Esterases as emerging biocatalysts: Mechanistic insights, genomic and metagenomic, immobilization, and biotechnological applications.

Esterase enzymes are a family of hydrolases that catalyze the breakdown and formation of ester bonds. Esterases have gained a prominent position in today's world's industrial enzymes market. Due to their unique biocatalytic attributes, esterases contribute to environmentally sustainable design approaches, including biomass degradation, food and feed industry, dairy, clothing, agrochemical (herbicides, insecticides), bioremediation, biosensor development, anticancer, antitumor, gene therapy, and diagnostic purposes. Esterases can be isolated by a diverse range of mammalian tissues, animals, and microorganisms. The isolation of extremophilic esterases increases the interest of researchers in the extraction and utilization of these enzymes at the industrial level. Genomic, metagenomic, and immobilization techniques have opened innovative ways to extract esterases and utilize them for a longer time to take advantage of their beneficial activities. The current study discusses the types of esterases, metagenomic studies for exploring new esterases, and their biomedical applications in different industrial sectors.

Ethyl-acetate extract of tara mira (Eruca sativa) alleviates the inflammation and rheumatoid arthritis in rats.

Eruca sativa, member of family Brassicaceae, was evaluated for its anti-arthritic potential. Both in vitro and in vivo models were used to bring out a safe, effective and economical remedy. In vitro tests included egg albumin denaturation suppression, bovine serum albumin assay and human red blood cells maintenance assay. While in vivo formaldehyde-induced arthritic model was initiated to check effect on paw volume. Similarly, carrageenan produced inflammation was applied to check anti-inflammatory ability of the plant. Acute toxicity studies showed safety margin at 2000mg/kg. The plant showed concentration dependent denaturation protection and membrane stability in vitro assays. Likewise, the carrageenan and formaldehyde investigations revealed visible paw volume reduction in dose attributed manner, with maximum outcome at dose of 500mg/kg. Hence, it may be established on the ground of presented results that ethyl-acetate extract of Eruca sativa has significant anti-inflammatory and anti-arthritic effects and may be considered for further research to reveal the core mechanism.

Zeta potential changing self-nanoemulsifying drug delivery systems: A newfangled approach for enhancing oral bioavailability of poorly soluble drugs.

The mucus is a defensive barrier for different drug-loaded systems. To overcome this obstacle, the crucial factor is the surface charge. Due to mucus negative charge behavior; it was revealed that negatively charged formulations can move across mucus, whereas positively charged nanoformulations could not diffuse via mucus due to interactions. However, cellular intake of negatively charged nanoformulations to the epithelium by endocytosis is less prominent as compared to positively charged carriers. Self-emulsifying drug delivery systems (SEDDS) improve the drug permeability of drugs, especially which have poor oral drug solubility. Moreover, SEDDS have the ability to reduce the degradation of drugs in the GI tract. Currently, drug carrier systems that can shift zeta potential from negative to positive were developed. The benefits of inducing zeta potential changing approach are that negatively charged nanoformulations permeate quickly across the mucus and surface charges reversed to positive at epithelium surface to increase cellular uptake. Among various systems of drug delivery, zeta potential changing SEDDS seem to signify a promising approach as they can promptly diffuse over mucus due to their smaller size and shape distortion ability. Due to such findings, mucus permeation and drug diffusion may improve by the mixture of the zeta potential changing approach and SEDDS.

Genetically engineered microorganisms for environmental remediation.

In the recent era, the increasing persistence of hazardous contaminants is badly affecting the globe in many ways. Due to high environmental contamination, almost every second species on earth facing the worst issue in their survival. Advances in newer remediation approaches may help enhance bioremediation's quality, while conventional procedures have failed to remove hazardous compounds from the environment. Chemical and physical waste cleanup approaches have been used in current circumstances; however, these methods are costly and harmful to the environment. Thus, there has been a rise in the use of bioremediation due to an increase in environmental contamination, which led to the development of genetically engineered microbes (GEMs). It is safer and more cost-effective to use engineered microorganisms rather than alternative methods. GEMs are created by introducing a stronger protein into bacteria through biotechnology or genetic engineering to enhance the desired trait. Biodegradation of oil spills, halobenzoates naphthalenes, toluenes, trichloroethylene, octanes, xylenes etc. has been accomplished using GEMs such bacteria, fungus, and algae. Biotechnologically induced microorganisms are more powerful than naturally occurring ones and may degrade contaminants faster because they can quickly adapt to new pollutants they encounter or co-metabolize. Genetic engineering is a worthy process that will benefit the environment and ultimately the health of our people.

In-situ, Ex-situ, and nano-remediation strategies to treat polluted soil, water, and air - A review.

Nanotechnology, as an emerging science, has taken over all fields of life including industries, health and medicine, environmental issues, agriculture, biotechnology etc. The use of nanostructure molecules has revolutionized all sectors. Environmental pollution is a great concern now a days, in all industrial and developing as well as some developed countries. A number of remedies are in practice to overcome this problem. The application of nanotechnology in the bioremediation of environmental pollutants is a step towards revolution. The use of various types of nanoparticles (TiO2 based NPs, dendrimers, Fe based NPs, Silica and carbon nanomaterials, Graphene based NPs, nanotubes, polymers, micelles, nanomembranes etc.) is in practice to diminish environmental hazards. For this many In-situ (bioventing, bioslurping, biosparging, phytoremediation, permeable reactive barrier etc.) and Ex-situ (biopile, windrows, bioreactors, land farming etc.) methodologies are employed. Improved properties like nanoscale size, less time utilization, high adaptability for In-situ and Ex-situ use, undeniable degree of surface-region to-volume proportion for possible reactivity, and protection from ecological elements make nanoparticles ideal for natural applications. There are distinctive nanomaterials and nanotools accessible to treat the pollutants. Each of these methods and nanotools depends on the properties of foreign substances and the pollution site. The current designed review highlights the techniques used for bioremediation of environmental pollutants as well as use of various nanoparticles along with proposed In-situ and Ex-situ bioremediation techniques.

Dendritic Cell-Targeted Therapies to Treat Neurological Disorders.

Dendritic cells (DCs) are the immune system's highly specialized antigen-presenting cells. When DCs are sluggish and mature, self-antigen presentation results in tolerance; however, when pathogen-associated molecular patterns stimulate mature DCs, antigen presentation results in the development of antigen-specific immunity. DCs have been identified in various vital organs of mammals (e.g., the skin, heart, lungs, intestines, and spleen), but the brain has long been thought to be devoid of DCs in the absence of neuroinflammation. However, neuroinflammation is becoming more recognized as a factor in a variety of brain illnesses. DCs are present in the brain parenchyma in trace amounts under healthy circumstances, but their numbers rise during neuroinflammation. New therapeutics are being developed that can reduce dendritic cell immunogenicity by inhibiting pro-inflammatory cytokine production and T cell co-stimulatory pathways. Additionally, innovative ways of regulating dendritic cell growth and differentiation and harnessing their tolerogenic capability are being explored. Herein, we described the function of dendritic cells in neurological disorders and discussed the potential for future therapeutic techniques that target dendritic cells and dendritic cell-related targets in the treatment of neurological disorders.

Neurotoxic effects of environmental contaminants-measurements, mechanistic insight, and environmental relevance.

Pollution is a significant and growing concern for any population regardless of age because these environmental contaminants exhibit different neurodegenerative effects on persons of different ages. These environmental contaminants are the products of human welfare projects like industry, automobile exhaust, clinical and research laboratory extrudes, and agricultural chemicals. These contaminants are found in various forms in environmental matrices like nanoparticles, particulate matter, lipophilic vaporized toxicants, and ultrafine particulate matter. Because of their small size, they can easily cross blood-brain barriers or use different cellular mechanisms for assistance. Other than this, these contaminants cause an innate immune response in different cells of the central nervous system and cause neurotoxicity. Considering the above critiques and current needs, this review summarizes different protective strategies based on bioactive compounds present in plants. Various bioactive compounds from medicinal plants with neuroprotective capacities are discussed with relevant examples. Many in vitro studies on clinical trials have shown promising outcomes using plant-based bioactive compounds against neurological disorders.

SARS-COV-2/COVID-19: scenario, epidemiology, adaptive mutations, and environmental factors.

The coronavirus pandemic of 2019 has already exerted an enormous impact. For over a year, the worldwide pandemic has ravaged the whole globe, with approximately 250 million verified human infection cases and a mortality rate surpassing 4 million. While the genetic makeup of the related pathogen (SARS-CoV-2) was identified, many unknown facets remain a mystery, comprising the virus's origin and evolutionary trend. There were many rumors that SARS-CoV-2 was human-borne and its evolution was predicted many years ago, but scientific investigation proved them wrong and concluded that bats might be the origin of SARS-CoV-2 and pangolins act as intermediary species to transmit the virus from bats to humans. Airborne droplets were found to be the leading cause of human-to-human transmission of this virus, but later studies showed that contaminated surfaces and other environmental factors are also involved in its transmission. The evolution of different SARS-CoV-2 variants worsens the condition and has become a challenge to overcome this pandemic. The emergence of COVID-19 is still a mystery, and scientists are unable to explain the exact origin of SARS-CoV-2. This review sheds light on the possible origin of SARS-CoV-2, its transmission, and the key factors that worsen the situation.

Anticancer L-Asparaginase and Phytoactive Compounds From Plant Solanum nigrum Against MDR (Methicillindrug resistant) Staphylococcus aureus and Fungal Isolates.

L-asparaginase is used in chemotherapy for acute lymphoblastic leukemia and other cancers. L-asparaginase derived from bacterial source triggers immune responses. The current study investigates Solanum nigrum as a novel and latent source of L-asparaginase to minimize immunological reactions. The antitumor activity of SN methanol extract was determined using the potato disc assay. InterPro Chimera and InterPro were used to predict the amino acid sequence of L-asparaginase and its anticancer activity. Purification of the enzyme was carried out to homogeneity of 1.51-fold with a recovery of 61.99%. At optimal conditions of 36.5°C, pH 8.6, and 8.5 g/mL substrate, fruit (crude extract) revealed an L-asparaginase titer of 48.23 U/mL. The molecular weight of the enzyme was calculated to be 32 ± 5 kDa using SDS PAGE. The fruit's total flavonoids and phenolic contents are 0.42 ± .030 g/mL and 94 ± 1.9 mg CAE, respectively. Anti-tumorigenic efficacy was determined to be 66% against Agrobacterium tumefaciens. Additionally, the extract possesses potent antifungal and antibacterial properties. Molecular docking provided the structural motifs and underlying interactions between L-asparaginase, N-acetylglucosamine, murine, and chitin. SN contains high levels of the enzyme L-asparaginase and phytochemicals, making it a potential source of anticancer drugs.

Engineered tyrosinases with broadened bio-catalysis scope: immobilization using nanocarriers and applications.

Enzyme immobilization is a widely used technology for creating more stable, active, and reusable biocatalysts. The immobilization process also improves the enzyme's operating efficiency in industrial applications. Various support matrices have been designed and developed to enhance the biocatalytic efficiency of immobilized enzymes. Given their unique physicochemical attributes, including substantial surface area, rigidity, semi-conductivity, high enzyme loading, hyper catalytic activity, and size-assisted optical properties, nanomaterials have emerged as fascinating matrices for enzyme immobilization. Tyrosinase is a copper-containing monooxygenase that catalyzes the o-hydroxylation of monophenols to catechols and o-quinones. This enzyme possesses a wide range of uses in the medical, biotechnological, and food sectors. This article summarizes an array of nanostructured materials as carrier matrices for tyrosinase immobilization. Following a detailed background overview, various nanomaterials, as immobilization support matrices, including carbon nanotubes (CNTs), carbon dots (CDs), carbon black (CB), nanofibers, Graphene nanocomposite, platinum nanoparticles, nano-sized magnetic particles, lignin nanoparticles, layered double hydroxide (LDH) nanomaterials, gold nanoparticles (AuNPs), and zinc oxide nanoparticles have been discussed. Next, applied perspectives have been spotlights with particular reference to environmental pollutant sensing, phenolic compounds detection, pharmaceutical, and food industry (e.g., cereal processing, dairy processing, and meat processing), along with other miscellaneous applications.

Current scenario of COVID-19 vaccinations and immune response along with antibody titer in vaccinated inhabitants of different countries.

The COVID-19 pandemic challenges have been only partially addressed so far. The pathogenicity of SARS-CoV-2 is considered the combination of severe and high infectivity. Herdimmunity is attained when a critical proportion of the population is immune, providing the virus with fewer chances to spread locally. To overcome the rising tide of the COVID-19 pandemic, efficacious and safe vaccines providing defensive and long-lasting immunity responses are urgently needed.Vaccines that induce virus-neutralizing antibodies with great affinity can optimally fight against infection. Worldwide, over 120 novel vaccine candidates, including live-attenuated, inactivated, viral-vectored nonreplicating and replicating, peptide- and protein-based, and nucleic acid-based approaches are in the process of preclinical and clinical trials (phase 1-4). In addition to comprehensive safety assessments and immune responses, precise clinical management is also important for trials of vaccines. The recent emergence of different variants of SARS-CoV-2 is becoming a new threat for the world and a challenge for scientists to introduce the most influential vaccine against COVID-19. The possibility of natural and vaccine-induced immunity in variants finds it necessary to establish next-generation vaccines, which generate general neutralization against existing and future variants. Here, we summarize the cellular and humoral responses of SARS-CoV-2, current progress in vaccination development, the antibody titer response of available phase 4 vaccinations in vaccinated populations of different countries worldwide, and the success and challenges ahead of vaccine development.

Expanding the bio-catalysis scope and applied perspectives of nanocarrier immobilized asparaginases.

l-asparaginase is an essential enzyme in medicine and a well-known chemotherapeutic agent. This enzyme's importance is not limited to its use as an anti-cancer agent; it also has a wide variety of medicinal applications. Antimicrobial properties, prevention of infectious disorders, autoimmune diseases, and canine and feline cancer are among the applications. Apart from the healthcare industry, its importance has been identified in the food industry as a food manufacturing agent to lower acrylamide levels. When isolated from their natural habitats, they are especially susceptible to different denaturing conditions due to their protein composition. The use of an immobilization technique is one of the most common approaches suggested to address these limitations. Immobilization is a technique that involves fixing enzymes to or inside stable supports, resulting in a heterogeneous immobilized enzyme framework. Strong support structures usually stabilize the enzymes' configuration, and their functions are maintained as a result. In recent years, there has been a lot of curiosity and focus on the ability of immobilized enzymes. The nanomaterials with ideal properties can be used to immobilize enzymes to regulate key factors that determine the efficacy of bio-catalysis. With applications in biotechnology, immunosensing, biomedicine, and nanotechnology sectors have opened a realm of opportunities for enzyme immobilization.

S-Protected thiolated nanostructured lipid carriers exhibiting improved mucoadhesive properties.

The purpose of the present study was to design nanostructured lipid carriers (NLCs) exhibiting improved mucoadhesive properties. First, an S-protected thiolated fatty acid conjugate was synthesized by amide bond formation between a primary amino group of l-cystine and palmitic acid N-hydroxysuccinimide. NLCs were prepared by nano-template engineering technique using Span 60, polysorbate 80, sucrose stearate and PEG 400 as surfactant mixture, stearic acid as solid lipid and miglyol as liquid lipid. NLCs were loaded with the model drug bergapten and decorated with the S-protected thiolated fatty acid conjugate. NLCs were characterized regarding particle size, poly-dispersity index (PDI), zeta potential, drug entrapment efficiency (EE), drug loading capacity (LC), drug release and mucoadhesive properties. Furthermore, cytotoxicity studies were performed on MDA-MB-231 cells via resazurin assay. S-Protected thiolated NLCs displayed a mean size of 115 nm, PDI of 0.3, zeta potential of -30 mV, 80% drug EE and 5% drug LC. Drug-loaded S-protected thiolated NLCs exhibited a sustained drug release and strongly enhanced mucoadhesive properties. Surface decoration with cystine substructures raised the cytotoxic potential of NLCs to a minor extent. Due to the immobilization of cystine substructures on the surface of NLCs, their mucoadhesive properties can be strongly improved.

Tetradeca-thiolated cyclodextrins: Highly mucoadhesive and in-situ gelling oligomers with prolonged mucosal adhesion.

The purpose of this study was to synthesize a highly mucoadhesive tetradeca-thiolated ß-cyclodextrin (ß-CD) by replacement of all primary OH groups at C-6 position and all secondary OH groups at C-2 position of ß-CD backbone viaSH groups and to evaluate its rheological and mucoadhesive properties in-vitro. Primary and secondary OH groups of ß-CD were substituted by SH groups using a microwave-assisted method. The structure of tetradeca-thiolated ß-CD was confirmed by FTIR and 1H NMR spectroscopy. The modified ß-CD was evaluated for SH content, thiol stability towards oxidation and cytotoxicity. Moreover, the viscoelastic behavior of the modified oligomer was investigated via rheological studies with porcine intestinal mucus and fibrous structural protein keratin, whereas mucoadhesive properties were evaluated using different porcine mucosae. Tetradeca-thiolated ß-CD oligomer displayed 8144 ± 317 µmol thiol groups per gram. These thiol groups displaying a pKa value of 8.2 were stable at pH 4 but prone to oxidation at higher pH values. The newly synthesized thiolated CD did not show any cytotoxicity to Caco-2 cells at a concentration of 0.5% (m/v) within 24 h. Due to the addition of 0.5 and 2% (m/v) tetradeca-thiolated ß-CD to mucus and keratin, the dynamic viscosity was increased up to 7.6- and 5.9- fold, respectively, within 4 h at 37 °C. Moreover, in-vitro mucoadhesion studies of tetradeca-thiolated CD showed 78.6-, 60.3-, 62.3- and 49.3- fold improved mucoadhesion on intestinal, buccal, bladder and vaginal mucosa as compared to unmodified ß-CD, respectively. According to these results, tetradeca-thiolated ß-CD might be a promising auxiliary agent to provide a prolonged residence time of drug delivery systems on different mucosal surfaces.

Development of pre-activated α-cyclodextrin as a mucoadhesive excipient for intra-vesical drug delivery.

The study was designed to synthesize and characterize pre-activated α-cyclodextrin (α-CD) derivatives as mucus adhering excipients for intra-vesical drug delivery. Sodium periodate (NaIO4) was used to oxidize α-CD and subsequently cysteamine was covalently attached to carbonyl groups of oxidized α-CD via reductive amination to produce thiolated α-CD. l-cysteine-2-mercaptonicotinic acid conjugate (Cys-MNA) was covalently attached to carbonyl groups of oxidized α-CD to produce pre-activated α-CD having enhance stability against oxidation at higher pH. Thiolated and pre-activated α-CD derivatives were quantitatively assayed for the attached thiol groups and MNA groups, respectively. Cell viability and tolerability was evaluated via resazurin assay and via red blood cells (RBC) lysis assay, respectively. Mucoadhesive properties were evaluated on porcine bladder mucosa. Trimethoprim (TMP) was encapsulated into thiolated and pre-activated α-CD derivatives and the dissolution behavior was evaluated in vitro. Thiol groups attached to thiolated α-CD derivatives α-CD-SH780 and α-CD-SH1426 were 780±68µmol/g and 1426±66µmol/g, respectively. For the entirely pre-activated α-CD derivatives, α-CD-MNA3609 and α-CD-MNA4285 number of attached MNA groups were 3609±19µmol/g and 4285±43µmol/g, respectively. Thiolated and pre-activated derivatives of α-CD did not show adverse effects to cells determined via resazurin and RBC lysis assays. Mucoadhesion on porcine bladder mucosa was significantly improved for thiolated and pre-activated α-CD derivatives. Thiolated α-CD-SH1426 showed 15-fold and pre-activated α-CD-MNA4285 showed 25-fold improved mucoadhesion compared to unmodified α-CD. Further, pre-activated α-CD-MNA4285 showed 2-fold enhanced dissolution of encapsulated TMP compared to free TMP over 3 h. The study shows that pre-activated α-CD could be an excipient of the choice for the formulations of mucoadhesive intra-vesical drug delivery systems.

Upper extremity deep vein thrombosis.

A case of deep vein thrombosis of axillary vein (Paget-Schroetter syndrome) is described in an 18 years old male. Patient presented via emergency with a one week history of pain and swelling of arm. Duplex imaging showed deep vein thrombosis of third part of left axillary vein and venae comitantes of brachial artery. Patient's condition resolved on conservative treatment.