Application of the IFN-gamma ELISPOT assay to quantify T cell responses against proteins.

The ELISPOT assay has been established for the direct ex vivo quantification of peptide-reactive T lymphocytes from peripheral blood mononuclear cells (PBMC). In this report we studied the ELISPOT assay conditions for the detection of T cell responses against protein antigens including Tetanus toxoid (TT), purified-protein-derivative (PPD) and a synthetic 30 mer peptide derived from influenza matrix protein (IMP) containing the HLA-A*0201-restricted 9 mer peptide epitope GILGFVFTL as a model for a viral protein. We found several aspects to be crucial for a sensitive detection of T cell responses against proteins including a pellet preincubation step and the monocyte concentration. Using optimized assay conditions specific CD4+ T cell responses against TT and PPD as well as CD8+ T cell responses against IMP can be quantified directly ex vivo from peripheral blood mononuclear cells.

Episodic angioedema with eosinophilia: precursor lesions and relevance of histology. A case report.

A 59-year-old patient suffered from persistent angioedema-like swellings and blood eosinophilia. The disease was preceded by an erythema multiforme-like exanthema and urticarial papules. Persistent eosinophilia with exclusion of its known causes and skin biopsy helped to establish the diagnosis of the benign variant of hypereosinophilic syndrome known as 'episodic angioedema with eosinophilia'. Corticosteroid therapy was successful. The therapeutic alternatives are discussed.

Dermatological signs of nasopharyngeal linguatulosis (halzoun, Marrara syndrome)--the possible role of major basic protein.

Two hours after ingestion of improperly cooked meat a German tourist in Tunisia showed coughing, hoarseness, dysphagia, anosmia, frontal headache and epistaxis. At the same time a papular non-itching exanthema developed. The nasal discharge contained nymphs of Linguatula serrata. Histological examination of the papules revealed tissue eosinophilia and 'flame figures'. Nasopharyngeal and skin signs subsided spontaneously within 10 days. The possible role of major basic protein in the pathogenesis of nasopharyngeal linguatulosis is discussed.

The structure and light-dependent transient expression of a nuclear-encoded chloroplast protein gene from pea (Pisum sativum L.).

A nuclear gene encoding a light-induced transiently expressed protein that is localized in the chloroplast has been isolated from an EMBL3 library of pea DNA. The gene is a member of a multigene family. The sequence of the gene contains the complete reading frame of previously characterized cDNA clones and two introns in the 5' region of the protein coding sequence. Primer extension and S1 nuclease studies have defined the cap site. Two TATA boxes are found 5' to the initiating methionine codon. Only a limited homology is found between the presequence of the gene and transit sequences of other previously sequenced precursors. Isolated nuclei of pea have been labelled and the in vitro synthesized transcripts analysed. The results show that the light-dependent expression of the gene family is regulated at the level of transcription.

[Modification of the internal milieu by drainage disorders following urinary diversion]. / Beeinflussung des Milieu interne durch Abflussstörungen nach Harnumleitung.

On the basis of follow-up examinations in 96 patients with diversion of urine, the effects of urinary stasis on serum electrolytes and the acid-alkali balance are demonstrated and discussed. Early correction of urinary stasis is necessary in order to avoid any impairment of the distal functioning of the tubules and the collecting tubes. The special position of serum chloride in any diversion of urine is demonstrated.

Methylation of nicotinamide in rat liver cytosol and its correlation with hepatocellular proliferation.

The changes in the activity of nicotinamide: S-adenosylmethionine methyltransferase (nicotinamide methylase) were studied in rat liver which was subjected to different rates of cellular proliferation. The cytosolic enzyme activity increased 3-4-fold in the first 24-48 h after partial hepatectomy and decreased again to the basal levels until 4 days post-operatively, whereas it remained unchanged in the livers of sham-operated animals. A single administration of thioacetamide at a dose of 50-250 mg/kg body weight, a treatment which induces hepatocellular proliferation as well, also enhanced the enzyme activity 2-3-fold 24 h after drug administration. This activity increase was associated with a marked lowering of intracellular NAD content of as much as 50% of the control levels. D-Galactosamine, a known hepatotoxic agent causing acute hepatitis in experimental animals and preventing DNA synthesis in regenerating liver, blocked the activity increase in regenerating rat liver. The rate of 1-methylnicotinamide synthesis, as measured by incubating liver slices in the culture medium supplemented with [14C]nicotinamide as a precursor, was found to be 2-4 times higher in the slices from regenerating liver and thioacetamide-treated rat liver than those from non-proliferating control liver. These results, together with our previous finding on the enhancement by 1-methylnicotinamide of the growth of cultured rat liver cells (Hoshino, J., Kühne, U. and Kröger, H. (1982) Biochem. Biophys. Res. Commun. 105, 1446-1452), support the view that nicotinamide methylase and its product, 1-methyl-nicotinamide, are involved in the control of hepatocellular DNA synthesis and proliferation.

[Experiences with lymphadenectomy in testicular tumors]. / Erfahrungen mit der Lymphadenektomie bei Hodentumoren.

The results of 48 transperitoneal, bilateral lymphadenectomies are analysed. In stage I and II we achieved a 2-year survival time of 85.2%. The operative approach is described. The findings in patients without response to the therapy are emphasized and discussed.

Regulation and characterization of L-serine: pyruvate aminotransferase in rat liver cytosol and mitochondria.

Distribution of rat liver serine: pyruvate aminotransferase between cytosol and mitochondria varies considerably with the dietary and hormonal state of animals. Feeding a high-protein diet or fasting the animals results in an increase in the enzyme activity of both fractions but more marked in the mitochondrial fraction. A low-protein diet exerts the reverse effect. A single administration of dibutyryl cyclic AMP causes a rapid elevation of the enzyme activity in both fractions, which is effectively prevented by cycloheximide, actinomycin D and cortisone. The activity in mitochondria increases with a lag of 2 h following injection of the nucleotide inducer, in contrast to the cytosol enzyme, which increases without any lag. Gel filtration and DEAE cellulose chromatography of the enzyme from both fractions revealed the similar pattern and some kinetic constants of these two types of the enzyme were not significantly different from each other. These results indicate that rat liver serine: pyruvate aminotransferase is synthesized in the extra-mitochondrial site and transfered to mitochondria.

Inhibitory effect of cortisone acetate on the stimulation of rat liver cytosol L-serrine. Pyruvate aminotransferase by dibutyryl adenosine 3,5-monophosphate.

An injection of cortisone acetate at a dose of 5 mg/100 g body weight concomitant with dibutyryl cyclic AMP prevents the increase in the activity of rat liver cytosol serine aminotransferase (L-serine:pyruvate aminotransferase, EC 2.6.1.51) elicited by the nucleotide with a lag of about 2 h. If the glucocorticoid is given 2 h prior to the nucleotide inducer, the lag disappears. The inhibitory effect of cortisone acetate gradually decays and is no longer detectable 12 h following its administration. Theophylline, insulin and glucose at doses which affect significantly the level of tyrosine aminotransferase, have not effect on the level of serine aminotransferase and on the cortisone inhibition. The inhibitory effect of the glucocorticoid on the dibutyryl cyclic AMP-mediated increase in serin aminotransferase diminishes with the age of animall. Increases in the enzyme activity by a single dose of glucagon can also be inhibited by cortisone acetate and actinomycin D as in the case with dibutyryl cyclic AMP as an inducer. The possibility of the existence of a specific inhibitory factor which is formed in response to cortisone acetate is discussed.