Instability Index Research Articles

Defining physicochemical properties of urinary proteins that determine their inhibitory activities against calcium oxalate kidney stone formation

We have recently reported a set of urinary proteins that inhibited calcium oxalate (CaOx) stone development. However, physicochemical properties that determine their inhibitory activities remained unknown. Herein, human urinary proteins were chromatographically fractionated into 15 fractions and subjected to various CaOx crystal assays and identification by nanoLC-ESI-Qq-TOF MS/MS. Their physicochemical properties and crystal inhibitory activities were subjected to Pearson correlation analysis. The data showed that almost all urinary protein fractions had crystal inhibitory activities. Up to 128 proteins were identified from each fraction. Crystallization inhibitory activity correlated with percentages of Ca2+-binding proteins, stable proteins, polar amino acids, alpha helix, beta turn, and random coil, but inversely correlated with number of Ox2−-binding motifs/protein and percentage of unstable proteins. Crystal aggregation inhibitory activity correlated with percentage of stable proteins but inversely correlated with percentage of unstable proteins. Crystal adhesion inhibitory activity correlated with percentage of stable proteins and GRAVY, but inversely correlated with pI, instability index and percentages of unstable proteins and positively charged amino acids. However, there was no correlation between crystal growth inhibitory activity and any physicochemical properties. In summary, some physicochemical properties of urinary proteins can determine and may be able to predict their CaOx stone inhibitory activities.

Isokinetic Strength and Balance Analyses for Predicting Return to Sports After the Latarjet Procedure: A Prospective Cross-sectional Study

Background: Isokinetic torque in shoulder internal rotation (IR) and external rotation (ER) can be considered as potential indicators for dynamic stability of the glenohumeral joint. Purpose: To assess the efficacy of 4-month isokinetic testing in predicting the 6-month return-to-sports (RTS) status after Latarjet surgery, explore its correlations with testing parameters, and identify optimal thresholds to ensure a safe RTS. Study Design: Cohort study; Level of evidence, 2. Methods: The study assessed athletes who underwent the Latarjet stabilization procedure between January 2022 and June 2023. The primary outcome was RTS at 6 months after surgery. The primary examined predictors were isokinetic testing metrics at 4 months postoperatively. Secondary outcomes comprised the modified Closed Kinetic Chain Upper Extremity Stability Test (mCKCUEST) and several patient-reported outcome measures, including the Walch-Duplay score, the Western Ontario Shoulder Instability Index (WOSI), and the Shoulder Instability–Return to Sports after Injury scale. To assess the predictors, patients were divided into those who returned to any level of sports compared with those who did not return to sports. The correlation between isokinetic testing results and other outcome scores was also analyzed. Results: A total of 71 patients (mean age, 27.43 ± 9.09 years) were included in the study. Of these, 23.61% did not return to sports, 38.89% returned at a lower level, and 37.50% returned to the same level. Significant rotational strength disparities were noted. Patients who did not return to sports at 6 months demonstrated inferior strength in concentric ER at 60 deg/s, concentric ER at 240 deg/s, concentric IR at 240 deg/s, and eccentric IR at 30 deg/s (P < .05). Similar trends appeared for all studied patient-reported outcome measures and the mCKCUEST (P < .05). Receiver operating characteristic analysis emphasized the significance of isokinetic testing in concentric ER at 240 deg/s (area under the curve = 0.759; P = .001; cutoff = 0.32 N·m/kg; sensitivity = 100.0%; specificity = 49.1%) and eccentric ER at 30 deg/s (area under the curve = 0.760; P = .001; cutoff = 0.51 N·m/kg; sensitivity = 94.1%; specificity = 49.1%) for RTS prediction. Additionally, ER strength moderately correlated with the Walch-Duplay score across all examined velocities (r = 0.26-0.34; P < .05). The modified WOSI score was weakly linked to ER strength at 240 deg/s and 30 deg/s (r = 0.24-0.25; P < .05) as well as moderately linked to the limb symmetry index in ER at 60 deg/s and 30 deg/s (r = 0.30-0.38; P < .05). Conclusion: Isokinetic testing can act as an independent predictor of successful RTS after Latarjet surgery, with concentric ER at 240 deg/s, concentric IR at 240 deg/s, eccentric ER at 30 deg/s, and eccentric IR at 30 deg/s showing the most accuracy. Strength recovery in ER was associated with better Walch-Duplay and modified WOSI scores.

B-cell epitope prediction of MPB83 protein as a candidate for serodiagnostic antigen of bovine tuberculosis in human: In silico study

Background: Bovine tuberculosis (bTB) can be transmitted to humans by inhalation or consumption of incomplete pasteurized milk and dairy products derived from infected cows. Most cases of Mycobacterium tuberculosis (M. bovis) infection are resistant to tuberculosis (TB) drugs. The risk of death during treatment for bTB has been reported to be 2.55 times higher than for TB. However, the quality of diagnostic methods for bTB remains relatively low.Objective: We aim to evaluate the potential of the B-cell epitope of the MPB83 protein as a candidate bTB serodiagnostic antigen using an in silico approach. Methods: This study was a computer-based descriptive study using secondary data from the National Center for Biotechnology Information (NCBI) protein database. The MPB83 protein sequence was obtained from M. tuberculosis variant bovis AF2122/97 from the United Kingdom. We described the characteristics of the linear epitope of the M. bovis B-cell protein MPB83 by measuring antigenicity, molecular weight, instability index, and Grand Average of Hydropathy (GRAVY) score. The tools used in this study were IBIVU PRALINE, VaxiJen v2.0, IEDB, ExPASy ProtParam, Cluspro, and the PyMOL application.Results: We found an epitope that could be used for bTB serodiagnostic antigen with low conservation, the 106KLNPDVNLVDTLN118 epitope. It has the molecular weight, instability index, and GRAVY score of 1638.76 Da, -28.44, and -0.300, respectively. Epitopes with the best criteria were simulated by docking to human major histocompatibility complex (MHC) class II. Docking results showed that the lowest binding energy was -644.8 kcal/mol. Further analysis using the PyMOL application obtained 14 hydrogen bonds with bond distances ranging from 1.7 Å to 2.2 Å, all of which showed strong hydrogen bonds.Conclusion: The B-cell epitope of MPB83 protein sequence 106KLNPDVNLVDTLN118 has a potential serodiagnostic antigen candidate for human bTB.

Rational Design of a Multi-epitope Vaccine Using Neoantigen Against Colorectal Cancer Through Structural Immunoinformatics and ML-Enabled Simulation Approach.

Colorectal cancer poses a substantial global health burden. Regarding WHO, the global burden of colorectal cancer will be about 3.2 million new cases by the year 2040. Simultaneously, it indicated that this cancer will cause 6 million deaths per year. Despite advancements in chemotherapy and monoclonal antibody therapy, the disease remains a significant challenge due to the resistance of cancer stem cells. This study endeavors to design a multi-epitopic peptide (9-mer epitopes) neoantigen-based vaccine targeting the TLR4/MD2 complex as a potential vaccine candidate. These tumor-specific neoantigens (TSA) are considered novel antigens that can be used for vaccine development against cancer. To develop the neoantigen vaccine candidate, we used the SPENCER database, and 140 lncRNA-derived epitopes were retrieved. From 140 epitopes, we selected seven neoantigens with high antigenic properties for the vaccine construct. A novel vaccine containing epitopes, linkers(EAAAK and CPCPG), and adjuvants(ribosomal [50S]protein L7L12) was formulated utilizing immunoinformatics tools. The vaccine's biophysical properties were evaluated, revealing its antigenicity(0.6469), stability(instability index:37.05), and potential for immune system interaction. In-depth structural analyses, molecular docking studies, and ML-enabled immune simulation profiling underscored the vaccine's structural integrity, binding affinity with TLR4, and ability to elicit robust immune responses against colorectal cancer antigens. These findings suggest that the multi-epitopic vaccine holds promise as a next-generation approach to combat colorectal cancer. Our in silico studies exhibit potentiality of the vaccine candidate; however, further in vivo and in vitro investigations are crucial to validate immunogenicity, safety, and efficacy before clinical implementation. Our study developed a first-time lncRNA-derived neoantigen-based cancer vaccine.

Comparison of Modified Latarjet Procedure With Bone Graft Versus Standard Latarjet Procedure: Evaluating Outcomes and Stability.

The Latarjet procedure is a well-established technique for managing repeated anterior shoulder dislocation accompanied by massive glenoid bone loss. Aim of this article was to assess outcomes amongmodified Latarjet procedure using allograft from Iliac bone and a standard Latarjet procedure using the coracoid process. Six patients with recurrent anterior shoulder instability and significant glenoid bone loss were retrospectively analyzed. Three patients underwent the modified Latarjet procedure with iliac crest bone graft (Group A), and three underwent the standard Latarjet procedure (Group B). Outcomes were assessed at the 12-month follow-up, including shoulder stability, functional scores (Constant, the American Shoulder and Elbow Surgeons (ASES) score, and the Western Ontario Shoulder Instability Index (WOSI)), range of motion, complications, and return to sports. Both groups showed improvements in stability and functional scores, with no recurrent instability reported. Group A demonstrated slightly higher functional scores and range of motion. One patient in Group A experienced donor site pain, while one patient in Group B showed significant graft resorption. Graft union was achieved in all patients. Two-thirds of patients in each group returned to their pre-injury level of sports participation. Both techniques provided good clinical outcomes for anterior shoulder instability with glenoid bone loss. The modified Latarjet with iliac crest graft may offer slight advantages in functional outcomes and graft preservation but is associated with potential donor site morbidity. Larger, prospective studies are needed to definitively compare these techniques.

Glenolabral Articular Disruption (GLAD) Is Not Associated with Worse Outcomes or Higher Instability Recurrence after Arthroscopic Bankart Repair-A Matched-Pair Analysis.

Background: This study aimed to compare clinical outcomes and recurrence of instability after arthroscopic Bankart repair (ABR) in patients with anterior shoulder instability, with and without a GLAD lesion, while distinguishing between primary and recurrent instability. Methods: Consecutive patients who underwent isolated ABR between January 2012 and December 2021 were included. Patients with a concomitant GLAD lesion were matched in with patients without a GLAD lesion according to the following criteria: age, sex, BMI, follow-up time, and primary versus recurrent instability. At minimum two-year follow-up, the clinical outcome (Rowe score, redislocation rate) and the functional outcome, including the American Shoulder and Elbow Surgeons (ASES) score, Western Ontario Shoulder Instability Index (WOSI), Oxford Shoulder Instability Score (OSIS), satisfaction (1-10 scale, 0 = unsatisfied, 10 = very satisfied), and Visual Analogue Scale (VAS), were compared between groups. Results: In total, 28 patients (14 GLAD vs. 14 Bankart; age: 32.5 ± 13.0 years; sex: 92.9% male; BMI: 24.6 ± 2.2) were included 6.9 ± 2.8 (2-11) years after isolated ABR (follow-up rate 63.6%). Clinical and functional outcome did not differ significantly between patients with versus without GLAD lesions (ASES score: 100 [96.5-100] vs. 97.5 [93.3-100], p = 0.27); WOSI (%): 9.0 [3.7-24.5] vs. 3.8 [0.8-8.9], p = 0.22; Rowe score: 90.0 [75.0-100] vs. 95.0 [78.8-100], p = 0.57; OSIS: 46 [44.7-48] vs. 46 [43.0-48], p = 0.54; satisfaction: 8.9 ± 1.4 vs. 8.0 ± 1.4, p = 0.78; VAS 0 [0-1.3] vs. 0 [0-1.0]. In both groups, two patients (14.3%) reported a redislocation during the observation period. Conclusions: At short- to mid-term follow-up, ABR showed favorable outcomes, low dislocation rates, and high patient satisfaction, regardless of the presence of a GLAD lesion or primary versus recurrent instability. However, follow-up time was heterogeneous, and the follow-up rate was marginal.

Chromosomal copy number based stratification of gastric cancer has added prognostic value to Lauren’s histological classification

BackgroundThe Cancer Genome Atlas (TCGA) recognizes four molecular subgroups of gastric cancer: Epstein-Barr virus (EBV) positive, microsatellite instable (MSI), genomically stable (GS), and chromosomal instable (CIN). Since a GS/CIN classifier is lacking, alternative markers such as Lauren’s histopathology or CDH1/p53 immunohistochemistry are commonly applied. Here we compared survival of gastric cancer subgroups determined by four methods.Methods309 EBV negative and microsatellite stable tumors were included from the Dutch D1/D2 trial and assigned to subgroups by: (i) TCGA’s specific chromosomal copy number aberrations, (ii) genome instability index (GII), (iii) Lauren’s classification, and (iv) CDH1/p53 immunohistochemistry. Subgroups were associated with cancer-related survival (CRS).ResultsFive-year CRS was 42.0% for diffuse and 49.5% for patients with intestinal type tumors, and 57.8% for GS and 41.6% for patients with CIN tumors. Classification by GII or CDH1/p53 IHC did not correlate with CRS. The combination of TCGA and Lauren classifications resulted in four distinct subgroups. Five-year CRS for GS-intestinal (n = 24), GS-diffuse (n = 57), CIN-intestinal (n = 142) and CIN-diffuse (n = 86) was 61.4%, 56.5%, 47.6%, and 31.5%, respectively.ConclusionsTCGA’s GS and CIN subgroups have additional prognostic value to Lauren’s classification in resectable gastric cancer. GS-intestinal, GS-diffuse, CIN-intestinal and CIN-diffuse are suggested stratification variables for future studies.

P38. Correlating dynamic instability indices to degenerative findings on MRI in patient with chronic low back pain

P38. Correlating dynamic instability indices to degenerative findings on MRI in patient with chronic low back pain

Auto-expansion of in vivo HDAd-transduced hematopoietic stem cells by constitutive expression of tHMGA2

We developed an invivo hematopoietic stem cell (HSC) gene therapy approach that does not require cell transplantation. To achieve therapeutically relevant numbers of corrected cells, we constructed HSC-tropic HDAd5/35++ vectors expressing a 3' UTR truncated HMGA2 gene and a GFP reporter gene. A SB100x transposase vector mediated random integration of the tHMGA2/GFP transgene cassette. HSCs in mice were mobilized by subcutaneous injections of G-CSF and AMD3100/Plerixafor and intravenously injected with the integrating tHMGA2/GFP vector. This resulted in a slow but progressive, competitive expansion of GFP+ PBMCs, reaching about 50% by week 44 with further expansion in secondary recipients. Expansion occurred at the level of HSCs as well as at the levels of myeloid, lymphoid, and erythroid progenitors within thebone marrow and spleen. Importantly, based on genome-wide integration site analyses, expansion was polyclonal, without any signs of clonal dominance. Whole-exome sequencing did not show significant differences in the genomic instability indices between tHGMGA2/GFP mice and untreated control mice. Auto-expansion by tHMGA2 was validated in humanized mice. This is the first demonstration that simple injections of mobilization drugs and HDAd vectors can trigger auto-expansion of invivo transduced HSCs resulting in transgene-marking rates that, theoretically, are curative for hemoglobinopathies.

Price Dynamics and Integration of Onion Markets in India

The current study uses the Correlation matrix, Vector Auto Regressive model, Granger causality test and Instability index to investigate the level of market integration, price transmission and price instability in the key onion markets in India, namely, Azadpur, Bangalore, Belgaum, Hassan, Haveri, Hubli, Mysore, and Nashik. The findings provided substantial evidence for the co-integration and interdependence of these markets. Bangalore market has a positive relationship with the Nashik and Azadpur markets, with coefficients of 1.15 and 0.54, respectively. Bangalore and Belgaum displayed a solid, significant positive correlation with a coefficient of 0.770. Conversely, some markets show relatively weaker correlations, such as Azadpur and Hassan, with a coefficient of 0.389. Instability across major onion markets revealed that all the markets are volatile, as the instability index was more than 30 per cent. Since there is unidirectional causality from Hubli-Bangalore and Azadpur-Bangalore market pairs, it makes it necessary for future research to examine the influence of internal and external factors like market infrastructure, improving information technology to allow regular flow of market information to assist farmers in increasing their income. Keywords: Granger causality, instability, price transmission, price volatility. JEL Codes: B23, C01, C22, E31.

Evaluation of heavy hydrocarbon liquid in the Eastern Dahomey Basin, Southwestern Nigeria (case study of Agbabu Bitumen in Ondo State)

Solvent-aided dilution processes are often employed to recover bitumen by reducing its viscosity. In this study, methanol, toluene, and xylene were investigated as potential hydrocarbon solvents for solvent-aided hydrocarbon recovery of Agbabu bitumen. To achieve this, the solubility of the sample in the solvents was determined with a solubility test, its viscosity was measured with the Saybolt Furol Viscometer using the ASTM D88 method, and its SARA analysis was conducted using high-performance liquid chromatography. Agbabu bitumen was found to have a high content of saturates and aromatics. Viscosity decreases as pressure increases, while solubility reduces as temperature increases. The sample was most soluble in toluene, and its viscosity was reduced most in it, followed by xylene. Methanol reduced the saturates content and significantly raised the asphaltene content, keeping the mixture viscosity high. In terms of asphaltene stability of the mixtures, the bitumen-methanol system with the lowest colloidal instability index of 0.874 will give a mild asphaltene deposit issues compared with others that are unstable. This approach of combining multiple tests with the CII can accurately predict the behavior of Agbabu bitumen in solvents and enhance decision on the choice of bitumen recovery technology.

Influence of Nitrogen Fertilization on Quality Parameters and Heat Use Efficiency of Basmati Rice

ABSTRACT Physical, milling, cooking, and chemical qualities of basmati cultivars had great concern for researchers, producers, traders, and consumers as instability index of production is higher. Quality may be enhanced by good N optimization to responsive basmati varieties. Therefore, a field experiment was conducted for two consecutive years under split-plot design. Results revealed that application of 120 kg N ha−1 improved heat use efficiency (4.85 & 4.91 kg ha−1 degree-day−1), milling quality traits, physical quality, and chemical quality while aroma score (up to 2.50) improved only up to 60 kg N ha−1. Highest protein (7.79 & 7.77%) and least amylose (21.88 & 22.24%) content were recorded with highest N rates while highest protein yield (414.18 & 402.66 Kg ha−1) reported at 120 Kg N ha−1. Basmati varieties, HUBR 10–9 and HUBR 2–1, were superior in most of quality traits along heat use efficiency (4.62–4.70 kg ha−1 degree-day−1). However, short-grained aromatic variety HUR 917 recorded higher head rice recovery (64.34 & 63.21%), protein (7.61 & 7.63%), and amylose content over others. Most important trait, i.e. aroma (1.83–2.38) score, was higher with basmati varieties. Overall, basmati (HUBR 10–9 and HUBR 2–1) grown with 120 kg N ha−1 proved superiority. This study achieved better heat use efficiency along with improved quality of basmati rice which found to be positively correlated with most quality traits of basmati rice. The low amylose in basmati is preferable, and amylose found to be negatively correlated with heat use efficiency.

Influence of Tis108 on GA content and expression of key enzyme GeCYP714A1 involved in GA deactivation of Gastrodia elata

To investigate the influence of the strigolactone inhibitor Tis108 on the growth of Gastrodia elata, this study treated G. elata tuber with Tis108 solution of 10 μmol·L~(-1) and measured the content of endogenous hormone gibberellin(GA) in the tuber. By using reverse transcription-polymerase chain reaction(RT-PCR) technology, the key enzyme GeCYP714A1 gene involved in GA deactivation was cloned. Bioinformatics analysis on the GeCYP714A1 gene was carried out by using ExPASy, SWISS-MODEL, MEGA, etc., and its expression levels in different parts of G. elata were determined. The results showed that after Tis108 treatment, GA content in G. elata tuber was significantly increased, and the transcription level of the GeCYP714A1 gene was significantly decreased. The full length of the coding region of the GeCYP714A1 gene is 1 173 bp, encoding 390 amino acids. The protein has a molecular weight of 44.85 kDa, a theoretical isoelectric point of 9.83, an instability index of 49.20, an aliphatic index of 89.03, and a grand average of hydropathicity of-0.235, classifying it as an unstable, basic, hydrophilic protein, and the GeCYP714A1 protein was localized in the mitochondria, lacking a signal peptide and a transmembrane structure. Phylogenetic tree analysis revealed that GeCYP714A1 was most closely related to the DcCYP714C2(PKU78454.1) protein from Dendrobium candidum, with a sequence identity of 67.25%. The qRT-PCR analysis of the expression patterns of the GeCYP714A1 gene indicated that GeCYP714A1 had the highest transcription level in G. elata tuber, followed by stem and inflorescence. The study represented that Tis108 inhibited the transcription level of GeCYP714A1 involved in GA deactivation in G. elata tuber, thereby increasing the accumulation of GA and affecting the growth of G. elata tuber. These results provided a basis for further studies of strigolactone regulation of GA signal and tuber development in G. elata.

Outcomes following arthroscopic treatment of off-track Hill-Sachs lesions using fresh osteochondral allograft plugs: a case series

BackgroundOsteochondral allograft (OCA) transplantation is a well-described technique for the treatment of large, engaging Hill-Sachs lesions (HSLs). Traditionally, OCAs are size-matched to the defect, which can be expensive and time-consuming, and the majority of described techniques require an open approach. Recently, an all-arthroscopic approach to Hill-Sachs OCA transplantation using premade plugs was described, eliminating the need for size-matching and graft harvest. Hypothesis/PurposeThe purpose of this study was to evaluate postoperative outcomes of patients who have undergone arthroscopic treatment of HSLs using premade OCA plugs. We hypothesized that these patients would have improved pain and function without recurrent instability. MethodsA retrospective chart review was performed using operative reports for a single surgeon with search terms “Hill-Sachs” and “allograft.” Patients were excluded if an open approach was used or if graft harvest was performed. Postoperative imaging was reviewed to assess for graft incorporation and reconstitution of the HSL. Recurrent instability and reoperation were recorded. Patients completed surveys including the American Shoulder and Elbow Surgeons (ASES) score and the Western Ontario Shoulder Instability Index (WOSI). ResultsFive patients were identified through chart review and met inclusion criteria. All patients underwent concurrent labral repair and two patients underwent a concurrent open Latarjet procedure. Postoperative radiographs showed reconstitution of the HSLs in all patients. There were no complications in the postoperative period with no recurrent instability or reoperations in any patient. The average ASES score was 87% (higher score indicating better outcome) with standard deviation 9.7, and the average WOSI score was 27% (lower score indicating better outcome) with standard deviation 8.3. ConclusionFavorable outcomes can be expected after arthroscopic treatment of Hill-Sachs lesions using premade OCA plugs. Further research is needed to assess larger patient cohorts and compare outcomes to size-matched approaches.

Enhancing the stability of O/W emulsions by the interactions of casein/carboxymethyl chitosan and its application in whole nutrient emulsions

The influence of Carboxymethyl chitosan (CMCS) on the emulsification stability mechanism of casein (CN) and its effects on the stability of whole nutrient emulsions were investigated. The complex solutions of CN and CMCS were prepared and the turbidity, ultraviolet (UV) absorption spectrum, fluorescence spectrum, circular dichroism (CD) spectrum, Fourier transform infrared (FTIR) spectrum, interfacial tension and microstructural observations were used to study the inter-molecular interaction of CMCS and CN. The effects of CMCS on the emulsion stability of CN were further analyzed by particle size, ζ-potential, instability index and rheological properties. Moreover, the accelerated stability of whole nutrient emulsions prepared by CMCS and CN was evaluated. The results revealed that CN-CMCS complexes were mainly formed by hydrogen bonding. The stability of the CN-CMCS composite emulsions were improved, as evidenced by the interfacial tension decreasing from 165.96 mN/m to 158.49 mN/m, the particle size decreasing from 45.85 μm to 12.98 μm, and the absolute value of the potential increasing from 29.8 mV to 33.5 mV. The stability of whole nutrient emulsion was also significantly enhanced by the addition of CN-CMCS complexes. Therefore, CN-CMCS complex could be served as a novel emulsifier to improve the stability of O/W emulsions.

Growth and instability in mungbean production in India

Mungbean is mainly cultivated in East Asia, Southeast Asia, and the Indian subcontinent. It is the third important pulse crop of India grown in nearly 18 percent of the total pulse area. The paper delves into the current state of mungbean in India, examining the shift of the area of mungbean across zones and its underlying reasons. The study has been carried out based on secondary data and the data was collected for the period from 1970-71 to 2020- 21. Compound annual growth rate and instability index were computed. The results showed that mungbean area and production declined in the eastern zone but showed considerable growth in the southern zone. Production in all the major states increased with a positive growth rate except Odisha. In Odisha, Rajasthan and Madhya Pradesh areas have shown higher instability than productivity, but Karnataka, Maharashtra, and all of India’s productivity was found more unstable. Therefore, it is necessary to promote the adoption of conservation practices, harvesting of rainwater by creation of structures like farm ponds as well as to expand irrigation facilities to overcome variability in productivity.

Molecular analysis of recombinant collagenase from Bacillus siamensis strain Z1: Gene Cloning, expression and in-silico characterization

This study focuses on gene cloning, expression, biochemical and analytical characterization along with structural and functional characterization of collagenase followed with molecular docking, dynamics study and Molecular Mechanics Poisson-Boltzmann Surface Area (MMPBSA). The collagenase gene identified from the genome of the novel collagenase Bacillus siamensis strain Z1 is introduced into E.coli DH5α, subsequently expressed in E.coli BL21 (DE3) withisopropyl β −d – 1 − thiogalactopyranoside(IPTG) induction and further affinity purified yielding in ∼ 89.4 kDa recombinant collagenase which demonstrated alkali characteristics and thermostability determined by thermodynamic parameters. Recombinant collagenase revealed good stability when exposed to diverse biochemical components. The recombinant collagenase identity was confirmed through matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) showing specific mass peaks and via N-terminal sequence analysis as MTAVNQTISK. Moreover, the concluded N-terminal amino acid sequence from Edman degradation displayed significant resemblance. The structural and functional analysisof recombinant collagenase was analysed by Circular dichroism (CD), Proton nuclear magnetic resonance (1H NMR) spectrometry and Thermogravimetric analysis (TGA). The recombinant collagenase also showed gelatin liquefaction ability. The collagenase gene sequence is also assessed for structural and functional characterizations by using various computational tools and revealed its classification in U32 family peptidase. A Grand average of hydropathicity index (GRAVY) score of −0.295 and instability index of 37.22 was obtained. Homology model of collagenase gene was generated by employing SWISS-MODEL and structure analysis by Ramachandran plot. Molecular docking of modelled collagenase with four different substrates was carried out by PyRx and Autodocking. Highest docking score of −12.7 kcal/mol was obtained for Alaska pollock hydroxyproline containing marine collagen peptide (APHCP). Subsequently, Molecular dynamics and simulations for highest score docked complex was assessed using GROningenMAchine for Chemical Simulations (GROMACS).

Effect of non-covalent binding of tannins to sodium caseinate on the stability of high-internal-phase fish oil emulsions

In this study, we designed the noncovalent binding of sodium caseinate (SC) to tannic acid (TA) to stabilize high internal phase emulsions (HIPEs) used as fish oil delivery systems. Hydrogen bonding was the dominant binding force, followed by weak hydrophobic interaction and weak van der Waals forces, as demonstrated by FTIR, fluorescence spectroscopy, and molecular docking experiments, with a binding constant of 3.25 × 106, a binding site of 1.2, and a static quenching of the binding. Increasing SC:TA from SC to 2:1 decreased the particle size from 107.37 ± 10.66 to 76.07 ± 2.77 nm and the zeta potential from −6.99 ± 2.71 to −22 ± 2.42 mV. TA increased the interfacial tension of SC, decreased the surface hydrophobicity from 1.3 × 104 to 1.6 × 103 and improved the oxidation resistance of SC. The particle size of high internal phase emulsions stabilized by complexes with different mass ratios (SC:TA from 1:0 to 2:1) increased from 4.9 ± 0.02 to 12.9 μm, the potential increased from −32.37 ± 2.7 to −35.07 ± 2.58 mV, and the instability index decreased from 0.75 to 0.02. Thicker interfacial layers could be observed by laser confocal microscopy, and an increase in the storage modulus indicated a formation of a stronger gel network. SC:TA of 1:0 showed emulsion breakage after 14 d of storage at room temperature. SC:TA of 2:1 showed the lowest degree of oil-water separation after freeze-thaw treatment. Especially, the most stable high endo-phase emulsion (at SC:TA of 2:1) prepared at each mass ratio was selected for further stability exploration. The emulsion particle size increased only from 15.63 ± 0.06 to 22.27 ± 0.35 μm at salt ion concentrations of 50–200 mM and to 249.33 ± 31.79 μm at 300 mM. The instability index and storage modulus of the high endo-phase emulsions increased gradually with increasing salt ion concentrations. At different heating temperatures (55–85 °C), the instability index of the high internal phase emulsion gradually decreased and the storage modulus gradually increased. Meanwhile, at 50 °C for 15 d of accelerated oxidation, the content of hydroperoxide decreased from 53.32 ± 0.18 to 37.48 ± 0.77 nmol/g, about 29.7 %, and the thiobarbituric acid value decreased from 1.06 × 103 to 0.8 × 103, about 24.5 %, in the high endo-phase emulsions prepared by 2:1 SC:TA compared to the fish oils, and the SC-stabilized high endo-phase only emulsion broke at the sixth day of oxidation. From the above findings, it was concluded that the high internal phase emulsion prepared with SC:TA of 2:1 can be used as a good delivery system for fish oil.

Balance Control Impairments in Usher Syndrome.

To explore postural disability in Usher Syndrome (USH) patients using temporal posturographic analysis to better elucidate sensory compensation strategies of deafblind patients for posture control and correlate the Activities-specific Balance Confidence (ABC) scale with posturographic variables. Thirty-four genetically confirmed USH patients (11 USH1, 21 USH2, 2 USH 4) from the Otolaryngology Outpatient Clinic and 35 controls were prospectively studied using both classical and wavelet temporal analysis of center of pressure (CoP) under different visual conditions on static and dynamic platforms. The functional impact of balance was assessed with the ABC scale. Classical data in the spatial domain, Sensorial Organization Test, and frequency analysis of the CoP were analyzed. On unstable surfaces, USH1 had greater CoP surface area with eyes open (38.51 ± 68.67) and closed (28.14 ± 31.64) versus controls (3.31 ± 4.60), p < 0.001 and (7.37 ± 7.91), p < 0.001, respectively. On an unstable platform, USH consistently showed increased postural sway, with elevated angular velocity versus controls with eyes open (USH1 [44.94 ± 62.54]; USH2 [55.64 ± 38.61]; controls [13.4 ± 8.57]) (p = 0.003; p < 0.001) and closed (USH1 [60.36 ± 49.85], USH2 [57.62 ± 42.36]; controls [27.31 ± 19.79]) (p = 0.002; p = 0.042). USH visual impairment appears to be the primary factor influencing postural deficits, with a statistically significant difference observed in the visual Sensorial Organization Test ratio for USH1 (80.73 ± 40.07, p = 0.04) and a highly significant difference for USH2 (75.48 ± 31.67, p < 0.001) versus controls (100). In contrast, vestibular (p = 0.08) and somatosensory (p = 0.537) factors did not reach statistical significance. USH exhibited lower visual dependence than controls (30.31 ± 30.08) (USH1 [6 ± 11.46], p = 0.004; USH2 [8 ± 14.15], p = 0.005). The postural instability index, that corresponds to the ratio of spectral power index and canceling time, differentiated USH from controls on unstable surface with eyes open USH1 (3.33 ± 1.85) p < 0.001; USH2 (3.87 ± 1.05) p < 0.002; controls (1.91 ± 0.85) and closed USH1 (3.91 ± 1.65) p = 0.005; USH2 (3.92 ± 1.05) p = 0.045; controls (2.74 ± 1.27), but not USH1 from USH2. The canceling time in the anteroposterior direction in lower zone distinguished USH subtypes on stable surface with optokinetic USH1 (0.88 ± 1.03), USH2 (0.29 ± 0.23), p = 0.026 and on unstable surface with eyes open USH1 (0.56 ± 1.26), USH2 (0.072 ± 0.09), p = 0.036. ABC scale could distinguish between USH patients and controls, but not between USH subtypes and it correlated with CoP surface area on unstable surface with eyes open only in USH1(ρ = 0.714, p = 0.047). USH patients, particularly USH1, exhibited poorer balance control than controls on unstable platform with eyes open and appeared to rely more on proprioceptive information while suppressing visual input. USH2 seems to use different multisensory balance strategies that do not align as well with the ABC scale. The advanced analysis provided insights into sensory compensation strategies in USH subtypes.

Multiomics unveils extracellular vesicle-driven mechanisms of endothelial communication in human carotid atherosclerosis.

Background: Carotid atherosclerosis is orchestrated by cell-cell communication that drives progression along a clinical continuum (asymptomatic to symptomatic). Extracellular vesicles (EVs) are cell-derived nanoparticles representing a new paradigm in cellular communication. Little is known about their biological cargo, cellular origin/destination, and functional roles in human atherosclerotic plaque. Methods: EVs were enriched via size exclusion chromatography from human carotid endarterectomy samples dissected into paired plaque and marginal zones (symptomatic n=16, asymptomatic n=13). EV cargos were assessed via whole transcriptome miRNA sequencing and mass spectrometry-based proteomics. EV multi-omics were integrated with bulk and single cell RNA-sequencing (scRNA-seq) datasets to predict EV cellular origin and ligand-receptor interactions, and multi-modal biological network integration of EV-cargo was completed. EV functional impact was assessed with endothelial angiogenesis assays. Results: Carotid plaques contained more EVs than adjacent marginal zones, with differential enrichment for EV-miRNAs and EV-proteins in key atherogenic pathways. EV cellular origin analysis suggested that tissue EV signatures originated from endothelial cells (EC), smooth muscle cells (SMC), and immune cells. Integrated tissue vesiculomics and scRNA-seq indicated complex EV-vascular cell communication that changed with disease progression and plaque vulnerability (i.e., symptomatic disease). Plaques from symptomatic patients, but not asymptomatic patients, were characterized by increased involvement of endothelial pathways and more complex ligand-receptor interactions, relative to their marginal zones. Plaque-EVs were predicted to mediate communication with ECs. Pathway enrichment analysis delineated an endothelial signature with roles in angiogenesis and neovascularization - well-known indices of plaque instability. This was validated functionally, wherein human carotid symptomatic plaque EVs induced sprouting angiogenesis in comparison to their matched marginal zones. Conclusion: Our findings indicate that EVs may drive dynamic changes in plaques through EV- vascular cell communication and effector functions that typify vulnerability to rupture, precipitating symptomatic disease. The discovery of endothelial-directed angiogenic processes mediated by EVs creates new therapeutic avenues for atherosclerosis.