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Herpes simplex virus type 2 (HSV-2) is the main cause of herpes genitalis, a recurrent sexually transmitted disease. By the use of routine Serologie methods (complement fixation test, enzyme immunoassay), virus carriers are difficult to identify because of strong antibody cross reactions with antigens of HSV-1 which is ubiquitously spread throughout the population. We introduce a microtechnique Western blot system loaded with HSV-1 and HSV-2 type-specific and common antigens on separated nitrocellulose strips. By the simultaneous evaluation of Immunologie reactions with both strips, the occurrence of HSV-2 specific antibodies can be sensitively detected in serum specimens containing antibodies to HSV-1. A total of 158 serum specimens were analyzed and the results obtained by Western blot were compared to those of a screening ELISA and virus isolation performed with smears of herpes lesions.
An agreement of 97.9 % was assessed between Western blot and virus isolation to detect an HSV-1 and HSV-2 infection. Less specific serologic results were produced by the screening ELISA on HSV-2 antibodies which correlated in 85.4 % (41/48) with virus isolation and typing. Concerning HSV-2 antibody testing, Western blot and ELISA showed an overall agreement in 89.8 % of the sera investigated.
As shown by our data, the HSV type specific Western blot proved to be a specific, reproducible and standardized technique. It can be utilized for both sero-epidemiological surveys and determination of the HSV immune status.
Total body water increases in pregnancy and while the uterus, placenta, fetus, and amniotic fluid constitute part of this increase, the largest component is in the extracellular water. Fat stores also increase and thus the distribution volumes of all drugs expand, but the major effect is seen in polar drugs which are confined to the extracellular space. Cardiac output and renal function also increase and elimination of polar drugs is acelerated. In contrast, the elimination of lipophilic drugs may be retarded, and the effect on intermediate drugs is variable. Polar drugs cross the placenta slowly and accumulate in amniotic fluid and therefore in the fetal gut lumen. Lipophilic drugs cross the placenta rapidly and their transplacental distribution is dependent on relative maternal and fetal affinity: this is determined largely by protein binding on either side of the placenta. The. fetus and neonate dispose of all drugs less rapidly than adults, the most efficient elimination processes being sulphate conjugation and renal excretion.
(±)-Aeroplysinin-1, an optically active 1.2-dihydroarene-1.2-diol. was isolated from the marine sponges Verongia aerophoba (+-isomer) and lanthella ardis (--isomer). For the experiments presented we used the +-isomer from Verongia aerophoba. Here we describe the hitherto unknown biological and pharmacological property of this compound to display pronounced anticancer activity against L5178y mouse lymphoma cells (ED50: 0.5 μm). Friend erythroleukemia cells (ED50: 0.7μm) , human mamma carcinoma cells (ED50: 0.3μm) and human colon carcinoma cells (ED50: 3.0 μm) in vitro. Furthermore, aeroplysinin caused a preferential inhibition of [3H]thymidine (dThd) incorporation rates in L5178y mouse lymphoma cells if compared with murine spleen lymphocytes in vitro. At concentrations between 1.1 and 28.5 μm, the [3H]dThd incorporation rates in L5178y cells were suppressed to 28% -0% but only to 78% -18% in murine spleen lymphocytes. The same differential effect in vitro was found with the following epithelial cells: 14.70 μm of the compound were required to inhibit normal human fibroblasts to 50% , but only 2.9 μm in the assays with human malign keratinocytes or malignant melanoma cells to observe the same inhibitory effect. Moreover, aeroplysinin-1 displayed antileukemic activity in vivo using the L5178y cell/NMRI mouse system; administered at a dose of 50 mg/kg for five consecutive days, the T/C (% ) value was determined to be 338. Preliminary toxicology studies revealed an acute LD50 of 202 mg/kg and a subacute LD50 of 150 mg/kg. Aeroplysinin-1 is neither a direct mutagen nor a premutagen in the umu/Salmonella typhimurium test system.
Membrane-Phloretin Interaction, Infrared Raman, ESR Spectroscopy The transport inhibitor phloretin was bound to human red cell membrane and the concomitant structural changes were observed by spectroscopic methods. By the spin labeling method a decrease in fluidity of the membrane was found at 1 and 10 |iM concentrations of the reagent. This result was obtained with the 2-(3-Carboxypropyl)-4,4-dimethyl-2-tridecyl-3-oxazolidinyloxyl, and the 2-(14-Carboxytetradecyl)-2-ethyl-4,4-dimethyl-3-oxazolidinyloxyl lipid spin labels. Infrared spectroscopy of modified membranes revealed an intensity increase of the POO~ band at about 1250 cm-1. Moreover, a shift of the peak at 1050 cm -1 to 1100 cm-1 was observed in the presence of phloretin. Raman spectroscopy of the membranes did not contradict the results found with infrared and ESR spectroscopy: In the phloretin modified membrane we observed a lack of the band at 1085 cm-1, which leads to suggest that the POO" and/or C-C regions are less fluid. Changes of the extracted red cell membrane lipids were less characteristic, and the results differed from those found in red cell membrane.
1-anilino-naphthalene-8-sulfonate (ANS) fluorescence measurements have revealed that red blood cell membrane of the Rhnull type undergoes a transition at about 16 degrees C. In contrast, viscosity measurements of the extracted membrane lipids showed the usually observed transition at about 18 degrees C. Lower values of titratable sulfhydryl (SH) groups were observed in Rhnull membrane using 5,5'-dithiobis-(2-nitro-benzoic-acid) (Nbs2). In contrast, disulfide bonds in Rhnull membrane were estimated to be about 3 times the value of the controls. Spin labeling experiments using 2-(3-carboxypropyl)-4, 4 dimethyl-2-tridecyl 3-oxazolidinyloxyl were carried out with phospholipase A2 modified membranes. The mobile part of the spectra was significantly increased on the Rhnull membrane. In the presence of D-glucose, infrared spectrometry showed a larger reduction of the intensity of the POO-band in Rhnull membrane. In contrast to controls, binding of the reagent diethylpyrocarbonate resulted in no significant changes of the Rhnull membrane as determined by electron spin resonance (ESR) measurements. D-glucose transport activity was found to be at the upper level of a group of Rh positive and Rh negative persons. It is suggested that the intensity of the polar protein-lipid interaction is reduced in Rhnull membrane.
Calcification, Collagen Membrane, Ca/P Ratio Dependence Spontaneous calcification of a membrane made of native collagen has been investigated. The method permits independent variation of calcium and phosphate concentrations. With increasing phosphate concentration the precipitation of calcium-phosphate on the collogen occurs at a conspicuously lower calcium concentration as with a number of other membranes.
The norepinephrine content of adipose tissue is shown to be very different in various animal species and different sites of origin, ranging from 0.03-1.4 μg/g. Adipose tissue also contains considerable amounts of serotonin (0.01-1.04 μg/g) and histamine (0.1-13.6 μ/g). Changes in the norepinephrine content of adipose tissue after the injection of either reserpine analogues or monoamine oxidase inhibitors followed a pattern similar to that found in the heart and brain, indicating that the storage mechanism in these organs is basically the same. In contrast to norepinephrine, serotonin in adipose tissue is rather resistant toward depletion by reserpine. Adipose tissue also contains monoamine oxidase and catechol-O-methyl-transferase activity, which are usually highest in tissues also rich in norepinephrine.