The natural abundance of L-canavanine, an active anticancer agent, in alfalfa, medicago sativa (L.)

L-Canavanine, a potentially toxic antimetabolite of L-arginine that is stored by many leguminous plants, has demonstrative antineoplastic activity against a number of animal-bearing carcinomas and cancer cell lines. This investigation evaluated the natural abundance of this anti-cancer compound in commercially available sprouts, and in ten varieties of the seed of alfalfa, Medicago Sativa (L.). Canavanine abundance in commercially grown sprouts varied according to the source; the young plant stored appreciable canavanine that ranged from 1.3 to 2.4% of the dry matter. Alfalfa seeds were also rich in this nonprotein amino acid as the canavanine content varied from 1.4 to 1.8% of the dry matter. On average, the tested seeds contained 1.54 ± 0.03% canavanine. Alfalfa seed canavanine content was comparable to the levels found in the seeds of representative members of the genus Canavalia , which are amongst the more abundance sources of this antimetabolite.     

Canavanine content in sword beans (Canavalia gladiata): analysis and effect of processing.

The amino acid canavanine is a potentially toxic constituent of leguminous seeds. The aim of the present study was to determine the ability of different processing methods to reduce canavanine in sword beans (Canavalia gladiata). For this purpose a method for the detection and quantification of canavanine was developed using reversed-phase high-performance liquid chromatography of the dabsylated derivatives. The recovery of canavanine using this method was 88-91%. Optimum extraction of canavanine from raw and processed beans was obtained by addition of hot water prior to overnight soaking. The results obtained with this method agree well with previously published values for raw seeds. The method is sensitive, specific and can successfully be applied to the detection of canavanine in legumes. Overnight soaking and boiling in excess water followed by decanting gave the most pronounced reduction in canavanine content (around 50%), followed by boiling and decanting excess water (34%). Roasting as used in this study and autoclaving were less effective in reducing the canavanine content.     

Effects of high levels of Brussels sprouts in the diet of rats

To examine the possible harmful effects of feeding Brussels sprouts to rats, groups of 10 male, weanling rats received the non-dehydrated vegetable in moist diets at levels providing 2.5–30% of the dry matter for 4 wk. A first study comprised test diets with 15 and 30% of the dry matter as uncooked or cooked Brussels sprouts and control diets without and with 0.2% potassium thiocyanate (KSCN) for comparison. The second study comprised diets with 0, 2.5, 5, 10 and 20% of the cooked vegetable and diets with 0 and 20% of the cooked vegetable with extra iodine. Diets with the uncooked vegetable contained considerably less intact glucosinolates than did diets with the cooked product, probably as a result of more extensive enzymatic degradation in the uncooked product. Growth depression and decreased food intake, not accompanied by decreased food efficiency, occurred in rats fed 10% or more dry matter as Brussels sprouts. These findings were less marked with the cooked than with the uncooked vegetable, probably because of unpalatability. Decreased levels of blood haemoglobin and plasma thyroxin were found with 15% or more Brussels sprouts. Prothrombin times were increased if 2.5% or more was fed. Thyroid stimulating hormone was increased by feeding potassium thiocyanate, but not by feeding the vegetable. Increased kidney weights and impaired kidney function not accompanied by microscopic renal changes were observed in rats fed 10% or more Brussels sprouts. Increased liver weights, which occurred from the 5% level, were accompanied by microscopic hepatic changes only at feeding levels from 10% of the cooked vegetable. ‘Morphological activation’ of the thyroid was increased with 10% or more of the cooked vegetable and with 0.2% KSCN. Iodine supplementation of the diets did not influence the results obtained with the vegetable. These studies indicated that 2.5% Brussels sprouts dry matter in the diet was not without effect, and that the thyroid characteristics were less sensitive to Brussels sprouts than were other criteria examined.     

The majority of in vitro macrophage activation exhibited by extracts of some immune enhancing botanicals is due to bacterial lipoproteins and lipopolysaccharides

We have identified potent monocyte/macrophage activating bacterial lipoproteins within commonly used immune enhancing botanicals such as Echinacea, American ginseng and alfalfa sprouts. These bacterial lipoproteins, along with lipopolysaccharides, were substantially more potent than other bacterially derived components when tested in in vitro monocyte/macrophage activation systems. In experiments using RAW 264.7 and mouse peritoneal macrophages the majority (85-98%) of the activity within extracts from eight immune enhancing botanicals was eradicated by treatment with agents (lipoprotein lipase and polymyxin B) known to target these two bacterial components. Alfalfa sprouts exhibited the highest activity of those botanicals tested but the appearance of this activity during the germination of surface sterilized seeds was abolished by the presence of antibiotics. These studies indicate that the majority of the in vitro macrophage activating properties in extracts from these botanicals can be attributed to the presence of lipoproteins and lipopolysaccharides derived from bacteria and that bacterial endophytes may be a significant source of these components.     

Germination rates of perilla (<Emphasis Type="Italic">Perilla frutescens</Emphasis> (L.) Britton) mericarps stored at 4°C for 1–20 years

Periodic update of stored seeds is indispensable for sustainable use and preservation of plant genetic resources, and performance of this task requires precise data on the relationship between storage period and the germination rate of seeds after long-term storage. In this study, germination rates of pure strains of perilla (Perilla frutescens (L.) Britton var. crispa) were investigated for three strains by using the mericarps stored for 1–20 years at 4°C under dry conditions. The germination rates were fairly good (65–70%) for 5–8 years after harvest, but dropped to almost 0% for mericarps stored more than 9 years. Mericarps that did not germinate were analyzed for viability by using 2,3,5-triphenyltetrazolium chloride. For old mericarps (stored more than 9 years), the germination rates and seed viability did not correlate. Further study on days required for germination of mericarps harvested each year was derived from our observation record, suggesting that germination rates and longevity of mericarp life were increasingly influenced by individual difference as the storage period lengthened.     

Seed borne nature of Azotobacter chroococcum in chilli (Capsicum annum) and its role in seed germination and plant growth

Investigations were carried out on seed borne nature of Azotobacter chroococcum in chilli and their role in plant growth at the Department of Agriculture Microbiology, University of Agricultural Sciences, Dharwad. Azotobacter chroococcum were isolated from both surface sterilized and unsterilized seeds of 14 varieties of chilli at different stages of fruit maturity. Inoculation with these strains increased the seed germination, root, shoot length and total dry matter content of chilli plants significantly in both sterilized and unsterilized soil., the results of which are presented.     

Influence of storage conditions on protein profile of pepper seeds,analyzed by SDS-PAGE

Seeds were stored under variable conditions for long or short periods of time at 5 degrees C or 25 degrees C after which they were germinated and total proteins were analyzed by SDS-PAGE. An 80 KDa protein was expressed only in the seeds that had been recently harvested (1 year storage period). A protein of 60 KDa was detected in much higher amounts in the seeds harvested in 1987 and stored at 5 degrees C for 14 years, as well as in seeds harvested in 2000 compared with the seeds harvested in 1989 and stored at 25 degrees C for 12 years. A 50 KDa protein was present in the seeds harvested in 1987 and stored at 5 degrees C, as well as in the seeds harvested in 2000 but not in the seeds harvested in 1989 and stored at 25 degrees C. It is proposed that SDS-PAGE can be used to identify proteins in germinating pepper seeds, that vary with the storage conditions. Such information could be of value for seed control and the evaluation of seed vigour.     

Seed-surface disinfection and germination effects of grapefruit seed extract (GSE) on <Emphasis Type="Italic">Lactuca sativa</Emphasis> seeds

It investigated that the seed sterilization and germination effects on Lactuca sativa seeds of two commercially available grapefruit seed extracts (GSEs) from the USA and Brazil. The total bacterial counts in the 0.05% GSE (USA) treatment groups were 0.0 CFU/g for all tested incubation times (1-6 h). All GSE treatments groups (from both the USA and Brazil) showed no or slightly harmful effects on L. sativa seed germination and seedling growth after 24 and 48 h. Additionally, seeds treated with 0.05% GSE showed slightly stronger anti-germination effects and seedling growth inhibition than those treated in a solution of the same concentration but with the pH adjusted to 7.0. We also evaluated the anti-germination effects of naringin, quercetin, and citric acid on L. sativa seeds to identify the active compounds in GSE. We observed no anti-germination effects in the naringin-treated group. In the quercetin-treated group, there was a slight dose-dependent anti-germination effect (0.01-0.1%). In groups treated with 0.01% and 0.05% citric acid (pH 4.79 and 4.03, respectively), no and slight anti-germination effects, respectively, were observed. However, there were strong anti-germination effects among seeds treated with 0.1% citric acid (pH 4.79). We found that 0.05% GSE (from both the USA and Brazil) had strong seed sterilization effects and no anti-germination effects. Together, these results indicate that GSE may be useful as a treatment for seed sterilization without inhibiting seed germination or seedling growth.     

The determination of morphine in urine and oral fluid following ingestion of poppy seeds

In workplace drug-testing programs, the use of heroin, morphine, and codeine is currently determined by the analysis of urine specimens. It has been shown that ingestion of poppy seeds can cause a positive test result for morphine. In an attempt to differentiate positive results caused by poppy seed ingestion from those caused by heroin or morphine abuse, the screening cutoff concentration for urine opiates in the federal workplace drug-testing program was raised to 2000 ng/mL from 300 ng/mL. Currently, oral fluid is under consideration as a possible alternative to urine for drug testing. The suggested cutoff for oral fluid morphine is 40 ng/mL; however, the effect of poppy seed ingestion on morphine concentrations in this specimen type has not been widely investigated. Volunteers at two separate sites ingested commercially available poppy seeds and/or poppy seed bagels. Oral fluid and urine samples were collected at both sites. Oral fluid samples were collected for 24 h; urine was collected for 2 days. The samples were analyzed for the presence of codeine and morphine using gas chromatography-mass spectrometry. Morphine concentrations greater than the suggested cutoff concentrations were detected in oral fluid up to 1 h and in urine for up to 8 h. This study has demonstrated that a positive result for morphine in oral fluid may be due to the ingestion of poppy seeds.     

Grape seed and skin extracts inhibit platelet function and release of reactive oxygen intermediates

Red wine and purple grape juice contain polymeric flavonoids with antioxidant properties believed to be protective against cardiovascular events but the alcohol and sugar content of these beverages has curtailed their medicinal use. Acute cardiac events are also associated with enhanced inflammation and thrombosis. In this study, the extracts from grape skins or seeds were examined for their anti-inflammatory properties and effect on platelet release of reactive oxygen intermediates. Incubation of platelets with seed or skin extract led to a decrease in platelet aggregation from 68.8+/-19.8% to 45+/-3.6% for seeds and to 27+/-7.2% for skin, respectively (P<0.05). Platelet incubation with grape skin or seed extracts led to a marked decrease in superoxide release from 73+/-6.2 to 2+/-3.4 for grape seeds and to 0.33+/-0.57 for grape skin (chemilum. units; P<0.05) as well as a significant increase in radical-scavenging activity, decrease in reactive oxygen species release by confocal microscopy, and enhanced platelet NO was measured using an NO-sensitive microelectrode. These effects were dose dependent for both grape extracts. Coincubation with seeds and skins led to additive inhibition of platelet aggregation, enhanced NO release, and prevented superoxide production. Incubation with seed or skin extracts led to an immediate attenuation of release of the inflammatory mediator, soluble CD40 ligand. Thus, the extracts from purple grape skins and seeds inhibit platelet function and platelet-dependent inflammatory responses at pharmacologically relevant concentrations. These findings suggest potentially beneficial platelet-dependent antithrombotic and anti-inflammatory properties of purple grape-derived flavonoids.     

普罗布考片剂溶出度及稳定性考察

利用紫外法和高效液相色谱法对普罗布考片剂的溶出度及稳定性进行考察，结果表明：溶出４５ｍｉｎ时，溶出度达７０％以上；高温８０℃放置１０天，含量下降２％，降解产物＞１％；高湿（ＲＨ９２５％）条件下溶出度略有下降，其它条件各项指标均未发生明显改变；加速实验及室温留样２年，其各项指标均未发生明显改变，可暂定有效期为２年．普罗布考片剂应于低温、密闭、干燥处保存．     

Toxicity and pharmacokinetics of the nonprotein amino acid L-canavanine in the rat

The toxicity of L-canavanine was investigated because of its demonstrated potential as an antitumor drug. This natural product was only slightly toxic to Sprague-Dawley rats following a single sc injection: the LD50 was 5.9 +/- 1 8 g/kg in adult rats and 5.0 +/- 1.0 g/kg in 10-day-old rats. Following a single dose of 2.0 g/kg, the systemic clearance value for canavanine in adult rats was 0.114 liter/hr, the volume of distribution at steady state was 0.154 liter, and the half-life was 1.56 hr. Forty-eight percent of the dose was excreted unaltered in the urine following an iv injection, and 16% of a sc dose was recovered in the urine. Bioavailability of a 2.0 g/kg sc dose was 72%. Single oral doses of canavanine were less toxic to adult rats than sc injections. Bioavailability of a 2.0 g/kg po dose was 43%, and only 1% of the administered canavanine was recovered in the urine. Twenty-one percent of the administered canavanine remained in the gastrointestinal tract 24 hr after an oral dose. Less than 1% of a 2.0 g/kg dose of L-[guanidinooxy-14C]canavanine was incorporated into the proteins of adult and neonatal rats 4 or 24 hr following administration. Repeated sc administration of canavanine resulted in more severe toxicity. Weight loss and alopecia were observed in rats given daily sc canavanine injections for 7 days. Food intake was decreased by 80% in adult rats subjected to this dosing regimen, but returned to normal after canavanine injections were terminated. Histological studies of tissues from adult rats treated with 3.0 g/kg canavanine daily for 6 days revealed pancreatic acinar cell atrophy and fibrosis. Serum amylase and lipase levels were elevated following one sc injection of 2.0 g/kg canavanine; after three daily injections both serum enzymes were depleted. Elevations in serum glucose and urea nitrogen, and depletion of cholesterol, were observed. The most significant changes were severe attenuations of serum aspartate aminotransferase, alanine aminotransferase, and alkaline phosphatase activity.     

Agronomic, qualitative (β-ODAP) and molecular variability in grasspea populations of the Marche region (central Italy)

Grasspea is an interesting crop for rational crop rotations in inner areas of the Marche region. However, breeding efforts are needed to improve its agronomic performance and to lower as much as possible the neuro-active amino acid β-ODAP in the seeds. Since the year 2000, a germplasm collection and evaluation program has been carried out. Accessions were evaluated by field trials for morphological, physiological and agronomic traits and for seed β-ODAP content. Significant variability was detected. In particular, populations collected in local farms showed a higher seed production compared with unadapted accessions. β-ODAP content varied between 0.26% and 0.53%. Moreover, AFLP molecular markers were applied to investigate genetic variation characterizing the populations included in the collection. In the year 2008 a breeding plan was started using two household populations, one with the lowest average β-ODAP content and one with the highest agronomic performance. Based on these results, a within population analysis for this trait will be started using the accession with the lowest β-ODAP content. Interesting information was also gathered using AFLP molecular markers, since a wide range of genetic variation supported the possibility of starting a mapping project aimed at the identification of genes of the biochemical pathway for β-ODAP.     

Chemopreventive effects of mustard (Brassica compestris) on chemically induced tumorigenesis in murine forestomach and uterine cervix

As there is a strong correlation between diet and cancer, the dietary constituents that inhibit mutagenesis and/or carcinogenesis are of paramount importance for the prevention of human cancer. In the present study, cancer chemopreventive potentials of different doses of mustard (Brassica compestris) seed mixed diets were evaluated against benzo(a)pyrene [B(a)P]-induced forestomach tumorigenesis and 3-methylcholantrene (MCA)-induced uterine cervix tumorigenesis. Results showed a significant inhibition of stomach tumour burden (tumours/ mouse) by mustard seeds. Tumour burden was 7.08 +/- 2.47 in the B(a)P-treated control group, whereas it was reduced to 1.36 +/- 1.12 (P<0.001) by the 2.5% dose and 1.18 +/- 0.87 (P<0.001) by the 5% dose of mustard seeds. The cervical carcinoma incidence, as compared to MCA-treated control group (73.33%), was reduced to nil (P<0.05) by the 5% diet of mustard seeds and to 13.33% (P<0.05) by the 7.5% diet of mustard seeds. The effect of the 2.5% and 5% mustard seed mixed diets was also examined on the antioxidant enzymes, glutathione content, lactate dehydrogenase (LDH) and lipid peroxidation in the liver of Swiss albino mice. The glutathione-S-transferase-specific activity was increased (P<0.05) by the 2.5% dose, whereas there was no significant change in the activity of DT-diaphorase. In antioxidant systems, significant elevation of the specific activities of superoxide dismutase and catalase was observed with both doses of mustard seeds (P<0.05). The level of reduced glutathione (GSH) measured as nonprotein sulphydryl content was elevated by the 2.5% dose of mustard seeds only (P<0.05). Lipid peroxidation measured as formation of thiobarbituric acid reactive substances production showed significant inhibition (P<0.05) by the 5% dose of mustard seed mixed diet. LDH activity was decreased significantly (P<0.05) by both the doses. The results strongly suggest the cancer chemopreventive potentials of mustard seeds and their ability to enhance the antioxidant defence system and in turn provide protection against the toxic effects of carcinogens. It is likely that the use of mustard seeds in the diet may contribute to reducing the risk of cancer incidence and burden in the human population.     

Variation of Aflatoxin Levels in Stored Edible Seed and Oil Samples and Risk Assessment in the Local Population

Five hundred and twenty samples of edible seeds and oilseeds (sunflower, palm, peanut, sesame, cotton, and grapeseed) were purchased from markets, farmers, and superstores in the central cities of Punjab, Pakistan. A total of 125 (48.1%) edible seed samples from a 6 ≤ months storage period, and 127 (48.8%) from a 2 ≥ years storage period were found to be infested with AFs. The average elevated amount of AFB₁ and total AFs was observed in a 2 ≥ years storage period, i.e., 28.6 ± 4.5 and 51.3 ± 10.4 µg/kg, respectively, in sesame seeds. The minimum amount of AFB₁ and total AFs was observed in palm seed samples with a storage period of 6 ≤ months, i.e., 9.96 ± 2.4, and 11.7 ± 1.90 µg/kg, respectively. The maximum amount of AFB₁ and total AFs were observed in peanut oil samples, i.e., 21.43 ± 2.60 and 25.96 ± 4.30 µg/kg, respectively, with a storage period of 2 ≥ years. Therefore, the maximum dietary intake of 59.60 ng/kg/day was observed in oil samples stored at a ≥ 2 years storage period. The results of the present study concluded that a significant difference was found in the amounts of total AFs in edible seed samples stored at 6 ≤ months and 2 ≥ years storage periods (p < 0.05).     

Glucosinolates in Moringa stenopetala

Bio-guided fractionation of seed extracts from Moringa stenopetala resulted in a myrosinase hydrolysis product, 5,5-dimethyloxazolidine-2-thione. It is formed from the glucosinolate glucoconringiin, which was identified together with O-(rhamnopyranosyloxy)benzyl glucosinolate from M. stenopetala for the first time. The glucosinolates in seeds, leaves and roots of M. stenopetala were quantified as des-sulphoglucosinolate by HPLC. The seeds without testa contained the highest concentration of glucoconringiin and of O-(rhamnopyranosyloxy)benzyl glucosinolate within the plant, 3 % and 19 % of dry mass, respectively. Abbreviations. GLS:glucosinolate GC:glucoconringiin RB-GLS: O-(rhamnopyranosyloxy)benzyl glucosinolate     

Natural occurrence of moulds and aflatoxin B1 in melon seeds from markets in Nigeria.

Shelled melon seeds (Colocynthis citrullus L.) were purchased from markets in randomly selected villages and towns in three states in each of the rain forest (Ogun, Oyo and Osun) and Northern guinea savanna (Kaduna, Niger and Bauchi) zones of Nigeria. The seed samples were analysed for incidence of visibly diseased seeds, moisture content, moulds and aflatoxin B1 contamination. The incidence of diseased seeds ranged from 6.4% to 50.4% in the forest, and 4.3% to 34.3% in the savanna, and the moisture content was 5.6% to 12.6% and 4.5% to 10.3%, respectively. Mould evaluation revealed that Aspergillus was the most frequent genus, followed by Penicillium, Botryodiplodia, Cladosporium and Rhizopus in decreasing sequential order. Aspergillus flavus had the highest individual count in melon seed from both zones. Aflatoxin B1 was detected at levels above 5 microg/kg in 32.2% of samples, while only 3.5% of the samples contained the toxin above the 20 microg/kg Nigerian tolerance level in food. The percentage of samples contaminated with aflatoxin B1 was statistically comparable for the pooled data of villages and towns. The median level of aflatoxin B1 was less than 5 microg/kg in the seed samples, while the mean aflatoxin B1 levels was 14.1 microg/kg in the forest and 13.0 microg/kg in the savanna samples.     

Stability and viscosity of a flavored omeprazole oral suspension for pediatric use.

PURPOSE: The stability and viscosity of preparations of a commercially available, flavored, immediate-release powder for oral suspension (omeprazole-sodium bicarbonate) during refrigerator and room temperature storage were investigated. METHODS: Omeprazole-sodium bicarbonate 20-mg packets were suspended to initial omeprazole concentrations of 0.6 and 2 mg/mL, and omeprazole-sodium bicarbonate 40-mg packets were suspended to initial omeprazole concentrations of 1.2, 2, 3, and 4 mg/mL. Suspensions were stored at 4 degrees C in darkness (refrigerated) or 22-25 degrees C (room temperature) in light for one week. A third set of suspensions was stored refrigerated for one month. Omeprazole's stability was quantified after 0, 6, 12, 24, 48, and 168 hours in one-week samples and after 0, 7, 14, 21, and 28 days in one-month samples using high-pressure liquid chromatography. Viscosities of refrigerated suspensions were measured after 0, 1, and 7 days. RESULTS: Refrigerated suspensions retained >98% and >96% of their initial omeprazole concentrations after one week and one month, respectively. Stability of room temperature suspensions was concentration dependent. After one week, the 0.6- and 1.2-mg/mL suspensions retained 87.2% and 93.1% of their respective initial omeprazole concentrations, whereas the 2-, 3-, and 4-mg/mL suspensions retained >97% of their initial omeprazole concentrations. Suspension viscosities varied 10-fold over the concentrations studied, but all were within the viscosity ranges of other commercially available oral suspensions. Prolonged refrigeration did not increase the suspensions' viscosities. CONCLUSION: Omeprazole-sodium bicarbonate suspensions of 0.6-4 mg/mL omeprazole were stored at 4 degrees C in darkness for up to 28 days. The viscosities of refrigerated suspensions did not increase over 7 days. Except for the 0.6 mg/mL preparations, suspensions stored at room temperature in the light retained >90% of their initial omeprazole content after 7 days, despite turning yellow.     

不同产地北细辛的种质特征

目的比较不同产地北细辛的种质特征。方法根据《中华人民共和国国家标准-农作物种子检验规程》和自建的马兜铃酸A检测方法,对8个产地的北细辛种子进行了水分、生活力、千粒质量、发芽率和马兜铃酸A含量的测定,并用SPSS13.0软件对检测结果进行聚类分析。结果各产地的种子总体上粒大饱满,活力较好,但水分都比较高,且都含有马兜铃酸A。结论清原县北三家乡和本溪谢家所产的北细辛种子从总体上来看,水分和马兜铃酸A的含量较低,生活力等其他指标较高,质量相对比较好。     

Ecotoxicological and Fertilizing Effects of Dewatered,Composted and Dry Sewage Sludge on Soil Mesofauna: A TME Experiment

The effects of dewatered, composted and dry urban sewage sludge on the soil mesofauna were tested in mesocosms. PVC containers were filled with soil/sludge mixtures in a proportion to amount to 6% organic matter content and were colonized with soil fauna coming from undisturbed forest soils. Mesocosms were incubated under laboratory conditions for 7, 30, 60, 120 and 180 days, after which fauna was extracted in Berlese funnels. The animals were classified at different taxonomic levels. Acari were classified to the suborder level for Astigmata, to the family level for Mesostigmata and Prostigmata and to the species level for Cryptostigmata. Acute- and medium-term effects were determined on the faunal density, relative abundance of the main taxa and community structure. Prostigmata were sensitive to the acute effect of the sludge, whereas Mesostigmata and particularly Cryptostigmata were sensitive to its medium-term effect. The most negative effects were found for dry sludge, which caused acute and medium-term effects on the invertebrate communities and on the soil trophic structure.