Maximum Activity Research Articles

Characterisation of Bambara groundnut (Vigna subterranea (L.) Verdc.) shell waste as a potential biomass for different bio-based products.

Efforts are ongoing to utilise agricultural waste to achieve a full resource use approach. Bambara groundnut is an important crop widely grown in the sub-Saharan Africa with potential future importance because of its resilience to thrive under heightened weather uncertainty and widespread droughts that have challenged food security. After harvesting, the edible nuts are separated from the shells which are discarded as waste. Therefore, this research is aimed at characterising the chemical composition and the structural properties of Bambara groundnut shells (BGS) in view of their potential application as a biomass for different bio-products. The chemical composition of BGS was found to be 42.4% cellulose, 27.8% hemicellulose, 13% lignin and 16.8% extractives. Proximate analysis showed a high amount of volatile matter (69.1%) and low moisture (4.4%). XRD analysis confirmed crystallinity of cellulose I polymer and FTIR analysis observed functional groups of lignocellulosic compounds. Thermal stability, maximum degradation temperature and activation energy were found to be 178.5°C, 305.7°C and 49.4kJ/mol, respectively. Compared to other nutshells, BGS were found to have a relatively high amount of cellulose and crystallinity that may result in biocomposites with improved mechanical properties.

Spectroscopic Analysis of Bioactive Compounds from Latex of Calotropis gigantea L. and an Evaluation of Its Biological Activities

The current research investigation aimed to screen the bioactive compounds in the latex of Calotropis gigantea L. and evaluate its antibacterial and antioxidant properties towards clinical applications. The chemical moiety and volatile compounds of the latex of C. gigantea were detected by UV–Vis spectroscopy, FT-IR, and GC–MS analysis. The antibacterial activity was assessed using wound-inducing pathogens by well diffusion method. In addition, the antioxidant properties were determined through DPPH, ABTS, and FRAP methods. The functional groups of O–H stretch, diketonic bonds, C–O, C–N, O–C bonds, and consecutive C–H bonds were observed in the latex of C. gigantea. The major bioactive compounds were 5H-3,5a-Epoxynaphth[2,1-c]oxepin, Cholesta-5-en-3-ol, 24-propylidene-, dodecane, Lup-20(29)-Ene-3,28-Diol, (3.Beta)-, Veridiflorol, and Lanosta-8,24-dien-3-ol, acetatate, (3.beta.). Oxazole derivatives were found in the latex of C. gigantea, proved by GC–MS analysis. The aqueous-mixed latex exhibited maximum antioxidant activity as compared to methanol-mixed latex. Aqueous-mixed latex and methanol-mixed latex inhibited the growth of K. pneumoniae, P. mirabilis, S. pyogenes, Micrococcus spp., S. aureus, P. aeruginosa, and E. coli. The present study clearly reveals that latex of C. gigantea has rich bioactive compounds with significant biological activities, and can be employed to produce a novel herbal formulation against wound-inducing pathogens.

Potency of Cocoa Honey, Byproduct of Cocoa Bean Processing as Probiotic Media: Optimization of Saccharomyces boulardii Growth and Its Antimicrobial Activity

Cocoa honey, a byproduct of cocoa bean processing, contains high sugars and minerals; thus, it has potential as a fermentation medium. This research investigated the potency of cocoa honey as probiotic media for promising yeast probiotic Saccharomyces boulardii. The growth of S. boulardii in the cocoa honey medium was optimized through response surface methodology employing a central composite rotatable design to obtain maximum cell production and antimicrobial activity. The effects of inoculum size (2–5%), sucrose addition (2.0–3.0%), and fermentation time (5–20 h) were evaluated. The agar diffusion technique assessed the cocoa honey medium's antibacterial activity. Results showed that these independent variables significantly affected the production of S. boulardii cells and their microbial activity against Escherichia coli. The optimal values generated by the model with a desirability value of 0.908 were 20 h of fermentation time, 2% of sucrose addition, and 2% of inoculum size. The maximum yeast count (log10 8.07 cells/mL) and antibacterial activity (4.88 mm) were reached under these optimized conditions, which agreed with the predicted value of the RSM model. This study provided valuable information on the potency of cocoa honey as a fermentative medium for applying S boulardii to develop a probiotic drink.

Abstract Tu118: The Mitochondrial LonP1 Is Indispensable For Cardiac Maturation And Function

Background: LonP1 is a major mitochondrial ATP-dependent protease critical for mitochondrial protein quality control, However its role in cardiac function is unclear. Hypothesis: We hypothesize that LonP1 is essential for postnatal cardiac maturation and function. Methods: We employed cardiac-specific LonP1 knock-out (LonP1cKO) and Cre-control mice, assessing their heart histology (HE, trichrome), cardiac function via echocardiography, mitochondrial function by measuring oxygen consumption rate (OCR), mitochondrial complexes (I-V) activity by (ETC complex maximal activity), Iron-Sulfur (Fe-S) clusters and free radical levels by electron paramagnetic resonance (EPR) spectroscopy. The significant difference between the groups was established at p<0.05 using the Student's 't' test. Results: We found that LonP1cKO mice were not embryonically lethal; however, they exhibited an early mortality rate, died at an average of 21 days after birth with significantly lower body weight (7.02±0.16 gm vs. control (11.41±0.25 gm), p<0.0001, n=24) and increased heart weight to the tibia length ratio (0.080±0.002 gm/cm vs. control (0.047±0.0014 gm/cm), p<0.001, n=24). Echocardiography analysis showed that LonP1ckO mice had severely decreased left ventricular ejection fraction (%EF) (14.06±1.82% vs. control (70±2.9%), p<0.0001, n=16) and fractional shortening (% FS) (6.05±0.8% vs. control (39.15±2.5%), p<0.0001, n=16). Histological analysis revealed severe ventricular dilation and increased cardiac fibrosis. Transmission electron microscope analysis showed a significant loss of cristae and accumulation of electron-dense aggregation within mitochondria of LonP1cKO but not in control mice. In addition, LonP1cKO hearts showed reduced Fe-S clusters level (10.70±0.85 a.u vs. 21.27±1.91, p<0.0001, n=6), complex I-V activities (p<0.001), and electron flow (152.2±4.04 nm/min/mg vs. 265.3±23.8, p<0.001, n=6) compared to control. Conclusion: Our findings indicate that the deletion of LonP1 makes the heart unable to transition from glycolysis to substrate oxidation during the early postnatal maturation process, leading to dilated cardiomyopathy and death. This highlights the pivotal role of LonP1 in both cardiac maturation and function.

Date nawah powder as a promising waste for β-mannanase production from a new isolate Aspergillus niger MSSFW, statistically improving production and enzymatic characterization

β-Mannanase producing fungus was isolated from coffee powder waste and identified as Aspergillus niger MSSFW (Gen Bank accession number OR668928). Dates nawah powder as industrial and agricultural waste was the most inducer of β-mannanase production. The Plackett–Burman and Central Composite designs were used to improve β-mannanase titer. Optimization studies enhanced the enzyme yield with approximate 13.50-times. β-Mannanase was purified by Sephadex G-150 gel filtration column and the molecular weight was estimated to be 60 kDa by SDS–PAGE. Crude and purified β-mannanase displayed maximum activity at temperature 60 °C and 50 °C, respectively. Crude β-mannanase showed an activation energy value 2.35-times higher than the purified enzyme. Activation energy for thermal denaturation of the purified β-mannanase was 1.08-times higher than that of the crude enzyme. Purified β-mannanase exhibited higher deactivation rate constant (Kd) and lower half-life (t0.5) and decimal reduction time (D-value) compared with the crude enzyme. Thermodynamic parameters of enthalpy, entropy, and free energy values for crude and purified β-mannanase were calculated. Substrate kinetic parameters suggested that the purified β-mannanase had a strong affinity toward locust bean gum by showing 3.44-times lower Km and 1.99-times higher Vmax compared to the crude enzyme.

Potential of bacteriophage phT4A as a biocontrol agent against Escherichia coli in food matrices

Escherichia coli is one of the most prevalent foodborne pathogens, frequently found in meat and dairy products. Current decontamination methods are often associated with changes in organoleptic characteristics, nutrient loss, and potentially harmful side effects. Furthermore, despite the array of available methods, foodborne outbreaks still frequently occur. For this reason, bacteriophages (or simply phages) emerged as a natural alternative for the biocontrol of bacterial contamination in food without altering their organoleptic properties. In this study, the potential of phage phT4A was assessed in the biocontrol of E. coli in liquid (milk) and solid (ham) food matrices. Firstly, as foods have different pH and temperature values, the influence of these parameters on phage phT4A viability was also assessed to develop an effective protocol. Phage phT4A proved to be stable for long storage periods at pH 7–8 (56 days) and temperatures of 4–37 °C (21 days). Before application of phages to inactivate pathogenic bacteria in food, previous assays were carried out in Tryptic Soy Broth (TSB) to study the dynamics of phage-bacteria interaction. Then, the antibacterial potential of phage phT4A was evaluated in the two food matrices at different temperatures (4, 10 and 25 °C). This phage was more efficient at 25 °C in all tested matrices (maximum inactivation of 6.6, 3.9 and 1.8 log CFU/mL in TSB, milk and ham, respectively) than at 10 °C (maximum decrease of 4.7, 2.1 and 1.0 log CFU/mL in TSB, milk and ham, respectively) and 4 °C (maximum reduction of 2.6 and 0.7 log CFU/mL in TSB and milk, respectively). However, the decrease of temperature from 25 °C to 10 and 4 °C prevented bacterial regrowth. The results suggest that during phage treatment, a balance between an incubation temperature that provide effective results in terms of bacterial inactivation by the phages and at the same time prevents or delays bacterial regrowth, is needed. The application of phage phT4A at a temperature of 10 °C can be an effective strategy in terms of bacterial inactivation, delaying bacterial regrowth and also reducing energy costs.

Body Composition, Vascular Health, Cardiorespiratory Fitness, Lung Function, Muscle Architecture, and Physical Activity in People with Young Onset Dementia: A Case-Control Study

BackgroundBody composition, blood pressure, estimated maximal oxygen uptake (VO2max), lung function, physical activity, muscle architecture, and endothelial function had not previously been examined in people with young onset dementia. Therefore, the study measured these variables in a young onset dementia group, compared them to age-matched controls. MethodsEstimated VO2max (via the Astrand-Rhyming test), body composition, blood pressure, lung function (via spirometry), muscle architecture (via ultrasonography) and endothelial function (via flow mediated dilation) were assessed. Physical activity was measured using ActiGraph accelerometers for 7 days. ResultsWe recruited 33 participants (16 young onset dementia, 17 controls). The young onset dementia group had shorter fascicle lengths of the vastus lateralis, were sedentary for longer over a seven-day period, and completed less moderate-vigorous physical activity than controls (p=0.028, d=0.81; large effect, p=0.029, d=0.54; moderate effect, and p=0.014, d=0.97; large effect, respectively for pairwise comparisons). Pairwise comparisons suggest no differences at the p<0.05 level between young onset dementia and controls for estimated VO2max (despite a moderate effect size [d=0.66]), height, body mass, BMI, blood pressure, light physical activity, lung function, muscle thickness, pennation angle, or endothelial function. ConclusionThis study highlights differences between people with young onset dementia and controls, underscoring the need for multicomponent exercise interventions. Future interventions should target muscle architecture, increase moderate-vigorous physical activity, and reduce sedentariness, with the goal of improving quality of life and promoting functional independence.

Facilitating secretory expression of apple seed β-glucosidase in Komagataella phaffii for the efficient preparation of salidroside.

Plant-derived β-glucosidases hold promise for glycoside biosynthesis via reverse hydrolysis because of their excellent glucose tolerance and robust stability. However, their poor heterologous expression hinders the development of large-scale production and applications. In this study, we overexpressed apple seed β-glucosidase (ASG II) in Komagataella phaffii and enhanced its production from 289 to 4322 U L-1 through expression cassette engineering and protein engineering. Upon scaling up to a 5-L high cell-density fermentation, the resultant mutant ASG IIV80A achieved a maximum protein concentration and activity in the secreted supernatant of 2.3g L-1 and 41.4 kU L-1, respectively. The preparative biosynthesis of salidroside by ASG IIV80A exhibited a high space-time yield of 33.1g L-1 d-1, which is so far the highest level by plant-derived β-glucosidase. Our work addresses the long-standing challenge of the heterologous expression of plant-derived β-glucosidase in microorganisms and presents new avenues for the efficient production of salidroside and other natural glycosides.

Effect of cadmium toxicity on growth, physiochemical parameters and antioxidant system of castor seedlings

The research was aimed to determine the potential impact of cadmium contamination on Ricinus communis. The glucose-6-phosphate dehydrogenase (G6PDH) activity in the root was highest when exposed to 0.2 mM of Cd, with an increase of 15.63 % and 14.48 % at 0 and 24 h, respectively, compared to its control. However, citrate synthase (CS) activity declined in leaves, in contrast, to root, i.e., 12.22 % at 48 h of Cd stress. Isocitrate dehydrogenase (ICDH) activity was maximum in leaves at 0.2 mM of Cd at 0 and 24 h, i.e., 12.36 % and 13.08 % respectively, and later decreased in activity was seen in roots and leaves as the Cd stress increased. Moreover, the level of malate dehydrogenase (MDH) declined in leaves as the Cd level increased, while activity increased in roots at 0.4 mM of Cd i.e., 17.21 %, 17.52 %, and 10.53 % at 0, 24, and 48 h respectively. The important metabolite, glutathione level in the roots of SKP 84 was higher than in the leaf extract. A decline in biomass of up to 28.70 % and 30.91 % and plant length of up to 20.80 % and 26.10 % in shoot and roots, respectively, tolerance index was maximum at 0.2 mM, i.e., 98.62 % was seen. The leaves had 35.40 % catalase (CAT) activity, while the roots had 78.26 % guaiacol peroxidase (GPX) activity at 0.6 mM of Cd. At 0.2 mM of Cd, the maximum activity of ascorbate peroxidase (APX) was observed, with 67.32 % and 62.85 % activity in roots and leaves respectively. However, a reduction in the SOD activity was seen as the Cd stress increased. Increased Cd levels decreased chlorophyll but increased MDA and proline content in leaves at 0.8 mM of Cd, i.e., 82.92 % and 21.7 %, respectively. It indicated that R. communis SKP 84, a fusarium wilt resistance line, is also tolerant to Cd and can be used for phytoremediation in Cd-contaminated areas.

Hierarchical Threshold Pruning Based on Uniform Response Criterion.

Convolutional neural networks (CNNs) have been successfully applied to various fields. However, CNNs' overparameterization requires more memory and training time, making it unsuitable for some resource-constrained devices. To address this issue, filter pruning as one of the most efficient ways was proposed. In this article, we propose a feature-discrimination-based filter importance criterion, uniform response criterion (URC), as a key component of filter pruning. It converts the maximum activation responses into probabilities and then measures the importance of the filter through the distribution of these probabilities over classes. However, applying URC directly to global threshold pruning may cause some problems. The first problem is that some layers will be completely pruned under global pruning settings. The second problem is that global threshold pruning neglects that filters in different layers have different importance. To address these issues, we propose hierarchical threshold pruning (HTP) with URC. It performs a pruning step limited in a relatively redundant layer rather than comparing the filters' importance across all layers, which can avoid some important filters being pruned. The effectiveness of our method benefits from three techniques: 1) measuring filter importance by URC; 2) normalizing filter scores; and 3) conducting prune in relatively redundant layers. Extensive experiments on CIFAR-10/100 and ImageNet show that our method achieves the state-of-the-art performance on multiple benchmarks.

Airway mechanics alters generation of cough motor pattern.

Effects of sequential increase in airway resistance: no, low (5 kPa.s/l), high (24 kPa.s/l), and complete block in the inspiratory or expiratory phase of mechanically induced cough on the cough motor pattern were studied in 16 anesthetized (pentobarbital) spontaneously breathing cats (3.70±0.15 kg, 11♂, 5♀). Esophageal pressure and electromyographic activities of the diaphragm during inspiration and abdominal muscles during expiration were analyzed. No significant changes in the number of coughs occurred. Inspiratory occlusion caused a prolongation of cough inspiratory phase, cough inspiratory diaphragm activity, and all cough-related activity. Inspiratory occlusion along with high resistance increased inspiratory esophageal pressure amplitude, total cough cycle duration and the time between maximum activity of the diaphragm and abdominal muscles. High expiratory resistance and occlusion resulted in increased cough expiratory esophageal pressure amplitude, a longer active portion of cough expiration, and cough abdominal activity. Expiratory occlusion also prolonged cough expiratory phase, all cough activity, and total cough cycle. Significantly increased airway resistance and occlusion induce secondary, in addition to mechanical, changes in cough by significantly modulating the generated cough motor pattern. A certain level of resistance appears to be successfully compensated, resulting in minimal changes in coughing characteristics, including expiratory airflow and the rising time of the airflow. Afferent feedback from the respiratory tract, particularly volume feedback, represents a significant factor in modulating cough, mainly under various pathological conditions in the respiratory system.

Effect of Knee Angle, Contractile Activity, and Intensity of Force Production on Vastus Lateralis Stiffness: A Supersonic Shear Wave Elastography Pilot Study.

Supersonic shear image (SSI) ultrasound elastography provides a quantitative assessment of tissue stiffness using the velocity of shear waves. SSI's great potential has allowed researchers in fields like biomechanics and muscle physiology to study the function of complex muscle groups in different conditions. The aim of this study is to use SSI to investigate changes in the stiffness of the vastus lateralis (VL) muscle as a consequence of passive elongation, isometric contraction, and repeated muscle activity. In a single session, 15 volunteers performed a series of isometric, concentric, and eccentric contractions. SSI images were collected from the VL to assess its stiffness before and after the contractions and at various knee angles. Two-way within-subjects ANOVA was used to test the effects of muscle contraction type and knee angle on VL stiffness. Linear regression analysis was employed to assess the relationship between muscle stiffness and the intensity of isometric contractions. After maximal contractions, VL stiffness increased by approximately 10% compared to baseline values, and following maximal isometric (p < 0.01) and eccentric contractions (p < 0.05). Yet, there was no change in VL shear modulus at the end of concentric contractions. The relaxed VL shear modulus increased with knee flexion both before and after the knee extensor contractions (p < 0.001). A linear relationship between the shear modulus and the degree of isometric contraction was observed, although with notable individual variation (R2 = 0.125). Maximal contractile activity produces modest increases in relaxed muscle stiffness. The SSI-measured shear modulus increases linearly with the degree of isometric contraction.

Supramolecular catalysts: A critical review

This critical review on Supramolecular Catalysts provides a comprehensive overview of the evolution and breakthroughs in this innovative field. The review highlights the contemporary breakthroughs in adaptive supramolecular catalysts as well as metal-organic counterparts, and their possible applications. The review also discusses the challenges that need to be addressed in the development of responsive supramolecular catalysts, such as optimizing the encapsulation process to achieve maximum stability and catalytic activity. The review combines concepts from responsive materials with phenomena underpinning supramolecular chemistry to equip both reactive and metal-organic varieties to address major issues present in current synthetic chemical processes productively. The review concludes that the future of supramolecular catalysis looks bright, and we can expect to see continued growth and innovation in this exciting field. Overall, this review provides a succinct outline of the evolution and progression of supramolecular catalysis, underscoring contemporary breakthroughs in adaptive supramolecular catalysts as well as metal-organic counterparts, and their possible applications. It also highlights the challenges that need to be addressed in the development of responsive supramolecular catalysts.

A Patient-Driven Mobile Health Innovation in Cystic Fibrosis Care: Comparative Cross-Case Study.

Patient-driven innovation in health care is an emerging phenomenon with benefits for patients with chronic conditions, such as cystic fibrosis (CF). However, previous research has not examined what may facilitate or hinder the implementation of such innovations from the provider perspective. The aim of this study was to explain variations in the adoption of a patient-driven innovation among CF clinics. A comparative multiple-case study was conducted on the adoption of a patient-controlled app to support self-management and collaboration with health care professionals (HCPs). Data collection and analysis were guided by the nonadoption, abandonment, spread, scale-up, and sustainability and complexity assessment tool (NASSS-CAT) framework. Data included user activity levels of patients and qualitative interviews with staff at 9 clinics (n=8, 88.9%, in Sweden; n=1, 11.1%, in the United States). We calculated the maximum and mean percentage of active users at each clinic and performed statistical process control (SPC) analysis to explore how the user activity level changed over time. Qualitative data were subjected to content analysis and complexity analysis and used to generate process maps. All data were then triangulated in a cross-case analysis. We found no evidence of nonadoption or clear abandonment of the app. Distinct patterns of innovation adoption were discernable based on the maximum end-user activity for each clinic, which we labeled as low (16%-23%), middle (25%-47%), or high (58%-95%) adoption. SPC charts illustrated that the introduction of new app features and research-related activity had a positive influence on user activity levels. Variation in adoption was associated with providers' perceptions of care process complexity. A higher perceived complexity of the value proposition, adopter system, and organization was associated with lower adoption. In clinics that adopted the innovation early or those that relied on champions, user activity tended to plateau or decline, suggesting a negative impact on sustainability. For patient-driven innovations to be adopted and sustained in health care, understanding patient-provider interdependency and providers' perspectives on what generates value is essential.

Termite gut microbiome being a rich source of potential lignocellulolytic bacterial strains

A study was carried out to identify prospective cellulose degrading bacteria isolated from termite gut collected from various agriculture sites. Eight of the bacterial isolates displayed a very distinct zone of clearing on CMC (Carboxylmethyl cellulose) agar media after staining with 1% Congo red, indicating that they may have cellulose degrading ability. On a CMC agar plate, CRDB6 had the largest colony diameter (3.08 cm) as well as clear zone diameter (0.96 cm), however CRDB 1 has maximum cellulolytic activity (1.45 cm) followed by CRDB5 and CRDB6 (1.33 cm and 1.28 cm) respectively. CRDB3, CRDB4 and CRDB9 showed Gram negative reaction among crop residue degrading bacterial isolates, while the remainder showed Gram positive reaction. The morphological characteristics of the bacteria isolates, such as colony shape, elevation, margin, colour and opacity, differed. Isolates CRDB6 produced highest β-glucosidase activity (12.66 m L-1) followed by CRDB1 (12.20 mg m L-1) and CRDB5 (10.21 m L-1) in Luria broth culture. Indole acetic acid production (with tryptophan) was greater in CRDB1 (389.33 mg/ml), CRDB6 (356.9 m L-1) and CRDB3 (315.96 m L-1) while as P-solubilization potential was peaked in CRDB6 (2.41 m L-1) followed by CRDB4 (1.76 m L-1) and CRDB3. Overall, isolate CRDB6 exhibited the highest cellulolytic activity as compared to rest of the bacterial isolates. These results will help in preparing bacterial consortia for the management of crop residues.

Isolation and Characterization of a (Surfactin-Like Molecule) Produced by Bacillus subtilis: Antagonistic Impact on Root-Knot Nematodes

Plant-parasitic nematodes are severe soil-borne pathogens that cause significant damage to agricultural products each year, resulting in substantial financial losses globally. Thus, there is an urgent need to identify novel biological control agents or nematicides. The nematicidal potential of Bacillus subtilis-derived lipopeptides against Meloidogyne incognita was investigated at various concentrations (35 ppm, 25 ppm, 15 ppm, 5 ppm) under in vitro conditions. Egg hatching inhibition and mortality of second-stage juveniles (J2s) of M. incognita were analyzed after exposure for 6, 12, 24, 48, and 96 hours. Data showed that with the increase in concentration and exposure period, egg hatching inhibition and percent mortality increases. Maximum percent mortality of J2s was reported at 35ppm i.e., 45%, 55%, 67.75%, 77% and 85% at 6, 12, 24, 48 and 96 hrs, respectively. The maximum ovicidal activity was reported at 35ppm concentration, with 84.61% of eggs hatching inhibition on 96 hrs of the exposure period. The bacterial culture suspension of Bacillus subtilis and Pseudomonas putida at1.2x108cfu/ml, and the crude lipopeptide (35ppm) was also investigated as a biological control agent against M. incognita on tomato in a pot experiment under glasshouse condition. Combinational treatment of P. putida and B. subtilis culture, prior to inoculation of M. incognita on tomato plant caused a significant increase in plant growth attributes and in biochemical parameters over the inoculated control. In the same treatment, the maximum reduction in nematode population and root galling was recorded. However, in the crude lipopeptide experiment study, root dip and inoculation of crude lipopeptide in tomato after the introduction of M. incognita caused a major augment in all the parameters over the inoculated control. MALDI-TOF MS analysis of crude lipopeptide shows surfactin like molecules at m/z 1058 [M+Na]+. It is concluded that crude lipopeptide or combinational treatment of B. subtilis and P. putida culture suspension can be employed as a biocontrol agent against M. incognita and may act as a source of a novel nematicidal agent of bacterial origin.

TIPRL Regulates Stemness and Survival in Lung Cancer Stem Cells through CaMKK2-CaMK4-CREB Feedback Loop Activation.

Frequent recurrence and metastasis caused by cancer stem cells (CSCs) are major challenges in lung cancer treatment. Therefore, identifying and characterizing specific CSC targets are crucial for the success of prospective targeted therapies. In this study, it is found that upregulated TOR Signaling Pathway Regulator-Like (TIPRL) in lung CSCs causes sustained activation of the calcium/calmodulin-dependent protein kinase kinase 2 (CaMKK2) signaling pathway by binding to CaMKK2, thereby maintaining stemness and survival. CaMKK2-mediated activation of CaM kinase 4 (CaMK4) leads to phosphorylation of cAMP response element-binding protein (CREB) at Ser129 and Ser133, which is necessary for its maximum activation and the downstream constitutive expression of its target genes (Bcl2 and HMG20A). TIPRL depletion sensitizes lung CSCs to afatinib-induced cell death and reduces distal metastasis of lung cancer in vivo. It is determined that CREB activates the transcription of TIPRL in lung CSCs. The positive feedback loop consisting of CREB and TIPRL induces the sustained activation of the CaMKK2-CaMK4-CREB axis as a driving force and upregulates the expression of stemness- and survival-related genes, promoting tumorigenesis in patients with lung cancer. Thus, TIPRL and the CaMKK2 signaling axis may be promising targets for overcoming drug resistance and reducing metastasis in lung cancer.

Isolation and Identification of Xylanase Producing Bacteria from Soil Samples of Kerala

Microorganisms play a very vital role in the biological recycling of plant bio-matter. Xylan is the second most plentiful polysaccharide and is found as an interface between lignin and other polysaccharide in the secondary cell wall. When these polysaccharides are hydrolyzed by a suitable strain of bacteria it will become an end to many environmental issues and had many useful applications such as in paper and pulp industry, food and feed industry, bread quality improvement, agro waste treatment, seed germination, generation of bio-fuels etc. To isolate a suitable strain of bacteria which possess good xylanase activity several samples of soils were collected from various regions of Kerala and were undergone a three step enrichment process. In the primary screening 85 bacterial strains were obtained. 22 isolates produces clear zone during plate assay using beech wood xylan. The isolates which possess more than 0.5cm clear zone were undergone final screening process and the strain PSS 1 showed the maximum enzyme activity of 112.38 U/ml.

Sequential cultivation method for β-fructofuranosidase production from Aspergillus tamarii URM4634, evaluation of their biochemical and kinetic/thermodynamic characteristics, and application on sucrose hydrolysis.

The present study focused on evaluating the sequential fermentation (SF) method for FFase production from Aspergillus tamarii URM4634 using soybean bran as substrate. The SF was performed using soybean bran as substrate at 72h and 30°C and the maximum hydrolytic activity (44.00 U mL-1), corresponding to an increase of 2.98-fold to about SmF using sucrose as substrate. Already the maximum transfructosylating activity was 26.10 U mL-1. The FFase presents maximum hydrolytic activity at pH 5.0-6.0 and transfructosylating at pH 6.0 and 60°C for both enzyme activities. The enzyme showed a typical hydrolytic kinetic profile evidenced by more affinity by sucrose hydrolysis reaction than the fructosyl transfer one. From kinetic and thermodynamic data of thermal denaturation, it was observed that the enzyme presents suitable at 55°C, evidenced by the large half-life (990.21min) and D values (3289.41min). The maximum release of reducing sugars (8.45g L-1) was obtained in hydrolysis of 20% sucrose during 180min. The results obtained for FFase production by SF proved that this method can be used satisfactorily for sucrose-degrading enzymes and can contribute to the development of the SF technique to produce different industrial-interest enzymes.

Efficient heterologous expression of cellobiose 2-epimerase gene in Escherichia coli under the control of T7 lac promoter without addition of IPTG and lactose

In this study, the cellobiose 2-epimerase gene csce from Caldicellulosiruptor saccharolyticus was expressed in Escherichia coli using TB medium containing yeast extract Oxoid and tryptone Oxoid. Interesting, it was found that when the concentration of isopropyl-beta-d-thiogalactopyranoside (IPTG) and lactose was 0 (no addition), the activity of cellobiose 2-epimerase reached 5.88 U/mL. It was 3.70-fold higher than the activity observed when 1.0 mM IPTG was added. When using M9 medium without yeast extract Oxoid and tryptone Oxoid, cellobiose 2-epimerase gene could not be expressed without IPTG and lactose. However, cellobiose 2-epimerase gene could be expressed when yeast extract Oxoid or tryptone Oxoid was added, indicating that these supplements contained inducers for gene expression. In the absence of IPTG and lactose, the addition of soy peptone Angel-1 or yeast extract Angel-1 to M9 medium significantly upregulated the expression of cellobiose 2-epimerase gene in E. coli BL21 pET28a-csce, and these inductions led to higher expression levels compared to tryptone Oxoid or yeast extract Oxoid. The relative transcription level of csce was consistent with its expression level in E. coli BL21 pET28a-csce. In the medium TB without IPTG and lactose and containing yeast extract Angel-1 and soy peptone Angel-1, the activity of cellobiose 2-epimerase reached 6.88 U/mL, representing a 2.2-fold increase compared to previously reported maximum activity in E. coli. The significance of this study lies in its implications for efficient heterologous expression of recombinant enzyme proteins in E. coli without the need for IPTG and lactose addition.