A systematic review on enhancement strategies in biochar-based remediation of polycyclic aromatic hydrocarbons.

Polycyclic aromatic hydrocarbons (PAHs) are pervasive ecological pollutants produced essentially during the inadequate burning of organic materials. PAHs are a group of different organic compounds that are made out of various aromatic rings. PAHs pose a serious risk to humans and aquatic ecosystems because of their mutagenic and carcinogenic properties. In this way, there is a critical prerequisite to utilizing successful remediation strategies and methods to limit the dangerous effect of these pollutants on the ecosystem. Biochar has believed of intriguing properties such as simple manufacturing operations and more affordable and more productive materials. Biochar is a sustainable carbonaceous material that has an enormous surface area with bountiful functional groups and pore structure, which has huge potential for the remediation of toxic pollutants. This review emphasizes the occurrence, development, and fate of toxic PAHs in the environment. In the present review, the properties and role of biochar in the removal of PAHs were illustrated, and the influencing factors and an efficient key mechanism of biochar for the remediation of PAHs were discussed in detail. Various surface modification methods can be utilized to improve the biochar properties with the magnetization process; the advancements of modified biochar are pointed out in this review. Finally, the constraints and prospects for the large-scale application of biochar in the remediation of toxic pollutants are highlighted.

Efficiency of various biofilm carriers and microbial interactions with substrate in moving bed-biofilm reactor for environmental wastewater treatment.

In a moving bed-biofilm reactor (MBBR), the fluidization efficiency, immobilization of microbial cells, and treatment efficiency are directly influenced by the shape and pores of biofilm carriers. Moreover, the efficacy of bioremediation mainly depends on their interaction interface with microbes and substrate. This review aims to comprehend the role of different carrier properties such as material shapes, pores, and surface area on bioremediation productivity. A porous biofilm carrier with surface ridges containing spherical pores sizes > 1 mm can be ideal for maximum efficacy. It provides diverse environments for cell cultures, develops uneven biofilms, and retains various cell sizes and biomass. Moreover, the thickness of biofilm and controlled scaling shows a significant impact on MBBR performance. Therefore, the effect of these parameters in MBBR is discussed detailed in this review, through which existing literature and technical strategies that focus on the surface area as the primary factor can be critically assessed.

Strategies for successful designing of immunocontraceptive vaccines and recent updates in vaccine development against sexually transmitted infections - A review.

Background: The world population is continuously growing. It has been estimated that half of the world's population is from the Asian continent, mainly from China and India. Overpopulation may lead to many societal problems as well as to changes in the habitat. Birth control measures are thus needed to control this growth. However, for the last 50-60 years, there have not been any improvements in the field of contraception. Nevertheless, the immunocontraceptive vaccine is an emerging field, and it might be the only replacement for the existing mode of contraception for the next millennium. Sexually transmitted infections (STIs) are frequent, and their transmission rate increases yearly. As antibiotics are the prevailing treatment for this kind of infections, resistance in humans has increased; therefore, having effective antibiotic treatments for STIs is now a concern. Vaccines against STIs are now needed. It is thought that the improvements in the fields of proteomics, immunomics, metabolomics, and other omics will help in the successful development of vaccines. Objective: To collect and review the literature about recent advancements in immunocontraception and vaccines against sexually transmitted diseases/infections. Methods: Reliable scientific databases, such as PubMed Central, PubMed, Scopus, Science Direct, and Goggle Scholar, were consulted. Publications bearing important information on targeted antigens/immunogens for contraceptive vaccine design and advancements in vaccine development for STIs were gathered and tabulated, and details were analyzed as per the theme of each study. Results: Important antigens that have a specific role in fertility have been studied extensively for their contraceptive nature. Additionally, the advancements in the screening for the best antigens, according to their antigenic nature and how they elicit immune responses for an extended period were also studied. Herd immunity for STIs and advancements in the development of vaccines for syphilis, gonorrhea, and herpes simplex virus were also studied and tabulated in this review. An extensive knowledge on STIs vaccines was gained. Conclusion: This extensive review is aimed to provide insights for active researchers in vaccinology, immunology, and reproductive biology. Advancements in the development of vaccines for different STIs can be gathered as a wholesome report.

Nanotechnology-assisted production of value-added biopotent energy-yielding products from lignocellulosic biomass refinery - A review.

The need to develop sustainable alternatives for pretreatment and hydrolysis of lignocellulosic biomass (LCB) is a massive concern in the industrial sector today. Breaking down of LCB yields sugars and fuel in the bulk scale. If explored under nanotechnology, LCB can be refined to yield high-performance fuel sources. The toxicity and cost of conventional methods can be reduced by applying nanoparticles (NPs) in refining LCB. Immobilization of enzymes onto NPs or used in conjugation with nanomaterials would instill specific and eco-friendly options for hydrolyzing LCB. Nanomaterials increase the proficiency, reusability, and stability of enzymes. Notably, magnetic NPs have bagged their place in the downstream processing of LCB effluents due to their efficient separation and cost-effectiveness. The current review highlights the role of nanotechnology and its particles in refining LCB into various commercial precursors and value-added products. The relationship between nanotechnology and LCB refinery is portrayed effectively in the present study.

Role of nanomaterials in deactivating multiple drug resistance efflux pumps - A review.

The changes in lifestyle and living conditions have affected not only humans but also microorganisms. As man invents new drugs and therapies, pathogens alter themselves to survive and thrive. Multiple drug resistance (MDR) is the talk of the town for decades now. Many generations of medications have been termed useless as MDR rises among the infectious population. The surge in nanotechnology has brought a new hope in reducing this aspect of resistance in pathogens. It has been observed in several laboratory-based studies that the use of nanoparticles had a synergistic effect on the antibiotic being administered to the pathogen; several resistant strains scummed to the stress created by the nanoparticles and became susceptible to the drug. The major cause of resistance to date is the efflux system, which makes the latest generation of antibiotics ineffective without reaching the target site. If species-specific nanomaterials are used to control the activity of efflux pumps, it could revolutionize the field of medicine and make the previous generation resistant medications active once again. Therefore, the current study was devised to assess and review nanoparticles' role on efflux systems and discuss how specialized particles can be designed towards an infectious host's particular drug ejection systems.

Characterization of human prostasomes protein Clusterin (macromolecule) - a novel biomarker for male infertility diagnosis and prognosis.

Prostasomes, a secretory particle from prostate gland in human seminal fluid plays a role in enhancing the fertility and its absence or less presence will lead to male infertility. Very few fertility associated proteins were detected in prostasomes. In order to isolate the prostasomes from ejaculated semen, the characterization was done using biochemical, molecular and in silico methods. The objective of current work on prostasomes is to identify a novel protein biomarker for the diagnosis and prognosis of male infertility. Semen samples were collected, primary semen analysis was done and prostasomes were isolated from ejaculated semen of fertile and infertile categories. Biochemical parameters like protein content, total antioxidant capacity, cholesterol content were evaluated in prostasomes. The critical expressed protein was identified by using SDS page and MALDI TOF techniques. Isolated particle from the semen samples was confirmed as prostasomes from SEM analysis and particle size analyzer. In MALDI results, the maximum hit was obtained against Clusterin that been reported to be involved in capacitation and motility of sperm. Structural modeling and molecular dynamics were carried out on Clusterin and elucidate the structural insights in the dynamic system. Overall, our study reported the novel biomarker Clusterin in prostasomes for diagnosis and prognosis of male infertility.Communicated by Ramaswamy H. Sarma.

Identification and Structure Prediction of Human Septin-4 as a Biomarker for Diagnosis of Asthenozoospermic Infertile Patients-Critical Finding Toward Personalized Medicine.

Semen parameters are been found as a key factor to evaluate the count and morphology in the given semen sample. The deep knowledge of male infertility will unravel with semen parameters correlated with molecular and biochemical parameters. The current research study is to identify the motility associated protein and its structure through the in-silico approach. Semen samples were collected and initial analysis including semen parameters was analyzed by using the World Health Organization protocol. Semen biochemical parameters, namely, seminal plasma protein concentration, fructose content, and glucosidase content were calculated and evaluated for correlation. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) were carried out for identification of Septin-4 presence in the semen sample. Mascot search was done for protein conformation and in-silico characterization of Septin-4 by structural modeling in Iterative Threading Assembly Refinement (I-TASSER). Twenty-five nanoseconds molecular dynamics (MD) simulations results showed the stable nature of Septin-4 in the dynamic system. Overall, our results showed the presence of motility-associated protein in normospermia and control samples and not in the case of asthenospermia and oligoasthenospermia. Molecular techniques characterized the presence of Septin-4 and as a novel biomarker for infertility diagnosis.

Efficacy of encapsulated biogenic silver nanoparticles and its disease resistance against Vibrio harveyi through oral administration in Macrobrachium rosenbergii.

Biological synthesis of silver nanoparticles (AgNPs) by Cheatomorpha antennia and its in vitro and in vivo antibacterial activity against Vibrio harveyi in Macrobrachium rosenbergii was demonstrated in the study. In vitro growth curve analysis, cell viability and bacterial inhibitory assays were performed to test the efficacy of synthesised AgNPs against bacteria. Sodium caseinate was used as an encapsulating agent to deliver the antibacterial drugs and the commercial process of microencapsulation comprises the antibacterial bioelements for oral administration to improve the disease resistance of AgNPs against V. harveyi due to the eco-friendly for non-toxic behaviour of nanoparticle and their treatment. Characterisation of antibacterial silver was performed by UV spectroscopy, X-ray diffraction, Fourier Transform Infrared spectroscopy and Scanning Electron Microscopy. The peak at 420 nm showed the presence of nanoparticles in the solution and the crystal nature of the particle was identified by the XRD. FTIR characterised the functional harveyi biomolecules and further SEM confirmed the size of the nanoparticles around 24 ± 2.4 nm. Experimental pathogenicity of V. harveyi showed 100% mortality at the 120th hour. Treatment of encapsulated AgNPs was administered orally for the relative percentage of survival which acquired almost 90% of survival till 30 days of exposure. In conclusion, the microencapsulation of AgNPs in the biopolymer matrices promotes the health, growth responses, immunity and disease resistance of encapsulated AgNPs with an improved relative percentage of survival.

Alpha-Synuclein Aggregation in Parkinson's Disease.

Parkinson's disease (PD), a neurodegenerative disorder characterized by distinct aging-independent loss of dopaminergic neurons in substantia nigra pars compacta (SNpc) region urging toward neuronal loss. Over the decade, various key findings from clinical perspective to molecular pathogenesis have aided in understanding the genetics with assorted genes related with PD. Subsequently, several pathways have been incriminated in the pathogenesis of PD, involving mitochondrial dysfunction, protein aggregation, and misfolding. On the other hand, the sporadic form of PD cases is found with no genetic linkage, which still remain an unanswered question? The exertion in ascertaining vulnerability factors in PD considering the genetic factors are to be further dissevered in the forthcoming decades with advancement in research studies. One of the major proponents behind the prognosis of PD is the pathogenic transmutation of aberrant alpha-synuclein protein into amyloid fibrillar structures, which actuates neurodegeneration. Alpha-synuclein, transcribed by SNCA gene is a neuroprotein found predominantly in brain. It is implicated in the modulation of synaptic vesicle transport and eventual release of neurotransmitters. Due to genetic mutations and other elusive factors, the alpha-synuclein misfolds into its amyloid form. Therefore, this review aims in briefing the molecular understanding of the alpha-synuclein associated with PD.

Ecotoxicological assessment and dermal layer interactions of nanoparticle and its routes of penetrations.

Our review focused on nanomaterials-based toxicity evaluation and its exposure to the human and aquatic animals when it was leached and contaminated in the environment. Ecotoxicological assessment and its mechanism mainly affect the skin covering layers and its preventive barriers that protect the foreign particles' skin. Nanoscale materials are essential in the medical field, especially in biomedical and commercial applications such as nanomedicine and drug delivery, mainly in therapeutic treatments. However, various commercial formulations of pharmaceutical drugs are manufactured through a series of clinical trials. The role of such drugs and their metabolites has not met the requirement of an individual's need at the early stage of the treatments except few drugs and medicines with minimal or no side effects. Therefore, biology and medicines are taken up the advantages of nano scaled drugs and formulations for the treatment of various diseases. The present study identifies and analyses the different nanoparticles and their chemical components on the skin and their effects due to penetration. There are advantageous factors available to facilitate positive and negative contact between dermal layers. It creates a new agenda for an established application that is mainly based on skin diseases.

Seminal exosomes - An important biological marker for various disorders and syndrome in human reproduction.

BACKGROUND: Exosomes are nano-sized membrane vesicles, secreted by different types of cells into the body's biological fluids. They are found in abundance in semen as compared to other fluids. Exosomes contain a cargo of lipid molecules, proteins, phospholipids, cholesterol, mRNAs, and miRNAs. Each molecule of seminal exosomes (SE) has a potential role in male reproduction for childbirth. Many potential candidates are available within the seminal exosomes that can be used as diagnostic markers for various diseases or syndromes associated with male reproduction. Also these seminal exospmes play a major role in female reproductive tract for effective fertilization. AIM: The aim of this review is to focus on the advancement of human seminal exosomal research and its various properties. METHODS: We used many databases like Scopus, Google scholar, NCBI-NLM and other sources to filter the articles of interest published in exosomes. We used phrases like "Exosomes in human semen", "Composition of exosomes in human semen" and other relevant words to filter the best articles. RESULTS: Seminal exosomes play a major role in sperm functions like cell-to-cell communication, motility of the sperm cells, maintaining survival capacity for the sperm in the female reproductive tract and spermatogenesis. Also, seminal exosomes are used as a carrier for many regulatory elements using small RNA molecules. miRNAs of the seminal exosomes can be used as a diagnostic marker for prostate cancer instead of prostate specific antigen (PSA). Epididymosomes can be used as a biomarker for reproductive diseases and male infertility. CONCLUSION: Seminal exosomes could be used as biological markers for various reproductive disorders, male infertility diagnosis, and it can be used in anti-retroviral research for the identification of novel therapeutics for HIV-1 infection and transmission.

Decoding Conformational Imprint of Convoluted Molecular Interactions Between Prenylflavonoids and Aggregated Amyloid-Beta42 Peptide Causing Alzheimer's Disease.

Protein misfolding occurs due to the loss of native protein structure and adopts an abnormal structure, wherein the misfolded proteins accumulate and form aggregates, which result in the formation of amyloid fibrils that are associated with neurodegenerative diseases. Amyloid beta (Aß42) aggregation or amyloidosis is contemplated as a unique hallmark characteristic of Alzheimer's disease (AD). Due to aberrant accrual and aggregation of Aß42 in extracellular space, the formation of senile plaques is found in AD patients. These senile plaques occur usually in the cognitive and memory region of the brain, enfeebles neurodegeneration, hinders the signaling between synapse, and disrupts neuronal functioning. In recent years, herbal compounds are identified and characterized for their potential as Aß42 inhibitors. Thus, understanding their structure and molecular mechanics can provide an incredible finding in AD therapeutics. To describe the structure-based molecular studies in the rational designing of drugs against amyloid fibrils, we examined various herbal compounds that belong to prenylflavonoids. The present study characterizes the trends we identified at molecular docking studies and dynamics simulation where we observed stronger binding orientation of bavachalcone, bavachin, and neobavaisoflavone with the amyloid-beta (Aß42) fibril structure. Hence, we could postulate that these herbal compounds could be potential inhibitors of Aß42 fibrils; these anti-aggregation agents need to be considered in treating AD.

Identification of curcumin derivatives as human LMTK3 inhibitors for breast cancer: a docking, dynamics, and MM/PBSA approach.

Human lemur tyrosine kinase-3 (LMTK3) is primarily involved in regulation of estrogen receptor-α (ERα) by phosphorylation activity. LMTK3 acts as key biomarker for ERα positive breast cancer and identified as novel drug target for breast cancer. Due to the absence of experimental reports, the computational approach has been followed to screen LMTK3 inhibitors from natural product curcumin derivatives based on rational inhibitor design. The initial virtual screening and re-docking resulted in identification of top three leads with favorable binding energy and strong interactions in critical residues of ATP-binding cavity. ADME prediction confirmed the pharmacological activity of the leads with various properties. The stability and binding affinity of leads were well refined in dynamic system from 25 ns MD simulations. The behavior of protein motion towards closure of ATP-binding cavity was evaluated based on eigenvectors by PCA. In addition, MM/PBSA calculations also confirmed the relative binding free energy of LMTK3-lead complexes in favor of the effective binding. From our study, novel LMTK3 inhibitors tetrahydrocurcumin, curcumin 4,4'-diacetate, and demethoxycurcumin have been proposed with inhibition mechanism. Further experimental evaluation on reported lead candidates might prove its role in breast cancer therapeutics.

Designing and optimization of novel human LMTK3 inhibitors against breast cancer - a computational approach.

Estrogen receptor-α (ERα) is expressed more in patients with breast cancer and its level correlated with endocrine resistance. LMTK3 is reported as breast cancer target with regulation of estrogen receptor-α (ERα) through phosphorylation. In this computational study, structure-based inhibitor screening was performed on human LMTK3 using ZINC database. ATP-binding cavity with critical residues involved in the LMTK3 phosphorylation was used as target site for the screening. From the large ligand library, the best compounds were screen with three-phase virtual screening methods in Dockblaster, AutoDock Vina and AutoDock, respectively. The evaluation of ligands was carried out by binding energy and weak interactions, such as hydrogen bond interactions and hydrophobic contacts, in the target site that favors LMTK3 inhibition. Top compounds were found to be more effective in druglikeness activity by ADME prediction. The stability and binding affinity of ligand complexes were optimized by trajectory analysis such as RMSD, Rg, SASA and interhydrogen bonds from molecular dynamics simulations. The behavior of protein motion after ligand binding was illustrated by eigenvectors from principal component analysis (PCA). In addition, binding free energy of the LMTK3-ligand complexes were calculated by MM/PBSA methods and results supported the strong binding in dynamic system. Thus, the computational studies illustrated the structural insights on LMTK3 inhibition mechanism by ligands ZINC04670539, ZINC05607079 and ZINC04344028, also proposed as potent lead candidates. Our findings step towards developing novel LMTK3 inhibitors and identified lead candidates can be future breast cancer drugs with further experimental studies.

Quercetin-3-O-rhamnoside from Euphorbia hirta protects against snake Venom induced toxicity.

BACKGROUND: The plant Euphorbia hirta is widely used against snake envenomations in rural areas and it was proved to be effective in animal models. Therefore, the scientific validation of its phytoconstituents for their antiophidian activity is aimed in the present study. METHODS: E. hirta extract was subjected to bioactivity guided fractionation and the fractions that inhibited different enzyme activities of Naja naja venom in vitro was structurally characterized using UV, FT-IR, LC-MS and NMR spectroscopy. Edema, hemorrhage and lethality inhibition activity of the compound were studied in mice model. In addition, molecular docking and molecular dynamic simulations were also performed in silico. RESULTS: The bioactive fraction was identified as Quercetin-3-O-α-rhamnoside (QR, 448.38 Da). In vitro experiments indicated that protease, phospholipase-A(2), hemolytic activity and hemorrhage inducing activity of the venom were inhibited completely at a ratio of 1:20 (venom: QR) w/w. At the same concentration, the edema ratio was drastically reduced from 187% to 107%. Significant inhibition (93%) of hyaluronidase activity was also observed at a slightly higher concentration of QR (1:50). Further, in in vivo analysis, QR significantly prolonged the survival time of mice injected with snake venom. CONCLUSION: For the first time Quercetin-3-O-α-rhamnoside, isolated from E. hirta, has been shown to exhibit anti-snake venom activity against Naja naja venom induced toxicity. GENERAL SIGNIFICANCE: Exploring such multifunctional lead molecules with anti-venom activity would help in developing complementary medicine for snakebite treatments especially in rural areas where anti-snake venom is not readily available.

Induction of deletion mutation on ompR gene of Salmonella enterica serovar Typhi isolates from asymptomatic typhoid carriers to evolve attenuated strains for vaccine development.

OBJECTIVE: To develop attenuated strains of Salmonella enterica serovar Typhi (S. typhi) for the candidate vaccine by osmolar stress. METHODS: S. typhi SS3 and SS5 strains were isolated from asymptomatic typhoid carriers in Namakkal, Tamil Nadu, India. Both strains were grown in LB (Luria Bertani) medium supplemented with various concentration of NaCl (0.1-0.7M) respectively. The effect of osmolar stress was determined at molecular level by PCR using MGR 06 and MGR 07 primers corresponding to ompR with chromosomal DNA of S. typhi SS3 and SS5 strains. Attenuation by osmolar stress results in deletion mutation of the S. typhi strains was determined by agglutination assays, precipitation method, SDS PAGE analysis and by animal models. RESULTS: The 799 bp amplified ompR gene product from wild type S. typhi SS3 and SS5 illustrate the presence of virulent gene. Interestingly, there was only a 282 bp amplified product from S. typhi SS3 and SS5 grown in the presence of 0.5, 0.6 and 0.7 M NaCl. This illustrates the occurrence of deletion mutation in ompR gene at high concentration of NaCl. Furthermore, both the wild-type and mutant S. typhi outer membrane SDS-PAGE profile reveals the differences in the expression of ompF, ompC and ompA proteins. In mice, wild type and mutant strains lethal dose (LD50) were determined. The mice died within 72 h when both the wild type strains were injected intraperitoneally with 3 log CFU.mL(-1). When the mice were injected with the mutants in same dosage, no clinical symptoms were observed; whereas the serum antibody titre was elicited within two weeks indicated that the mutants have the ability to induce protective humoral immune response. These results suggest that S. typhi SS3 and SS5 may be used as good candidate strains for the development of live attenuated vaccine against salmonellosis. CONCLUSIONS: This study demonstrates that the S. typhi strains were attenuated and could be good vaccine candidates in future.

Whey powder: a potential anti-diarrheal agent through its biofilm formation.

Whey, the natural product resulting from coagulation of milk is reported to have diverse pharmaceutical credentials. In the present investigation the anti-diarrhoeal activity of the whey powder was investigated. The Whey powder which was prepared using rennet powder and lactic acid, was studied against Magnesium sulphate-induced Diarrhea in Swiss Albino mice. Castor oil-induced enteropooling studies and in vitro biofilm-forming potentials of the whey powder were also carried out, as this is believed to contribute to the anti-diarrhoeal activities of the preparation. Anti-diarrhoeal activity was more pronounced in mice which received 250 mg kg b.wt. of whey powder when compared to those which received 500 mg kg(-1) b.wt. The percentage inhibition of total number of feces in the 250 mg kg(-1) b.wt. drug-treated group was 56.14%,whereas the animals which received 500 mg kg(-1) b.wt. of whey powder showed 37.18% inhibition. The loperamide treated animal group showed 63.81% inhibition. In castor oil induced enteropooling, the percentage inhibition of intestinal content in the 250 mg kg(-1) b.wt. drug-treated group was 61.42% against atropine-treated animal group that showed 26.24% inhibition. The whey powder also exhibited strong biofilm forming capacity with increase in concentration. The anti-diarrhoeal activity of whey preparation established herein is believed to be owing to certain active principles present in it or due to biofilm-forming capacity, which inhibits the attachment of mediators of diarrhoea to mucosal walls of the GI tract or due to interaction of diarrhoea inducing chemicals with whey peptides, which needs further investigation.

Structural modeling and molecular dynamics studies on the human LMTK3 domain and the mechanism of ATP binding.

Estrogen positive breast cancer is a dreadful disease in women worldwide. The human estrogen receptor-α (ERα) pathway plays a critical role in estrogenic signaling and targeting ERα in breast cancer treatment. The key role of Lemur tyrosine kinase-3 (LMTK3) in regulation of ERα has been identified and it is found to be a novel therapeutic target for breast cancer. With lack of structural studies on LMTK3, the breast cancer therapeutics research remains elusive. In this computational study, we performed structural studies on LMTK3 by structural modeling and molecular dynamics (MD) simulations of the apo state and the ATP bound state. The structure of the LMTK3 domain was developed by using I-TASSER server and validated by quality index and Ramachandran plot. MD simulation analysis explained the structural behavior of the LMTK3 domain in the dynamic system and the apo state showed defined protein folding with stable conformation. The mechanism of ATP binding was studied using molecular docking, resulting in the identification of critical residues and the ATP binding cavity. Furthermore, MD simulation of the LMTK3-ATP complex was performed and the trajectory analyses confirmed the stability and effective binding of ATP in the dynamic system. Overall, our computational reports provide more information on the structure-function relationship of LMTK3 with ATP. The critical residues Tyr185 and Asp284 found in the ATP binding cavity may be useful in designing potential inhibitors on human LMTK3.

From natural products to drugs for epimutation computer-aided drug design.

The epimutational event, i.e., ectopic methylation in tumor suppressor genes, can lead to gene silencing, thus promoting prognosis of cancer. The progression of DNA methylation is a cycle of demethylation, de novo methylation, and maintenance methylation. The enzyme responsible for maintenance of methylation status is DNA methyltransferase 1 (DNMT1), the continuous activity of which is required to maintain the pattern of epimutation; thus, its inhibition is a promising strategy for the treatment of cancer. To the best of our knowledge, this study is the first to focus on the recently developed crystal structure of the catalytic site of DNMT1. Here in this study, we have used the crystal structure for the development of non-nucleoside DNMT1 inhibitors using virtual screening (VS), absorption, distribution, metabolism, elimination/toxicology analysis, and molecular docking studies. In this study, VS was carried out on 48,531 natural products to create a subset of lead-like natural products. Three of them were found to form hydrogen bonds with the catalytic site of the DNMT1 (Cys 1226). Thus, this study adumbrates potential lead compounds for treatment of epimutation.

The E3 ubiquitin ligase Itch regulates tumor suppressor protein RASSF5/NORE1 stability in an acetylation-dependent manner.

Ras association (RalGDS/AF-6) domain family member RASSF5 is a non-enzymatic RAS effector super family protein, known to be involved in cell growth regulation. Expression of RASSF5 is found to be extinguished by promoter hypermethylation in different human cancers, and its ectopic expression suppresses cell proliferation and tumorigenicity. Interestingly, this role in tumorigenesis has been confounded by the fact that regulation at molecular level remains unclear and many transformed cells actually display elevated RASSF5 expression. Here, we demonstrate that E3 ubiquitin ligase Itch is a unique binding partner of RASSF5. Itch can interact with PPxY motif in RASSF5 both in vivo and in vitro through its WW domains. Importantly, the overexpression of Itch induces RASSF5 degradation by poly-ubiquitination via 26S proteasome pathway. In addition, our results indicate that the elevated levels of RASSF5 found in tumor cells due to acetylation, which restricts its binding to Itch and results in a more stable inert protein. Inhibition of RASSF5 acetylation permits its interaction with Itch and provokes proteasomal degradation. These data suggest that apart from promoter methylation, hyperacetylation could also be downregulating RASSF5 function in different human cancer. Finally, results from functional assays suggest that the overexpression of wild type, not the ligase activity defective Itch negatively regulate RASSF5-mediated G1 phase transition of cell cycle as well as apoptosis, suggesting that Itch alone is sufficient to alter RASSF5 function. Collectively, the present investigation identifies a HECT class E3 ubiquitin ligase Itch as a unique negative regulator of RASSF5, and suggests the possibility that acetylation as a potential therapeutic target for human cancer.