Dihydropyrimidine dehydrogenase deficiency caused by a novel genomic deletion c.505_513del of DPYD.

Dihydropyrimidine dehydrogenase (DPD) deficiency is an autosomal recessive disorder of the pyrimidine degradation pathway. In a patient presenting with convulsions, psychomotor retardation and Reye like syndrome, strongly elevated levels of uracil and thymine were detected in urine. No DPD activity could be detected in peripheral blood mononuclear cells.

Analysis of severely affected patients with dihydropyrimidine dehydrogenase deficiency reveals large intragenic rearrangements of DPYD and a de novo interstitial deletion del(1)(p13.3p21.3).

Dihydropyrimidine dehydrogenase (DPD) deficiency is an infrequently described autosomal recessive disorder of the pyrimidine degradation pathway and can lead to mental and motor retardation and convulsions. DPD deficiency is also known to cause a potentially lethal toxicity following administration of the antineoplastic agent 5-fluorouracil. In an ongoing study of 72 DPD deficient patients, we analysed the molecular background of 5 patients in more detail in whom initial sequence analysis did not reveal pathogenic mutations.

In situ chemical cleavage of proteins immobilized to glass-fiber and polyvinylidenedifluoride membranes: cleavage at tryptophan residues with 2-(2'-nitrophenylsulfenyl)-3-methyl-3'-bromoindolenine to obtain internal amino acid sequence.

Proteins immobilized to glass-fiber supports and polyvinylidenedifluoride membranes are cleaved in situ with a tryptophan residue-specific reagent, 2-(2'-nitrophenylsulfenyl)-3-methyl-3'-bromoindolenine. Protein fragments can be eluted from polyvinylidenedifluoride membranes after in situ digestion and electrophoresed and electroblotted to new polyvinylidenedifluoride membranes for subsequent sequence determination of selected bands. Five proteins (bovine serum albumin, ovalbumin, beta-casein, beta-lactoglobulin, and myoglobin) of known sequence containing one to three tryptophan residues each were used as model substrates to evaluate the specificity and extent of cleavage.

Immunologic and clinical status of blood donors with subnormal levels of IgG2.

To determine the immunologic and clinical status of individuals from a general population with subnormal levels of IgG2, we prospectively studied 37 of 312 blood donors with low IgG2 levels identified among 8015 donors. We examined (1) G2m(23) allotypes, (2) levels of other IgG subclasses and immunoglobulin classes, (3) composition of peripheral leukocyte populations, (4) responses to two carbohydrate antigen vaccines, (5) in vitro secretion of IgG subclasses by isolated lymphocytes after mitogen stimulation, and (6) clinical histories. We found that most (90%) individuals with subnormal IgG2 levels had G2m(23)- allotypes, whereas only 30% of the unselected donors had G2m(23)-.

Production, characterization and use of five monoclonal antibodies to human IL-4.

We describe five murine monoclonal antibodies that are specific for human IL-4. At least three spatially distinct epitopes on the hIL-4 molecule are recognized by this panel of antibodies which allowed development of a sensitive sandwich EIA specific for hIL-4. The EIA is capable of detecting 200 pg/ml of hIL-4 and exhibits no detectable crossreactivity to seven other human cytokines examined.