Investigations of Limeum Indicum Plant for Diabetes Mellitus and Alzheimer's Disease Dual Therapy: Phytochemical, GC-MS Chemical Profiling, Enzyme Inhibition, Molecular Docking and In-Vivo Studies.

Limeum indicum has been widely utilized in traditional medicine but no experimental work has been done on this herb. The primary objective of this study was to conduct a phytochemical analysis and assess the multifunctional capabilities of aforementioned plant in dual therapy for Alzheimer's disease (AD) and Type 2 diabetes (T2D). The phytochemical screening of ethanol, methanol extract, and their derived fractions of Limeum indicum was conducted using GC-MS, HPLC, UV-analysis and FTIR. The antioxidant capacity was evaluated by DPPH method. The inhibitory potential of the extracts/fractions against α-, ß-glucosidase acetylcholinesterase (AChE), butyrylcholinesterase (BChE) and monoaminine oxidases (MAO-A & B) was evaluated. Results revealed that acetonitrile fraction has highest inhibitory potential against α-glucosidase (IC50=68.47±0.05 µg/mL), methanol extract against ß-glucosidase (IC50=91.12±0.07 µg/mL), ethyl acetate fraction against AChE (IC50=59.0±0.02 µg/mL), ethanol extract against BChE (28.41±0.01 µg/mL), n-hexane fraction against MAO-A (IC50=150.5±0.31 µg/mL) and methanol extract for MAO-B (IC50=75.95±0.13 µg/mL). The docking analysis of extracts\fractions suggested the best binding scores within the active pocket of the respective enzymes. During the in-vivo investigation, ethanol extract produced hypoglycemic effect (134.52±2.79 and 119.38±1.40 mg/dl) after 21 days treatment at dose level of 250 and 500 mg/Kg. Histopathological findings further supported the in-vivo studies.

Public Health and Health System's Responsiveness During the 2022 Floods in Pakistan: What Needs to Be Done?

OBJECTIVE: In 2022, Pakistan witnessed unprecedented flooding, submerging one-third of the country under-water, ruining millions of houses, taking lives, afflicted injuries, and displacing scores of people. Our study documents not only the public health problems that have arisen due to this natural calamity but also the state of health systems' response. METHODS: We conducted a qualitative study asking key questions around prevalent health problems, health-care seeking, government's response, resource mobilization, and roadmap for the future. We purposively selected 16 key frontline health workers for in-depth interviews. RESULTS: Waterborne and infectious diseases were rampant posing huge public health challenges. Disaster mitigation efforts and relief operations were delayed and not at scale to cover the entire affected population. Moreover, a weak economy, poverty, and insufficient livelihoods compounded the tribulations of floods. Issues of leadership and governance at state level resulted in disorganized efforts and response. CONCLUSIONS: Pakistan is famous for its philanthropy; however, lack of transparency and accountability, the actual benefits seldom reach the beneficiaries. Such climatic disasters necessitate a more holistic approach and a greater responsiveness of the health system. In addition to health services, the state must respond to financial, social, and infrastructural needs of the people suffering from the calamity.

Potential of psychrotolerant rhizobacteria for the growth promotion of wheat (Triticum aestivum L.).

Wheat is the second most important staple crop grown and consumed worldwide. Temperature fluctuations especially the cold stress during the winter season reduces wheat growth and grain yield. Psychrotolerant plant growth-promoting rhizobacteria (PGPR) may improve plant stress-tolerance in addition to serve as biofertilizer. The present study aimed to isolate and identify PGPR, with the potential to tolerate cold stress for subsequent use in supporting wheat growth under cold stress. Ten psychrotolerant bacteria were isolated from the wheat rhizosphere at 4 °C and tested for their ability to grow at wide range of temperature ranging from -8 °C to 36 °C and multiple plant beneficial traits. All bacteria were able to grow at 4 °C to 32 °C temperature range and solubilized phosphorus except WR23 at 4 °C, whereas all the bacteria solubilized phosphorus at 28 °C. Seven bacteria produced indole-3-acetic acid at 4 °C, whereas all produced indole-3-acetic acid at 28 °C. Seven bacteria showed the ability to fix nitrogen at 4 °C, while all the bacteria fixed nitrogen at 28 °C. Only one bacterium showed the potential to produce cellulase at 4 °C, whereas four bacteria showed the potential to produce cellulase at 28 °C. Seven bacteria produced pectinase at 4 °C, while one bacterium produced pectinase at 28 °C. Only one bacterium solubilized the zinc at 4 °C, whereas six bacteria solubilized the zinc at 28 °C using ZnO as the primary zinc source. Five bacteria solubilized the zinc at 4 °C, while seven bacteria solubilized the zinc at 28 °C using ZnCO3 as the primary zinc source. All the bacteria produced biofilm at 4 °C and 28 °C. In general, we noticed behavior of higher production of plant growth-promoting substances at 28 °C, except pectinase assay. Overall, in vitro testing confirms that microbes perform their inherent properties efficiently at optimum temperatures rather than the low temperatures due to high metabolic rate. Five potential rhizobacteria were selected based on the in vitro testing and evaluated for plant growth-promoting potential on wheat under controlled conditions. WR22 and WR24 significantly improved wheat growth, specifically increasing plant dry weight by 42% and 58%, respectively. 16S rRNA sequence analysis of WR22 showed 99.78% similarity with Cupriavidus campinensis and WR24 showed 99.9% similarity with Enterobacter ludwigii. This is the first report highlighting the association of C. campinensis and E. ludwigii with wheat rhizosphere. These bacteria can serve as potential candidates for biofertilizer to mitigate the chilling effect and improve wheat production after field-testing.

Flavonoids from Pistacia chinensis subsp. integerrima with leishmanicidal activity: computational and experimental evidence.

Pistacia chinensis subsp. integerrima is a valuable medicinal plant as its parts and extracts found application for treating diarrhea, fever, liver disorders, asthma, and inflammation. In this study, we report the leishmanicidal activity of sakuranetin, spinacetin, and patuletin extracted from P. chinensis. The tested compounds revealed a strong anti-leishmanial activity in vitro against Leishmania major showing IC50 values of 7.98 ± 0.16 µM, 9.23 ± 0.23 µM 11.09 ± 0.87 µM for sakuranetin, spinacetin, and patuletin, respectively. Moreover, to explore the potential mechanism(s) by which the compounds may act, computational docking studies were performed against dihydrofolate reductase and pteridine reductase, showing that the flavonoids could target these two key enzymes to exploit their leishmanicidal activity. In accordance with in vitro results, patuletin was highlighted as the most promising compound of the set, and binding energy values of -6.72 and -6.74 kcal/mol were computed for the two proteins, respectively.

A detailed biochemical characterization, toxicological assessment and molecular docking studies of Launaea fragilis: An important medicinal xero-halophyte.

Launaea fragilis (Asso) Pau (Family: Asteraceae) is a wild medicinal plant that has been used in the folklore as a potential treatment for numerous ailments such as skin diseases, diarrhea, infected wounds, inflammation, child fever and hepatic pain. This study explored the chemical constitution, in-vivo toxicity, antimicrobial, antioxidant, and enzyme inhibition potential of ethanolic extract of L. fragilis (EELF). Additionally, in-silico docking studies of predominant compounds were performed against in-vitro tested enzymes. Similarly, in-silico ADMET properties of the compounds were performed to determine their pharmacokinetics, physicochemical properties, and toxicity profiles. The EELF was found rich in TFC (73.45 ± 0.25 mg QE/g) and TPC (109.02 ± 0.23 mg GAE/g). GC-MS profiling of EELF indicated the presence of a total of 47 compounds mainly fatty acids and essential oil. EELF showed no toxicity or growth retardation in chicks up to 300 mg/kg with no effect on the biochemistry and hematology of the chicks. EELF gave promising antioxidant activity through the CUPRAC method with an IC50 value of 13.14 ± 0.18 µg/ml. The highest inhibition activity against tyrosinase followed by acetylcholinesterase and α-Glucosidase was detected. Similarly, the antimicrobial study revealed the extract with good antibacterial and antiviral activity. A good docking score was observed in the in silico computational study of the predominant compounds. The findings revealed L. fragilis as a biocompatible, potent therapeutic alternative and suggest isolation and further in vivo pharmacological studies.

Antioxidant and Hepatoprotective Activities of Acacia jacquemontii Stem Extract against High-fat and CCl4-induced Liver Injury in Rat's Model.

BACKGROUND: Chronic liver injury leads to liver inflammation and fibrosis, activating myofibroblasts in the liver and secreting extracellular matrix proteins that make the fibrous scar. OBJECTIVES: The purpose of our study was to characterize the polyphenolic content present in Acacia jacquemontii stem and evaluate its antioxidant and hepatoprotective activity. METHODS: The phenolic contents in Acacia jacquemontii polyphenolic extract (AJPPE) were characterized using high-performance liquid chromatography (HPLC). The hepatoprotective and antioxidant activity of AJPPE were determined through biochemical parameters (ALT, AST, and ALP), lipid profile (TC, TG, HDL, and LDL), antioxidant biomarkers (SOD, LPO, GSH, and CAT), anti-fibrotic activity (collagen deposition), and histopathological analysis. RESULTS: HPLC analysis of AJPPE showed the presence of polyphenols, including chlorogenic acid, P-coumaric acid, caffeic acid, and kaempferol, in a remarkable therapeutic range. Results of the in vivo analysis showed a significant decrease in the level of lipid profile, including LDL (low-density lipoprotein), TC (total cholesterol), triglycerides, liver function markers (AST, ALT, and ALP), collagen deposition and significantly increased the level of anti-oxidative biomarkers (CAT, SOD, LPO, and GSH) by using AJPPE. CONCLUSION: The above-mentioned results have shown that AJPPE possesses significant antioxidative and hepatoprotective effects. Furthermore, histopathological results also supported the antioxidant and hepatoprotective potential of AJPPE.

Controllable synthesis and stabilization of Tamarix aphylla-mediated copper oxide nanoparticles for the management of Fusarium wilt on musk melon.

Excessive use of pesticides and mineral fertilizers poses a serious threat to ecoenvironment sustainability and human health. Nano pesticides or Nano fungicides have attained great attention in the field of agriculture due to their unique characteristics, by improving crop growth with enhancing pathogenesis-related defense system. However, there is a need to develop a sustainable mechanism for the synthesis of fungicides which replace the chemical pesticides to avoid their hazardous impact. Here in, Tamarix aphylla mediated CuO-Nanoparticles (NPs) were synthesized, characterized and their activity was evaluated under in-vitro and in-vivo conditions. The structural and elemental analysis of NPs were carried out by using X-ray powder diffraction (XRD), Fourier transformed infrared spectroscopy (FTIR), UV-visible spectrophotometer, Scanning electron microscope (SEM) and Transmission electron microscope (TEM). In the greenhouse, at an optimum concentration of 50 mg/L reduced disease severity very effectively and enhanced plant growth. Application of NPs also assisted in the induction of systemic response of defense-related genes in melon. Under In vitro condition at 100 mg/L significantly reduced mycelial growth (84.5%) by directly acting on the pathogenic cell wall. Our work confirmed that dosedependent concentration of T. aphylla extract based biological CuO-NPs enhance plant growth and help to effectively resist against F. oxysporum infection. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-022-03189-0.

Vascular Protective Effect and Its Possible Mechanism of Action on Selected Active Phytocompounds: A Review.

Vascular endothelial dysfunction is characterized by an imbalance of vasodilation and vasoconstriction, deficiency of nitric oxide (NO) bioavailability and elevated reactive oxygen species (ROS), and proinflammatory factors. This dysfunction is a key to the early pathological development of major cardiovascular diseases including hypertension, atherosclerosis, and diabetes. Therefore, modulation of the vascular endothelium is considered an important therapeutic strategy to maintain the health of the cardiovascular system. Epidemiological studies have shown that regular consumption of medicinal plants, fruits, and vegetables promotes vascular health, lowering the risk of cardiovascular diseases. This is mainly attributed to the phytochemical compounds contained in these resources. Various databases, including Google Scholar, MEDLINE, PubMed, and the Directory of Open Access Journals, were searched to identify studies demonstrating the vascular protective effects of phytochemical compounds. The literature had revealed abundant data on phytochemical compounds protecting and improving the vascular system. Of the numerous compounds reported, curcumin, resveratrol, cyanidin-3-glucoside, berberine, epigallocatechin-3-gallate, and quercetin are discussed in this review to provide recent information on their vascular protective mechanisms in vivo and in vitro. Phytochemical compounds are promising therapeutic agents for vascular dysfunction due to their antioxidative mechanisms. However, future human studies will be necessary to confirm the clinical effects of these vascular protective mechanisms.

Gastroprotective Effect of Polypeptide-K Isolated from Momordica charantia's Seeds on Multiple Experimental Gastric Ulcer Models in Rats.

Peptic ulcer disease is a multifactorial disorder and is the most significant public health concern nowadays. Previous study showed that essential oil extracted from Momordica charantia's seed exhibited gastroprotective effect. However, the evidence for the gastroprotective effect of its active compound, polypeptide K (PPK), remains unclear. This study aimed to examine the preventive effect of PPK against different experimental gastric lesions models in rats. The possible gastroprotective effect of PPK was assessed in hydrochloride ethanol- and indomethacin-induced gastric ulcer models in Sprague Dawley rats and was further evaluated macroscopically and microscopically. Pyloric ligation experiments were used to investigate gastric secretion. Oral administration of PPK at all concentrations (10, 25, and 50 mg/kg) showed significant (p < 0.05) reduction in total area of lesion in both hydrochloride ethanol- and indomethacin-induced gastric ulcer models. The highest inhibition rate was seen in PPK dose of 50 mg/kg with 64.9% and 72.2% on hydrochloride ethanol and indomethacin models, respectively. Microscopically, PPK preserved the normal architectures of the gastric tissues from being damaged by hydrochloride ethanol and indomethacin. Further, in the pyloric ligation studies, PPK significantly (p < 0.05) decreased the ulcer area where the highest protection was exhibited by 50 mg/kg with 70% inhibition rate. Moreover, all concentrations of PPK also significantly (p < 0.05) enhanced the gastric wall mucus secretion. Collectively, this study demonstrated the gastroprotective effect of PPK on hydrochloride ethanol- and indomethacin-induced gastric ulcer models. The possible mechanism might be associated with enhanced mucus secretion and thus lowering the total acidity.

Phytochemical profiling, in vitro and in vivo anti-inflammatory, analgesic and antipyretic potential of Sesuvium sesuvioides (Fenzl) Verdc. (Aizoaceae).

Sesuvium sesuvioides (Fenzl) Verdc is traditionally used in the treatment of inflammatory diseases such as arthritis and gout The aim of present study was to assess the possible anti-inflammatory, analgesic and antipyretic potential of the methanol extract of Sesuvium sesuvioides (SsCr) to prove scientifically its folklore use in the inflammatory diseases and to screen its total antioxidant capacity by multiple methods and phytocompounds by GC-MS. The preliminary phytochemical studies showed the presence of phenols, flavonoids, glycosides, coumarin, terpenoids, saponins, fats and carbohydrates in crude extract. The total phenolic contents (27.31 ± 0.28 mg GAE/g) and total flavonoids (3.58 ± 0.12 mgRE/g) values were observed. The antioxidant capacity of SsCr showed significant DPPH, ABTS, CUPRAC, FRAP, PBD and metal chelating results. GC-MS analysis displayed the phytoconstituents with anti-inflammatory potentials such as 2-methoxy-4-vinylphenol, vanillin, umbelliferone, methyl ferulate, palmitoleic acid, methyl palmitate and phytol. SsCr presented noteworthy HRBC membrane stability with maximum inhibition of cell hemolysis (47.79%). In carrageenan-induced hind paw edema assay result showed dose-dependent anti-inflammatory action. SsCr presented significant (p < 0.05) analgesic activity in hot-plate and tail flicking tests similarly it also showed the noteworthy inhibition in pain latency against formalin induced analgesia at 1st and 2nd phases. SsCr reduced the acetic acid-induced writhes at different doses (250, 500 and 750 mg). Results of antipyretic activity of SsCr extract were significant at 500 and 750 mg. The results of in vitro and in vivo experimental studies verified the anti-inflammatory, analgesic and antipyretic potential of Sesuvium sesuvioides and supported the folklore uses of this plant.

Essential Oils Derived from Momordica charantia Seeds Exhibited Antiulcer Activity against Hydrogen Chloride/Ethanol and Indomethacin.

Momordica charantia (MC) is popular for its medicinal uses especially for treating diabetic-related complications. However, the antiulcer activity of essential oil derived from the seeds has not been systematically studied. This study aims to evaluate the gastroprotective activities of essential oil derived from the seed of MC induced by hydrochloride acid/ethanol (HCl/EtOH) and indomethacin and pylorus-ligation model. Gastric ulceration was induced by oral administration of HCl/EtOH solution or indomethacin on day 7 after animals have been pretreated with testing compounds. The first group received just distilled water and the second group received ranitidine (100 mg/kg). Groups 3, 4, and 5 received 10, 50, and 100 mg/kg of essential oil based on their body weight (10 mL/kg), respectively. Macroscopically, pretreatment of essential oil extracted from MC significantly decreased ulceration induced by HCl/EtOH and indomethacin in vivo. Microscopically, essential oil also significantly suppressed the formation of edema, epithelial disruption, and mucosa erosions. Moreover, essential oil significantly elevated the pH without decreasing the total acidity of the gastric juice and was able to increase the amount of adherent mucus compared to control. Current results provide scientific basis to the ethno-pharmacological usage of the MC in preventing ulcer formation induced by HCl/EtOH and indomethacin.

Continuous medication monitoring: A clinical model to predict adherence to medications among chronic kidney disease patients.

BACKGROUND: An adherence model is required to optimise medication management among chronic kidney disease (CKD) patients, as current assessment methods overestimate the true adherence of CKD patients with complex regimens. An approach to assess adherence to individual medications is required to assist pharmacists in addressing non-adherence. OBJECTIVE: To develop an adherence prediction model for CKD patients. METHODS: This multi-centre, cross-sectional study was conducted in 10 tertiary hospitals in Malaysia using simple random sampling of CKD patients with ≥1 medication (sample size = 1012). A questionnaire-based collection of patient characteristics, adherence (defined as ≥80% consumption of each medication for the past one month), and knowledge of each medication (dose, frequency, indication, and administration) was performed. Continuous data were converted to categorical data, based on the median values, and then stratified and analysed. An adherence prediction model was developed through multiple logistic regression in the development group (n = 677) and validated on the remaining one-third of the sample (n = 335). Beta-coefficient values were then used to determine adherence scores (ranging from 0 to 7) based on the predictors identified, with lower scores indicating poorer medication adherence. RESULTS: Most of the 1012 patients had poor medication adherence (n = 715, 70.6%) and half had good medication knowledge (n = 506, 50%). Multiple logistic regression analysis determined 4 significant predictors of adherence: ≤7 medications (constructed score = 2, p < 0.001), ≤3 co-morbidities (constructed score = 1, p = 0.015), absence of complementary/alternative medicine use (constructed score = 1, p = 0.003), and knowledge score ≥80% (constructed score = 3, p < 0.001). A higher total constructed score from the prediction model indicated a higher likelihood of adherence (odds ratio [OR]: 2.41; 95% confidence interval [CI]: 2.112-2.744; p < 0.001). The area under the receiver operating characteristic (ROC) curve of the developed model (n = 677) had good accuracy (ROC: 0.867, 95% CI: 0.840-0.896; p < 0.001). The validated model (n = 335) also had good accuracy (ROC: 0.812, 95% CI: 0.765-0.859; p < 0.001). There was no significant difference between the development and validation groups (p = 0.11, Z-value:1.62, standard error: 0.034). CONCLUSION: The score constructed from the medication adherence prediction model for CKD patients had good accuracy and could be useful for identifying patients with a higher risk of non-adherence, to ensure optimised adherence management.

Polyphenolics and triterpenes presence in chloroform extract of Dicranopteris linearis leaves attenuated paracetamol-induced liver intoxication in rat.

INTRODUCTION: Water-soluble, but not lipid-soluble, extract of Dicranopteris linearis leaves has been proven to possess hepatoprotective activity. The present study aimed to validate the hepatoprotective and antioxidant activities, and phytoconstituents of lipid-soluble (chloroform) extract of D. linearis leaves. METHODS: The extract of D. linearis leaves (CEDL; 50, 250 and 500 mg/kg) was orally administered to rats for 7 consecutive days followed by the oral administration of 3 g/kg PCM to induce liver injury. Blood was collected for liver function analysis while the liver was obtained for histopathological examination and endogenous antioxidant activity determination. The extract was also subjected to antioxidant evaluation and phytochemicals determination via phytochemical screening, HPLC and UPLC-HRMS analyses. RESULTS: CEDL exerted significant (p < 0.05) hepatoprotective activity at 250 and 500 mg/kg and significantly (p < 0.05) reversed the PCM-induced decrease in rat's liver endogenous antioxidant (catalase and superoxide dismutase) level. CEDL possessed a high antioxidant capacity when measured using the ORAC assay, but a low total phenolic content value and radical scavenging activity as confirmed via several radical scavenging assays, which might be attributed particularly to the presence of triterpenes. Phytochemicals screening demonstrated the presence of triterpenes and flavonoids, while UPLC-HRMS analysis showed the presence of polyphenols belonging to the hydroxybenzoic acids, hydroxycinammates and flavonoid groups. DISCUSSION AND CONCLUSION: Lipid-soluble bioactive compounds of CEDL demonstrated hepatoprotective effect against PCM intoxication partly via the modulation of the endogenous antioxidant defense system, and exerted high antioxidant capacity. Further investigation is warranted to identify the potential hepatoprotective leads from CEDL for future drug development.

Effect of increasing pre-pubertal concentrate levels on reproductive and lactation performance of Nili-Ravi Buffalo heifers.

The objective of this study was to determine the effect of increasing total nutrient supply by increasing level of concentrate supplementation in forage-based diet of growing Nili-Ravi buffalo replacement heifers on pre-pubertal growth rate and its impact on age at puberty and post-pubertal productive and reproductive performance. Twenty-eight Nili-Ravi buffalo heifers of 7 ± 1 month age were divided into 3 groups. The treatments were 3 levels of concentrate (crude protein (CP) = 17.9%, metabolisable energy (ME) = 2.66 Mcal/kg), i.e. 0.5, 1, and 1.5% of the body weight referred as low, medium, and high treatments, respectively. Irrespective of pre-pubertal treatment, all the heifers received similar feed after puberty up to completion of their first lactation. The results of the study showed a similar response across the treatments in age, weight, and condition at puberty, weight at calving, lactation length, and yield. The medium and high treatments resulted in poor reproductive performance because of higher services per conception and less number of heifers confirmed conceived compared with low treatment. The low treatment resulted in less mortality compared with medium and high treatments. The feeding cost per animal at puberty, conception, and calving was lowest for treatment low as compared with medium and high. In conclusion, the results of the study showed that Nili-Ravi buffalo heifers achieved puberty at 23 months with feeding of concentrate (CP = 17.9%, ME = 2.66 Mcal/kg) during pre-pubertal stage at the rate of 0.5% of the body weight with better reproductive performance.

Microbiota in cancer development and treatment.

PURPOSE: Human microbiota comprises of a variety of organisms ranging from bacterial species to viruses, fungi, and protozoa which are present on the epidermal and mucosal barriers of the body. It plays a key role in health and survival of the host by regulation of the systemic functions. Its apparent functions in modulation of the host immune system, inducing carcinogenesis and regulation of the response to the cancer therapy through a variety of mechanisms such as bacterial dysbiosis, production of genotoxins, pathobionts, and disruption of the host metabolism are increasingly becoming evident. METHODS: Different electronic databases such as PubMed, Google Scholar, and Web of Science were searched for relevant literature which has been reviewed in this article. RESULTS: Characterization of the microbiome particularly gut microbiota, understanding of the host-microbiota interactions, and its potential for therapeutic exploitation are necessary for the development of novel anticancer therapeutic strategies with better efficacy and lowered off-target side effects. CONCLUSION: In this review, the role of microbiota is explained in carcinogenesis, mechanisms of microbiota-mediated carcinogenesis, and role of gut microbiota in modulation of cancer therapy.

Effects of Different Pollens on Primary Metabolism and Lignin Biosynthesis in Pear.

To investigate the effect of pollination on the fruit quality of 'Dangshan Su' pear, 'Dangshan Su' was fertilized by the pollen of 'Wonhwang' (Pyrus pyrifolia Nakai.) (DW) and 'Jingbaili' (Pyrus ussuriensis Maxim.) (DJ). The analysis of primary metabolites was achieved through untargeted metabolomics, and the quantitative analysis of intermediate metabolites of lignin synthesis was undertaken using targeted metabolomics. The untargeted metabolomics analysis was performed via gas chromatography-mass spectrometry (GC-MS). The targeted metabolomics analysis was performed using ultra-high-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) under the multiple reaction monitoring (MRM) mode. The results showed that the metabolite content was significantly different between DW and DJ. Compared with that in DJ, the sugar and amino acid content in DW was higher and the fatty acid content was lower at 47 days after pollination (DAPs), and the sugar, amino acid, and fatty acid content in DW was lower at 63 DAPs. The intermediate metabolites of lignin synthesis were analyzed using the orthogonal partial least squares discriminant analysis (OPLS-DA) model, and the differential metabolites at 47 DAPs were p-coumaric acid, ferulic acid, sinapaldehyde, coniferyl alcohol, and sinapyl alcohol. The differential significant metabolite at 63 DAPs was p-coumaric acid. At 47 DAPs and 63 DAPs, the p-coumaric acid level was significantly different, and the p-coumaric acid content was positively correlated with lignin synthesis. The pollination pollen affects the quality of 'Dangshan Su' pear fruit through regulation of the sugar, amino acid, and fatty acid content; at the same time, regulating the levels of intermediate metabolites of lignin synthesis, especially the p-coumaric acid content, to affect lignin synthesis ultimately affects the stone cell content and improves the quality of the pears.

Albizia chevalier based Ag nanoparticles: Anti-proliferation, bactericidal and pollutants degradation performance.

The eco-friendly biosynthesis of silver nanoparticles (AgNps) from bark extract of Albizia chevalier are reported here for their anti-proliferative, antibacterial and pollutant degradation potentials. The synthesized AgNps were characterized by FTIR spectroscopy, transmission electron microscopy (TEM), field emission scanning electron microscopy (FESEM), energy dispersive X-rays spectrometry (EDS) and X-ray diffraction studies. The TEM and FESEM images show a monodispersed spherical shaped particles of approximately 30 nm. Crystalline peaks were obtained for the synthesized AgNps in XRD spectrum. The AgNps were investigated for in vitro anticancer and antibacterial activities and its potential to degrade 4-nitrophenol (4-NP) and congo red dye (CR). The MTT results shows a significant dose-dependent antiproliferation effect of the AgNps on the cell lines HepG2, MDA-MB-231 and MFC7. The effect was found more pronounced in MDA-MB-231 as compared to MFC-7 cell lines. The antibacterial results indicated 99 and 95% killing of Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) respectively, after 24 h of incubation with the AgNps. The AgNps were found to speed up the reductive degradation of 4-NP and CR dye, which give an alternative route for the removal of toxic organic pollutants from the wastewater. The synthesized AgNps were not only used as a bactericidal and anticancer agent, but also effectively used for the reductive degradation of carcinogenic compounds which are listed as the priority pollutants. Therefore, AgNps have the potential for the treatment of various cancers, bacterial infections and for industrial detoxification of wastewater.

In vitro investigation of cytotoxic and antioxidative activities of Ardisia crispa against breast cancer cell lines, MCF-7 and MDA-MB-231.

BACKGROUND: Ardisia crispa Thunb. D.C is used mostly in some parts of the Asian region by traditional practitioners to treat certain diseases associated with oxidative stress and inflammation including cancer and rheumatism. In Malaysia, it is popularly known as 'Mata Ayam' and local traditional practitioners believed that the root of the plant is therapeutically beneficial. METHODS: The cytotoxic effect of hydromethanolic extract of A. crispa and its solvents partitions (ethyl acetate and aqueous extracts) against breast cancer cells were evaluated by using MTT assay. The cells were treated with concentration of extracts ranging from 15.63 µg/mL- 1000 µg/mL for 72 h. The quantification of phenolic and flavonoid contents of the extracts were carried out to determine the relationship between of phytochemical compounds responsible for cytotoxic and antioxidative activities. The antioxidant capacity was measured by DPPH and ABTS free radical scavenging assay and expressed as milligram (mg) Trolox equivalent antioxidant capacity per 1 g (g) of tested extract. RESULTS: The hydromethanolic and ethyl acetate extracts showed moderate cytotoxic effect against MCF-7 with IC50 values of 57.35 ± 19.33 µg/mL, and 54.98 ± 14.10 µg/mL, respectively but aqueous extract was inactive against MCF-7. For MDA-MB-231, hydromethanolic, ethyl acetate and aqueous extracts exhibited weak cytotoxic effects against MDA-MB-231 with IC50 values more than 100 µg/mL. The plant revealed high total phenolic content, total flavonoid and antioxidant capacity. CONCLUSION: The response of different type of breast cancer cell lines towards A. crispa extract and its partitions varied. Accordingly, hydromethanolic and ethyl acetate extracts appear to be more cytotoxic to oestrogen receptor (ER) positive breast cancer than oestrogen receptor (ER) negative breast cancer. However, aqueous extract appears to have poor activity to both types of breast cancer. Besides that, hydromethanolic and ethyl acetate extracts exhibit higher TPC, TFC and antioxidant capacity compared to aqueous extract. Synergistic effect of anticancer and antioxidant bioactives compounds of A. crispa plausibly contributed to the cytotoxic effects of the extract.

iTRAQ-Based Identification of Proteins Related to Lignin Synthesis in the Pear Pollinated with Pollen from Different Varieties.

Most pears in Anhui Province are a kind of self-incompatible fruit whose quality is strongly influenced by the male pollen. The proteomic variation of Dangshan Su pollinated by different varieties was analysed using the isobaric tag for relative and absolute quantitation (iTRAQ) to investigate the effect of pollination by different varieties on the pear lignin pathway. Among the 3980 proteins identified from the two samples, 139 proteins were identified as differentially expressed proteins (DEPs). Of these proteins, laccase-4 (LAC4), was found to be related with lignin synthesis, and ß-glucosidase 15 (BGLU15) and peroxidase 47 (PER47) were involved in the phenylpropanoid pathway. Moreover, the lignin and stone cell contents were lower in DW (Dangshan Su pollinated by Wonhwang) than those in DJ (Dangshan Su pollinated by Jingbaili). The effect of pollination on the synthesis of lignin through the regulation of the expression of PER47, BGLU15 and LAC4 ultimately affects the formation of stone cells and the fruit quality. We report for the first time that different pollinations influence the protein expression profile in the Dangshan Su pear, and this result provides some new epididymal targets for regulating the synthesis of lignin, regulating the content of stone cells and improving the quality of the pears.

Synergistic gastroprotective activity of methanolic extract of a mixture of Melastoma malabathricum and Muntingia calabura leaves in rats.

BACKGROUND: Melastoma malabathricum L. (family Melastomaceae; MM) and Muntingia calabura L. (family Elaeocarpaceae; MC) have been separately reported to possess gastroprotective activity. In an attempt to develop a pharmaceutical product with antiulcer potential, the synergistic gastroprotective activity of methanolic extract of a mixture of MM and MC (MMMC) at various ratios was evaluated in rat models. METHODS: Rats were pre-treated orally with 2% Tween 80 (vehicle), 100 mg/kg ranitidine (reference drug) or MMMC (ratios of 1:1, 1:3 and 3:1 (v/v); doses of 15, 150 or 300 mg/kg) and then subjected to the ethanol-induced gastric ulcer or pyloric ligation assays. Stomach of rats from the former assay was collected and subjected to the macroscopic and microscopic observations, and enzymatic and non-enzymatic antioxidant studies while the gastric juice content and tissue from the latter assay were subjected to the antisecretory activity study. The UHPLC analysis of MMMC was also performed. RESULT: MMMC, in the ratio 1:1, demonstrated the most effective (P < 0.001) gastroprotective activity indicated by the highest reduction in ethanol-induced ulcer area formation. These macroscopic findings were supported by the microscopic observations. Except for pH and total acidity, MMMC also significantly (P < 0.001) reduced the volume of gastric content but increased the gastric wall mucus content in the pyloric-ligation test. MMMC also demonstrated remarkable antioxidant activity indicated by the highest total phenolic content (TPC) value and oxygen radical absorbance capacity (ORAC) activity with the recorded IC50 value of approximately 53 µg/mL for the 2,2- diphenyl-1-picrylhydrazyl (DPPH) scavenging activity. MMMC also improved the catalase (CAT), superoxide dismutase (SOD), glutathione (GSH), prostaglandin E2 (PGE2) and malondialdehyde (MDA) activities of the gastric tissue intoxicated by ethanol. UHPLC analysis of MMMC confirmed the presence several flavonoid-based bioactive compounds. CONCLUSION: MMMC, at the ratio of 1:1 (v/v), exerts gastroprotective activity partly by activating its antisecretory and antioxidant activities, and via modulation of the gastric tissue endogenous antioxidant system.