Enterobacter cloacae is mainly responsible for sepsis, urethritis, and respiratory tract infections. These bacteria may affect the transcription of the host and particularly their immune system by producing changes in their epigenetics. In the present study, four proteins of Enterobacter cloacae were used to predict the epitopes for the construction of an mRNA vaccine against Enterobacter cloacae infections. In order to generate cellular and humoral responses, various immunoinformatic-based approaches were used for developing the vaccine. The molecular docking analysis was performed for predicting the interaction among the chosen epitopes and corresponding MHC alleles. The vaccine was developed by combining epitopes (thirty-three total), which include the adjuvant Toll-like receptor-4 (TLR4). The constructed vaccine was analyzed and predicted to cover 99.2% of the global population. Additionally, in silico immunological modeling of the vaccination was also carried out. When it enters the cytoplasm of the human (host), the codon is optimized to generate the translated mRNA efficiently. Moreover, the peptide structures were analyzed and docked with TLR-3 and TLR-4. A dynamic simulation predicted the stability of the binding complex. The assumed construct was considered to be a potential candidate for a vaccine against Enterobacter cloacae infections. Hence, the proposed construct is suitable for in vitro analyses to validate its effectiveness.

Serratia marcescens, a Gram-negative bacterium, is one of the known disease-causing pathogens. It is resistant to ampicillin, macrolides, cephalosporins, cefotaxime, and ceftazidime. The only antibiotic that has been proven to be effective against S. marcescens is gentamicin. By causing epigenetic alterations, bacteria can also become resistant to all antibiotics. Many epigenetically related proteins were studied, and four proteins were selected in this regard for epitope evaluation and their subsequent use in the development of a messenger ribonucleic acid (mRNA) vaccine. A series of immune-informatics tools used to build this mRNA vaccine elicited cellular and humoral immunity. Molecular docking between epitopes and alleles of the major histocompatibility complex (MHC) was performed. The vaccine was developed using 37 epitopes, an adjuvant that is a TLR-4 agonist known as resuscitation-promoting factor E (RpfE), subcellular trafficking structures, secretion boosters, and linkers. This proposed architecture was found to cover 99.6% of the population during testing. During testing, it was proven that it was both effective and safe. To confirm our idea, we performed an in silico immunological simulation of vaccination. The codon was also optimized to ensure that the mRNA reached the cytoplasm of a human host and underwent efficient translation. TLR-4 and TLR-3 were also docked against the secondary and tertiary structures of the vaccine peptide. Furthermore, the vaccine's stability was confirmed by molecular dynamics simulation. In summary, this vaccine construct can be a potential candidate against S. marcescens and is suitable for in vitro analyses to validate its effectiveness.

Hypothetical proteins (HPs) are non-predicted sequences that are identified only by open reading frames in sequenced genomes but their protein products remain uncharacterized by any experimental means. The genome of every species consists of HPs that are involved in various cellular processes and signaling pathways. Annotation of HPs is important as they play a key role in disease mechanisms, drug designing, vaccine production, antibiotic production, and host adaptation. In the case of bacteria, 25-50% of the genome comprises of HPs, which are involved in metabolic pathways and pathogenesis. The characterization of bacterial HPs helps to identify virulent proteins that are involved in pathogenesis. This can be done using in-silico studies, which provide sequence analogs, physiochemical properties, cellular or subcellular localization, structure and function validation, and protein-protein interactions. The most diverse types of virulent proteins are exotoxins, endotoxins, and adherent virulent factors that are encoded by virulent genes present on the chromosomal DNA of the bacteria. This review evaluates virulent HPs of pathogenic bacteria, such as Staphylococcus aureus, Chlamydia trachomatis, Fusobacterium nucleatum, and Yersinia pestis. The potential of these HPs as a drug target in bacteria-caused infectious diseases along with the mode of action and treatment approaches have been discussed.

In the modern era, nanotechnology is one of the rapidly growing fields in many biomedical applications especially in delivering therapeutics for the curation of tumor cells. Cancer is a lethal disease that grows silently in the human body, caused by mutations in the genetic material of cells. The presence of a high number of physiologically active chemicals on the cancer cell surface does not particularly lead to tumor cell targeting. The lack of effective targeting of tumor cells by conventionally administered chemotherapeutic drugs demands the use of nanomedicine. Conventional medicines have immense cytotoxic effects, i.e., they do not kill tumor cells only but also destroy normal body cells. Nanoparticles (NPs) have an advantage over conventional therapeutic methods due to their small size, large surface area, and especially for targeting tumor cells without harming normal body cells. Based on durability and molecular weight, NPs are classified as organic and inorganic nanoparticles. Common organic and inorganic nanoparticles are highly effective against cancer curation and have high efficacy values along with their mechanism of action.

Epidermolysis Bullosa is a rare genetic disorder that causes skin fragility, trauma induced dissociations of the skin, and painful wound growth. More than 20 types of genes are involved in causing EB as it is a polygenic disease and each gene is involved in a different subtype of EB. Dystrophic Epidermolysis Bullosa (DEB) is one of the subtypes of EB caused by mutations in the COL7A1 (Collagen Type VII Alpha 1 Chain) gene and it affects people from all racial backgrounds. No drug is available for DEB in the market yet. So, it is the need of the hour to come up with a potential inhibitor that could inhibit the faulty protein of COL7A1 gene. Different exons of COL7A1 have been analyzed and exon no 70 to 75 has been selected which were important for mutational point of view. The mutations in it have been identified and verified using various In-silico tools. The 3D structure of the protein has been retrieved using specific exons which were edited and mutations were introduced in it and it was further checked to analyze its stability, toxicity and solubility of the protein. The inhibitors of COL7A1 have been formed using CAAD techniques (pharmacophore modeling) and the best inhibitor of COL7A1 has been further checked to determine its drug-likeness, solubility, its toxicity, and various physiochemical properties. The constructed inhibitor was found to have the best docking results and found to have good ADMET properties. The developed inhibitor construct showing promising results In-silico and it will also show good results if it would be tested in-vitro and in-vivo. Thus, it would be a breakthrough to treat DEB using this inhibitor if this inhibitor is constructed and further tested in-vitro.