Current Protein and Peptide Science - Online First

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.

Autoimmune Thyroiditis (AIT) is caused by defects in the immune system in people with a genetic predisposition to the disease. The most prevalent type of autoimmune thyroiditis is Hashimoto's thyroiditis (HT). The present article reviews the possible relationship between α2-macroglobulin levels and autoantibodies in patients suffering from Hashimoto's disease.

A total of 170 patients with Hashimoto's disease, categorized into subclinical (96 patients) and manifest (74 patients) forms, were enrolled in the study. The control group comprised 65 individuals without thyroid pathologies or other autoimmune diseases. The levels of α2-macroglobulin and autoantibodies, including both organ-specific and non-organ-specific, were determined in all study participants.

Organ-specific antibody and α2-macroglobulin levels were elevated in all patients studied compared to controls. Analysis of organ non-specific antibody levels in patients revealed elevated levels of antibodies to double-stranded (native) DNA in both the subclinical and manifest groups of patients. There were no statistically significant differences in antibody levels to single-stranded (denatured) DNA between the total patient group and the control groups.

The data obtained demonstrated that there is no significant correlation between α2-macroglobulin levels and autoantibody titres, as well as the severity of autoimmune thyroiditis. This finding suggests that α2-macroglobulin may have an unlikely role in the pathogenesis or as a biomarker of disease activity, including in the presence of antibody-dependent cellular damage. Conversely, antibodies directed against double-stranded DNA have exhibited enhanced informativeness and can be regarded as potential markers of the severity of autoimmune thyroid lesions.

Consequently, α2-macroglobulin has no diagnostic value as an indicator of autoimmune process exacerbation in Hashimoto's thyroiditis. Conversely, the presence and level of antibodies to double-stranded DNA may offer a means to assess the severity of the disease and should be the focus of further studies as prognostic markers.

Sialic acid-binding immunoglobulin-like lectins (Siglecs) are cell-surface immunological receptors predominantly expressed on immune cells such as monocytes, macrophages, and dendritic cells. They play a crucial role in regulating inflammatory processes in various diseases and serve as immunological checkpoints in cancer. Despite several immune checkpoint inhibitors targeting Siglecs having entered clinical trials, the number of Siglec-targeted immunotherapies remains limited.

This review aims to investigate the contributions of Siglecs in human diseases and explore novel therapeutic strategies targeting the Siglec-sialic acid immunological axis.

The authors systematically searched PubMed, Web of Science, and Google Scholar for publications mainly from 2015-2025, using search terms related to Siglecs, tumors, autoimmune diseases, and specific Siglec subtypes (CD169, Siglec2). Studies were included if they examined Siglecs biology, immunomodulation, or immunotherapeutic potential. Studies not directly relevant to Siglecs function/therapeutics and non-peer-reviewed materials (conference abstracts, editorials) were excluded. Screening was done via titles and abstracts with data referenced from research article results, and eligible articles underwent full-text review for final inclusion.

The analysis reveals that Siglecs exhibit dual functions, acting as both activators and inhibitors of immune responses. They are implicated in the pathogenesis of various diseases, including cancer, autoimmune disorders, and viral infections. Several Siglec-targeted immunotherapies are currently in clinical trials, demonstrating their potential in disease management. For instance, Siglec15 and Siglec10 have been identified as potential immune checkpoints in cancer, while Siglec2 and Siglec10 play roles in autoimmune diseases like systemic lupus erythematosus (SLE).

Siglecs are key immunomodulators that mediate cell-cell and pathogen interactions, playing pivotal roles in human diseases. Further research into their mechanisms and clinical applications is essential to fully harness their therapeutic potential. Targeting Siglecs offers promising avenues for developing novel immunotherapies, particularly in cancer and autoimmune diseases.

Transforming growth factor-beta (TGF-β) plays a pivotal role in advanced hepatocellular carcinoma (HCC) by modulating immune responses, inflammatory processes, and epithelial-mesenchymal transition (EMT) in hepatocytes. It has emerged as a key therapeutic target for HCC.

This study employs bibliometric analysis to examine literature published between 2000 and 2024, aiming to explore the critical roles of TGF-β in HCC and provide a theoretical foundation for future research.

This study utilized the Web of Science Core Collection (WoSCC) database to analyze publications from January 1, 2000, to October 16, 2024. Visualization tools such as CiteSpace, VOSviewer, and SCImago Graphica were utilized to assess publication trends, countries, institutions, journals, authors, keywords, and references, identifying hotspots, trends, and the evolution of TGF-β research in the context of HCC.

The analysis encompassed 3,026 publications originating from 79 different countries. China was identified as the leading country in publication volume, with Fudan University being the most prolific institution. The journal Hepatology stood out as the leading publication in terms of both the volume of articles and citation influence. Keyword analysis revealed that recent research (2020–2024) has focused on metabolic regulation, the tumor immune microenvironment, and targeted therapies related to the TGF-β signaling pathway in HCC.

This study highlights the publication landscape, research trends, and hotspots of TGF-β-related HCC research from 2000 to 2024, providing valuable insights and a theoretical basis for future studies in this critical field.