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Biomedical subjects

D F Smith

Publications and source records attributed to D F Smith.

At least 235 records · Page 13Linked to original sources

Glycolipid-lectin interactions: reactivity of lectins from Helix pomatia, Wisteria floribunda, and Dolichos biflorus with glycolipids containing N-acetylgalactosamine.

The autoradiographic detection of 125I-labeled lectins binding to glycolipids on thin-layer chromatograms can be used to rapidly analyze total glycolipid extracts of cells or tissues for specific oligosaccharide structures. The Helix pomatia lectin which binds with high affinity to terminal alpha-linked GalNAc residues did not bind to globoside (terminal beta 1-3GalNAc) but did bind the ganglioside GM2 and its asialo derivative which have terminal beta 1-4GalNAc residues. The lectin from Dolichos biflorus bound specifically to the Forssman glycolipid with relatively low affinity. The lectin from Wisteria floribunda was bound to Forssman glycolipid, globoside, and the asialo derivative of the ganglioside GM2. The interactions of these lectins with the glycolipid-derived, 3H-labeled oligosaccharides was also analyzed by affinity chromatography. The results indicated that the reactivity of multivalent carbohydrate-binding proteins with polyvalent surfaces of glycolipids is strong enough to permit detection of low-affinity interactions that may not be observed in binding assays that are based on carbohydrate-protein interactions in solution. The autoradiographic analysis of 125I-Helix pomatia lectin binding to thin-layer chromatograms of total lipid extracts from human erythrocyte membranes detected the quantitative differences in the A-active glycolipids from type A1 and A2 cells.

Acetylgalactosamine↗

Studies with antibodies against the conserved cysteine region of progesterone receptor.

Polyclonal antibodies were generated against two synthetic peptides corresponding to sequences from the DNA-binding domain of steroid receptors. The sequence for peptide 1 (13 amino acids) lies between the two putative metal-binding loops of the conserved cysteine region while the sequence for peptide 2 (12 amino acids) lies within one loop. Peptide antibodies were generated by injecting rabbits with peptide conjugated to bovine serum albumin. By Western blot analysis, antibodies to peptide 2 recognized chick and human progesterone receptor and human glucocorticoid receptor, but peptide 1 antibodies did not. No cross-reactivity with native chick progesterone receptor was detected with either anti-peptide. These findings suggest that the epitopes for peptide 2 antibodies, and possibly for peptide 1 antibodies, are inaccessible to antibody in the native receptor.

Amino Acid Sequence↗

Natural infections of phlebotomine sandflies with Trypanosomatidae in central and south Italy.

In a survey of Leishmania infections of phlebotomine sandflies caught in central and south Italy in 1986, zymodeme MON-1 (Montpellier 1) of Leishmania infantum was isolated from Phlebotomus perniciosus and L. tarentolae [= Trypanosoma platydactyli?] was isolated from Sergentomyia minuta, providing for mainland Italy the first direct proof of these long-suspected parasite-vector associations. The roles of P. perfiliewi and P. perniciosus in the transmission of Leishmania spp. in Italy are discussed.

Animals↗

DNA hybridizations on squash-blotted sandflies to identify both Phlebotomus papatasi and infecting Leishmania major.

Epidemiological field studies on leishmaniasis have been hampered by the laborious, and often inefficient, methods used to assess the rates of infection of sandfly vectors (Diptera; Phlebotominae) by species of the causative disease organisms, protozoal parasites of the genus Leishmania (Kinetoplastida; Trypanosomatidae). We report the rapid and accurate identification of both sandfly vector (Phlebotomus (Phlebotomus) papatasi (Scopoli] and infecting Leishmania major Yakimov & Schokov by DNA hybridizations to squash-blotted sandflies. Large numbers of whole (infected) sandflies can be quickly squashed on to nylon hybridization filters and (following standard procedures) the filter-bound DNA can be hybridized sequentially to cloned, multicopy genomic sequences that are specific for species of Leishmania (kinetoplast DNA) or for the sandfly (ribosomal (r) DNA). Our sandfly probe consists of a 3.2 kb fragment of the intergenic 'non-transcribed' spacer of rDNA of P. papatasi that we have detected only in this species: it is present in all six geographically isolated populations tested (from Tunisia through to India) but cannot be detected in the morphologically similar P. (Phlebotomus) duboscqi Neveu-Lemaire, the vector of Leishmania major south of the Sahara; it also cannot be detected in Phlebotomus species of the subgenera Larroussius and Paraphlebotomus that together with P. papatasi are the dominant man-biting sandflies in north African foci of zoonotic cutaneous leishmaniasis, where (as in many arid regions of western Asia) P. papatasi is believed to be the sole vector of L. major.

Animals↗

A comparison of enterotomies through the antimesenteric band and the sacculation of the small (descending) colon of ponies.

Three pairs of longitudinal enterotomies were performed in the small colon of 10 ponies. Each pair consisted of one enterotomy through the antimesenteric band and one through the sacculation. The ponies were destroyed 96 h after surgery. The enterotomies in the two sites were compared for: speed of surgery, adhesions, bursting wall tension, neovascularisation by micro-angiography, and histological reaction. Enterotomies made through the antimesenteric band were superior because they were quicker to perform, stronger, more accurately apposed, and had less inflammation than those made through the sacculation.

Angiography↗

The production of antibodies against the conserved cysteine region of steroid receptors and their use in characterizing the avian progesterone receptor.

The most highly conserved feature of steroid receptor primary structure, the conserved cysteine region, corresponds to the DNA-binding domain of steroid receptors. This domain of chick progesterone receptor (PR) and other receptors was immunochemically characterized using site-directed antibodies. Eight peptides were synthesized corresponding to portion of the conserved cysteine region from PR, glucocorticoid, or estrogen receptor proteins. Polyclonal antibodies were obtained by immunizing rabbits with peptide conjugated to a protein carrier. The cross-reactivity of each antiserum was tested by Western blotting against chick and human PR, human glucocorticoid receptor, and human estrogen receptor. Of the several antisera positive by Western blots, only one cross-reacted with native chick PR. It was found that antibody bound to 4S transformed receptor, but not to 8S nontransformed receptor. At 50 mM KCl, antibody bound preferentially to transformed receptor form A, but at 400 mM KCl antibody bound equally well to either receptor form A or B. Binding of PR to DNA-cellulose was partially inhibited by the presence of cross-reacting peptide antibody. Thus, we have obtained at least one site-directed antibody against steroid receptors which is a useful structural probe to the DNA-binding functional domain of these gene regulatory proteins.

Amino Acid Sequence↗

Sequential presentation of a case of hyperthyroidism with autonomously functioning nodules and Graves' disease in the presence of IgG thyroid stimulators.

The rare occurrence of hyperthyroidism with an autonomously functioning nodule which following 131I therapy presented as toxic diffuse goitre (Graves' disease) is described in a 60 year old male. This progression was characterised by the presence of varying concentrations of IgG thyroid stimulators, thyroid stimulating immunoglobulins and thyroid growth stimulating immunoglobulins, as measured by cytochemical bioassay. It is postulated that the presence of the nodule and its associated hypersecretion of thyroid hormones may have protected the gland from the effects of IgG stimulators by bringing about inhibitory short-loop feedback on normal thyroid cells. It is further suggested that following therapeutic ablation of the nodule, normal thyroid cells became sensitive to the thyroid stimulators with the evolution of typical features of toxic diffuse goitre.

Goiter↗

MR imaging of the cavernous sinus: value of spin echo and gradient recalled echo images.

A detailed evaluation of the MR appearance of the pituitary gland-cavernous sinus junction has not been described. In a series of coronal T1-weighted spin echo images without and with IV gadolinium, we noted the variable size and signal intensity of cavernous venous spaces adjacent to the pituitary gland and the inconsistent visualization of the dural membrane just lateral to the gland. Correlation of coronal T1-weighted spin echo and gradient recalled echo images (the latter with high-signal-intensity vascular structures) proved to be an effective means of identifying cavernous venous spaces, connective tissue and cranial nerves, and the lateral margins of the pituitary gland, and of differentiating tumor tissue from cavernous venous spaces. Further work is needed to develop criteria to distinguish cavernous sinus compression from actual tumor invasion.

Adenoma↗

Comparative healing of mesenteric and antimesenteric incisions in the bovine jejunum.

Paired incisions (n = 6 pairs) were made in the jejunum of each of 8 Holstein cows. Pairs consisted of 2-cm transverse incisions, 35 cm apart, at the antimesenteric region and at the mesenteric region. Bursting-wall tension was used to test mechanical strength, and cellular and vascular events of healing were evaluated using histologic examination and microangiography. Healing was evaluated at postoperative hour (POH) 48 in 4 of the cows (group 1) and at POH 96 in the remaining 4 (group 2). Evidence of leakage of intestinal contents was not found in any of the cows. At POH 48 and 96, bursting-wall tension was significantly (P less than 0.001) greater in the intestinal segments with antimesenteric incisions than in those segments with mesenteric incisions. Disruption of normal vasculature was seen at mesenteric and antimesenteric sites; ingrowth of vessels, reformation of vascular plexuses, and development of collateral circulation were observed at POH 96. Granulation tissue was observed at POH 96 at the antimesenteric and mesenteric sites, and early stages of mucosal reepithelialization were seen in several sections at POH 48. Better apposition of tissue layers was seen in antimesenteric incisions, but mucosal eversion was evident in several mesenteric incisions. Some sections examined from cows at POH 96 had complete bridging of mucosa, submucosa, and muscularis layers at the antimesenteric location. Inflammatory cells were observed along the incision and at the mucosal and serosal surfaces in many of the sections. Deposition of new collagen was not appreciable in any section at POH 48 or 96.

Animals↗

Postinfusion phlebitis after intravenous push versus intravenous piggyback administration of antimicrobial agents.

The incidence and severity of phlebitis associated with the i.v. push method versus the i.v. piggyback method of administration of antimicrobial agents were evaluated. All patients admitted to two nursing units of an 886-bed, tertiary-care hospital from March through June 1987 who received intravenous cephalosporins or extended-spectrum penicillins were considered for inclusion in the study. The method of administration, either i.v. push or i.v. piggyback, was randomly assigned. Each patient's catheter site was evaluated every 24 hours according to standardized definitions of phlebitis and patency. Information on each patient and venipuncture site also was collected. A total of 319 catheter sites were studied in 155 patients. There was no significant difference in the incidence or severity of postinfusion phlebitis between the i.v. push and the i.v. piggyback groups (41% versus 47%). When studying only those catheter sites discontinued because of phlebitis, the time until development of phlebitis averaged 45 +/- 20.5 hours in the i.v. push group, compared with only 36.2 +/- 17.6 hours in the i.v. piggyback group; this difference was significant. The fact that the catheter sites lasted significantly longer in the i.v. push group, combined with elimination of the cost of syringe infusion pumps or i.v. tubing and minibags, suggests that use of the i.v. push method may result in substantial cost savings. Other advantages associated with the i.v. push method include time savings for nursing personnel and patients and greater ease in scheduling doses of multiple antimicrobial agents.(ABSTRACT TRUNCATED AT 250 WORDS)

Aged↗

Expo '86, Vancouver: impact on British Columbia's Children's Hospital.

Over 22 million visitors attended the 1986 world exposition in Vancouver, and this had a significant impact on the local pediatric facility. A total of 559 children visited the emergency department of British Columbia's Children's Hospital with injuries or illnesses resulting from the fair. Of these, 193 (34%) had come directly from the site. The children's ages ranged from 1 1/2 months to 18 years, 4 months (mean 6.99 years). Of the 559 children 31% were not covered by medical insurance. Twenty-four of the children were admitted to the hospital, and one died at the Expo site. Most of the illnesses were upper respiratory tract infections and gastroenteritis. The proportion of visits for trauma was 50%, compared with the yearly average of 25%. There was a 6% increase in the workload in the emergency department during the fair; the percentage was even higher during July and August, particularly after 1800 hours. Recommendations are made to aid with the planning of medical care for similar events.

Adolescent↗

III6NeuAcLc4Cer in human SW1116 colorectal carcinoma cells: a possible oncofetal antigen that is not dependent on Lewis gene expression.

Monospecific rabbit antibodies directed against the human milk sialyloligosaccharides III6NeuAcLcOse4 (sialyltetrasaccharide b), IV3NeuAcLcOse4 (sialyltetrasaccharide a), and IV6NeuAcnLc4Ose (sialyltetrasaccharide c) were used to detect their homologous haptens as gangliosides or ganglioside-derived sialyloligosaccharides from the human colorectal carcinoma cell line SW1116. III6NeuAcLc4Cer was first detected in human meconium [P. A. Prieto and D. F. Smith (1985) Arch. Biochem. Biophys. 241, 281-289], and its presence in a total ganglioside fraction of SW1116 cells together with its absence from a total lipid extract of normal human intestinal mucosa are consistent with III6NeuAcLc4Cer being a tumor-associated oncofetal antigen. IV3NeuAcLc4Cer, a ganglioside in human meconium [P. A. Prieto and D. F. Smith (1986) Arch. Biochem. Biophys. 249, 243-253], was also detected in SW1116 cells; an observation that is consistent with its being the immediate precursor to the sialyl-Lea ganglioside in SW1116 cells. Specific antisera against sialylated type 1 oligosaccharide chains whose expression is independent of the Lewis gene fucosyltransferase may be useful diagnostic reagents for oncofetal, carbohydrate antigens.

Antibodies↗

A novel sialylfucopentaose in human milk. Presence of this oligosaccharide is not dependent on expression of the secretor or Lewis fucosyltransferases.

We have identified a novel oligosaccharide in human milk that is a fucosyl derivative of sialyltetrasaccharide c (NeuAc alpha 2-6Gal beta 1-4GlcNAc beta 1-3Gal beta 1-4Glc). This oligosaccharide was purified by affinity chromatography on a column of immobilized Ricinus communis I lectin. Structural analyses of radiolabeled oligosaccharides by exoglycosidase digestions, binding by specific anti-carbohydrate antibodies, and analysis of the 3H-labeled glucitol derivative obtained after permethylation and hydrolysis are consistent with the following proposed structure. (formula; see text) The analyses of human milk sialylpentasaccharides from donors typed as Le(a-,b+), Le(a+,b-), and Le(a-,b-) secretor confirmed the secretor gene-dependent expression of the sialylated lacto-N-fucopentaose I (Fuc alpha 1-2Gal beta 1-3[NeuAc alpha 2-6]GlcNAc beta 1-3Gal beta 1-4Glc) and the Lewis gene-dependent expression of the sialylated lacto-N-fucopentaose II (NeuAc alpha 2-3Gal beta 1-3[Fuc alpha 1-4]GlcNAc beta 1-4Glc). However, the presence of this novel oligosaccharide in human milk is not dependent on the expression of either the secretor gene or the Lewis gene-specified fucosyltransferases.

Carbohydrate Sequence↗

Toxin A from Clostridium difficile binds to rabbit erythrocyte glycolipids with terminal Gal alpha 1-3Gal beta 1-4GlcNAc sequences.

The binding of Toxin A isolated from Clostridium difficile to rabbit erythrocyte glycolipids has been studied. Total lipid extracts from rabbit erythrocytes were subjected to thin-layer chromatography and toxin-binding glycolipids detected by using 125I-labeled Toxin A in a direct binding overlay technique. Two major and several minor toxin-binding glycolipids were detected in rabbit erythrocytes by this method. The results of structural analyses of the major toxin-binding glycolipids were consistent with a pentasaccharide-ceramide (Gal alpha 1-3Gal beta 1-4GlcNAc beta 1-3Gal beta 1-4Glc-Cer) and a branched decasaccharide-ceramide (Gal alpha 1-3Gal beta 1-4GlcNAc beta 1-3[Gal alpha 1-3Gal beta 1-4GlcNAc beta 1-6]Gal beta 1-4GlcNAc beta 1-3Gal beta 1-4Glc-Cer) previously identified as the two most abundant glycolipids in rabbit erythrocytes. 125I-Toxin A binding to these glycolipids could be inhibited by bovine thyroglobulin, monospecific antiserum to the toxin, or by treatment of the glycolipids with alpha-galactosidase. The absence of toxin interaction with isoglobotriaosylceramide (Gal alpha 1-3Gal beta 1-4Glc-Cer) isolated from canine intestine suggested that the GlcNAc residue present in the terminal Gal alpha 1-3Gal beta 1-4GLcNAc sequence common to all known toxin binding glycoconjugates is required for carbohydrate-specific recognition by Toxin A. These observations are consistent with the proposed carbohydrate binding specificity of Toxin A for the nonreducing terminal sequence, Gal alpha 1-3Gal beta 1-4GlcNAc.

Animals↗

Thrombosis of upper extremity thoracic inlet veins: diagnosis with duplex Doppler sonography.

Duplex Doppler sonography of the axillary, subclavian, internal jugular, and brachiocephalic veins was performed in 20 normal volunteers and in a series of 22 consecutive patients with suspected venous thrombosis. When normal, these vessels had hypoechoic or anechoic lumina, sharply echogenic walls, and characteristic Doppler flow patterns that varied with changes in intrathoracic pressure. Most thrombosed vessels had poorly defined walls and echogenic lumina. All had absent flow on Doppler analysis. Duplex Doppler sonography was positive in all 10 patients subsequently shown to have venous thrombosis by venography or CT. One false-positive result occurred in a technically limited study. Sonography has the advantages of portability, ease of performance, and high patient acceptance. Duplex Doppler has advantages over either continuous-wave Doppler or imaging sonography alone. Our experience suggests that duplex Doppler sonography should be the initial diagnostic study in evaluating suspected thrombosis of the upper extremity thoracic inlet veins.

Adolescent↗