Biotechnology and applied biochemistry最新文献

Lung cancer remains a leading cause of cancer-related mortality worldwide. Our study investigates the involvement of the PD-L1/MALAT1/miR-200a-3p axis in lung tumor progression using a murine model of lung carcinogenesis. Lung tumors were induced in rats, which were divided into groups and sacrificed at different stages of tumor development. A histopathological examination was performed to assess tumor progression. Immunohistochemistry was applied to evaluate the expression of Ki-67 and programmed death-ligand 1 (PD-L1). The level of carcinoembryonic antigen (CEA) and expression analysis of metastasis-associated lung adenocarcinoma transcript 1 (MALAT1), miR-200a-3p, and zinc finger E-box binding homeobox 1 (ZEB1) were evaluated for each stage of induction. Immunohistochemical analysis demonstrated a progressive upregulation of the proliferative marker Ki-67 and the immune checkpoint protein PD-L1 during the induction process, indicative of enhanced tumor proliferation and immune evasion. Additionally, CEA levels revealed a progressive increase across induction stages, with a significant increase in advanced tumor stages, highlighting its clinical relevance as a biomarker for lung cancer progression. Expression analysis revealed dynamic upregulation of MALAT1 and downregulation of miR-200a during lung tumor induction, which correlated with advanced tumor stages and elevated PD-L1 expression, suggesting that the negative correlation between MALAT1 and miR-200a is involved in the development of lung tumors. ZEB1 expression exhibited a notable increase in the advanced stages of induction, consistent with its association with aggressive lung cancer. Our findings underscore the interplay between molecular pathways involved in lung tumor development and the potential diagnostic and therapeutic implications of the PD-L1/MALAT1/miR-200a-3p axis.

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.

The PnuC gene plays a crucial role in the complex processes related to the absorption and synthesis of the nicotinamide mononucleotide (NMN) precursor. NMN, a nicotinamide adenine dinucleotide (NAD⁺) precursor, is important for cellular energy metabolism, DNA repair, and antiaging. This study focuses on elucidating the precursor absorption mechanism and the specific function of the PnuC gene in encoding membrane transport proteins, as well as its impact on the regulation and dynamics of NMN within the cell. This understanding aims to provide insights into its potential effects on metabolic balance, illustrated through two NAD⁺ biosynthesis pathways based on renewable and readily available cytoplasmic resources, assessing the potential of PnuC gene expression in clarifying complex interactions within regulation mechanisms. Enhanced expression analysis of the PnuC gene has initiated discussions on its potential applications in treating aging-related diseases and dysfunctions, contributing to cellular health maintenance.

Recent years, intensified fed-batch culture with ultra-high seeding density (uHSD-IFB) is coming to the forefront of manufacturers' choice for its enhanced productivity. However, the effects of seed cell physiological state and aeration strategies on these processes remain underexplored due to the ultra-high seeding density. Currently, the pre-production seeding inoculum (N-1) crucial for the uHSD-IFB cultures relies heavily upon case-by-case empirical experiences. To develop a rational seeding approach as a guideline, we here explored the impact of perfusion rates and cell growth states on the subsequent uHSD-IFB processes. It was found that seed cells in the exponential growth phase with high perfusion rates in the N-1 perfusion stage allowed for higher viable cell density and titer in the production stage. In particular, lower levels of reactive oxygen species, higher proportions of G1 and S phase, and higher specific cell oxygen uptake rates (OURs) were exhibited in these cells, resulting in higher cell specific growth rates and integral of viable cell concentration (IVCC) throughout the production cultures. Further investigation into the effect of aeration strategies was carried out in the benchtop bioreactors. A final yield of 4.5 g/L, an increase of nearly 110%, was achieved by a sophisticated dual sparger system compared to the other two processes with either one l-shaped or micro-sparger. These results provide a direction for the design and establishment of high-titer processes in intensified fed-batch cultures at ultra-high seeding density. Synopsis: In this work, we first explored the impact of perfusion rates and cell growth states on the subsequent uHSD-IFB processes. Further investigation into the effect of aeration strategies of intensified fed-batch process was carried out in the benchtop bioreactors. These results provide a direction for the design and establishment of high-titer processes in intensified fed-batch cultures at ultra-high seeding density.

Bacteriocin lactococcin036019 was identified and characterized from Lactococcus lactis NCU036019, which displayed significant antibacterial activity toward foodborne pathogenic bacteria Staphylococcus aureus under various conditions. However, the in situ low-level expression of lactococcin036019 severely limited its wide application in food industry. In this study, we optimized the medium ingredients and culture conditions of L. lactis NCU036019 for maximum production of lactococcin036019. The effects of different carbon sources, nitrogen sources, inorganic salts, growth factors, surfactants, and buffer salts on the production of bacteriocin were studied using antibacterial titer and diameter of inhibitory zone as evaluation indexes. Through single-factor experiments, Plackett-Burman (PB) experiment, steepest ascent experiment and response surface methodology, yeast extract, zinc sulfate, sodium acetate, mannitol, Tween-80, and dipotassium hydrogen phosphate were identified to display significant influence on the production of bacteriocin. By optimizing Man Rogosa and Sharpe (MRS) culture medium ingredients, the antibacterial activity of lactococcin036019 in the cell-free supernatant raised from 46.19 to 300.14 Au/mL, namely, 6.5 times increased. Furthermore, the culture conditions, such as inoculation amount, culture time, and culture temperature, were optimized, and this further increased the antibacterial activity to 409 Au/mL, namely, 8.8 times increased. This study investigated the effects of culture media and conditions on the production of lactococcin036019, and they were optimized for a maximum harvest of bacteriocin, and the significant increase of bacteriocin production in L. lactis NCU036019 facilitates the application of the antibacterial substance in future work.

Ulcerative colitis affects the inner lining of the large intestine, causing discomfort, pain, and digestion issues, and sometimes leading to life-threatening complications. With proper medication, symptoms and inflammation can be reduced, improving the condition. In this research, a multiwalled carbon nanotube (MWCN)-modified circular interdigitated electrode (circular-IDE) biosensor was developed to detect the ulcerative colitis biomarker lipocalin-2 and measured at 0-2 V. A dual probing strategy with aptamer and antibody on gold nanoparticles was employed for the detection of lipocalin-2. Probe immobilization was optimized on MWCN-modified circular-IDE, and saturation of 800 nM of aptamer on the GNP-antibody facilitated the identification of lipocalin-2 at concentrations as low as 1 pg/mL, with an R² value of 0.9716 [y = 2.1058x - 2.7351]. Furthermore, lipocalin-2 spiking in serum increased the current responses in correlation with the concentrations of lipocalin-2, indicating selective identification without interference. In addition, nonimmune antibody and GNP-conjugated complementary aptamer did not increase the current responses, affirming the specific detection of lipocalin-2. This MWCN-modified circular-IDE biosensor, utilizing aptamer-antibody interactions, aids in identifying the condition of ulcerative colitis.

This study evaluates the efficacy of garbage enzyme (GE) in bioremediation to reduce pollutants in sewage drains that discharge into the natural streams and rivers. Garbage enzyme is prepared with help of brown sugar, fruit, vegetable wastes, and water in the proportion 1:3:10 (by weight), which is then applied to the samples collected from various drainage sites in Jaunpur district, Uttar Pradesh, India. Different concentrations of GE (ranging from 0% to 20%) are mixed with sewage to assess pollution reduction. Different parameters, that is, pH, dissolved oxygen (DO), total suspended solids (TSS), total dissolved solids (TDS), biochemical oxygen demand (BOD), chemical oxygen demand (COD), and CD have been measured on 3rd, 5th, 7th, 10th, 15th, and 20th days. It has been observed in the study that significant percentage reduction in BOD, COD, TSS, and TDS of 80.6%, 62.9%, 74.29%, and 43.9%, respectively, while an increase in DO is 67.8% on 20th day of experiment with 20% GE addition. The result of the study reveals that there is highest reduction occurring on the 20th day with a 20% GE concentration. It has been observed that the GE possesses protease, amylase, and lipase activity. Meaning GE contains microorganisms that breakdown carbohydrates, fats, and proteins and it hydrolyzes these complex organic molecules into simpler compounds, making them more readily biodegradable and that is the reason for substantial reduction in the pollutants. Moreover, in this study it has been observed that there is a notable decrease in foul odor emanating from the samples. Time-dependent fluctuations in pH, TDS, DO, BOD, COD, and temperature are documented, with their correlations examined. This investigation underscores GE's potential in pollution mitigation, particularly in sewage systems, and offers valuable insights for sustainable environmental management practices aimed at conserving natural water bodies. Further tests are required to check the efficiency of treatment with GE dosage higher than 20%.