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Biomedical subjects

E D Wachsmuth

Publications and source records attributed to E D Wachsmuth.

At least 19 recordsLinked to original sources
PubMed1990

Course of murine leukemia retrovirus infection determined in vivo by magnetic resonance imaging.

Magnetic resonance imaging was applied to measure the volumes of spleens and lymph nodes of mice infected with three different leukemia retroviruses (LP-BM5 murine leukemia virus, Friend, and Rauscher) in vivo. Anesthesia by rapid intraperitoneal injection of Saffan was sufficient for magnetic resonance imaging and could be repeated at appropriate intervals. Eleven frontal magnetic resonance images through the abdomen with a center-to-center distance of 1.5 mm between adjacent slices were acquired simultaneously. To optimally demarcate spleens from the surrounding tissues, the magnetic resonance images were mildly T2-weighted for mice infected with LP-BM5 murine leukemia virus and mildly T1-weighted for those infected with Friend and Rauscher virus. Measurements requiring only 3 to 4 hours in groups of 24 to 28 mice were accomplished by using a standardized holder (i) accommodating two animals in the supine position and (ii) ensuring reproducible positioning in the magnetic resonance-instrument, and (iii) by reducing the number of phase-encoding steps of mildly T2-weighted magnetic resonance images from 256 to 128. Volumes of spleens and inguinal lymph nodes were calculated from the respective cross-sectional areas. The weights and magnetic resonance image-derived volumes of spleens and inguinal lymph nodes correlated well (r greater than 0.95). Despite large variations in the extent of splenomegaly and lymphadenopathy at any given time, the progression of the disease could easily be followed by repeating magnetic resonance imaging at intervals. Thus, statistically relevant results can be obtained in an infection model requiring the use of only a few animals.

Alfaxalone Alfadolone Mixture↗
PubMed1989

Stimulant effect of the immunomodulator MTP-PE on proliferation of monocytic cells in the guinea pig.

The effect of a single subcutaneous injection of various doses of the lipophilic muramyl tripeptide MTP-PE on cell proliferation was investigated in autoradiographs of histological sections of various organs of the guinea pig. The animals either received first MTP-PE in saline and then one 3H-thymidine pulse 1 h prior to sacrifice, or they were prelabeled with 3H-thymidine and then received MTP-PE. The number of proliferating cells increased up to between two and fivefold (marginally after 0.3 mg/kg and maximally after 30 mg/kg MTP-PE), but differed in the various organs. In addition, the time of the maximal increase varied between 5 h and 72 h after MTP-PE treatment and also depended on the organ. The majority of proliferating cells were of the monocyte lineage seen in conjunction with the vascular system. They were apparently promonocytes still capable of proliferation. Evidence for this conclusion is derived from (i) the distribution of 1 h-pulse-labeled cells in the various organ compartments in relation to the stimulated proliferation of the bone-marrow cells, and (ii) the distribution of the prelabeled, mainly bone-marrow derived cells, to the various organs. The augmented proliferation of the monocyte lineage is preceded by a dose-dependent, short-lasting increase in the proliferation of some epithelia and also by an increase in body temperature and a transient change in plasma proteins. These effects are part of a limited inflammatory reaction and may contribute to the immunostimulation.

Acetylmuramyl-Alanyl-Isoglutamine↗
PubMed1988

A comparison of the highly selective fluorescence staining of fungi in tissue sections with Uvitex 2B and Calcofluor White M2R.

The selective fluorescence staining of two fungi, Candida albicans and Blastomyces dermatitides, with Uvitex 2B and Calcofluor White M2R was studied in deparaffinized and frozen sections of mouse kidney and lung. Both fluorochromes emitted maximally at about 430 nm, independent of the mounting media (Kaiser's gelatin or Entellan). In addition to fungi, both fluorochromes also stained elastic fibres. The fluorescence intensity remained unchanged after storage of sections for more than 6 months in conventional slide boxes. The two fluorochromes showed the following differences: Calcofluor faded 1.25 times faster than Uvitex when illuminated with ultraviolet light. Calcofluor showed a greater affinity for tissues in general, and red cells and renal tubular casts in particular. Counterstaining of deparaffinized sections with Hemalum and Eosin reduced the fungi fluorescence and suppressed the general background fluorescence. However, it led to an intensification of Eosin staining and the fluorescence of red cells in Calcofluor-stained sections but not in Uvitex-stained ones. Similarly, the background fluorescence in frozen sections was reduced by Evans Blue, although elastic fibres still fluoresced after staining with Calcofluor. The degree of staining selectivity, and thus the contrast produced within a histological specimen, was greater with Uvitex 2B than with Calcofluor White M2R.

Animals↗
PubMed1988

Visualization of fungi in histological sections.

Deparaffinized kidney sections from mice infected with Candida albicans and lung sections from mice infected with Blastomyces dermatitides were stained with the stilbene derivative, Uvitex 2B (1%), and counterstained with haemalum and eosin. Fungi selectively stained with Uvitex 2B are visualized by blue fluorescence under incident illumination with ultraviolet light. Simultaneous or consecutive illumination with transmitted light permits the assignment of fluorescent fungi to haemalum-eosin-stained structures in the section. The most practical means of achieving a high optical contrast with Uvitex 2B in sections and good haemalum-eosin staining is to use the established haemalum-eosin technique, but with a solution containing both 1% eosin and 1% Uvitex 2B in place of eosin alone. Since Uvitex 2B stains all fungi investigated so far, it affords a simple, sensitive and inexpensive method of selectively detecting opportunistic fungal infections in conventional histopathology.

Animals↗
PubMed1988

Fully automated TV-image analysis of the cell-cycle: comparison of the PLM method with determinations of the percentage and the DNA content of labelled cells.

A cell-cycle analysis based on a fully automated TV-image scanning system is proposed to replace the laborious PLM method. To compare the efficiency of the two procedures, cell-cycle parameters were assessed in Ehrlich (diploid and hyperdiploid), L-1210, and JB-1 mouse ascites tumours and in rat jejunal crypts. The percentages of labelled mitoses (PLM) were counted visually on Feulgen-stained autoradiographs obtained at various times after a single 3H-thymidine pulse. The fraction of labelled cells (P) and the DNA ratio of labelled and unlabelled cells were measured by TV-image analysis in the same slides and plotted against time. Within practical limits, TV-image analysis using the P-curve gives the same results as the PLM method. Using the P-curve has the important advantage that its first part, beginning at the time of 3H-thymidine injection and ending at the first maximum, furnishes more information about the cell cycle than the corresponding part of the PLM curve. It can be used to compute tG2M tS and the ratio of the growth faction index to the cell-cycle time (IP/tC) whereas the first part of the PLM-curve reveals only the length of the S-phase (tS). The IP/tC ratio is a readily accessible measure of growth and increases when the cells divide more frequently. Cell death rates may be neglected since the ratio is determined within less than the duration of one cell cycle. Moreover, the data from the first part of the P curve indicate whether there is a large non-growth fraction. If the non-growth fraction is small, i.e. if IP approximately 1, the P curve need only be measured until the first maximum is reached so that fewer samples and animals are required. If the non-growth fraction is large or unknown, the cell-cycle parameters are calculated by reference to the position and size not only of the first minimum and the first maximum, but also of the second minimum of the P curve.

Animals↗
PubMed1988

Stimulation of cell proliferation in rabbits by MTP-PE, a lipophilic muramyl peptide.

The in vivo effect of the lipophilic muramyl peptide MTP-PE on the proliferation of blood cells and various tissues of the rabbit was studied by means of 3H-thymidine. Animals were killed up to 120 h after one or two i.v. injections of MTP-PE (10 mg/kg). MTP-PE caused a drastic effect on white blood cells: (1) neutropenia and lymphocytopenia occurring within 5 h was followed by leukocytosis of neutrophils and their juvenile forms by 24 h and thereafter, (2) within 24 h the number of prelabelled, i.e. recently regenerated, mononuclear cells in the bone marrow and the vascular system of various tissues increased approximately threefold, and (3) within 48 h the concentration of proliferating monocytic cells (1-h pulse labelling) rose to maximum levels of up to 20-fold in the lumina of blood vessels, particularly in capillaries of many organs. The number of proliferating cells also increased in the adventitia of medium and small arteries with a maximum at 48 h, whereas this occurred only later in the media and hardly at all in the intima. Thus, these proliferating, apparently monocytic cells are blood derived, and migrate into the tissue within 24 h after MTP-PE administration. In addition, proliferation in the epithelium of the bile ducts and oesophagus was also stimulated with a maximum at 24 h after MTP-PE. In contrast, enhanced proliferation occurred more slowly and to a lesser extent in hepatocytes, hepatic interstitial cells, and renal epithelial cells, consistent with a regenerative process after an inflammatory or toxic event.

Acetylmuramyl-Alanyl-Isoglutamine↗
PubMed1988

[Evaluation of the tolerance of the intra-ocular injection of hydroxypropyl methylcellulose in animal experiments].

Hydroxy-propyl-methyl-cellulose (HPMC) was injected into the anterior chambers or vitreous bodies of rabbit eyes in order to test local and systemic tolerance. In doing so the typical intraoperative complication of capsule rupture, which, in anterior segment surgery in man, allows HPMC to enter the vitreous body, was simulated. Neither clinically, nor in laboratory workup, nor histopathologically could any difference be shown between the local and systemic reactions of HPMC and those of DRSS. The data obtained in the study admit the conclusion that HPMC can be used in surgery of the anterior segment without any risk of adverse reactions.

Animals↗
PubMed1987

Re-examination of the effect of beta-adrenergic blocking agents on the proliferation of rat jejunal crypt cells using the stathmokinetic method.

Reports of the effects of beta-adrenergic receptor blocking agents on the proliferative activity of rat jejunal crypt cells are contradictory. According to Tutton and Helme (1974) a single injection of propranolol or practolol (10 mg/kg) increased the mitotic index twofold and shortened the duration of the cell cycle of the crypt cells. However, upon repeating the experiments with double the dose of propranolol, Maurer-Schultze et al. (1986) observed no such effects using cell kinetic methods with 3H-thymidine instead of the stathmokinetic method applied by Tutton and Helme. Since the discrepancy in the results may have been due to methodological differences the same stathmokinetic method used by Tutton and Helme has been applied in the present work. However, the results obtained with this method indicate no influence by propranolol on the proliferation of jejunal crypt cells even with a dose of 20 mg/kg. Consequently we were unable to confirm the stimulant effect of propranolol on crypt cell proliferation. The possible causes of the discrepancy between the present results and those of Tutton and Helme are discussed.

Animals↗
PubMed1987

Effects of chronic administration of a monoclonal antibody against human renin in the marmoset.

In this study, the hypotensive efficacy of R-3-36-16, a monoclonal antibody against human kidney renin, was investigated during chronic administration to a primate. R-3-36-16 was given by continuous intraperitoneal infusion with osmotic minipumps to normotensive marmosets fed a low-sodium diet in doses of 30 or 300 micrograms/kg/day for 14 days. The lower dose had no effect on blood pressure (BP) or plasma renin activity (PRA). After two days of treatment, the higher dose reduced PRA by 57% and lowered BP by 13 +/- 7 mm Hg. Although the hypotensive response persisted after 14 days of treatment (-17 +/- 2 mm Hg), PRA had recovered to pretreatment levels. BP gradually returned to pretreatment values in the week after stopping the treatment. There was no evidence of an immune reaction when an acute challenge dose of R-3-36-16 was given 7 weeks after stopping the chronic treatment. Thus, R-3-36-16 appears to be an effective and well-tolerated hypotensive agent during chronic administration to sodium-depleted primates. The hypotensive response does not seem to be directly related to the inhibition of renin in the plasma.

Animals↗
PubMed1986

Influence of the beta-adrenergic antagonists propranolol, practolol and oxprenolol on the proliferation of rat jejunal crypt cells.

Beta-adrenergic blockade by quite large doses of propranolol, practolol and oxprenolol, once or continuously applied, does not influence jejunal crypt-cell proliferation in the rat. After a single i.p. injection of 20 mg/kg propranolol or practolol and even of 100 mg/kg practolol, the mitotic index, the labelling index and the duration of the S phase do not differ between treated and untreated control animals nor between animals treated with the different drugs. Continuous application of 30 mg/kg/d propranolol, practolol or oxprenolol for 7 or 14 days does not affect the mitotic and labelling indices either, nor does it change the duration of the cycle of the jejunal crypt cells and its phases as determined by the percent labelled mitoses method. These results are in contrast to those reported previously by Tutton & Helme (1974).

Adrenergic beta-Antagonists↗
PubMed1979

Localization of lactate dehydrogenase isozymes in human muscle tissues by the mixed aggregation immunocytochemical technique.

Lactate dehydrogenase (LDH) isozyme composition and localization was determined in sections of skeletal, heart and smooth muscle by the mixed aggregation immunocytochemical method using first antibody directed against purified human LDH-A4 (M4) or LDH-B4 (H4) followed by the enzymes LDH-A4 and LDH-B4, respectively. An even distribution of the two monomers in all fibres was seen with heart muscle and smooth muscle. Heart muscle had a low concentration of A-monomers and a high concentration of B-monomers, whereas the smooth muscle had equal concentrations of the two monomers. In contrast, skeletal muscle from m. quadriceps femoris was found to be composed of two muscle fibre types, one containing mainly A-, the other mainly B-monomers. On the basis of succinate dehydrogenase activity it was shown that the red (type 1) fibres contain mainly B-monomers and the white (type 2) fibres mainly A-monomers of LDH.

Female↗
PubMed1979

Assessment of immunocytochemical techniques with particular reference to the mixed-aggregation immunocytochemical technique.

Antibody-mediated assays are believed to be the most sensitive and specific techniques for antigen localization available to date, but they need extensive controls if specific and quantitative results are to be obtained. Problems arise from both the antibody source and the antigen in the tissue preparation. Specificity of the antibody is often difficult to prove in any direct or indirect antibody technique. When markers are coupled to antibody, binding and diffusion properties of the antibody are changed. Diffusion of antigen from its site and loss of antigenic determinants due to procedures for preserving morphology are two other main problems. The mixed-aggregation immunocytochemical technique provides another way of checking the selectivity of the antibody, since a cross-reactive antigen introduced into the section and not the antibody itself is what is seen. Quantitative studies using localization of lactic dehydrogenase isoenzymes in human skeletal muscle fibres show that the technique fulfils most of the present criteria on precision, specificity and validity for determination of enzymes. It also ensures that more than 80% native antigen in frozen sections is fixed and is two to three times more sensitive and selective than the direct antibody technique.

Antibody Specificity↗
PubMed1979

Relevance of enzyme evaluations in 24h urine to rat kidney injury caused by i.v. cephaloridine injection.

Male rats were housed singly in metabolic cages, injected i.v. with cephaloridine, 24 h urine samples collected successively; then the rats were killed for obtaining the kidneys of corresponding animals. The concentrations of protein, aminopeptidase (AP), alkaline phosphatase (aPP), lactic dehydrogenase (LDH), and aldolase (ALD) were determined in urine and the percentages of injured proximal tubules counted in sections stained for aPP. The results from individual animals were: (1) After placing animals singly in metabolic cages large but not systematic changes of urinary enzyme concentrations occurred. After 6-10 days the enzymes reached steady state levels. (2) After a single injection of cephaloridine a dose dependent injury of proximal tubules was observed, the urinary LDH content correlating best with the tubular injury (r greater than 0.93) and giving up to 1,000 fold increases above normal values. (3) A circadian rhythm of the susceptibility of rat kidney for cephaloridine was observed, the smallest response was seen when the animals were injected at 7 a.m. and the largest after injection at 7 p.m. (4) In subacute toxicity studies urinary LDH was increased on day 2 above the extent after a single dose, but declined on day 3 to reach normal levels after 8 to 10 days (time of sacrifice). The kidneys revealed practically normal histology. The other enzymes studied had also returned to normal values. This indicates some adaptation mechanism.

Alkaline Phosphatase↗