Identification of twenty-two candidate markers for human osteogenic sarcoma.

Since osteogenic sarcoma (OGS) predominantly affects children, its etiology and progression may be determined more by genetic than environmental factors. A few genes have been associated with OGS, however, their value in the diagnosis and/or prognosis of the disease remains poor. Evidently, more markers need to be identified for improving management of patients with OGS. To identify potential genetic markers for OGS, we have extended preferential amplification of coding sequences (PACS) to screen multiple samples simultaneously. The extended method is termed multi-PACS. Multi-PACS was applied between a normal osteoblast and four OGS-derived cell lines to identify differentially expressed coding sequence tags (dCST) that identified 145 dCSTs. Subsequently, differential mRNA expression was validated for a chosen subset of 22 dCSTs. These chosen dCSTs include among others cyclins D and E, two cyclin dependent kinases, two other kinases, transcription factors E2F4, E2F5, and p130, a DNA repair gene, a gene for the signalosome subunit, and potential guanine nucleotide binding factors. We infer that these genes could be so easily identified because PACS preferentially identifies coding instead of non-coding sequences. We also infer that these genes identify signaling pathways pertinent to OGS. mRNA expression profile of these 22 genes/dCSTs generated distinct expression signature of the OGS-derived cell lines suggesting that further work on clinical samples with these dCSTs will yield valuable information for OGS. We conclude that these 22 genes/dCSTs are candidate markers for OGS.

Autoimmune hemolytic anemia caused by IgG lambda-monotypic cold agglutinins of anti-Pr specificity after rubella infection.

BACKGROUND: In postinfection cold agglutinin (CA) disease, a relation between CA specificity and the underlying infectious agent has been observed. The induction of anti-I by Mycoplasma pneumoniae and that of anti-i by EBV are well-established examples. CASE REPORT: A 5-year-old boy developed severe hemolytic anemia after serologically ascertained rubella infection. Hemolysis was caused by high-titer CAs, which were analyzed by absorption and elution with sialidase-treated RBCs and hemagglutination-inhibition experiments. RESULTS: After elimination of normal anti-I and anti-T, the predominant CA was found to be an IgG lambda autoantibody with anti-Pr(1) specificity. CONCLUSION: This case seems to be of interest because it is the first report of severe CA-induced hemolysis after rubella infection, it is the first description of an IgG lambda-monotypic CA, and, along with previous case reports (three established and three suspected cases), it indicates a relationship between rubella infection and the CA specificity anti-PR:

First workshop for detection of heparin-induced antibodies: validation of the heparin-induced platelet-activation test (HIPA) in comparison with a PF4/heparin ELISA.

BACKGROUND: No data exist regarding the inter-laboratory reproducibility of the heparin-induced-platelet-activation (HIPA) test, the most widely used functional assay in Germany for the detection of heparin-induced thrombocytopenia (HIT) antibodies. METHODS: Nine laboratories used an identical protocol to test eight different sera with the HIPA test. Five laboratories also tested the sera with a platelet factor 4 (PF4)/heparin-complex ELISA. Cross-reactivity with danaparoid-sodium was assessed using 0.2 aFXa units instead of heparin in the HIPA test. RESULTS: Two of nine laboratories had no discrepant HIPA test results. Four laboratories differed in one sample, one reported two discrepant results, and two laboratories reported more than two discrepant results. Cross-reactivity with danaparoid-sodium test results differed among laboratories. PF4/heparin ELISA results were identical in all five laboratories. CONCLUSION: The HIPA test requires strict quality control measures. Using both a sensitive functional assay (HIPA test) and a PF4/heparin ELISA will allow detection of antibodies directed to antigens other than PF4/heparin complexes as well as detection of IgM and IgA antibodies with PF4/heparin specificity.

[Severe neonatal alloimmune thrombocytopenia with delayed antibody detection]. / Schwere neonatale Alloimmunthrombozytopenie mit verzögertem Antikörpernachweis.

Neonatal alloimmune thrombocytopenia (NAIT) is caused by maternal immunisation against a paternal antigen on fetal platelets. The antigen involved in the majority of cases is HPA-1 a (PIA1). Usually circulating platelet alloantibodies are detectable in the mother. In this report, we present a thrombocytopenic newborn with severe hemorrhagic diathesis due to materno-fetal HPA-1 a (PIA1) incompatibility. Platelet antibodies could initially not be demonstrated in the mother's serum but became detectable after four weeks. Because of the severe and protracted course of the disease, repeated platelet substitution was necessary throughout the first two months of life.

Incidence and persistence of anti-Kell after transfusion of Kell-positive blood.

116 Kell-negative patients who had received at least one Kell-positive blood unit were tested about 3 and 12 months after transfusion. Antibody screening was performed by tube test and by gel centrifugation; both methods yielded identical results. After 3 months, anti-Kell was detected in 11 (9.5%) patients. Antibody titers using K+, k+ cells ranged from 1:1 to 1:128 with a predominance of low titers. Titers using K+, k- cells tended to be higher by one step; one anti-Kell could only be demonstrated with K+, k- cells. After about 12 months, anti-Kell could no longer be detected in 5 of the 11 patients (45.5%).

Efficient antibody screening using gel centrifugation.

In a prospective study including 6,674 sera, antibody screening was performed (1) by tube test (saline/albumin/antiglobulin), (2) by gel centrifugation (Diamed) using two tests, saline/room temperature and antiglobulin/37 degrees C, and (3) by gel centrifugation using a single test antiglobulin (IAT)/ room temperature (RT). By gel centrifugation (saline RT + IAT 37 degrees C), compared to tube test, 40% more Coombs-reactive antibodies but 60% fewer cold-reactive antibodies were detected. Similar results were obtained by gel centrifugation using a single test IAT RT instead of the two tests saline RT + IAT 37 degrees C. A case of delayed hemolytic transfusion reaction was observed caused by anti-C demonstrable only by gel centrifugation. [table: see text]

Activity of glycopeptides against vancomycin-resistant gram-positive bacteria.

Gram-positive bacteria resistant to vancomycin are rare; but they include members of the genera Leuconostoc, Lactobacillus, and Pediococcus, as well as recently emerging vancomycin-resistant strains of Enterococcus faecium and Enterococcus faecalis. Vancomycin, teicoplanin, and several vancomycin derivatives were tested for their activities against vancomycin-resistant gram-positive bacteria. Vancomycin-resistant E. faecium and E. faecalis were generally cross-resistant to other glycopeptides, but some N-substituted vancomycin derivatives were active against the resistant strains, with MICs of 2 to 32 micrograms/ml. These vancomycin derivatives also had significant levels of activity against intrinsically vancomycin-resistant organisms such as Leuconostoc sp. While vancomycin resistance in E. faecium and E. faecalis was inducible, resistance in members of the genera Leuconostoc, Lactobacillus, and Pediococcus appeared to be expressed constitutively. Antibody to a vancomycin-induced membrane protein found in membranes of resistant enterococci did not detect a cross-reacting protein in other vancomycin-resistant species.

Synthesis and antibacterial evaluation of N-alkyl vancomycins.

Over eighty N-alkyl vancomycins were synthesized by reductive alkylation of vancomycin with the appropriate aldehydes. The N-alkyl vancomycins exhibit greater antibacterial activity than the corresponding N-acyl vancomycins and the parent antibiotic. Some of these semisynthetic vancomycins are five times more active than vancomycin. The N-alkyl vancomycins also show longer elimination half-lives in rats than vancomycin.

Synthesis and antibacterial activity of N-acyl vancomycins.

Several glycopeptides containing N-acyl groups have been isolated recently. We undertook the synthesis of N-acyl vancomycins, using the active ester method. The in vitro and in vivo antibacterial activity were evaluated, and structure-activity relationship of this series of semisynthetic vancomycins is discussed.