The interrelationship between Coxsackie B4 (CB4) virus antibodies, islet-cell antibodies and HLA B8 was studied in 67 patients with juvenile-onset diabetes mellitus (JOD). In 41 JOD patients (61%) CB4 antibodies (greater than 1:4) were found. A high prevalence (69%) of significant CB4 antibody titers (greater than 1:64) was observed in patients with age at onset between 11 and 20 years compared with those with age at onset below or over that age-range (27% and 33%, respectively). Islet-cell antibodies (ICA) could be detected only in 11 JOD patients, which is probably due to the relatively long mean duration of disease. However, 8 of them presented with significant CB4 antibody production. Although no correlation between CB4 antibodies and the JOD-associated HLA-antigens (B8, Bw15, Cw3 and B7) could be detected in the entire group of JOD patients, a close association between ICA persistence and HLA B8 was found. Furthermore, in the ICA-positive, moderate or high CB4 antibody titers were prevalent. Thus, the present data point to a possible interrelationship between CB4 virus antibodies, persistent ICA production and HLA B8 in a minor proportion of JOD patients. However, it seems premature to assume direct implication of these factors in the pathogenesis of JOD.

Antibodies, Viral↗

PubMedDate not supplied

Antibody-dependent killer-cell function in insulin-dependent diabetes mellitus.

Antibody-dependent killer-cell activity (ADCC) was determined in 36 insulin-dependent diabetics (IDD) and 32 controls. The medians of cytotoxic indices obtained by using either antibody coated chicken red blood or HeLa cells as targets were statistically significantly reduced in the diabetics (P less than 0.05 for both systems). However, due to the wide range and considerable overlap of the cytotoxic indices observed in patients and controls, the biological significance of these mathematical differences remains to be determined. The duration of the disease did not have any influence on killer (K) cell function and only a slight tendency for decreased ADCC during episodes of poor metabolic control was noted. Further analysis of the influence of diabetes-associated factors did not reveal any definite correlation between the functional K cell deficit, the insulin dosage administered, the insulin antibody titers, the blood glucose at the time of sampling, the 24 hour glucosuria and IDD-associated immunogenetic factors.

Adult↗

PubMedDate not supplied

Light microscope and immunohistological investigations on the brain of Borna disease virus-infected rabbits.

We studied the distribution and composition of perivascular cellular infiltrates and the distribution of the virus-specific antigen of Borna disease in the brains of rabbits inoculated either intracerebrally or intraperitoneally with infectious virus suspensions. Using standard histopathological and immunofluorescence techniques, there was, in all animals, a meningeal inflammatory reaction together with perivenous cellular infiltrates which were most numerous in the structures of the limbic system and basal ganglia, but also prominent in the spinal ganglia and area postrema. Studies with fluorescent anti-IgG antibody (direct technique) showed IgG in 20-25% of the cells in any one perivascular cuff, suggesting that most of the infiltrating cells were T-cells and that B-cells were fewer. Studies with virus-specific antibody showed that, in the early stages of the infection, there was evidence of a globular antigen mainly in neurons, while, at later stages, virus antigen was also demonstrated in oligodendrocytes and microgliocytes. The route of inoculation of the virus had no determining effect on the character or distribution of the cellular infiltrates or on the distribution of virus-specific antigen in the nervous system.

Animals↗

PubMedDate not supplied

Long term cultures of neural retina and pigment epithelium from newborn rabbits.

In vitro cultures of neural retina, obtained after dispersion and trypsinization of tissue fragments, were composed of 3 morphologically distinct types of neural cells, as demonstrated by silver impregnation. They resembled ganglion or receptor cells, horizontal or amacrine cells, and bipolar cells of the intact retina. Pigment epithelium was cultured without trypsinization. Both kinds of techniques may prove helpful for long term experiments in neurobiology and neurovirology.

Animals↗

PubMedDate not supplied

Optimization of fast-fluorescence in situ hybridization with repetitive alpha-satellite probes.

A rapid FISH (fluorescence in situ hybridization) technique (Fast-FISH) for quantitative microscopy has been recently introduced. For highly repetitive DNA probes the hybridization (renaturation) time and the number of necessary washing steps were reduced considerably by omitting formamide or equivalent denaturing chemical agents. Due to low stringency conditions major and minor binding sites of the probes used showed visible FISH signals well suited for quantitative image-microscopy. The discrimination of minor and major binding sites was possible by automated image-processing. Here, a further, quantitative optimization of the Fast-FISH technique is described that allows to clearly discriminate major and minor binding sites of alpha-satellite probes by an easy image classification parameter. With respect to the optimization it was necessary to verify two sensitive parameters (hybridization time and temperature) of the given rapid FISH protocol. As examples the systematic optimization for the two probes D12Z2 (major binding site on the centromere of chromosome 12) and D8Z2 (major binding site on the centromere of chromosome 8) are shown. The optimal hybridization conditions concerning rapidness and quality of chromosome morphology were obtained using a hybridization temperature of 70 degrees C and a hybridization time of 60 min. For these conditions major and minor binding sites were clearly discriminated by the intensity maximum Smax of the corresponding FISH-spots.

Binding Sites↗

PubMedDate not supplied

Painting of human chromosome 8 in fifteen minutes.

The technique of chromosome-in-situ suppression (CISS)-hybridization (chromosome painting) has now been well established. However, all standard protocols so far require long renaturation times (typically 12 hours and more). Here, we describe a new, extremely fast protocol for chromosome painting using a commercially available, directly fluorescence labelled probe for chromosome 8. The hybridization conditions used omit separate preannealing procedures and denaturing chemical agents. The renaturation time required for chromosome painting was reduced to 15 minutes. In addition, most washing steps were eliminated. As a consequence, the entire painting procedure was feasible in less than half an hour.

Chromosomes, Human, Pair 8↗

PubMedDate not supplied

Immediate-type allergy against insulin itself: clinical and immunologic studies on a diabetic patient with insulin intolerance.

In this report clinical and immunologic data of a female diabetic patient with immediate-type allergy against insulin preparations of different species, including pancreatic human insulin, biosynthetic human insulin and semisynthetic human insulin are described. The determination of insulin-specific antibodies showed relatively high IgE-antibody titers in contrast to low IgG-antibody concentrations, which is in accordance with the assumption that the ratio of IgE- to IgG-insulin antibodies is important in the outcome of clinically manifest allergic reactions. Negative results for cutaneous delayed-type insulin hypersensitivity and for stimulation of lymphocyte blastogenesis exclude type IV immunopathogenic reaction in this insulin-allergic patient. Preliminary data suggest that inherited individual factors--such as DR2, DR3 positivity in this diabetic individual--might predispose diabetic patients to insulin allergy.

Aged↗

PubMedDate not supplied

Dose escalation of ara-c may improve response rates in a subgroup of chronic myeloid leukemia patients with poor response to interferon-alpha and low-dose ara-C.

The present analysis was performed to evaluate the impact of cytosine arabinoside (ara-C) dose escalation on hematological and cytogenetic responses in patients with chronic myelogenous leukemia (CML) who failed to respond to low-dose ara-C (LD ara-C) at a dose of 10 mg/m2/d over 10 days per month and interferon-alpha (IFNalpha, 3.5 MU/d). Following the same administration schedule, dose escalation of ara-C to 15 and 20 mg/m2/d 1-10 was performed in 36 of 119 patients (30%) due to inadequate hematological response and/or disease progression. As a result, improvement of hematological and cytogenetic responses was achieved in 22 (61%) and nine (25%) patients, respectively. Escalated ara-C dose levels were usually well tolerated, although some patients experienced deterioration of preexisting side effects. Our results support the critical role of ara-C dose towards a better disease control in CML.

Adolescent↗

PubMedDate not supplied

Collagen antibodies and collagen- anticollagen immune complexes in rheumatoid arthritis.

Serum, synovial fluid and synovial fluid cells of 14 patients with classic rheumatoid arthritis and of 5 controls were investigated serologically in regard to rheumatoid factors and collagen antibodies and by immunofluorescence double staining in regard to localization of collagen and gammaglobulin in fluid cells. Three patterns of staining were observed: a) gammaglobulin inclusions, b) gammaglobulin and collagen in identical localization, c) collagen inclusions. While distinct staining for gammaglobulin appeared only in seropositive cases, collagen inclusions were only observed in RA, however in seropositive and seronegative cases. In 5 RA cases collagen and gammaglobulin were demonstrated additionally in such identical localization that conclusion in regard to collagen-anticollagen immune complexes seems justified. 9 of 14 RA cases had collagen antibodies, all cases were collagen antibody positive which had collagen and gammaglobulin in identical localization in same inclusions.

Antigen-Antibody Complex↗

PubMedDate not supplied

'In situ' high pressure confocal Ca(2+)-fluorescence microscopy in skeletal muscle: a new method to study pressure limits in mammalian cells.

We combined 'in situ' high pressure microscopy with confocal laser scanning microscopy to directly study Ca2+ homeostasis in intact mammalian (murine) skeletal muscle fibres during high pressure exposure up to 35 MPa. Cytosolic Fluo-4 and mitochondrial Rhod-2 Ca2+ fluorescence were simultaneously monitored. To separate changes in Ca2+ and direct/indirect effects of pressure on the dye, experiments in permeabilized ('skinned') muscle fibres were performed at a fixed Ca2+ concentration. Normalized Fluo-4 fluorescence sharply declined up to 10 MPa but showed a plateau between 10 MPa and -35 MPa. In the intact fibre, Fluo-4 fluorescence exponentially decreased during pressurization to 35 MPa with a pressure constant of pi-5 MPa whereas mitochondrial Rhod-2 fluorescence exponentially increased with a four-fold larger pi. Holding the pressure at 35 MPa almost did not change Fluo-4 fluorescence. However, Rhod-2 fluorescence started to decrease after -40 min. Upon decompression, Rhod-2 and Fluo-4 fluorescence increased exponentially with similar pi. However, initial Fluo-4 fluorescence values were not restored. Our results are in agreement with pressure induced Ca2+ leakage from the sarcoplasmic reticulum. Ca2+ might then be taken up in large amounts by mitochondria preventing cytosolic increase in Ca2+. Prolonged pressure applications (-40 min at 35 MPa) seem to destabilize mitochondrial function with release of Ca2+ from mitochondria back into the cytosol and eventually mechanical activation resulting in irreversible contractures. The pressure induced disturbance of Ca2+ homeostasis might have important implications for the pressure exposure limits and/or dive profiles of deep sea mammals.

Aniline Compounds↗

PubMedDate not supplied

[Effect of low molecular dextran on maternal microrheologic parameters, fetal heart rate and transcutaneous fetal oxygen partial pressure in labor].

The results from animal experimental research have shown that the infusion of dextran 40 increased the uterine blood flow mainly by decreasing the vascular resistance and by this improving the oxygen supply to the fetus. Dextran 40 given in a dosage of 50 ml/min within 10 minutes to 11 pregnant women during parturition showed that the fetal blood PO2 increased if the PO2 before infusion was low and decreased if the PO2 before infusion was high. Therefore the question enhanced if the reduced maternal oxygen capacity results in a reduction of fetal PO2 (tcPo2) too. After informed consent 10 healthy pregnant women at term with a normal pregnancy got an infusion of 500 ml dextran 40 during 30 minutes. The following maternal parameters were measured: heart rate, mean arterial blood pressure, intrauterine pressure and transcutaneous carbon dioxide tension (tcPCO2). Maternal microrheological parameters before and after infusion of dextran 40 were estimated as following: plasma viscosity, erythrocyte aggregation, serum osmolality, colloid osmotic pressure, hemoglobin and haematocrit. The mothers were laying in the side position and their ventilation was controlled by tcPco2 (Dräger, Lübeck). The following fetal parameters were registered: basal fetal heart rate, transcutaneous fetal oxygen partial pressure (tcPo2) and the umbilical cord blood gases. After rupture of the fetal membranes and at a cervix dilatation of about 5-7 cm a tcPo2 electrode (Dräger, Lübeck) was fixed at the fetal scalp with an acrylate glue (Histoacryl) and by this fetal tcPo2 and its "flow" registered.(ABSTRACT TRUNCATED AT 250 WORDS)

Blood Pressure↗

PubMedDate not supplied

Experiences with continuous intraarterial administration of recombinant interferon alpha-2C (rIFN-alpha 2) for treatment of patients with advanced transitional cell bladder cancer.

The response of locally-advanced transitional cell carcinoma of the urinary bladder to intraarterial regional treatment with recombinant interferon alpha-2C (rIFN-alpha 2) was investigated. Patients received 4-weekly courses of i.a. continuous infusion of rIFN-alpha 2 (5 megaunits/24 hours administered into the a. iliaca interna via a subcutaneously implanted port-a-cath) alternating with 4 weeks i.m. treatment cycles (5 megaunits/day x 3 weekly). None of the 5 patients entered in the study showed evidence of objective tumor regression after a median of 4 i.a./i.m. treatment cycles. Constitutional, organ and local toxic effects were considerable even with that low-dosage schedule employed. The lack of efficacy and poor tolerance of the regimen suggest that i.a. administration of IFN is unlikely to yield an acceptable therapeutic index.

Aged↗

PubMedDate not supplied

Phase II trial of recombinant interferon alpha-2C in metastatic renal cell carcinoma.

A total of 18 patients with advanced metastatic renal cell cancer were treated with recombinant interferon alpha-2C (rIFN alpha-2C) at daily doses of 10 X 10(6) IU by intramuscular injection. All patients had evaluable metastatic lung, liver, or abdominal disease as measured by radiographic or computerized tomographic scans. In 2 of the 18 patients an objective response (1 CR, 1 PR) with a duration of +28 and 12 months, respectively was achieved. A 25$ to 50$ decrease in tumor measurements (MR) was seen in 2 additional patients; in 3 cases a stabilisation of the disease (SD) was observed, whereas it progressed in 11. 3/4 responding patients (including MR) and all 3 cases with SD had measurable disease in the lungs as predominant site of metastatic disease. Additional clinical characteristics of patients exhibiting response or SD to IFN therapy included prior nephrectomy, favourable initial performance status and limited metastatic disease. No serious haematologic or irreversible organ toxic effects were attributed to interferon. Several patients, however, had constitutional symptoms, and major dose reductions due to CNS toxicity became necessary in two. Further studies are warranted to evaluate the use of interferons in combination with cytotoxic drugs or other biologic response modifiers.

Adult↗