Novel metal enhanced dual-mode fluorometric and SERS aptasensor incorporating a heterostructure nanoassembly for ultrasensitive T-2 toxin detection.

Fluorescent gold (Au) nanostructures have emerged as burgeoning materials to fabricate nanomaterial assemblies which play a vital role in improving the detection sensitivity and specificity for various biomolecules. In this work, a fluorescence labelled (Rhodamine-B-Isothiocyanate) silica shell with Au metal core (AuNPs@PVP@RITC@SiO2) and a graphene-Au nanostar nanocomposite (rGO-AuNS) are presented as a metal enhanced fluorescence (MEF) material and Raman signal enhancer, respectively. Their composite (AuNPs@PVP@RITC@SiO2NPs/rGO-AuNS) was employed as a dual-mode fluorescence (FL) and surface-enhanced Raman scattering (SERS) nanoprobe for selective and sensitive detection of T-2 toxin. To comprehend the dual-modality, a core-shell nanostructure, AuNPs@PVP@RITC@SiO2, was functionalized with an aptamer (donor) and adsorbed on the surface of rGO-AuNS through electrostatic forces and π-π stacking which act as a FL quencher and SERS signal enhancer. When exposed to T-2 toxin, the apt-AuNPs@PVP@RITC@SiO2NPs move away from the surface of rGO-AuNS, resulting in the restoration of FL and reduction of the SERS signal. There was distinct linearity between the T-2 toxin concentration and the dual FL and SERS signals with lower limits of detection (LOD) of 85 pM and 12 pM, as compared to the previous methods, respectively. The developed FL and SERS aptasensor presented excellent recovery ratio and RSD in wheat and maize, respectively, as compared with the standard ELISA method. The complementary performances of the developed stratagem revealed a high correlation between the FL and SERS sensing modes with exquisite detection properties.

Analyzing the determinants of sustainability of China Pakistan Economic Corridor (CPEC) projects: an interpretive structural modelling (ISM) approach.

China Pakistan Economic Corridor (CPEC) is a game changer initiative of South Asian Pacific Rim. It has great importance for almost all Asian countries. Its success is expected to dictate the economic development of the stakeholders. The aim of this study is to evaluate the essential determinants deriving the sustainability of CPEC projects. The design of the study comprises of the review of literature, data collection, and analysis. Population under study is the folk of stakeholders of CPEC. Sampling envisages on purposive sampling design, i.e., 14 experts from within the stakeholders. Primary data is collected in the field setting through a survey questionnaire appropriate for the study. ISM is used for modelling and MICMAC for analysis and classification using inductive approach. The findings of the literature survey show that there are 23 prime determinants of sustainability of CPEC projects. The results of ISM show that 13 determinants are at Level-I, nine at Level-II, and one determinant namely "economic globalization" is at Level-III being the most critical and driving determinant. The findings of MICMAC show that only one determinant is classified in independent quadrant, and all the remaining determinants are in linkage quadrant, whereas, no determinant is shown in autonomous and/or dependence quadrant. But most of the determinants have potential to be classified in dependent and independent quadrants. It is intimately evident that the results of MICMAC corroborate the results of ISM. It is useful for folk of the stakeholders by way of developing an understanding about the multitude of determinants, intra-determinant relations, prioritizing the determinants for policy decisions, and/or for building future studies. This study has some limitations, e.g., the study uses qualitative approach and answers what and how questions that do not quantify the relations or tell the cause of indicated relations.

Effectiveness Of Local Anaesthetic In Reducing Postoperative Pain At Port Site After Laparoscopic Cholecystectomy.

Background: Laparoscopy has become the highest quality level way to deal with cholecystectomy since its inception 30 years preceding, and is perhaps the most normally performed general surgeries. Pain being a significant issue after laparoscopic cholecystectomy bringing about extended admissions or readmissions. With significant varieties in pain relieving conventions an integrated approach is important to diminish pain. The aim of this study is to assess the effectiveness of Bupivacaine as local anesthetic agent at port sites after laparoscopic cholecystectomy. Method: Study population of 84 patients was divided into control group (receiving no local anesthetic) and study group (receiving Bupivacaine as local anesthetic). Visual analogue scale was used to quantify and compare pain perceived by each group; at fixed intervals of 6, 12 and 24 hours after shifting of the patients back to the ward. Results: Each group comprised 42 patients. At 6 hours post operative pain score in study group, 4.5±0.32 was significantly lower than in control group, 7.6±0.41 (p<0.05). Though pain assessments at 12 and 24 hours didn't reveal any significant differences among the two groups; postoperative requirement of Tramadol was significantly (p<0.05) lower in study group (92±0.064mg) in comparison to control group (158±0.21mg). Conclusion: Use of long-acting local anesthetic injections at port sites after laparoscopic cholecystectomy significantly lowers pain during first 6 hours post operatively and also lowers narcotic analgesics requirements during post operative period.

Synthesis and Investigation of the Analgesic Potential of Enantiomerically Pure Schiff Bases: A Mechanistic Approach.

Schiff bases are a class of organic compounds with azomethine moiety, exhibiting a wide range of biological potentials. In this research, six chiral Schiff bases, three 'S' series (H1−H3) and three 'R' series (H4−H6), were synthesized. The reaction was neat, which means without a solvent, and occurred at room temperature with a high product yield. The synthesized compounds were evaluated for analgesic potential in vivo at doses of 12.5 and 25 mg/kg using acetic-acid-induced writhing assay, formalin test, tail immersion and hot plate models, followed by investigating the possible involvement of opioid receptors. The compounds H2 and H3 significantly (*** p < 0.001) reduced the writhing frequency, and H3 and H5 significantly (*** p < 0.001) reduced pain in both phases of the formalin test. The compounds H2 and H5 significantly (*** p < 0.001) increased latency at 90 min in tail immersion, while H2 significantly (*** p < 0.001) increased latency at 90 min in the hot plate test. The 'S' series Schiff bases, H1−H3, were found more potent than the 'R' series compounds, H4−H6. The possible involvement of opioid receptors was also surveyed utilizing naloxone in tail immersion and hot plate models, investigating the involvement of opioid receptors. The synthesized compounds could be used as alternative analgesic agents subjected to further evaluation in other animal models to confirm the observed biological potential.

Preserving Brain LPC-DHA by Plasma Supplementation Attenuates Brain Injury after Cardiac Arrest.

OBJECTIVE: Cardiac arrest (CA) is a major health burden with brain damage being a significant contributor to mortality. We found lysophosphatidylcholine (LPC), including a species containing docosahexaenoic acid (LPC-DHA), was significantly decreased in plasma post-CA, supplementation of which significantly improved neurological outcomes. The aim of this study is to understand the protective role of LPC-DHA supplementation on the brain post-CA. METHODS: We first evaluated associations between the plasma level of LPC-DHA and neurological injury and outcomes of human patients with CA. We then utilized a rat CA model and cell cultures to investigate therapeutic and mechanistic aspects of plasma LPC-DHA supplementation. RESULTS: We found that decreased plasma LPC-DHA was strongly associated with neurological outcomes and disappearance of the difference between gray and white matter in the brain after CA in human patients. In rats, the decreased plasma LPC-DHA was associated with decreased levels of brain LPC-DHA after CA, and supplementing plasma LPC-DHA normalized brain levels of LPC-DHA and alleviated neuronal cell death, activation of astrocytes, and expression of various inflammatory and mitochondrial dynamics genes. We also observed deceased severity of metabolic alterations with LPC-DHA supplementation using untargeted metabolomics analysis. Furthermore, LPC treatment showed a similar protective effect for neurons and astrocytes in mixed primary brain cell cultures. INTERPRETATION: The observed neuroprotection accompanied with normalized brain LPC-DHA level by plasma supplementation implicate the importance of preventing the decrease of brain LPC-DHA post-CA for attenuating brain injury. Furthermore, the data supports the causative role of decreased plasma LPC-DHA for brain damage after CA. ANN NEUROL 2022;91:389-403.

Green Synthesis and Characterization of Carboxymethyl Cellulose Fabricated Silver-Based Nanocomposite for Various Therapeutic Applications.

PURPOSE: The current study proposed the simple, eco-friendly and cost-effective synthesis of carboxymethyl cellulose (CMC) structured silver-based nanocomposite (CMC-AgNPs) using Syzygium aromaticum buds extract. METHODS: The CMC-AgNPs were characterized by ultraviolet (UV) spectroscopy, scanning electron microscopy (SEM), transmission electron microscopy (TEM), X-ray diffraction (XRD), Fourier transmission infra-red (FTIR), energy-dispersive X-ray (EDX), and dynamic light scattering (DLS) techniques. The synthesized nanocomposites were evaluated for their bactericidal kinetics, in-vivo anti-inflammatory, anti-leishmaniasis, antioxidant and cytotoxic activities using different in-vitro and in-vivo models. RESULTS: The spherical shape nanocomposite of CMC-AgNPs was synthesized with the mean size range of 20-30 nm, and the average pore diameter is 18.2 nm while the mean zeta potential of -31.6 ± 3.64 mV. The highly significant (P < 0.005) antibacterial activity was found against six bacterial strains with the ZIs of 24.6 to 27.9 mm. More drop counts were observed in Gram-negative strains after 10 min exposure with CMC-AgNPs. Significant damage in bacterial cell membrane was also observed in atomic force microscopy (AFM) after treated with CMC-AgNPs. Nanocomposite showed highly significant anti-inflammatory activity in cotton pellet induced granuloma model (Phase I) in rats with the mean inhibitions of 43.13% and 48.68% at the doses of 0.025 and 0.05 mg/kg, respectively, when compared to control. Reduction in rat paw edema (Phase II) was also highly significant (0.025 mg/kg; 42.39%; 0.05 mg/kg, 47.82%). At dose of 0.05 mg/kg, CMC-AgNPs caused highly significant decrease in leukocyte counts (922 ± 83), levels of CRP (8.4 ± 0.73 mg/mL), IL-1 (177.4 ± 21.3 pg/mL), IL-2 (83.7 ± 11.5 pg/mL), IL-6 (83.7 ± 11.5 pg/mL) and TNF-α (18.3 ± 5.3 pg/mL) as compared to control group. CMC-AgNPs produced highly effective anti-leishmaniasis activity with the viable Leishmania major counts decreased up to 36.7% within 24 h, and the IC50 was found to be 28.41 µg/mL. The potent DPPH radical scavenging potential was also observed for CMC-AgNPs with the IC50 value of 112 µg/mL. Furthermore, the cytotoxicity was assessed using HeLa cell lines with the LC50 of 108.2 µg/mL. CONCLUSION: The current findings demonstrate positive attributes of CMC fabricated AgNPs as a promising antibacterial, anti-inflammatory, anti-leishmaniasis, and antioxidant agent with low cytotoxic potential.

Characterization of the Mitochondrial Genome of a Wheat AL-Type Male Sterility Line and the Candidate CMS Gene.

Heterosis utilization is very important in hybrid seed production. An AL-type cytoplasmic male sterile (CMS) line has been used in wheat-hybrid seed production, but its sterility mechanism has not been explored. In the present study, we sequenced and verified the candidate CMS gene in the AL-type sterile line (AL18A) and its maintainer line (AL18B). In the late uni-nucleate stage, the tapetum cells of AL18A showed delayed programmed cell death (PCD) and termination of microspore at the bi-nucleate stage. As compared to AL18B, the AL18A line produced 100% aborted pollens. The mitochondrial genomes of AL18A and AL18B were sequenced using the next generation sequencing such as Hiseq and PacBio. It was found that the mitochondrial genome of AL18A had 99% similarity with that of Triticum timopheevii, AL18B was identical to that of Triticum aestivum cv. Chinese Yumai. Based on transmembrane structure prediction, 12 orfs were selected as candidate CMS genes, including a previously suggested orf256. Only the lines harboring orf279 showed sterility in the transgenic Arabidopsis system, indicating that orf279 is the CMS gene in the AL-type wheat CMS lines. These results provide a theoretical basis and data support to further analyze the mechanism of AL-type cytoplasmic male sterility in wheat.

Synergistic anti-oxidative effects of Pongamia pinnata against nickel mediated by Rhizobium pisi and Ochrobacterium pseudogrignonense.

Nickel is widely spread by different anthropogenic activities and shows toxicity for plant growth and development. Whether rhizobia symbiotically fix nitrogen can eliminate or reduce nickel toxic effect on plant or not is still unknown. This study was aimed to investigate the effect of different rhizobia genus inoculation on growth, nitrogen fixing ability, metal accumulation and enzymatic antioxidative balance of Pongamia pinnnaa. Inoculation with Rhizobium pisi and Ochrobacterium pseudogrignonense increased the all the growth parameters both in 0 and 40 mg/kg nickel as comparison with control. Only shoot length increased in presence of nitrogen as compared with no supply of nitrogen. Nitrogen content also increased both in rhizobia inoculation as compared to no nitrogen supply and non-inoculation control, respectively. Nickel uptake was higher in shoots and leaves but lower in roots in case of inoculation as compared to non-inoculation control. Rhizobia inoculation improved the plant antioxidant capacity by increasing the activity of enzymatic scavengers catalase (CAT), superoxide dismutase (SOD), peroxidase (POD) and ascorbate (GR). However, 40 mg/kg of nickel adding showed mostly effect on the activity CAT, SOD, POD in leaves. All the enzymatic activity showed a significant increase in absence of nitrogen supply as compared nitrogen supply. Our results suggested that rhizobia inoculation effectively mediated nickel stress for legume plants by increasing nitrogen supplement and inducing antioxidant capacity.

Evaluation of rapid polymyxin Nordmann Poirel test for detection of polymyxin resistance in clinical isolates of Enterobacteriaceae.

Polymyxins play a significant role against carbapenem-resistant Enterobacteriaceae (CRE). A total of 121 clinical samples yielded growth of CRE that were included in the study. Rapid Polymyxin NP test was performed on all the isolates as described by Nordmann P et al. and results were compared with broth microdilution method. Majority of the isolates were Klebsiella pneumoniae (70.2%) followed by Escherichia coli (17.4%). A total of 71 isolates were found resistant and 50 as susceptible by broth microdilution. Sensitivity and specificity of rapid polymyxin NP test were found to be 97.2% and 100%, respectively. Our study concluded that rapid polymyxin NP test is reliable and can be used as an alternative to broth microdilution in resource limited settings.

Antibacterial, anticoagulant and cytotoxic evaluation of biocompatible nanocomposite of chitosan loaded green synthesized bioinspired silver nanoparticles.

Present work reports the green synthesis of chitosan functionalized silver nanoparticles (CS-AgNPs) using ethanolic buds extract of Sygyzium aromaticum. CS-AgNPs were characterized physically, evaluated for antibacterial, anticoagulant and antiplatelet activities, and toxicity profile. The physical characterization of CS-AgNPs was done by UV/vis, SEM, TEM, FTIR and EDX. The sphericity was found uniform. FTIR and EXD showed noninterfering few impurities. The antibacterial activity against VRSA (ZI, 23.2 ± 0.51 mm) and MRSA (ZI, 25.8 ± 0.32 mm) were determined. The rise in bleeding and thromboplastin was observed highly significant while increased in prothrombin and activated partial prothrombin time in significant manner at both the doses of CS-AgNPs (0.025 mg/kg and 0.05 mg/kg). Reduction in the levels of fibrinogen was also highly significant. Platelet aggregation decreased at high dose of CS-AgNPs i.e. 55.14 ± 8.25% (arachidonic acid) and 13.06 ± 2.17% (collagen). Thrombin antithrombin (TAT) complex activity was found highest for CS-AgNPs. Cytotoxicity was assessed using HeLa cell lines (LC50; 125 µg/ml) and brine shrimp lethality tests (LC50; 518 µg/ml). The work suggests that green synthesized chitosan functionalized silver nanoparticles may be utilized as an effective antibacterial agent and anticoagulant with low toxicity. The current findings will open a new window for nanomedicine development and future clinical application.

Selected Micronutrients: An Option to Boost Immunity against COVID-19 and Prevent Adverse Pregnancy Outcomes in Pregnant Women: A Narrative Review.

The coronavirus disease-19 (COVID-19) negatively affects immune system. It is linked with adverse pregnancy outcomes. These complications may be linked with the infections mediated deficiency of micronutrients in pregnant women. COVID-19 cause's malabsorption of micronutrients thereby increases the risk of their deficiency. Both micronutrients deficiencies and poor micronutrients intake can compromise immune function and may increase the risk of pregnancy complications associated with COVID-19 infection. Vita-min A, C, D, E, and selected minerals iron (Fe), selenium (Se), and zinc (Zn) are the micronutrients essential for immuno-competency and play a significant role in the prevention of adverse pregnancy outcomes. Immune function and pregnancy outcomes can be improved by adequate intake of micronutrients in diet or in supplements form. Based on regulatory links between viral infection, micronutrients, immunity, and pregnancy outcomes, this review highlights the role of micronutrients in boosting immunity to reduce or prevent pregnancy complications in COVID-19 infected women.

Antimicrobial potentials of medicinal plant's extract and their derived silver nanoparticles: A focus on honey bee pathogen.

Infectious (or Communicable) diseases are not only the past but also the present problem in developing as well as developed countries. It is caused by various pathogenic microbes like fungi, bacteria, parasites and virus etc. The medicinal plants and nano-silver have been used against the pathogenic microbes. Herbal medicines are generally used for healthcare because they have low price and wealthy source of antimicrobial properties. Like medicinal plants, silver nanoparticles also have emergent applications in biomedical fields due to their immanent therapeutic performance. Here, we also explore the various plant parts such as bark, stem, leaf, fruit and seed against Gram negative and Gram-positive bacteria, using different solvents for extraction i.e. methanol, ethyl acetate, chloroform, acetone, n. hexane, butanol, petroleum ether and benzene. Since ancient to date most of the countries have been used herbal medicines, but in Asia, some medicinal plants are commonly used in rural and backward areas as a treatment for infectious diseases. In this review, we provide simple information about medicinal plants and Silver nanoparticles with their potentialities such as antiviral, bactericidal and fungicidal. Additionally, the present review to highlights the versatile applications of medicinal plants against honey bee pathogen such as fungi (Ascosphaera apis), mites (Varroa spp. and Tropilaelaps sp.), bacteria (Melissococcus plutonius Paenibacillus larvae), and microsporidia (Nosema apis and Nosema ceranae). In conclusion, promising nonchemical (plant extracts) are innocuous to adult bees. So, we strongly believed that this effort was made to evaluate the status of medicinal plants researches globally.

A novel bioassay based on aptamer-functionalized magnetic nanoparticle for the detection of zearalenone using time resolved-fluorescence NaYF4: Ce/Tb nanoparticles as signal probe.

Zearalenone (ZEN) is a non-steroidal estrogenic mycotoxin produced by fungi on stored grains. The earlier detection methods used for ZEN rely on expensive equipment, time-consuming sample preparation and temperature sensitive antibodies. The current work, proposed a novel strategy based on ZEN aptamer labeled with amine-functionalized magnetic nanoparticle (MNPs) as a capture probe and time-resolved fluorescence (TRFL) nanoparticles labeled with complementary DNA (cDNA) as a signal probe. Under the optimized conditions, TRFL intensity at 544 nm was used to measure ZEN (R2 = 0.9920) in the range of 0.001-10 ng mL-1 and limits of detection (LOD) for proposed method was 0.21 pg mL-1. The specificity of bioassay was also determined by using other mycotoxins (OTA, AFB2, DON and Patulin) and results showed that the aptamer are specific to recognize only ZEN. The analytical applications of the present bioassay in maize and wheat samples were also examined and results were compared with existing methods. Based on these findings, it is suggested to use current rapid and simple bioassay for the determination of ZEN in food and agricultural products.

In vitro antioxidant and hepatoprotective activities of Paeonia emodi (Wall.) rhizome methanol extract and its phenolic compounds rich fractions.

The present study aimed to quantify the total phenolic content in Paeonia emodi rhizome methanol extract and its fractions and then evaluate the in vitro antioxidant and hepatoprotective activities of fractions rich in phenolic compounds. Maximum quantity of total phenolic content was observed in butanol (112.08±5.5 mg GAE/g dw) and chloroform fraction (107.0±3.5 mg GAE/g dw) followed by methanol extract (94.2±4.4 mg GAE/g dw), aqueous fraction (92.9±2.5 mg GAE/g dw), ethyl acetate (62.3±8.3 mg GAE/g dw) and n-hexane fraction (51.6±7.2 mg GAE/g dw). The fractions rich in total phenolic content were evaluated for in vitro antioxidant activity based on 2, 2-diphenyl-1-picryl-hydrazyl (DPPH) scavenging assay. The butanol and chloroform fraction showed significantly (P<0.05) higher radical scavenging activity with IC50 values of 6.5 and 7.05±2.5 ppm respectively. Positive correlation (R square=0.95) was observed between total phenolic content and in vitro antioxidant activity. The fractions rich in phenolic compounds were also evaluated for their hepatoprotective activity in paracetamol intoxicated mice. Five days oral administration of these fractions at a dose of 300 mg/kg body weight restored the serum ALT, AST and ALP levels of paracetamol intoxicated mice to normal level. From the results of the present research it was concluded that the butanol and chloroform fractions of P. emodi rhizome methanol extract are rich in phenolic compounds and strong antioxidant and effective in attenuation of hepatotoxicity.

Use of in vivo biotinylation for chromatin immunoprecipitation.

This unit describes a system for expression of biotinylated proteins in mammalian cells in vivo, and its application to chromatin immunoprecipitation (ChIP). The system is based on co-expression of the target protein fused to a short biotin acceptor domain, together with the biotinylating enzyme BirA from Escherichia coli. The superior strength of the biotin-avidin interaction in the modified ChIP protocol presented here allows one to employ more stringent washing conditions, resulting in a better signal/noise ratio. Methods for interpreting the data obtained from ChIP samples analyzed by qPCR, and methods for testing the efficiency of biotinylation using a streptavidin gel-shift are also presented. In addition, a complementary method, based on isothermal multiple strand displacement amplification (IMDA) of circular concatemers generated from the DNA fragments obtained after ChIP, is described. This method helps to decrease bias in DNA amplification and is useful for the analysis of complex mixtures of DNA fragments typically generated in miniscale ChIP experiments.