Effective Concentration Range Research Articles

Virulence, structure, and triadimefon sensitivity of the Puccinia striiformis f. sp. tritici population in Shaanxi Province, China.

Stripe rust, caused by Puccinia striiformis f. sp. tritici (Pst), is the most destructive fungal disease affecting wheat in China, especially in Shaanxi Province, an important epidemiological region connecting the western Pst over-summer regions and the central and eastern spring epidemic regions in the country. In the present study, 291 Pst isolates from Shaanxi Province were studied for their virulence using two sets of wheat differentials, population structure using single-nucleotide polymorphism (SNP) markers, and sensitivity to fungicide. When the isolates were tested on the Chinese differentials of 19 wheat cultivars, 72 races were identified, which belonged to three groups, including the Guinong 22 group (48.45%), Hybrid 46 group (31.62%), and Suwon 11 group (19.93%). The three most predominant races were CYR34 (15.46%), G22-14 (11.68%), and CYR32 (10.65%). When the isolates were tested on the 18 Yr single-gene differentials, 95 races were identified, but none of the isolates were virulent to either Yr5 or Yr15. Cluster analyses of the virulence data based on the two sets of differentials and the SNP marker data consistently separated the Shaanxi Pst population into two clusters in the central part and southern part of the Province. Triadimefon sensitivity testing across different concentrations showed a broad range of half-maximal effective concentration (EC50) values, from 0.03 to 5.99 μg mL-1, with a mean EC50 of 0.46 μg mL-1. The majority of isolates (90.72%) were sensitive to the fungicide. The correlation analyses of the virulence, SNP marker, and the triadimefon sensitivity data showed no significant correlations, except a logarithmic relationship between the EC50 value and the number of avirulence factors. This study is the first to determine the relationship of virulence and SNP markers with triadimefon sensitivity in a regional Pst population.The findings provide valuable insights for breeding resistant wheat cultivars and integrated management of stripe rust.

Prediction of teicoplanin plasma concentration in critically ill patients: a combination of machine learning and population pharmacokinetics.

Teicoplanin has been widely used in patients with infections caused by Staphylococcus aureus, especially for critically ill patients. The pharmacokinetics (PK) of teicoplanin vary between individuals and within the same individual. We aim to establish a prediction model via a combination of machine learning and population PK (PPK) to support personalized medication decisions for critically ill patients. A retrospective study was performed incorporating 33 variables, including PPK parameters (clearance and volume of distribution). Multiple algorithms and Shapley additive explanations were employed for feature selection of variables to determine the strongest driving factors. The performance of each algorithm with PPK parameters was superior to that without PPK parameters. The composition of support vector regression, categorical boosting and a backpropagation neural network (7:2:1) with the highest R2 (0.809) was determined as the final ensemble model. The model included 15 variables after feature selection, of which the predictive performance was superior to that of models considering all variables or using only PPK. The R2, mean absolute error, mean squared error, absolute accuracy (±5 mg/L) and relative accuracy (±30%) of external validation were 0.649, 3.913, 28.347, 76.12% and 76.12%, respectively. Our study offers a non-invasive, fast and cost-effective prediction model of teicoplanin plasma concentration in critically ill patients. The model serves as a fundamental tool for clinicians to determine the effective plasma concentration range of teicoplanin and formulate individualized dosing regimens accordingly.

Plasma, brain and spinal cord concentrations of caffeine are reduced in the SOD1G93A mouse model of amyotrophic lateral sclerosis following oral administration

Modifications to the small intestine and liver are known to occur during the symptomatic disease period of amyotrophic lateral sclerosis (ALS), a member of the motor neuron disease (MND) family of neurodegenerative disorders. How these modifications impact on oral absorption and pharmacokinetics of drugs remains unknown. In this study, model drugs representing different mechanisms of intestinal transport (caffeine for passive diffusion, digoxin for P-glycoprotein efflux, and sulfasalazine for breast cancer resistance protein efflux) were administered via oral gavage to postnatal day 114–120 male and female SOD1G93A mice (model of familial ALS) and wild-type (WT) littermates. Samples of blood, brain and spinal cord were taken at either 15, 30, 60 or 180 min after administration. In addition, the in vivo gastric emptying of 70 kDa fluorescein isothiocyanate-dextran (FITC-dextran) and the ex vivo intestinal permeability of caffeine were assessed. The area under the plasma concentration–time curves (AUCplasma) of digoxin and sulfasalazine were not significantly different between SOD1G93A and WT mice for both sexes. However, the AUCplasma of caffeine was significantly lower (female: 0.79-fold, male: 0.76-fold) in SOD1G93A compared to WT mice, which was associated with lower AUCbrain (female: 0.76-fold, male: 0.80-fold) and AUCspinal cord (female: 0.81-fold, male: 0.82-fold). The AUCstomach of caffeine was significantly higher (female: 1.5-fold, male: 1.9-fold) in SOD1G93A compared to WT mice, suggesting reduced gastric emptying in SOD1G93A mice. In addition, there was a significant reduction in gastric emptying of FITC-dextran (0.66-fold) and ex vivo intestinal permeability of caffeine (0.52-fold) in male SOD1G93A compared to WT mice. Reduced systemic and brain/spinal cord exposure of caffeine in SOD1G93A mice may therefore result from alterations to gastric emptying and small intestinal permeability. Specific dosing requirements may therefore be required for certain medicines in ALS to ensure that they remain in a safe and effective concentration range.

Harnessing the Potential of Harpin Proteins: Elicitation Strategies for Enhanced Secondary Metabolite Accumulation in Grapevine Callus Cultures

Grapes and grape products are rich in secondary metabolites such as phenolic compounds and anthocyanins, which have antioxidant properties. These compounds possess health-promoting attributes, including cardioprotective, antimicrobial, and anticancer effects. In recent years, biotechnological methods have been employed to produce high quantities and purity of secondary metabolites under in vitro conditions, aiming to elucidate their complex functions and optimize production methods. However, the potential effects of harpin proteins on the accumulation of secondary compounds in callus cultures have not been investigated thus far. Harpin proteins, encoded by the hrp gene clusters in Gram-negative phytopathogens, are known to trigger defense responses in various plant species by promoting the accumulation of secondary compounds. These findings suggest that harpin proteins may have the potential to enhance secondary metabolite accumulation in callus cultures. This study therefore investigated the potential of applying different concentrations of harpin protein (0, 0.1, 1, 10, and 100 ppm) to increase secondary metabolite production in calluses derived from petioles of the “Horoz Karası” grape cultivar. Our findings revealed that 1 and 10 ppm harpin treatments resulted in the highest anthocyanin accumulations, with 17.21 and 16.57 CV/g, respectively, representing 1.95- and 1.87-fold increases compared to control treatments, respectively. Total phenolic content peaked at 0.39 mg GAE g−1 FW with the 1 ppm harpin treatment, representing a 4.33-fold increase over the control. Total flavonol levels reached their highest levels at 0.027 mg CE g−1 FW with 1 and 10 ppm harpin concentrations, resulting in a 2.25-fold increase compared to the control. The highest averages for total flavonol content were recorded at 0.024 and 0.021 mg RE g−1 FW with 1 and 10 ppm harpin concentrations, respectively, representing 1.5- and 1.3-fold increases over the control. Principal component analysis (PCA) corroborated the results obtained from the heatmap analysis, indicating that harpin applications at 1 and 10 ppm were the most effective concentration range for maximizing secondary metabolite synthesis, while very low or high concentrations diminished these effects. These findings offered valuable insights for optimizing the production of high-value bioactive compounds, which can be utilized in various fields such as medicine, pharmaceuticals, food, and cosmetics. These results are expected to serve as a valuable reference for elucidating the mechanisms by which harpin proteins, rarely used in vitro, exert their effects on grapevine calluses, contributing to the literature in this domain.

Understanding the fatigue behaviour of Ti–6Al–4V manufactured via various additive processes

Additive Manufacturing (AM) is receiving widespread attention from both industry and academia who are looking to benefit from the numerous advantageous possibilities that AM processes have to offer, such as the potential to design and produce highly complex bespoke geometries with minimal material wastage. Yet, despite this, AM also has some drawbacks. Some of the most significant include the presence of process-induced defects and the inherent surface roughness of an AM built component, both of which can have a considerable influence on the mechanical properties of the final product. This research will investigate the role of an as-built surface on the fatigue properties of AM Ti–6Al–4V manufactured by electron beam melting (EBM), laser powder bed fusion (L-PBF) and laser metal deposition with wire (LMD-w). Fatigue results have been generated alongside advanced surface profilometry, microstructural, defect and fractographic analyses that have revealed that whilst the surface roughness in the majority of instances is the primary factor impacting the fatigue performance on AM material, it cannot be considered alone. It was found that the inherent as-built (AB) surface finish was significantly different across the various AM processes, inducing a range of effective stress concentrations and thus, a contrasting impact on the resulting fatigue performance. Results from each variant have been compared against a machined and polished equivalent, to provide a further consideration as to whether the as-built surface would be suffice from a time and economical viewpoint. Statistical analysis of the generated results also allowed for an extrapolation of predicted fatigue lives in the very high cycle regime for the alternative AM Ti–6Al–4V variants.

Evaluation of Short-Term Mussel Test for Estimating Toxicity.

Effect concentrations of ammonia, nickel, sodium chloride, and potassium chloride from short-term 7-day tests were compared to those from standard chronic 28-day toxicity tests with juvenile mussels (fatmucket, Lampsilis siliquoidea) to evaluate the sensitivities of the 7-day tests. The effect concentrations for nickel (59 µg Ni/L), chloride (316-519 mg Cl/L, a range from multiple tests), and potassium (15 mg K/L) obtained from the 7-day tests were within a range of effect concentrations for each corresponding chemical in the 28-day tests (41-91 µg Ni/L, 251->676 mg Cl/L, 15-23 mg K/L), whereas the 7-day ammonia effect concentration (0.40 mg/L total ammonia nitrogen; TAN) was up to 3.3-fold greater than the 28-day effect concentrations (0.12-0.36 mg TAN/L) but with overlapped 95% confidence limits. These results indicate that the 7-day tests produced similar estimates compared to the 28-day tests. Further studies are needed to evaluate the 7-day test sensitivity using additional chemicals with different modes of toxic action. Environ Toxicol Chem 2024;43:2020-2025. Published 2024. This article is a U.S. Government work and is in the public domain in the USA.

Preparation of protein-stabilized Litsea cubeba essential oil nano-emulsion by ultrasonication: Bioactivity, stability, in vitro digestion, and safety evaluation

Litsea cubeba essential oil (LCEO) has garnered widespread attention due to its robust biological activity. However, challenges such as high volatility, limited water solubility, and low bioavailability impede its application. Nano-emulsion encapsulation technology offers an effective solution to these issues. In this study, we prepared litsea cubeba essential oil nano-emulsion (LCEO-NE) for the first time using whey protein (WP) as the emulsifier through an ultrasonic-assisted method, achieving high efficiency with minimal energy consumption. Transmission electron microscopy and dynamic light scattering analyses revealed that the nanoparticles were uniformly spherical, with a particle size of 183.5 ± 1.19 nm and a zeta potential of −35.5 ± 0.95 mV. Stability studies revealed that LCEO-NE exhibited excellent thermal and salt stability, maintaining its integrity for up to four weeks when stored at 4 °C and 25 °C. In vitro digestion assays confirmed the digestibility of LCEO-NE. Furthermore, evaluation of the DPPH, ABTS, and antimicrobial activities revealed that LCEO-NE displayed superior bacteriostatic and antioxidant properties compared to LCEO. Scanning electron microscopy elucidated that its bacteriostatic effect involved the disruption of bacterial microstructure. Hemocompatibility and cytotoxicity assays demonstrated the safety of LCEO-NE within the effective concentration range. This research supports the utilization of nanoparticles for encapsulating LCEO, thereby enhancing its stability and bioactivity, and consequently expanding its applications in the food and pharmaceutical industries.

Oral docetaxel plus encequidar – A pharmacokinetic model and evaluation against IV docetaxel

The development of optimized dosing regimens plays a crucial role in oncology drug development. This study focused on the population pharmacokinetic modelling and simulation of docetaxel, comparing the pharmacokinetic exposure of oral docetaxel plus encequidar (oDox + E) with the standard of care intravenous (IV) docetaxel regimen. The aim was to evaluate the feasibility of oDox + E as a potential alternative to IV docetaxel. The article demonstrates an approach which aligns with the FDA’s Project Optimus which aims to improve oncology drug development through model informed drug development (MIDD). The key question answered by this study was whether a feasible regimen of oDox + E existed. The purpose of this question was to provide an early GO / NO-GO decision point to guide drug development and improve development efficiency. Methods: A stepwise approach was employed to develop a population pharmacokinetic model for total and unbound docetaxel plasma concentrations after IV docetaxel and oDox + E administration. Simulations were performed from the final model to assess the probability of target attainment (PTA) for different oDox + E dose regimens (including multiple dose regimens) in relation to IV docetaxel using AUC over effective concentration (AUCOEC) metric across a range of effective concentrations (EC). A Go / No-Go framework was defined—the first part of the framework assessed whether a feasible oDox + E regimen existed (i.e., a PTA ≥ 80%), and the second part defined the conditions to proceed with a Go decision. Results: The overall population pharmacokinetic model consisted of a 3-compartment model with linear elimination, constant bioavailability, constant binding mechanics, and a combined error model. Simulations revealed that single dose oDox + E regimens did not achieve a PTA greater than 80%. However, two- and three-dose regimens at 600 mg achieved PTAs exceeding 80% for certain EC levels. Conclusion: The study demonstrates the benefits of MIDD using oDox + E as a motivating example. A population pharmacokinetic model was developed for the total and unbound concentration in plasma of docetaxel after administration of IV docetaxel and oDox + E. The model was used to simulate oDox + E dose regimens which were compared to the current standard of care IV docetaxel regimen. A GO / NO-GO framework was applied to determine whether oDox + E should progress to the next phase of drug development and whether any conditions should apply. A two or three-dose regimen of oDox + E at 600 mg was able to achieve non-inferior pharmacokinetic exposure to current standard of care IV docetaxel in simulations. A Conditional GO decision was made based on this result and further quantification of the “effective concentration” would improve the ability to optimise the dose regimen.

Enhanced osteogenic and antibacterial properties of titanium implant surface modified with Zn-incorporated nanowires: Preclinical invitro and invivo investigations.

This study aimed to synthesize zinc-incorporated nanowires structure modified titanium implant surface (Zn-NW-Ti) and explore its superior osteogenic and antibacterial properties invitro and invivo. Zn-NW-Ti was synthesized via displacement reactions between zinc sulfate solutions and the titanium (Ti) surface, which was pretreated by hydrofluoric acid etching and hyperthermal alkalinization. The physicochemical properties of the Zn-NW-Ti surface were examined. Moreover, the biological effects of Zn-NW-Ti on MC3T3-E1 cells and its antibacterial property against oral pathogenic bacteria (Staphylococcus aureus, Porphyromonas gingivalis, and Actinobacillus actinomycetemcomitans) compared with sandblasted and acid-etched Ti (SLA-Ti) and nanowires modified Ti (NW-Ti) surface were assessed. Zn-NW-Ti and SLA-Ti modified implants were inserted into the anterior extraction socket of the rabbit mandible with or without exposure to the mixed bacterial solution (S. aureus, P. gingivalis, and A. actinomycetemcomitans) to investigate the osteointegration and antibacterial performance via radiographic and histomorphometric analysis. The Zn-NW-Ti surface was successfully prepared. The resultant titanium surface appeared as a nanowires structure with hydrophilicity, from which zinc ions were released in an effective concentration range. The Zn-NW-Ti surface performed better in facilitating the adhesion, proliferation, and differentiation of MC3T3-E1 cells while inhibiting the colonization of bacteria compared with SLA-Ti and NW-Ti surface. The Zn-NW-Ti implant exhibited enhanced osseointegration invivo, which was attributed to increased osteogenic activity and reduced bacterial-induced inflammation compared with the SLA-Ti implant. The Zn-incorporated nanowires structure modified titanium implant surface exhibited improvements in osteogenic and antibacterial properties, which optimized osteointegration in comparison with SLA titanium implant surface.

A Flexible Sensor for Vancomycin Fabricated By Depositing Molecularly Imprinted Carbon Paste on a Wire

1. Introduction Vancomycin (VCM), an antibacterial drug, has a narrow effective blood concentration range, requiring therapeutic drug monitoring (TDM) to measure blood levels and optimize dosing regimens . Neonates, in particular, have large individual differences in the rate of drug metabolism, so strict TDM is necessary, but the dilemma is that the amount of blood drawn must be kept to a minimum. Therefore, we decided to develop a method of TDM without blood sampling by implanting a sensor that detects VCM under the skin. The sensor must be flexible enough not to damage the subcutaneous tissue. We have developed a sensor using a MIP carbon paste (MIP-CP) electrode, in which graphite particles with molecularly imprinted polymers are mixed with oil. Since higher mechanical strength is required for indwelling sensors, we fabricated MIP-carbon dough (MIP-CD) electrodes by mixing MIP with a thermosetting silicon resin prepolymer and thermosetting it. This was applied to a wire to develop a flexible wire-type sensor, and its performance was evaluated. 2. 2. experimental method 2-1. MIP graft polymerization /MIP-CD fabricationMethacrylic acid, ethylene glycol dimethacrylate, and allylamine carboxypropionic-3-ferrocene were graft copolymerized on the surface of graphite particles with fixed polymerization initiator in the presence of template VCM. Then, after washing and drying with sodium chloride solution and distilled water, the uncured MIP-CD was mixed with thermosetting silicone resin at a weight ratio of 7:3. Non-imprinted polymer (NIP)-CD, which was prepared by the same procedure without adding a template during synthesis, was also prepared for comparison.2-2. Fabrication of MIP-CD sensor Carbon ink was applied to a 20 mm tip of a 50 mm tin-plated copper wire, and uncured MIP-CD was applied on top. Then, silicon resin was applied for insulation to limit the effective area of the MIP-CD electrode, and both the insulation and MIP-CD were heat-cured in a dryer to fabricate the wire electrode shown in Fig. 1.2-3. measurement methodDifferential pulse voltammetry was performed on the MIP (or NIP)-CD sensor in phosphate buffer containing VCM or teicoplanin (TEIC), which has a similar structure, and the relationship between current and concentration was plotted. 3. Results and Discussion A comparison of the sensitivity of MIP-CD and NIP-CD electrodes to VCM is shown in Fig. 2: the current at MIP-CD increased linearly with increasing VCM concentration, while that at NIP-CD was insensitive. This result indicates that the current at the MIP-CD electrode reflects the specific binding of VCM to the imprinted binding site. As shown in Fig. 3, the MIP-CD electrode was not sensitive to TEIC and showed high selectivity. Conclusion The electrode with MIP-CD fixed on a wire is strong and flexible, yet has a specific response to VCM, and is expected to be used as an indwelling subcutaneous sensor.*This work was supported by the Bridging Research Program of the Japan Agency for Medical Research and Development (Keio University Center). Figure 1

HPLC-UV profiles of Cynanchum auricutalaum, Cynanchum bungei and Cynanchum wilfordii and relationships of their antioxidant activities

The method for simultaneous determination of the two types of principal components, C21- steroids and acetophenones, in Cynanchum bungei, C. auriculatum and C. wilfordii by high-performance liquid chromatography-ultraviolet (HPLC-UV) was developed for the first time. Under the optimized conditions, good linearities (R2≥0.999) were obtained for all analytes, and relative standard deviations (RSDs) of HPLC-UV method validation ranged from 0.01-1.62%. The results of DPPH (1,1-diphenyl 2-picryl hydrazyl) and ABTS (2,2-azobis-3ethylbenzthiazoline-6-sulfonic acid) free radical scavenging assays indicated that the three species had a significant antioxidant activity, with the half maximal effective concentration (EC50) range of 64.56-593.38 μg/mL. Combined with bivariate analysis, the fingerprint-activity relationship of the offline antioxidant activity of the three species with their fingerprints peak was studied, and the results revealed a dose-effect relationship between acetophenones and antioxidant activity. This study provides a theoretical basis for the correct identification and application of the antioxidant activities of C. auriculatum, C. wilfordii and C. bungei.

Fluorimetric monitoring of vancomycin using an allosteric probe-initiated sensing platform

Vancomycin is the first-line drug for infections of methicillin-resistant Staphylococcus aureus (MRSA) and multi-drug-resistant bacteria. The effective therapeutic concentration range of vancomycin is narrow, so it's essential to implement vancomycin therapeutic drug monitoring. However, conventional detection methods have disadvantages of expensive equipment, complicated operation, or poor reproducibility. Herein, a fluorescent sensing platform initiated by an allosteric probe was constructed for simple and sensitive monitoring of vancomycin at a low cost. The key point of this platform is the well-designed allosteric probe, which comprises an aptamer and a trigger sequence. When vancomycin exists, the combination of vancomycin and the aptamer will lead to a conformational change of the allosteric probe, thus exposing the trigger sequence. The trigger can react with the molecular beacon (MB) to generate fluorescent signals. In addition, the allosteric probe combined with hybridization chain reaction (HCR) was applied to develop an amplified platform, the linear range is from 0.5 μg mL−1 to 50 μg mL−1 with the limit of detection (LOD) of 0.26 μg mL−1. Most importantly, this allosteric probe-initiated sensing platform shows good detection ability in human serum samples, and it also indicates great correlation and accuracy compared with HPLC. The present simple and sensitive allosteric probe-initiated platform has the potential to support the therapeutic drug monitoring of vancomycin, which is of great significance to promote the rational use of antibiotics in clinics.

Choline chloride:glycerol deep eutectic solvents assist in the permeation of daptomycin across Caco-2 cells mimicking intestinal bilayer

Daptomycin is a cyclic lipopeptide antibiotic used for complicated skin infections and right-sided endocarditis. The oral absorption of daptomycin is limited because of its restricted permeability across the intestinal epithelium, low lipophilicity, low bioavailability due to enzymatic degradation and high molecular weight. This study investigates the enhancement in intestinal absorption of daptomycin using a new class of permeation enhancers. Caco-2 cell-based model, mimicking the intestinal bilayer, was used as an in-vitro mucosal barrier to examine the diffusion of daptomycin. The study explored the potential of deep eutectic solvents (DESs) as oral permeation enhancers, in comparison with sodium caprate, a well-known intestinal permeation enhancer. Safe and non-toxic concentrations of DESs were determined using an MTT assay. Accordingly, a safe and effective concentration range was employed for enhancing the permeation of daptomycin. Diffusion studies performed using RP-HPLC indicated an increase in permeation of daptomycin in the presence of sodium caprate and DESs. Confocal microscopy enabled elucidation of the mechanism of permeation enhancement via reduced expression of cell junction proteins. Both, sodium caprate and DESs lowered the expression of ZO-1 protein, due to the loosening of tight junctions between Caco-2 cells and thus enhanced permeation of daptomycin. Therefore, DESs can be used as safe and effective permeation enhancers for daptomycin and explored for other oral peptide-based therapeutics .

PLA Films Containing Montmorillonite Nanoclay–Citronella Essential Oil Hybrids for Potential Active Film Formulation

This work evaluated the synergistic effect of citronella essential oil (Ct) and montmorillonite (MMT) (called hybrid compound) incorporated in Poly(lactic acid) (PLA) films at different concentrations (3, 10, 15, and 20 wt%). PLA films were characterized using X-ray diffraction, SEM, TGA, and DSC considering their mechanical properties and essential oil migration. XRD analysis showed the effective interaction between MMT and oil. Thermal analysis, SEM, and mechanical tests were essential to understand the saturation point of the PLA composites. Samples with 15% and 20% of Ct showed a crystallinity reduction of 0.5% compared to samples with 3% and 10% of Ct. PLA/MMT–Ct showed a reduction in tensile strength of the order of 16 and 24, correlated to 15% and 20% of the Ct content, respectively, compared to PLA/MMT–Ct3%. Migration tests showed fast oil delivery correlated with high oil concentration, as evidenced using the PLA/MMT–Ct20% sample results, which showed an estimated release of 50% in the first 150 h due to system saturation, and the remaining being released in the last 350 h. Therefore, the migration tests provide an effective Ct concentration range promising for application with active packaging due to the intrinsic antimicrobial properties of Ct.

The antiproliferative potential and mechanism of action of metformin in MCF-7 cells.

The current study aimed to investigate the potential antiproliferative activity of metformin, the effective concentration range, and the mechanism of action. Human breast cancer cells, MCF-7 were treated with a serial dilution of metformin (10-150μM) for 24 and 48h. Potential antiproliferative activity of metformin and its ability in inducing cellular apoptosis and autophagy were also investigated. Metformin inhibited MCF-7 proliferation in a concentration and time dependent manner, with 80μM as the most effective concentration. Compared with nontreated cells, metformin induced significant levels of autophagy and apoptosis, which were confirmed by the reduction of mTOR and BCL-2 protein expression. The study confirms the antiproliferative activity of metformin, which may likely occur through AMPK signaling pathway.

The study of structure and effects of two new proanthocyanidins from Anogeissus pendula leaves on rumen enzyme activities.

Two novel proanthocyanidins, (2R, 3R)-(+)-Gallocatechin-(4β → 8)4-(2R, 3R)-(+)-gallocatechin (compound 1) and 3-O-galloyl-(2S, 3S)-(-)-epicatechin-(4α → 8)-[3-O-galloyl-(2S, 3S)-(-)-epicatechin (4α → 8)]2-(2S, 3S)-(-)-epicatechin (compound 2), were structurally characterized from leaves of Anogeissus pendula. The structures were determined by ultraviolet spectroscopy (UV), proton nuclear magnetic resonance (1H NMR), 13C NMR, and heteronuclear multiple bond correlation. Molinspiration and Osiris property explorer applications were used to predict bioactivity and drug score. Drug scores of 0.08 and 0.05 were predicted for compounds 1 and 2, respectively. Predicted bioactivity scores were high. Due to their molecular weight, chemical structure, and conformation, the newly discovered proanthocyanidins possess an inclination to interact with proteins. Based on this premise, both compounds were subjected to in vitro testing against ruminal enzymes. They exhibited significant inhibition activities (p < 0.01) with a range of half maximal effective concentration (EC50) of 14.80-17.88 mg/mL of glutamic oxaloacetic transaminase in both protozoa and bacteria fractions. The ruminal glutamic pyruvic transaminase activity was significantly inhibited (p < 0.01) from EC50 12.59-16.29 mg/mL, and R-cellulase inhibition was recorded with EC50 18.20-21.98 mg/mL by compounds 1 and 2, respectively. Protease activity decreased with increasing incubation time and concentration of both compounds. The novel proanthocyanidins have potential roles in improving feed conversion ratios and in drug development.

Effect of Vanillin on the Anaesthesia of Crucian Carp: Effects on Physiological and Biochemical Indices, Pathology, and Volatile Aroma Components.

The anaesthetic effect of vanillin on crucian carp was investigated using different concentrations of vanillin, with a nonvanillin control. The effective concentration range of vanillin anaesthesia was determined from the behavioural characteristics of crucian carp during the anaesthesia onset and recovery phases. Physiological and biochemical indices, and the electronic nose response to the fish muscle, were measured over the range of effectiveanaestheticc concentrations. An increased concentration of vanillin shortened the time taken to achieve deep anaesthesia but increased the recovery time. The levels of white blood cells, red blood cells, haemoglobinn, platelets, alanine aminotransferase, alkaline phosphatase, lactate dehydrogenase, phosphorus, potassium, magnesium, total protein, and serum albumin were lower than the control in the vanillin treatment group. Triglycerides and total cholesterol were not significantly affected. Histology showed no effect of vanillin on the liver, except at 1.00 g/L vanillin. Vanillin resulted in a nondose-responsive effect on the gill tissue, increasing the width and spacing of the gill lamellae. E-Nose analysis of the carp-muscle flavour volatiles was able to distinguish between different vanillin treatment concentrations. GC-IMS identified 40 flavour compounds, including 8 aldehydes, 11 alcohols, 10 ketones, 2 esters, and 1 furan. Vanillin had aanaestheticic effect on crucian carp and these findings provide a theoretical basis for improving the transport and experimental manipulation of crucian carp.

Efficient Transfection via an Unexpected Mechanism by Near Neutral Polypiperazines with Tailored Response to Endosomal pH.

Cationic pH-responsive polymers promise to overcome critical challenges in cellular delivery. Ideally, the polymers become selectively charged along the endosomal pathway disturbing only the local membrane and avoiding unintended interactions or cytotoxic side effects at physiological conditions. Polypiperazines represent a novel, hydrophilic class of pH-responsive polymers whose response can be tuned within the relevant pH range (5-7.4). The authors discovered that the polypiperazines are effectively binding plasmid DNA (pDNA) and demonstrate high efficiency in transfection. By design of experiments (DoE), a wide parameter space (pDNA and polymer concentration) is screened to identify the range of effective concentrations for transfection. An isopropyl modified polypiperazine is highly efficient over a wide range of concentrations outperforming linear polyethylenimine (l-PEI, 25kDa) in regions of low N*/P ratios. A quantitative polymerase chain reaction (qPCR) surprisingly revealed that the pDNA within the piperazine-based polyplexes can be amplified in contrast to polyplexes based on l-PEI. The pDNA must therefore be more accessible and bound differently than for other known transfection polymers. Considering the various opportunities to further optimize their structure, polypiperazines represent a promising platform for designing effective soluble polymeric vectors, which are charge-neutral at physiological conditions.

Effects of Gibberellic Acid (GA3) Application on The Plant Growth and Seed Production of Pinto Peanut (Arachis pintoi Krap &amp; Greg)

Pinto peanut (Arachis pintoi Krap &amp; Greg.) is a legume that is usually used as a cover crop, bio mulch in fruit and vegetable plantations, ornamental plants, and animal feed. Pinto peanut has many benefits; through the symbiosis with rhizobacteria they can fix nitrogen, as ground cover can reduce the risk of landslides, inhibit weed growths, and is a source of nectar for bees. Arahcnis pintoi can be propagated vegetatively or generatively, but generative propagation is hard to conduct in the tropics because it takes a long time for the plants to produce seeds. Our study was conducted to determine the effect of GA3 application on seed production of A. pintoi. The experiment was arranged in a single factor randomized complete block design with GA3 concentration of 0, 75, 150, 225, and 300 ppm. An orthogonal polynomial test was conducted to determine the effective concentrations for GA3 on seed formation and seed viability. Harvested seeds were stored for 1 month, then sown on sand medium; the seeds were soaked in 1% KNO3 solution to break seed dormancy prior. Our study showed that the GA3 effective concentration range from 130.69 ppm to 137.16 ppm, indicated by the increase the number of flowers at 105, 120, 135, and 150 DAT up to 28.59% compared to control. GA3 at 126.80 ppm can also increase the number of harvested pods by 18.16%. The effect of GA3 on vegetative growth was shown by the increase in the growth of new individual plants concentration, i.e. 53.25 new individual plant with GA3 application of 141.88 ppm.

Improved PCR by the Use of Disruptors, a New Class of Oligonucleotide Reagents.

As a powerful tool, polymerase chain reaction (PCR) has been indispensable and widely used in a large array of applications. In practice, many factors may affect the overall performance of a PCR. One such factor is the stability of intramolecular secondary structure formed within single-stranded template. The higher the stability of such a structure, the more likely it will have adverse effects on PCR performance. Traditionally, chemical reagents believed to reduce the stability of nucleic acid secondary structures, such as DMSO and betaine, have been used to mitigate their adverse effects on PCR performance. However, these reagents have apparent downsides including increasing replication error rate, inhibiting polymerase activity, and being ineffective against secondary structures of very high stabilities. Disruptors, a new class of oligonucleotide reagents, do not exhibit such downsides. They are specifically designed to target intramolecular secondary structures only without any effect on the replication of other regions of the template. Their effective concentration range for improving PCR performance is well tolerated by PCR. And they are very effective in improving PCR performance on templates that are notoriously difficult to amplify by PCR even in the presence of DMSO or betaine, e.g., the inverted terminal repeat of adeno-associated virus (AAV-ITR). In this chapter, the application of disruptors in PCR is described with AAV-ITR as the example template.