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Biomedical subjects

L A Ellis

Publications and source records attributed to L A Ellis.

At least 19 recordsLinked to original sources
PubMed2001

Genetic analysis of the connexin-26 M34T variant: identification of genotype M34T/M34T segregating with mild-moderate non-syndromic sensorineural hearing loss.

Mutations in the human gap junction beta-2 gene (GJB2) that encodes connexin-26 have been shown to cause non-syndromic sensorineural hearing loss (NSSNHL) at the DFNB1 locus on 13q11. Functional and genetic data regarding the disease causing potential of one particular GJB2 sequence variant, 101 T-->C (M34T), have proven contradictory. In this study, we found the prevalence of the M34T allele in a cohort of white sib pairs and sporadic cases with NSSNHL from the United Kingdom and Ireland to be 3.179% of chromosomes screened. Significantly, we identified the first M34T/M34T genotype cosegregating in a single family with mid to high frequency NSSNHL. Screening a control population of 630 subjects we identified 25 M34T heterozygotes; however, no M34T homozygotes were detected. Surprisingly, the majority of M34T alleles (88%) were in cis with a 10 bp deletion in the 5' non-coding sequence. This non-coding deletion was also homozygous in the homozygous M34T subjects. Microsatellite analysis of flanking loci in M34T heterozygotes and controls does not define an extensive ancestral haplotype but preliminary data suggest two common alleles in subjects with the M34T allele. In summary, we provide data that support M34T acting as a recessive GJB2 allele associated with mild-moderate prelingual hearing impairment.

Alleles↗
PubMed2001

Increased chromosome translocations and aneuploidy in peripheral blood lymphocytes of patients having revision arthroplasty of the hip.

The long-term biological effects of wear debris are unknown. We have investigated whether there is any evidence of cumulative mutagenic damage in peripheral blood lymphocytes of patients undergoing revision arthroplasty of predominantly metal-on-plastic total hip replacements compared with those at primary arthroplasty. There was a threefold increase in aneuploidy and a twofold increase in chromosomal translocations which could not be explained by the confounding variables of smoking, gender, age and diagnostic radiographs. In the patients with TiVaAl prostheses there was a fivefold increase in aneuploidy but no increase in chromosomal translocations. By contrast, in patients with cobalt-chrome prostheses there was a 2.5-fold increase in aneuploidy and a 3.5-fold increase in chromosomal translocations. In six patients with stainless-steel prostheses there was no increase in either aneuploidy or chromosomal translocations. Our results suggest that future epidemiological studies of the putative long-term risks of joint replacement should take into account the type of alloy used in the prosthesis.

Aged↗
PubMed2000

A comparison of fluorescent SSCP and denaturing HPLC for high throughput mutation scanning.

We examined 67 different mutations in 16 different amplicons in a comparison of mutation detection by fluorescent single strand conformation polymorphism (F-SSCP) and by denaturing HPLC (DHPLC). F-SSCP was used to analyze fluorescent amplicons with internal size standards and automated fragment analysis (GeneScan, PE Applied Biosystems, Foster City, CA). In DHPLC, unlabelled amplicons were analyzed by reverse phase HPLC with fragment detection by absorbance at 260nm. Both methods had high sensitivity (95-100%) and specificity (100%). Overall, F-SSCP with external temperature control was the more sensitive method, but DHPLC was particularly useful for the rapid analysis of novel fragments.

Chromatography, High Pressure Liquid↗
PubMed1997

Chromatographic and hyphenated methods for elemental speciation analysis in environmental media.

This review discusses chromatographic techniques that permit the analysis of speciated metals in the environment using conventional detectors, such as UV, and element-specific detectors, such as flame atomic absorption spectrometry, electrothermal atomic absorption spectrometry and inductively coupled plasma mass spectrometry. The importance of determining precise elemental forms in hazardous waste-contaminated soil, water and biota in terms of toxicity is outlined. Previous reviews on this subject are described and recent research on this subject is discussed. Most of the work cited has been performed in the 1990s and a table summarizing the chromatographic method and the detector system used, including brief comments on the work, is included to enable quick reference.

Animals↗
PubMed1997

Terminal beta-linked tyvelose creates unique epitopes in Trichinella spiralis glycan antigens.

Indirect evidence that the immunodominant N-glycans of the parasite, Trichinella spiralis are capped by novel beta-linked 3,6-dideoxy-D-arabinohexopyranosyl residues (tyvelose, Tyv) was obtained from immunochemical assays employing monoclonal antibodies and synthetic oligosaccharides. Three of four previously characterized monoclonal antibodies generated from the lymphocytes of T.spiralis infected rats bind BSA glycoconjugates bearing the synthetic epitope beta-D-Tyvp(1-->3)-beta-D-GalNAcp but not to the corresponding alpha-D-Tyvp(1-->3)-beta-D-GalNAcp-BSA glycoconjugate. Monosaccharide and disaccharide glycoside inhibition data mirrors the results of the direct binding experiments. The branched tetrasaccharide beta-D-Tyv(1-->3)-beta-D-GalNAcp(1-->4)[alpha-L-Fucp(1 -->3)] beta-D-GalNAcp is the most active synthetic oligosaccharide inhibitor for all four monoclonal antibodies studied, while the corresponding alpha-D-Tyv containing tetrasaccharide and the core trisaccharide beta-D-GalNAcp(1-->4)[alpha-L-Fucp(1-->3)]beta-D-GlcNAcp+ ++ are inactive. The exceptional inhibitory activity of the disaccharide beta-D-Tyvp(1-->3)-beta-D-GalNAcp with one mAb (18H) compared to that of the branched tetrasaccharide beta-D-Tyvp(1-->3)-beta-D-GalNAcp(1-->4)[alpha-L-Fucp( 1-->3)]-beta-D-GlcNAcp is indicative of the presence of linear, nonfucosylated glycan epitopes (beta-D-Tyvp(1-->3)-beta-D-GalNAcp(1-->4) beta-D-GlcNAcp) that lack a fucose residue in one arm of the antigenic, tetra-antennary N-glycan. This observation supports earlier FAB-mass spectrometry evidence for the existence of tetra-antennary, core fucosylated glycans that lack a fucose residue on one of their antennae.

Animals↗
PubMed1997

Do milk-borne cytokines and hormones influence neonatal immune cell function?

Cytokines, growth factors and various hormones collectively control the proliferation, survival, differentiation and function of immune cells. A wide array of these compounds is present in maternal milk and ingested by neonates during a period of rapid maturation of gut-associated and peripheral lymphoid tissues. The functional consequences of most milk immunomodulatory constituents in neonates are unknown. However, there is evidence that milk prolactin acts as a developmental regulator of the neonatal immune system, supporting the premise that milk constituents with immunomodulatory activity may serve as neonatal immunodevelopment agents.

Animals↗
PubMed1997

Digestive enzymes in human milk: stability at suboptimal storage temperatures.

BACKGROUND: Women who return to work outside of the home while still breastfeeding must often store the expressed milk at less than optimal temperatures. Human milk provides digestive enzymes (amylase and lipase) that compensate in the newborn for immature pancreatic function. METHODS: We have assessed the stability of amylase and bile salt-dependent lipase after storage for 1-24 h at 15, 25, and 38 degrees C. RESULTS: Both enzymes were stable at 15 and 25 degrees C for 24 h, whereas at 38 degrees C there was a 15 and 20% decrease in lipase and amylase activity, respectively. The stability of milk lipoprotein lipase was also tested. This very labile enzyme was more stable in milk than previously reported for blood and tissues, i.e., 20 and 50% decrease in activity after storage at 15 or 25 degrees C for 24 h, respectively. A two-unit drop in milk pH by 24 h of storage would not affect the activity of digestive enzymes, which are stable at pH > 3.5. CONCLUSIONS: We conclude that milk provides the same compensatory digestive activity after short-term storage, even at relatively high temperature, as when fed fresh to the infant.

Adult↗
PubMed1996

Milk-borne prolactin and neonatal development.

Milk is primarily regarded as a food furnishing essential nutrients for infant growth and development, but milk can also serve as a vehicle for mother to neonate transfer of molecules that regulate development. A wide array of biologically active compounds such as hormones, cytokines and enzymes are present in milk, especially early milk. The premise that prolactin (PRL) in milk is an important and possibly essential developmental factor for the newborn is explored. Both PRL and structurally modified isoforms are abundant in early milk and gradually diminish with the progression of lactation. Milk PRL is absorbed and biologically active in the neonate. Assays of PRL variants, experimental paradigms to test them as developmental regulators and the body of evidence supporting the hypothesis that milk PRL regulates differentiation and maturation of neonatal neuroendocrine, reproductive, and immune systems is presented.

Breast Feeding↗
PubMed1996

Breastfeeding and the working mother: effect of time and temperature of short-term storage on proteolysis, lipolysis, and bacterial growth in milk.

BACKGROUND: Women who breastfeed have to store expressed milk while at work for later feeding to their infants; however, storage conditions are often not optimal. OBJECTIVE: Top assess microbial growth and stability of milk protein and lipid at 15 degrees C to 38 degrees C for up to 24 hours. METHODS: Sixteen healthy women who breastfed exclusively, either at home (n=11) or who expressed milk for their infants (n=5), were studied during early (1 month) or late (5 to 6 months) lactation. Expressed milk was stored at 15 degrees C, 25 degrees C, and 38 degrees C for 1 to 24 hours for quantitation of pH, proteolysis, and lipolysis; bacterial growth was quantified at 0, 4, 8, and 24 hours of storage. RESULTS: Milk pH decreased 2 units by 24 hours of storage at all temperatures tested. Proteolysis was minimal during milk storage at 15 degrees C or at 25 degrees C for 24 hours and was apparent only after 24 hours of storage at 38 degrees C. Lipolysis was rapid, starting in the first hours of storage and progressing to 8% at 24 hours. Thus, while the greatest increment in proteolysis products was a 40% increase above baseline after 24 hours of storage at 38 degrees C, free fatty acid concentration at this storage time was 440% to 710% higher than in freshly expressed milk. Bacterial growth was restricted mainly to nonpathogens, was minimal at 15 degrees C throughout the 24 hours of storage, was low at 25 degrees C for the first 4 to 8 hours, and was considerably higher at 38 degrees C even during the relatively short period of 4 hours. CONCLUSIONS: Storage of human milk is safe at 15 degrees C for 24 hours, whereas at 25 degrees C it is safe for 4 hours. Milk should not be stored at 38 degrees C. Minimal proteolysis during storage suggests that milk proteins probably maintain their structure and function during short-term storage, while the marked lipolysis might slow bacterial growth during this time.

Adult↗
PubMed1995

Bioactive and immunoreactive prolactin variants in human milk.

Human milk from mothers of term (T) and preterm (PT) infants was collected during early (days 2-7), mature (2-16 weeks), or late (> 16 weeks) lactation. PRL-like bioactivity (B) was measured by Nb2 cell proliferation, and PRL immunoreactivity (I) was determined by RIA. PRL activity is reported in PRL equivalents (1 PRL equivalent = 1 ng NIDDK reference material). Milk from early lactation contained significantly greater PRL-like B compared to I (T:B, 132.5 +/- 13.0; I, 83.43 +/- 12; PT:B, 195.8 +/- 56; I, 74.45 +/- 13.7). PRL-like B and I declined as lactation progressed (T mature: B, 41.74 +/- 8.9; I, 27.19 +/- 5.5; T late: B, 17.84 +/- 5.5; I, 27.33 +/- 1.8; PT mature: B, 59.85 +/- 16; I, 45.16 +/- 4.3). Milk PRL B to I ratios were consistently greater than serum B to I ratios during early lactation (milk: T, 1.4 +/- 0.3; PT, 3.6 +/- 1.3; serum: T, 1.0 +/- 0.2; PT, 0.58 +/- 0.12). During early lactation, high PRL-like B was widely distributed among several (n = 4-6) bioactive forms differing in molecular mass [8 to > 66 kilodaltons (kDa)] in T milk, but the majority of B in PT milk was detected in two or three forms. During mature and late lactation, lower PRL-like B was associated with two or three peaks (20 to > 66 kDa). A large fraction of PRL-like B (67%-84%) was associated with the phosphorylated (P-) fraction of human milk. Four immunoreactive forms (24, 30, 32, and 40 kDa) of P-PRL were identified by immunoblot analyses. Alkaline phosphatase treatment converted the 40-kDa immunoreactive P-PRL to 24-kDa PRL, increased the B of the P-fraction by 2-fold, but did not change total PRL I detected. PRL in the Concanavalin-A-retained fraction accounted for 59-69% of PRL in milk based on RIA results. No PRL-like B was detected in the Concanavalin-A-retained fraction of human milk; however, treatment of the glycosylated fraction of milk with peptide-N-glycosidase F increased thymidine incorporation by Nb2 cells 1.67-fold compared to that in controls. The results of this study show that human milk contains considerably greater PRL-like activity than previous reports based on RIA detection. The appearance and regulation of multiple bioactive PRL variants in milk throughout the course of lactation may serve as a mechanism by which milk PRL influences neonatal development.

Female↗
PubMed1994

Molecular analysis of antigens targeted by protective antibodies in rapid expulsion of Trichinella spiralis.

Rapid expulsion is a protective immune mechanism which eliminates as much as 99% of a challenge infection of Trichinella spiralis muscle larvae from the gastrointestinal tract of suckling rats. Protective monoclonal antibodies (mAbs) generated against larval excretory-secretory antigens (ESA) specifically recognize a 45-kDa glycoprotein, gp45, in addition to a distinct profile of other cross-reactive antigens that are also recognized by non-protective mAbs. Recent data indicate that protective mAbs recognize carbohydrate epitopes. To complement biochemical studies on the target(s) of rapid expulsion, we describe here the cloning and characterization of the cDNA, TspE1, which belongs to a multigene family and encodes several larval proteins in the 40-50-kDa range. A second cDNA, TspM6 encodes a 45-kDa antigen and is homologous to the published sequence of gp45. Anti-TspE1 antibodies detected antigens within beta- and gamma-stichocytes while anti-TspM6 antibodies detected antigens within alpha-stichocytes of the secretory organs of muscle larvae. Sequence analysis has provided no functional information on the encoded gene products. Neither recombinant antigen is recognized by the mAbs but native parasite molecules with peptide homology to both the TspE1 and TspM6 recombinant antigens bear the glycan recognized by the protective mAbs. These molecules are candidate targets in rapid expulsion.

Amino Acid Sequence↗
PubMed1994

Glycans as targets for monoclonal antibodies that protect rats against Trichinella spiralis.

We have investigated the role of glycans on Trichinella spiralis antigens in recognition by rat monoclonal antibodies (mAbs) which protect rat pups against challenge with the parasite. In pups born to infected dams or pups passively immunized with mAbs, antibodies eliminate a challenge dose from the intestine within hours ('rapid expulsion'). Because such dramatic protection can be afforded by mAbs, we have sought to characterize the parasite antigens they target. In this report we show that protective antibodies were unable to bind excretory/secretory (ES) antigens deglycosylated with trifluoromethanesulphonic acid (TFMS). In addition, oligosaccharides isolated from glycoproteins by alkaline hydrolysis or peptide: N glycosidase F (PNGase F) digestion were bound by protective, but not non-protective, mAbs. Glycans affinity purified with protective mAb 9D bound to all but one protective mAb. These antibodies have been shown previously to bind to the surfaces of intact larvae, indicating that the glycan is exposed on the parasite surface. Candidate glycans that may be involved in binding protective mAbs have unusual tri- and tetra-antennary structures with terminal tyvelose moieties (Reason et al., Glycobiology, 4, 000-000, 1994). Coating of the larval surface with such glycans may serve to protect the parasite and its secreted products from enzymatic attack as the parasite travels to and resides in its epithelial niche.

Animals↗
PubMed1994

Novel tyvelose-containing tri- and tetra-antennary N-glycans in the immunodominant antigens of the intracellular parasite Trichinella spiralis.

The larval stage of the intestinal nematode, Trichinella spiralis, secretes and displays on its cuticle a number of antigenically cross-reactive glycoproteins. These so-called TSL-1 antigens induce a powerful antibody response in parasitized animals. In rats, anti-TSL-1 antibodies mediate a protective immunity that expels invading larvae from the intestine. The vast majority of anti-TSL-1 antibodies are specific for glycans. Although the biological functions of TSL-1 antigens are not known, the powerful effect of glycan-specific antibodies on the intestinal survival of T. spiralis suggests that they play an important role in parasite establishment. Little is known about the structures of the glycans present on the TSL-1 glycoproteins. Recent studies have suggested, however, that the antigens contain very unusual glycans (Wisnewski, N., McNeil, M., Grieve, R.B. and Wassom, D.L., Mol. Biochem. Parasitol., 61, 25-36, 1993). Sugar and linkage analysis of the combined secreted products unexpectedly showed that a major terminal sugar is tyvelose (3,6-dideoxy-D-arabino-hexose; Tyv) which has previously been found only in certain gram-negative bacterial lipopolysaccharides. In this paper, we report the first rigorous structural study of oligosaccharides released from TSL-1 antigens by peptide N-glycosidase F digestion. Using strategies based on fast atom bombardment mass spectrometry (FAB-MS), we have discovered a novel family of tri- and tetra-antennary N-glycans whose antennae are comprised of the tyvelose-capped structure: Tyv1,3GalNAc beta 1,4(Fuc alpha 1,3)GlcNAc beta 1-. Thus a major population of TSL-1 glycans contains clusters of hydrophobic terminal structures which are likely to be highly immunogenic.

Animals↗
PubMed1992

Bile salt stimulated lipase: comparative studies in ferret milk and lactating mammary gland.

Bile salt stimulated lipase (BSSL) activity is 10-20 times higher in ferret milk than in human milk. We have used the ferret to study BSSL activity in lactating mammary gland and in mammary cells isolated by hyaluronidase-collagenase treatment followed by Ficoll gradient centrifugation. Furthermore, we have compared the characteristics of BSSL in the tissue preparations (homogenate or cells) to BSSL of ferret milk and to BSSL purified from ferret and human milk. The characteristics of BSSL in ferret mammary gland preparations and milk were similar to those of human milk BSSL--absolute requirement of primary bile salts, pH optimum of 7.5-9.0, stability at pH 3-9 and inhibition by eserine (physostigmine) and by serum. Purified ferret milk BSSL had a lower molecular weight (90kD) than did human milk BSSL (125 kD). There was an 86% homology of the N-terminal amino acid sequence between BSSL of ferret and of human milk. The marked similarity in characteristics between BSSL in ferret and human milk and the high activity of BSSL in ferret milk (520 U/mL colostrum and 250 U/mL mature milk) indicate that this species is an ideal animal model for the study of the synthesis and secretion of this digestive lipase which constitutes a significant portion (1-2%) of total milk protein.

Amino Acid Sequence↗
PubMed1991

A percutaneous liver biopsy technique in ducks (Anas platyrhynchos) experimentally infected with duck hepatitis B virus.

Aylesbury ducks (Anas platyrhynchos) chronically infected with the duck hepatitis B virus provide a useful model for studying hepadna-virus infection, replication and the effects of antiviral therapy. In these studies, it is necessary to have an effective method for obtaining repeat liver specimens for histological and molecular analyses. We have therefore developed a percutaneous liver biopsy technique which has a low rate of complications, can be performed at repeated intervals, and provides sufficient quantities of liver tissue for histological and nucleic acid hybridization analysis.

Animals↗
PubMed1990

Prevalence of hepatitis C in South Africa: detection of anti-HCV in recent and stored serum.

The prevalence of anti-HCV was studied in a cohort of 2,072 South Africans. The results were compared in selected recently collected sera and in stored sera. The serum ALT and anti-HBc were also studied as surrogate markers in this population. The following groups were tested: (a) 498 urban, black blood donors (b) 500 white blood donors (c) 500 Asian blood donors (d) 216 rural hospitalized patients (e) 358 rural mineworkers. Sera found positive by the original ELISA were retested, and reproducibly positive tests in rural black men (group d) were confirmed both by recombinant immunoblot assay and by a second ELISA. An anti-HCV prevalence of 1.2%, 0.8%, and 0.6% in urban blacks, Asians, and whites was found. Antibodies to hepatitis B core antigen were found in 42.9%, 3.4%, and 1.2% of black, Asian, and white donors, respectively; 76% of donors positive for anti-HCV were anti-HBc negative. In rural African men, 17% of stored serum samples and 9.2% of recently collected serum samples were positive for anti-HCV. In this cohort 3.84% were positive by all three assays. These results suggest that the prevalence of anti-HCV in low and high-risk South African urban blood donors is comparable to high and low prevalence areas in Europe, the United States, and Japan, but indicates a relatively high degree of exposure to hepatitis C in rural African men. The reactivity of stored, frozen sera in this population requires further investigation. In South African urban blood donors, surrogate marker testing will not expedite HCV screening.

Adult↗