Stroke in Icelandic patients with hereditary amyloid angiopathy is related to a mutation in the cystatin C gene, an inhibitor of cysteine proteases.

Cystatin C is an inhibitor of lysosomal cysteine proteases and consists of 120 amino acids. A variant of cystatin C lacking the first NH2-terminal residues and having one amino acid substitution at position 68 forms amyloid deposits mainly in the walls of brain arteries, causing fatal strokes in Icelandic patients with familial cerebral hemorrhage secondary to a form of an autosomal dominant amyloidosis. To understand the molecular basis of the genetic defect, the gene encoding cystatin C was isolated from genomic DNA libraries made from normal tissue and the brain of an Icelandic patient with hereditary cerebral hemorrhage with amyloidosis (HCHWA-I). The data indicate that the cystatin C gene encodes a polypeptide of 146 amino acids, of which the first 26 correspond to a secretory peptide signal sequence. The gene contains two intervening sequences that interrupt the coding region at amino acids 55 and 93. Comparison with genes encoding salivary cystatins and kininogen proteins show sequence homology and conservation of exon-intron structure. Except for a mutation in the second exon (CAG instead of CTG in the normal gene, resulting in the substitution of glutamine for a leucine residue), the gene cloned from the brain of the Icelandic patient is identical to the normal cystatin C gene. Thus, HCHWA-I is the first familial type of amyloidosis related to a point mutation in a gene encoding for an inhibitor. The mutation in the structural gene encoding cystatin C appears to be the primary defect in this inherited disorder causing amyloid fibril formation and accumulation followed by cerebral hemorrhage.

Human monoclonal macroglobulins with specificity for Klebsiella K polysaccharides that contain 3,4-pyruvylated-D-galactose and 4,6-pyruvylated-D-galactose.

Two human IgM myeloma proteins, IgMWEA and IgMMAY, were found to react with agar and Klebsiella polysaccharides that contain pyruvylated D-galactose (DGal). Quantitative precipitin data and precipitin inhibition studies with methyl alpha- and beta-glycosides of 4,6-pyruvylated-D-galactose showed their combining sites to be different, although each was directed against the pyruvylated-D-Gal, one reacting most specifically with Klebsiella polysaccharides with terminal nonreducing beta-linked 2,4 pyruvylated-D-Gal, whereas the other reacted equally well with Klebsiella polysaccharides that contain 3,4 beta-linked and 4,6 alpha-linked terminal nonreducing pyruvylated-DGal. Inhibition studies showed that both sites are directed toward one of the two space isomers of 3,4- or 4,6-pyruvylated DGal, the form in which the methyl group of the pyruvate is equatorial, or endo, and its carboxyl group axial, or exo, to the plane of the acetal ring. Coprecipitation studies showed the combining site of IgMWEA to be located on an (Fab')2 fragment and not on the (Fc)5mu fragment. The monoclonal peak in the serum of IgMMAY was specifically precipitated by Klebsiella polysaccharide. Myeloma proteins with specificities of this type may occur with reasonable frequency in humans and may be a consequence of clonal expansion from inapparent infection, carrier states, or disease produced by various Klebsiella organisms.

Acute lymphoblastic leukemia. Its occurrence with hypereosinophilic syndrome' and bilateral spontaneous pneumothorax.

A patient with acute lymphoblastic leukemia had an unusual prodrome, including clinical features of "hypereosinophilic syndrome," pulmonary infiltrates, and bilateral spontaneous pneumothorax, which preceded the onset of leukemia by four months. The mechanism for the production of eosinophilia may have been related to the production of eosinopoietic factors by the leukemic cells.

Polymorphism of human immunoglobulin VH genes: a possible marker of autoimmune disease.

Human DNA from 11 individuals was analysed by Southern blot for the immunoglobulin genes coding for the heavy chain variable region (VH). The analysis included two probes detecting the genes of subgroup VHII and VHIII. The VH genes pattern shows very little polymorphism whereas the VHIII genes showed a significant polymorphism. When DNA from four patients with Graves' disease was analysed, a VH band was found in DNA of all patients analysed, and of 50 per cent of SLE patients, whereas only 36 per cent of healthy people contained this VH band. This may serve as a new tool to study genetic markers of autoimmune diseases.

Systemic lupus erythematosus in 49 Israeli males: a retrospective study.

Forty-nine male SLE patients, diagnosed and followed in seven medical centers in Israel between 1954 and 1983, were studied and analyzed retrospectively in order to determine whether the disease in males was clinically different from that reported in females both in Israel and in the world literature. The primary clinical and laboratory manifestations, the severity of the disease at the onset or at any time during the course of the disease, and the 1-15 year survival rates were not different from those described before in female SLE, although neurological involvement, nephritis, thrombocytopenia, vasculitis and hepatosplenomegaly were more prevalent in our series. However, more than half of the male patients (53%) had a benign course of disease characterized by long remissions requiring minimal or no medication. Long-term remission of serious renal involvement was observed completely in 14 and partially in 5 out of 33 patients. These results suggest that the male sex might alter the clinical course of SLE.

Case report: fatal seronegative rickettsial infection diagnosed by the polymerase chain reaction.

A previously healthy man presented with a five day history of high fever and headache, later followed by rash and the appearance of jaundice. On the second hospital day, he suddenly developed seizures, lapsed into a coma, and died. Polymerase chain reaction (PCR) amplification revealed a 434 base pairs DNA fragment common to the genome of typhus and spotted fever group rickettsiae in the patient's blood (estimated at about 1 x 10(2) organisms/ml), and to a lesser degree in the cerebrospinal fluid. However, serological tests for rickettsiae remained negative. PCR techniques may confirm the diagnosis at an early stage, even though the rickettsemia may be minimal and the patient seronegative.

Malabsorption following aorto-femoral bypass.

A case of malabsorption which developed after aortofemoral bypass surgery is described. Barium enema showed ischemic changes, and postoperative arteriography revealed marked reduction of the peripheral vascular bed of the mesenteric arteries. Malabsorption probably developed as a consequence of mesenteric ischemia, aggravated by the surgical procedure, the so-called aorto-iliac steal syndrome.

Attenuation of anaphylactic shock and related mortality in guinea-pigs after administration of a potent protein kinase inhibitor, K252a.

We examined whether protein kinases have a role in the expression of anaphylactic shock (AS). Guinea-pigs sensitized to ovalbumin were administered i.p. saline (control) or 10 micrograms/kg K252a, a potent protein kinase inhibitor, 30 min before challenge. The development of AS and mortality was observed for the next 2 h. In the K252a-treated group the incidence of AS fell to 53% from 100%, the maximum intensity was 62% less than the control, and mortality dropped to 16% from 50% of the animals. We suggest that protein kinases are involved in the expression of AS, and that inhibitors of these enzymes may protect against the symptoms of AS and allergy.

Nitric oxide: a mediator in anaphylactic shock in guinea-pigs.

In this study we show that the pathophysiology of anaphylaxis includes generation of nitric oxide (NO), a very powerful, short-acting vasodilator. Guinea-pigs sensitized to ovalbumin were treated with 200 microgram/kg diphenylene iodonium (DPI), and NO synthase inhibitor, prior to antigen challenge. Mortality following the challenge fell from 71 to 39% (p < 0.001, n = 59). In the Langendorff preparation perfused isolated hearts from sensitized guinea-pigs were challenged to initiate cardiac anaphylaxis. The coronary flow rate (CFR), a direct reflection of coronary arterial resistance, was reduced by antigen challenge to 56 +/- 4% (n = 16) of the basal rate. DPI (2 micrograms/ml) intensified the antigen-induced fall in CFR to 13 +/- 3% of control (p < 0.005, n = 5), and the false substrate for NO, L-N-methylarginine, to 37 +/- 3% (p < 0.05, n = 4). Sodium nitroprusside (SNP), a NO generator, raised the basal CFR by 46% (from 11.2 +/- 1.7 ml/min to 16.3 +/- 1.9 ml/min) and blunted the antigen-induced fall in CFR. Paradoxically, DPI, which can inhibit flavoprotein enzymes other than NO synthase, potentiated the vasodilator effect of SNP, raising the basal CFR by 116%. Together these results strongly indicate that the vasodilator NO is generated in anaphylaxis. However, whereas in the heart it may function as a counterweight to the vasospasm of the coronary arteries, in the intact animal it appears to be a major contributor to the potentially lethal hypotension of anaphylactic shock.

[Subclinical renal involvement in rheumatoid arthritis].

No evidence of renal involvement was found in 104 patients with rheumatoid arthritis in routine laboratory tests, including serum creatinine, urea, uric acid, sodium, potassium, calcium, phosphorus, and urinalysis. In view of recent publications (1-9) which report renal involvement in rheumatoid arthritis, we studied 16 patients of our group (nonrandomized, 3 men and 16 women, average age 55.4 years, average duration of disease 11.9 years). We examined creatinine clearance, urinary excretion of alpha-2 microalbumin, beta-2 microglobulin, cystine, and urine concentration and acidity after a 10-hour fast. 10 patients had disturbances in 1 or more of the functions examined, in 9 of whom tubular functions were involved. In 6 there was no evidence of renal involvement. There was no correlation between renal involvement and past or present therapy, but there were direct correlations between renal involvement, duration of disease and age. Thus we found evidence for subclinical renal damage not revealed by routine laboratory tests in patients with rheumatoid arthritis. This damage should be taken into consideration when operation, examination with contrast material, or treatment with other nephrotoxic agents are being considered in these patients.