Acyclovir-Induced Nephrotoxicity in Adults with Viral Encephalitis.

OBJECTIVE: To determine the frequency of parenteral Acyclovir-induced Acute Kidney Injury (AKI) in patients with viral encephalitis. STUDY DESIGN: Descriptive study. Place and Duration of the Study: Department of Neurology, Liaquat National Hospital, Karachi, from January to December 2021. METHODOLOGY: A total of 89 suspected and proven cases of encephalitis receiving IV Acyclovir were collated. All had extensive medical histories and underwent CSF studies with +/- brain imaging. CSF routine and viral PCR were done. Acyclovir-induced AKI was defined as a rise in serum creatinine of >0.3 mg/dl in 48 h or by ≥1.5 times the baseline value, and its severity was staged into 1 (risk), 2 (injury), and 3 (failure) according to the KDIGO guidelines (Kidney Disease: Improving Global Outcomes (KDIGO) Acute Kidney Injury Work Group, 2012). Patients' variables, including age, gender, presenting features, comorbid conditions, and CSF findings, were divided into two groups, i.e. with and without AKI. RESULTS: This research included 89 patients with a mean age of 48 years. AKI occurred in 34 patients (38.2%). The frequency of AKI with Stage 1 was 24%, Stage 2 was 44%, and Stage 3 was 32%; approximately two-thirds of cases were in Stage 2 and 3 (p >0.05). Five patients (5.6%) from Stage 3, required dialysis. CONCLUSION: AKI is an important adverse effect of parenteral acyclovir, which necessitates its early identification and timely management. Renal function monitoring is essential for patients on Acyclovir treatment as they are at risk for AKI. KEY WORDS: Acyclovir, Acute kidney injury, Viral encephalitis, Creatinine, Kidney Disease Improving Global Outcomes.

Chromate Removal by Enterobacter cloacae Strain UT25 from Tannery Effluent and Its Potential Role in Cr (VI) Remediation.

An indigenous chromate-resistant bacterial strain isolated from tannery effluent was identified based on morphological, biochemical, and 16S rRNA gene sequencing, as Enterobacter cloacae UT25. It was found to resist heavy metal ions such as Cr (VI), Pb (II), Cu (II), Co (II), Ni (II), Hg (II), and Zn (II) and antibiotics. The strain was able to remove 89 and 86% chromate, after 24 h of incubation in a Luria-Bertani (LB) medium at an initial Cr (VI) concentration of 1000 and 1500 µg/ml, respectively. Minimum inhibitory concentration (MIC) was observed for chromate to be 80,000 and 1850 µg/ml, after 48 h of incubation in LB and acetate minimal media (AMM), respectively. Scanning Electron Microscopy (SEM) and Transmission Electron Microscopy (TEM) analysis showed discrete cells with intact and smooth cell walls and homogenous cytoplasm in the absence of metal stress, whereas chromate stress caused cell lysis and reduction in size, which was a characteristic response to Cr (VI) toxicity. Energy Dispersive X-Ray Spectroscopy (EDX) confirmed the adsorption of oxyanions to the cell wall which was one of the Cr (VI) removal mechanisms by the bacterium. Atomic Force Microscopy (AFM) micrographs of chromate-untreated and treated cells revealed Root Mean Square roughness (Rq) values of 16.25 and 11.26 nm, respectively, indicating less roughness in the presence of stress. The partial gene sequence of class 1 integrons (intI1) of strain UT25 showed 94% homology with intI1 gene of strain Enterobacter hormaechei strain ECC59 plasmid pECC59-1. The present analysis highlighted the potential of E. cloacae UT25 as a promissory bacterium that could be applied in removing chromate from polluted environments.

Co-Culture of Halotolerant Bacteria to Produce Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) Using Sewage Wastewater Substrate.

The focus of the current study was the use of sewage wastewater to obtain PHA from a co-culture to produce a sustainable polymer. Two halotolerant bacteria, Bacillus halotolerans 14SM (MZ801771) and Bacillus aryabhattai WK31 (MT453992), were grown in a consortium to produce PHA. Sewage wastewater (SWW) was used to produce PHA, and glucose was used as a reference substrate to compare the growth and PHA production parameters. Both bacterial strains produced PHA in monoculture, but a copolymer was obtained when the co-cultures were used. The co-culture accumulated a maximum of 54% after 24 h of incubation in 10% SWW. The intracellular granules indicated the presence of nucleation sites for granule initiation. The average granule size was recorded to be 231 nm; micrographs also indicated the presence of extracellular polymers and granule-associated proteins. Fourier transform infrared spectroscopy (FTIR) analysis of the polymer produced by the consortium showed a significant peak at 1731 cm-1, representing the C=O group. FTIR also presented peaks in the region of 2800 cm-1 to 2900 cm-1, indicating C-C stretching. Proton nuclear magnetic resonance (1HNMR) of the pure polymer indicated chemical shifts resulting from the proton of hydroxy valerate and hydroxybutyrate, confirming the production of poly(3-hydroxybutyrate-co-3-hydroxy valerate) (P3HBV). A 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay showed that the copolymer was biocompatible, even at a high concentration of 5000 µg mL-1. The results of this study show that bacterial strains WK31 and 14SM can be used to synthesize a copolymer of butyrate and valerate using the volatile fatty acids present in the SWW, such as propionic acid or pentanoic acid. P3HBV can also be used to provide an extracellular matrix for cell-line growth without causing any cytotoxic effects.

Evaluating the Role of Wastewaters as Reservoirs of Antibiotic-Resistant ESKAPEE Bacteria Using Phenotypic and Molecular Methods.

Introduction: Wastewaters carrying thousands of human specimens from the community and representing the diversity of antibiotic-resistant bacteria (ARB) and antibiotic-resistance genes (ARGs) directly from the community mirror the extent of AR spread in the community and environment. This study aimed to investigate the occurrence and distribution of antibiotic-resistant ESKAPEE bacteria in the community versus clinical settings through monitoring nonclinical and clinical wastewaters. Methodology: Seven wastewater samples were collected from different environmental sources. Isolates were obtained on general and selective media, biochemically characterized and antimicrobial-susceptibility tests performed by disk diffusion against 13 antibiotics according to the European Committee on Antimicrobial Susceptibility Testing (EUCAST) guidelines using MastDisc disk cartridges, and 16S rRNA metagenomic analysis was performed for two water samples. Results: Of 43 isolates, all representatives of the ESKAPEE group were recovered from clinical wastewaters, but Gram-positive cocci were not obtained from nonclinical wastewaters. The most predominant isolate was Pseudomonas aeruginosa (n=15; 33%), followed by Escherichia coli (n=9; 20%). Complete (100%) resistance to eleven of the tested antibiotics was observed, with only a few isolates being susceptible to clarithromycin, amikacin, and gentamicin. The lowest (79%) resistance rate was observed for linezolid. The multiple antibiotic resistance (MAR) index was calculated, and the resistance phenotype was independent of the wastewater source, indicated by x 2 (P=0.766). Metagenomic analysis replicated the results, as Pseudomonas spp., Acinetobacter spp., and Escherichia spp. were found to be predominant. The integrase gene (IntI1) was also amplified in Escherichia coli, Klebsiella pneumoniae, and Pseudomonas aeruginosa. Conclusion: Wastewaters are significant carriers of drug-resistant ESKAPEE bacteria and play an important role in their dissemination. This study endorses the periodic surveillance of water systems to evaluate the presence and burden of antibiotic-resistant pathogens.

Seasonal variation and spatial distribution of microplastics in tertiary wastewater treatment plant in South Korea.

Wastewater treatment plant (WWTP) is proposed to be a point source of microplastics (MPs) that enter aquatic environments. Here, we quantified and characterized MPs down to 20 µm at a tertiary WWTP in Korea over a 1-year period. All wastewater contained MPs during the monitoring period, with concentrations ranging from 114 ± 17-216 ± 65 particles/L for influent and from 0.26 ± 0.29-0.48 ± 0.11 particles/L for effluent. MP abundance in the influents showed significant seasonal differences whereas a seasonal trend was not observed for the effluents, indicating a stable treatment performance (approx. 99.8%) of the WWTP. Spatial surveys of MP distribution in the WWTP showed that most MPs were removed by coagulation-sedimentation and rapid sand filtration, but some MPs still remained in the final effluent to generate the potential annual load of 2.9 × 109 particles and 0.54 kg into the rivers. These findings highlight the importance of long-term monitoring and MP mass quantification that would promote more accurate estimation of the MP load from WWTPs.

Biodeterioration of Microplastics: A Promising Step towards Plastics Waste Management.

Polyethylene and Polyester materials are resistant to degradation and a significant source of microplastics pollution, which is an emerging concern. In the present study, the potential of a dumped site bacterial community was evaluated. After primary screening, it was observed that 68.5% were linear low-density polyethylene, 33.3% were high-density, and 12.9% were Polyester degraders. Five strains were chosen for secondary screening, in which they were monitored by FTIR, SEM and weight loss degradation trials. Major results were observed for Alcaligenes faecalis (MK517568) and Bacillus cereus (MK517567), as they showed the highest degradation activity. Alcaligenes faecalis (MK517568) degrades LLDPE by 3.5%, HDPE by 5.8% and Polyester by 17.3%. Bacillus cereus (MK517567) is better tolerated at 30 °C and degrades Polyester by 29%. Changes in infrared spectra indicated degradation pathways of different strains depending on the types of plastics targeted. Through SEM analysis, groves, piths and holes were observed on the surface. These findings suggest that soil bacteria develop an effective mechanism for degradation of microplastics and beads that enables them to utilize plastics as a source of energy without the need for pre-treatments, which highlights the importance of these soil bacteria for the future of effective plastic waste management in a soil environment.

COVID-19 severity: Studying the clinical and demographic risk factors for adverse outcomes.

BACKGROUND: The primary goal of the presented cross-sectional observational study was to determine the clinical and demographic risk factors for adverse coronavirus disease 2019 (COVID-19) outcomes in the Pakistani population. METHODS: We examined the individuals (n = 6331) that consulted two private diagnostic centers in Lahore, Pakistan, for COVID-19 testing between May 1, 2020, and November 30, 2020. The attending nurse collected clinical and demographic information. A confirmed case of COVID-19 was defined as having a positive result through real-time reverse transcriptase polymerase chain reaction (RT-PCR) assay of nasopharyngeal swab specimens. RESULTS: RT-PCR testing was positive in 1094 cases. Out of which, 5.2% had severe, and 20.8% had mild symptoms. We observed a strong association of COVID-19 severity with the number and type of comorbidities. The severity of the disease intensified as the number of comorbidities increased. The most vulnerable groups for the poor outcome are patients with diabetes and hypertension. Increasing age was also associated with PCR positivity and the severity of the disease. CONCLUSIONS: Most cases of COVID-19 included in this study developed mild symptoms or were asymptomatic. Risk factors for adverse outcomes included older age and the simultaneous presence of comorbidities.

Bursting the Virulence Traits of MDR Strain of Candida albicans Using Sodium Alginate-based Microspheres Containing Nystatin-loaded MgO/CuO Nanocomposites.

INTRODUCTION: Candida albicans is a major opportunistic pathogen that causes a wide range of human infections. Currently available therapeutic agents are limited for treating these fungal infections due to multidrug resistance as well as their nonbiodegradability, poor biocompatibility and toxicity. In order to battle these limitations, we have synthesized a polymeric system as microcarriers to deliver the antifungal drug. The objective of the present study was to immobilize MgO/CuO nanocomposite and nystatin-loaded MgO/CuO nanocomposites in nontoxic, nonimmunogenic, biodegradable and biocompatible sodium alginate microspheres for the first time. MATERIALS AND METHODS: Nanoparticle-loaded sodium alginate microspheres were prepared by ionotropic gelation technique using calcium chloride as a cross-linker. Synthesized microspheres were characterized using standard characterization techniques and were evaluated for biological activity against MDR strain of C. albicans. RESULTS: Characterization of microspheres by Fourier-transform infrared spectroscopy confirmed loading of Nys-MgO/CuO NPs, scanning electron microscopy (SEM) revealed rough spherical beads with a highly porous surface having an average size in the range of 8-10 µm. X-ray diffraction (XRD) analyzed its semicrystalline structure. Entrapment efficiency of Nys-MgO/CuO NPs was 80% and release kinetic study revealed sustained and prolonged release of drug in pH 5.5. Flow cytometry analysis showed yeast cell death caused by Nys-MgO/CuO MS exhibits late apoptotic features. In cytotoxicity assay 5-14 mg of microspheres did not cause hemolysis. Microspheres reduced virulence traits of C. albicans such as germ tube and biofilm formation were compromised at concentration of 5 mg/mL. Antimicrobial assessment results revealed a pronounced inhibitory effect against C. albicans. CONCLUSION: The in vitro experiments have shown promising results based on good stability, Nys-MgO/CuO NP-encapsulated microspheres can be used as a prolonged controlled release system against MDR pathogenic C. albicans.

Novel N-p-carboxy benzyl chitosan/poly (vinyl alcohol/functionalized zeolite mixed matrix membranes for DMFC applications.

The novel N-p-carboxy benzyl chitosan (CBC)/ poly (vinyl alcohol) (PVA) based mixed matrix membranes (MMMs) filled with surface-modified zeolite have been prepared using the dissolution casting technique. The applicability of prepared MMMs for direct methanol fuel cell (DMFC) was investigated in terms of water uptake, methanol permeation, and proton conductivity by changing filler content (10-50 wt. %). The zeolite was modified by silane coupling agent, 3-mercaptopropyltrimethoxysilane (MPTMS). The resultant modified zeolite (MZ) was incorporated into CBC/PVA blend to obtain mixed matrix PEMs. The functional group, structural properties, morphological and topographical investigation of MMMs were examined using Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), and Scanning electron microscopy (SEM) respectively. The prepared MMMs exhibited a remarkable decrease in methanol permeability of 2.3 × 10-7  cm2/s with C-CPMZ50. The maximum value of proton conductivity of 0.0527 Scm-1, was shown by C-CMPZ10. The prepared PEMs also displayed good stability during long term operating time.

Antilisterial Effect of Rosa damascena and Nymphaea alba in Mus musculus.

The present study was proposed to investigate the toxicological and prophylactic potential of ethanolic extracts of Rosa damascena and Nymphaea alba and their mixture in albino mice. For toxicity study, three different doses of plant extracts were orally administrated to three groups of mice for 14 successive days. Blood biochemistry and histological examinations of liver and kidney revealed that these extracts had no harmful effects up to 1000 mg/kg. To determine the prophylactic effects of Rosa damascena, Nymphaea alba, and their mixture, an infection model of Listeria monocytogenes was established in a pilot study. Establishment of infection was confirmed by changes in haematological parameters and reisolation of Listeria monocytogenes from different tissues. Results showed that these extracts alone or in combination could restrict the growth of Listeria monocytogenes in different organs. Neutrophils were high in positive control group but remained in normal range in all treated groups. Listeria monocytogenes was recovered in low numbers from animals treated with extract of single plant but was negligible in group treated with mixture of extract of plants. Platelets count was increased in treated groups as compared to control. Results confirmed that these extracts are potent source of antimicrobial compounds and that they have synergistic effect in combined form.

Phytochemical screening and antibacterial potential of Artemisia absinthium L., Swertia chirayita and Sphaeranthus indicus.

Utilization of herbs for medicinal purpose started in the early history of mankind several thousand years ago. In this study, some plants that are used for lowering cholesterol level in local areas of Pakistan, such as Artemisia absinthium L., Swertia chirayita and Sphaeranthus indicus were screened for their phytochemical and antibacterial properties. For this purpose, these plants were extracted in different solvents i.e. ethanol, hexane and ethyl acetate. Phytochemcial analysis unveiled the existence of different bioactive compounds in these extracts. Presence of sugars was further confirmed by performing TLC. Antibacterial activity was determined against indicated bacterial strains, among all extracts Gul-e-mundi had maximum inhibition zone (23mm). DPPH free radical assay revealed the significant antioxidative potential of all the extracts where Gul-e-mundi showed maximum potential i.e., 83%. Plant extracts were also showing anti-proliferative activity on root tips of Allium cepa and Gul-e-mundi was observed to have maximum antimitotic activity i.e. 5%. GC-MS analysis revealed that oleic acid and linoleic acid were the compounds responsible for imparting antibacterial potential to Gul-e-mundi. In conclusion, among all the tested extracts Gul-e-mundi had maximum antibacterial, antioxidative and antimitotic potential. For future studies, phytochemcials responsible for these activities can be isolated and modified for pharmacological purpose.

Production of EPS under Cr(VI) challenge in two indigenous bacteria isolated from a tannery effluent.

Indigenous Cr(VI) reducing bacterial strains Pseudomonas aeruginosa Rb-1 and Ochrobactrum intermedium Rb-2 were evaluated for EPS production under Cr(VI) challenged and free conditions. Strain Rb-2 was more efficient in total EPS production (13.63 mg g(-1)) than Rb-1 (4.15 mg g(-1)) under Cr(VI) stress. Thick covering of capsular material around the cells of both bacterial strains was detected by electron microscopy. Transmission electron micrographs showed the appearance of pilli like structures under chromium stress by two bacteria suggested the possible involvement of this in exchange of hereditary material to increase their chances of survival under stress conditions. FTIR study showed involvement of sulphonate and hydroxyl groups in the binding with Cr(VI) ions. Solid-state (13) C NMR spectra revealed that EPS produced by both strains exhibited structural similarity with the glucan. The partial psl gene sequences of Rb-1 and Rb-2 showed homology with psl gene of Pseudomonas aeruginosa PAO1 and capsular polysaccharide biosynthesis protein of various strains of Pseudomonas. This is the first report on the identification of psl gene from Ochrobacterum in NCBI GenBank database up to our knowledge.

Impact of environmental stress on biochemical parameters of bacteria reducing chromium.

Chromium pollution is produced in connection with industrial processes like in tanneries. It has been suggested that bioremediation could be a good option for clean up. The stress effect of variable chromate levels, pHs and growth temperatures on biochemical parameters of two Cr(VI) reducing bacterial strains Pseudomonas aeruginosa Rb-1 and Ochrobactrum intermedium Rb-2 was investigated. Transmission electrone microscopy (TEM) was performed to study the intracellular distribution of Cr(VI). It was observed that initial stress of 1000 µgmL(-1) caused significant enhancement of all studied biochemical parameters at pH 7.0 and growth temperature of 37 °C showing great bioremediation potential of the strains. Transmission electron microscopy revealed that the distribution of chromium precipitates was not uniform as they were distributed in the cytoplasm as well as found associated with the periplasm and outer membrane. Fourier transform infrared spectroscopy showed the possible involvement of carboxyl, amino, sulpohonate and hydroxyl groups present on the bacterial cell surface for the binding of Cr(VI) ions. Cr(VI) stress brought about changes in the distridution of these functional groups. It can be concluded that the investigated bacterial strains adjust well to Cr(VI) stress in terms of biochemical parameters and along that exhibited alteration in morphology.

Impact of environmental stress on biochemical parameters of bacteria reducing chromium

Chromium pollution is produced in connection with industrial processes like in tanneries. It has been suggested that bioremediation could be a good option for clean up. The stress effect of variable chromate levels, pHs and growth temperatures on biochemical parameters of two Cr(VI) reducing bacterial strains Pseudomonas aeruginosa Rb-1 and Ochrobactrum intermedium Rb-2 was investigated. Transmission electrone microscopy (TEM) was performed to study the intracellular distribution of Cr(VI). It was observed that initial stress of 1000 µgmL-1 caused significant enhancement of all studied biochemical parameters at pH 7.0 and growth temperature of 37 °C showing great bioremediation potential of the strains. Transmission electron microscopy revealed that the distribution of chromium precipitates was not uniform as they were distributed in the cytoplasm as well as found associated with the periplasm and outer membrane. Fourier transform infrared spectroscopy showed the possible involvement of carboxyl, amino, sulpohonate and hydroxyl groups present on the bacterial cell surface for the binding of Cr(VI) ions. Cr(VI) stress brought about changes in the distridution of these functional groups. It can be concluded that the investigated bacterial strains adjust well to Cr(VI) stress in terms of biochemical parameters and along that exhibited alteration in morphology.

Hexavalent chromium reduction by bacteria from tannery effluent.

Chromium is generated from several industrial processes. It occurs in different oxidation states, but Cr(III) and Cr(VI) are the most common ones. Cr(VI) is a toxic, soluble environmental contaminant. Some bacteria are able to reduce hexavalent chromium to the insoluble and less toxic Cr(III), and thus chromate bioremediation is of considerable interest. An indigenous chromium-reducing bacterial strain, Rb-2, isolated from a tannery water sample, was identified as Ochrobactrum intermedium, on the basis of 16S rRNA gene sequencing. The influence of factors like temperature of incubation, initial concentration of Cr, mobility of bacteria, and different carbon sources were studied to test the ability of the bacterium to reduce Cr(VI) under variable environmental conditions. The ability of the bacterial strain to reduce hexavalent chromium in artificial and industrial sewage water was evaluated. It was observed that the mechanism of resistance to metal was not due to the change in the permeability barrier of the cell membrane, and the enzyme activity was found to be inductive. Intracellular reduction of Cr(VI) was proven by reductase assay using cell-free extract. Scanning electron microscopy revealed chromium precipitates on bacterial cell surfaces, and transmission electron microscopy showed the outer as well as inner distribution of Cr(VI). This bacterial strain can be useful for Cr(VI) detoxification under a wide range of environmental conditions.