Biotechnology and applied biochemistry最新文献

This study examines the disparities between people suffering from chronic lower back pain (LBP) and those who do not, with a particular focus on how extended periods of sitting with poor posture affect fatigue and discomfort in the sternocleidomastoid (SCM) muscles among office employees. Eighty university staff and students were enrolled in the study by matching age, BMI, and type of job. They were split into two groups later: a control group (n = 40) with no back pain and a pain group (n = 40) with a history of lower back pain (LBP). Pain intensity was measured using a Visual Analog Scale (VAS), while SCM muscle activity was measured via surface electromyography (sEMG) during both normal and hunched postures. Participants maintained each posture for a specified duration of 30 min. The study revealed that Pain_Hunched group exhibited significantly reduced SCM muscle activity compared to the Control_Hunched group (mean difference = -9.728, p < 0.001). Furthermore, the SCM muscle activity in the Pain_Hunched group was significantly lower than that of the Pain_Normal group (mean difference = -2.769, p = 0.041). These results highlight the heightened SCM muscle activity during hunched postures, particularly in individuals with LBP. The results emphasize the crucial role posture plays in influencing SCM muscle activation and pain perception among individuals with LBP. These results imply that correcting one's posture could be a useful pain management intervention technique for this population. The long-term impacts of postural adjustments and their possible advantages in clinical contexts, such as incorporating ergonomic interventions or specific exercise regimens, should be investigated in future studies.

Pyridoxal 5'-phosphate (PLP) plays an essential role in a multitude of cellular processes due to its function as a critical coenzyme. This study introduces a significant advancement in PLP biosynthesis by enhancing the stability and activity of Escherichia coli-derived pyridoxal kinase (EcPdxK) through immobilization on an innovative epoxy resin, LXTE-600. Our approach involved the systematic optimization of enzyme loading, coupling duration, and temperature, which resulted in improved immobilization efficiency and a high loading capacity of 80 mg/g. The characterization of immobilized EcPdxK@LXTE-600 was conducted using Fourier transform infrared spectroscopy (FTIR) and confocal laser scanning microscopy (CLSM), confirming successful immobilization. This process notably enhanced the enzyme's performance, increasing its tolerance to pH and temperature fluctuations, thereby improving its thermal stability. The immobilized EcPdxK@LXTE-600 retained over 80% of its initial activity after 4 weeks of storage at 4°C and could be reused up to eight cycles while maintaining more than 70% of its initial activity. These findings not only demonstrate the efficacy of the LXTE-600-based immobilization method but also suggest promising industrial applications for the sustainable production of PLP, potentially revolutionizing approaches in biotechnological and pharmaceutical sectors.

The objective of the study was to investigate the value of platelet distribution width (PDW) as a prognostic biomarker by comparing PDW between COPD patients with pulmonary hypertension (PH) (PASP 50 mmHg) and those without PH (PASP < 50 mmHg) and to explore its diagnostic and predictive value in this population. A retrospective study of 270 COPD patients in Liaocheng People's Hospital (July 2018 to July 2019) was done by dividing them into two groups according to pulmonary artery systolic pressure (PASP): COPD-only (PASP <50 mmHg) and COPD with PH (PASP ≥50 mmHg). Routine blood tests, C-reactive protein, pulmonary function tests, echocardiography, Chronic Obstructive Pulmonary Disease Assessment Questionnaire (CAT), Clinical COPD Questionnaire were performed. PDW was higher in COPD with PH group than COPD-only group. After adjusting for confounders including age, smoking history, CAT scores, white blood cell count, PDW, and NT-proBNP in COPD with PH, PDW was positively correlated with various parameters. PDW can diagnose COPD with PH, also prognostic value and cardiovascular distinction in these patients. The study concluded that PDW is a prognostic marker, reflecting pulmonary and cardiovascular physiology in COPD patients with PH. It can be used for early diagnosis, risk stratification, and individualized management for this high-risk population.

Obesity, modifiable and an avertable medical condition, has become a global threat due to rapid modernization and industrialization. Swift growth in modernization not only eases the day-to-day life, it also mandates sedentary lifestyle, which leads to various noncommunicable diseases. At present one in eight people in global population are obese, and these booming obese individuals are prone to various other micro- and macrovascular diseases such hyperglycemia, myocardial infraction, hypertension, stroke, and so forth. Ample research had unveiled an intricate association perceived between obesity and Type 2 diabetes mellitus pathogenesis. Although the intake of anti-obesity drugs along with anti-diabetic drugs had effectively regulated the hyperglycemic conditions in diabetic patients, it causes various side effects on long-term usage. Coumarins are phytochemicals that have demonstrated pharmacological properties including anti-inflammatory, antioxidant, anti-tumor, and so forth. In this analysis, we assessed anti-obesity and anti-diabetic potency of aesculetin, a courmarin derivative. The rats were induced obesity with high-fat diet and subjected to streptozotocin infusion to induce hyperglycemia. Obese diabetic induced rats were treated with aesculetin and assessed for its anti-diabetic effect. BMI were assessed in the rats to analyze the anti-obesity effect of aesculetin. Diabetic profile test and lipid profile test were performed to evaluate the anti-diabetic effect of aesculetin. Ameliorative effect of aesculetin in obese rats during hyperglycemic conditions was assessed with renal profile test, hepatic function biomarkers, and by histopathological analysis of cardiac tissue. The anti-inflammatory and antioxidant property were also assessed to determine the mechanism of action of aesculetin. To confirm the anti-obesity potency of aesculetin, adipokines levels were estimated. Aesculetin eminently decreased the BMI, HbA1c, cholesterol levels, and intensified secretion of insulin in obese diabetic rats. It also regulated the renal, hepatic functional markers and prevented cardiac tissue injury in obese diabetic rats. It regulated the adipokines, increased antioxidants, and decreased level of proinflammatory cytokines, thereby prevented obesity-induced hyperglycemic effects in rats. To conclude, our findings had confirmed the supplementary intake of aesculetin prevents obesity-induced hyperglycemic disorder in rats.

Lung squamous cell carcinoma (LUSC) is a malignancy with limited therapeutic options. Immunogenic cell death (ICD) has the potential to enhance the efficacy of cancer therapy by triggering immune responses. We aimed to explore the potential of ICD-based classification in predicting prognosis and response to immunotherapy for LUSC. RNA-seq information and clinical data of LUSC patients were obtained from The Cancer Genome Atlas (TCGA) dataset. ICD-related gene expressions in LUSC samples were analyzed by consensus clustering. Subsequently, differentially expressed genes (DEGs) between different ICD-related subsets were analyzed. Tumor mutation burden, immune cell infiltration, and survival analyses were conducted between different ICD subsets. Finally, an ICD-related risk signature was constructed and evaluated in LUSC patients, and the immunotherapy responses based on the gene expressions were also forecasted. ICD-high and ICD-low groups were defined, and 1466 DEGs were identified between the two subtypes. These DEGs were mainly enriched in collagen-containing extracellular matrix, cytokine-cytokine receptor interaction, the PI3K-Akt signaling pathway, and neuroactive ligand-receptor interaction. Furthermore, the ICD-low group exhibited a favorable prognosis, enhanced TTN and MUC16 mutation frequencies, increased infiltrating immune cells, and downregulated immune checkpoint expressions. Furthermore, we demonstrated that an ICD-related model (based on CD4, NLRP3, NT5E, and TLR4 genes) could forecast the prognosis of LUSC, and ICD risk scores were lower in the responder group. In summary, the predicted values of ICD-related genes (CD4, NLRP3, NT5E, and TLR4) for the prognosis and response to immunotherapy in LUSC were verified in the study, which benefits immunotherapy-based interventions for LUSC patients.

The peptide CIGB-210 inhibits HIV replication, inducing a rearrangement of vimentin intermediate filaments. The assessment of the in vitro serum and plasma stability of this peptide is important to develop an optimal pharmacological formulation. A half-life of 17.68 ± 0.59 min was calculated for CIGB-210 in human serum by reverse-phase high-performance liquid chromatography (HPLC) and mass spectrometry (MS). Eight metabolites of CIGB-210 were identified with this methodology, all of them lacking the N-terminal moiety. A previously developed CIGB-210 in-house competitive ELISA was used to compare the stability of CIGB-210 derivatives containing either D-amino acids, acetylation at the N-terminus, or both modifications. The half-life of CIGB-210 in serum was five times higher when measured by ELISA than by HPLC/MS, and twice higher in plasma as compared to serum. The substitution of D-asparagine on position 6 doubled the half-life, while D-amino acids on positions 8 and 9 did not improve the stability. The acetylation of the N-terminus resulted in a 24-fold more stable peptide in plasma. The positive effect of N-terminal acetylation on CIGB-210 serum stability was confirmed by the HPLC/MS method, as the half-life of the peptide was not reached after 2 h of incubation, which represents more than a 6.8-fold increase in the half-life with respect to the original peptide.

The emergence or reemergence of monkeypox (Mpox) and Ebola virus (EBOV) agents causing zoonotic diseases remains a huge threat to human health. Our study aimed at designing a multi-epitope vaccine (MEV) candidate to target both the Mpox and EBOV agents using immunoinformatics tools. Viral protein sequences were retrieved, and potential nonallergenic, nontoxic, and antigenic epitopes were obtained. Next, cytotoxic and helper T-cell (CTL and HTL, respectively) and B-cell (BCL) epitopes were predicted, and those potential epitopes were fused utilizing proper linkers. The in silico cloning and expression processes were implemented using Escherichia coli K12. The immune responses were prognosticated using the C-ImmSim server. The MEV construct (29.53 kDa) included four BCL, two CTL, and four HTL epitopes and adjuvant. The MEV traits were pertinent in terms of antigenicity, non-allergenicity, nontoxicity, physicochemical characters, and stability. The MEV candidate was also highly expressed in E. coli K12. The strong affinity of MEV-TLR3 was confirmed using molecular docking and molecular dynamics simulation analyses. Immune simulation analyses unraveled durable activation and responses of cellular and humoral arms alongside innate immune responses. The designed MEV candidate demonstrated appropriate traits and was promising in the prediction of immune responses against both Mpox and EBOV agents. Further experimental assessments of the MEV are required to verify its efficacy.

The objective of this work was to optimize the application of an enzymatic blend produced by Aspergillus niger ATCC 1004 on the Pimenta dioica fruits for essential oil extraction. The enzyme blend was obtained from the fermentation of cocoa bean shells, an agro-industrial residue. The effects of the enzymatic pre-treatment on the extraction yield, the chemical composition of the oil through gas chromatography, and the fruit structure through scanning electron microscopy (SEM) were assessed. A Doehlert design was used to optimize the process conditions, resulting in an extraction with 117 mL of enzyme during 77 min, which increased the extraction yield by 387.5%. The chemical composition was not altered, which proves that the enzyme blend preserves the quality of the essential oil extracted. The content of eugenol (70%), the major compound in the P. dioica essential oil, had a great increase in its concentration (560%). The enzyme activity analyses showed the presence of endoglucanase (0.4 U/mL), exoglucanase (0.25 U/mL), β-glucosidase (0.19 U/mL), and invertase (135.08 U/mL). The microscopy analyses revealed changes in the morphology of fruit surface due to the enzymatic action. These results demonstrate the great potential of using enzyme blends produced by filamentous fungi from agro-industrial residues for the essential oils extraction of interest for the pharmaceutical and food industries.

Neuropathic pain (NP) significantly impacts the quality of life due to its prolonged duration and lack of effective treatment. Recent findings suggest that targeting neuroinflammation is a promising approach for treating NP. G protein-coupled receptor 55 (GPR55), a member of the GPCR family, plays an important role in neuroinflammatory regulation. CID16020046, a GPR55 agonist, possesses promising anti-neuroinflammatory effects. Herein, the therapeutic effect of CID16020046 on NP was investigated in an NP rat model. The NP model was established using the unilateral sciatic nerve chronic constriction injury (CCI) assay. Both sham and CCI rats were intraperitoneally administered with 20 mg/kg CID16020046. NP was assessed using paw withdrawal threshold (PWT) and paw withdrawal latency (PWL). First, we showed that GPR55 was downregulated in the spinal dorsal horn of CCI rats. After CCI rats were treated with CID16020046, the values of PWT and PWL were increased, indicating their effect on pain relief. The treated rats had attenuated release of inflammatory cytokines in the spinal cord, decreased spinal malondialdehyde (MDA) levels, and increased spinal glutathione peroxidase (GSH-PX) activity. Additionally, the increased levels of phosphorylated nuclear factor (NF)-κB p65 in CCI rats were significantly alleviated by CID16020046 treatment. Mechanistically, we showed that CID16020046 significantly suppressed the activation of the Janus kinase (JAK2)/signal transducer and activator of transcription 3 (JAK2/STAT3) pathway in the spinal cord of CCI-treated rats. However, Colivelin TFA (a STAT3 agonist) abolished the effect of CID16020046 on JAK2/STAT3 activation. In conclusion, our data demonstrate that the activation of GPR55 by CID16020046 alleviates NP and neuroinflammation in CCI rats by mediating the JAK2/STAT3 pathway.

Loss of osteogenic differentiation potential of osteoblasts has been associated with the pathogenesis of osteoporosis. Thus, stimulation of osteoblastic differentiation is a therapeutic strategy for osteoporosis. Relaxin-2 is a peptide hormone with potent biological functions. However, the effects of Relaxin-2 in osteoblastic differentiation and osteoporosis have not been reported before. Here, we report a novel physiological role of Relaxin-2 in promoting osteoblastic differentiation and mineralization of MC3T3-E1 cells. Our results indicate that exposure to Relaxin-2 upregulated the expression, and elevated the activity of alkaline phosphatase (ALP) when MC3T3-E1 cells were cultured in osteogenic differentiation medium (OM). Additionally, Relaxin-2 upregulated the mRNA levels of osteocalcin (ocn), osteopontin (opn), and collagen type I alpha 1 (Col1a1). The alizarin red S staining assay revealed that Relaxin-2 promoted the mineralization of MC3T3-E1 cells. We also found that Relaxin-2 increased the expression of Runx-2 as well as the epidermal growth factor (EGF) and epidermal growth factor receptor (EGFR). Importantly, silencing of EGF abolished the effects of Relaxin-2 in osteoblastic differentiation and related gene expression. These findings suggest that Relaxin-2 stimulates osteogenic differentiation through activating EGF/EGFR signaling.