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Application of macronutrients in less amount along with improper application method(s) are the major factors for low yield. Experiments were designed with the objectives to determine the optimum rate of nitrogen (N) and potassium (K) along with suitable application method to minimize nutrients losses. The experimental site was New Developmental Farm of the University of Agriculture Peshawar and experiments were conducted for two consecutive years i.e. 2010-11 and 2011-12. After analysis, the results indicated that both N and K significantly affected days to phenology (anthesis and maturity). Days to phenology increased linearly with increase in rate of N and K and more days to phenology were recorded when both were at their highest rates. Similarly plant height, leaf area/tiller, flag leaf area, leaf area index, leaf area duration, grain yield, N uptake by grains and straw were increased with increasing level of N and K and attained maximum value when N and K was applied at the rate of 180 and 90 Kg/ha, respectively. Nutrients application method significantly affected plant height, leaf area/tiller, flag leaf area, grain yield and N uptake by straw and grains with maximum values for these traits were recorded when nutrients (N and K) were applied in such a combination that 15% nutrients were supplied through foliar and 85% through soil application. Application methods had a non-significant effect on days to phenology, leaf area index and leaf area duration. It is concluded from the results that application of N and K at the rate of 180 and 90 Kg/ha respectively in such a way that 15% is applied through foliar spray and 85% through soil application improved growth, yield and N uptake of wheat crop under the semi-arid climate.

Phosphorus (P) unavailability and lack of organic matter in calcareous soils in semiarid climates are the major reasons for low crop productivity. This field experiment was conducted at the Agronomy Research Farm of The University of Agriculture Peshawar, during the summer of 2015 to investigate the impact of plant residues (PR) (faba bean, garlic and paper mulberry residues) and phosphorous sources [(60 and 120kg ha-1) from single super phosphate (SSP) and poultry manure (PM)] with (+) and without (-) phosphate solubilizing bacteria (PSB) on the phenological development, growth and biomass yield of hybrid maize “CS-200”. Among the PR, application of faba bean residue was found to delay phenological development (days to tasseling, silking and physiological maturity), improved growth (taller plants, higher leaf area per plant and leaf area index) and produced the highest biomass yield (faba bean>garlic>paper mulberry residues). Application of P at the rate of 120kg ha-1 from the inorganic source (SSP) was more beneficial in terms of better growth and higher biomass yield (120-SSP≥120-PM>60-SSP>60-PM kg P ha-1). The plots with (+) PSB showed enhanced phenological development, produced significantly taller plants with higher leaf area per plant and leaf area index and produced the highest biomass yield. On the basis of these results we concluded that the application of faba bean residues, phosphorus at the rate of 120kg ha-1 either from organic or inorganic sources with the inoculation of seed with PSB improved the growth and total biomass of hybrid maize in the study area.

Variability in climate context is causing direct effects on reducing crops production. Possible strategies to counteract the impact of climate change is to configure the sowing time of crops and to seek out crop cultivars that are adapted to extreme climatic conditions of a specific ecological zone. Experiments were conducted at three ecological zones of Khyber Pakhtunkhwa Province of Pakistan i.e. (i. Agronomy Research Farm, The University Agriculture Peshawar ii. Agriculture Research Institute, Mingora, Swat iii. Farmers field, Karak, during the summer season, 2017–2018). Experiment was laid-out in Randomized Complete Block Design with split plot arrangement having four replications. Maize cultivars i.e. local open pollinated varieties (Azam, Jalal) and two hybrids (CS-200, CS-220) were sown on different dates starting from May 25 to July 25 with twenty days interval. Analysis of the data showed that early sowing (25th May) of maize in all zones (Karak, Peshawar and Swat) resulted in maximm emergence m−2, early emergence, highest leaf area, leaf area index, maximum light interception and higher chlorophyll content. However, delay in sowing time from 15th June onward in all zones resulted in lower emergence m−2, delay in emergence, reduced leaf area, leaf area index, lower light interception and chlorophyll content. Maize hybrids (CS-200 and CS-220) exhibited maximum emergence m−2, days to emergence, leaf area, chlorophyll content and light interception. At all zones (Karak, Peshawar and Swat) early sowing of maize hybrids (25th May to 15th June) is recommended for maintaining proper emergence parameters and improved leaf configuration for efficient resources utilization.

Under the rice&ndash, wheat cropping system (RWS), the continuous cropping of rice (Oryza sativa L.) and wheat (Triticum aestivum L.) deplete soil fertility, and reduce crop growth and total rice biomass. In RWS, both phosphorus (P) and zinc (Zn) deficiencies are considered important nutritional constraints for reducing rice crop growth rates (CGR) and total biomass/biological yield (BY). The objective of this experiment was to investigate the impact of phosphorus (0, 40, 80, 120 kg P ha&minus, 1) and zinc rates (0, 5, 10, 15 kg Zn ha&minus, 1) on CGR and BY of three rice genotypes [fine (Bamati-385) versus coarse (Fakhre-e-Malakand and Pukhraj)] in Northwestern Pakistan during summer 2011 (Y1) and 2012 (Y2). The results revealed that higher CGR at various growth stages and total BY was obtained with the integrated use of higher phosphorus (80 and 120 kg P ha&minus, 1) and zinc rates (10 and 15 kg Zn ha&minus, 1). The lower CGR and BY were recorded when P and Zn were not applied (control) or when P and Zn were applied alone. In the case of rice genotypes, the highest CGR and BY were recorded for the hybrid rice (Pukhraj) than the other two genotypes. The CGR was increased to the highest level at the heading stage as compared to tillering and physiological maturity. The increase in CGR had a positive impact on the total BY of rice cultivars. The increase in BY had a positive relationship with grain yield and grower&rsquo, s income. It was concluded from the study that the combined application of higher P and Zn rates to the coarse rice genotypes (Fakhre-e-Malakand and Pukhraj) could increase CGR, total BY, crop productivity and profitability.

Continuous cropping of rice (Oryza sativa L.) and wheat (Triticum aestivum L.) deplete soil fertility, reduced crop productivity, and grower&rsquo, s income. Phosphorus (P) and zinc (Zn) deficiencies are considered important nutritional constraints under rice-wheat cropping system. One strategy to increase crop productivity and grower&rsquo, s income under the rice-wheat system is the balanced application of P and Zn fertilizers. The objective of this research was to evaluate the impact of sole and various combinations of P (0, 40, 80, and 120 kg ha&minus, 1) and Zn levels (0, 5, 10, and 15 kg ha&minus, 1) on productivity (grain yield and yield components) and profitability (net returns) of different rice genotypes (cultivars) (fine (cv.Bamati-385) vs. coarse (cv. Fakhre-e-Malakand and cv. Pukhraj)) and their residual effect on the income of the succeeding wheat crop (cv. Siran-2010). The results revealed that both rice and wheat productivity and profitability was higher with the combined application of both nutrients at higher rates (80 and 120 kg P ha&minus, 1 and 10 and 15 kg Zn ha&minus, 1). The highest productivity and profitability was obtained with the cultivation of hybrid rice &ldquo, Pukhraj&rdquo, (Pukhraj &gt, Fakhre-e-Malakand &gt, Basmati-385). It was concluded from this study that application of higher P and Zn levels and growing of hybrid rice increased productivity and profitability under the rice-wheat cropping system.

Accumulation of heavy metals in soil and agricultural products is of great concern as it not only affects crop growth and yield but also food safety and marketability. The aim of the present study was to determine the effects of different organic amendments on toxic metal uptake by radish (Raphanus sativus L.). A pot experiment was conducted using garden soil as control and the contaminated soil collected from mine dumps of chromite mine, soap stone mine, quartz mine, manganese mine. Garden and mine impacted soil was mixed with vermicompost, leaf compost and spent mushroom compost at 1:1:1 ratio following a completely randomized block design. Results indicated that highest Cd (2.62 mg kg−1) and Cr (13.6 mg kg−1) concentration was reported in chromite mine soil, while Pb (23.3 mg kg−1) and Mn (59.2 mg kg−1) concentration as recorded in soap stone mine and manganese mine impacted soil, respectively. All the growth parameters and heavy metal concentrations were significantly affected by the amendments. The pollution load assessment of different mining soil revealed that the soil was moderately to considerably polluted with various heavy metals, particularly Cd with CF, Igeo and Eri values of 2.26, 0.45 and 67.68, respectively. Mining soils with high amount of heavy metals negatively affected radish growth and yield, while organic fertilizers amendment reduced heavy metals availability, increased radish growth and minimized human health risk. Among selected organic fertilizers vermicompost was more effective and reduced Cd, Cr, Pb and Mn uptake by 32.5, 50.25, 44.50 and 42.25 %, respectively, and improved radish growth, food quality and reduce human-health risk.

Pacific white shrimp (Litopenaeus vannamei) is an important cultural species worldwide. However, Vibrio spp. infections have caused a great economic loss in Pacific white shrimp culture industry. The immune responses of Pacific white shrimp to the Vibrio spp. is not fully characterized. In this study, the transcriptomic profiles of L. vannamei hemocytes were explored by injecting with or without Vibrio parahaemolyticus. Totally, 42,632 high-quality unigenes were obtained from RNAseq data. Comparative genome analysis showed 2258 differentially expressed genes (DEGs) following the Vibrio challenge, including 1017 up-regulated and 1241 down-regulated genes. Eight DEGs were randomly selected for further validation by quantitative real-time RT-PCR (qRT-PCR) and the results showed that are consistent with the RNA-seq data. Due to the lack of predictable adaptive immunity, shrimps rely on an innate immune system to defend themselves against invading microbes by recognizing and clearing them through humoral and cellular immune responses. Here we focused our studies on the humoral immunity, five genes (SR, MNK, CTL3, GILT, and ALFP) were selected from the transcriptomic data, which were significantly up-regulated by V. parahaemolyticus infection. These genes were widely expressed in six different tissues and were up-regulated by both Gram negative bacteria (V. parahaemolyticus) and Gram positive bacteria (Staphylococcus aureus). To further extend our studies, we knock-down those five genes by dsRNA in L. vannamei and analyzed the functions of specific genes against V. parahaemolyticus and S. aureus by bacterial clearance analysis. We found that the ability of L. vannamei was significantly reduced in bacterial clearance when treated with those specific dsRNA. These results indicate that those five genes play essential roles in antibacterial immunity and have its specific functions against different types of pathogens. The obtained data will shed a new light on the immunity of L. vannamei and pave a new way for fighting against the bacterial infection in Pacific white shrimp.

Hemocyanins (HMC): the copper-containing respiratory proteins present in invertebrate hemolymph, which plays many essential roles in the immune system. Currently, little is known about the HMC domains of Procambarus clarkii (P. clarkii) and their function in antimicrobial immune response. In this present study, we comparatively studied the expression pattern of native PcHMC with the three recombinant proteins of variable domains of crayfish hemocyanin (PcHMC-N, N-terminal domain of hemocyanin; PcHMC-T, tyrosinase domain of hemocyanin; PcHMC-C, C-terminal domain of hemocyanin). The results showed that three purified recombinant proteins had a strong binding to various bacteria and lipopolysaccharides that further highly agglutinated. The HMCs recombinant proteins showed strong antibacterial activity against V. parahaemolyticus and S. aureus by bacterial growth inhibition, phenoloxidase (PO) and phagocytosis assays. Specifically, rPcHMC1-T and rPcHMC1-C inhibited both the bacteria efficiently, rPcHMC1-T was highly upregulated the PO activity than the other recombinant proteins. Whereas, recombinant proteins pretreated crayfish hemocytes participated in phagocytosis activity, rPcHMC1-N and rPcHMC1-C proteins had a profound effect than the rPcHMC1-T on S. aureus and V. parahaemolyticus phagocytosis. The crayfish hemocyanin domains clearly exhibited antibacterial and phagocytic activities against both the bacteria, suggesting that its variable domains of hemocyanin have the different function on specific pathogen during the assault of pathogens.

Poor seed quality resulting in the establishment of poor stand which is the major problem of production in the cultivation of soybean. The objectives behind study were to evaluate the standard germination, field emergence and field emergence index of the seeds obtained from three land races (Kulat brown, Kulat white, Mothi) and two improved varieties (NARC-II, Swat-84) of soybean sown at four different dates at monthly interval from April to July during 2004, 2005 at New Developmental Farm, Agriculture University Peshawar. Standard germination, field emergence and field emergence index were significantly affected by sowing dates and varieties. In addition, to field emergence and field emergence index were observed to be affected by years. Sowing dates x varieties interactions were found significant for field emergence only. Maximum germination (74.6 %), field emergence (60.2 %) and field emergence index (79.1) were recorded for seeds harvested from May, April and April planted crops respectively. Kulat brown exhibited maximum germination (79 %), field emergence (66.1 %) and emergence index (83.6). In land races, germination, field emergence and field emergence index were detected as higher by 39, 58 and 12 percent comparatively to improved varieties. It is concluded that land races have the potential to emerge in various environmental conditions contrasted with improved varieties of soybean. Keywords: Germination; Field emergence; Field emergence index; Soybean land races http://dx.doi.org/10.19045/bspab.2017.60007

Laboratory experiments were conducted to examine the efficacy of fertilizer priming on seed germination behavior and vigor of two maize cultivars “Azam and Phari” at Agronomy Research Laboratory, University of Agriculture, Peshawar- Pakistan during 2015. Studied treatments include hydropriming (distilled water) and osmopriming with Calcium Ammonium Nitrate (CAN) at 0.5% for 24 hours. Non primed seed were used as control. Data were recorded on germination and vigor parameters according to standard procedures. Results showed that both hydro and osmo-priming significantly improved germination %, mean germination time, germination index, plumule and radical length, seedling fresh and dry weight of both cultivars as compared to control treatment. Cultivar Azam performed better in all germination and vigor attributes than Phari. It was concluded that both hydro and osmopriming with Calcium Ammonium Nitrate (CAN) at 0.5% for 24 hours are practical approaches for improving growth and vigor of maize crop under harsh environment.

From the leaves of Sarracenia purpurea, collected in Mistissini, Quebec, Canada, four goodyerosides and three phenolics and nine known compounds, were isolated. The structures of the compounds were determined by mass spectrometry, including HRMS, and by 1D and 2D NMR spectroscopy.

Planting date plays a significant role in soybean maturity and biomass yield. To study the maturity and biomass production potential under different environmental conditions, three indigenous land races (Kulat brown, Kulat white, Mothi) and two improved varieties (NARC-II, Swat-84) of soybean were planted at monthly from April to July during 2004 and 2005 growing season, at New Developmental Farm, The University of Agricultural Peshawar, Pakistan. Data was recorded for full maturity, plant height and biomass production. April planted crop took maximum days (187) to maturity, which decreased gradually with delay in planting. May planted crop gave maximum plant height (91.4 cm) and biomass yield (12598 kg ha -1 ) which decreased in later planting. Kulat white took maximum days (170) to maturity and attained maximum plant height (107 cm). However maximum biomass 12606 kg ha -1 yield was given by Mothi. Landraces took 41 days more than improved varieties to mature. In land races the plant height was almost double and the biomass yield was 31 percent higher than the improved varieties. Land races could prove higher yielders if the extra biomass production is directed towards seed

Purpose : The Cree of Eeyou Istchee in Northern Quebec identified Sarracenia purpurea L. as an important plant for the treatment of Type 2 diabetes. Traditionally the plant is used as a decoction (boiling water extract) of the leaf, however, in order to study the extract in a laboratory setting, an 80% ethanol extract was used. In this study, the phytochemistry of both extracts of the leaves was compared and quantified. Methods: Two S. purpurea leaf extracts were prepared, one a traditional hot water extract and the other an 80% ethanol extract. Using UPLC-ESI-MS, the extracts were phytochemically compared for 2 triterpenes, betulinic acid and ursolic acid, using one gradient method and for 10 additional substances, including the actives quercetin-3-O-galactoside and morroniside, using a different method. Results: The concentrations of the nine phenolic substances present, as well as an active principle, the iridoid glycoside morroniside, were very similar between the two extracts, with generally slightly higher concentrations of phenolics in the ethanol extract as expected. However, two triterpenes, betulinic acid and ursolic acid, were 107 and 93 times more concentrated, respectively, in the ethanol extract compared to the water extract. Conclusion: The main phytochemical markers and most importantly the antidiabetic active principles, quercetin-3-O-galactoside and morroniside, were present in similar amounts in the two extracts, which predicts similar bioactivity. This article is open to POST-PUBLICATION REVIEW . Registered readers (see “For Readers”) may comment by clicking on ABSTRACT on the issue’s contents page.

Purpose : The purpose of this study was to assess safety of the traditional antidiabetic extracts of either S. purpurea or its lead active principle, morroniside at the transcriptional level. The overarching objective was to profile and validate transcriptional changes in the cytochrome P450 family of genes, in response to treatment with S. purpurea ethanolic extract or its lead active, morroniside. Methods : Transcriptional activity was profiled using a 19K human cDNA microarray in C2BBe1 cells, clone of Caco-2 intestinal cells, which are a model of first-pass metabolism (1, 2). Cells were treated with S. purpurea extract for 4 and 24 hrs, as well as the pure compound morroniside for 4 hrs, to determine their effects. Results : No evidence of cytochrome P450 transcriptome regulation or of transcriptional activation of other diabetes relevant mRNA was detected after rigorous quantitative-PCR validation of microarray results. Conclusion : Our data do not support a transcriptional mechanism of action for either S. purpurea extract or its lead active, morroniside. This article is open to POST-PUBLICATION REVIEW . Registered readers (see “For Readers”) may comment by clicking on ABSTRACT on the issue’s contents page.

Herbario Juvenal Valerio Rodríguez Ethnopharmacological relevance The medicinal plant, Piper amalago L. (Piperaceae), is used traditionally by Q'eqchi' Maya healers for the treatment of "susto" a culture-bound syndrome. Previous research suggests that susto symptoms may be a manifestation of anxiety. The objectives were to characterize the effect of ethanolic extract of P. amalago in behavioral assays of anxiety at doses representative of traditional use and to isolate active principles. Materials and methods Rats treated orally with low dose ethanolic extracts of P. amalago leaves (8-75 mg/kg) were tested in several behavioral paradigms including the elevated plus maze (EPM), social interaction (SI), and conditioned emotional response (CER) tests, and compared to diazepam, a positive control. The active anxiolytic principle was isolated by bioassay guided isolation using an in vitro GABAA competitive binding assay. Results Extracts had significant anxiolytic activity in all behavioral tests, with the strongest activity in the SI and the CER paradigms. In an in vitro GABAA competitive binding assay, a 66.5 μg/mL concentration of P. amalago ethanol extract displaced 50% of the GABAA-BZD receptor ligand [3H]-Flunitrazepam. Bioassay-guided fractionation identified a furofuran lignan, a molecule with structural similarity to yangambin, with high affinity for the GABAA-BZD receptor as the principle bioactive. Conclusion The results suggest that the ethnobotanical use of this plant may have a pharmacological basis in its anxiolytic activity, as demonstrated in animal behaviour tests. Relevancia etnofarmacológica La planta medicinal, Piper amalago L. (Piperaceae), es utilizada tradicionalmente por los curanderos mayas Q'eqchi 'para el tratamiento del "susto", un síndrome ligado a la cultura. Investigaciones anteriores sugieren que los síntomas del susto pueden ser una manifestación de ansiedad. Los objetivos fueron caracterizar el efecto del extracto etanólico de P. amalago en ensayos conductuales de ansiedad a dosis representativas del uso tradicional y aislar principios activos. Materiales y métodos Se probaron ratas tratadas por vía oral con extractos etanólicos en dosis bajas de hojas de P. amalago (8-75 mg / kg) en varios paradigmas de comportamiento, incluido el laberinto en cruz elevado (EPM), la interacción social (SI) y la respuesta emocional condicionada ( CER) y en comparación con diazepam, un control positivo. El principio ansiolítico activo se aisló mediante aislamiento guiado por bioensayo utilizando un ensayo de unión competitiva de GABAA in vitro. Resultados Los extractos tuvieron una actividad ansiolítica significativa en todas las pruebas de comportamiento, con la actividad más fuerte en los paradigmas SI y CER. En un ensayo de unión competitiva de GABAA in vitro, una concentración de 66,5 μg / ml de extracto de etanol de P. amalago desplazó el 50% del ligando del receptor GABAA-BZD [3H] -Flunitrazepam. El fraccionamiento guiado por bioensayo identificó un lignano de furofurano, una molécula con similitud estructural a la yangambina, con alta afinidad por el receptor GABAA-BZD como el principal bioactivo. Conclusión Los resultados sugieren que el uso etnobotánico de esta planta puede tener una base farmacológica en su actividad ansiolítica, como se demuestra en pruebas de comportamiento animal. Relevância etnofarmacológica A planta medicinal, Piper amalago L. (Piperaceae), é tradicionalmente usada pelos curandeiros maias Q'eqchi 'para o tratamento de "susto", uma síndrome ligada à cultura. Pesquisas anteriores sugerem que os sintomas de susto podem ser uma manifestação de ansiedade. Os objetivos foram caracterizar o efeito do extrato etanólico de P. amalago em ensaios comportamentais de ansiedade em doses representativas do uso tradicional e isolar princípios ativos. Materiais e métodos Ratos tratados por via oral com extratos etanólicos de baixa dose de folhas de P. amalago (8-75 mg / kg) foram testados em vários paradigmas comportamentais, incluindo labirinto em cruz elevado (EPM), interação social (SI) e resposta emocional condicionada ( CER) e, em comparação com o diazepam, um controle positivo. O princípio ansiolítico ativo foi isolado por isolamento guiado por bioensaio usando um ensaio de ligação competitiva GABAA in vitro. Resultados Os extratos tiveram atividade ansiolítica significativa em todos os testes comportamentais, com a atividade mais forte nos paradigmas SI e CER. Em um ensaio de ligação competitiva de GABAA in vitro, uma concentração de 66,5 μg / mL de extrato de etanol de P. amalago deslocou 50% do ligante do receptor GABAA-BZD [3H] -Flunitrazepam. O fracionamento guiado por bioensaio identificou como principal bioativo um furofurano lignano, uma molécula com semelhança estrutural com a yangambina, com alta afinidade para o receptor GABAA-BZD. Conclusão Os resultados sugerem que o uso etnobotânico dessa planta pode ter embasamento farmacológico em sua atividade ansiolítica, conforme demonstrado em testes de comportamento animal. University of Ottawa, Canada Universidad Nacional, Costa Rica Belize Indigenous Training Institute, Belize Escuela de Ciencias Ambientales

Field experiment was conducted to investigate the impact of beneficial microorganism (BMO) and phosphorus (P) application on dry matter (DM) accumulation and partitioning in spring wheat (Triticum aestivum L., cv. Siren) under full irrigated (no moisture stress) and limited irrigated (dryland or moisture stress) conditions. The experiment was conducted at the Agronomy Research Farm, The University of Agriculture Peshawar during winter 2012-13. The experiment under both moisture conditions was laid out in randomized complete block design using three replications. The results revealed that DM accumulation and partitioning into leaf, stem and spike was significantly higher in wheat under irrigated than dryland wheat. The treated plots (rest) had higher total DM accumulation and partitioned more DM into leaf, stem and spike than control at both anthesis and physiological maturity (PM). Application of P and BMO at the highest rates (90 kg P ha-1 and 30 L ha-1, respectively) had accumulated more total DM and had partitioned more DM into leaf, stem and spike at the two growth stages. We found that under irrigated condition, increase in both P and BMO levels (90 kg P ha-1 and 30 L ha-1, respectively) and under dryland condition the intermediate levels of both P and BMO (60 kg P ha-1 and 20 L ha-1, respectively) had produced higher total DM and partitioned more DM into various parts at both anthesis and PM. The percent DM partitioning into leaf was more (36%) than stem and spike (each 32%) at anthesis stage; while at PM, more DM was partitioned into spike (59%) than stem (21%) and leaf (20%). Increase in DM partitioning into spike under both irrigated and dryland wheat with proper P and BMO management showed positive relationship with grain yield that resulted in higher growers income in the study area.

A single-laboratory-validated NMR spectroscopy method was established for determining the quantity of chlorogenic acid and hyperoside from crude extract material of blueberry leaves of the species Vaccinium angustifolium var. laevifolium House. The calibration curve of chlorogenic acid showed a highly linear regression, R = 0.99998. NMR spectroscopy identification and quantification of the constituents directly from the mixture, within the error of HPLC-diode array detector analysis, were determined as 7.53 mM chlorogenic acid (64.0 mg chlorogenic acid/g powdered leaf) and 0.77 mM hyperoside (8.58 mg hyperoside/g powdered leaf). The LOD was calculated to be 0.01 mM and the LOQ 0.01 mM by the 9 min and 13 s NMR spectroscopy experiment utilized. The assay showed no significant interference from different field strengths, extraction mesh size, gravimetric scale precision, NMR spectroscopy tube type, pulse program, amount of starting dry material, or day-to-day operation. The robustness of NMR spectroscopy as a means of definitively monitoring chlorogenic acid and hyperoside content directly from crude extracts was demonstrated by Youden statistical analysis.

Through ethnobotanical surveys, the CIHR Team in Aboriginal Antidiabetic Medicines identified 17 boreal forest plants stemming from the pharmacopeia of the Cree First Nations of Eeyou Istchee (James Bay region of Northern Quebec) that were used traditionally against diabetes symptoms. The leaves of Sarracenia purpurea (pitcher plant), one of the identified Cree plants, exhibited marked antidiabetic activity in vitro by stimulating glucose uptake in C2C12 mouse muscle cells and by reducing glucose production in H4IIE rat liver cells. Fractionation guided by glucose uptake in C2C12 cells resulted in the isolation of 11 compounds from this plant extract, including a new phenolic glycoside, flavonoid glycosides, and iridoids. Compounds 6 (isorhamnetin-3-O-glucoside), 8 [kaempferol-3-O-(6″-caffeoylglucoside], and 11 (quercetin-3-O-galactoside) potentiated glucose uptake in vitro, which suggests they represent active principles of S. purpurea (EC(50) values of 18.5, 13.8, and 60.5 μM, respectively). This is the first report of potentiation of glucose uptake by compounds 6 and 8, while compound 11 (isolated from Vaccinium vitis) was previously shown to enhance glucose uptake. Treatment of H4IIE liver cells with the new compound 1, 6'-O-caffeoylgoodyeroside, decreased hepatic glucose production by reducing glucose-6-phosphatase enzymatic activity (IC(50) = 13.6 μM), which would contribute to lowering glycemia and to the antidiabetic potential of S. purpurea.

Bioassay-guided fractionation of a crude extract (80% EtOH in H(2)O) of stem bark of Sorbus decora led the isolation of three new pentacycle triterpenes (compounds 1-3). The structures of 1-3 were established on the basis of spectroscopic methods (IR, HREIMS, 1D and 2D NMR) as 23,28-dihydroxyursan-12-ene-3β-caffeate, 23,28-dihydroxylupan-20(29)-ene-3β-caffeate, and 3β,23,28-trihydroxy-12-ursene, respectively. Compound 2 significantly enhanced glucose uptake in C2C12 cells, but compounds 1 and 3 did not. In addition, triterpenoids 4-8, catechin, and epicatechin were also isolated. This is the first comprehensive report of the phytochemical constituents of S. decora since the initial study by Narashmachari and von Rudloff (1962) and includes evaluation of their antidiabetic activity.

Among modern day metabolic diseases, obesity has reached epidemic proportions worldwide and novel therapeutic support strategies are urgently needed. Adipocytes are interesting targets in this context. Using ethnobotanical and bioassay screening techniques, we have identified two Boreal Forest plants used by the James Bay Cree that potently inhibit adipogenesis, namely ALNUS INCANA ssp. RUGOSA (Speckled Alder) and POPULUS BALSAMIFERA (Balsam Poplar). The mode of action of this inhibitory activity was reported in a companion paper. The current study report the results of a classical bioassay-guided fractionation approach aimed at identifying the active principles responsible for the inhibition of adipogenesis, as measured using triglyceride accumulation in the 3T3-L1 adipocyte model cell line. The glycosides oregonin and salicortin were isolated and identified as the respective active principles for ALNUS INCANA and POPULUS BALSAMIFERA. These compounds thus offer promise as novel agents to mitigate the incidence or the progression of obesity.

Several medicinal plants that stimulate glucose uptake in skeletal muscle cells were identified from among species used by the Cree of Eeyou Istchee of northern Quebec to treat symptoms of diabetes. This study aimed to elucidate the mechanism of action of one of these products, the berries of Vaccinium vitis idaea, as well as to isolate and identify its active constituents using a classical bioassay-guided fractionation approach. Western immunoblot analysis in C2C12 muscle cells revealed that the ethanol extract of the berries stimulated the insulin-independent AMP-activated protein kinase (AMPK) pathway. The extract mildly inhibited ADP-stimulated oxygen consumption in isolated mitochondria, an effect consistent with metabolic stress and the ensuing stimulation of AMPK. This mechanism is highly analogous to that of Metformin. Fractionation guided by glucose uptake activity resulted in the isolation of ten compounds. The two most active, quercetin-3-O-glycosides, enhanced glucose uptake by 38-59% (50 muM; 18 h treatment) in the absence of insulin. Quercetin aglycone, a minor constituent, stimulated uptake by 37%. The quercetin glycosides and the aglycone stimulated the AMPK pathway at concentrations of 25-100 muM, but only the aglycone inhibited ATP synthase in isolated mitochondria (by 34 and 79% at 25 and 100 muM, respectively). This discrepancy suggests that the activity of the glycosides may require hydrolysis to the aglycone form. These findings indicate that quercetin and quercetin 3-O-glycosides are responsible for the antidiabetic activity of V. vitis crude berry extract mediated by AMPK. These common plant products may thus have potential applications for the prevention and treatment of insulin resistance and other metabolic diseases.

Caffeic acid phenethyl ester (CAPE) has recently been shown to potently stimulate glucose uptake in cultured skeletal muscle cells through the AMPK pathway and therefore to have anti-diabetic potential. We report here that CAPE increases glucose uptake in C2C12 muscle cells by 225+/-21% at 50 microM, and that activation of AMPK is a consequence of the metabolic stress resulting from an uncoupling-type disruption of mitochondrial function (complete uncoupling at 50 microM). We also observe that the therapeutic potential of CAPE is offset by its high potential for toxicity. The purpose of this study was therefore to identify other active caffeic acid derivatives, evaluate their ratio of activity to toxicity, and elucidate their structure-activity relationship. Twenty naturally occurring derivatives were tested for glucose-uptake stimulating activity in C2C12 cells following 18 h of treatment and for uncoupling activity in isolated rat liver mitochondria. Cytotoxicity was assessed in C2C12 cells by the release of lactate dehydrogenase over 18 h. In addition to CAPE, four compounds were identified to be active, both stimulating glucose uptake and uncoupling isolated mitochondria. Activity required that the caffeic acid moiety be intact and that the compound not contain a strongly ionized group. Both activity and toxicity were found to be well-correlated to predicted lipophilicity. However, two compounds exhibited little to no toxicity while still stimulating glucose uptake by 65-72%. These results support a therapeutic potential for this family of compounds and provide the framework for the design of alternatives to Metformin with an optimized balance of safety and activity.

A novel pharmacological mechanism of action for the anxiolytic botanical Melissa officinalis L. (lemon balm) is reported. The methanol extract was identified as a potent in vitro inhibitor of rat brain GABA transaminase (GABA-T), an enzyme target in the therapy of anxiety, epilepsy and related neurological disorders. Bioassay-guided fractionation led to the identification and isolation of rosmarinic acid (RA) and the triterpenoids, ursolic acid (UA) and oleanolic acid (OA) as active principles. Phytochemical characterization of the crude extract determined RA as the major compound responsible for activity (40% inhibition at 100 µg/mL) since it represented approximately 1.5% of the dry mass of the leaves. Synergistic effects may also play a role. Copyright © 2009 John Wiley & Sons, Ltd.