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Biomedical subjects

G D Cain

Publications and source records attributed to G D Cain.

At least 19 recordsLinked to original sources
PubMed1996

Molecular characterization of Echinostoma caproni and E. paraensei by random amplification of polymorphic DNA (RAPD) analysis.

Genomic DNA from Echinostoma caproni and Echinostoma paraensei was amplified by the polymerase chain reaction using primers (5'-TCGTAGCCAA and 5'-TCACGATGCA), originally found to differentiate species and strain of Schistosoma. The 2 putative species of Echinostoma produced distinct banding patterns clearly distinguishable from one another, thereby suggesting that RAPD (random amplification of polymorphic DNA) analysis may be useful for the identification of echinostome strains and species previously misunderstood or undescribed, and that primers developed for species within a given genus, e.g., Schistosoma, may have broader application in identifying other trematodes.

Animals↗
PubMed1991

[Comparative studies on the repetitive DNA sequences of diploid and triploid forms of Paragonimus westermani by restriction endonuclease and Southern blotting].

Restriction enzyme digestion of total genomic DNA of two chromosomes forms of Paragonimus westermani showed the presence of homologous highly repeated DNA in both diploid and triploid forms. Southern blotting analysis provided further evidence that the distribution of restriction enzyme sites (with 3 enzymes) on repetitive sequence of DNA of both forms were similar. However, with Pstl, Ddel, HaeIII and HpaII, their polymorphism revealed differences which were also found in each form tested separately with the hybridization technique. The present study, at the molecular level, supports the previously reported biological and biochemical results that they might be considered as different isolates or forms. It is suggested that PstI, DdeI, HaeIII and HpaII digestion pattern might be useful to distinguish the diploid from the triploid forms of Paragonimus westermani.

Animals↗
PubMed1990

The gene family encoding eggshell proteins of Schistosoma japonicum.

The four closely related genes encoding eggshell proteins in the human parasite Schistosoma japonicum are described. A cDNA and a genomic DNA library were constructed and members of the eggshell protein gene family isolated. The four genes in this family do not contain introns, and differ in organization and nucleotide sequence from the related set of genes in Schistosoma mansoni and Schistosoma haematobium. The coding sequences of two of the S. japonicum genes and their flanking regions were determined. Transcription start sites for these genes were shown by primer extension analysis to occur 47 and 50 nucleotides in front of the start codon. A female-specific component in nuclear extracts binds to a DNA fragment containing conserved sequences upstream of the transcription start sites. The deduced protein sequences of 207 and 212 amino acids are composed of 50% glycine with continuous glycine regions as long as 11 residues. In vitro translations of male and female RNAs revealed female-specific translation products, the sizes of which were consistent with the eggshell proteins.

Amino Acid Sequence↗
PubMed1989

Soluble proteins from Schistosoma mansoni and japonicum: a comparative biochemical and immunological analysis.

1. Soluble proteins were recovered from male Schistosoma mansoni after homogenization in Tris-HCl buffer containing 0.6 M KCl and 1.0% Triton X-100 followed by preparative electrophoresis on SDS-gel. 2. Polyclonal antibodies produced in mice against the soluble fraction were used in comparative analysis of S. mansoni and S. japonicum using immunoblots and immunoprecipitation of in vitro translated polypeptides. 3. Small molecular weight polypeptide (20-22 kdalton), identified by infected mouse serum (IMS) on immunoblots, was predominant in females and was not cross-reactive with heterologous IMS. 4. A 41-43 kdalton polypeptide which appeared as a doublet on immunoblots performed with polyclonal antiserum 4M, was predominant in males of both species although the polypeptides of S. mansoni showed slower electrophoretic mobility, and therefore the larger size (43 kdalton), than that of S. japonicum. 5. Comparison of fluorograms of the immunoprecipitates of in vitro translated polypeptides indicated that IMS of S. mansoni precipitated two, 30 and 94 kdalton, polypeptides while the IMS of S. japonicum identified at 72 kdalton polypeptide. Antisera 1M, 2M and 4M also showed similarities and differences in polypeptides of in vitro translation products of the two species of Schistosoma.

Animals↗
PubMed1988

The selective uptake of cholesterol by the rat tapeworm Hymenolepis diminuta (Cestoda).

1. The sterols of Hymenolepis diminuta are almost exclusively cholesterol or similar C-27 sterols; the free sterols of its environment (the lumen of the rat intestine) are cholesterol and various phytosterols. 2. During incubation of tapeworms with mixed micelles of taurocholate, glyceryl monooleate, and equimolar [3H]cholesterol and [14C]beta-sitosterol, the uptake of cholesterol is 40 times more rapid than the uptake of sitosterol. 3. Following uptake, the desorption of labeled sitosterol is six times more rapid than that of cholesterol. 4. We did not detect the esterification of absorbed sterols or the conversion of absorbed sitosterol of cholesterol. 5. The highly selective uptake of cholesterol and the moderately selective desorption of phytosterols can account for the selective accumulation of C-27 sterol by the tapeworm.

Animals↗
PubMed1988

Studies on the mechanism of cholesterol uptake by the rat tapeworm Hymenolepis diminuta (Cestoda).

1. With increasing cholesterol content in mixed micelles, the rate of cholesterol uptake by the tapeworm approaches a limiting, maximal value. 2. This uptake is inhibited only 32-40% by other sterols, but is not markedly dependent on medium pH or tapeworm energy metabolism. 3. The competitive exchange diffusion of absorbed [14C]cholesterol could not be demonstrated. 4. The above results partially support the hypothesis that the tapeworm absorbs cholesterol by a specific carrier-mediated process. 5. Cholesterol uptake is reduced when the capacity of the micellar phase of the medium is increased, suggesting that uptake involves the intermediate partitioning of sterol from micelles into the aqueous phase of the medium.

Absorption↗
PubMed1987

Actin and intermediate-sized filaments of the spines and cytoskeleton of Schistosoma mansoni.

The organization of spines and filaments in whole worms and cytoskeletal fractions of adult Schistosoma mansoni was investigated. The ultrastructure of the spine revealed a closely packed filamentous organization of 3.5- to 5.6-nm elements and electron-lucent areas. Spines were surrounded at the base by electron-dense bodies and membrane invaginations, and covered at the tip by the syncytial surface membrane. Filaments, 7.5-11.1 nm in diameter, were closely associated with the base of the spines, between muscles, near mitochondria or nuclei, and in spaces of the subtegument. Cytoskeletal fractions prepared by homogenizing adults in Tris-HCl buffer, containing 0.6 M KCl and 1.0% Triton X-100, represented 19%-25% and 32%-38% of wet weight of males and females, respectively. The fractions contained nuclei, spines, 8 to 11 nm filaments, myofibrils, and granules. Vitellaria and egg shells were abundant in fractions from females. Six polypeptides with estimated molecular weights of 130, 96, 84, 78, 74 and 43 kdaltons were identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis as the major components of the cytoskeleton. Monoclonal antibody to chicken actin (MAA) was localized predominantly in surface spines and tubercles of adult schistosomes by the indirect immunofluorescence test, while immune serum from infected mice reacted less specifically with the tegument. A 43-kdalton polypeptide with electrophoretic mobility identical to that of vertebrate actin, identified in cytoskeletal and tegumental fractions of adult worms, reacted positively with MAA on immunoblotting.(ABSTRACT TRUNCATED AT 250 WORDS)

Actins↗
PubMed1985

tris(Hydroxymethyl)aminomethane: an artifact detected as an aminotetrose by gas chromatography/mass spectrometry.

The monosaccharide content of the ethanol-soluble moiety of crude polysaccharide fractions of the tapeworm Hymenolepis diminuta and the blood fluke Schistosoma mansoni was investigated by gas-liquid chromatography of alditol acetate derivatives. A prominent, unusual peak was found, and mass spectrometry suggested identification as an aminotetrose, a four-carbon aminosugar reported only once in the past 15 years [D. A. Cumming, C. G. Hellerquist, and O. Touster (1981) J. Biol. Chem. 256, 7723-7726]. The unknown molecule was found to be tris(hydroxymethyl)aminomethane (Tris), whose acetylated derivative has a mass spectrum identical to that of an aminotetrose. The source of the Tris was traced to buffers used in conjunction with polysaccharide isolation.

Amino Sugars↗
PubMed1985

Biosynthesis of polyisoprenoid lipids in the rat tapeworm Hymenolepis diminuta.

The possible occurrence of isoprenoid lipids in the tapeworm, Hymenolepis diminuta, was investigated by analytic and biosynthetic methods. Two-dimensional thin layer chromatography (TLC) resolved the worm's non-saponifiable lipids into cholesterol, farnesol, and several unknown compounds, two of which migrated with dolichol standards on TLC and reacted with phthalic anhydride, a probe for alcohols; the major compound also exhibited a mass spectrum very similar to that of a dolichol standard. A third unknown compound separable by TLC, apparently a quinone, was intrinsically red, was decolorized by treatment with sodium dithionite and migrated on TLC in a more polar position than either ubiquinone-50 or vitamin K1. All three compounds, as well as farnesol, were labelled when worms were incubated with [14C]-mevalonolactone, suggesting that they are endogenous isoprenoids.

Animals↗
PubMed1985

Isolation and characterization of glycosaminoglycans from Schistosoma mansoni.

Tegumental tissues of paired adult Schistosoma mansoni were removed by treatment with Triton X-100 and recovered by centrifugation. The chloroform-methanol insoluble residues of this isolated tegumental fraction and of the denuded carcasses were analysed for glycosaminoglycan (GAG) and sialic acid contents. Treatment with GAG-specific enzymes followed by electrophoretic analysis showed that both the carcass and tegument contained heparin and/or heparan sulfate, chondroitin sulfate and hyaluronic acid. All these except hyaluronic acid were present in the tegumental fraction. Based on uronic acid content, about 73% of the total GAG was in the tegumental membrane, 15% in the tegmental matrix and the remaining 12% was in the carcass. The presence of heparin-like polysaccharide may present entrapment of the schistosoma by the hosts' blood-clotting process.

Animals↗
PubMed1985

A comparison of Texan and Hawaiian strains of the avian eyefluke, philophthalmus gralli, with a cautionary note on the importation of exotic animals.

Philophthalmus gralli (Mathis and Leger, 1910) was introduced into the San Antonio, Texas area within the last 25-30 years from an unknown foreign source. Strains of P. gralli originally from Texas and Hawaii were compared for differences in morphology, growth patterns, mating compatibility, and isozyme mobilities. Metacercarial cysts of the 2 strains were compared for viability after storage at room temperature. Adult stages of the 2 strains were indistinguishable based on sucker ratios, vitellaria, and egg sizes. In single- and multiple-worm infections, both strains exhibited similar growth patterns when reared in chickens. Adults of the 2 strains, when transplanted in concurrent infections, readily cross-inseminated. Isozyme mobilities of 5 enzymes were identical in all adult worms examined from both strains. After 3 days, metacercarial cysts from both strains rapidly lost their ability to excyst. The cysts of the Texan strain remained viable for 4 days longer than the Hawaiian strain, although excystment rates were low during that period. The evolutionary implications of this imported species are discussed.

Animals↗
PubMed1984

Amino acid and lipid composition of refringent granules from the ameboid sperm of Ascaris suum (Nematoda).

Transformation of the spermatozoon of Ascaris suum from a spheroidal to an ameboid cell is associated with the formation of a motile pseudopodium and coalescence of the intracellular refringent granules. The pseudopodia of the ameboid spermatozoa contain filaments organized into dense patches, bundles, web-like or lace-like networks, as observed by electron microscopy. The morphology and chemistry of the refringent granules were investigated in subcellular fractions enriched for these structures. Isolated refringent granules were heterogeneous in size measuring from 0.5 X 0.6 to 2.3 X 3.5 microns. Each granule is surrounded by a 110 A thick layer. During fusion, the surfaces of the refringent granules form small extensions resembling micropodia. The process of fusion occurs at many sites on a given granule and simultaneous fusion of several granules was commonly observed. Amino acid analyses of the refringent granule proteins (RGP's) indicated: they are rich in aspartic acid or asparagine (48%), leucine (10%), serine (19%) and aromatic amino acids (11%). Gas-liquid chromatographic analyses of alditol acetate derivatives of monosaccharides released by mild acid hydrolysis showed the predominant sugars to be glucose (7.3 micrograms/mg protein), galactose (9.2 micrograms/mg) and N-acetylglucosamine (5.5 micrograms/mg). Lipid analyses indicated a complex mixture of glycerides, ascarosides and waxes, together with a major component that resembled free fatty acid in mobility on TLC.

Amino Acids↗
PubMed1984

Characterization of polysaccharides of the eggs and adults of Hymenolepis diminuta.

Polysaccharides and other complex carbohydrates were released by proteolysis of the chloroform-methanol insoluble residue of 10 day-old worms and eggs of Hymenolepis diminuta. Gas-liquid chromatographic analysis of alditol acetate derivatives of monosaccharides released from the polysaccharides by hydrolysis revealed that in the 10 day-old worm, glucose was the most abundant sugar, followed by galactose, glucosamine, galactosamine, fucose and possibly rhamnose. Mannose was least abundant and xylose was absent. In the egg, glucose and galactose were equally abundant, followed by the same sugars found in 10 day-old worms, and xylose was present. Uronic acid was detected in both fractions by specific chemical tests. None of the saccharide material from eggs and worms was susceptible to degradation by Streptomyces hyaluronidase, chondroitinase AC, and slightly susceptible to chondroitinase ABC, as shown by electrophoretic analysis on composite 2.2% acrylamide-agarose slab gels and 4.5/12.5% polyacrylamide gels before and after enzymatic treatment. One of the gel-separable bands, however, was degradable by both nitrous acid and Flavobacterium heparinase. Both bands from eggs were degradable by nitrous acid. These results suggest that eggs contain heparin and/or heparan sulfate and perhaps dermatan sulfate and that 10 day-old worms also have these polyglycans but possibly not chondroitin sulfate or hyaluronic acid.

Amino Sugars↗
PubMed1984

Glycosaminoglycans of tegumental fractions of Hymenolepis diminuta.

The teguments of 6 and 10 day-old Hymenolepis diminuta were removed with Triton X-100 and separated into brush border and vesicular fractions by differential centrifugation. Glycosaminoglycans (GAG) isolated from these tissues and from the denuded carcass were treated with specific GAG-degrading enzymes and other chemical agents and analyzed by sodium dodecyl sulfate-polyacrylamide, agarose gel and cellulose acetate electrophoresis. Both 6 and 10 day-old worm carcasses contained chondroitin sulfate, heparin/heparan sulfate and hyaluronic acid. The 10 day-old worm brush border and vesicle fractions contained chondroitin sulfate but no heparin-like material. Colorimetric analysis showed that the carcasses of both 6 and 10 day-old worms contained uronic acid. About 98% of the detectable uronic acid of 10 day-old worms was found in the carcass, and only 2% in the brush border fraction. No uronic acid was detected in the other tegumental fractions.

Aging↗
PubMed1983

Comparison of the expression-linked extra copy (ELC) and basic copy (BC) genes of a trypanosome surface antigen.

A recombinant clone of an expression-linked extra copy (ELC) gene of a trypanosome-variable surface glycoprotein was sequenced. In addition the sequences of the corresponding cDNA and portions of the two basic copy genes were determined. Comparison of these sequences reveals that the 5' boundary of the ELC-transposed segment (2.2 kb) occurs within a repetitive sequence about 700 bp upstream from the start codon of the coding sequence. This sequence does not contain internal symmetries and is not homologous with the repetitive sequence at the 3' boundary. The first 35 nucleotides of the cDNA are different than the corresponding ELC sequence and presumably were transcribed from another genomic location. A restriction fragment containing predominantly sequences outside of the 5' boundary hybridizes to a Pst I fragment whose length is variable in different trypanosome clones. This hybridization pattern is similar to that observed using probes for surface glycoprotein genes that are expressed via the nonduplication-associated (NDA) mechanism rather than the ELC mechanism. This indicates that there is a sequence correlation between these two DNA rearrangement mechanism.

Amino Acid Sequence↗
PubMed1983

Comparison of calcium, freeze-thaw, and Triton X-100 tegumental disruption/recovery techniques applied to Schistosoma mansoni.

Three techniques for the disruption/recovery of tegumental free-surface plasmalemma were compared by (i) morphological examination of carcasses and centrifugally-derived isolates, (ii) specific enrichment of bound surface tags (lectin) and of "marker" enzymes for membrane, and (iii) assessment of total protein and lectin recovered by each procedure. Procedures compared included the use of Triton X-100, freezing and thawing, and high ionic strength calcium. Triton X-100 consistently provided the greatest amounts of recovered surface membrane on a per worm basis, whereas calcium retained the highest amounts of alkaline p-nitrophenyl phosphatase, adenosine triphosphatase, and adenosine monophosphatase activity. Ultrastructural examination of membrane isolates and worm carcasses prepared by freezing and thawing indicated that significant amounts of parenchymal material contaminated the membrane fractions. Thus results based on the freeze-thaw technique can be difficult to interpret.

5'-Nucleotidase↗
PubMed1983

Recurrent ulcers after gastric surgery: endoscopic localization to the gastric mucosa.

The traditional teaching has been that ulcers recurring after ulcer surgery occur at, or distal to, the anastomosis of the gastric and intestinal mucosa. These concepts predate the widespread availability of fiberoptic endoscopy. We noted that recurrent ulcers were frequently located in the gastric mucosa proximal to the gastrointestinal anastomosis. We analyzed our cases for the purpose of identifying the location of ulcers that occur after surgery for peptic ulcer disease and to investigate the possible relationship between indication for the original ulcer surgery and the presentation of the recurrent ulcer. Ulcers after peptic ulcer surgery were most frequently located in the gastric mucosa proximal to the intestinal anastomosis. The location of the recurrence was not found to be related to the original diagnosis, to the indication for the operation, or to the presentation of the recurrence. These findings may reflect either a change in the pattern of ulcer recurrence or an altered pattern of recognition related to the routine use of endoscopy.

Adult↗