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[Tolerance and digestibility of Peruvian "common foods" in malnourished infants].

Six diets were prepared based on commonly used Peruvian foods, mainly of vegetable origin, which were offered to eight infants (mean age: 12.8 +/- 8.2 months) recovering from malnutrition. The purpose of this study was to evaluate the acceptability, tolerance and digestibility of the diets in question. Five were prepared with a potato and wheat base (noodle) and the sixth with a quinua-oats base. The acceptability and tolerance was satisfacotry, allowing maintenance of an adequate calorie and protein intake in all patients except one. On the quinua-oats based diet, the mean apparent absorption of nitrogen and fat was significantly lower (P less than 0.05) than in the case of the other diets. The increase in height coefficient (height age/chronological age X 100) and weight/age proved to be adequate during the study, except in the three youngest patients. The authors consider that this type of diets (potato-wheat based) can be recommended for infant feeding after the first year of life, and that the quinua-oats based diet still needs a more thorough evaluation prior to recommending its use.

Body Weight↗

Lectins for the identification of ocular bacterial pathogens.

In a preliminary in vitro investigation, fluorescein-conjugated lectins were used in the identification of bacteria commonly involved in ocular infections. Clinical isolates of Staphylococcus aureus, Staph. epidermidis, Streptococcus pneumoniae, Strep. pyogenes, Pseudomonas aeruginosa, Hemophilus influenzae, and Proteus mirabilis were incubated with each of eleven lectins using a slide technique. Bacterial fluorescence was readily observed with a fluorescence microscope. All clinical isolates bound wheat germ agglutinin. The Gram-positive isolates bound Concanavalin A, while the Gram-negative isolates did not, with rare exceptions. Streptococcal species isolates reacted with Dolichos biflorus agglutinin, while staphylococcal species isolates did not. Lectins may be useful in furthering the initial identification of causative organisms in bacterial ocular infections.

Bacterial Infections↗

Development and organization of ocular dominance bands in primary visual cortex of the sable ferret.

Thalamocortical afferents in the visual cortex of the adult sable ferret are segregated into eye-specific ocular dominance bands. The development of ocular dominance bands was studied by transneuronal labeling of the visual cortices of ferret kits between the ages of postnatal day 28 (P28) and P81 after intravitreous injections of either tritiated proline or wheat germ agglutinin-horseradish peroxidase. Laminar specificity was evident in the youngest animals studied and was similar to that in the adult by P50. In P28 and P30 ferret kits, no modulation reminiscent of ocular dominance bands was detectable in the pattern of labeling along layer IV. By P37 a slight fluctuation in the density of labeling in layer IV was evident in serial reconstructions. By P50, the amplitude of modulation had increased considerably but the pattern of ocular dominance bands did not yet appear mature. The pattern and degree of modulation of the ocular dominance bands resembled that in adult animals by P63. Flat mounts of cortex and serial reconstructions of layer IV revealed an unusual arrangement of inputs serving the two eyes in the region rostral to the periodic ocular dominance bands. In this region, inputs serving the contralateral eye were commonly fused along a mediolateral axis, rostral to which were large and sometimes fused patches of ipsilateral input.

Aging↗

Topographic organization of rat locus coeruleus and dorsal raphe nuclei: distribution of cells projecting to visual system structures.

Previous reports from this laboratory and elsewhere have provided evidence that the locus coeruleus (LC) and dorsal raphe (DR) nuclei are topographically organized with respect to their efferent targets. Whereas most of these previous studies have focused on relationships between these monoamine-containing brainstem nuclei and cerebral cortex, basal ganglia, and limbic structures, they have not systematically examined the distribution of LC and DR cells that project to multiple structures with common sensory or motor functions. The goal of the present study was to characterize and compare the distributions of LC and DR cells which project to different visual areas of the rat central nervous system. Long-Evans hooded rats received unilateral pressure injections of the retrograde tracer wheat germ agglutinin-horseradish peroxidase in either the dorsal lateral geniculate, ventral lateral geniculate, or lateral posterior nucleus of thalamus; superior colliculus, cortical area 17, cortical area 18a/b; cerebellar vermis (lobules VI and VII); or paraflocculus. Transverse sections through the midbrain and pons were examined by light microscopy after performing routine tetramethyl benzidine histochemical procedures. For all cases studied, retrogradely labeled cells were observed throughout the rostrocaudal extent of the LC and DR; however, labeling patterns which were distinctive for different injection sites were noted in each of these brainstem nuclei. The major conclusion drawn from this work is that subsets of LC and DR cells which project to different target structures within the rat visual system are found in overlapping but not necessarily coextensive zones within these nuclei. These studies provide further evidence of a rough topographic ordering within both the LC and DR nuclei, as well as support a new hypothesis that the outputs from each of these nuclei are organized with respect to the sensory related functions of their efferent targets.

Animals↗

A form of muco-cutaneous leishmaniasis in the Old World.

Four Ethiopian patients with leshmaniasis affecting the nose or the mouth are described. Leishmaniasis affecting these sites is relatively common in Ethiopia; this resembles South American leishmaniasis but probably results from direct extension from skin lesions rather than from metastatic spread of organisms.

Adolescent↗

Fast temperature programming in routine analysis of multiple pesticide residues in food matrices.

Flash gas chromatographic (GC) analysis of 15 organophosphorus pesticides commonly occurring in food crops was performed using the Thermedics Detection EZ Flash upgrade kit installed in the oven of a HP 5890 Series II Plus gas chromatograph. The temperature program and splitless time period were the main parameters to be optimized. In the first set of experiments wheat matrix-matched standards were analyzed both by: (i) the flash GC technique (resistive heating of a 5 m capillary column), and (ii) the conventional GC technique (moderate oven temperature programming of a 30 m capillary column). Using the flash GC technique, the analysis time was reduced by a factor of more than 10 compared to the conventional GC technique. Dramatically improved detectability of analytes was achieved due to much narrower peak widths. The flash GC technique was compared with another approach to faster GC analysis employing a 5 m column and fast temperature programming with a conventional GC oven. In comparison with this alternative, in the case of flash GC significantly better retention time repeatability was observed. The other superiority of resistive heating is very rapid cooling down (i.e., equilibration to the initial conditions) which contributes to the increased sample throughput.

Chromatography, Gas↗

Clinical and histopathologic features of corneal dystrophies in Japan.

PURPOSE: To examine retrospectively the frequency of various corneal dystrophies among Japanese patients who underwent keratoplasty or keratectomy at the authors' institution over a 34-year period, and to compare the histopathologic features of these disorders in the Japanese population with those reported in the Western literature. METHODS: Corneal specimens obtained during keratectomy or keratoplasty (lamellar and penetrating) performed at the authors' institution from 1959 through 1992 were reviewed. Immunohistochemical studies were performed using monoclonal antibodies to keratan sulfate and gelsolin, as well as two lectins (concanavalin A and wheat germ agglutinin). RESULTS: Of 1259 corneal specimens, 159 (12.6%) specimens from a total of 80 patients showed corneal dystrophy. Virtually all were non-Fuchs dystrophies; only one case of primary Fuchs dystrophy was identified histologically. Granular dystrophy and gelatinous drop-like dystrophy were the most common dystrophies identified in the specimens, largely because of multiple specimens from individual patients with recurrent disease. These two disorders accounted for 86 of the 159 specimens. In terms of numbers of patients, lattice dystrophy was the most common (26 patients, 32.5%), followed by macular dystrophy (16 patients, 20%), gelatinous drop-like dystrophy (15 patients, 18.8%), granular dystrophy (14 patients, 17.5%), and Avellino dystrophy (3 patients, 3.75%). Dystrophies represented by only one or two patients included congenital hereditary endothelial dystrophy, primary spheroidal keratopathy, posterior polymorphous dystrophy, Schnyder crystalline dystrophy, and Fuchs dystrophy. CONCLUSIONS: This histopathologic study showed a very low incidence of Fuchs dystrophy in the authors' Japanese patient population, compared with the incidences seen in studies of populations in Western countries. Of the non-Fuchs dystrophies, lattice dystrophy was the most common among the patients, although there were large numbers of specimens with granular dystrophy and gelatinous drop-like dystrophy due to their recurrent character. The causes of clinical and histopathologic differences and similarities among the Japanese patients and the patients described in the Western literature are likely related to genetic factors, but a complete understanding of their specific mechanisms awaits future molecular biologic and genetic elucidation.

Antibodies, Monoclonal↗

Genomic changes in synthetic Arabidopsis polyploids.

Polyploids are common and arise frequently by genome duplication (autopolyploids) or interspecific hybridization (allopolyploids). Neoallopolyploids display sterility, lethality, phenotypic instability, gene silencing and epigenetic changes. Little is known about the molecular basis of these phenomena, and how much genomic remodeling happens upon allopolyploidization. Extensive genomic remodeling has been documented in wheat, but little remodeling occurs in cotton. Newly synthesized Arabidopsis allopolyploids, which display phenotypic instability and low fertility, displayed several, possibly related mechanisms that can remodel genomes. We detected transcriptional activity of several transposons although their transposition was limited. One represents a new family of conditionally active En-Spm-like transposons of Arabidopsis thaliana, which underwent remodeling of CG methylation upon allopolyploidization. A random amplified fragment length polymorphism survey suggested remodeling at few, specific loci. Meiotic analyses revealed the appearance of chromosomal fragments in a substantial fraction of anther meiocytes. In several individuals produced by hybrids between the synthetic and a natural allopolyploid pollen viability inversely correlated with meiotic instability. Activity of selected DNA transposons and the possibly related chromosomal breaks could cause changes by inducing translocations and rearrangements.

Arabidopsis↗

A unique fluorescent phenylalkylamine probe for L-type Ca2+ channels. Coupling of phenylalkylamine receptors to Ca2+ and dihydropyridine binding sites.

The first fluorescently labeled phenylalkylamine, DMBODIPY-PAA (5-(3-[3-(4,4-difluoro-5,7-dimethyl-3a, 4a-diaza-4-bora-indacen-3-yl)propionamido] phenethyl-N-methylamino)-2-isopropyl-2-(3,4,5-trimethoxyphenyl)-valer onitrile) has been introduced for L-type Ca2+ channel research. DMBODIPY-PAA binds reversibly to L-type Ca2+ channels purified from rabbit skeletal muscle microsomes by wheat germ agglutinin-Sepharose chromatography. In this preparation DMBODIPY-PAA labels 412 pmol of phenylalkylamine receptors/mg of protein with a Kd of 6.82 nM and a favorable signal-to-noise ratio. Therefore DMBODIPY-PAA has a higher affinity for purified Ca2+ channels than the commonly employed radioligands and consequently has assisted in channel purification after prelabeling by simply monitoring receptor-bound fluorescence. (+)-PN200-110 (which is stimulatory for (-)-[3H]desmethoxyverapamil binding to purified Ca2+ channels) inhibits DMBODIPY-PAA labeling. Since these drug interactions are reciprocal, the phenylalkylamine and dihydropyridine binding sites of the alpha 1-subunit are tightly coupled. Kinetic and equilibrium binding studies with (-)-[3H]desmethoxyverapamil and DMBODIPY-PAA show that phenylalkylamine binding to L-type Ca2+ channels is dependent on Ca2+. Chelation of divalent metal ions converts phenylalkylamine receptors into a very low affinity state. This conversion is temperature- and time-dependent and completely reversible (K0.5 for free Ca2+ = 58 nM). This study demonstrates the utility of fluorescent ligands for binding studies with L-type Ca2+ channels and provides evidence for coupling between Ca2+ binding sites and phenylalkylamine receptors.

Animals↗

The origin of corticospinal projections from the premotor areas in the frontal lobe.

We determined the origin of corticospinal neurons in the frontal lobe. These neurons were labeled by retrograde transport of tracers after injections into either the dorsolateral funiculus at the second cervical segment or the gray matter of the spinal cord throughout the cervical enlargement. Using retrograde transport of tracer from the arm area of the primary motor cortex, we defined the arm representation in each premotor area in another set of animals. We found that corticospinal projections to cervical segments of the spinal cord originate from the primary motor cortex and from the 6 premotor areas in the frontal lobe. These are the same premotor areas that project directly to the arm area of the primary motor cortex. The premotor areas are located in parts of cytoarchitectonic area 6 on the lateral surface and medial wall of the hemisphere, as well as in subfields of areas 23 and 24 in the cingulate sulcus. The total number of corticospinal neurons in the arm representations of the premotor areas equals or exceeds the total number in the arm representation of the primary motor cortex. The premotor areas collectively comprise more than 60% of the cortical area in the frontal lobe that projects to the spinal cord. Like the primary motor cortex, each of the premotor areas contains local regions that have a high density of corticospinal neurons. These observations indicate that a substantial component of the corticospinal system originates from the premotor areas in the frontal lobe. Each of the premotor areas has direct access to the spinal cord, and as a consequence, each has the potential to influence the generation and control of movement independently of the primary motor cortex. These findings raise serious questions about the utility of viewing the primary motor cortex as the "upper motoneuron" or "final common pathway" for the central control of movement.

Afferent Pathways↗

Identification of a cell surface glycoprotein family of olfactory receptor neurons with a monoclonal antibody.

A monoclonal antibody (Mab) has been developed which recognizes a family of cell surface glycoproteins found in high levels of rat olfactory receptor neurons. This Mab, designated 2B8, was produced by the fusion of X63-Ag8.653 myeloma cells and spleen cells of a mouse immunized with PC12 rat pheochromocytoma cells. Immunofluorescence analyses of cryostat sections of neonatal olfactory epithelium show prominent 2B8 binding to receptor neurons. Within the olfactory bulb only the glomerular and olfactory nerve layers show 2B8 binding. All other neural structures in the main olfactory bulb have background levels of reactivity. Analyses of 2B8 binding to particulate protein preparations from several central and peripheral nervous system components demonstrated highest 2B8 antigen specific activity in olfactory bulb and epithelium and detectable levels in dorsal root ganglia (DRG), whole cerebrum, cerebellum, and brainstem. However, 2B8 antigen could not be detected in non-olfactory structures by immunofluorescence. Some non-neural tissues also had the ability to bind 2B8 Mab in the particulate protein radioimmunoassay. In order to compare the 2B8-reactive molecules found in each tissue, Mab was applied to polyacrylamide gels of unlabeled membrane proteins. A family of molecules with diverse molecular weights was found. Some were unique to individual tissues whereas others were shared among tissues. Olfactory bulb and epithelium had a unique band with Mr = 215,000 and another band with Mr = 142,000. The 142,000-dalton band was also found with PC12 cells. PC12 cells also had several bands of lesser molecular weight, including 51,000 and 43,000. Testes membranes had immunoreactive bands only at Mr = 46,000 and 43,000. Bone marrow, perinatal liver, and DRG each expressed a single 2B8-reactive band with Mr = approximately 114,000. Salivary gland had four reactive bands, two common to it and only PC12 cells, the 114,000-dalton band which is similar to that found in adult rat bone marrow and DRG, and a unique band at Mr = 152,000. 2B8 immunoprecipitates of olfactory bulb and epithelium were analyzed for glycosyl groups by lectin reactivity. Wheat germ agglutinin and Ricinus communus agglutinin I bound the 2B8 antigens using two distinct assay methods. This suggests that the 2B8 antigens recognized in the olfactory system are glycoproteins having sialic acid and D-galactosyl components.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Inhalant allergens: as a cause of respiratory allergy in east Mediterranean area, Turkey.

Inhalant allergens were identified by analyzing the positive skin tests of 614 respiratory allergic patients. The important indoor allergens causing skin test positivity were house dust, Dermatophogoides pteronyssinus, cockroach, mosquito and Dermatophogoides farinea. Johnson grass, cultivated wheat, rye, orchard, and cultivated corn were among important grass pollens. Animal allergens like cow hair, cat and dog dander, bird and chicken feathers and horse hair caused skin test positivity in our allergic patients. Weeds and garden plants were also among important inhalant allergens in this region. Common cocklebur, narcissus, rose, mugworth, and daisy caused skin test positivity in decreasing frequencies. Carob, willow, poplar, olive and cypress pollens were among important inhalant allergens causing skin test positivity that may be important cause of respiratory allergy in this region.

Adolescent↗

Study of IgE antigenic relationships in hypersensitivity to hydrolyzed wheat proteins and wheat-dependent exercise-induced anaphylaxis.

BACKGROUND: Wheat is involved in different forms of respiratory, food and contact allergy. The IgE of patients generally reacts with various flour proteins. It is not known if antigenic relationships could explain some of these reactions and if proteins could be involved in different pathologies. METHODS: Two sera were selected as representative of patients with either wheat-dependent exercise-induced anaphylaxis (WDEIA) or hypersensitivity to hydrolyzed wheat proteins (HHWP). Their IgE specificity was studied with wheat, barley and rye proteins, using immunoblot, and immunoblot inhibition with recombinant gamma-3 hordein. This protein was chosen for its cross-reactivity with omega-5 gliadin, a major allergen in WDEIA. RESULTS: The IgE from both sera strongly reacted with natural and recombinant gamma-3 hordein but displayed different patterns of reactivity with wheat, barley and rye proteins. Those from the WDEIA patient showed expected reactions with omega-5 gliadin, gamma-35 and gamma-75 secalins, but also with wheat low-molecular-weight glutenin subunits (LMW-GS), and not with C hordeins. On the contrary, IgE from a HHWP patient reacted with C hordeins, various omega gliadins, and gamma-75 secalin, but very weakly with gamma-35 secalin and LMW-GS. Recombinant gamma-3 hordein inhibited strongly but not totally the WDEIA patient's IgE binding to prolamins. No such inhibition could be observed for the HHWP patient's IgE. CONCLUSIONS: At least part of the reactions of prolamins with the IgE from the WDEIA patient was due to antigenic homologies. The occurrence of cross-reacting carbohydrates was unlikely. These common IgE epitopes were not involved in the pathology of the HHWP patient.

Adult↗

Ipsilateral cortical connections of granular frontal cortex in the strepsirhine primate Galago, with comparative comments on anthropoid primates.

Modern studies of granular frontal cortex (GFC) in large-brained, anthropoid primates, such as Macaca, indicate that this region is comprised of many areal subdivisions. These areas vary in their architectonic appearance and each has a distinctive, diverse set of corticocortical connections. The great extent of the GFC region in anthropoids, and its high degree of areal parcellation, suggest that some GFC areas may be specializations of anthropoids, not found in other mammals. To investigate this possibility, we studied the corticocortical connections of GFC in the relatively small-brained, strepsirhine primate Galago, with a series of eight tracer injections in the frontal cortex, and an additional eight injections of parietal and temporal cortex. Tracers used were wheat-germ agglutinin conjugated to horseradish peroxidase and tritiated amino acids. Our results indicate that Galago GFC has strong, reciprocal connections with the parietal area-7 complex and with higher-order temporal areas; there are additional connections with extrastriate visual cortex, parahippocampal, and cingulate areas, and frontal cortex. Thus GFC has an extremely diverse array of cortical connections in Galago, as in Macaca. However, we also found that the pattern of parietofrontal connections is simpler in Galago than in Macaca. Specifically, parietal areas project to fewer discrete zones within the GFC of Galago, consistent with the view that these animals have fewer GFC areas than Macaca. In addition, Galago GFC possesses connections that specifically resemble those of Macaca arcuate cortex, but lacks connectional patterns that are characteristic of principalis cortex. These results are in accord with our previous architectonic studies, which indicated that Galago does not possess homologues of principalis areas. We conclude that the arcuate areas are common elements of primate GFC organization, while the areas located within and adjacent to the principal sulcus are anthropoid specializations.

Animals↗

Hamster hepatic cytochrome b5: purifications, immunochemical properties, and in vitro synthesis.

Cytochrome b5 has been purified from hamster liver microsomes. Both Ouchterlony double-diffusion and rocket immunoelectrophoresis experiments indicate that no immuno-cross-reactivity exists between guinea-pig anti-rabbit cytochrome b5 antibody and hamster cytochrome b5. However, anti-rabbit b5 IgG inhibited both hamster microsomal NADH-cytochrome c reductase and NADPH-dependent 7-ethoxycoumarin-O-deethylase activities. Hamster cytochrome b5 stimulated several reconstituted hamster cytochrome P-450-dependent monooxygenase activities and this stimulatory effect could be inhibited by antibody against rabbit cytochrome b5. Two-dimensional iodinated tryptic peptide mapping experiments provided evidence that the polypeptide fingerprint of hamster cytochrome b5 is substantially different from the fingerprints of cytochrome b5 isolated from rabbit, rat and bovine. We also studied the in vitro synthesis of hamster cytochrome b5 from liver mRNA using a wheat germ lysate system. A 16 kDa polypeptide, which is the same size as hamster cytochrome b5, was immunoprecipitated by antibody against rabbit b5. This experiment suggested that in vitro synthesized hamster cytochrome b5 is recognized by a heterologous antibody. Thus, hamster and rabbit cytochrome b5 do share some common immuno-determinants which may be located close to the heme-binding active site.

Animals↗

Biogenesis of the platelet receptor for fibrinogen: evidence for separate precursors for glycoproteins IIb and IIIa.

Congenital absence of platelet glycoproteins IIb and IIIa (GPIIb and GPIIIa) results in a severe bleeding disorder characterized by defective platelet aggregation and failure of fibrinogen to bind to platelets. GPIIb is a two-chain protein containing disulfide-linked alpha and beta subunits. GPIIb and GPIIIa are present as a heterodimeric, noncovalent complex in the platelet plasma membrane and function as the fibrinogen receptor. To characterize synthesis of these two proteins, RNA isolated from a human leukemia cell line that contains GPIIb and GPIIIa was translated in a wheat germ cell-free system. Polyclonal antibodies specific for each protein immunoprecipitated distinct [35S]methionine-labeled precursors, indicating that GPIIb and GPIIIa are translated from separate mRNAs. Moreover, using specific antibodies against either intact unreduced GPIIb or the beta subunit, we obtained evidence for synthesis of a common polypeptide precursor for GPIIb alpha and GPIIb beta. Based on experiments using microsomal membranes, it appears that GPIIb is integrated into the platelet membrane with little or no cytoplasmic component. These results suggest that precursors of GPIIb and GPIIIa may be encoded by separate genes and that each precursor is processed before delivery to the plasma membrane.

Cell Line↗

Isolation of a lipopolysaccharide mutant of Neisseria gonorrhoeae: an analysis of the antigenic and biologic difference.

Analysis of the surface constituents of a pyocin 611 131-resistant variant of strain no. JW-31 of Neisseria gonorrhoeae revealed substantial differences in the lipopolysaccharide (LPS) but not changes in the auxotype or outer-membrane proteins. Immunodiffusion and an enzyme-linked immunosorbent assay showed that the variant strain (no. JW-31R) lost both the LPS serotype and the variable antigens while retaining at least a portion of the common determinant. The use of monoclonal antibody indicated that LPSs from strain no. JW-31R and pyocin 611 131-resistant strains of other LPS serotypes lack a D-galactosaminyl-D-galactopyranosyl-D-glucose moiety. The LPS-derived polysaccharide from strain no. JW-31 binds to wheat-germ lectin in precipitin and inhibition systems, whereas the JW-31R polysaccharide exhibits a markedly reduced affinity. In the presence of normal human serum, 99% of strain no. JW-31R was killed within 20 min and strain no. JW-31 was not.

Animals↗

Studies on the specificity of human IgE-antibodies to the plant proteases papain and bromelain.

The sera of seven patients clinically hypersensitive to papain--in one case also to baromelain--and the sera of sixty asthmatic patients with allergies to other inhalant and food allergens were investigated for IgE antibody activity to the plant proteases papain and bromelain and to common allergens by RAST, confirmed in some sera by RAST inhibition. There seems to be a relation between the antibody reactions to papain and bromelain, in several cases also between the reactions to these proteases and to grass pollen and flour. Studies by RAST inhibition showed that papain, bromelain, wheat flour, rye flour, grass pollen and birch pollen mutually inhibit IgE antibody to each antigen; but the degree of inhibition varies among the different sera and allergens. Our results suggest that these allergens from various plants, besides having specific antigenic determinants, also possess similar or even identical antigenically active regions, leading to immunological cross-reactivity.

Antibody Specificity↗