillin-resistant Staphylococcus aureus (MRSA), are an important source of nosocomial infections with high morbidity and mortality rates. Few reports showed that infections due to HA-Staphylococcus aureus in Saudi Arabia is increasing, particularly infections attributed to HA-MRSA. The study aimed to explore the prevalence and clinical characteristics of HA-Staphylococcus aureus for the first time in Medina, Saudi Arabia. A total of 1262 clinical samples of hospitalized patients were examined for the presence of Staphylococcus aureus through selective culturing on mannitol salt agar. Vitek Compact System and conventional methods were followed to confirm the isolates. Vitek Compact System tested the antimicrobial susceptibility of isolates whereas the standard PCR was employed to detect the genes encoding antimicrobial resistance (mecA and vanA) and virulence factors (tst, et, and LukS-PV). The overall HA-Staphylococcus aureus prevalence was low (6.58%, n = 1262) of which 84.34% (n = 83) were MRSA. Approximately, 57 samples of the 70 MRSA (81.5%) exhibited a multidrug-resistance (MDR) pattern. All the 83 HA-Staphylococcus aureus isolates were negative for the genes encoding toxic shock syndrome toxin, exfoliative toxin, and Panton-Valentine leukocidin. The study was conducted during the Covid-19 pandemic under partial lockdown, restricted hospitalization, and increased disinfection and infection control measures. Therefore, the low prevalence of HA-Staphylococcus aureus should be carefully interpreted and further multicenter investigations could reveal its true incidence in the city. The high prevalence of MDR HA-MRSA is alarming as it highlights inappropriate antibiotic prescriptions to counter staphylococcal infections. HA-Staphylococcus aureus investigated in this study might lack certain virulence factors. However, their MDR traits and invasive nature could worsen the situation if not properly handled.

Motivation plays a crucial role in determining students’ academic achievement. An important aspect of motivation is student achievement goals or Achievement Goal Orientation. The trichotomous model of Achievement Goal Orientation has been widely used: Mastery Goal Orientation, performance approach Goal Orientation, and Performance-Avoidance Goal Orientation. Educators must be familiar with the orientation students adopt, as it shapes their approach during teaching and learning. This paper examines the validity and reliability of Achievement Goal Orientation in a language education context. One hundred twenty-three secondary school students attended the survey, involving 55 (44.7%) males and 68 (55.3%) females. The instrument was adopted from Elliot and Church's Achievement Goal Orientation Questionnaire (1997) and was modified into the language domain. The initial version of the instrument consisted of 20 items, while the final version, after modification, consisted of 13 items. The Rasch Measurement Model was used to validate the instrument. All the psychometric properties confirmed the validity and reliability of the instrument. Rasch analysis revealed that the scale showed strong evidence of validity and reliability in the language education setting. It is suggested that language teachers and experts use the scale to improve language learners' performance. The findings of this study may be tested in other fields of study to get more robust validation results for further validation of the scale.

Emerging and re-emerging zoonoses of diverse etiologies have caused significant morbidity and mortality recently. In the past two decades, several viral zoonoses, such as Bird flu, Ebola hemorrhagic fever, Hantavirus infection, Nipah virus disease, Rift Valley fever, Swine flu, West Nile fever, SARS, MERS, COVID-19 etc., have emerged from different parts of the world. The latest to the list is the “Monkey Pox”, which has recently been renamed as “Mpox” by WHO. The ongoing 2022 multi-country outbreak of monkeypox is the largest in history to occur outside of Africa. Monkeypox is an emerging zoonotic disease that for decades has been viewed as an infectious disease with significant epidemic potential because of the increasing occurrence of human outbreaks in recent years. With increasing case numbers being reported in the current outbreak, it is important for healthcare staff everywhere to update their knowledge of this zoonotic infection, including its prevention, clinical management, prophylaxis, and basics of infection control, to understand the broader implications of the current outbreak. We provide an overview of monkeypox virus infection to serve as a primer for healthcare staff who may encounter this condition in their practice.

Exopolysaccharides (EPSs) are novel functional additives for low-fat yogurt. Pharmaceutical, medical, and food industries are using more LAB-based EPSs. In this study, Leuconostoc spp. was used to produce ninth bacterial EPSs in a modified molasses medium. Production of EPSs was concentration-dependent on all stains and the highest yield was obtained from the S3 strain (55.23 g/l), followed by S6 (49.95 g/l), S8 (45.68 g/l), and S7 (44.23), respectively. HPLC and FTIR analysis showed that all purified EPSs from Leuconostoc citreum (S3) and Leuconstoc holzaapfelii (S8) were related to exopolysaccharide glucan. Anticancer activity of all EPSs samples (EPSs1-9) against Caco-2 cells and normal MCR-5 cells were investigated using MTT assay. The results revealed that Caco-2 cells were more sensitive than the normal MCR-5 cells. The highest anticancer activity against Caco-2 cancer cells was recorded for EPS8 (IC50 = 22.94 µg/ml, SI=3.73), followed by EPS3 (IC50 = 36.15 µg/ml, SI=8.72), EPS1 (IC50 = 50.01 µg/ml, SI=3.73), and EPS4 (IC50 = 94.90 µg/ml, SI=3.26), respectively. The lowest cytotoxicity was recorded for EPS5 (IC50 = 130.5 µg/ml). The most active EPSs (EPS3 and EPS8) were used as fat replacements and stabilizers in low-fat set yogurt at non-toxic concentrations (0.4, 0.8, and 1.2%). EPS3 and EPS8 improved the low-fat yogurt’s organoleptic and rheological properties. EPS8 had the highest water holding capacity (77.26%), viscosity (3660 CP), and lowest syneresis (22.95%) and whey off (0.6 ml). Low-fat set yogurt enhanced with EPS3 and EPS8 recorded the highest sensory evaluation values with overall acceptability, especially EPS3b, EPS3c, EPS8c, and EPS8b; the total score point of 97.50, 97.43, 96.51, and 96.36, respectively in fresh age compared to control yogurt (92.64). In conclusion, Leuconostoc EPSs, especially EPS8, can be explored for anti-cancer effects on Caco-2 colorectal cancer cells. It could also improve the rheological and organoleptic qualities of low-fat set yogurt.

Proteus mirabilis is a specific opportunistic pathogen of many infections including urinary tract infections (UTIs). Risk factors are linked with the acquisition of multidrug-resistant (MDR) to 3 or more classes of antimicrobials) strains. The resistance in extended-spectrum alpha-lactamase is rare, but the rising resistance in extended-spectrum beta-lactamase (ESBL) producing strains is a matter of concern. β-lactamases and antibiotic modifying enzymes mainly constitute the ESBLs resistance mechanism by hydrolyzing the antibiotics. Mutation or Porin loss could lead to the reduced permeability of antibiotics, enhanced efflux pump activity hindering the antibiotic access to the target site, antibiotic failure to bind at the target site because of the target modification, and lipopolysaccharide mutation causing the resistance against polymyxin antibiotics. This review aimed to explore various antimicrobial resistance mechanisms in Proteus mirabilis and their impact on public health status.

This work is concentrated on the study of a system of mixed generalized Cayley variational inclusions. Parallel Mann iteration process is defined in order to achieve the solution. We define an altering point problem which is equivalent to our system and then we construct general parallel S-iteration process. Finally, we discuss convergence criteria and provide an example.

The selective degeneration of retinal ganglion cells (RGCs) is a common feature in glaucoma, a complex group of diseases, leading to irreversible vision loss. Stem cell-based glaucoma disease modeling, cell replacement, and axon regeneration are viable approaches to understand mechanisms underlying glaucomatous degeneration for neuroprotection, ex vivo stem cell therapy, and therapeutic regeneration. These approaches require direct and facile generation of human RGCs (hRGCs) from pluripotent stem cells. Here, we demonstrate a method for rapid generation of hRGCs from banked human pluripotent stem cell-derived retinal progenitor cells (hRPCs) by recapitulating the developmental mechanism. The resulting hRGCs are stable, functional, and transplantable and have the potential for target recognition, demonstrating their suitability for both ex vivo stem cell approaches to glaucomatous degeneration and disease modeling. Additionally, we demonstrate that hRGCs derived from banked hRPCs are capable of regenerating their axons through an evolutionarily conserved mechanism involving insulin-like growth factor 1 and the mTOR axis, demonstrating their potential to identify and characterize the underlying mechanism(s) that can be targeted for therapeutic regeneration.

SERINC5 is a host-restriction factor preventing HIV progeny entry, which is counteracted by interactions with HIV Nef. Here, Li et al. show that E3 ubiquitin ligase Cullin 3 polyubiquitinates SERINC5 at Lys 130 via K48- and K33-linked ubiquitin chains and provide evidence that this modification is not only required for its membrane localization and anti-viral activity but also relevant for Nef counteractive activity.

Biosynthesis of gold nanoparticles (AuNPs) is emerging as a better alternative to traditional chemical-based techniques. During this study, extracts of different marine algae species Ulva rigida (green algae), Cystoseira myrica (brown Algae), and Gracilaria foliifera (red Algae) were utilized as reducing and capping agents to synthesize AuNPs. AuNPs capped by U. rigida, C. myrica, and G. foliifera were confirmed by the appearance of surface plasmonic bands at 528, 540, and 543 nm, respectively. Transmission electron microscopy revealed mostly spherical shapes of AuNPs having a size of about 9 nm, 11 nm, and 13 nm for C. myrica, and G. foliifera extracts, respectively. Fourier transform-infrared spectroscopy (FTIR) illustrated the major chemical constituents of U. rigida, C. myrica, and G. foliifera. LC50 values of the biosynthesized AuNPs against Artemia salina nauplii were calculated at a range of concentrations (5-188 μg ml−1) after 16 to 24h. AuNPs concentration-dependent lethality was noted and U. rigida extracts-mediated AuNPs presented the lowest cytotoxicity. The biosynthesized AuNPs exhibited significant anticancer activity (86.83%) against MCF-7 cell lines (human breast adenocarcinoma cell lines) at 188 µg/ml concentration. G. foliifera demonstrated the highest anticancer value (92.13%) followed by C. myrica (89.82%), and U. rigida (86.83%), respectively. The AuNPs synthesized by different algal extracts showed variable antimicrobial activity against the tested pathogenic microorganisms. AuNPs of U. rigida extracts showed significant antimicrobial activity against dermatophytic fungi Trichosporon cataneum (30 mm) followed by Trichophyton mantigrophytes (25 mm). Furthermore, it also exhibited mild activity against Escherichia coli (17 mm), Cryptococcus neoformans (15 mm), Candida albicans (13 mm), and Staphylococcus aureus (11mm), respectively whereas no effects were observed against Bacillus cereus. To conclude, AuNPs can be effectively synthesized by marine algal species, and particularly U. rigida extracts could be effective reducing agents for the green AuNPs synthesis. These AuNPs could potentially serve as efficient alternative anticancer agents against human breast adenocarcinoma and anti-dermatophytes associated with skin infections.

BACKGROUND & OBJECTIVE: Thrombocytopenia is a common hematological manifestation of malaria, but locally there is limited data on the association of thrombocytopenia degree and malarial species in Pakistan. Therefore, the present study aims to assess the frequency of thrombocytopenia and its severity among patients with malaria. METHODOLOGY: In this prospective cross-sectional study, a total of 120 patients with smear-positive for malaria on microscopy were included. The patient's complete blood count (CBC) was assessed daily to monitor the platelet counts, and thrombocytopenia was divided into four categories, i.e., mild, moderate, severe, and very severe thrombocytopenia. RESULTS: Out of the total 120 malaria-positive cases, 78(65%) were positive for P. vivax, 32(26.7%) were affected by P. falciparum, and 10(8.3%) patients had mixed infection. Thrombocytopenia developed among 73% of the total patients with malaria; of these, 35.23% developed mild thrombocytopenia, while 31.82%, 23.86%, and 9.09% developed moderate, severe, and very severe thrombocytopenia, respectively. Among those 30(34.0%) infected with P. falciparum, 8(26.7%) developed mild thrombocytopenia, while 8(26.7%) and 14(46.7%) developed moderate to very severe thrombocytopenia. Similarly, most patients with P. vivax 23(39.7%) had mild to moderate 20(34.5%) thrombocytopenia, 15(25.9%) had severe, and only 5.19% had very severe thrombocytopenia p-value=0.141 shows non-significant association. Furthermore, P. vivax group developed complications, while two of the four patients of the P. falciparum group developed cerebral malaria, one developed renal failure, and the last one had pancytopenia. CONCLUSION: In conclusion, thrombocytopenia is frequent among patients with malaria. Moreover, the severity of thrombocytopenia and complications are more evident among those infected with P. falciparum than those with P. vivax.

In this article, our goal is to study the system of extended nonlinear mixed variational-like inequalities (in short, SENMVLI) with a nonconvex functional in the setting of real Hilbert spaces and discuss the existence of solution of our considered problem. We propose a three-step iterative algorithm to calculate the approximate solutions of SENMVLI and investigate the convergence analysis as well as stability analysis of the proposed algorithm. Furthermore, we also study the proximal dynamical system for SENMVLI and prove that the trajectory of the solution of the extended proximal dynamical system converges globally exponentially to a unique solution of SENMVLI. Our suggested iterative algorithm and results have become the significant improvement, enhancement, and generalization of many previously known results in the literature.

ABSTRACT: Imprudent and overuse of clinically relevant antibiotics in agriculture, veterinary and medical sectors contribute to the global epidemic increase in antimicrobial resistance (AMR). There is a growing concern among researchers and stakeholders that the environment acts as an AMR reservoir and plays a key role in the dissemination of antimicrobial resistance genes (ARGs). Various drivers are contributing factors to the spread of antibiotic-resistant bacteria and their ARGs either directly through antimicrobial drug use in health care, agriculture/livestock and the environment or antibiotic residues released from various domestic settings. Resistant micro-organisms and their resistance genes enter the soil, air, water and sediments through various routes or hotspots such as hospital wastewater, agricultural waste or wastewater treatment plants. Global mitigation strategies primarily involve the identification of high-risk environments that are responsible for the evolution and spread of resistance. Subsequently, AMR transmission is affected by the standards of infection control, sanitation, access to clean water, access to assured quality antimicrobials and diagnostics, travel and migration. This review provides a brief description of AMR as a global concern and the possible contribution of different environmental drivers to the transmission of antibiotic-resistant bacteria or ARGs through various mechanisms. We also aim to highlight the key knowledge gaps that hinder environmental regulators and mitigation strategies in delivering environmental protection against AMR.

Multidrug resistance (MDR) in pathogenic bacteria have become a major clinical issue. Quorum sensing regulated bacterial virulence is a promising key drug target for MDR infections. Therefore, the aim of the present work was to assess the anti-quorum sensing properties of selected medicinal plants against bacterial pathogens as well in silico interaction of selected bioactive phytocompounds with QS and biofilm-associated proteins. Based on the ethnopharmacological usage, 18 plants were selected using methanolic extract against Chromobacterium violaceum 12472. The most active extract (Acacia nilotica) was fractionated in increasing polarity solvents (n-hexane, chloroform and ethyl acetate) and tested for anti-QS activity. The most active fraction i.e. ethyl acetate fraction was evaluated for their activity at sub-MICs against QS-associated virulence factors of Pseudomonas aeruginosa PAO1 and Serretia marcescens MTCC 97. Microtiter plate assay and light microscopy was used to determine inhibition of biofilm. Phytochemicals of the ethyl acetate fraction were analysed by GC/MS and LC/MS. Phytocompounds were docked with QS (LasI, LasR, CviR, and rhlR) and biofilm proteins (PilY1 and PilT) using Auto dock vina. The MIC of ethyl acetate fraction determined was 250, 500, and 1000 μg/ml against C. violaceum 12472, P. aeruginosa PAO1, and S. marcescens MTCC97 respectively. At sub-MICs QS regulated virulence factors production and inhibited biofilms broadly (more than 50 percent). GC/MS detected the major bioactive compound benzoic acid, 3,4,5-trihydroxy-, methyl ester (61.24 %) and LC-MS detected Retronecine for the first time in A. nilotica pods. In silico, dehydroabietic acid occupied the same cavity as its antagonist in the CviR ligand binding domain. Also, betulin and epicatechin gallate interact with biofilm proteins PilY1 and PilT, preventing biofilm formation. The findings suggest that the phytochemicals of A. nilotica pod could be exploited as an anti-QS agent against Gram-negative pathogens. To discover therapeutic efficacy of standardised bioactive extract/phytochemicals must be tested under in vivo condition.

An effort has been made to synthesize nanocomposites of TiO2/jackfruit peel via green synthesis. Various concentration of the synthesized nanocomposite (0.1%, 0.2% and 0.3%) has been combined with mat black paint to coat the sides and base of the proposed single slope single basin solar still (PSBSS). The structural and morphological properties of synthesized nanocomposite (JPT) were characterized using XRD and SEM. It was observed that the nanocomposite of JPT has a porous structure with 85% crystallinity with grain size ranging from 40 to 60 nm. The nanocomposite is mixed with cobalt (II) chloride (CoCl2), thiourea (CH4N2S) and silicon dioxide (SiO2) to make a new hybrid nanofluid (CTS) with a ratio of 10%, 20%, 30%, 40%, 50% and 60%. Silver-color steel balls filled with CTS was placed in the basin of the PSBSS at equal intervals to enhance the heat transfer process inside the still. The PSBSS with nanocomposites (0.3%), nanofluids (40%) fill the silver-color steel balls (JPTCTSS) has high productivity of 8.7919 L/m2day, which is 50.55% higher than that of the conventional solar still (CSS) with nanofluid along with the saline water in the basin.

In submitting the proceedings of the 5th Conference on Sustainability in Civil Engineering (CSCE) to Engineering Proceedings, the volume editors of the proceedings certify to the publisher that all papers published in this volume have been subjected to peer review administered by the volume editors [...]

The Conference on Sustainability in Civil Engineering (CSCE), 2023, was organized by the Department of Civil Engineering of the Capital University of Science and Technology (CUST), Islamabad, Pakistan [...]

Abstract Purpose Rhizospheric soil fungi are critical for plant and soil health. However, their multiple functional traits and impact on plant growth have not been systematically explored. Methods During this study, biochemical traits of 73 indigenous soil fungal isolates and 15 unidentified isolates related to plant growth promotion and production of extracellular enzymes were studied. Results Forty four (65.67%) of the total isolates produced indole acetic acid (IAA) followed by siderophore (52.23%), phosphate solubilization (37.31%), and antibiotic (11.93%). 91.04% of the studied isolates produced ammonia whereas 28.35% produced organic acid. Extracellular enzyme activities of lipase, amylase, chitinase, and cellulase were detected among 95.52%, 61.11%, 35.82%, and 41.79% isolates, respectively. Based on these activities, 73 fungal isolates were categorized into different biotypes. Quantitative analysis of IAA production and phosphate solubilization was carried out for Aspergillus, Penicillium, and Rhizopus isolates. Aspergillus isolates exhibited varying activities of IAA production and phosphate solubilization. Most of the Aspergillus isolates and some other fungi demonstrated multiple activities. Based on the multiple traits of selected fungal isolates, Aspergillus sp-07, Penicillium sp-03, and Rhizopus sp-02 were further evaluated in different combinations for their inoculation effect on the growth and yield of wheat under field conditions. Conclusions The results indicated that these isolates could be developed into bio-inoculants to enhance plant growth. The consortium of these three isolates was also found to be compatible and beneficial for plant growth.

Plastic materials have become a necessity of human life especially in the packaging of food commodities and biomedical procedures. Bioplastic is emerging as an effective alternative to fossil oil-based materials to avoid the environmental hazards of the plastic industry. During this study, chicken feathers were used as a substrate to isolate keratin degrading bacteria. Among 14 identified isolates, Bacillus sp BAM3 was found to be the most promising isolate. Partial 16S rDNA analysis-based molecular characterization revealed it is a strain of Bacillus cereus. Bacillus sp BAM3 can grow and produce keratinase in feathers containing basal medium as the sole carbon and energy source. The maximum keratinase production (730U/ml) was achieved within 24 h under optimum reaction conditions. The optimized reaction pH and temperature were noted as 9.0 and 50 °C for crude keratinase activity, respectively. The chicken feathers were used as a substrate in 2, 5, and 10 wt% glycerol to synthesize keratin-based bioplastic with keratinolytic bacterium Bacillus cereus BAM3. Bioplastic prepared from keratin with 2% of glycerol was found to possess good mechanical properties. Therefore, the results present a novel keratinolytic isolate of Bacillus cereus BAM3, which may have potential biotechnological applications in keratin hydrolysis processes. The development of keratin-based bioplastics possessing superior crystalline morphology requires further investigations to substitute fossil oil-based materials.

We report in this study for the first time the prevalence of multiple resistant Staphylococcus haemolyticus in clinical settings in Saudi Arabia. A total of 1060 clinical specimens of hospitalized patients were screened for the presence of S. haemolyticus in the period between September and December 2020. Primary identification of the isolates was carried out by colonial characteristics on mannitol salt agar and clumping factor test, confirmation of presumptive isolates and antimicrobial susceptibility testing was performed by Vitek® 2, while PCR was employed to detect mecA and vanA genes. A total of 20 S. haemolyticus isolates were recovered from 20 samples (blood cultures, urine, nasal swab, wound swab, groin swab, and axilla swab), 90% (P

Staphylococcus aureus is a major human pathogen that may cause a wide range of infections and is a frequent cause of soft tissue and bloodstream infections. It is a successful pathogen due to its collective virulence factors and its ability to evade the host immune systems. The review aims to highlight how S. aureus destroys and damage the host cells and explains how immune cells can respond to this pathogen. This review may also provide new insights that may be useful for developing new strategy for combating MRSA and its emerging clones such as community-associated methicillin-resistant S. aureus (CA-MRSA).

Objective: To determine patient’s level of satisfaction and factors leading to dissatisfaction in selected patients group with health care delivery in outpatients department of CMH Rawalpindi. Study Design: Cross sectional study. Place and Duration of Study: Out Patients Department of Combined Military Hospital Rawalpindi, from Sep 2013 to Aug 2015. Methodology: Before data collection, written informed consent was taken from all the participants. The study population comprised of armed forces personnel and their families. Patients, including both, males and females, coming to the outpatient department of Combined Military Hospital Rawalpindi were included in the study through consecutive sampling. Results: Results revealed that only 30% were satisfied. 67% patients were satisfied with the facilities in reception and 70% were satisfied with cleanliness in waiting area. A relatively low level of satisfaction with dealing of nursing assistant was recorded while very high percentage of satisfaction with performance of doctor i.e. 91%, examination done by doctor 91% and information provided by doctor 93% was recorded.90% patients reported high level of satisfaction with medicines provided.95.33% patients were satisfied with lab tests being done in hospital. Conclusion: Assessing satisfaction of patients is a simple and cost effective way for assessment of hospital services. Most of the patients were contented with services delivered in OPD and showed their trust in doctors, medicines provided and lab facilities offered. Majority were willing to revisit the hospital if required.

Abstract In vitro eradication of the C. albicans and S. mutans mixed biofilms by eugenol alone and in combination with the antimicrobial drugs. Previously characterized strains of C. albicans (CAJ-01 and CAJ-12) and S. mutans MTCC497 were used to evaluate the eradication of biofilms using XTT reduction assay, viability assay, time dependent killing assay and scanning electron microscopy (SEM). Synergistic interaction was assessed by checkerboard method. Sessile MIC (SMIC) of eugenol was equivalent to the planktonic MIC (PMIC) against C. albicans and S. mutans mixed biofilms. SMIC of fluconazole and azithromycin was increased upto 1000-folds over PMIC. Eradication of single or mixed biofilms was evident from the viability assay and SEM. At 1 × MIC of eugenol, log10CFU count of C. albicans cells were decreased from 6.3 to 4.2 and 3.8 (p

Developing cost-effective and more efficient nanocatalysts for the treatment of organic pollutants from process industry is always challenging for the researchers working in the field of chemistry, chemical, energy and environment engineering. In this work, a cost-effective and more efficient nanocatalysts, i.e., Nickel Cobaltite nanocomposites and its Lanthanum (La) doped derivatives with controlled surface morphology has been synthesized at 393.15 K through single step sol–gel method. The surface morphology, chemical composition, and crystal structure of the synthesized nanocomposites were analysed by scanning electron microscopy (SEM), Fourier transformed infrared spectroscopy (FTIR), and X-rays diffraction (XRD), respectively. The rough surface and well-crystallized metallic nanocomposites confirm the successful synthesis of nanocatalysts. The molar ratio of Lanthanum to Cobalt (Lax:Coy) showed a significant influence on the surface morphology and catalytic activity (Kapp = 0.15–0.47 min−1) of the products. Synthesized nanocomposites showed high catalytic activity for the reduction of methylene blue under solar irradiation. Photocatalytic results for the reduction of methylene blue show that the catalytic activity of synthesized nanocatalysts increases with the increase in the doping concentration of Lanthanum.

In this study fungal profiles of agricultural field soil irrigated with industrial wastewater and sewage containing varying concentrations of heavy metals (Chromium, Nickel, Cobalt, Copper and Cadmium) have been investigated. The impact of long term heavy metal contamination on emergence of heavy metal tolerant soil fungal population, changes in morphological diversity and metal tolerance limits among isolated fungi was studied. The agricultural field soil received long term (>20 years) wastewater application showed metal accumulation compared to the untreated soil. The viable count of soil fungal population from three different agricultural field soil was found in order of 105 to 10 4 CFU gm-1 of soil indicating a normal viable count with little variations. Viable plate count of fungal population on metal amended plates decreased with increasing concentration of tested metals (Cr6+, Cd++. Cu++, Co++ and Ni++) from 100 to 400 µgml-1. The decrease was higher on cadmium amended plates and lower against Chromium. The control site, which did not receive wastewater application showed relatively less metal tolerant fungal viable count on Cd++ and Ni++ plates when compared at 100 µgml-1as compared to contaminated sites. Similarly, presence of metal tolerant fungal population was also observed from wastewater sample. The common soil fungi isolated and characterized from metal amended plates belong to 18 genera and 15 unidentified species. Occurrence of different fungal genera from site A B and C indicated different patterns of decrease on different metal amended plates with increasing concentration. Among these 73 isolated fungal species a high level of tolerance was recorded to Cr6+ followed by Cu++, Co++ and Ni++ while the lowest level of tolerance was for Cd++. The minimum inhibitory concentration (MIC) values of 73 metal tolerant fungal isolates, was ranged from 200 to 2000 μgml-1 against one or more heavy metals. The level of tolerance to heavy metals also varied even among the isolates of single genus. Aspergillus was the predominant genus recovered from contaminated soils where the MIC values are highly varied among different isolates of Aspergillus. The current study found that long term release of wastewater has not disturbed the fungal population dynamics in contaminated sites as compared to uncontaminated sites. However, it tends to exert selective pressure on fungal populations of soil, leading to the development of increased level of metal tolerance in fungal species.

Abstract In the setting of real ordered positive Hilbert spaces, a nonlinear fuzzy ordered variational inclusion problem with its corresponding nonlinear fuzzy ordered resolvent equation problem involving XOR operation has been recommended and solved by employing an iterative algorithm. We establish the equivalence between nonlinear fuzzy ordered variational inclusion problem and nonlinear fuzzy ordered resolvent equation problem. The existence and convergence analysis of the solution of nonlinear fuzzy ordered variational inclusion problem involving XOR operation has been substantiated by applying a new resolvent operator method with XOR operation technique. The iterative algorithm and results demonstrated in this article have witnessed a significant improvement in many previously known results of this domain.

Context: Directly observed treatment short course (DOTS) was adopted as the strategy for the provision of treatment to increase treatment completion. Poor adherence to medication has been noted and poses a big challenge even after achieving 70% case detection and 85% cure rate. The focus remains on dealing with important reasons of default and timely retrieval of patients who interrupt the treatment. Aims: The aim of the study was to measure the noncompliance of DOTS and to determine the reason for the same among patients in rural area of Pune. Materials and Methods: An observational cross-sectional study was conducted with a study population comprising the patients who were getting treatment in November 2016 and must have completed at least 1 month of treatment in Mulshi block. The study period was from November 2016 to January 2017. All the patients were interviewed using a pretested questionnaire for their compliance. Results: The total number of patients included in the study was 88. The age was 18–70 years in which 77.3% were male and 22.7% were female. We found 71.6% new cases, and 27.3% were the previously treated patients, whereas 1.1% were multidrug-resistant patients. Among these patients, 25% were tobacco chewers and 31.8% were smokers. We found that 61.4% were compliance patients and 38.6% were noncompliance patients. Reasons given by the patients were side effects, stigma of the society, migration from one place to another, and felt better from symptoms after taking medication. Conclusion: Noncompliance in our study is high. We should take steps regarding this so that people adhere to the medication.

Ever increasing spread of drug resistance among bacterial pathogens have rendered the current antibiotic therapy ineffective. Drug-resistant urinary tract infections caused by Gram negative bacterial pathogens such as Pseudomonas aeruginosa, Escherichia coli, Acinetobacter baumannii, and Serratia marcescens are considered more severe and life threatening because of strong biofilm dependent growth characteristics among majority of them. Moreover, most uropathogens coordinate their complex virulence responses via a highly structured network of cell to cell communication known as Quorum sensing (QS). Phytochemicals/bioactive extracts obtained from established ethnomedicinal plants have previously been demonstrated as effective anti-QS agent against numerous pathogenic bacteria including uropathogens. Myrtus communis (L.) (Myrtaceae) is one such plant of immense ethnomedicinal importance and has been used in traditional medicine. In the present investigation, methanolic extract of Myrtus communis (MCME) demonstrated 65% inhibition in the QS regulated violacein production in C. violaceum without affecting the growth of the bacteria. Further, the MCME interfered significantly with QS controlled virulence production in P. aeruginosa (elastase, protease, pyocyanin and chitinase) and S. marcescens (prodigiosin and protease). The leaf extract exhibited 16–74%, 31–84%, 12–66% and 19–71% inhibition of biofilm biomass of P. aeruginosa, E. coli, A. baumannii and S. marcescens respectively at the increasing concentration corresponding to MIC/16-MIC/2. GC–MS analysis revealed Linalool as one of the major bioactive compounds present in the MCME. Further, in vitro assays demonstrated QS and biofilm inhibition by sub-MICs of linalool. In silico docking and simulation studies showed that the possible mechanisms responsible for the interference of QS were AHL synthesis inhibition, antagonization of QS-regulatory proteins, and blocking of the receptor proteins. Thus, from the obtained results it is envisaged that M. communis and its bioactive compound linalool may have medicinal implications and could prove as effective therapeutic agent in combating the threat of biofilm based persistent infections caused by uropathogenic bacteria.

Summary: HIV-1-negative factor (Nef) protein antagonizes serine incorporator 5 (SERINC5) by redirecting this potent restriction factor to the endosomes and lysosomes for degradation. However, the precise mechanism remains unclear. Using affinity purification/mass spectrometry, we identify cyclin K (CycK) and cyclin-dependent kinase 13 (CDK13) as a Nef-associated kinase complex. CycK/CDK13 phosphorylates the serine at position 360 (S360) in SERINC5, which is required for Nef downregulation of SERINC5 from the cell surface and its counteractivity of the SERINC5 antiviral activity. To understand the role of S360 phosphorylation, we generate chimeric proteins between CD8 and SERINC5 to study their response to Nef. Nef not only downregulates but, importantly, also binds to this chimera in an S360-dependent manner. Thus, S360 phosphorylation increases interactions between Nef and SERINC5 and initiates the destruction of SERINC5 by the endocytic machinery.

Effective rhizosphere colonization by plant growth promoting rhizobacteria (PGPR) is a prerequisite for its persistence performance under plant-root soil system. Therefore, to obtain effective strains, we have evaluated twelve strains of fluorescent Pseudomonas sp. from wheat rhizosphere exhibiting multifarious plant growth promoting (PGP) activities. These strains formed varying level of biofilm in vitro on abiotic surface. Two strains MW3 and MW4 with promising PGP activity but differing in biofilm forming ability. The biofilms were characterized by crystal violet assay, scanning electron and confocal laser scanning microscopy. The MW4 strain, a strong biofilm former is identified by16S rRNA gene sequence analysis. Strain MW4 showed strong biofilms compared to MW3. Similarly, wheat seeds treated with MW3 and MW4 revealed enhanced rhizospheric colonization by MW4 strain significantly (P ≤ 0.05) in higher number (logCFU g-1) up to 45 days after sowing compared to MW3. These findings suggested that strong biofilm forming ability of MW4 might have resulted in enhanced rhizosphere colonization. Therefore, biofilm forming ability of PGPR should be considered as an additional criterion in screening of fluorescent Pseudomonas sp.

Exopolysaccharides (EPS) particularly, from Lactic acid bacteria have received increasing attention in food, medical, and pharmaceutical applications. The present work aims to isolate, characterize and identify exopoly saccharide-producing bacteria from fermented fruits and vegetables and dairy products. A total of 55 isolates were isolated from fermented fruits, vegetables, and dairy products depending on the mucoid appearance of the colonies. Based on total EPS production, the most promising nine strains were selected, phenotypically and genotypically characterized. They were facultative anaerobe, arranged in pairs/chains (cocco bacillus), oxidase, and catalase-negative, non-spore forming and non-motile Gram-positive bacteria. All the strains were capable of growing at optimum pH between 5-7, tolerate to NaCl up to 7% (w/v), growing at 20-37°C with optimum growth at 30°C, no growth was observed at 45°C. In addition they could utilize small range of organic compounds, except isolate S1 was differ from the others by their ability to utilize a varied range of organic compounds. Construction of phylogenetic tree, on the basis of partial 16S rRNA gene sequences indicated that isolate S1 was similar to Leuconostoc citreum with similarity of 91.3%, while, isolates S2 and S3 were similar to Leu. fallax and Leu. mesenteroides with similarity of 99.40 % and 97.73%, respectively. Isolates S4, S5, S7, S8, and S9 were similar to Leu. holzaapfelii with similarity of 98.3, 98.7 and 99.8, 98.5 and 98.1, respectively, while isolate S6 was similar to Leu. lactis with similarity of 97.9%. None of sugars such as lactose, glucose, and fructose except sucrose were support EPS production from these strains. The highest yield of EPS was recorded for isolates S6, S1 and S7 which were 61.90, 61.80 and 60 gl-1, respectively, followed by isolates S4, S9, S5 and S8 which were 58.40, 53.06, 51.61 and 33.53 gl-1, respectively. Although, the lowest yield was observed for the isolates S3 and S2 which were 22.08 and 18.80 g l-1, respectively. Finally, it could be concluded that EPS production from these strains in the current study, considering them to be the alternative choice for enhancing production of EPS with increased yields, with promising realistic importance in food, pharmaceutical, as well as dairy industries.

Heavy metal contamination due to natural and anthropogenic source is a major environmental problem. Release of metal from use of agrochemicals, industrial effluents and wastewater residues and their accumulation in food causes serious dilemma to animal and human health concern. On the other hand microbial population gets affected from metal toxicity at elevated concentration. With the result microbes develops various resistance mechanism to cope with metal toxicity. Both physiological and genetic mechanisms are involved in developing such resistance. Recent advances on metal-bacteria interaction has led to better understanding of metal accumulation/detoxification or biotransformation and bioremediation of metals through application of such bacteria. Role of various transport protein families involved in heavy metal metabolism are now explored. This article provides insights of metal-bacteria interaction in terms of resistance mechanisms and role of various transport proteins and its potential application in bioremediation of metal pollution.

Background: Malnutrition continues to be a major public health problem in developing and underdeveloped countries. The objectives of the study were to determine the determinants of nutritional status of under 5 years children in our population. Materials & Methods: This analytical cross sectional study was conducted by the Department of Public Health, Islamabad, Federal College, Islamabad, from November 2016 to March 2017. Sample size was 200 selected by convenient sampling. Demographic variables were sex, age-groups, socio-economics status, family structure. Research variable was nutritional status. The data collection was done in urban slums of Islamabad on a structured questionnaire. Statistical analysis was done by using SPSS software. Results: Out of 200 children, 117(58.5%) were boys and 83(41.5 %) were girls. One hundred (50%) children were of 6 to 24 months and 100(50%) were of 25 to 59 months of age. Eighty three (41.5%) were rich and 117(58.5%) were poor. One hundred and fifty eight (79%) children were from joint family system and 42(21%) were living in nuclear family system were children. One hundred and fifty one (75.50%) were normal, 39(19.50%) were mild and 10(5%) were having severe malnutrition. Conclusion: Except age all the other factors including sex, socio-economics status and family structure were associated with nutritional status of under 5 year children.

In this study, we introduce and study a generalized complementarity problem involving XOR operation and three classes of generalized variational inequalities involving XOR operation. Under certain appropriate conditions, we establish equivalence between them. An iterative algorithm is defined for solving one of the three generalized variational inequalities involving XOR operation. Finally, an existence and convergence result is proved, supported by an example.

In this paper, we study a set-valued extended generalized nonlinear mixed variational inequality problem and its generalized resolvent dynamical system. A three-step iterative algorithm is constructed for solving set-valued extended generalized nonlinear variational inequality problem. Convergence and stability analysis are also discussed. We have shown the globally exponential convergence of generalized resolvent dynamical system to a unique solution of set-valued extended generalized nonlinear mixed variational inequality problem. In support of our main result, we provide a numerical example with convergence graphs and computation tables. For illustration, a comparison of our three-step iterative algorithm with Ishikawa-type algorithm and Mann-type algorithm is shown.

ABSTRACT Membrane-associated RING-CH-type 8 (MARCH8) strongly blocks human immunodeficiency virus type 1 (HIV-1) envelope glycoprotein (Env) incorporation into virions by downregulating its cell surface expression, but the mechanism is still unclear. We now report that MARCH8 also blocks the Ebola virus (EBOV) glycoprotein (GP) incorporation via surface downregulation. To understand how these viral fusion proteins are downregulated, we investigated the effects of MARCH8 on EBOV GP maturation and externalization via the conventional secretion pathway. MARCH8 interacted with EBOV GP and furin when detected by immunoprecipitation and retained the GP/furin complex in the Golgi when their location was tracked by a bimolecular fluorescence complementation (BiFC) assay. MARCH8 did not reduce the GP expression or affect the GP modification by high-mannose N-glycans in the endoplasmic reticulum (ER), but it inhibited the formation of complex N-glycans on the GP in the Golgi. Additionally, the GP O-glycosylation and furin-mediated proteolytic cleavage were also inhibited. Moreover, we identified a novel furin cleavage site on EBOV GP and found that only those fully glycosylated GPs were processed by furin and incorporated into virions. Furthermore, the GP shedding and secretion were all blocked by MARCH8. MARCH8 also blocked the furin-mediated cleavage of HIV-1 Env (gp160) and the highly pathogenic avian influenza virus H5N1 hemagglutinin (HA). We conclude that MARCH8 has a very broad antiviral activity by prohibiting different viral fusion proteins from glycosylation and proteolytic cleavage in the Golgi, which inhibits their transport from the Golgi to the plasma membrane and incorporation into virions. IMPORTANCE Enveloped viruses express three classes of fusion proteins that are required for their entry into host cells via mediating virus and cell membrane fusion. Class I fusion proteins are produced from influenza viruses, retroviruses, Ebola viruses, and coronaviruses. They are first synthesized as a type I transmembrane polypeptide precursor that is subsequently glycosylated and oligomerized. Most of these precursors are cleaved en route to the plasma membrane by a cellular protease furin in the late secretory pathway, generating the trimeric N-terminal receptor-binding and C-terminal fusion subunits. Here, we show that a cellular protein, MARCH8, specifically inhibits the furin-mediated cleavage of EBOV GP, HIV-1 Env, and H5N1 HA. Further analyses uncovered that MARCH8 blocked the EBOV GP glycosylation in the Golgi and inhibited its transport from the Golgi to the plasma membrane. Thus, MARCH8 has a very broad antiviral activity by specifically inactivating different viral fusion proteins.