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Biomedical subjects

D Barnett

Publications and source records attributed to D Barnett.

At least 91 records · Page 5Linked to original sources

The interleukin-2 receptor and its expression in the acute leukaemias and lymphoproliferative disorders.

The expression of the interleukin-2 receptor (IL-2R) was investigated in a total of 117 cases of acute leukaemia and lymphoproliferative disorder. The results confirm reports that the interleukin-2 receptor can be expressed on cells of myeloid lineage and in cases of Hairy Cell Leukaemia. They extend knowledge of the expression of the IL-2R in cases of lymphoproliferative disorders.

Acute Disease↗

Studies on the subunit structure of textilotoxin, a potent neurotoxin from the venom of the Australian common brown snake (Pseudonaja textilis).

Textilotoxin is a presynaptic neurotoxin from the venom of the Australian common brown snake, Pseudonaja textilis. It has the highest lethality and is structurally the most complex of any known snake venom neurotoxin. It was resolved into its five non-covalently linked subunits in a single step by reverse-phase HPLC. Two of the subunits were identical. The N-terminal amino-acid sequence and amino-acid composition of each subunit were determined. Subunit A was the only one found to possess phospholipase A activity. Separation of textilotoxin into its subunits was reversible and reformed textilotoxin had the same Mr and lethality in mice as the native toxin. Experiments with various unnatural combinations of subunits have led to interesting variations in lethality and Mr of the resulting complexes.

Amino Acid Sequence↗

Pseudonajatoxin b: unusual amino acid sequence of a lethal neurotoxin from the venom of the Australian common brown snake, Pseudonaja textilis.

The complete amino acid sequence of pseudonajatoxin b, a basic neurotoxin from the venom of the Australian common brown snake, Pseudonaja textilis, was determined by automated Edman analysis of the reduced carboxymethylated polypeptide and of peptides derived by digestion of it with Staphylococcus aureus V8 proteinase. Pseudonajatoxin b consists of a single polypeptide chain of 71 amino acids with Mr 7762. The amino acid sequence showed considerable homology with postsynaptic long neurotoxins, but there were striking differences. Pseudonajatoxin b displayed relatively high lethality, LD50 15 micrograms/kg in mice.

Amino Acid Sequence↗

Lectins and the radioallergosorbent test.

An investigation into the possible role of lectin binding of IgE in RAST of legume and wheat extracts is reported. Lectins from pea, broad bean, lentil, jack bean, soybean, peanut, and wheat germ were coupled to RAST discs. The discs were pretreated with lectin-specific sugars in an attempt to inhibit RAST with sera from 11 sensitive patients. In all cases, RAST was almost unaffected by the inhibitory sugars, indicating that nonimmune binding of IgE by lectins in legume or wheat RAST was not significant when RAST was carried out with allergens bound to the usual paper discs. IgE contains binding sites for all the lectins examined, and five of the sera had high total IgE greater than 300 IU/ml. It is suggested that competition by IgG and other serum glycoproteins may explain the lack of effect by the added sugars. All sera contained endogenous glucose at a level of about 3 mmol/L that may have accounted for some self-inhibition of the lectin binding by pea, broad bean, lentil, and jack bean lectins. There was, however, significant immune binding of some of the lectins by specific IgE, and it is concluded that these lectins may be important in expression of IgE-mediated allergic responses.

Arachis↗

Allergenic cross-reactions among legume foods--an in vitro study.

The specific IgE binding by protein extracts of 11 food legumes, including soybean, was examined by RAST and RAST inhibition. Sera from 15 peanut-sensitive patients were, with very few exceptions, positive in the RAST to all the legumes. RAST-inhibition testing of each extract against RAST discs of the other legumes indicated considerable cross-reactivity of IgE binding between the legumes. Cross-allergenicity was demonstrated to be most marked between the extracts of peanut, garden pea, chick pea, and soybean. The results have important implications for selection of effective hypoallergenic diets and for the diagnosis of patients hypersensitive to foods.

Adolescent↗

Transferrin receptor expression in the leukaemias and lymphoproliferative disorders.

The expression of the transferrin receptor (TfR) was studied in the acute leukaemias and lymphoproliferative disorders by means of indirect immunofluorescence and/or the enhanced alkaline phosphatase anti-alkaline phosphatase (APAAP) techniques using monoclonal antibodies to the receptor. A total of 174 cases of acute leukaemia and lymphoproliferative disorder were studied. The results indicate that the receptor is expressed with increased density in the majority of positive cases of acute leukaemia. The lymphoproliferative disorders, however, only expressed the receptor in a minority of cases and did so with weak density. It is proposed that this weak expression in the lymphoproliferative disorders may be of use as an indicator of an increase in cell activity.

Acute Disease↗

Granular common acute lymphoblastic leukaemia in adults: a morphological study.

2 cases of acute lymphoblastic leukaemia characterised by the presence of cytoplasmic inclusions morphologically similar to azurophil granules are described. Azurophil granulation of blasts is one of the cardinal features which differentiate acute myeloid from acute lymphoblastic leukaemia. Although such granulation of lymphoblasts has caused diagnostic confusion in the past, we found that the granules could be distinguished from myeloid azurophil granules both morphologically and by their characteristic cytochemical staining reactions. They were negative for peroxidase/sudan black and chloroacetate esterase, but gave coarse scattered granular positivity for both acid phosphatase and alpha-naphthyl acetate esterase. Both the electron microscopic appearance of the granules and their cytochemical staining reactions suggested that they were lysosomes. Granular ALL does not appear to be associated with any morphological subtype or karyotype; but is strongly associated with the common ALL phenotype. Its prognostic significance remains uncertain. It occurs more frequently than the small number of previous reports might suggest and, given the potential for misdiagnosis, should be more widely recognised.

Adult↗

Peripheral blood lymphocyte subpopulations in patients with primary proliferative and secondary polycythaemia.

Peripheral blood lymphocyte subpopulations were measured in 18 patients with primary proliferative polycythaemia and 13 patients with secondary polycythaemia. A decrease in numbers of suppressor T lymphocytes and an increase in the helper:suppressor T lymphocyte ratio was found in those with primary polycythaemia compared with normal subjects and patients with secondary polycythaemia. If other causes of an increased helper:suppressor ratio are excluded this variable may be useful in confirming the myeloproliferative nature of patients with erythrocytosis.

Adult↗

Allergenic cross-reactivity of egg-white and egg-yolk proteins. An in vitro study.

The radioallergosorbent test (RAST) and RAST inhibition test were used to examine cross-allergenicity amongst the major hen's egg-white and egg-yolk proteins. Using ovalbumin as a reference allergen to compare cross-reactivity, it was apparent that the proteins conalbumin, ovomucoid and lysozyme substantially inhibited binding to ovalbumin discs of IgE in the sera of patients clinically hypersensitive to egg. The converse situation with conalbumin, ovomucoid and lysozyme on the discs and ovalbumin as the inhibitor also resulted in significantly decreased levels of IgE binding to the proteins on the discs. It was also demonstrated that cross-reactions occurred between ovalbumin and the yolk protein, apovitellenin I. Cross-reaction was also observed surprisingly when egg lysozyme was on the disc and the milk protein allergen alpha-lactalbumin was used as the inhibitor. The demonstration of cross-reaction between all of these proteins may signify that there are a number of common allergenic determinants on these egg proteins, thus providing a molecular basis for the phenomenon of cross-reactivity.

Adolescent↗

Partial characterization of an allergenic glycoprotein from peanut (Arachis hypogaea L.).

A concanavalin A-reactive glycoprotein allergen has been isolated from peanut (Arachis hypogaea). The allergen was separated by affinity chromatography and purified by gel permeation and ion-exchange chromatography. The monomeric molecular weight is 65,000 and the pI is 4.6. The presence of one cysteine residue per molecule results in some dimer formation. Concanavalin A-reactive glycoprotein is a potent allergen for peanut-sensitive patients in both in vivo and in vitro tests. It is allergenically stable, on in vitro examination, at temperatures of up to 100 degrees C and over the pH range 2.8-10. Removal of the carbohydrate moiety failed to eliminate the allergenicity. Concanavalin A-reactive glycoprotein is identified in the crossed immunoelectrophoretic pattern as a major antigen of peanut protein extract but its structural characteristics indicate that it is probably not a component of the major storage-protein complex, arachin.

Allergens↗

Skin test, RAST and clinical reactions to peanut allergens in children.

One-hundred-and-four children were skin-tested with four peanut-allergen preparations, a commercial extract, extracts of raw and roast peanuts prepared by NH4HCO3 extraction, and a wheatgerm lectin-reactive glycoprotein obtained by affinity chromatography. The presence of symptoms after ingestion of peanut or peanut products was also recorded. The roast allergen extract provided the greatest specificity with eight symptomatic children having a positive skin test and only one positive skin-test reaction in an asymptomatic child in the group of 104 children tested. Despite differences in the incidence of skin-test reactions there was a strong correlation between raw, roast and commercial RAST suggesting common allergens were being identified by circulating IgE. Clinical sensitivity was observed particularly in younger children with 75% of the children being under 4 years of age. A positive roast skin test or a RAST test adds confirmation to the clinical history of allergic reactions to peanuts.

Adolescent↗

Lymphocyte subpopulations in patients with hydroxocobalamin responsive megaloblastic anaemia.

Lymphocyte subpopulations and intrinsic factor and gastric parietal cell antibodies have been measured in 23 patients with megaloblastic anaemia who responded to treatment with hydroxocobalamin. The ratio of helper (OKT4) to suppressor (OKT8) lymphocytes was significantly increased in patients with intrinsic factor antibody compared with those who lacked the antibody. No such correlation was found for gastric parietal cell antibody. Alterations in the lymphocyte helper to suppressor (OKT4:OKT8) ratio may be associated with pernicious anaemia.

Anemia, Macrocytic↗

Allergens in the white and yolk of hen's egg. A study of IgE binding by egg proteins.

The radioallergosorbent test (RAST) was used to compare the IgE binding of egg white and yolk, and allergenic proteins were detected by immunoelectrotransfer ('Western blotting'). The main allergens were found in egg white, but for a large proportion of the egg-sensitive patients, yolk contained specific IgE-binding constituents. For blood sera from 36 patients, there was a positive correlation between the results of RAST for egg white and for yolk. Lysozyme was found to be an allergen for some patients. The effect of heating on the allergenicity of egg white was examined and the allergenicity of hen egg white was compared with that of a duck egg. The allergens in yolk were associated with each of the three yolk fractions, and several of the proteins in the low-density lipoprotein fraction bound IgE.

Adolescent↗

The toxicological evaluation of tallysomycin s10b--biosynthetic tallysomycin derivative.

Tallysomycin S10b, a biosynthetic derivative of tallysomycin B, was subjected to intravenous toxicologic studies in mice and dogs. LD50 and LD10 values from lethality studies in mice were utilized to establish dose levels for single and five daily dose toxicity studies in mice and dogs. Nephrotoxicity was the most consistent and prominent drug-related alteration in single and multiple dose studies in both species and was considered the dose limiting toxicity. Other toxicities included pulmonary toxicity, lymphopenia and necrosis of extremities in mice and dogs and testicular degeneration and focal vacuolation of adrenal cortical cells in the dog. Bleomycin was administered to dogs as a reference control agent at a single dose of 270 mg/m2 which was approximately equivalent to the highest tallysomycin S10b dose of 240 mg/m2. Tallysomycin S10b showed a greater nephrotoxic potential than bleomycin at earlier time periods. However, at termination there was no meaningful difference in the degree of chronic nephrotoxicity. The pulmonary toxicity of both drugs was comparable.

Animals↗

Stimulation of ornithine decarboxylase activity by luteinizing hormone in rat testicular interstitial cells in vitro is age dependent.

The developmental pattern of ornithine decarboxylase (ODC) responsiveness to luteinizing hormone (LH) in isolated rat testicular interstitial cells in vitro was examined and correlated with testosterone production by the same cells. LH caused a 60-100% stimulation of ODC activity in cells from 60-day-old rats but produced no response in cells from 30, 37, 41, 50 and 55-day-old animals. Interstitial cells from 25-day-old rats responded with a moderate (40%) but statistically significant enhancement of ODC activity to the highest LH dose (100.0 ng/ml) only. Testosterone production by control cells was low until day 41 (0.15-0.30 ng/10(6) cells per 4 h), and then markedly increased to adult levels (2.12 +/- 0.03 10(6) cells per 4 h). LH in all concentrations (0.1 - 100.0 ng/ml) employed caused a consistent 4 to 7-fold stimulation of testosterone production in interstitial cells at all ages studied. This study shows age-dependent stimulation by LH of ODC activity in rat testicular interstitial cells in vitro and no apparent correlation with testosterone production by the same cells.

Aging↗

Multiplicity of allergens in peanuts.

Crude peanut protein fractions from raw and roasted peanuts were examined in the RAST with 10 sera from patients showing clinical peanut sensitivity. The radioactive uptake results, which were generally high, did not reveal any distinguishable pattern. Two commercially available peanut proteins, peanut lectin and phospholipase D, gave poor RAST responses. Three purified peanut proteins, alpha-arachin, conarachin I, and concanavalin A-reactive glycoprotein, all gave significant RAST results that were generally lower than those obtained with the crude extracts. The extent of RAST inhibition obtained with these materials was inversely related to their abundance in the total peanut protein. Crossed immunoelectrophoresis with extracts from raw and roasted peanut indicated the presence of 22 and 10 anodically migrating antigens, respectively. Sixteen IgE binding antigens were revealed for raw peanut and seven for roasted peanut after incubation with a mixed serum from the 10 patients in crossed radioimmunoelectrophoresis (CRIE) using 125I-labeled anti-IgE. CRIE plates treated with individual serum samples showed that all the patients had specific IgE for the major antigen peak, which has been tentatively identified as alpha-arachin. This major storage protein of peanut, which is known to be particularly heat resistant; may be of greater clinical significance than its apparently low RAST activity would seem to indicate.

Adolescent↗