Biomedical subjects
Publications and source records attributed to R Lerner.
Three broad-spectrum cephalosporins (cefotetan, moxalactam, and cefoxitin) proved effective in this randomized, prospective trial for treatment of 303 surgical patients with moderately severe regional peritonitis.
Recombinant vectors carrying DNA fragments from the BamHI E region of the B95-8 Epstein-Barr virus (EBV) genome were transfected into COS-1 cells, and the transient expression of EBV-encoded nuclear antigens (EBNAs) was analyzed by using polyvalent human antisera and rabbit antibodies to synthetic peptides. Vector DNA containing two rightward open reading frames in the BamHI E fragment, BERF2a and BERF2b, induced the expression of a nuclear antigen identical serologically and with respect to size to the larger of the two polypeptides previously designated as EBNA4 in B95-8 cells. An antigen corresponding to the smaller polypeptide was induced in cells transfected with constructs that contained two neighboring reading frames, BERF3 and BERF4. This antigen also reacted with a rabbit antiserum to the synthetic peptide 203, deduced from BERF4. Thus, the findings show that the two components of the EBNA4 doublet in B95-8 cells are encoded by separate genes. The antigen encoded by BERF2a and/or BERF2b has been designated as EBNA4 and the antigen encoded by BERF3 and/or BERF4 has been designated as EBNA6. Polyvalent human antisera detected EBNA4 and EBNA6 in 9 of 11 lymphoid cell lines carrying independent EBV isolates. In the remaining two lines, either EBNA4 or EBNA6 was not detectable.
A case of visceral leishmaniasis is presented, where a pre-existing polycythemia vera obscured signs of the infection.
Because 2 patients with acute leukemia expired in septicemia with multiresistant Corynebacterium group JK (JK) the occurrence and significance of these bacteria in a hematological ward was analysed. During the following year JK was isolated in 6 other patients with acute leukemia, in 5 as a colonizing agent and in 1 as cause of an anorectal abscess. The environmental investigation with cultures from all patients, personnel and rooms in the ward disclosed heavy contamination with JK in 2 isolation rooms housing JK-colonized patients. Contamination with JK in other rooms was very sparse and there were no JK-positive cultures from personnel or other patients in the ward. Repeated environmental cultures taken after the JK-colonized patients left the isolation rooms showed sparse contamination with JK. Thus, JK strains can survive in the environment for a long time.
Detection of breast tumors during pregnancy is difficult because of hormone-influenced tissue engorgement and the tendency to limit attention to the pregnancy itself. Eight cases of breast cancer during pregnancy were seen among 12,500 pregnancies in a five-year period. All diagnoses were confirmed by surgical biopsy. Five of the eight patients detected their own tumors. Two requested abortion, but prophylactic abortion did not appear to improve outcome in the one patient with advanced disease. Seven of eight patients were between 35-40 years of age. Although based on limited experience, a plan for managing breast cancer during pregnancy is suggested. It appears that a more favorable prognosis for cure and longer periods of remission can be achieved with better breast examinations, responsible screening techniques, early diagnosis, and prompt therapy.
Autologous fibrin glue was evaluated as a sealant of the common bile duct (CBD) with the use of experimental animal models. It proved to be highly effective. Since it is a safe product that can be easily obtained from the patient's own blood, it may find useful application in most biliary tract procedures, liver transplants, and hepatectomies.
A two-site enzyme (TSE) immunoassay was developed for the quantitation of the Epstein-Barr virus (EBV)-determined nuclear antigen (EBNA) using a rabbit serum raised against a synthetic peptide derived from the BamHI K region of the viral genome. Comparison of 12 EBNA-positive and 3 negative cell lines proved that the test was EBV-specific. A dot-blot assay utilizing cloned and nick translated EBV-DNA BamHI M fragment confirmed the EBV-carrier status of the EBNA-positive lines. The results obtained with both the TSE immunoassay and dot-blot assay were in agreement with published values. In contrast to earlier reports, we could not demonstrate any correlation between the content of EBNA and the number of viral genome copies.
A 94-residue polypeptide isolated from human seminal plasma and its chemically synthesized C-terminal 28-residue segment were studied in an in vitro inhibin bioassay utilizing rat pituitary cell cultures. Both peptides have previously been claimed to have inhibin activities, and the effects on the secretion and cellular content of gonadotrophins (FSH and LH) were now assessed in the in vitro assay. No inhibition was found. After 72 h of culture, both the cellular content and the spontaneous as well as the LHRH-stimulated release of bioactive or immunoactive FSH and LH remained unaffected. Similarly, no effects were found on the storage and/or release of prolactin, growth hormone, or thyrotropin. We conclude that both the native 94-residue peptide and the synthetic replicate of its C-terminal 28-residue segment, do not influence the pituitary FSH secretion when assessed in this in vitro system.
Upper extremity lymphedema resulting from the radiation or surgical therapy of breast cancer is quite common and can approximate 25% in some series. To treat patients with this condition, a special lymphedema clinic was started in 1979. To date, 150 patients have been enrolled. They have had their lymphedema for periods of 1 month to 14 years. Treatment was either by sequential pumping using a special device (Lymphapress) or by microsurgery. The surgery consisted of multiple lymphovenous anastomoses without sutures, carried out on the medial aspect of the affected upper arm. Thus far, 102 anastomoses have been performed in 15 patients. The results of both treatment methods have been very encouraging.
Ultrasonographic evaluation of the abdominal aorta is most often done with the patient in the supine position. The right lateral decubitus position, which views the aorta through the left flank has, until now, been considered unsatisfactory for aortic evaluation. One hundred consecutive patients were prospectively examined for visualization of the aorta both through the left flank and the anterior abdomen. Twenty-one patients were then comparatively examined from the right coronal and left coronal approach. These studies showed that the aorta was clearly visualized using the left flank approach in the majority of patients (96 per cent). The combined approach yielded 99 per cent satisfactory visualization, and in a few select cases (13 per cent) the left flank was actually superior. The left flank approach was superior when directly compared with the right flank in 42 per cent of patients and comparable in 48 per cent. The right flank approach was superior in only 10 per cent.
Rats and rabbits were immunized with synthetic peptides corresponding to the VH hypervariable regions of several alpha (1----3) dextran-specific antibodies from mice to study the efficacy of synthetic peptides in the generation of site-specific anti-idiotypic reagents. Synthetic peptides were made which corresponded to the HV1, HV2, and HV3 hypervariable regions of the heavy chain of M104 (IdX+, IdI-(M104)+), HV2 of HDex 14 (IdX-), and HV3 of J558 (IdX+, IdI(J558)+). The HV1(M104) peptide sequence is found in all dextran-specific immunoglobulins examined and the HV2 and the HV3 peptides span the regions implicated in IdX and IdI expression, respectively. Sera from many rabbits and rats indicate that all five peptides are immunogenic. Antisera to HV3 peptides show excellent binding to the appropriate myeloma proteins, with antisera to the HV3(M104) peptide demonstrating little binding to proteins that differ in HV3 sequence. Antisera generated against HV1(M104) and both HV2 peptides show weak cross-reaction to the appropriate proteins; however, these sera are not idiotypic because they cross-react with immunoglobulins with very limited sequence homology. Thus, it appears that some, but not all synthetic peptides from hypervariable regions will be capable of generating antisera with useful anti-idiotypic specificities. This may reflect differences in the intrinsic antigenicity of various parts of the VH region.
Five peptides were synthesized on the basis of amino acid sequences predicted from the transformation-associated BamHI WYH region of the genome of the Epstein-Barr virus (EBV). Antisera to two peptides deduced from a 1.6-kb open reading frame in the BamHI H fragment identified an 87 000-dalton nuclear polypeptide that was present in EBV-carrying cell lines that expressed the second EBV-determined nuclear antigen (EBNA-2). This polypeptide was not detected in cell lines that carried EBV variants with a deleted BamHI WYH region or in EBV-negative cell lines. Three peptides deduced from the 1.6-kb open reading frame reacted with human EBNA-positive sera, but not with EBNA-negative sera. Following affinity purification with the peptides, two of the corresponding human antibodies also reacted with the 87 000-dalton polypeptide.
In an attempt to better understand the molecular basis of idiotypy, we have generated several site-specific antisera through immunization of animals with synthetic peptides corresponding to the (JH1) heavy chain joining segment 1 of the mouse heavy chain variable (VH) region. These anti-peptide sera identify several idiotypic determinants present on intact hybridoma and myeloma immunoglobulins. Expression of at least three of these idiotopes is correlated with the antigen specificity of the family of immunoglobulins bearing the determinant. Use of synthetic peptides may prove a powerful technique in the generation of molecularly defined antiidiotypic reagents.
Five peptides corresponding to amino acid sequences predicted from all three reading frames of the nucleotide sequence of the third internal repeat array (IR3) of the Epstein-Barr virus (EBV) genome were synthesized chemically. All five peptides elicited antipeptide antibodies in rabbits. The antiserum raised against a 14-residue copolymer of glycine and alanine gave brilliant EBV-specific nuclear staining in the anticomplement immunofluorescence (ACIF) assay, in line with the original definition of the EBV-determined nuclear antigen (EBNA) [Reedman, B. M. & Klein, G. (1973) Int. J. Cancer 11, 499-520]. Eight EBNA and EBV DNA-carrying lines showed nuclear staining with the antipeptide antibody, whereas five EBV DNA negative lines failed to stain. The staining pattern was more discretely punctate than the finely dispersed diffuse EBNA staining obtained with human antisera. Human EBV antibody-positive but not EBV-negative sera reacted with the synthetic peptide in an ELISA test. The peptide-specific antibodies were purified from the sera of healthy EBV-seropositive persons by affinity chromatography with the peptide. They gave an EBV-specific, brilliant punctate nuclear ACIF staining similar to that of the rabbit antipeptide antibodies. It was concluded that the glycine-alanine structure encoded by the IR3 region contains a native determinant of EBNA, detected by the ACIF test. Immunoblotting with the rabbit and human peptide-specific antibodies identified poly-peptides that varied between 70 and 92 kilodaltons in size in different EBV-positive cell lines, corresponding closely to a previously identified variation pattern in the size of EBNA. In addition, rabbit antipeptide antibodies identified two cellular polypeptides, 44 and 49 kilodaltons in size.
To investigate the origin of the cyclic changes in the rate of endogenous carbon-monoxide production (nCO) during the menstrual cycle, haem turnover was determined before and after chorion gonadotropic hormone-induced ovulation in six female rabbits. 14C-labelled delta-aminolevulinic acid and glycine were administered and the excretion rate of 14CO (A14CO) was measured for determination of hepatic and bone-marrow haem turnover, respectively. Carbon-monoxide production (nCO) was measured to estimate total haem turnover. After ovulation A14CO was increased significantly the first 2 h of the early labelled peak after 14C-ALA administration and was increased also at the first determination during the early peak after 14C-glycine but statistically not significantly. The total excretion of labelled CO during the period of the early labelled peak was not increased with any of these precursors for haem synthesis. On the other hand nCO was increased 34% (P less than 0.05) during the post-ovulation period. As the increase in 'unassigned' haem turnover was small and may be unaccompanied by a contemporary increase in bilirubin/CO production, it was concluded that the increase in nCO during the post-ovulation period essentially depends on increased destruction of circulating red cells in the rabbit.
Patients with newly diagnosed multiple myeloma were randomly allotted to an intermittent high-dose melphalan/prednisone (MP) treatment (120 patients) or a continuous low-dose melphalan (M) regimen (99 patients). The median observation time was 59 months (range 33-84). Response to therapy was obtained in 45% of the MP group and 31% of the M group (P less than 0.05). No significant difference in response with regard to clinical stage was noted. Median survival was 36 months in the MP group and 29 months in the M group. Survival was longer in stage I and II myeloma than in the stage III cases, at least in the MP group. The median and 5-yr survival rates in stages I and II were significantly better in the MP than in the M group. Response to therapy was associated with length of survival, median survival being 62 months in responding patients and 20 months in non-responders. The MP and M groups did not differ in this respect.
Two synthetic peptides corresponding to residues 1-20 and 10-20, respectively, of one type of a cellular protein called "p53" have been linked to a carrier protein and injected into rabbits to raise antibodies. The antibodies obtained were capable of reacting with the native protein, as judged by an enzyme-linked immunosorbent assay, protein A-linked staining of immunoblots after NaDodSO4 gel electrophoresis, and immunoprecipitation. The immunoassay titers against the protein were lower for these antibodies than for antisera derived from immunization with purified p53. However, staining with the immunoblot method showed that the antipeptide antibodies against p53 were uniquely specific. The data suggest that at least two different types of p53 molecules occur. The cellular protein previously isolated from human cells transformed by Epstein-Barr virus and from murine tumors induced by methylcholanthrene appears to be larger than the p53 reported in relation to simian virus 40- or adenovirus-transformed cells and to some other tumors. Some interrelationships have not been excluded, but it is clear that the two protein molecules do not behave identically. The reactions of the antipeptide antibodies with the intact protein have implications in regard to protein conformations. The strict specificities of such antibodies allow the generation of distinct sets of reagents useful for quantitation, purification, and cloning.
In 17 patients requiring regular hemodialysis, inter- and intradialytic hemoglobin catabolism measured as endogenous carbon monoxide production (VCO) were studied before (9 patients) and after (8 patients) the introduction of water treatment. Before the water treatment, VCo increased significantly from inter- to intradialytic periods whereas insignificant changes were noted after the water treatment. Thus water treatment leads to less destruction of erythrocytes in connection with hemodialysis. However, hemolysis associated with hemodialysis does not play a major role as a cause of anemia in these patients.