Current Protein and Peptide Science - Online First

The pathogenesis of age-related hearing loss (ARHL), especially the role of long non-coding RNAs (lncRNAs) and their encoded peptides, remains incompletely understood. This study aimed to characterize expression changes in lncRNAs and peptides in the cochleae of ARHL mice and explore the potential functions of lncRNA-encoded peptides via multi-omics analysis.

C57BL/6J mice were used to establish the ARHL model. The molecular expression profiles of cochlear tissues from normal and ARHL mice were synthesized by lncRNA sequencing, peptidomics, and bioinformatics.

Compared with the control group, a total of 789 differentially expressed lncRNAs and 466 differentially expressed peptides were identified in the ARHL group. Functional enrichment analysis revealed their association with key pathways, including ion transport, calcium signaling, the TCA cycle, and cytoskeleton regulation, indicating broad molecular dysregulation in the aging cochlea. Notably, 64 differentially expressed lncRNAs showed high translational potential, yielding 107 novel lncRNA-encoded peptides. These were mainly short peptides, some with stabilizing hydrophobic properties suited for membrane interactions, and enriched in domains like Pkinase and C2, suggesting involvement in signal transduction.

These results emphasized that lncRNA-encoded peptides were novel regulators of ARHL, potentially regulating calcium homeostasis and mitochondrial function. The overlap of pathways such as the cytoskeleton and fatty acid metabolism indicated that the lncRNA-peptide axis drove auditory decline, providing institutional insights into the epigenetic basis of ARHL.

Our findings suggest that lncRNA-encoded peptides are a novel class of regulatory molecules involved in the complex pathogenesis of ARHL, highlighting them as promising targets for future therapeutic strategies.

Parkinson's disease (PD), a condition that involves neural degeneration, develops due to dopaminergic neuronal death in the substantia nigra pars compacta, resulting in reduced striatal dopamine levels. This shortage causes problems with movement and thinking. The neurodefensive response of GLP-1 is especially important in PD. Assessments have demonstrated the neuroprotective advantages of activating GLP-1 receptors in distinct models of PD, resulting in enhancements in motor as well as non-motor behaviour. These characteristics suggest that GLP-1 signalling could be a promising target for PD treatment. Moreover, plumbagin is the primary active component of Plumbago zeylanica L., a medicinal herb that is clinically used in China. Also, plumbagin is reported to have significant neuroprotective efficacy.

In this study, male rats received rotenone (1.5 mg/kg; subcutaneously), followed by plumbagin (20 mg/kg; p.o.). The rats’ motor abilities were assessed using the actophotometer, beam walk, rotarod, gait analysis, open field, grip strength, as well as bar catalepsy evaluation. In addition, the levels of dopamine, RAGE, and GLP-1 were measured.

Plumbagin improved movement issues caused by rotenone, boosted dopamine and GLP-1 levels, as well as lowered RAGE levels in the brains of rats.

The study highlights plumbagin’s potential in treating PD by improving motor function, increasing dopamine and GLP-1 levels, and reducing RAGE levels in a rotenone-induced rat model. These findings suggest that plumbagin may offer neuroprotective effects through GLP-1 pathway activation, making it a promising candidate for future PD therapies.

These outcomes imply that agents that activate GLP-1, such as plumbagin, present a promising strategy for creating treatments to safeguard against rotenone-induced motor disorders.

Microbial enzymes, especially bacterial alkaline proteases, are essential to many industrial processes, including the manufacturing of detergents, food processing, bioremediation, medicines, and tanneries. Because of its possible industrial benefits, this study focuses on the purification and characterisation of a halophilic alkaline protease generated by Bacillus sp. strain SPII-4.

The bacteria SPII-4's 16S rRNA gene was sequenced and subjected to phylogenetic analysis. Casein was used as a substrate to measure the extracellular crude enzyme's proteolytic activity. Temperature, pH, salinity, metal ions, and chemical solvents were all used to assess enzymatic activity. Every experiment was run in triplicate, and Student's t-tests with unequal variances in Microsoft Excel were used to assess statistical significance.

The 16S rRNA sequencing matched Bacillus sp. strain 2S4 with 100% identity and 99% coverage. The protease was most active at 40°C, in the alkaline pH range of 9-11, and at concentrations of up to 5% NaCl. The enzyme had the maximum activity (14.64 U/mg) among the metal ions examined when BaCl2 was present. Additionally, it maintained its activity in the presence of the surfactant Triton-X and in a variety of chemical solvents. The observed differences were statistically significant (p < 0.001).

The Bacillus SPII-4 protease showed exceptional stability and activity in the presence of surfactants and solvents, as well as in extremely high and low salinity and alkalinity conditions. These characteristics point to the protease's potential for widespread industrial use and are in line with research on related halophilic bacterial enzymes. To maximize its commercial usage, more purification and scale-up research are necessary.

Bacillus sp. SPII-4's halo-alkaline protease exhibits considerable industrial promise because of its stability in conditions that are high in salt, alkalinity, and solvents. These qualities make it a viable option for use in the food, detergent, and pharmaceutical sectors as well as in bioremediation.

Periodontitis results in progressive loss of gingival tissue and periodontal ligament, eventually resulting in tooth instability. As regenerating degraded periodontal tissue is not possible without intervention, therefore, a tissue-engineered substitute is a good option. Bone regeneration strategies often rely on either biochemical stimulation or engineered scaffolds, but rarely in a coordinated manner. Arginine-Glycine-Aspartic acid (RGD) hydrogel provides a unique combination of biocompatibility and biodegradability, making it an attractive scaffold for tissue engineering. The study aims to investigate the effect of combining Wnt pathway activation with Arginine-Glycine-Aspartic acid (RGD) hydrogel (a three-dimensional environment, 3D) to enhance the osteogenic differentiation of mesenchymal stem cells (MSCs) derived from periodontal ligament tissue.

The cells were isolated from the root of the extracted tooth. They were grown in an osteogenic medium with and without a Wnt activator in two-dimensional (2D) and RGD hydrogel-based 3D environments to expand in vitro. Osteogenic gene expression was evaluated by qPCR in 2D and 3D cultures. Mesenchymal stem cells isolated from periodontal ligament tissue showed osteogenic differentiation when cultured in a differential medium with or without the Wnt signaling activator, CHIR99021 (a GSK3β inhibitor).

The data of our study revealed that osteogenic genes were expressed in both 2D- and 3D-cultured cells. However, higher expression of osteogenic genes was found in Wnt signaling-activated cells. Furthermore, the RGD hydrogel provided better differentiation efficacy and a significant increase (p < 0.001) in terms of Wnt-activated differentiation.

The RGD hydrogel-Wnt activation model described in this study holds strong potential for translation into preclinical bone regeneration strategies. By enhancing osteogenic differentiation through a synergistic interaction between the Wnt signaling pathway and the 3D peptide hydrogel matrix, this platform offers a promising approach to early-stage testing of bone regeneration therapies.

Hence, the Arg-Gly-Asp (RGD) hydrogel-based 3D microenvironment along with a Wnt signaling activator provides superior efficacy in differentiation since it allows cell encapsulation and an environment that closely simulates native tissues. Therefore, these findings highlight the synergistic effect of biochemical and biophysical cues in directing stem cell fate and offer a promising strategy for advancing stem cell-based bone tissue engineering.

One of the earliest illnesses that has been identified is tuberculosis (TB). The largest challenge in managing tuberculosis today is the growing number of individuals infected with TB bacilli, particularly those that are Extensively drug resistant-tuberculosis (XDR-TB) and Multidrug-resistant tuberculosis (MDR-TB). However, by figuring out the resistance's molecular mechanism, Advanced molecular methods may be used to rapidly determine therapy plans. Combining Delamanid (DLM) with Bedaquiline (BDQ), one of the recently authorized medications, indicates that the therapy is effective.

We aim to investigate efflux-mediated resistance mechanisms in M. Tuberculosis by using quantitative real-time PCR to assess the expression level of mmpS5 and mmpL5.

The median (M) and interquartile range (Iqr) of mmpL5 and mmpS5 expression varied from 5.65 to 9.01 and 7.95 to 10.74, respectively, when resistant strains were compared with sensitive ones. M and Iqr of mmpL5 and mmpS5 expression, however, ranged from 0.08–3.04 and 0.05–1.61 for sensitive strains, correspondingly.

Our findings have implications for the development of fast genotypic drug susceptibility testing (DST). Quantitative real-time PCR to measure the expression level of mmpS5 and mmpL5 of baseline and post-baseline isolates is important to track the development of BDQ and DLM resistance.

Thus, when developing anti-tuberculosis drugs, mycobacterial MmpS5-MmpL5 transporters should be taken into consideration early on, as they are an MDR-efflux system.

One to two percent of women worldwide experience recurrent pregnancy loss (RPL), defined as the loss of two or more consecutive pregnancies before 20 weeks of gestation. Genetic factors, including variations in the FOXP3 gene, have been implicated in the unexplained etiology of RPL. This study aimed to identify and characterize novel genetic variants in exons 2 and 7 of the FOXP3 gene in South Indian women with idiopathic RPL and to analyze their potential impact on protein structure.

This case-control study involved DNA extraction from 300 participants,including 150 recurrent pregnancy loss (RPL)cases and 150 non-recurrent pregnancy loss (NRPL) controls. Polymerase chain reaction (PCR) and Sanger sequencing were used to identify genetic variants. The identified single-nucleotide polymorphisms (SNPs) were analyzed for frequency differences between the RPL and control groups. Additionally, bioinformatics tools were employed to assess the structural impact of the identified mutations on the FOXP3 protein.

Seven novel single-nucleotide polymorphisms (SNPs) were identified, with four SNPs (-11InsT, 206G>A in exon 2, and 433InsT, 726A>T in exon 7), showing significant frequency variations between RPL and NRPL groups. The modeled structures of FOXP3 apo and mutant proteins displayed similar structural features, including a DNA-binding domain. Molecular dynamics simulation studies revealed comparable stability between the apo and mutant forms of FOXP3.

The identified mutations in the FOXP3 gene can potentially disrupt its critical immune-regulatory functions, leading to impaired immune tolerance during pregnancy, a key factor in the development of RPL. These mutations may alter the activity or stability of regulatory T cells, which are essential for maintaining pregnancy by preventing immune rejection of the fetus.

These findings provide new insights into the genetic underpinnings of idiopathic RPL and underscore the importance of genetic testing for a better understanding of this condition.