[Case reports of tularemia].
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Administration, Oral↗
Biomedical subjects
F W Hirsch
Publications and source records attributed to F W Hirsch.
[Differential diagnosis of tularemia].
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Animals↗
Demonstration of estrogen and progesterone receptors as well as Ki-67 and p-145 antigens in single tumor cells from blood and pleural effusions using a slide assay.
We describe a slide assay that allows the demonstration of antigens localized in the nucleus from isolated white blood cells as well as from single tumor cells derived from malignant effusions. With the antibodies Ki-67 and anti-p-145 an increased rate of nuclear and nucleolar staining resulted in cells from highly malignant lymphomas. An almost identical reaction was obtained when tumor cells from malignant effusions were tested. Cells isolated from the blood of patients with leukemic spread of lymphomas of low malignancy yielded a weak staining comparable to that of normal mesothelial cells from non-tumorous cavity fluids. The detection of estrogen and progesterone receptors (ER and PR) localized in the cell nucleus can be achieved by the same assay. The reaction is enhanced by incubation of the tumor cells for 30 min at 37 degrees C prior to fixation. Pleural effusions from 20 patients with breast cancer were tested. ER was positive in 13 and PR was positive in 12 of the 20 samples. In 5 cases there was a divergent reaction with ER and PR antibody. The hormone receptors of the primary tumor were known in 15 (ER) and 14 (PR) patients, respectively. In each cohort there was only one case with a negative reaction of the primary tumor and a positive reaction with the isolated tumor cells from the pleural effusions. These results indicate that the demonstration of hormone receptor proteins in cells from malignant effusions is possible and that there is a correlation with the status of the primary site of cancer.
Breast Neoplasms↗
Serum antibodies to a normal cellular protein (P-65) in patients with Hodgkin's disease.
Samples from the L-428 Hodgkin's cell line and from several other lymphoma cell lines were lysed and the soluble proteins were subjected to one- or two-dimensional gel electrophoresis. The separated proteins were transferred to nitrocellulose by the Western Blot technique; antibody reactivity was detected by an immunoperoxidase reaction. Of 152 sera from patients with Hodgkin's disease (HD) 26 (17%) reacted with protein P-65 whereas in the control group, consisting of 35 healthy persons and 20 patients with other malignant diseases only 1 serum was reactive. Thus, the difference between the two groups was highly significant (P less than 0.01). These antibodies were most common in stage III HD. Splenectomy had no effect on the incidence of these antibodies, and there seemed to be no correlation with B-symptoms or with the histological subtype.
Adult↗
[Effectiveness and side effects of recombinant alpha-2a interferon in patients with metastatic hypernephroma].
Eighteen patients with advanced renal cell carcinoma were treated with recombinant interferon alpha-2a (18 million IU s.c. three times a week). 9 patients had received prior hormonal therapy (tamoxifen). On entering the therapeutic schedule with recombinant interferon alpha-2a, all patients had progressive disease with multiple metastases. Flu-like symptoms occurred in general; they were severe in 9 cases. An impairment of the liver was shown by a rise of SGOT/SGPT and alkaline phosphatase in 2 patients, the gamma-GT was raised in 7 patients. The kidney function worsened in 3 cases. A fall in blood cell counts was seen in 8 cases. Five patients developed a transient stomatitis. Five patients (5/18) showed stable disease with the longest response duration of 17 months. One patient showed a complete remission 5 months after starting therapy for a duration of 7 months until now. Both the limited benefit from this treatment as well as some severe side effects indicate the need for further special evaluation and possible improvements of this therapeutic approach.
Adult↗
Characterization of the Epstein-Barr virus-induced early polypeptide complex p50/58 EA-D using rabbit antisera, a monoclonal antibody, and human antibodies.
A polypeptide complex (p52) belonging to the D-subspecificity of the EBV-induced early antigens (EA-D) was purified from chemically induced P3HR-1 cells. Rabbit antisera raised against the isolated polypeptides reacted with components of EA-D as could be shown by indirect immunofluorescence and immunoperoxidase staining of IdU-induced EA positive Raji cells, ELISA, and immunoblotting. In one-dimensional immunoblots the rabbit antisera detected a predominant polypeptide complex of 52 kDa. Two-dimensional immunoblots prepared with proteins from IdU-induced Raji cells showed that the rabbit sera detect three series of polypeptides of 52 kDa (pl 8.5-6.2), 55-58 kDa (pl 6.2-4.5), and 48-50 kDa (pl 6.0-4.5). These three groups of polypeptides could also be identified by 50 high titered anti-EA-D positive human sera and a specific monoclonal antibody (R3) as being the main components of EA-D in Raji and B95-8 cells. All polypeptides of the p50/58 complex showed DNA binding properties either by themselves or by an interaction with other proteins. When TPA or IdU-induced Raji cells were labeled with 2Pi, two phosphorylated polypeptides pp50 and pp58 could be immunoprecipitated with the rabbit sera and a high anti-EA titered human serum. The time course of the synthesis of polypeptides associated with the EA-D complex was studied by 2-D immunoblots: EA polypeptides of 52 kDa appeared as early as 6 hr after the addition of IdU to Raji cells in culture, polypeptides of 55-58 and 48-50 kDa after 18 and 25 hr, respectively. The coordinated appearance of these groups of polypeptides and their similar size and reactivity with human sera and rabbit antisera produced against the isolated p52 as well as with a monoclonal antibody (R3) suggested that most of these polypeptides are derived from post-translational modifications of one or a few initially synthesized polypeptides, possibly p52. Phosphorylation seems to be at least one possibility of post-translational modification.
Animals↗
[Infection of the CNS caused by Listeria monocytogenes].
This report examines two cases of infection of the central nervous system by Listeria monocytogenes (L.m.). Both cases show that listeriosis is not only a differential diagnosis of purulent meningitis, but can also be the cause of an isolated brain stem syndrome with normal cerebrospinal fluid cell count. The prognosis depends crucially on the early antibiotic therapy (ampicillin). The first patient was a chronic alcoholic. He died of fulminant septic shock and meningitis with brain stem encephalitis (cell count of cerebrospinal fluid: 10500/microliters). L.m. was isolated from blood cultures and from cerebrospinal fluid. The second patient had no indications of preexisting immunological disorder. Two days after perianal injections for haemorrhoids, symptoms of a progredient brain stem syndrome developed. The cell count of cerebrospinal fluid was only 10/mu, but L.m. was isolated from blood cultures. The patient died of circulatory failure. At autopsy, a brain stem encephalitis and cerebellitis with inflammation of the surrounding leptomeninx was identified.
Cerebral Ventricles↗
Increased incidence of IgA antibodies to the Epstein-Barr virus-associated viral capsid antigen and early antigens in patients with chronic lymphocytic leukemia.
Antibody titers to Epstein-Barr virus (EBV)-associated early antigens (EA) and the viral capsid antigen (VCA) were determined by ELISA on 263 sera obtained from healthy donors, patients with Hodgkin's disease (HD), non-Hodgkin lymphomas (NHL), infectious mononucleosis (IM), Burkitt's lymphoma (BL), and nasopharyngeal carcinoma (NPC). As expected, most lymphoma patients showed markedly elevated anti-VCA IgG and anti-EA IgG antibody titers. Only one patient in the NHL group (n = 56) consisting of patients with lymphomas other than chronic lymphocytic leukemia (CLL) and hairy-cell leukemia (HCL), and 3 patients with HCL (n = 19) had high antibody titers of the IgA class to VCA and EA. Seventeen out of 48 patients (36%) with CLL had high IgA anti-VCA titers and 10 of these sera (21%) also contained IgA anti-EA. The geometric mean titer (GMT) of IgA anti-VCA was 2,510, the GMT of IgA anti-EA was 780. These antibody titers were about 10 times lower than the corresponding GMT of the NPC patients investigated in this study. The elevated IgG and IgA antibody titers to VCA and EA in CLL and HCL patients seem to reflect an immunodeficiency secondary to the malignant disease leading to reactivation of latent EBV infection. The possibility that at least some of these B-cell lymphomas are associated with EBV cannot be excluded.
Adult↗
Purification of a polypeptide complex (p52) belonging to the D-subspecificites of Epstein-Barr virus-induced early antigens.
A two-dimensional immunoblot analysis of chemically induced EBV DNA carrying Burkitt's lymphoma cell lines shows besides a large number of minor components at least two major groups of polypeptides: the most prominent group of polypeptides is observed in the range of 48 to 58 kDa (pI 4.5 to 8.5) and another group at 38/36 kDa (pI 4.4). A polypeptide complex (p52) belonging to the Epstein-Barr virus (EBV)-induced early antigen complex (EA) has been isolated from IdU-induced Raji and B95-8 cells as well as from n-butyrate-induced P3HR-1 cells. The p52 polypeptides have been purified by chromatography on Blue-, DEAE-, CM-, and Phenyl-Sepharose. The purification of these components of the EA complex was monitored by ELISA and by two-dimensional immunoblots using a well-characterized high anti-EBV positive human serum. The isolated polypeptides have an apparent mol wt of about 52,000 Da as determined under nondenaturing conditions by gel filtration chromatography on Sephacryl S-300. One- and two-dimensional immunoblots show a major group of polypeptides of 52 kDa (pI 8.5 to 5.5) with EA activity and some minor components with smaller size up to 40 kDa. The latter seem to be generated by limited proteolysis of p52 polypeptides. The EA activity of the isolated polypeptides could be confirmed by their reaction with IgG anti-EA positive as well as IgA anti-EA positive sera by ELISA. The purified polypeptide complex did not react with anti-EA-D negative, anti-EA-R positive sera obtained from patients with African Burkitt's lymphoma, suggesting that these polypeptides belong to the EA-D complex. The monoclonal antibody R3 reacted with the isolated 52 kDa components of EA suggesting a common epitope present on these polypeptides, the same result was obtained with three rabbit sera produced against the isolated polypeptide complex.
Antibodies, Viral↗
[Primary orthostatic hypotension].
We studied a 25-year-old man suffering from primary orthostatic hypotension whose blood pressure decreased to 65/45 mm Hg during orthostasis and to 95/70 mm Hg during ergometric exercise (50 and 100 watt), and whose heart rate responses were inadequate. Resting catecholamine levels were within the normal range and did not show any significant increase related to orthostasis or to ergometric exercise. Hypersensitivity was observed to low doses of intravenous noradrenaline and isoproterenol. Specific binding of 3H-Yohimbine to intact platelets revealed a normal number of alpha-2-adrenoreceptors in agreement with the adrenaline-induced platelet aggregation in vitro, which was, however, in contrast to hypersensitivity to noradrenaline. Specific 3H-Dihydroalprenolol binding to intact polymorphonuclear leucocytes revealed an increased beta-2-adrenoreceptor density in agreement with hypersensitivity to Isoproterenol. Prescription of Fludrocortison improved orthostatic hypotension.
Adult↗
[Demonstration of IgG- and IgA-antibodies against Epstein-Barr virus-associated antigens with a microtiter enzyme immunoassay system. The determination of serum antibodies against the viral capsid antigen and the early antigen complex in the sera of tumor and infectious mononucleosis patients].
An enzyme-linked immunosorbent assay (ELISA) using micro-titre plates has been established for determination of IgG and IgA antibodies to the Epstein-Barr virus (EBV)-associated early antigen complex (EA) and the viral capsid antigen (VCA). The results obtained correlated well with the antibody titre determinations by the standard immunofluorescence technique. The ELISA was found to be about 15 to 30 times more sensitive than the immunofluorescence assay. Because of its high sensitivity, specificity and rapid performance the enzyme-immunoassay is a useful tool for large scale epidemiological studies of EBV-associated diseases as well as for the early diagnosis and monitoring of patients with EBV-associated tumours: Burkitt's lymphoma and nasopharyngeal carcinoma. IgG and IgA antibody titres to EA and VCA were determined in sera obtained from healthy controls, patients with EBV-associated tumours, infectious mononucleosis and various diseases known to be associated with EBV-reactivation, i.e. lymphomas and leukaemias. Except in sera of patients with nasopharyngeal carcinoma, IgA antibodies to VCA and EA could only be detected at a rather high frequency (36% IgA anti-VCA+, 21% IgA anti-EA+) and in substantial titres in sera obtained from patients with chronic lymphatic leukaemia.
Antibodies, Viral↗
Immunoglobulin light chains in chronic lymphocytic leukemia and related diseases.
The combination of two-dimensional mini gel electrophoresis with the 'western blot' technique proves to be a powerful tool in characterizing lymphoid cells. By testing for kappa and lambda immunoglobulin light chains we were able to differentiate between mono-, oligo-, and polyclonal B-cell diseases. The distribution of the lambda isotypes of 24 cases with chronic lymphocytic B-cell leukemia (B-CLL) tested was not random when compared to the distribution of the lambda light chains in B-lymphocytes of normal persons. This might implicate a genetic link between the lambda loci (chromosome 22) and the development of the lambda-CLL.
Adult↗
Human adult T-cell leukaemia virus is distinct from a similar isolate of Japanese monkeys.
We have compared the structural polypeptides of an adult T-cell leukaemia (ATL) virus (ATLV) isolate from a Japanese patient with ATL with those of a similar virus derived from a Japanese macaque monkey. Both are distinct but related entities. Their core polypeptides p19 could not be distinguished, but p24, another core polypeptide, and their envelope glycopolypeptides differ. The human virus directs the synthesis of a single intracellular glycopolypeptide, gp68, while the macaque virus specifies two such glycopolypeptides, gp57 and gp50. Furthermore, the glycopolypeptides of both viruses are serologically distinct. Thus, these viruses represent subtypes of the ATLV family and the macaque virus is apparently not involved in human ATL.
Animals↗
Purification of a protein (60K/58K) associated with the Epstein-Barr virus-induced early antigen complex in Raji cells.
A double antibody sandwich ELISA has been established for the detection and quantitation of EBV-associated early antigens (EA) in IUdR-induced Raji cells. The EA complex extracted from Raji cells could be separated by ion exchange chromatography and isoelectric focusing into several components. One EA-associated subspecificity has been purified by DEAE-, CM-, and Blue-Sepharose chromatography followed by isoelectric focusing. The isolated protein has an apparent molecular weight of 240,000 +/- 20,000 daltons under non-dissociating conditions on Sephacryl S-300, an isoelectric point of 4.5, and seems to be composed of two polypeptides of 60,000 and 58,000 daltons as shown by SDS-gel electrophoresis and two-dimensional gel electrophoresis. Preliminary data indicate that the 58,000 polypeptide is generated by limited proteolysis of the 60,000 polypeptide. The EA activity of the isolated protein has been confirmed by the double antibody sandwich ELISA and its reactivity with anti-EA-positive sera in an ELISA for the detection of anti-EA antibodies.
Antibodies, Viral↗
Characterization of lymphoid cells by two-dimensional mini gel electrophoresis of proteins.
Cytosol proteins from normal lymphocytes, leukemic lymphocytes, and different cultured lymphoid cell lines were separated by two-dimensional mini gel electrophoresis. By staining with Coomassie blue, specific protein patterns were obtained. Very similar gel maps were produced by the cytosol proteins of chronic lymphocytic leukemia cells, hairy cells, and of in vitro grown B cells. Protein 36/6.2 (molecular weight/isoelectric point), consistently present in these cells, could not be demonstrated in normal lymphocytes. For the comparison of control Raji cells--an Epstein-Barr-Virus (EBV)-DNA carrying Burkitt's lymphoma cell line--with Raji cells induced for early antigen (EA) production, 35S-methionine-labelled total cell lysates were analyzed. In the induced cells, an additional protein (100/5.5) was found; this might be one of the immunologically defined EBV-associated antigens. These results demonstrate that two-dimensional mini gel electrophoresis can be useful for the characterization of leukemic cells in addition to the morphological, cytochemical, and surface marker analyses.
Adult↗
Simplified method for the discrimination of acute myelogenous and non-myelogenous leukemias.
Rapid discrimination of acute myelogenous and non-myelogenous leukemias is of great importance when chemotherapy is urgently needed in severely ill patients. For decades the most reliable cytochemical method for this classification is the demonstration of myeloperoxidase in blast cells [1-4, 6, 7]. We combined the simplified myeloperoxidase stain as described by Kaplow with a brief stain similar to Pappenheim's procedure or with commercially available Hemacolor rapid blood smear: this proved to be a simple staining method that permits good morphological judgment of the cells as well as reliable demonstration of peroxidase activity. This procedure takes less than 10 min using Hemacolor and can easily be done with prepared solutions without technical assistance.
Cell Nucleus↗
The influence of the calcium/phosphorus ratio in the diet on human bone disease, the role of nutritive secondary hyperparathyroidism.
60 out of 100 patients suffering from severe periodontal disease received a daily calcium supplementation of 1,000 or 2,000 mg during a period of 9 or 12 months. Different investigations such as PTH and serum parameters were not influenced by this treatment. However, animal experiments, as well as acute studies in men involving an oral phosphorus load, suggest that parathyroid secretion might be increased by a lowered calcium/phosphorus ratio. Therefore it is possible that a diet too low in calcium could contribute to the pathogenesis of periodontal disease. The lacking effect of calcium therapy implies very strongly that other factors are more important for the development of this disorder.
Calcium↗
Enrichment of special Novikoff hepatoma and regenerating liver mRNA by hybridization to cDNA-cellulose.
Total polysomal poly(A)+ RNA of Novikoff hepatoma and of 18-hr regenerating rat liver were compared by analysis of their in vitro translational products on two-dimensional isofocusing/sodium dodecyl sulfate gels. This technique resolved the translated proteins sufficiently to permit detection of quantitative and some qualitative differences between the two mRNA populations. Excess cDNA from regenerating liver or Novikoff hepatoma, covalently linked to cellulose, was used to adsorb the complementary mRNA sequences from Novikoff hepatoma or regenerating liver. As shown by two-dimensional gel electrophoresis, the translated products of the bound mRNA fractions contained proteins common to both tissues. Novikoff hepatoma mRNA which did not bind to regenerating liver cDNA was enriched in sequences encoding for proteins 11/5.1, 15/6.8, 40/8.2, and 65/5.1 (shown as molecular weight/pI). These polypeptides were not detectable in the translational products of regenerating liver mRNA. Regenerating liver mRNA that was not bound to Novikoff hepatoma cDNA was enriched in sequences coding for proteins 12.5/4.9, 13.5/7.4, 17/8.2, 24/5.5, and 46/6.4; these proteins were not found in the translational pattern from Novikoff hepatoma. These results show that adsorption of mRNA to solid-phase cDNA provides a valuable technique for differentiating mRNA species in related tissues and for corresponding enrichment of these specific mRNAs.
Animals↗