Development and validation of a GC-MS method for analysis of Dithiocarbamate Fungicide residues in the Spices Cardamom (Elettaria cardamomom) and black pepper (Piper nigrum).

Dithiocarbamates constitute an important class of broad-spectrum antifungal compounds used extensively in agriculture, including in the cultivation of spices. Maximum residue limits for these compounds have been enforced by several importing countries in international food trade. Validation of analytical methods for dithiocarbamates in spices have not been reported previously. A quick and sensitive method for estimation of total dithiocarbamates as carbon disulphide (CS2) using GC-MS in two major spices, viz. small cardamom (Elettaria cardamomom) and black pepper (Piper nigrum) was optimized and validated. Dithiocarbamate residues in these spice matrices were extracted and subjected to acid hydrolysis followed by reduction to CS2, which was then quantitatively absorbed into isooctane and analysed using GC-MS, employing selected ion monitoring and post-run mid-column backflush technique. For fortification levels from 0.1 to 1.0 mg kg- 1, recoveries obtained ranged from 75 to 98% in cardamom and 76-98% in black pepper, with intra-day precision (RSDr) < 12% and inter-day precision (RSDR) < 15% in all cases. Limit of Quantification of 0.05 mg kg- 1 was achieved in both spices. It was found that there was negligible interference in quantitative accuracy due to essential oils present in the two spices studied. Matrix effect was seen to be suppressive in the two spices studied, and sufficiently low to exclude the use of matrix-matched calibration in routine quantitative analysis. The optimized analytical method was found to be suitable for evaluation of compliance of real samples against the Codex maximum residue limits for cardamom and black pepper. Safety evaluation for human consumption, based on the incidence of Dithiocarbamate residues, was performed in real samples of cardamom and black pepper. This method offers the possibility of extending applicability to other spices also. Supplementary information: The online version contains supplementary material available at 10.1007/s13197-022-05462-9.

Comparative Oral Absorption of Curcumin in a Natural Turmeric Matrix with Two Other Curcumin Formulations: An Open-label Parallel-arm Study.

Curcuminoids are the major bioactive molecules in turmeric, and poor bioavailability deters them from being the major components of many health and wellness applications. This study was conducted to assess the bioavailability of a completely natural turmeric matrix formulation (CNTMF) and compare its bioavailability with two other commercially available formulations, namely, curcumin with volatile oil (volatile oil formulation) and curcumin with phospholipids and cellulose (phospholipid formulation) in healthy human adult male subjects (15 each group) under fasting conditions. Each formulation was administrated orally as a single 500-mg dose in capsule form, and blood samples were analyzed by liquid chromatography mass spectrometry at various time intervals up to 24 h. The ingestion of the CNTMF was very well absorbed and resulted in a mean curcuminoids plasma Cmax of 170.14 ng/mL (Tmax  = 4 h) compared with 47.54 ng/mL and 69.63 ng/mL for the volatile oil (Tmax  = 3 h) and phospholipid (Tmax  = 2.25 h) formulations, respectively. The extent of absorption of total curcuminoids in the blood for the CNTMF was 6× greater than volatile oil formulation and 5× greater than phospholipids formulation. The results of this study indicate that curcumin in a natural turmeric matrix exhibited greater bioavailability than the two comparator products. Copyright © 2017 John Wiley & Sons, Ltd.

Aryne-Induced Novel Tandem 1,2-Addition/(3+2) Cycloaddition to Generate Imidazolidines and Pyrrolidines.

A new "single-flask" method was developed for the synthesis of imidazolidines and pyrrolidines with high stereoselectivity. First, a Schiff base was arylated with an aryne. Second, an intramolecular proton transfer took place from the methylene position to the anionic aryne ring. Third, the resultant ylide reacted with a second equivalent of the same Schiff base in situ or an electron-deficient alkene through a (3+2) cycloaddition. These sequential tandem 1,2-addition/(3+2) cycloaddition reactions led to the desired heterocycles in 63-88% yields.

Possibly pathogenic bacteria in aerosols and foams as a result of aeration remediation in a polluted urban waterway.

Newtown Creek is a tributary of the Hudson River Estuary. It has a legacy of both industrial pollution and sewage pollution and has been designated a Superfund site. To ameliorate the chronically low levels of dissolved oxygen detected in the Creek, the New York City Department of Environmental Protection has been installing aerators. The abundance of various bacteria in the aerosols, foams, and water, at two sites in the Creek, was studied before, during, and after the aeration process. Additionally, aerosols and dispersed foams created by the aeration process were sampled and cultured to determine what unique taxa of bacteria could be grown and identified. Taxa including Actinobacteria and Firmicutes were prevalent in cultures taken from aerosols, whereas Gammaproteobacteria were prevalent in cultures taken from foam. Campylobacteria was found to have a significant presence in both samples taken after the aerators were turned off. These taxa include potentially pathogenic bacteria and are therefore of particular concern.

Fiber-Reinforced-Phospholipid Vehicle-Based Delivery of l-Ascorbic Acid: Development, Characterization, ADMET Profiling, and Efficacy by a Randomized, Single-Dose, Crossover Oral Bioavailability Study.

l-ascorbic acid (AA) or vitamin C is a crucial nutrient needed for optimal health. However, being unable to be synthesized by the body, it is thus necessary to be included in health care products. Moreover, AA is one of the antioxidants that occur naturally, which is used in pharmaceutical and food products as an antioxidant additive. However, AA is vulnerable to environmental settings and undergoes oxidative degradation to dehydroascorbic acid and further to inactive products. Therefore, new research strategies and approaches are required to augment its stability. The objective of this study is to develop and characterize a fiber-reinforced-phospholipid (FRP) matrix-based vehicle, Zeal-AA, for the delivery of AA and optimize the oral bioavailability of the obtained AA powder using an efficacy study by open-label, randomized, single-dose, two-treatment, two-sequence, two-period, two-way crossover. The structural and surface morphologies were analyzed by Fourier transform infrared spectroscopy, transmission electron microscopy, scanning electron microscopy, and differential scanning calorimetry studies. Encapsulation efficiency, mean particle size, size distribution, ζ-potential measurements, and ADMET profiling revealed the potential delivery system for AA. AUC0-t was found to be 55.23 (mg/dL) for Zeal-AA, whereas it was 9.38 (mg/dL) for AA, and C max was found to be 6.69 (mg/dL) for Zeal-AA, whereas it was 1.23 (mg/dL) for AA, with a fold difference of bioavailability in terms of AUC found to be 5.9 fold. The results show that a single oral dose of Zeal-AA is capable of rising the AA levels in the body relative to the control up to 24 h.

Preparation and Characterization of Liposomal ß-Caryophyllene (Rephyll) by Nanofiber Weaving Technology and Its Effects on Delayed Onset Muscle Soreness (DOMS) in Humans: A Randomized, Double-Blinded, Crossover-Designed, and Placebo-Controlled Study.

Delayed onset muscle soreness (DOMS) is a complex spreading out, which is related to swelling of muscles, tenderness, rigidity, pain, disruption of muscle fiber, alteration in the kinematics of joint, acute tissue damage, and reduction in power and strength. ß-Caryophyllene (BCP), a potent phytocannabinoid, could play an important role in managing DOMS because of its wide diversity of biological activities, particularly its anti-inflammatory activity; however, its poor stability in light, temperature, high volatility, and insolubility can restrict the medical practices. In this study, liposomal ß-caryophyllene (Rephyll) was designed and established in powder form constructed by the nanofiber weaving technology to improve the bioavailability of BCP with improved stability. Rephyll was characterized by Fourier transform infrared spectroscopy, transmission electron microscopy, and differential scanning calorimetry studies. Encapsulation efficiency, loading capacity, and in vitro release studies revealed that Rephyll can be an auspicious drug delivery arrangement for BCP. The effects of Rephyll were evaluated by a randomized, double-blinded, crossover-designed, placebo-controlled study. The oral consumption of Rephyll significantly reduced the pain visual assessment score, revealing that Rephyll effectively reduced DOMS with improved recovery without any side effects due to the bioavailable form of the phytocannabinoid BCP in the liposomal powder formulation.

Chitosan and polyvinyl alcohol nanocomposites with cellulose nanofibers from ginger rhizomes and its antimicrobial activities.

The agro-industrial waste obtained after the isolation of bio-constituents from ginger is available in abundance. In the present study, the effective isolation of ginger nanofibers (GNF) was carried out by acid hydrolysis and high pressure homogenization to get cellulose nanofibers with 100 to 200 nm width. Bionanocomposites were also prepared by reinforcing different ratios of (1% to 7%) GNF with chitosan (CS) and polyvinyl alcohol (PVA) matrices by solvent cast method and the 5% GNF with CS and PVA resulted a high mechanical strength composites than others. The surface morphology and structural analysis of the composites were identified by Scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), Differential scanning calorimetry (DSC) and X-ray diffraction (XRD) methods. The inhibitory effect of 5% GNF bionanocomposites against Bacillus cereus, Escherichia coli, Staphylococcus aureus and Salmonella typhimurium indicated good antibacterial activity of the nanocomposites due to the addition of GNF in the biopolymer matrices. The use of GNF will help to increase the economic values of agricultural waste and the characteristic properties of GNF derived bionanocomposites could be possibly used in medical and packaging areas.

Non-Curcuminoids from Turmeric and Their Potential in Cancer Therapy and Anticancer Drug Delivery Formulations.

Over the past decades curcuminoids have been extensively studied for their biological activities such as antiulcer, antifibrotic, antiviral, antibacterial, antiprotozoal, antimutagenic, antifertility, antidiabetic, anticoagulant, antivenom, antioxidant, antihypotensive, antihypocholesteremic, and anticancer activities. With the perception of limited toxicity and cost, these compounds forms an integral part of cancer research and is well established as a potential anticancer agent. However, only few studies have focused on the other bioactive molecules of turmeric, known as non-curcuminoids, which are also equally potent as curcuminoids. This review aims to explore the comprehensive potency including the identification, physicochemical properties, and anticancer mechanism inclusive of molecular docking studies of non-curcuminoids such as turmerones, elemene, furanodiene (FN), bisacurone, germacrone, calebin A (CA), curdione, and cyclocurcumin. An insight into the clinical studies of these curcumin-free compounds are also discussed which provides ample evidence that favors the therapeutic potential of these compounds. Like curcuminoids, limited solubility and bioavailability are the most fragile domain, which circumscribe further applications of these compounds. Thus, this review credits the encapsulation of non-curcuminoid components in diverse drug delivery systems such as co-crystals, solid lipid nanoparticles, liposomes, microspheres, polar-non-polar sandwich (PNS) technology, which help abolish their shortcomings and flaunt their ostentatious benefits as anticancer activities.

A novel bioavailable hydrogenated curcuminoids formulation (CuroWhite™) improves symptoms and diagnostic indicators in rheumatoid arthritis patients - A randomized, double blind and placebo controlled study.

Rheumatoid arthritis (RA) is an inflammatory disease that cause chronic pain, disability and joint destruction. The present placebo controlled randomized study aimed to evaluate the efficacy of a novel hydrogenated curcuminoid formulation-CuroWhite™, in rheumatoid arthritis (RA) patients. Twenty four RA patients were randomized in 1:1:1 ratio to receive 250 mg, 500 mg CuroWhite or placebo as one capsule a day, over a period of three months. Improvement in the ACR response, changes in disease activity assessed using the DAS 28 score, change in physical function assessed on change in ESR, CRP, RF values were evaluated before and after the study. Results suggested that patients who received CuroWhite both low and high doses reported statistically significant changes in their clinical symptoms towards end of the study when compared with placebo. There were significant changes in DAS28 (50-64%) VAS (63-72%) ESR (88-89%), CRP (31-45%) RF (80-84%) values and ACR response for CuroWhite groups in comparison with placebo. Thus, CuroWhite acts as the analgesic and anti-inflammatory product for management of RA by the reduction of the inflammatory action which was confirmed by improvement in ESR, CRP, VAS, RF, DAS-28 and ACR responses. CuroWhite was significantly effective against RA with highly safe without serious side effects and well tolerated.

Differential expression of the TFIIIB subunits Brf1 and Brf2 in cancer cells.

BACKGROUND: RNA polymerase (pol) III transcription is specifically elevated in a variety of cancers and is a target of regulation by a variety of tumor suppressors and oncogenes. Accurate initiation by RNA pol III is dependent on TFIIIB. In higher eukaryotes, two forms of TFIIIB have been characterized. TFIIIB required for proper initiation from gene internal RNA pol III promoters is comprised of TBP, Bdp1, and Brf1. Proper initiation from gene external RNA pol III promoters requires TBP, Bdp1, and Brf2. We hypothesized that deregulation of RNA polymerase III transcription in cancer may be a consequence of altered TFIIIB expression RESULTS: Here, we report: (1) the TFIIIB subunits Brf1 and Brf2 are differentially expressed in a variety of cancer cell lines: (2) the Brf1 and Brf2 promoters differ in activity in cancer cell lines, and (3) VAI transcription is universally elevated, as compared to U6, in breast, prostate and cervical cancer cells. CONCLUSION: Deregulation of TFIIIB-mediated transcription may be an important step in tumor development. We demonstrate that Brf1 and Brf2 mRNA are differentially expressed in a variety of cancer cells and that the Brf2 promoter is more active than the Brf1 promoter in all cell lines tested. We also demonstrate, that Brf1-dependent VAI transcription was significantly higher than the Brf2-dependent U6 snRNA transcription in all cancer cell lines tested. The data presented suggest that Brf2 protein expression levels correlate with U6 promoter activity in the breast, cervical and prostate cell lines tested. Interestingly, the Brf1 protein levels did not vary considerably in HeLa, MCF-7 and DU-145 cells, yet Brf1 mRNA expression varied considerably in breast, prostate and cervical cancer cell lines tested. Thus, Brf1 promoter activity and Brf1 protein expression levels did not correlate well with Brf1-dependent transcription levels. Taken together, we reason that deregulation of Brf1 and Brf2 expression could be a key mechanism responsible for the observed deregulation of RNA pol III transcription in cancer cells.

A Randomized Single Dose Parallel Study on Enhancement of Nitric Oxide in Serum and Saliva with the Use of Natural Sports Supplement in Healthy Adults.

Sports supplements that stimulate the production of nitric oxide (NO) are widely promoted agents in the sports nutrition domain, and nitric oxide plays an important role to enhance the cardiovascular and physical fitness of the sports participants. The purpose of the study is to investigate whether oral intake of a sports nutritional supplement (Fitnox) is able to increase nitrate (NO3-) and nitrite (NO2-) levels in blood serum and saliva of healthy adults. Fitnox is a unique blend of Kaempferia parviflora methoxy flavones, pomegranate peel polyphenols, and Moringa oleifera leaf saponins. Twenty-four healthy male adults were equally divided and underwent the double-blind, placebo-controlled clinical trial with a single oral dose of sports nutrition formulation (250 mg capsules); blood and saliva samples were analyzed at different time intervals by high-performance liquid chromatography (HPLC). After administration of Fitnox (250 mg capsule as single dose), NO3- and NO2- levels in serum and saliva were found to be significantly higher (p <.05) than in the placebo group in 24 hours. Pharmacokinetic parameters such as the area under the plasma concentration-time curve extrapolated to infinity (AUC0-inf), AUC calculated to the last measured concentration (AUC0-t), maximum drug serum concentrations (Cmax), time of maximum concentration in serum observed (Tmax), and time required for the concentration of the drug to reach half of its original value (Thalf) were also statistically significant (p <.05) compared with the placebo. The results indicate that a single oral dose of Fitnox is able to increase the NO3- and NO2- levels considerably in the body relative to placebo for at least 12 hours. Therefore, Fitnox can improve the overall performance of sport participants and enhance physical endurance.

A Toxicological Evaluation of a Standardized Hydrogenated Extract of Curcumin (CuroWhite™).

A series of toxicological investigations were conducted in order to evaluate the genotoxic potential and repeated-dose oral toxicity of CuroWhite, a proprietary extract of curcumin that has been hydrogenated and standardized to not less than 25% hydrogenated curcuminoid content. All tests were conducted in general accordance with internationally accepted standards. The test item was not mutagenic in the bacterial reverse mutation test or in vitro mammalian chromosomal aberration test, and no in vivo genotoxic activity was observed in rat bone marrow in the micronucleus test. A 90-day repeated-dose study was conducted in male and female Sprague-Dawley rats. Two mortalities occurred in the main and satellite high-dose groups and were determined due to gavage error. No organ specific or other toxic effects of the test item were observed up to the maximum dose of 800 mg/kg bw/day, administered by gavage. NOAEL was, therefore, estimated as 800 mg/kg bw/day.

Oral toxicity study of sports nutritional powder in Wistar rats: A 90 day repeated dose study.

Fitnox is a newly developed dietary ingredient for physical endurance composed of the extracts of Moinga oleifera leaf (45-50%), Kaempferia parviflora (black ginger) root (15-20%) and Punica granatum peel (25-30%). The aim of this study was to assess the subchronic oral toxicity of Fitnox (test substance) - in Wistar albino rats. Forty rats equally divided into 4 groups (control male, control female, treatment male and treatment female) administrated the test substance at 1000 mg/kg per rat daily for 90 days. All the animals were observed for body weight, mortality and clinical observations during the entire study. Results revealed no significant changes between the control and Fitnox treated groups. Based on the results, it was concluded that orally administered Fitnox to rats (dose of 1000 mg/kg per rat, orally-90 days) is safe with no drug-related toxicity was observed during the study period. Thus, the no-observed adverse effect level (NOAEL) for the present study is evaluated to be 1000 mg/kg body weight in both the sexes.

A Novel Highly Bioavailable Curcumin Formulation Improves Symptoms and Diagnostic Indicators in Rheumatoid Arthritis Patients: A Randomized, Double-Blind, Placebo-Controlled, Two-Dose, Three-Arm, and Parallel-Group Study.

Rheumatoid arthritis (RA) is an autoimmune, chronic systemic inflammatory disorder. The long-term use of currently available drugs for the treatment of RA has many potential side effects. Natural phytonutrients may serve as alternative strategies for the safe and effective treatment of RA, and curcuminoids have been used in Ayurvedic medicine for the treatment of inflammatory conditions for centuries. In this study, a novel, highly bioavailable form of curcumin in a completely natural turmeric matrix was evaluated for its ability to improve the clinical symptoms of RA. A randomized, double-blind, placebo-controlled, three-arm, parallel-group study was conducted to evaluate the comparative efficacy of two different doses of curcumin with that of a placebo in active RA patients. Twelve patients in each group received placebo, 250 or 500 mg of the curcumin product twice daily for 90 days. The responses of the patients were assessed using the American College of Rheumatology (ACR) response, visual analog scale (VAS), C-reactive protein (CRP), Disease Activity Score 28 (DAS28), erythrocyte sedimentation rate (ESR), and rheumatoid factor (RF) values. RA patients who received the curcumin product at both low and high doses reported statistically significant changes in their clinical symptoms at the end of the study. These observations were confirmed by significant changes in ESR, CPR, and RF values in patients receiving the study product compared to baseline and placebo. The results indicate that this novel curcumin in a turmeric matrix acts as an analgesic and anti-inflammatory agent for the management of RA at a dose as low as 250 mg twice daily as evidenced by significant improvement in the ESR, CRP, VAS, RF, DAS28, and ACR responses compared to placebo. Both doses of the study product were well tolerated and without side effects.

Preparation of a novel bioavailable curcuminoid formulation (Cureit™) using Polar-Nonpolar-Sandwich (PNS) technology and its characterization and applications.

Health benefits of curcuminoid are highly limited due to their poor aqueous solubility, very low systemic bioavailability, fast metabolic alterations and rapid elimination. In this study, a novel bioavailable curcuminoid formulation Cureit™ was prepared by using Polar-Nonpolar-Sandwich (PNS) technology with complete natural turmeric matrix (CNTM). The synthesized bioavailable curcuminoid formulation Cureit™ was characterizations by Nuclear magnetic resonance spectroscopy (NMR), scanning electron microscopy (SEM), X-ray diffraction (XRD), Fourier transform infra-red (IR), current-voltage (I-V) study, Quadrupole Time-of-Flight Mass Spectrometry (Q-TOF), differential scanning calorimeter (DSC) and thermogravimetric analysis (TGA). NMR study showed the presence of hydrogen bonding interactions with curcuminoids, polar and non-polar compounds in the PNS technology. SEM images indicated that Cureit™ was almost spherical and well dispersed with rough morphology, and separated with three layers of PNS formulation. The chemical profile of Cureit™ was analyzed by Q-TOF confirmed the presence of curcuminoids (curcumin, demethoxycurcumin and bismethoxycurcumin), lactones, sesquiterpenes and their derivatives derived from polar layer, aromatic turmerone, dihydroturmerone, turmeronol, curdione and bisacurone derived from non-polar layer. IR, XRD, DSC and TGA also confirmed the presence of curcuminoids with high stability in the PNS formulation. Various biological activities of Cureit™ were also discussed.

Acute and subchronic oral toxicity studies of hydrogenated curcuminoid formulation 'CuroWhite' in rats.

Hydrogenated curcuminoids are the major metabolites of the curcumin and 'CuroWhite' is a unique blend of hydrogenated curcuminoids encapsulated with ß-cyclodextrin. There is no particular scientific evidence for the toxicology regarding the hydrogenated curcuminoids, so the present work reports the results of the studies investigating the acute (single dose) and subchronic (repeatedly 90 days) oral toxicity of the CuroWhite in Sprague Dawley rats. For acute oral toxicity testing a sighting study was conducted on female rats in a sequential manner to allow selection of the appropriate starting dose for the main study. In acute toxicity, the dosage was 2000 mg/kg body weight for four female rats. In the sub-chronic study, rats of both sexes divided into three groups and each group were orally treated with CuroWhite daily at 200, 400 and 800 mg/kg for 90 days consecutively. No evidence of treatment related toxicity was detected during the study. Thus, data analysis of mortality, body weight gain, feed consumption, clinical observations, hematology, organ weights and histopathological findings did not show significant differences between control and treated groups. It is concluded that CuroWhite orally administered to rats was safe and no drug-related toxicity was detected even at the highest doses investigated in both acute (2000 mg/kg) and subchronic toxicity (200, 400 and 800 mg/kg) studies. Based on the study, the no-observed-adverse-effect level (NOAEL) value could be considered as 800 mg/kg per day in both the sexes. These results indicate that CuroWhite can be generally regarded as safe for use as a food additive.

Direct and Indirect Costs of Pediatric Gastroenteritis in Vellore, India.

OBJECTIVE: To determine costs of pediatric gastroenteritis in out-patient and in-patient facilities. METHODS: Cross-sectional survey of children with acute gastroenteritis attending out-patient clinic (n=30) or admitted in the ward (n=30) for management in the Christian Medical College, Vellore, India from July-September 2014 to estimate direct (drugs, tests, consultation/hospitalization) and indirect (travel, food, lost wages) costs associated with the episode. RESULTS: Median direct and indirect costs were Rs 590 and Rs 190 for out-patient management and Rs 7258 and Rs. 610 for hospitalization, constituting 1.1% and 11% of median annual household income, respectively. CONCLUSIONS: Escalating healthcare costs need tracking for evaluation of interventions.

The green tea component EGCG inhibits RNA polymerase III transcription.

RNA polymerase III (RNA pol III) transcribes many small structural RNA molecules involved in RNA processing and translation, and thus regulates the growth rate of a cell. Accurate initiation by RNA pol III requires the initiation factor TFIIIB. TFIIIB has been demonstrated to be regulated by tumor suppressors, including ARF, p53, RB, and the RB-related pocket proteins, and is a target of the oncogene c-myc and the mitogen-activated protein kinase ERK. EGCG has been demonstrated to inhibit the growth of a variety of cancer cells, induce apoptosis and regulate the expression of p53, myc, and ERK. Thus, we hypothesized that EGCG may regulate RNA pol III transcription in cells. Here, we report that EGCG (1) inhibits RNA pol III transcription from gene internal and gene external promoters (2) EGCG inhibits protein expression of the TFIIIB subunits Brf1 and Brf2, and (3) EGCG inhibits Brf2 promoter activity in cervical carcinoma cells.