A Dual Therapeutic Approach to Diabetes Mellitus via Bioactive Phytochemicals Found in a Poly Herbal Extract by Restoration of Favorable Gut Flora and Related Short-Chain Fatty Acids.

Diabetes mellitus (DM), a metabolic and endocrine condition, poses a serious threat to human health and longevity. The emerging role of gut microbiome associated with bioactive compounds has recently created a new hope for DM treatment. UHPLC-HRMS methods were used to identify these compounds in a poly herbal ethanolic extract (PHE). The effects of PHE on body weight (BW), fasting blood glucose (FBG) level, gut microbiota, fecal short-chain fatty acids (SCFAs) production, and the correlation between DM-related indices and gut microbes, in rats were investigated. Chebulic acid (0.368%), gallic acid (0.469%), andrographolide (1.304%), berberine (6.442%), and numerous polysaccharides were the most representative constituents in PHE. A more significant BW gain and a reduction in FBG level towards normal of PHE 600 mg/kg treated rats group were resulted at the end of 28th days of the study. Moreover, the composition of the gut microbiota corroborated the study's hypothesis, as evidenced by an increased ratio of Bacteroidetes to Firmicutes and some beneficial microbial species, including Prevotella copri and Lactobacillus hamster. The relative abundance of Bifidobacterium pseudolongum, Ruminococcus bromii, and Blautia producta was found to decline in PHE treatment groups as compared to diabetic group. The abundance of beneficial bacteria in PHE 600 mg/kg treatment group was concurrently associated with increased SCFAs concentrations of acetate and propionate (7.26 nmol/g and 4.13 nmol/g). The findings of this study suggest a promising approach to prevent DM by demonstrating that these naturally occurring compounds decreased FBG levels by increasing SCFAs content and SCFAs producing gut microbiota.

Phytochemicals, Antioxidant, Anti-inflammatory Studies, and Identification of Bioactive Compounds Using GC-MS of Ethanolic Novel Polyherbal Extract.

Hyperglycemia is the hallmark of diabetes, which is a collection of related metabolic disorders. Over time, diabetes can cause a variety of problems, including cardiovascular disease, nephropathy, neuropathy, and retinopathy. Ethanolic novel polyherbal extract (PHE) was prepared by mixing equal amounts of the following ingredients: Terminalia chebula Retz. (TC), Terminalia bellerica Roxb. (TB), Berberis aristata DC. (BA), Nyctanthes arbostratis L. (NA), Premna integrifolia L. (PI), and Andrographis paniculata Nees. (AP). Analysis of PHE results revealed phytochemicals like glycosides, flavonoids, alkaloids, tannins, phytosterols, and saponins. The aim of the study was to prepare an ethanolic extract of PHE using the cold maceration technique, and identify bioactive molecules from gas chromatography-mass spectrometry (GC-MS) analysis, and evaluate biological responses by using in vitro studies like antioxidant and anti-inflammatory activity. PHE was found to contain a total of 35 phytochemicals in GC-MS of which 22 bioactive compounds were obtained in good proportion. There are a few new ones, including 2-buten-1-ol, 2-ethyl-4-(2, 2, 3-trimethyl-3-cyclopenten-1-yl (17.22%), 1, 2, 5, 6-tetrahydrobenzonitrile (4.26%), 4-piperidinamine, 2, 2, 6, 6-tetramethyl-(0.07%), undecanoic acid, 5-chloro-, chloromethyl ester (0.41%), are identified. Antioxidant activity was estimated using EC50 values of 392.143 µg/ml, which were comparable to the standard value of EC50 310.513 µg/ml obtained using DPPH. Antioxidant activity was estimated with EC50 392.143 µg/ml, comparable to standard EC50 310.513 µg/ml using DPPH. In vitro anti-inflammatory potential was found with IC50 of 91.449 µg/ml, comparable to standard IC50 89.451 µg/ml for membrane stabilization and IC50 of 36.940 µg/ml, comparable to standard IC50 35.723 µg/ml for protein denaturation assays. As a result, the findings of this study show an enrichment of bioactive phytochemicals that can be used to investigate biological activity. To better understand how diabetes receptors work, in silico studies like docking could be carried out.

Residue Behavior of Clopyralid Herbicide in Soil and Sugar Beet Crop under Subtropical Field Conditions.

ABSTRACT: Sugar beet is a major crop for the sugar industry. With growing awareness of unsystematic use of pesticides, health problem, and environmental issues, assessment of pesticide residues in soil and crops has become necessary. Studies of subtropical conditions on dissipation and residue analysis of clopyralid have not yet been reported. Therefore, dissipation kinetics and terminal residues of clopyralid for two cropping seasons in the soil and the sugar beet crop were studied under field conditions. An experiment was laid out in a randomized block design, and a herbicide was applied as a postemergent. Clopyralid was extracted from the matrix by basic water, subjected to solid phase extraction cleanup, and quantified by high-pressure liquid chromatography-UV. The method was validated, and recovery percentage of pesticide ranged from 81 to 88, 77 to 85, 78 to 86, and 89 to 94% in the soil, sugar beet roots, sugar beet leaves, and water, respectively. After application in the soil, clopyralid dissipated rapidly following monophasic first-order kinetics, with a half-life of 13.39 days. Limits of detection and quantitation were 0.007 and 0.02 µg g-1, respectively. Clopyralid does not persist long in soil, and residues were below the European Union's maximum residue levels (0.5 mg kg-1) in the roots and leaves of sugar beet. Residues were also not detected in the groundwater. It can be concluded that clopyralid could be considered a safe herbicide from the environmental aspect due to its nonpersistence and that it would not have an adverse effect on human or animal health.

Inhibition of GSK-3 to induce cardiomyocyte proliferation: a recipe for in situ cardiac regeneration.

With an estimated 38 million current patients, heart failure (HF) is a leading cause of morbidity and mortality worldwide. Although the aetiology differs, HF is largely a disease of cardiomyocyte (CM) death or dysfunction. Due to the famously limited amount of regenerative capacity of the myocardium, the only viable option for advanced HF patients is cardiac transplantation; however, donor's hearts are in very short supply. Thus, novel regenerative strategies are urgently needed to reconstitute the injured hearts. Emerging data from our lab and others have elucidated that CM-specific deletion of glycogen synthase kinase (GSK)-3 family of kinases induces CM proliferation, and the degree of proliferation is amplified in the setting of cardiac stress. If this proliferation is sufficiently robust, one could induce meaningful regeneration without the need for delivering exogenous cells to the injured myocardium (i.e. cardiac regeneration in situ). Herein, we will discuss the emerging role of the GSK-3s in CM proliferation and differentiation, including their potential implications in cardiac regeneration. The underlying molecular interactions and cross-talk among signalling pathways will be discussed. We will also review the specificity and limitations of the available small molecule inhibitors targeting GSK-3 and their potential applications to stimulate the endogenous cardiac regenerative responses to repair the injured heart.

Prostaglandin-mediated inhibition of serotonin signaling controls the affective component of inflammatory pain.

Pain is fundamentally unpleasant and induces a negative affective state. The affective component of pain is mediated by circuits that are distinct from those mediating the sensory-discriminative component. Here, we have investigated the role of prostaglandins in the affective dimension of pain using a rodent pain assay based on conditioned place aversion to formalin injection, an inflammatory noxious stimulus. We found that place aversion induced by inflammatory pain depends on prostaglandin E2 that is synthesized by cyclooxygenase 2 in neural cells. Further, mice lacking the prostaglandin E2 receptor EP3 selectively on serotonergic cells or selectively in the area of the dorsal raphe nucleus failed to form an aversion to formalin-induced pain, as did mice lacking the serotonin transporter. Chemogenetic manipulations revealed that EP3 receptor activation elicited conditioned place aversion to pain via inhibition of serotonergic neurons. In contrast to their role in inflammatory pain aversion, EP3 receptors on serotonergic cells were dispensable for acute nociceptive behaviors and for aversion induced by thermal pain or a κ opioid receptor agonist. Collectively, our findings show that prostaglandin-mediated modulation of serotonergic transmission controls the affective component of inflammatory pain.

Posterior tibial nerve stimulation in the management of lower urinary tract symptoms in patients with multiple sclerosis.

INTRODUCTION AND HYPOTHESIS: Bladder dysfunction is a frequent symptom complex in patients with multiple sclerosis (MS) and often compromises the patient's quality of life. Pharmacotherapy has been poorly studied in the MS population showing contradictory results and reduced compliance owing to intolerable side effects. A new neuromodulation technique known as percutaneous tibial nerve stimulation (PTNS) has shown good efficacy and safety in the treatment of neurogenic and non-neurogenic lower urinary tract symptoms. In this article we review the literature and critically summarise the scientific evidence supporting the use of PTNS in the treatment of lower urinary tract symptoms (LUTS) in patients with MS. METHODS: We performed a computer-aided literature search in PubMed and EMBASE up to January 2015 to identify randomised controlled trials, case-control trials and prospective observational cohort studies. RESULTS: A total of 7 open-label, prospective studies and 313 MS patients were included. Three studies reported clinical and urodynamic outcomes at 3 to 9 months after PTNS; one study assessed the long-term efficacy of PTNS; two studies reported the acute effect of PTNS on urodynamic findings; one study assessed whether motor and sensory responses during PTNS could predict treatment outcome. None of the studies included a control group. CONCLUSIONS: Despite the very limited data PTNS seems an effective and safe treatment option in the management of LUTS in patients with MS.

Iron oxide nanoparticles and magnetic field exposure promote functional recovery by attenuating free radical-induced damage in rats with spinal cord transection.

BACKGROUND: Iron oxide nanoparticles (IONPs) can attenuate oxidative stress in a neutral pH environment in vitro. In combination with an external electromagnetic field, they can also facilitate axon regeneration. The present study demonstrates the in vivo potential of IONPs to recover functional deficits in rats with complete spinal cord injury. METHODS: The spinal cord was completely transected at the T11 vertebra in male albino Wistar rats. Iron oxide nanoparticle solution (25 µg/mL) embedded in 3% agarose gel was implanted at the site of transection, which was subsequently exposed to an electromagnetic field (50 Hz, 17.96 µT for two hours daily for five weeks). RESULTS: Locomotor and sensorimotor assessment as well as histological analysis demonstrated significant functional recovery and a reduction in lesion volume in rats with IONP implantation and exposure to an electromagnetic field. No collagenous scar was observed and IONPs were localized intracellularly in the immediate vicinity of the lesion. Further, in vitro experiments to explore the cytotoxic effects of IONPs showed no effect on cell survival. However, a significant decrease in H2O2-mediated oxidative stress was evident in the medium containing IONPs, indicating their free radical scavenging properties. CONCLUSION: These novel findings indicate a therapeutic role for IONPs in spinal cord injury and other neurodegenerative disorders mediated by reactive oxygen species.

Impacts of changing atmospheric deposition chemistry on nitrogen and phosphorus loading to Ganga River (India).

Investigations on atmospheric deposition (AD) and water chemistry along a 35 km stretch of Ganga River indicated that although N:P stoichiometry of AD did not change, there were over 1.4-2.0 fold increase in AD-NO3⁻, AD-NH4⁺ and AD-PO4³â» overtime. Concentration of dissolved inorganic-N (DIN) in river showed significant positive correlations with AD-NO3⁻ and runoff DIN. Similarly, dissolved reactive-P (DRP) in river showed significant positive correlation with AD-PO4³â» and runoff DRP. The study shows that AD has become an important source of N and P input to Ganga River.

Effect of thermal processing and γ-irradiation on allergenicity of legume proteins.

Legumes are implicated in IgE mediated food allergy in different countries. The present study aimed to investigate the effect of different processing methods on allergenicity of legume proteins. The extracts were processed by boiling, γ-irradiation or by combination of both. The changes in soluble protein content, specific IgE binding and allergenic potential of legume proteins were assessed. Thermal processing resulted in a 3- to 4-fold reduction in soluble protein. Specific IgE binding was reduced 74±6.5%, 83±11.6% and 62±7.2% in the soluble protein of kidney bean, black gram and peanut, respectively, after boiling (p<0.01) whereas there was 34±5.2%, 74±15.6% and 44±11.1% IgE binding reduction in the insoluble protein fraction of respective legumes. Boiling followed by γ-irradiation reduced IgE binding significantly (p<0.05). Biopotency of soluble protein of kidney bean, black gram and peanut was reduced 7-, 3- and 26-folds (p<0.001), respectively, and that of insoluble protein decreased 6-, 4- and 8-folds (p<0.001), respectively, after boiling. Combination treatment was effective in reducing the potency of both soluble and insoluble protein significantly as compared to boiling alone (p<0.001). However, γ-irradiation alone did not bring any change in allergenicity. In conclusion, boiling followed by γ-irradiation is effective in attenuating allergenicity of legume proteins.

Essential oil composition of Thymus serpyllum cultivated in the Kumaon region of western Himalaya, India.

The hydrodistilled essential oil of aerial parts of Thymus serpyllum L. (Lamiaceae), cultivated in the Kumaon region of western Himalaya was analyzed by GC and GC-MS. Twenty-nine compounds, representing 91.8% of the oil, were identified. The major components were thymol (58.8%), p-cymene (5.7%), thymol methyl ether (4.0%), borneol (3.8%), sabinene (3.4%), gamma-terpinene (3.4%) and carvacrol methyl ether (3.2%).

Soluble and nonsoluble protein assay for antigenic extracts from pollen and seeds of mustard (Brassica spp.).

A study on heterogeneity in water-soluble and non-water-soluble protein profiles of different species of Brassica pollen, seeds, and industrial flour for efficient allergy detection and immunotherapy has been lacking in India. The purpose of this study was to examine heterogeneity in the protein profile of antigenic extracts of different species of pollen, seeds, and seed flour of Brassica. A comparison of water-soluble and non-water-soluble protein profiles of seed extracts was studied. Water-soluble and non-water-soluble proteins were extracted, concentration was estimated by Lowry's method, and biochemical characterization by sodium dodecyl sulfate-polyacrylamide gel electrophoresis was analyzed. Pollen from four species, four species of Brassica seeds comprising 25 varieties, and industrial seed flour were included for water-soluble studies, whereas four species of seeds were included for non-water-soluble protein extraction. Significant variation in protein content was observed among four different species of pollen, seeds, and industrial seed flour, respectively. No significant variation was observed in non-water-soluble extracts of four species or among water-soluble and non-water-soluble content of seeds. Heterogeneity in the protein profile of different species of pollen was not observed. However, variation in banding pattern of water-soluble as well as non-water-soluble protein extracts among four different species of seeds was observed. Interestingly, the industrial seed flour also showed a rich protein banding pattern. Variation in protein content as well as protein profile among different species of seeds of Brassica is recorded.

Aeroallergens in clinical practice of allergy in India- ARIA Asia Pacific Workshop report.

Allergic diseases such as bronchial asthma, allergic rhinitis and atopic dermatitis are dramatically increasing all over the world including developing countries like India. Today, more than 30% of the population is known to suffer from one or other allergic ailment. Major causative agents implicated are pollen grains, fungal spores, dust mites, insect debris, animal epithelia, etc. Several aerobiological studies have been conducted in different parts of the country to ascertain aerial concentration and seasonality of pollen grains and fungi. Recently, an "All India Coordinated Project on Aeroallergens and Human Health" was undertaken by us to discover the quantitative and qualitative prevalence of aerosols at 18 different centers in the country. Allergenically important airborne pollen identified by clinico-immunologic evaluation are Alnus, Amaranthus, Argemone, Brassica, Cannabis, Cassia, Cedrus, Chenopodium, Cocos, Holoptelia, Mallotus, Morus, Parthenium, Prosopis juliflora, Quercus, Ricinus communis, and grasses such as Cenchrus, Cynodon, Imperata, Pennisetum etc. Cross-reactivity of the IgE antibodies is a common phenomenon among various pollen allergens. Ricinus communis pollen a commonly growing weed/shrub in India, cross-reacts with latex (Hevea brasiliensis), Mercurialis annua and also with seeds of Ricinus communis--all belonging to family Euphorbiaceae but geographically distantly located. Areca catechu cross-reacts with other members of Arecaceae such as Phoenix sylvestris, Cocos nucifera and Borassus flabelifer while pollen of Holoptelia integrifolia from India cross reacts with pollen of Parietaria judaica from Mediterranean Europe, both of which are members of family Urticaceae. Several reports on pollen and fruit syndrome have been analyzed. Experiments conducted by us revealed that pollutants (NO2 and SO2) not only affect pollen morphology but also changes its allergenic potency.

Protective effect of ethanolic and water extracts of sea buckthorn (Hippophae rhamnoides L.) against the toxic effects of mustard gas.

Ethanolic extract of H. rhamnoides L. leaf (HL-EOH), water and ethanolic extract of H. rhamnoides fruit (HF-W and HF-EOH), and H. rhamnoides flavone from fruit (HR-flavone) were evaluated against percutaneously administered sulphur mustard (SM), a chemical warfare agent. The animals administered with SM (9.7, 19.3 and 38.7 mg/kg) died at various days depending upon the dose and there was a significant reduction in the body weight. The H. rhamnoides extracts (1 g/kg; 3 doses; po) significantly protected the lethality, with a protective index of 2.4, 1.7, 1.7 and 2.2 for HL-EOH, HF-W, HF-EOH and HR-flavone respectively. Reduced glutathione (GSH) and oxidized glutalthione (GSSG) levels were reduced, and malondialdehyde (MDA) was elevated after percutaneous administration of SM. Oral administration of HL-EOH and HR-flavone significantly protected the body weight loss. Recovery in the levels of GSH, GSSG and MDA were also observed following oral administration of HL-EOH and HR-flavone. All the extracts were non-toxic and the LD50 was more than 5 g/kg. The present study shows that percutaneous administration of SM induces oxidative stress and ethanolic extract of leaf of H. rhamnoides and H. rhamnoides flavone from fruit can significantly protect it.

Algal meal supplementation of the cows' diet alters the physical properties of milk fat.

A three-week algal meal supplementation of the cows' basal diet resulted in an increase in the firmness of milk fat crystallized isothermally at 5 degrees C for 24 h--the apparent elastic constant increased from 100 to 224 N/mm. This was accompanied by a decrease in solid fat content, from 47.7% to 44.4%. The crystallization behaviour of milk fat was also modified significantly. The rate constant of crystallization (Avrami constant) of the enriched milk fat at 19 degrees C was approximatly 20 times higher than that of control milk fat. A shorter induction time of nucleation was also observed in the temperature range [20, 27 degrees C]. These effects were attributed to a higher degree of supersaturation of the enriched milk fat. Enriched milk fat nucleated in a more stable beta' polymorphic form at 5 degrees C, while control milk fat nucleated in the metastable alpha form, as determined by powder X-ray diffraction and differential scanning calorimetry. Changes in the microstructure of the material were observed by polarized light microscopy at 5 degrees C. The enriched milk fat displayed a greater amount of crystal clustering than the control. This effect was reflected in a decrease in the box-counting mass fractal dimension (Db) of the fat crystal network from 1.853 to 1.809. The decrease in Db closely predicted the observed 2.2-fold increase in the elastic constant of the fat. These changes in mechanical properties, crystallization behaviour and microstructure were driven by an increase in the 18:1 trans and a decrease in the 18:1 cis fatty acid content of the enriched milk fat.

Matching avrami indices achieves similar hardnesses in palm oil-based fats.

This study demonstrates how chemically interesterified hydrogenated palm oil (IHPO) and partially hydrogenated palm oil (PHPO) can be structured to have similar mechanical properties. Crystallization of IHPO at 30 degrees C for 24 h yielded a fat with a solid fat content (SFC) of 45% and a yield force of 51.5 N. On the other hand, PHPO had a SFC of 50% and a yield force of 44 N when crystallized under the same conditions. The result was opposite from what would be expected from the SFC point of view, thus suggesting that the microstructure of the fat plays a key role in determining mechanical properties. By matching crystallization behavior using the Avrami index as a guide, microstructures and material hardness were successfully matched. These results suggest that the dynamics of structure formation was a key factor influencing the macroscopic mechanical properties of palm oil-based fats.

Aeroallergens in clinical practice of allergy in India. An overview.

Allergic diseases such as bronchial asthma, allergic rhinitis and atopic dermatitis are dramatically increasing all over the world including developing countries like India. Today, more than 30 % of the population is known to suffer from one or other allergic ailment. Major causative agents implicated are pollen grains, fungal spores, dust mites, insect debris, animal epithelia, etc. Several aerobiological studies have been conducted in different parts of the country to ascertain aerial concentration and seasonality of pollen grains and fungi. Recently, an "All India Coordinated Project on Aeroallergens and Human Health" was undertaken to discover the quantitative and qualitative prevalence of aerosols at 18 different centres in the country. Predominant airborne pollen are Holoptelea, Poaceae, Asteraceae, Eucalyptus, Casuarina, Putanjiva, Cassia, Quercus, Cocos, Pinus, Cedrus, Ailanthus, Cheno/Amaranth, Cyperus, Argemone, Xanthium, Parthenium and others. Clinical and immunological evaluations have revealed allergenically important texa - some of them for the first time. Allergenically important pollen are Prosopis juliflora, Ricinus communis, Morus, Mallotus, Alnus, Querecus, Cedrus, Argemone, Amaranthus, Chenopodium, Holoptelea, Brassica, Cocos, Cannabis, Parthenium, Cassia and grasses. Further cross-reactivity of the IgE antibodies is a common phenomenon among various pollen allergens. Ricinus communis pollen from commonly growing weeds in India, cross-reacts with latex (Hevea brasiliensis), Mercurialis annua and also with seeds of Ricinus communis - all belonging to family Euphorbiaceae. Areca catechu cross-reacts with other members of Arecaceae such as Phoenix sylvestris, Cocos nucifera and Borassus flabelifer. Several reports on pollen and fruit syndrome have been analyzed. Experiments conducted by us revealed that pollutants (NO(2) and SO(2)) not only affect pollen morphology but also changes their allergenic potency. Immunotherapy with recombinant proteins having similar epitopes from different allergens have been advocated, besides allergen avoidance.

Antigenic and allergenic properties of Amaranthus Spinosus pollen--a commonly growing weed in India.

Amaranthus spinosus (Fam. Amaranthaceae) is an important aeroallergen in India and grows commonly in different parts of the country. In spite of its clinical significance in Type I hypersensitivity disorders, the antigenic and the allergenic properties of the pollen have not been systematically resolved. We investigated antigenic and allergenic properties of 5 pollen samples of Amaranthus spinosus collected from the Delhi area at fortnightly intervals. The protein content did not exhibit statistically significant variability. However, samples collected during the peak flowering season showed higher protein content. Biochemical characterization of samples showed multiple protein fractions by IEF and SDS-PAGE analysis. Samples collected during peak season showed a slightly higher number of bands (22) in the mw range of 14-70 kD. Seven protein fractions of 70, 66, 60, 50, 40, 30 and 14 kD were observed to have IgE binding capabilities and 9 were treated as allergenic. The observations will be helpful in standardizing pollen antigens for diagnosis and immunotherapy in India.

Allergen cross-reactivity between proteins of the latex from Hevea brasiliensis, seeds and pollen of Ricinus communis, and pollen of Mercurialis annua, members of the Euphorbiaceae family.

Allergen cross-reactions among three strongly sensitizing Euphorbiaceae species, i.e., the rubber tree (Hevea brasiliensis), castor bean (Ricinus communis), and the Mediterranean weed Mercurialis annua were studied in Finnish patients (n = 25) allergic to natural rubber latex (NRL), but with no known exposure to castor bean or M. annua, and French patients allergic to castor bean (n = 26) or to M. annua (n = 9), but not to NRL. In immunoglobulin E (IgE)-immunoblotting, 28% of NRL-allergic patient sera recognized castor bean seed and 48% reacted to castor bean pollen proteins. Likewise, 35% of the NRL-allergic patient sera bound to M. annua pollen allergens. Nineteen percent of castor bean-allergic patients showed IgE to NRL and 8% to M. annua proteins. Sera from patients allergic to M. annua reacted in 44% to NRL, in 56% to castor bean seed, and in 78% to castor bean pollen proteins. In immunoblotting, castor bean seed extract inhibited the binding of NRL-reactive IgE to 20 kDa, 30 kDa of NRL, and 55 kDa of proteins; NRL extract, in turn, inhibited the binding of castor bean-reactive IgE to 14, 21-22, 29, and 32-34 kDa of castor bean proteins. In ELISA inhibition, NRL extract inhibited 33% of the binding of M. annua--reactive IgE of pooled sera to M. annua pollen. In conclusion, allergen cross-reactivity in vitro was observed among three botanically related Euphorbiaceae members, H. brasiliensis, R. communis, and M. annua, but the molecular specificity of the observed cross-reactions as well as their clinical significance remains to be elucidated. Allergen cross-reactivity should be taken into account in diagnostic work.