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Phormidium animalis (Cyanobacteria: Oscillatoriaceae) supports larval development of Anopheles albimanus.

The capability of Phormidium animalis, a cyanobacterium commonly found in larval habitats of Anopheles albimanus in southern Mexico, to support larval development of this mosquito was investigated. First-stage larvae were reared under insectary conditions with P. animalis ad libitum and their development was compared with larvae fed with wheat germ. The time of pupation and adult mosquito size, assessed by wing length, were similar in both groups, but fewer adult mosquitoes were obtained from larvae fed with the cyanobacteria. Nevertheless, these observations indicate that P. animalis is ingested and assimilated by larval An. albimanus, making this cyanobacterium a good candidate for genetic engineering for the introduction of mosquitocidal toxins for malaria control in the region.

Animals↗

Trimeric structure of human proliferating cell nuclear antigen. Implications for enzymatic function and autoantibody recognition.

The proliferating cell nuclear Ag (PCNA) is a DNA replication factor postulated to function as a sliding clamp around DNA. PCNA is also a target for autoimmunity in systemic lupus erythematosus. The autoantigenicity of PCNA is highly conformation-dependent, and reaction with most anti-PCNA sera requires a nearly full-length PCNA molecule. Here we describe the use of gel filtration and glycerol gradient sedimentation to analyze the native structure and size of PCNA. PCNA from three sources was studied (PCNA from HeLa cells, PCNA purified after its overexpression in bacteria, and PCNA produced in the wheat germ cell-free translation system) as well as mutant forms of PCNA translated in vitro. In each case, full-length PCNA behaved as a trimer. Analysis of mutant proteins revealed a correlation between the trimeric form and binding to the common type of human anti-PCNA autoantibody, suggesting that the Abs are specific for the active form of the protein. These findings are consistent with the idea that autoantibodies are generated as a response to native Ag and provide experimental support for the hypothesis that PCNA serves its processive function in DNA replication as a trimeric ring structure.

Autoantibodies↗

Functional and comparative bioinformatic analysis of expressed genes from wheat spikes infected with Fusarium graminearum.

Fusarium head blight, caused by the fungus Fusarium graminearum, is a major disease on wheat (Triticum aestivum L.). Expressed sequence tags (ESTs) were used to identify genes expressed during the wheat-F. graminearum interaction. We generated 4,838 ESTs from a cDNA library prepared from spikes of the partially resistant cultivar Sumai 3 infected with F. graminearum. These ESTs were composed of 2,831 singlet (single-copy transcripts) and 715 contigs (multiple-copy transcripts) for a total of 3,546 non-redundant sequences. Four sets of nonredundant sequences were identified. One set contains numerous, common biotic and abiotic stress-related genes. Many of these stress-related genes were represented by multiple ESTs, indicating that they are abundantly expressed. A second set comprised 16 nonredundant sequences from F. graminearum that may be required for pathogenicity. A subset of these fungal genes encodes proteins associated with plant cell wall degradation. A third set of 326 nonredundant sequences had no DNA or amino acid sequence similarity to almost 1 million plant and over 7 million animal sequences in dbEST (as of 22 June 2001). Thus, these 326 nonredundant sequences have only been found in our F. graminearum-infected 'Sumai 3' cDNA library. A fourth set of 29 nonredundant sequences was found in our F. graminearum-infected wheat and another plant-pathogen interaction cDNA library. Some of these sequences encode proteins that may act in establishing various plant-fungal interactions.

Cell Wall↗

High voltage electron microscopy of membrane interactions in wheat.

Structural interactions between intracellular membranes in root cells of Triticum sativum L. have been investigated with the 3 MeV electron microscope in Toulouse after selective labeling of the membranes with either zinc iodide-osmium tetroxide or phosphotungstate at low pH. Direct membrane continuities between endoplasmic reticulum, Golgi apparatus, and biogenetically derived compartments of the exoplasmic space are more extensive than commonly suggested from conventional electron microscopy. The results are discussed with reference to current concepts suggesting a membrane flow from the endoplasmic reticulum to the terminal components, plasmalemma, and vacuoles.

Endoplasmic Reticulum↗

Heterodera glycines: eggshell ultrastructure and histochemical localization of chitinous components.

The eggshell in most nematodes consists of an outer vitelline layer, a middle chitinous and an inner lipid layer. Earlier work with eggs of Heterodera glycines suggests the presence of two chitinous layers but the vitelline layer was not observed. From our observation the outer chitin layer described in past literature is actually a vitelline layer. Histochemical analysis has demonstrated that chitin is absent from the outer envelope. Electron microscope observations of the eggshell show a waxy appearance and osmium staining consistent with that of the proteinaceous vitelline layer found in other nematodes. Lectin localization also shows that the eggshell continues to develop past fertilization with the delivery and integration of eggshell precursors. Contrary to previous reports, we propose that the ultrastructure of the eggshell H. glycines follows the common three-layer structure observed in other nematodes.

Animals↗

Frequencies and sequence characteristics of di-, tri-, and tetra-nucleotide microsatellites in wheat.

Microsatellites have emerged as an important source of genetic markers for eukaryotic genomes. In this report, two wheat (Triticum aestivum L.) genomic libraries were screened for several di-, tri-, and tetranucleotide tandem repeats. Clones containing (AC)n, (AG)n, (TCT)n, and (TTG)n repeats were isolated and sequenced. On average, there was one (AC)n microsatellite every 292 kbp and one (AG)n microsatellite every 212 kbp. The trinucleotide tandem repeats (TCT)n and (TTG)n were about 10 times less common than the two dinucleotide tandem repeats tested and tetranucleotide tandem repeats were rare. Many of the microsatellites had more than 10 repeats. The maximum repeat number found for (AC)n was 36 and for (TCT)n was more than 50. The prevailing category of (AG)n microsatellites from (AG)n isolates was perfect repeats. About half of the (AC)n microsatellites were compound repeats, while most of the (TCT)n microsatellites were imperfect repeats. In a small sample, (TTG)n microsatellites consisted mainly of compound repeats. The most frequently associated repeats were (AC)n with (AG)n, (TCT)n with (TCC)n, and (TTG)n with (TGG)n. Among 32 pairs of microsatellite primers surveyed, seven produced polymorphic products in the expected size range and these loci were mapped using a hexaploid wheat mapping population or aneuploid stocks.

Base Sequence↗

Immunochemical studies of Aspergillus fumigatus mycelial antigens by polyacrylamide gel electrophoresis and western blotting techniques.

Differences were detectable among strains of the opportunist fungal pathogen Aspergillus fumigatus when water-soluble (WS) preparations were analysed by combined SDS-PAGE and Western blotting procedures. A wide range of molecules of apparent molecular masses from approximately 20 to greater than 100 kDa showed specific binding to antibodies raised in rabbits to A. fumigatus wall and cytoplasmic components. The ability to bind antibody was markedly reduced by treatment of these antigens with sodium periodate or with specific proteases or glucanases. Pretreatment of blotted antigens with either concanavalin A (ConA) or wheat germ agglutinin (WGA) did not, however, inhibit subsequent antibody binding. The antigens of subfractions prepared from a single strain of A. fumigatus WS material were also susceptible to periodate oxidation and enzymic hydrolysis. Slight cross-reactivity was apparent when crude preparations of cellular or culture filtrate antigens, used in this laboratory to detect antibodies to Candida albicans, Coccidioides immitis and Cryptococcus neoformans, were probed with hyperimmune rabbit antisera to A. fumigatus. Efforts were made to characterize the WS preparations of A. fumigatus, used as diagnostic antigens in many laboratories. The electrophoretically separated antigenic moieties were shown to be predominantly glycoproteins. Binding of cytoplasmic antigens to antibodies raised to wall material showed the presence of many common components in both wall and cytosol. Antiserum to wall components revealed most differentiation among A. fumigatus strains.

Antigens, Fungal↗

Fungal flora of poultry feedstuff ingredients.

One hundred and ten samples representing five types of poultry feed ingredients were mycologically examined. These samples included soybean meal, ground maize, cotton-seed cake, wheat bran and fish meal (22 samples each). Among the 73 mesophilic fungal species, Aspergillus flavus, A. niger and A. fumigatus were the most dominant. A. terreus, A. flavipes, Mucor circinelloides, Scopulariopsis brevicaulis, Penicillium chrysogenum, Fusarium moniliforme and Rhizopus stolonifer were found to be common on a particular ingredient and less common on the remainders. Of the twelve thermophilic and thermotolerant species, A. fumigatus, Thermomyces lanuginosus, Rhizomucor pusillus and Thermoascus thermophilus prevailed on one or more type of the different ingredients. Marked variations were observed in the rancid fatty compounds of the different samples and the values ranged between 0.663 and 3.900 mg malonaldehyde per kg sample.

Animal Feed↗

Contribution to the fungal flora of cereal grains in Egypt.

30 genera and 77 species, in addition to 4 varieties were isolated from 25 samples of each of barley, wheat, maize and sorghum grains collected from different places in Egypt. The broadest spectrum of genera and species was recorded in wheat (25 genera and 59 species + 4 varieties) followed by barley (21 genera and 52 species + 2 varieties), sorghum (14 genera and 33 species + 2 varieties) and maize grains (11 genera and 29 species + 2 varieties). Aspergillus, Fusarium, Penicillium, and Rhizopus were the most common genera in the four grains, except maize where Penicillin emerged in low frequency. Aspergillus was the main component of the fungal flora of the four grains and contributed 79-94.4% of the gross total count of fungi. From the preceding genera A. niger, A. flavus, A. fumigatus, F. oxysporum, P. chyrsogenum, P. corylophilum, P. notatum and R. stolonifer were the most frequent.

Aspergillus↗

Cytology, synaptology and immunocytochemistry of commissural neurons and their putative axonal terminals in the dorsal cochlear nucleus of the rat.

The first binaural integration within the auditory system responsible for sound localization depends upon commissural neurons that connect the two symmetrical cochlear nuclei. These cells in the deep polymorphic layer of the rat dorsal cochlear nucleus were identified with the electron microscope after injection of the retrograde tracer, Wheat Germ Agglutinin conjugated to Horseradish Peroxydase, into the contralateral cochlear nucleus. Commissural neurons are multipolar or bipolar with an oval to fusiform shape. Few commissural neurons, most inhibitory but also excitatory, connect most of the divisions of the rat cochlear nuclei. The most common type is a glycinergic, sometimes GABAergic, moderately large cell. Its ergastoplasm is organized into peripheral stacks of cisternae, and few axo-somatic synaptic boutons are present. Another type of commissural neuron is a medium-sized, spindle-shaped cell, glycine and GABA-negative, with sparse ergastoplasm and synaptic coverage. A giant, rare type of commissural neuron is glycine-positive and GABA-negative, with short peripheral stacks of ergastoplasmic cisternae. It is covered with synaptic boutons, many of which contain round synaptic vesicles. Another rare type of commissural neuron is a moderately large cell, oval to fusiform in shape, immunonegative for both glycine and GABA, and contacted by many axo-somatic boutons. It contains large dense mitochondria and numerous dense core vesicles of peptidergic type. Some labelled boutons, mostly inhibitory and probably derived from commissural neurons, contact pyramidal, cartwheel, giant and tuberculo-ventral neurons. The prevalent inhibition of electrical activity in a cochlear nucleus observed after stimulation of the contralateral cochlear nucleus may be due to commissural inhibitory terminals which contact excitatory neurons such as pyramidal and giant cells. Other inhibitory commissural terminals which contact inhibitory neurons such as cartwheel and tuberculo-ventral neurons, may explain the stimulation of electrical activity in the DCN after contralateral stimulation.

Animals↗

Life- cycle assessment in pesticide product development: methods and case study on two plant-growth regulators from different product generations.

Environmental assessments in pesticide product development are generally restricted to plant uptake and emissions of active ingredients. Life-cycle assessment (LCA) enables a more comprehensive evaluation by additionally assessing the impacts of pesticide production and application (e.g. tractor operations). The use of LCA in the product development of pesticides, in addition to the methods commonly applied, is therefore advisable. In this paper a procedure for conducting LCA in early phases of product development is proposed. In a case study, two plant-growth regulators from different product generations were compared regarding their application in intensive production of winter wheat. The results showed thatthe reduced emissions from active ingredients of the newer pesticide were compensated by higher impacts from the production process. The authors draw the conclusion that it is important to consider environmental objectives in the procurement of precursors, in addition to the classical goals of increasing the efficacy and reducing the nontarget effects of pesticides. Moreover, the case study showed that decisions based on uncertain results in early stages of product development may need to be revised in later stages, e.g. based on investigations of pesticides' effects on crop yield.

Chlormequat↗

Ingredient and labeling issues associated with allergenic foods.

Foods contain a wide range of food ingredients that serve numerous technical functions. Per capita consumer exposure to most of these food ingredients is rather low with a few notable exceptions such as sugar and starch. Some food ingredients including edible oils, hydrolyzed proteins, lecithin, starch, lactose, flavors and gelatin may, at least in some products, be derived from sources commonly involved in IgE-mediated food allergies. These ingredients should be avoided by consumers with allergies to the source material if the ingredient contains detectable protein residues. Other food ingredients, including starch, malt, alcohol and vinegar, may be derived in some cases from wheat, rye or barley, the grains that are implicated in the causation of celiac disease. If these ingredients contain gluten residues, then they should be avoided by celiac sufferers. A few food ingredients are capable of eliciting allergic sensitization, although these ingredients would be classified as rarely allergenic. These ingredients include carmine, cochineal extract, annatto, tragacanth gum and papain. Food manufacturers should declare the presence of allergenic food ingredients in the ingredient listings on product labels so that allergic consumers can know to avoid these potentially hazardous products.

Allergens↗

A large product of cell-free translation of messenger RNA coding for corticotropin.

Polyadenylate-containing RNA prepared from the membrane fraction of bovine anterior pituitary gland was shown to direct the synthesis of a large translation product related to corticotropin (adrenocorticotropic hormone) in a cell-free system derived from wheat germ. This product was identified by immunoprecipitation with specific antibody against corticotropin, followed by electrophoretic analysis of the dissociated immunoprecipitate on sodium dodecyl sulfate-polyacrylamide gel. Further evidence for the identity of the translation product was provided by the presence of a common peptide in the chymotryptic digest of [35S]methionine-labeled cell-free product and in that of authentic corticotropin. The molecular weight of the translation product was estimated to be approximately 35,000, based on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. These results indicate that corticotropin messenger RNA directs the cell-free synthesis of a product that contains the amino acid sequence of corticotropin but is much larger than this hormone.

Adrenocorticotropic Hormone↗

Morphogenesis in germinating Fusarium graminearum macroconidia.

Fusarium graminearum (teleomorph Gibberella zeae) is a significant pathogen of wheat and corn. F. graminearum forms multicellular macroconidia that play an important role in dissemination of the disease. The spatial pattern of morphogenesis in germinating macroconidia is described. Germ tubes preferentially emerge from the apical cells in a bipolar pattern that appears to be common to filamentous fungi. Chitin deposition occurs at two locations: the spore apices and cortical regions of macroconidial cells that subsequently produce a germ tube. The spatial pattern of morphogenesis requires the presence of functional microtubules, which may be responsible for the transport of key polarity factors to specific sites. These observations suggest that F. graminearum possesses a regulatory system that marks germ tube emergence sites. Perturbation of this system may represent an effective approach for inhibiting colonization of host plant surfaces.

Cell Polarity↗

An epidemiological study of precursor lesions of esophageal cancer among young persons in a high-risk population in Huixian, China.

The occurrence of chronic esophagitis, considered a precursor condition for esophageal cancer, among persons 15 to 26 yr of age and risk factors for the disease were investigated in Huixian, Henan Province, a high-risk area for esophageal cancer in the People's Republic of China. The 538 study subjects underwent an esophagoscopy with guided biopsies and cytology, a physical examination, an interview with a questionnaire including known and suspected risk factors for esophageal lesions, and collection of a 10-ml blood sample and overnight urine. One-third of the subjects was selected from households with a case of esophageal cancer in the past 6 yr and two-thirds came from control households. Histologically confirmed very mild, mild, and moderate esophagitis was observed in 31.6%, 10.7%, and 1.1% of 354 male and 30.4%, 4.3%, and 1.1% of 184 female subjects, respectively. In the multivariate case-control analysis of mile and moderate esophagitis compared with very mild esophagitis and normal subjects, the prevalence of mild and moderate disease was found to be positively associated with the consumption of burning hot beverages [odds ratio (OR) = 4.7], the prevalence of esophagitis among siblings (OR = 4.4), and family history of esophageal cancer (OR = 1.8) and negatively associated with the frequent consumption of fresh fruits (OR = 0.3) and wheat flour products (OR = 0.4). Weaker associations were seen for cigarette smoking and the use of cottonseed oil as the main cooking oil. Univariate associations seen with a clinical diagnosis of oral leukoplakia (OR = 2.7) and seborrheic dermatitis (OR = 3.7) are probably due to common risk factors such as smoking and nutritional deficiency. The present findings suggest that exposures early in life to environmental risk factors and nutritional deficiency may be responsible for inflammation and a weakened esophageal epithelium, resulting in a condition possibly more favorable for the development of esophageal cancer.

Adolescent↗

Diagnostic value of alkaline phosphatase isoenzyme separation by affinity electrophoresis in the dog.

Affinity electrophoresis, using wheat germ lectin, was used to separate the alkaline phosphatase isoenzymes in the sera of 150 dogs with alkaline phosphatase values greater than or equal to 150 IU/L. The method provided clearer separation of the liver, bone and steroid-induced alkaline phosphatase isoenzymes commonly observed in canine serum, compared to conventional cellulose acetate electrophoresis. The dogs were divided into four patient groups determined by previous corticosteroid treatment, evidence of elevated endogenous corticosteroid levels, age and alanine aminotransferase values. The isoenzyme pattern of each patient was qualitatively assessed. The isoenzyme pattern most frequently observed was greater than 50% steroid induced alkaline phosphatase, which was present in 76 of 150 dogs. This pattern was observed in 18 of 22 dogs receiving corticosteroid therapy, two of three dogs with hyperadrenocorticism, and in dogs with a variety of other diagnoses. The majority of immature dogs (12 of 20) had an isoenzyme pattern consisting of greater than 50% bone. The majority of dogs with active hepatocellular injury (16 of 27) had greater than 50% liver isoenzyme. The isoenzyme pattern was not specific for certain diseases, therefore the diagnostic usefulness is limited. However the isoenzyme result is useful in some cases to determine which further diagnostic tests are indicated, and to determine the source of alkaline phosphatase elevation.

Adrenal Cortex Hormones↗

Surgical case of aortic root and thoracic aortic aneurysm after the Wheat procedure.

A 52 year-old man underwent aortic valve replacement and ascending aortic replacement (Wheat procedure) for acute dissection (Stanford type A) and aortic regurgitation (grade 3/4). At that time, the aortic root was slightly dilated at about 45 mm and the descending aorta was within a normal range at about 35 mm. Forty months after the initial operation, a follow-up chest enhanced computed tomography showed an aortic root aneurysm about 60 mm in diameter, a thoracic aortic aneurysm about 70 mm in diameter and chronic aortic dissection. First we performed the Bentall procedure, innominate artery and left common carotid artery replacement by 12 mm, and 10 mm Hemashield grafts during selective cerebral perfusion. After 10 weeks, we carried out aortic arch, descending aorta and left subclavian artery replacement. The postoperative course was uneventful and postoperative examination demonstrated a good surgical result. Histological findings of the aortic aneurysm wall showed cystic medial necrosis, but Marfan's syndrome was excluded clinically. We could diagnose aortic root aneurysm by regular follow-up chest enhanced computed tomography (CT) and echocardiography. Therefore, cases with slight dilation of the aortic root in the Wheat procedure should undergo regular follow-up evaluation by chest enhanced CT and echocardiography.

Aortic Dissection↗

Urinary bladder innervation in male rat: termination of primary afferents in the spinal cord as determined by transganglionic transport of WGA-HRP.

The distribution of afferents innervating the urinary bladder in the spinal cord of male rats has been studied with the axonal tracer horseradish peroxidase conjugated to wheat germ agglutinin (WGA-HRP) injected into various portions of one side of the urinary bladder (dome, body, base, or neck) and other pelvic organs (prostate and rectum). Labeled neurons were found in dorsal root ganglia of the lumbosacral cord (L1-S3, peak in S1-S2). The strongest and most extensive transganglionic labeling of primary afferents resulted after injections in the body of the bladder. Primary afferents were observed bilaterally in Lissauer's tract and laminae I-II at the apex of the dorsal horn, from L6 to S3. The projection extended laterally up to the sacral parasympathetic nucleus and medially up to the gray matter dorsal to the central canal, where they formed a plexus of fibers and terminals. Deposits in the dome and base of the bladder labeled more heavily the medial projection, while the least intense projection was seen after injections in the bladder neck. Our results indicate a common pattern of termination of primary afferents from the bladder, although some topographical differences exist.

Afferent Pathways↗