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Examples of poisonous plants in Israel of importance to animals and man.

Several plants causing poisoning in herbivores in Israel have an association with human health. Ferula communis (giant fennel) contains a potent anticoagulant and induces severe haemorrhagic diathesis in grazing ewes, or similar changes in utero in lambs. These and other teratogenic changes have been seen in babies from mothers receiving the anticoagulant warfarin during pregnancy. The Ferula interaction in sheep may be used as a model to study this embryotoxicity in man. Ammi majus (bishops weed) causes severe ophthalmic changes, in particular pigmentary retinopathy in photosensitized domestic fowl. The photoactive principle is currently used in the treatment of psoriasis in man, and such patients should be examined for possible ocular toxicity. Trigonella foenum-graecum (fenugreek) caused myopathy in ruminants and is ingested by certain ethnic groups. Oxalis pes-caprae (Bermuda buttercup) causes large-scale oxalate poisoning in sheep and is avidly eaten by children.

Animals↗

Physiological and cytochemical studies on activator calcium in contraction by smooth muscle of a sea cucumber, Isostichopus badionotus.

The physiological properties of mechanical responses and the intracellular localization and translocation of calcium as a pyroantimonate precipitate were studied in the longitudinal retractor muscle (LRM) of a Bermuda sea cucumber. Acetylcholine (ACh)-induced contraction was reduced by lowering the external Ca concentration, and suppressed completely by prolonged soaking in Ca-free solution. The magnitude of ACh-induced contraction was decreased by Mn and La ions. Furthermore, procaine reduced the ACh-induced contraction. The complete removal of Ca and Mg ions from the external medium induced a so called Ca . Mg-removal contraction. Electron microscopically, numerous subsarcolemmal vesicles were observed in the LRM fibers. In the resting fibers, pyroantimonate precipitates were localized in the subsarcolemmal vesicles and along the inner surface of plasma membrane. While, in the fiber fixed during mechanical activity, the pyroantimonate precipitates were decreased remarkably in the subsarcolemmal vesicles and at the plasma membrane, and diffusely distributed in the myoplasm. Electron-probe X-ray microanalysis showed that the precipitate contains Ca in a significant amount. These results indicate that the contraction of the LRM fibers is caused not only by Ca-influx but also by Ca-release from the intracellular storage sites, such as the subsarcolemmal vesicles and the inner surface of plasma membrane.

Animals↗

Cytoplasmic localization of transcripts of a complex G+C-rich crab satellite DNA.

The primary sequence and higher order structures of a G+C-rich satellite DNA of the Bermuda land crab Gecarcinus lateralis have been described previously. The repeat unit of the satellite is approximately 2.1 kb. In exploring a possible function for this satellite, we asked whether it is transcribed. As a probe for transcripts, we used a segment of DNA amplified from a 368 bp EcoRI fragment from the very highly conserved 3' end of the satellite DNA. During polymerase chain reaction (PCR) amplification, the probe was simultaneously either radiolabeled or biotinylated. Tissue- and stage-specific transcripts were observed when blots of poly(A)+ mRNAs recovered from polysomes isolated from crab tissues [including midgut gland (hepatopancreas), limb bud, and claw muscle] were probed with the satellite DNA fragment. The presence of satellite transcripts in polysomal mRNAs is strong evidence that the transcripts had reached the cytoplasm. To corroborate the presence of transcripts in the cytoplasm, we investigated in situ hybridization of satellite probes with RNAs in tissue sections. Biotinylated satellite DNA probes were applied to sections of midgut gland, limb bud papilla, ovary, or testis of anecdysial crabs. Retention of RNAs in tissue sections was improved by UV-irradiation prior to hybridization. Transcripts were abundant in the cytoplasm of all tissues except testis. Sections of crab midgut gland treated with RNase A prior to hybridization and sections of mouse pancreatic tumor served as controls; neither showed any signals with the probe.

Animals↗

Characterization of a tandemly repeated DNA from the fleshfly Sarcophaga bullata.

In studies on the highly repetitive DNA sequences of the flesh fly Sarcophaga bullata, a 279 bp tandem repeat was cloned and sequenced. A 17 bp stretch within the clone was identical to a motif repeated five times in the satellite DNA of the Bermuda land crab. Southern DNA blotting showed the tandem repeat had a high degree of conservation of MboI sites, but had divergence for EcoRI sites; thus, all repeat units were not identical. The cloned DNA localized to the quinacrine-bright centromeric heterochromatin of the C and E autosomes and to sites on the chromosomal arms. In cases of asynapsis of homologs, the probe localized to euchromatic sites on both homologs or sometimes only on one homolog. The probe also localized near, to, or at a major developmental puff (B9). We conclude that blocks of this short interspersed repetitive DNA occur throughout the Sarcophaga genome in both heterochromatin and euchromatin, and also that the variable position of these sequences suggests they possess a degree of instability.

Animals↗

Mineral status of grazing beef cattle in the warm climate region of Florida.

An experiment was conducted to determine the mineral status of purebred Brahman beef cattle grazing Bermuda--Bahia pastures grown on sandy, well-drained soils in Central Florida. Soil, plant, blood serum and liver tissue concentrations of calcium, potassium, sodium, magnesium, manganese and cobalt were generally adequate. Sixty-nine per cent of forages and 40% of serum samples were deficient in phosphorus. Seventy-five per cent of soils, 38% of forages and 18% of liver samples were deficient in copper. Sixty-nine per cent of forage samples were low, less than 30 ppm, in iron. All samples of forage, liver and serum were deficient in selenium. Likewise, all soil and forage samples and 47 and 88% of serum and liver samples, respectively, were deficient in zinc. Mineral elements most likely deficient and needed in supplements for grazing cattle on sandy, well-drained soils in the warm climate region of Central Florida are phosphorus, copper, selenium and zinc.

Animals↗

Assay for trans-p-coumaroyl esterase using a specific substrate from plant cell walls.

Cell walls of Coastal Bermuda grass (Cynodon dactylon) were treated with polysaccharide hydrolases to release O-[5-O-(trans-p-coumaroyl)-alpha-L-arabinofuranosyl]-(1----3)-O-be ta-D- xylopyranosyl-(1----4)-D-xylopyranose (PAXX) which was isolated by liquid chromatography. The isolated PAXX was greater than 95% pure as determined by 1H NMR and was used as substrate for a sensitive assay of trans-p-coumaroyl esterase. PAXX was hydrolyzed by culture filtrates from the anaerobic fungus Neocallimastix MC-2. The trans-p-coumaric acid released by enzymatic hydrolysis was assayed by reverse-phase HPLC, and as little as 100 ng of acid could be determined. Steady-state velocities for the release of the acid obeyed Michaelis-Menten kinetics. Vmax was determined to be 1.17 mumol min-1 mg-1 and Km 13.2 microM at pH 7.5 and 30 degrees C.

Carbohydrate Sequence↗

Characterization of grass pollen allergens by affinity chromatography on Concanavalin A-Sepharose.

Glycoprotein allergens of different carbohydrate composition can be separated by lectin interaction in affinity chromatography. The present study shows that several differences exist in the sugar moiety both within the species and between different grass species. Experiments with Bermuda grass and common reed indicate that these 2 extracts possess allergens with a high affinity for Con A-Sepharose. This offers a simple and rapid method for isolation and purification of the major allergenic components of these 2 species in combination with more conventional separation techniques. The purity of single allergens may vary markedly depending on the occurrence of other interfering, non-allergenic substances having free terminal alpha-D-mannopyranoside or alpha-D-glycopyranoside residues which bind to the immobilized lectin.

Allergens↗

The cross-reactivity of IgE antibodies with pollen allergens. I. Analyses of various species of grass pollens.

Atopic patients with histories of grass pollen allergy often are sensitive to a variety of species of grasses. Using a serum pool from patients sensitive to June grass, we analyzed the reactivity of IgE antibodies to seven grasses by the radioallergosorbent test. Extracts were analyzed for their inhibitory activities with solid-phase allergens prepared from all of the grass pollen. Also samples of serum were exhaustively absorved with solid-phase allergens and the supernatants tested to determine the reactivity of the remaining IgE antibodies. Three patterns of reactivity were observed: (1) June, orchard, meadow fescue, and perennial rye grasses displayed similar reactivity in both inhibition and absorption studies; (2) sweet vernal and Bermuda grasses were considerably less reactive with the serum pool, indicating that they lacked antigenic determinants possessed by the other grasses; and (3) timothy grass possessed unique antigenic determinants. Knowledge of these patterns of cross-allergenicity is of importance for diagnosis and treatment of sensitive patients as well as for in vitro standardization of extracts.

Absorption↗

Pulmonary response to nasal-challenge testing of atopic subjects with stable asthma.

Nasal deposition of allergen or histamine could cause bronchoconstriction in subjects with asthma by a reflex mechanism. To investigate this possibility, six atopic subjects with stable asthma and five normal control subjects were studied by nasal-challenge testing with saline, Bermuda grass-pollen allergen, and histamine on paper disks. Challenges were done on 3 separate days by use of a double-blind, randomized trial design. Fivefold serial dilutions were used to determine threshold doses for provocation of at least a twofold increase in the postsaline nasal airway resistance. No patient developed cough or wheezing or required treatment at the end of the nasal-challenge tests, even when persistent sneezing was provoked or in the subject who had a dual nasal response to nasal challenge. In spite of changes in nasal airway resistance that generally conformed to expectations, there were no significant effects of nasal challenge on lung function, including forced expiratory flow rates, vital capacity, residual volume, and partial expiratory flow-volume curves.

Adult↗

A double-blind placebo-controlled trial of polymerized whole grass administered in an accelerated dosage schedule for immunotherapy of grass pollinosis.

Forty-four patients were entered into a study of the efficacy and safety of individually polymerized grass (IPG) immunotherapy with an accelerated dosage schedule. Patients were paired on the basis of cutaneous end point titrations to timothy, orchard, and Bermuda grass-pollen extracts. In a double-blind manner, one patient in each pair was treated in nine weekly visits with 13 injections that totaled 24,000 PNU of each grass to which the patient had cutaneous reactivity. The other patient in each pair received caramelized glucose histamine placebo. Symptom and medication score sheets were completed by 33 patients each day of the grass season. Blocking antibody rose significantly in the IPG-treated group but was unchanged in the placebo-treated group. By Wilcoxon paired signed-rank test, the symptom medication scores in the IPG-treated group were significantly lower than those in the placebo-treated group. There were no systemic reactions and no clinically significant changes in routine laboratory tests in either group. In summation, this study demonstrates the safety, immunogenicity, and efficacy of IPG therapy in an accelerated dosage schedule for treatment of grass pollinosis.

Adult↗

The prevalence of allergic skin test reactivity to eight common aeroallergens in the U.S. population: results from the second National Health and Nutrition Examination Survey.

Immediate hypersensitivity skin tests to eight select allergens were performed on a sample (N = 16,204) of the civilian noninstitutional population of the United States, 6 to 74 years of age, in the second National Health and Nutrition Examination Survey (NHANES II). The eight allergens were house dust, cat, dog, Alternaria, mixed giant/short ragweed, oak, perennial ryegrass, and Bermuda grass. Skin test reactivity was defined as a mean erythema diameter greater than or equal to 10.5 mm at the 20-minute reading. Overall, 20.2% of the participants reacted to at least one allergen. Peak reactivity occurred in the 12 to 24-year-old age group. Reactivity was higher in blacks versus whites, but the difference did not reach statistical significance (23.2% versus 19.8%; p greater than 0.05). Male participants had an increased prevalence of reactivity versus female participants in whites (22.0% versus 17.6%), but not in blacks (23.2% versus 23.3%). Skin test reactivity increased in both whites and blacks with increasing income and education. The prevalence of skin test reactivity was higher in urban versus rural areas, but the difference was statistically significant only for whites (whites, 21.6% versus 16.4%; blacks, 23.8% versus 18.4%; p greater than 0.05). With logistic regression, the most important predictors of skin test reactivity in whites were age, sex, urban residence, and poverty status. In blacks, the most important predictors were age, urban residence, and poverty status.

Adolescent↗

Use of standardized and conventional allergen extracts in prick skin testing.

This study examined whether commercially available conventional and standardized allergen extracts differ enough in potency to affect routine prick skin test results. Extracts of white oak, timothy, Bermuda, Russian thistle, short ragweed, sagebrush, Alternaria, and cat dander were examined in allergic patients and in nonatopic subjects with no personal or family history of asthma, rhinitis, or eczema. Conventional nonstandardized extracts (1:10 or 1:20 wt/vol) from two sources were compared with three concentrations (100,000, 10,000, and 1000 AU/ml) of a single standardized extract. Preparations were compared in the allergic patients with computerized planimetry, and in all patients and subjects with a conventional skin test grading system. Skin test area for the conventional extracts generally fell between the 10,000 and 100,000 AU/ml concentrations of the standardized extract. Skin test reactivity to at least one allergen extract occurred in 31% of the nonatopic subjects; there was no difference between the number of 3+ and 4+ reactions for conventional and standardized extracts. Results indicate that standardized and conventional extracts are frequently similar, but are not directly interchangeable.

Adolescent↗

Monoclonal antibodies to the major Lolium perenne (rye grass) pollen allergen Lol p I (Rye I).

Thirteen monoclonal antibodies (MAbs) were produced against Lol p I (Rye I), the major Lolium perenne (rye grass) pollen allergen. Spleen cells from A/J and SJL mice immunized with highly purified Lol p I (Lol I) were allowed to fuse with cells from the non-secreting Sp2/0-Ag14 myeloma cell line. Each MAb was analyzed for antigenic specificity by radioimmunoassay (RIA) using 125I-Lol I. The epitope specificities of seven of the MAbs were examined by competitive binding against a labelled standard MAb for the Lol I antigen (Ag). The dissociation constant, Kd, of one MAb (No. 3.2) that was studied most extensively was determined by double Ab RIA to be 3.5 X 10(-6) L/M. This MAb recognized the related 27,000-30,000 Group I glycoproteins found in the pollens of nine other species of grass pollens tested, including weak binding to Bermuda grass Group I (Cyn d I), which by conventional analysis using polyclonal anti-Lol I serum shows no detectable binding. Monoclonal antibody No. 3.2 was coupled covalently to Sepharose 4B and used to prepare highly purified Lol I from a partially purified rye pollen extract. Finally, an RIA was developed which permitted the analysis of the Group I components in rye grass and nine other grass pollen species. The latter assay is likely to prove useful in the standardization of grass pollen extracts according to their Group I contents.

Allergens↗

Olive (Olea europea) and privet (Ligustrum vulgare) pollen allergens. Identification and cross-reactivity with grass pollen proteins.

Protein blotting studies showed that three olive pollen components with mol. wts approximately 18-19, 20 and 40 kD can be considered to be major allergens. For privet pollen, the highest recognition frequencies were for allergens of mol. wts approximately 20, approximately 19, approximately 40 and approximately 70 kD. When results with the 62 subjects examined were separated into groups corresponding to their geographical locations, viz. Italy, France and Australia, subjects sensitized to olive, but not other pollens (some Italian subjects), were found to show higher frequencies of recognition of major olive allergens than subjects sensitized to olive pollen via cross-reacting allergens from unrelated pollen sources (the Australian and French subjects). Blotting, adsorption and elution and inhibition studies clearly demonstrated allergenic cross-reactivity (that is, antigenic cross-reactivity detected by IgE antibodies) between olive, privet, ryegrass (Lolium perenne) and couch grass (Bermuda grass: Cynodon dactylon) pollen components. As with our previous findings with birch pollen, we conclude that the presence of pollen-reactive IgE antibodies may not necessarily be a true reflection of the sensitizing pollen species.

Allergens↗

Effect of fenbendazole molasses supplement block treatment on nematode infection and subsequent weight gain of weanling beef calves.

Two trials were conducted to evaluate the effect of fenbendazole molasses supplement block treatment on weight gain of weanling beef calves. An initial (November) oral fenbendazole (5 mg kg-1) treatment was administered to all animals in both trials. In Trials 1 and 2, respectively, two fenbendazole molasses supplement block treatments were administered 6 (6.4 mg kg-1) and 16 (7.0 mg kg-1) weeks later and 7 (3.4 mg kg-1) and 14 (3.8 mg kg-1) weeks later. During the treatment periods, calves were maintained on dormant bermuda grass pasture, supplemented with hay and protein/mineral blocks. The first block treatment was administered approximately midway through the supplement period to suppress the infection level when nutrition was marginal. The second treatment was administered just prior to ryegrass grazing. Trial 1 and 2 treatments resulted in reducing the infection level, as indicated by reduced mean fecal egg counts. Treatment group mean fecal egg counts remained lower than control group mean counts throughout each trial. In Trial 1, the treatment group gained 29.5 kg more than the control group. In Trial 2, the control group gained 15.5 kg more than the treatment group. Fecal egg count data indicated that the infection level during Trial 1 was higher than during Trial 2. This difference could influence weight gain.

Administration, Oral↗

Demonstration of remarkable sequence divergence in variants of a complex satellite DNA by molecular cloning.

Repeat units of a complex G + C-rich satellite of the Bermuda land crab have been cloned by insertion into either the PstI or EcoRI site of pBR322 or the EcoRI site of pUC9. While most of the recombinants contained inserts of approx. 2.1 kb, the average size of repeat units seen in cellular satellite digests, several inserts were markedly different in size. Two domains that account for major sequence differences among the satellite variants and that may be 'hotspots' for sequence modification have been subcloned to permit characterization of their secondary and tertiary structures independent of the influence of the other unusual sequences present. One of these domains is striking in its content of simple repeats; one strand is highly biased in pyrimidines which may permit the formation of unusual secondary and/or tertiary conformations. The other subcloned domain is rich in Pu/Py; preliminary data indicate a transition from B----Z DNA in this region.

Animals↗

Denaturants or cosolvents improve the specificity of PCR amplification of a G + C-rich DNA using genetically engineered DNA polymerases.

We describe conditions that improve the specificity of amplification of a G + C-rich (57% G + C) DNA by PCR. Under standard conditions a 368-bp segment of the approx. 2.1-kb repeat unit of a satellite DNA that accounts for approx. 3% of the genome of the Bermuda land crab, Gecarcinus lateralis, was not amplified specifically. To establish optimal conditions for amplification of the segment of the G + C-rich satellite, we used two genetically engineered enzymes, AmpliTaq DNA polymerase and AmpliTaq DNA polymerase, Stoffel fragment (SF), and a number of denaturants or co-solvents. In the absence of denaturants or co-solvents, amplified products of both enzymes contained non-specific bands upon gel electrophoresis. Addition of certain denaturants or co-solvents to PCR mixtures resulted in the production of the single specific band of the expected size. Reagents that improved specificity of the amplified product were formamide, glycerol, DMSO, Tween-20 and NP-40; on the other hand, urea, ethanol and 1-methyl-2-pyrrolidone (NMP) inhibited amplification. Of the two enzymes, SF was more specific and efficient. The products of AmpliTaq DNA polymerase included one or more extra bands, even in the presence of denaturants or co-solvents, except for glycerol or DMSO.

Base Sequence↗

Survival strategies of plants associated with arbuscular mycorrhizal fungi on toxic mine tailings.

A field survey of metal concentrations and arbuscular mycorrhizal (AM) components of plants growing on five mining sites was conducted in Chenzhou City, Hunan Province, Southern China and a control site in Hong Kong. Significant differences were observed in the average concentrations of total heavy metals (Pb, Zn, Cu, Cd) and one metalloid (As) in contaminated soils compared with the control site. Gramineae and Compositae were the dominant plant families growing on mine tailings, with Chrysanthemum moritolium (common chrysanthemum), Cynodon dactylon (Bermuda grass), Miscanthus florodulus (Sword grass) and Pteris vittata (Ladder brake fern) commonly found at all sites. AM fungal colonization was detected in most of the plants. Comparing the four common plant species, three components of mycorrhizal colonization (arbuscules, vesicles and coiled hyphae) were found in the roots of C. dactylon and P. vittata growing at Do Shun Long (DSL) mine site. Concentrations of As in fronds were 24-fold higher than in roots of P. vittata with the highest mycorrhizal colonization rate (73%) among all sampling sites. Extensive mycorrhizal colonization (85%) was also recorded in the roots of C. dactylon with As accumulation 57 times higher than in shoots. The four common plants found in metal contaminated sites had developed different strategies for survival in the contaminated sites with the aid of indigenous AM fungi.

Arsenic↗