Green synthesis of gold nanaoparticles using Delphinium Chitralense tuber extracts, their characterization and enzyme inhibitory potential.

Green synthesis has been introduced as an alternative to chemical synthesis due to the serious consequences. Metal nanoparticles synthesized through green approach have different pharmaceutical, medical and agricultural applications. The present study followed a green and simple route for the preparation of potentially bioactive gold nanoparticles (Au NPs). Au NPs were prepared via green synthesis approach using crude basic alkaloidal portion of the tuber of Delphinium chitralense. The green synthesized Au NPs were characterized by transmission electron microscopy (TEM), scanning electron microscopy (SEM), X-ray diffraction (XRD) fourier transform infrared (FTIR), and UV-Visible spectrophotometer. Morphological analysis shows that Au NPs have cubic geometry with different sizes. UV-Vis spectroscopic analysis confirmed the synthesis of Au NPs while XRD proved their pure crystalline phase. The Au NPs showed promising dose dependent inhibition of both AChE and BChE as compared to the crude as well as standard drug.

Effect of head covering on phototherapy induced hypocalcemia in term neonates with hyperbilirubinemia: A randomised controlled study.

BACKGROUND: Phototherapy is the primary treatment for hyperbilirubinemia in neonates. Hypocalcemia is a lesser known but potential detrimental effect of phototherapy. It has been hypothesized that phototherapy inhibits pineal secretion of melatonin, which blocks the effect of cortisol on bone calcium. Therefore, unchecked cortisol increases bone uptake of calcium and induces hypocalcemia. Covering head during phototherapy in order to prevent light reaching to the pineal gland which eventually leads to the prevention of hypocalcemia is hypothesized to prevent hypocalcemia but it lacks sufficient evidence worldwide. METHOD: It is a prospective, randomized controlled study. 112 neonates were randomized into two groups of 56 neonates. Group A underwent phototherapy without head cover and group B with head covered by a cap. RESULT: The mean decline in serum ionic calcium after 48 hours of phototherapy in group A and group B was 0.57±0.37 mg/dl and 0.34±0.24 mg/dl respectively. This decline in serum ionic calcium was significantly higher in group A. (p < 0.001). 26.8% newborns from group A developed hypocalcemia while in group B only 14.3% developed hypocalcemia however it was not found to be statistically significant. Incidence of symptomatic hypocalcemia between the two groups was also not significant. CONCLUSION: There was significant reduction in serum calcium in neonates undergoing phototherapy without head cover as compared to neonates with head cover but risk of hypocalcemia was not significant. Further studies with larger sample size including preterm are recommended.

Inflammation and oxidative stress transcription profiles due to in vitro supply of methionine with or without choline in unstimulated blood polymorphonuclear leukocytes from lactating Holstein cows.

Neutrophils are the most important polymorphonuclear leukocytes (PMNL), representing the front-line defense involved in pathogen clearance upon invasion. As such, they play a pivotal role in immune and inflammatory responses. Isolated PMNL from 5 mid-lactating Holstein dairy cows were used to evaluate the in vitro effect of methionine (Met) and choline (Chol) supplementation on mRNA expression of genes related to the Met cycle and innate immunity. The target genes are associated with the Met cycle, cell signaling, inflammation, antimicrobial and killing mechanisms, and pathogen recognition. Treatments were allocated in a 3 × 3 factorial arrangement, including 3 Lys-to-Met ratios (L:M, 3.6:1, 2.9:1, or 2.4:1) and 3 levels of supplemental Chol (0, 400, or 800 µg/mL). Three replicates per treatment group were incubated for 2 h at 37°C and 5% atmospheric CO2. Both betaine-homocysteine S-methyltransferase and choline dehydrogenase were undetectable, indicating that PMNL (at least in vitro) cannot generate Met from Chol through the betaine pathway. The PMNL incubated without Chol experienced a specific state of inflammatory mediation [greater interleukin-1ß (IL1B), myeloperoxidase (MPO), IL10, and IL6] and oxidative stress [greater cysteine sulfinic acid decarboxylase (CSAD), cystathionine gamma-lyase (CTH), glutathione reductase (GSR), and glutathione synthase (GSS)]. However, data from the interaction L:M × Chol indicated that this negative state could be overcome by supplementing additional Met. This was reflected in the upregulation of methionine synthase (MTR) and toll-like receptor 2 (TLR2); that is, pathogen detection ability. At the lowest level of supplemental Chol, Met downregulated GSS, GSR, IL1B, and IL6, suggesting it could reduce cellular inflammation and enhance antioxidant status. At 400 µg/mL Chol, supplemental Met upregulated PMNL recognition capacity [higher TLR4 and L-selectin (SELL)]. Overall, enhancing the supply of methyl donors to isolated unstimulated PMNL from mid-lactating dairy cows leads to a low level of PMNL activation and upregulates a cytoprotective mechanism against oxidative stress. Enhancing the supply of Met coupled with adequate Chol levels enhances the gene expression of PMNL pathogen-recognition mechanism. These data suggest that Chol supply to PMNL exposed to low levels of Met effectively downregulated the entire repertoire of innate inflammatory-responsive genes. Thus, Met availability in PMNL during an inflammatory challenge may be sufficient for mounting an appropriate biologic response.

Alpha lipoic acid supplementation ameliorates the wrath of simulated tropical heat and humidity stress in male Murrah buffaloes.

A supplement which ameliorates temperature-humidity menace in food producing livestock is a prerequisite to develop climate smart agricultural packages. A study was conducted to investigate the heat stress ameliorative efficacy of alpha lipoic acid (ALA) in male Murrah water buffaloes (Bubalus bubalis). Eighteen animals (293.61 ± 4.66Kg Bwt) were randomly allocated into three groups (n = 6); NHSC (non-heat-stressed control), HS (heat-stressed) and HSLA (heat-stressed-supplemented with ALA@32 mg/kg Bwt orally) based on the temperature humidity index (THI) and ALA supplementation. HS and HSLA were exposed to simulated heat challenge in a climatically controlled chamber (40 °C) for 21 consecutive days, 6 h daily. Physiological responses viz. Respiration rate (RR), Pulse rate (PR) and Rectal temperature (RT) were recorded daily before and after heat exposure. Blood samples were collected at the end of heat exposure on days 1, 6, 11, 16, and 21 and on day 28 (7th day post exposure which is considered as recovery) for peripheral blood mononuclear cells (PBMCs) separation, followed by RNA and Protein extraction for Real time quantitative PCR and Western blot analysis respectively, of heat shock proteins (HSPs). Two-way repeated measure ANOVA was performed between groups at different experimental periods. RR (post exposure) in HS and HSLA was significantly higher (P < 0.05) than NHSC from day 1 onwards but HSLA varied significantly from the HS 8th day onwards. Post exposure RT and PR in both HS and HSLA varied (P < 0.05) from NHSC throughout the study; but between HS and HSLA, RT significantly varied on initial 2 days and last 6 days (from days 16 to 21). HSP70 mRNA expression significantly up regulated in high THI groups with respect to the low THI group throughout the experimental period. During chronic stress (days 16 and 21) HSP70 significantly (P < 0.05) increased in HS but not in HSLA (P > 0.05) with respect to NHSC. ALA supplementation up-regulates and sustains (P < 0.05) the expression of HSP90 in HSLA in comparison to the HS and NHSC. HSP105 expression was significantly up-regulated (P < 0.05) in HS on days 16 and 21 (during long-term exposure) but only on day 21 (P < 0.05) in HSLA. HSP70, HSP90, and HSP105 protein expression dynamics were akin to the mRNA transcript data between the study groups. In conclusion, supplementing ALA ameliorates the deleterious effect of heat stress as reflected by improved physiological and cellular responses. ALA supplementation improved cellular antioxidant status and sustained otherwise easily decaying heat shock responses which concertedly hasten the baton change from a limited window of thermo tolerance to long run acclimatization.

In vivo and in vitro Pharmacological Evaluation of Gymnosporia Royleana / Evaluación Farmacológica de Gymnosporia Royleana in vivo e in vitro

ABSTRACT Objective: Gymnosporia royleana (G royleana) Wall ex MA Lawson, locally known as (Sur Azghee), is traditionally used for the management of various diseases. In the current investigation, we made an effort to scientifically validate its traditional use in various pathological conditions, such as microbial infections and cancer, and to explore its additional pharmacological activities via random screening against locally accessible pharmacological methods, irrespective of its traditional uses like antidiabetic, haemagglutination and antioxidant assays. Methods: Extraction was carried out using a cold maceration methodology. Dilution method was used for antimicrobial susceptibility testing using different concentrations. Streptozocin (STZ) induced protocol was used to assess antidiabetic activity at a dose level of 200, 400 mg/ kg. Antioxidant activity, haemagglutination activity, and anticancer activities against HepG-2 and MCF-7 cell lines were determined as per established protocols. Similarly, the maximum amount of phenolic content (12.02 mg 100 g) was determined by using Folin Ciocalteu assay. Results: Promising antimicrobial activities in terms of minimum inhibitory concentration (MIC) were noted for crude extract (25-200 µg/mL), n-hexane (100-400 µg/mL), ethyl acetate (50-200 µg/mL) and aqueous (100-400 µg/mL). Antidiabetic potential was significant at a dose level of 200-400 mg/kg bodyweight by reducing the blood glucose level at days 10 and 15. The percentage of 2,2-diphenyl-1-picrylhydrazyl (DPPH) values increase by increasing the concentration of the plant extract (10-100 µg/mL). The methanol extract was found to possess high agglutination activity. Conclusion: It was concluded that this plant species possess significant antimicrobial, antidiabetic, antioxidant, anticancer and haemagglutination activities, which could be attributed to the phenolic content of the extract.

RESUMEN Objetivo: Gymnosporia royleana (G royleana) Wall ex MA Lawson, localmente conocida como "Sur Azghee", se utiliza tradicionalmente para el tratamiento de diversas enfermedades. En la investigación presente, tratamos de validar científicamente su uso tradicional en varias condiciones, tales como las infecciones microbianas y el cáncer, así como explorar sus actividades farmacológicas adicionales mediante el tamizado aleatorio frente a los métodos farmacológicos localmente accesibles, independientemente de sus usos tradicionales como ensayos antidiabéticos, hemaglutinantes, y antioxidantes. Métodos: La extracción se realizó mediante una metodología de maceración en frío. Un método de dilución se utilizó para la prueba de susceptibilidad antimicrobiana utilizando diferentes concentraciones. Se utilizó el protocolo inducido por estreptozotocina (STZ) para evaluar la actividad antidiabética a un nivel de dosis de 200, 400 mg/kg. La actividad antioxidante, la actividad de hemaglutinación, y las actividades anticancerígenas contra las líneas celulares HepG-2 y MCF-7, se determinaron según los protocolos establecidos. De modo similar. la cantidad máxima de contenido fenólico (12.02 mg 100 g) se determinó mediante el uso del ensayo Folin-Ciocalteu. Resultados: Se observaron actividades antimicrobianas prometedoras en términos de la concentración inhibitoria mínima (CIM) para el extracto crudo (25-200 μg/mL), el n-hexano (100-400 μg/mL), el acetato de etilo (50-200 μg/mL), y el extracto acuoso (100-400 μg/mL). El potencial antidiabético fue significativo a un nivel de dosis de 200-400 mg/kg de peso corporal mediante la reducción del nivel de glucosa en sangre a los 10 y 15 días. El porcentaje de los valores de 2,2-difenil-1-picrilhidracilo (DPPH) se incrementa al aumentar la concentración del extracto de la planta (10-100 μg/mL). Se halló que el extracto de metanol posee una alta actividad de aglutinación. Conclusión: Se concluyó que esta especie de planta posee importantes actividades antimicrobianas, antidiabéticas, antioxidantes, anticancerígenas y hemaglutinantes, que podrían atribuirse al contenido fenólico del extracto.

Antibacterial and Antifungal Activities of Compounds Isolated from Gymnosporia Royleana (Celastraceae) / Actividades antibacterianas y antifúngicas de compuestos aislados de Gymnosporia Royleana (Celastraceae)

ABSTRACT Objective: In the present study, five new source compounds isolated from aerial parts of Gymnosporia royleana (G royleana) were screened for antibacterial and antifungal activities. Methods: Extraction from plant material was carried out using cold maceration technique. Isolation of pure compounds was accomplished through repeated column chromatography of different fractions obtained from crude extract and using silica gel as stationary phase. Their structures were established via advanced spectroscopic techniques along with the spectral data previously reported for these compounds. Dilution method was used for the evaluation of antimicrobial potential of the compounds against various microbial strains. Results: Among the tested compounds, Gymnosporin B displayed moderate antimicrobial activity against Escherichia Coli (E coli), Staphylococcus aureus (S aureus), Candida albicans (C albicans) and Aspergillus flavus (A flavus) [minimum inhibitory concentration (MIC) range; 32-64 μg/mL]. Similarly, Gymnosporin C also showed moderate activity against E coli and S aureus (MIC; 32 μg/mL each) as well as weak activity against C albicans and A flavus (MIC; 64 μg/mL each). However, Royaflavone showed moderate antibacterial activity against S aureus only (MIC; 32 μg/mL). Antimicrobial activity of the rest of the compounds was weak and negligible. Conclusion: The present study has provided fascinating results of antimicrobial activities of the isolated compounds. However, the broad antimicrobial spectrum of Gymnosporin B and Gymnosporin C demands for further exploration of these triterpenes, both on the basis of mechanism and quantitative structure-activity relationship.

RESUMEN Objetivo: En el presente estudio, cinco nuevos compuestos de origen aislados de partes aéreas de Gymnosporia royleana (G royleana) fueron tamizados en sus actividades antibacterianas y antifúngicas. Métodos: La extracción de material vegetal se realizó mediante la técnica de maceración en frío. El aislamiento de compuestos puros se logró a través de la cromatografía en columna repetida de diversas fracciones obtenidas del extracto crudo y usando gel de silicona como fase estacionaria. Sus estructuras fueron establecidas mediante técnicas espectroscópicas avanzadas junto con los datos espectrales previamente reportados para estos compuestos. El método de dilución fue usado para evaluar el potencial antimicrobiano de los compuestos contra diversas cepas microbianas. Resultados: Entre los compuestos sometidos a prueba, Gymnosporina B mostró una actividad antimicrobiana moderada contra Escherichia Coli (E coli), Staphylococcus aureus (S aureus), Candida Albicans (C albicans) y Aspergillus flavus (A flavus) [rango de concentración inhibitoria mínima (CIM); 32 - 64 μg/mL]. De manera similar, Gymnosporina C también mostró actividad moderada contra E coli y S aureus (CIM; 32 μg/mL cada uno) así como débil actividad frente a C albicans y A flavus (CIM; 64 μg/mL cada uno). Sin embargo, Royaflavone mostró actividad antibacteriana moderada sólo frente a S aureus (CIM; 32 μg/mL). La actividad antimicrobiana del resto de los compuestos fue débil e insignificante. Conclusión: El presente estudio ha proporcionado resultados interesantes acerca de las actividades antimicrobianas de los compuestos aislados. Sin embargo, el amplio espectro antimicrobiano de la Gymnosporina B y la Gymnosporina C exige una mayor exploración de estos triterpenos, tanto sobre la base del mecanismo como a partir de la relación cuantitativa estructura-actividad.

In-vivo Assessment of Antipyretic and Anti-inflammatory Activities of Gymnosporia Royleana Extract in Mice / Evaluación in vivo de las Actividades Antipiréticas y Antiinflamatorias del Extracto de Gymnosporia Royleana en Ratones

ABSTRACT Objective: Despite the presence of multitude of synthetic drugs against fever and inflammation, none has been proven entirely safe. In contrast, the accepted safety of plant derived natural products is inspiring the world. Based on this fact as well as in view of the diversified activities reported from the genus Gymnosporia, the present study was designed to evaluate the antipyretic and anti-inflammatory activity of Gymnosporia royleana (G royleana). Methods: The methanolic extract of the aerial parts of G royleana was screened for in-vivo antipyretic activity using the brewer's yeast-induced pyrexia mice model and for anti-inflammatory activity using the carrageenan-induced paw oedema and xylene-induced ear oedema mice model. Results: In the antipyretic assay, G royleana extract showed considerable antipyretic activity in a dose dependent fashion. Statistically significant antipyretic effects (p < 0.05) were observed at the end of the second hour of administration for all doses of extract and remained significant until the end of the experiment. The plant extract also displayed promising anti-inflammatory activity, in a dose dependent fashion, in both models of inflammation ie carrageenan- and xylene-induced oedema models, when compared to the controls. In the carrageenan-induced oedema model, significant effects (p < 0.01) were observed for 300 and 600 mg/kg doses after 60 minutes of xylene administration (ie 55.51% and 65.88% inhibition of oedema, respectively). Conclusion: The study provided evidence supporting the antipyretic and anti-inflammatory activity of the G royleana methanolic extract.

RESUMEN Objetivo: A pesar de la presencia de multitud de fármacos sintéticos en el arsenal contra la fiebre y la inflamación, ninguno ha dado pruebas de ser completamente seguro. En contraste con ello, la seguridad aceptada de los productos naturales derivados de las plantas inspira al mundo. Sobre la base de este hecho, así como en vista de las actividades diversificadas que se reportan con respecto al género Gymnosporia, el presente estudio se diseñó con el objeto de evaluar el potencial antipirético y antiinflamatorio de Gymnosporia royleana (G royleana). Métodos: El extracto de metanol de las partes aéreas de G royleana fue tamizado en busca de actividad antipirética in vivo, utilizando el modelo de pirexia inducida por levadura de cerveza en ratones, y de actividad antiinflamatoria utilizando modelos de ratones con oedema de las patas inducido mediante carragenina, y oedema de las orejas inducido mediante xileno. Resultados: En el ensayo antipirético, el extracto de G royleana mostró una actividad antipirética considerable en forma dependiente de la dosis. Se observó un efecto antipirético estadísticamente significativo (p < 0.05) en el transcurso de la segunda hora de administración para todas las dosis de extracto y se mantuvo significativo hasta el final del experimento. El extracto de la planta también mostró una actividad antiinflamatoria prometedora, de una manera dependiente de la dosis, en ambos modelos de inflamación, es decir, modelos de oedema inducido por carragenina y xileno, en comparación con el control. En el modelo de oedema inducido por carragenina, se observó un efecto significativo (p < 0.01) para dosis de 300 y 600 mg / kg después de 60 minutos de administración de xileno (es decir, 55.51% y 65.88% de inhibición del oedema, respectivamente). Conclusión: El estudio proporcionó pruebas suficientes sobre el potencial antipirético y antiinflamatorio del extracto de G royleana.

Sub-ppm level high energy resolution fluorescence detected X-ray absorption spectroscopy of selenium in articular cartilage.

The speciation of highly-diluted elements by X-ray absorption spectroscopy in a diverse range of materials is extremely challenging, especially in biological matrices such as articular cartilage. Here we show that using a high energy resolution fluorescence detected X-ray absorption spectroscopy (HERFD-XAS) technique coupled to an array of crystal analyzers, selenium speciation down to 400 ppb (µg kg-1) within articular cartilage can be demonstrated. This is a major advance in the speciation of highly-diluted elements through X-ray absorption spectroscopy and opens new possibilities to study the metabolic role of selenium and other elements in biological samples.

Cryopreservation of Axillary Buds of Cannabis sativa L. by V-Cryoplate Droplet-Vitrification: The Critical Role of Sucrose Preculture.

BACKGROUND: Cryopreservation is the only method allowing the safe and cost-effective long-term conservation of important germplasm. Recent use of the cryo-plate system has proven beneficial in further simplifying the cryopreservation protocols. OBJECTIVE: Developing an efficient protocol for the cryopreservation of axillary buds of Cannabis sativa elite cultivars (MX and V1-20) by the V-cryoplate droplet-vitrification technique. MATERIALS AND METHODS: Stem segments (~5 cm in length) with mature axillary buds collected from indoor-grown plants were surface sterilized and then either precultured on MS basal medium with 0.1 M sucrose (1st step preculture) for 72 h or non-precultured. All mature axillary buds (~1 mm) were aseptically excised from stem segments and precultured for an additional 48 h on MS basal medium with sucrose (0.3 M) and 5% DMSO prior to cryopreservation (2nd step preculture). Biomass samples of fully mature mother plants and regrown cryopreserved plants were analyzed for Δ9-THC and CBD content using gas chromatography-flame ionization detector (GC/FID). RESULTS: The survival and regrowth rates of cryopreserved axillary buds of cultivar MX following this two-step preculture were 45% and 42% respectively, while those of cultivar V1-20 were 47% and 44% respectively. A direct preculture of axillary buds (2nd step preculture) on high sucrose (0.3M sucrose) significantly decreased both the survival and regrowth levels of axillary buds of cultivar MX (5% and 3% respectively) as well as those of cultivar V1-20 (20% and 17% respectively). Δ9-THC and CBD content of mother plants and regrown cryopreserved plants were found to be highly comparable to each other. CONCLUSION: The resulting plants after cryopreservation appeared normal without any callus formation or morphogenetic variation. On maturity, mother plants and re-grown cryopreserved plants were comparable in terms of Δ9-THC and CBD content. This report provides an efficient protocol for cryopreservation of axillary buds of Cannabis sativa cultivars which may be applicable to other important cultivars, plant parts and other related medicinal plants.

Effect of Moringa oleifera leaf powder supplementation on growth performance and intestinal morphology in broiler chickens.

Due to public health hazards of subtherapeutic use of antibiotics in poultry feed, there is a need for alternatives. Moringa oleifera is a phytobiotic, which is reported to possess antimicrobial and immuno-modulatory properties. This study aimed to investigate effects of M. oleifera leaf powder (MOLP) supplementation on growth performance and intestinal microarchitecture of broiler chickens. Day-old broiler chicks (n = 100) were randomly divided into five groups with four replicates each having five birds. Birds were fed a corn-based basal diet or the same diet supplemented with 0.6, 0.9, 1.2 and 1.5% MOLP. Body weight and feed conversion ratio were recorded on weekly basis. On day 35, two birds per replicate were selected to collect samples of duodenum, jejunum, ileum, caecal tonsils and bursa of Fabricius. Tissue samples were stained with haematoxylin and eosin or combined alcian blue and PAS technique. Supplementation of MOLP had no effect on feed intake, feed conversion ratio, bursa weight and intra-epithelial lymphocytes (IEL) count. Birds fed with 1.2% MOLP had higher (p < 0.05) body weight, length and empty weight of small intestine, and weight of empty and filled ceca. Villus height (duodenum, jejunum, ileum), villus surface area (duodenum) and villus height:crypt depth (ileum) were higher (p < 0.05) in 1.2% MOLP group compared with non-supplemented group. In duodenum, total goblet cell count was higher (p < 0.05) in all the supplemented groups when compared with non-supplemented birds. Acidic mucin count was higher (p < 0.05) in duodenum, jejunum and ileum of 1.2% MOLP group compared with the control group. Bursal follicle count was higher (p < 0.05) in 1.2% MOLP group compared with non-supplemented group. In conclusion, dietary supplementation of 1.2% MOLP modulated intestinal microarchitecture and acidic mucin production without any apparent effect on growth.

Laser-activated transforming growth factor-ß1 induces human ß-defensin 2: implications for laser therapies for periodontitis and peri-implantitis.

BACKGROUND: There is increasing popularity of high-power lasers for surgical debridement and antimicrobial therapy in the management of peri-implantitis and periodontal therapy. Removal of the noxious foci would naturally promote tissue healing directly. However, there are also anecdotal reports of better healing around routine high-power laser procedures. The precise mechanisms mediating these effects remain to be fully elucidated. This work examines these low-dose laser bystander effects on oral human epithelial and fibroblasts, particularly focusing on the role of human ß-defensin 2 (HBD-2 or DEFB4A), a potent factor capable of antimicrobial effects and promoting wound healing. MATERIAL AND METHODS: Laser treatments were performed using a near-infrared laser (810 nm diode) at low doses. Normal human oral keratinocytes and fibroblast cells were used and HBD-2 mRNA and protein expression was assessed with real time polymerase chain reaction, western blotting and immunostaining. Role of transforming growth factor (TGF)-ß1 signaling in this process was dissected using pathway-specific small molecule inhibitors. RESULTS: We observed laser treatments robustly induced HBD-2 expression in an oral fibroblast cell line compared to a keratinocyte cell line. Low-dose laser treatments results in activation of the TGF-ß1 pathway that mediated HBD-2 expression. The two arms of TGF-ß1 signaling, Smad and non-Smad are involved in laser-mediated HBD-2 expression. CONCLUSIONS: Laser-activated TGF-ß1 signaling and induced expression of HBD-2, both of which are individually capable of promoting healing in tissues adjacent to high-power surgical laser applications. Moreover, the use of low-dose laser therapy itself can provide additional therapeutic benefits for effective clinical management of periodontal or peri-implant disease.

Chemical composition and biological activities of Artemisia judaica essential oil from southern desert of Jordan.

ETHNOPHARMACOLOGIC RELEVANCE: Artemisia judaica L. (Arabic name: Beithran), is a medicinal and aromatic plant growing in the valley bottoms of desert areas, particularly in the southern desert of Jordan nearest to the Jordan-Saudi Arabia borders and in Wadi Araba in the Southern Badia. In Jordan, A. judaica is widely used in traditional medicine being recommended by aboriginal Bedouins in the North Badia region of Jordan as calmative. Furthermore, it is used for the treatment of stomach ache, heart diseases, sexual weakness, diabetes, gastro-intestinal disorders and external wounding. Additionally, other folk medicines of the Arabic region commonly use this aromatic plant for the treatment of inflammatory-related diseases, for instance fungal infections, diabetes, atherosclerosis, cancer and arthritis. AIM OF THE STUDY: Considering the traditional medicinal uses and the lack of scientific studies addressing the cellular and molecular mechanisms behind A. judaica claimed activities, the present study was designed to validate some of the traditional uses ascribed to this species, specifically the antifungal and anti-inflammatory activities of A. judaica essential oil at doses devoid of cytotoxicity to mammalian cells. MATERIALS AND METHODS: Chemical analysis of A. judaica essential oil isolated by hydrodistillation from aerial parts was carried out by gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS). The antifungal activity (minimal inhibitory concentrations and minimal lethal concentrations) was evaluated against yeasts, dermatophyte and Aspergillus strains. In order to deeply explore the mechanisms behind the anti-fungal effect of the essential oil, the germ tube inhibition assay and the biofilms formation assay were evaluated using Candida albicans. The assessment of cell viability was accomplished using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay in both hepatocytes and macrophages. Furthermore, the in vitro anti-inflammatory potential of A. judaica oil was evaluated by measuring nitric oxide (NO) production using lipopolysaccharide (LPS)-stimulated mouse macrophages. RESULTS: Oxygen containing monoterpenes are a representative group of constituents (68.7%) with piperitone (30.4%), camphor (16.1%) and ethyl cinnamate (11.0%) as main compounds. The highest antifungal activity of the oil was observed against Cryptococcus neoformans, with a MIC value of 0.16µL/mL. The oil revealed an important inhibitory effect on germ tube formation in C. albicans with 80% inhibition of filamentation at a concentration of 0.16µL/mL. Importantly, the oil also interfered with pre-formed biofilms by reducing the amount of the attached biomass. Furthermore, the essential oil significantly inhibited NO production evoked by LPS on macrophages at concentrations with very low toxicity (0.32µL/mL) or without toxicity (0.16µL/mL) to both macrophages and hepatocytes. CONCLUSIONS: The present study revealed that A. judaica essential oil from Jordan significantly inhibited germ tube formation and disrupted preformed biofilms of C. albicans, emphasizing the therapeutic potential for the treatment of disseminated candidiasis. Additionally, safe concentrations of this essential oil significantly inhibited NO production elicited by LPS in macrophages, highlighting its potential anti-inflammatory activity. Overall, A. judaica bears promising therapeutic potential for further drug development. Importantly, this work also validates some of the traditional uses of A. judaica.

Modulation of ethanol toxicity by Asian ginseng (Panax ginseng) in Japanese ricefish (Oryzias latipes) embryogenesis.

Alcohol consumption by women during pregnancy often induces fetal alcohol spectrum disorder (FASD) in children who have serious central nervous system (CNS), cardiovascular, and craniofacial defects. Prevention of FASD, other than women abstaining from alcohol drinking during pregnancy, is not known. A limitation of the use of synthetic anti-alcoholic drugs during pregnancy led us to investigate herbal products. In particular, many plants including Asian ginseng (Panax ginseng) have therapeutic potential for the treatment of alcoholism. We used Japanese ricefish (medaka) (Oryzias latipes), an animal model of FASD, for identifying herbal medicines that can attenuate ethanol toxicity. Fertilized eggs in standard laboratory conditions were exposed to ginseng (PG) root extract (0-2 mg/mL) either 0-2 (group A) or 1-3 (group B) day post fertilization (dpf) followed by maintenance in a clean hatching solution. The calculated IC50 as determined 10 dpf in A and B groups were 355.3±1.12 and 679.7±1.6 µg/mL, respectively. Simultaneous exposure of embryos in sub-lethal concentrations of PG (50-200 µg/mL) and ethanol (300 mM) for 48 h disrupted vessel circulation and enhanced mortality. However, PG (100 µg/mL) may partially protect trabecular cartilage (TC) deformities in the neurocranium in B group embryos induced by ethanol (300 mM). To understand the mechanism, embryonic ethanol concentration was measured at 2 dpf and adh5, adh8, aldh2, aldh9a, catalase, GST, and GR mRNAs were analyzed at 6 dpf. It was observed that although ethanol is able to reduce adh8 and GST mRNA contents, the simultaneous addition of PG was unable to alter ethanol level as well as mRNA contents in these embryos. Therefore, antagonistic effects of PG on ethanol toxicity are mediated by a mechanism which is different from those regulating ethanol metabolism and oxidative stress.

Preclinical profile of bacopasides from Bacopa monnieri (BM) as an emerging class of therapeutics for management of chronic pains.

Chronic pains management costs billions of dollars in medical exchequer to the world population. Additionally, 77% of people with chronic pains also have a degree of medically treatable depression. Opioids have a narrower safety index due to their side effects associated with its tolerance, hyperalgesia and subsequent dependence. Likewise, non steroidal anti-inflammatory drugs and anticonvulsants, also have limited safety and tolerability profile in the management of chronic pains. Bacopa monnieri, a renowned ayurvedic medicine has a strong antidepressant effect and significant antinociceptive effect, which is comparable to the effect of morphine via adenosinergic, opioidergic, and adrenergic mechanisms. BM has been also reported to be effective in neuropathic pains. Additionally, it has a strong anti-inflammatory effect mediated via COX-2 inhibitory mechanism. Apart from its effect of augmenting morphine analgesia, BM also inhibits opioid-withdrawal induced hyperalgesia, and acquisition and expression of morphine tolerance. BM is reported to have a strong protective effect against toxic effects of opiates on major organs like brain, kidneys and heart. BM is well documented to be safe and well tolerated herbal therapy in multiple clinical trials including various age groups. This minireview evaluated the preclinical data that highlights potential of BM as a future candidate for clinical management of chronic pains.

4-epi-Pimaric acid: a phytomolecule as a potent antibacterial and anti-biofilm agent for oral cavity pathogens.

The present study focused on the antibacterial and biofilm inhibitory potential of 4-epi-pimaric acid isolated from aerial parts (stem and leaves) of Aralia cachemirica L. (Araliaceae) against oral cavity pathogens. 4-epi-Pimaric acid exhibited minimum inhibitory concentration (MIC) in the range of 4-16 µg/ml and minimum bactericidal concentration (MBC) two- to four-folds higher than MIC. There was significant inhibition in the biofilm formation by Streptococcus mutans on the saliva coated surface (P < 0.05), and confocal microscopy revealed that 4-epi-pimaric acid inhibited the clumping and attachment of S. mutans. At 8 × MIC concentration, it significantly prevented the pH drop and reduced S. mutans biofilms (P < 0.05). Increased propidium iodide staining and leakage of 260- and 280-nm absorbing material by 4-epi-pimaric acid treated cells of S. mutans suggested that it probably causes disruption of the cytoplasmic membrane structure. It also exhibited significant suppression of TNF-α expression in human neutrophils, suggestive of its anti-inflammatory activity. Furthermore, the compound was found to be significantly safe (IC(50) >100 µg/ml) in the MTT assay on AML-12 cell lines. In conclusion, 4-epi-pimaric acid showed promising antibacterial, anti-biofilm and anti-inflammatory potency and this compound can be exploited for therapeutic application in oral microbial infections.

Wear and surface analysis of 38 mm ceramic-on-metal total hip replacements under standard and severe wear testing conditions.

The purpose of this study was to compare the wear of zirconia-toughened alumina (ZTA) and alumina femoral heads tested against as-cast CoCrMo alloy acetabular cups under both standard and severe wear conditions. A new severe test, which included medio-lateral displacement of the head and rim impact upon relocation, was developed. This resulted in an area of metal transfer and an area of increased wear on the superior-anterior segment of the head that were thought to be due to dislocation and rim impact respectively. While the wear of all ceramic heads was immeasurable using the gravimetric method, the wear rates for the metallic cups from each test were readily calculated. An average steady state wear rate of 0.023 +/- 0.005 mm3/10(6) cycles was found for the cups articulating against ZTA under standard wear conditions. A similar result had previously been obtained for the wear of cups articulated against alumina heads of the same size (within the same laboratory). Under severe wear conditions an increase in the metallic cup steady state wear rate was found with the ZTA and alumina tests giving 0.623 +/- 0.252 and 1.35 +/- 0.154 mm3/10(6) cycles respectively. Wear of the ceramic heads was detected using atomic force microscopy which showed, under severe wear conditions, a decrease in polishing marks and occasional grain removal. The surfaces of the ZTA heads tested under standard conditions were virtually unchanged from the unworn samples. Friction tests showed low friction factors for all components, pre and post wear.

In vitro antioxidant and radical-scavenging capacities of Citrullus colocynthes (L) and Artemisia absinthium extracts using promethazine hydrochloride radical cation and contemporary assays.

A new, quick and economical decolorization assay based upon the generation of a radical cation made from promethazine hydrochloride (PMZH) is described for screening of antioxidant activity of plants/herbal extracts. PMZH radical cations, produced through a reaction between PMZH and potassium persulfate (K(2)S(2)O(8)) in phosphoric acid medium, have maximum absorption at 515 nm in their first-order derivative spectrum. Theconcentrations of chromagen and K(2)S(2)O(8) were optimized (final concentration of PMZH and K2S2O8 were 0.166 mM and 0.11 mM, respectively) for better stability and sensitivity of the radical cation produced. Agood linear correlation was found between the percentage inhibition and the increasing amounts of standard antioxidants, with correlation coefficients ranging from 0.989 to 0.999. The newly developed assay was employed to evaluate the antioxidant capacity of Citrullus colocynthes L. and Artemisia absinthium extracts. The proposed assay involved a more stable radical cation and required only 1 h for preparation of a working solution in comparison to the 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical cation decolorizaion assay, which was reported to be less sensitive at low pH and almost 12-16 h were required for preparation of a working ABTS solution. Other assays employed to evaluate the antioxidant potential andradical-scavenging capacities of the extracts were the ferric-reducing antioxidant power, 2,2'-diphenyl-1-picrylhydrazyl radical scavenging, total phenolic contents assay, total flavonoid contents and metal-chelating activity assays, and the lipid peroxidation value in linoleic acid emulsion systems. The results indicate that boththe plants have potent free radical-scavenging activity and the ability to prevent lipid peroxidation and radical chain reactions.

Adulticidal Activity of Olea vera, Linum usitatissimum and Piper nigera against Anopheles stephensi and Aedes aegypti under Laboratory Conditions.

BACKGROUND: There are several plant extractions which are being used for mosquito control. The aim of this study was to evaluate the efficacy of Olea vera, Linum usitatissimum and Piper nigera against Anopheles stephensi and Aedes aegypti under laboratory conditions. METHODS: These tests were carried out using WHO recommended bioassay method for adult mosquitoes. RESULTS: The extracts from black pepper was more effective as adulticide with lowest LC(50) values (2.26% and 8.4%) against Aedes aegypti and Anopheles stephensi after 24 h of exposure while after 48h (1.56% and 5.11%) respectively. In terms of LC(90) value black pepper was best with (8.66% and 30.1%) against Ae. aegypti and An. stephensi after 24 h of exposure while after 48h (4.59% and 17.3%) respectively. In terms of LT(50) black pepper took 15 h to kill 50% tested population of Ae. aegypti while against An. stephensi it took more than 2 days. In terms of percentage mortality black pepper kill 84% of the population of Ae. aegypti and 44.75% of the An. stephensi population. CONCLUSION: Black pepper showed best results in term of LC(50), LC(90), LT(50) and percentage mortality against Ae. aegypti and An. stephensi. Our study suggested that the plant extracts have potential to kill adult mosquitoes, are environment friendly and can be used for the control of mosquitoes.

A comprehensive approach to identifying and authenticating botanical products.

Whether they are being taken as dietary supplements by the general public or being evaluated in a clinical study, the authenticity of botanical products is a matter of paramount concern. Botanical specimens and the dietary supplements derived from them can vary in quality and in chemical constituent profiles because of a number of factors. Subtle variations in botanical specimens are known to have profound effects on the quality, efficacy, and safety of botanical dietary supplements and can potentially alter the results of clinical studies that rely on these materials. A complete array of authentication and evaluation tools can be utilized to provide a well-rounded scientific approach to the authentication of botanical products. It is vital that the authenticity of botanical supplements be established using appropriate analysis tools regardless of whether the end products are being considered for evaluation in clinical studies or are being developed for the consumer market.

Fasting-induced intestinal damage is mediated by oxidative and inflammatory responses.

BACKGROUND: Green tea has been shown to repair fasting-induced mucosal damage in rat intestine. The aim of this study was to elucidate the underlying mechanism. METHODS: Five groups of rats were used. Group 1 had free access to chow diet and water, and those in group 2 were fasted for 3 days. Animals in group 3 were fasted for 3 days, then were allowed drinking water for a further 7 days. Groups 4 and 5 were fasted for 3 days, then given drinking water containing green tea or vitamin E respectively for 7 days. Blood was collected for estimation of total plasma antioxidants, and jejunal samples were used for immunohistochemical analysis of superoxide dismutase (SOD), catalase and glutathione peroxidase (GPx), and for estimation of myeloperoxidase (MPO) activity. RESULTS: Use of green tea was associated with a significant increase in total plasma antioxidants (P < 0.001), and mucosal SOD (P < 0.001), catalase (P = 0.006) and GPx (P = 0.017), but a significant decrease in MPO activity (P < 0.001). Vitamin E produced similar changes, but the effects were smaller. CONCLUSION: Green tea reverses the fasting-induced damage to the intestinal mucosa by its antioxidant and anti-inflammatory effect.