Harnessing the power of bacterial laccases for xenobiotic degradation in water: A 10-year overview.

Industrialization and population growth are leading to the production of significant amounts of sewage containing hazardous xenobiotic compounds. These compounds pose a threat to human and animal health, as well as the overall ecosystem. To combat this issue, chemical, physical, and biological techniques have been used to remove these contaminants from water bodies affected by human activity. Biotechnological methods have proven effective in utilizing microorganisms and enzymes, particularly laccases, to address this problem. Laccases possess versatile enzymatic characteristics and have shown promise in degrading different xenobiotic compounds found in municipal, industrial, and medical wastewater. Both free enzymes and crude enzyme extracts have demonstrated success in the biotransformation of these compounds. Despite these advancements, the widespread use of laccases for bioremediation and wastewater treatment faces challenges due to the complex composition, high salt concentration, and extreme pH often present in contaminated media. These factors negatively impact protein stability, recovery, and recycling processes, hindering their large-scale application. These issues can be addressed by focusing on large-scale production, resolving operation problems, and utilizing cutting-edge genetic and protein engineering techniques. Additionally, finding novel sources of laccases, understanding their biochemical properties, enhancing their catalytic activity and thermostability, and improving their production processes are crucial steps towards overcoming these limitations. By doing so, enzyme-based biological degradation processes can be improved, resulting in more efficient removal of xenobiotics from water systems. This review summarizes the latest research on bacterial laccases over the past decade. It covers the advancements in identifying their structures, characterizing their biochemical properties, exploring their modes of action, and discovering their potential applications in the biotransformation and bioremediation of xenobiotic pollutants commonly present in water sources.

Microbial host engineering for sustainable isobutanol production from renewable resources.

Due to the limited resources and environmental problems associated with fossil fuels, there is a growing interest in utilizing renewable resources for the production of biofuels through microbial fermentation. Isobutanol is a promising biofuel that could potentially replace gasoline. However, its production efficiency is currently limited by the use of naturally isolated microorganisms. These naturally isolated microorganisms often encounter problems such as a limited range of substrates, low tolerance to solvents or inhibitors, feedback inhibition, and an imbalanced redox state. This makes it difficult to improve their production efficiency through traditional process optimization methods. Fortunately, recent advancements in genetic engineering technologies have made it possible to enhance microbial hosts for the increased production of isobutanol from renewable resources. This review provides a summary of the strategies and synthetic biology approaches that have been employed in the past few years to improve naturally isolated or non-natural microbial hosts for the enhanced production of isobutanol by utilizing different renewable resources. Furthermore, it also discusses the challenges that are faced by engineered microbial hosts and presents future perspectives to enhancing isobutanol production. KEY POINTS: • Promising potential of isobutanol to replace gasoline • Engineering of native and non-native microbial host for isobutanol production • Challenges and opportunities for enhanced isobutanol production.

Characteristics and potential clinical applications of the extracellular vesicles of human pathogenic Fungi.

Extracellular vesicles (EVs) are a heterogeneous group of lipid membrane-enclosed compartments that contain different biomolecules and are released by almost all living cells, including fungal genera. Fungal EVs contain multiple bioactive components that perform various biological functions, such as stimulation of the host immune system, transport of virulence factors, induction of biofilm formation, and mediation of host-pathogen interactions. In this review, we summarize the current knowledge on EVs of human pathogenic fungi, mainly focusing on their biogenesis, composition, and biological effects. We also discuss the potential markers and therapeutic applications of fungal EVs.

Mixed convection and activation energy impacts on MHD bioconvective flow of nanofluid with irreversibility assessment.

In this communication irreversibility minimization in bio convective Walter's-B nanofluid flow by stretching sheet is studied. Suspended nanoparticles in Walter's-B fluid are stabilized by utilizing microorganisms. Total irreversibility is obtained via thermodynamics second law. The influences of applied magnetic field, radiation, Joule heating and activation energy are accounted in momentum, temperature and concentration equations. Furthermore thermophoresis and Brownian movement impacts are also accounted in concentration and temperature expressions. The flow governing dimensional equations are altered into dimensionless ones adopting transformation procedure. Homotopy Analysis Method (HAM) code in Mathematica is implemented to get the convergent series solution. The influences of important flow variables on temperature, velocity, motile density, irreversibility, mass concentration, Bejan number and physical quantities are analyzed graphically. The obtained results revel that the velocity profile decreases for escalating magnetic parameter and Forchheimer number. Entropy generation is increased for higher Brinkman variable while Bejan number declines versus Brinkman variable. The important observations are given at the end.

An Insight into the lynch Syndrome: Retrospective Study of the Pattern of Presentation and Management of Lynch Syndrome in Pakistan.

Introduction: The primary objective of this study was to evaluate the baseline characteristics of Lynch syndrome (LS). Furthermore, the study aimed to evaluate overall survival (OS) among patients with LS. Materials and Methods: This was a retrospective study of colorectal cancer patients registered from January 2010 to August 2020 with an immunohistochemical diagnosis of LS. Results: A total of 42 patients were assessed. The mean age at presentation was 44 years, with male predominance (78%). Demographic preponderance was from the North of Pakistan (52.4%). The family history was positive in 32 (76.2%) patients. The colonic cancer distribution was 32 (76.2%) on the right side. Most of the patients presented with Stage II disease (52.4%), and the common mutations were MLH1 + PMS2 16 (38.1%) followed by MSH2 + MSH6 9 (21.4%). The 10-year OS was found to be 88.1%. However, the OS was 100% post pancolectomy. Conclusion: LS is prevalent in the Pakistan population, especially in the North of Pakistan. Clinical presentation and survivals are similar to the Western population.

Polyarteritis nodosa: a case report of isolated large bowel involvement and surgical intervention.

Polyarteritis nodosa (PAN) is a necrotizing vasculitis commonly affecting small-sized vessels and medium-sized vessels. We present the case of surgical resection for ischemic colitis secondary to PAN. A 26-year-old woman presented with left lower quadrant pain, diarrhea and bloody stools. Colonoscopy showed diffuse circumferential ulceration and necrotic lining from the sigmoid colon to the splenic flexure. A subsequent computed tomography angiography (CTA) revealed left colonic wall thickening, hypoenhancement and multiple inferior mesenteric artery aneurysms, concerning for vasculitis. Ensuing laboratory workup all returned negative. Worsening pain, left-sided peritonitis, tachycardia and rising white blood cell necessitated a colectomy from the proximal descending to upper rectum with an end colostomy. Surgical pathology showed features consistent with PAN. The patient improved significantly after surgery and was discharged on a steroid taper and later started on mycophenolate mofetil. This case is a unique presentation of isolated large intestinal involvement and was treated with both medical and surgical interventions.

CRISPR-Cas9-Based Technology and Its Relevance to Gene Editing in Parkinson's Disease.

Parkinson's disease (PD) and other chronic and debilitating neurodegenerative diseases (NDs) impose a substantial medical, emotional, and financial burden on individuals and society. The origin of PD is unknown due to a complex combination of hereditary and environmental risk factors. However, over the last several decades, a significant amount of available data from clinical and experimental studies has implicated neuroinflammation, oxidative stress, dysregulated protein degradation, and mitochondrial dysfunction as the primary causes of PD neurodegeneration. The new gene-editing techniques hold great promise for research and therapy of NDs, such as PD, for which there are currently no effective disease-modifying treatments. As a result, gene therapy may offer new treatment options, transforming our ability to treat this disease. We present a detailed overview of novel gene-editing delivery vehicles, which is essential for their successful implementation in both cutting-edge research and prospective therapeutics. Moreover, we review the most recent advancements in CRISPR-based applications and gene therapies for a better understanding of treating PD. We explore the benefits and drawbacks of using them for a range of gene-editing applications in the brain, emphasizing some fascinating possibilities.

Endolysin, a Promising Solution against Antimicrobial Resistance.

Antimicrobial resistance (AMR) is a global crisis for human public health which threatens the effective prevention and control of ever-increasing infectious diseases. The advent of pandrug-resistant bacteria makes most, if not all, available antibiotics invalid. Meanwhile, the pipeline of novel antibiotics development stagnates, which prompts scientists and pharmacists to develop unconventional antimicrobials. Bacteriophage-derived endolysins are cell wall hydrolases which could hydrolyze the peptidoglycan layer from within and outside of bacterial pathogens. With high specificity, rapid action, high efficiency, and low risk of resistance development, endolysins are believed to be among the best alternative therapeutic agents to treat multidrug resistant (MDR) bacteria. As of now, endolysins have been applied to diverse aspects. In this review, we comprehensively introduce the structures and activities of endolysins and summarize the latest application progress of recombinant endolysins in the fields of medical treatment, pathogen diagnosis, food safety, and agriculture.

Reliability of modified functional reach test in the assessment of balance function in people with spinal cord injury: A systematic review.

OBJECTIVE: To systematically review studies reporting reliability of modified functional reach test for the assessment of sitting balance function in people with spinal cord injury. METHODS: The systematic review was conducted in line with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines, and comprised research studies published in English language from the earliest record till October 2019 on the subject of reliability of modified functional reach test in assessing balance function in adult spinal cord injury patients.. Prominent databases were searched with relevant key words to shortlist the targeted studies. RESULTS: Of the 108 studies retrieved initially, 6(5.55%) were included in the current systematic review. All the 6(100%) studies reported only test-retest reliability of modified functional reach test. Of them, 4(66.6%) studies measured only forward reach, while 2(33.3%) measured reach in different directions. All 6(100%) studies reported good to excellent reliability of modified functional reach test with interclass coefficient values ranging from 0.78 to 0.99. CONCLUSIONS: Modified functional reach test was found to be a reliable tool for assessing sitting balance function in individuals with spinal cord injury.

Effect of balance training with Biodex Balance System to improve balance in patients with diabetic neuropathy: A quasi experimental study.

OBJECTIVE: To determine the effect of balance training with Biodex Balance System in improving balance function in patients with diabetic neuropathy. METHODS: A quasi experimental study was conducted at physiotherapy department of Rehman Medical Institute Peshawar from January to June 2018. Thirty-eight Diabetes Mellitus Type-II patients with diagnosis of diabetic neuropathy participated in the study. Those patients were included who were able to stand independently and who scored moderate or severe neuropathy on Toronto Scale. Participants received 12 sessions (two sessions per week) of balance training on Biodex stability system in six weeks. Berg Balance Scale and Time Up and Go test were used to collect pre and post treatment data. Paired sample T-test was used to compare pre and post treatment data because data was normally distributed. RESULTS: The mean age of the participants was 63.08 ± 3.3 years. Pretreatment Berg Balance Scale score was 42.68 ± 3.7 which improved to 48.89 ±3.7 after the treatment (P<0.001). Similarly, pretreatment Time Up and Go test score was 17.47 ± 2.0 while post treatment score was 13.57 ±1.5 (P<0.001). CONCLUSION: Balance training with Biodex Balance System can significantly improve balance function in Diabetes Mellitus Type-II patients with diabetic neuropathy.

Assembly properties of bacterial tubulin homolog FtsZ regulated by the positive regulator protein ZipA and ZapA from Pseudomonas aeruginosa.

Bacterial tubulin homolog FtsZ self-assembles into dynamic protofilaments, which forms the scaffold for the contractile ring (Z-ring) to achieve bacterial cell division. Here, we study the biochemical properties of FtsZ from Pseudomonas aeruginosa (PaFtsZ) and the effects of its two positive regulator proteins, ZipA and ZapA. Similar to Escherichia coli FtsZ, PaFtsZ had a strong GTPase activity, ~ 7.8 GTP min-1 FtsZ-1 at pH 7.5, and assembled into mainly short single filaments in vitro. However, PaFtsZ protofilaments were mixtures of straight and "intermediate-curved" (100-300 nm diameter) in pH 7.5 solution and formed some bundles in pH 6.5 solution. The effects of ZipA on PaFtsZ assembly varied with pH. In pH 6.5 buffer ZipA induced PaFtsZ to form large bundles. In pH 7.5 buffer PaFtsZ-ZipA protofilaments were not bundled, but ZipA enhanced PaFtsZ assembly and promoted more curved filaments. Comparable to ZapA from other bacterial species, ZapA from P. aeruginosa induced PaFtsZ protofilaments to associate into long straight loose bundles and/or sheets at both pH 6.5 and pH 7.5, which had little effect on the GTPase activity of PaFtsZ. These results provide us further information that ZipA functions as an enhancer of FtsZ curved filaments, while ZapA works as a stabilizer of FtsZ straight filaments.

Effectiveness of manual therapy to the cervical spine with and without manual therapy to the upper thoracic spine in the management of non-specific neck pain; a randomized controlled trial.

OBJECTIVES: To compare the effectiveness of manual therapy to the cervical spine with and without manual therapy to the upper thoracic spine in the management of non-specific neck pain. METHODS: The randomized controlled trial was conducted at 3 different hospitals in Peshawar, Pakistan, from October 2016 to January 2017, and comprised patients suffering from non-specific neck pain aged 25-60 years. The control group received cervical manual therapy alone while the experimental group received cervical along with thoracic manual therapy for 2 weeks. Data was analysed using SPSS 20. RESULTS: Of the 37 subjects, 20(54%) were cases and 17(46%) were controls.The overall mean age was 35.9±9.6 years. There was no significant difference between the groups at baseline in terms of the levels of pain (p=0.125) and disability (p=0.392). The experimental group showed greater reduction in pain (p=0.02) and disability (p=0.03) compared to the control group. CONCLUSIONS: Cervical along with thoracic manual therapy reduced neck pain and associated neck disability more effectively than cervical manual therapy alone.

Evaluation of entropy generation in cubic autocatalytic unsteady squeezing flow of nanofluid between two parallel plates.

BACKGROUND: Nanomaterials have advanced behaviors that make them possibly beneficial in various applications in mass and heat transports such as engine cooling, pharmaceutical processes, fuel cells, engine cooling and domestic refrigerator etc. Therefore here we deliberated the entropy generation in unsteady magnetohydrodynamic squeezing flow of viscous nanomaterials between two parallel plates. The upper plate is squeezing towards lower plate. The lower plate exhibits porous character. Energy attributes are discussed through heat flux, dissipation and Joule heating. Furthermore the irreversibility analysis with cubic autocatalysis chemical reaction is also accounted. METHODS: Nonlinear differential systems are converted to ordinary differential system by transformations. For convergent series solution the given system are solved by homotopy analysis method (HAM). RESULTS: Characteristics of various interesting variables on velocity, Bejan number, concentration, entropy optimization and temperature are deliberated through graphs. Gradient of velocity (Cfx) and Nusselt number (Nux) are numerically computed against various physical variables. Entropy generation and Bejan number both quantitatively enhance versus radiation parameter. For larger squeezing parameter the velocity and temperature field are increased. CONCLUSIONS: The obtained results show that for larger squeezing parameter the velocity field boosts up. Velocity have opposite impact For larger magnetic and porosity parameters. Temperature is decreased for higher values of radiation parameter and Prandtl number. Temperature and concentration have same outcome for thermophoresis parameter. Entropy generation and Bejan number both quantitatively enhance versus radiation parameter, while reverse is hold for Brinkman number.

Assembly properties of the bacterial tubulin homolog FtsZ from the cyanobacterium Synechocystis sp. PCC 6803.

The tubulin homolog FtsZ is the major cytoskeletal protein in the bacterial cell division machinery, conserved in almost all bacteria, archaea, and chloroplasts. Bacterial FtsZ assembles spontaneously into single protofilaments, sheets, and bundles in vitro, and it also accumulates at the site of division early during cell division, where it forms a dynamic protein complex, the contractile ring or Z-ring. The biochemical properties of FtsZ proteins from many bacteria have been studied, but comparable insights into FtsZs from cyanobacteria are limited. Here, using EM and light-scattering assays, we studied the biochemical and assembly properties of SyFtsZ, the FtsZ protein from the cyanobacterial strain Synechocystis sp. PCC 6803. SyFtsZ had a slow GTPase activity of ∼0.4 GTP/FtsZ molecule/min and assembled into thick, straight protofilament bundles and curved bundles, designated toroids. The assembly of SyFtsZ in the presence of GTP occurred in two stages. The first stage consisted of the assembly of single-stranded straight protofilaments and opened circles; in the second stage, the protofilaments associated into straight protofilament bundles and toroids. In addition to these assemblies, we also observed highly curved oligomers and minirings after GTP hydrolysis or in the presence of excess GDP. The three types of protofilaments of SyFtsZ observed here provide support for the hypothesis that a constriction force due to curved protofilaments bends the membrane. In summary, our findings indicate that, unlike other bacterial FtsZ, SyFtsZ assembles into thick protofilament bundles. This bundling is similar to that of chloroplast FtsZ, consistent with its origin in cyanobacteria.

Similar virulence properties of infection and colonization associated Pseudomonas aeruginosa.

PURPOSE: Pseudomonas aeruginosa is one of the agents that are commonly implicated in nosocomial infections. However, it is also present as a commensal in various body sites of healthy persons, making the diagnosis of infection by culture difficult. A number of virulence factors expressed by the organism have been implicated in its pathogenicity. We undertook this study to identify the host and organism factors associated with infection. METHODOLOGY: Pathogenic, colonizing and environmental isolates were tested for apr, lasB, the T3SS effector exoenzymes (exoS, exoT, exoU and exoY) and toxA genes, biofilm production and antimicrobial susceptibility. The isolates were further typed by RAPD. RESULTS: Eighty-seven isolates from 61 patients, including 11 environmental isolates, were obtained. None of the virulence factors were found to be significantly associated with infection, and nor was the antimicrobial susceptibility. The presence of the exoU gene and infection by MDR strains correlated significantly with the duration of hospital stay. Positivity for exoS and exoU genes was found to be strongly correlated with multi-drug resistance. exoU positivity correlated strongly with fluoroquinolone resistance. Sinks in the ward and intensive care unit were found to be a niche for XDR P. aeruginosa. Eighty-five isolates were typeable using the ERIC2 primer, showing 71 distinct RAPD patterns with >15 % difference in UPGMA-generated dice coefficients. CONCLUSIONS: exoU positivity is associated with severe disease, as evidenced by the longer duration of hospital stay of these patients. However, the presence of virulence factors or multi-drug resistance in the cultured strain should not prompt the administration of anti-pseudomonal chemotherapy.

Prediction equations for spirometry in adults in western India.

BACKGROUND: Spirometry is an essential investigation in pulmonology. The predicted normal spirometry values depend on various physiological parameters. This study was conducted to collect updated information on pulmonary functions in normal adults from western India. MATERIAL AND METHODS: A prospective observational study was undertaken at a tertiary hospital in Mumbai enrolling healthy subjects, 18-75 years, with ethnic origin from western India. Spirometry measurements were carried out as per ATS/ERS-2005 guidelines using a non-heated Fleish Pneumotachograph spirometer. Data was analyzed using SPSS for Pearson's correlation analysis, multiple linear regressions and log transformations of variables to get the best prediction equations. RESULTS: 310 subjects (185 males, 125 females) were included. Lung function values were higher in men as compared to women. In multivariate linear regression models, age and height were major predictor variables for all spirometry parameters. Addition of weight as a determinant variable did not make significant contribution to the models except for PEFR in males and F75 in females. Regression equations were established for FVC, FEV1, FEV1/FVC ratio, PEFR, F25-75, F50, and F75. The standard-error-of-estimate was provided to enable computation of lower limits of normal for these parameters. CONCLUSION: We propose regression equations for spirometry variables developed using the current standards for adult West Indian population fulfilling the long-felt need for updated equations.

Design and synthesis of benzimidazole analogs endowed with oxadiazole as selective COX-2 inhibitor.

New molecules of benzimidazole endowed with oxadiazole were designed and synthesized from 2-(2-((pyrimidin-2-ylthio)methyl)-1H-benzo[d]imidazol-1-yl)acetohydrazide as 1-((5-substituted alkyl/aryl-1,3,4-oxadiazol-2-yl)methyl)-2-((pyrimidin-2-ylthio)methyl)-1H-benzimidazoles (5a-r) with the aim to acquire selective cyclooxygenase (COX-2) inhibitor activity. The synthesized compounds were screened by in vitro cyclooxygenase assays to determine COX-1 and COX-2 inhibitory potency and the results showed that they had good-to-remarkable activity with an IC50 range of 11.6-56.1 µM. The most active compounds were further screened for their in vivo anti-inflammatory activity by using the carrageenan-induced rat paw edema model. In vitro anticancer activities of the hybrid compounds were assessed by the National Cancer Institute (NCI), USA, against 60 human cell lines, and the results showed a good spectrum. Compound 5l exhibited significant COX-2 inhibition with an IC50 value of 8.2 µM and a percent protection of 68.4%. Compound 5b evinced moderate cytotoxicity toward the UO-31 cell line of renal cancer. A docking study was performed using Maestro 9.0, to provide the binding mode into the binding sites of the cyclooxygenase enzyme. Hopefully, in the future, compound 5l could serve as a lead compound for developing new COX-2 inhibitors.

Synthesis and antihypertensive screening of new derivatives of quinazolines linked with isoxazole.

A series of 7-substituted-3-(4-(3-(4-substitutedphenyl)-4,5-dihydroisoxazol-5-yl)phenyl)-2-substituted quinazolin-4(3H)-one (1-30) have been synthesized by the cyclization of (E)-3-(4-(3-substitutedphenyl)acrylolyl)phenyl)-2-(substitutedphenyl)-7-substituted quinazolin-4-(3H)-one with hydroxylamine hydrochloride. The synthesized compounds were examined for their in vivo antihypertensive activity using albino rats. All the titled compounds exhibited good to moderate antihypertensive activity. Compounds 7-Chloro-3-(4-(3-(4-chlorophenyl)-4,5- dihydroisoxazol-5-yl)phenyl)-2-p-tolylquinazolin-4(3H)-one (23) and 7-Chloro-3-(4-(3-(4-chlorophenyl)-4,5-dihydroisoxazol-5-yl)phenyl)-2-(4-methoxyphenyl)quinazolin-4(3H)-one (24) exhibited potent antihypertensive activity through their anticipated α 1-adrenergic receptor blocking property similar to its clinically used analogue, prazosin, without affecting heart rate with prolonged duration of action when tested in adrenaline induced hypertension in anaesthetized rats.

Estimation of absorbed dose to thyroid in patients treated with radiotherapy for various cancers.

This study is aimed at the evaluation of the absorbed dose to the thyroid gland from scattered/primary radiations in patients who have undergone radiotherapy. In this study, the skin entrance dose (SED) on the anterior neck at the level of the thyroid gland was measured using thermoluminescent dosimeters for 57 patients during radiotherapy with (60)Co teletherapy unit. Phantom dosimetry was also performed to find the correlation between the SED and the dose at the depth of the thyroid. SED of patients treated for breast cancer was 6.77±3.49 Gy and that of those with Hodgkin's disease and Ca. lung irradiation were 21.29±13.52 Gy and 28.80±14.94 Gy, respectively. SED at the level of the thyroid gland was found to be highest for patients whose thyroid gland was within the radiation field, while lowest for patients with skull irradiation. Thyroid gland is recommended to be shielded or excluded from the field whenever possible.