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Previous attempts to prepare skeletal muscle basal laminae (BL) for ultrastructural analyses have been hampered by difficulties in successfully removing skeletal muscle proteins and cellular debris from BL tubes. In the present study we describe a two phase method which results in an acellular muscle preparation, the BL of which are examined by light, transmission electron, and scanning electron microscopy. In the first phase, excised rat extensor digitorum longus muscles are subjected to x-radiation and then soaked in Marcaine to inhibit muscle regeneration and to destroy peripheral muscle fibers. The muscles are then grafted back into their original sites and allowed to remain in place 7-14 days to allow for maximal removal of degenerating muscle tissue with minimal scar tissue formation. In the second phase, the muscle grafts are subjected sequentially to EDTA, triton X-100, DNAase, and sodium deoxycholate to remove phagocytizing cells and associated degenerating muscle tissue. These procedures result in translucent, acellular muscle grafts which show numerous empty tubes of BL backed by endomysial collagenous fibers. These preparations should be useful for morphological analyses of isolated muscle BL and for possible in vitro studies by which the biological activity of muscle BL can be examined.
Anat Rec 1991 Jul
PMID:A method for preparing skeletal muscle fiber basal laminae. 190 15

An interrelationship between atrial natriuretic peptide (ANP) and the renin-angiotensin system has been established. Both of these hormonal systems are modulated by sodium balance. The role of the beta-adrenoceptor in the regulation of release of ANP is not clear. We therefore undertook a study to examine changes in atrial-specific granule number and plasma ANP level following beta-adrenoceptor blockade in rats on low and high sodium intakes. A low-sodium diet, as compared with a high-sodium diet, elevated right and left atrial-specific granule number (right atria 54.6 +/- 8.7 vs. 42.3 +/- 5.7; left atria 47.7 +/- 7.7 vs. 30.6 +/- 3.4 granules/unit area) and plasma renin activity (28 +/- 3.7 vs. 5.4 +/- 0.8 ng AI/ml/hr). Plasma ANP levels were lower in the low-sodium animals (98 +/- 34 vs. 345 +/- 38 pg/ml). When treated with the nonspecific beta-adrenoceptor blocker propranolol, the elevated plasma renin activity and atrial-specific granule number in rats on a low sodium intake were significantly less. Neither of these parameters changed in rats on a high sodium intake. Conversely, propranolol treatment resulted in lower plasma ANP levels in rats with high sodium intake. The already-suppressed plasma ANP level in rats on a low-sodium diet was unaltered with beta-adrenoceptor blockade. The results suggest that dietary sodium intake is an important determinant of the response of atrial-specific granule number and plasma ANP levels following beta-adrenoceptor blockade with propranolol.
Anat Rec 1990 Dec
PMID:Atrial-specific granule number and plasma atrial natriuretic peptide in rats: effects of beta-adrenoceptor blockade and sodium intake. 198 Sep 94

Eight endodontic agents were tested for their reactivity with DNA by the rec-assay system. Employing 2 strains of Bacillus subtilis with different recombination capacities, the assay suggested that DNA damage is produced by agents for root canal cleaning containing sodium hypochloride. Other endodontic drugs, including disinfectants for caries cavities, sedatives for pulp, root canal disinfectants, and pulp devitalizing agents containing phenol, camphor, tricresol, formalin, and paraformaldehyde were also positive by rec-assay and would seem to potentially of damage cellular DNA in Bacillus subtilis. Though no clear evidence exists that drugs showing positive results by rec-assay induce malignant tumors in the oral region, these agents should be used carefully since they would seem to possibly cause genetic toxicity in mammalian cells.
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PMID:Investigation of DNA reactivity of endodontic agents by rec-assay. 213 71

Our recent observation that the basement membranes of brain microvessels do not stain with the cationic dye ruthenium red has raised the question of whether the basement membranes of this and other vascular beds functioning as barriers between blood and neural tissues are deficient in the polyanionic macromolecules, such as glycosaminoglycans, which are responsible for the ruthenium red staining of other vascular basement membranes. We therefore attempted to produce staining in the only barrier-type microvascular basement membrane known to contain heparan sulfate. Bovine retinas were fixed by immersion in aldehyde fixatives containing ruthenium red, buffered with either 10 mM or 100 mM sodium cacodylate. We found discrete, electron-dense deposits of ruthenium red in vascular basement membranes, quite similar to those seen in vascular basement membranes of nonneural tissues after exposure to ruthenium red. These deposits were more distinct and more frequent in tissue exposed to ruthenium red-aldehyde solutions buffered with 10 mM cacodylate. They were not seen if ruthenium red was omitted from the fixative. The results demonstrate that anionic macromolecules in basement membranes of barrier-type microvessels can be stained with cationic dyes, and suggest that the failure of brain microvessels to stain with ruthenium red may be the result of a relative or total lack of polyanion in this basement membrane, or of other unique properties.
Anat Rec 1987 Dec
PMID:Ultrastructural studies of bovine retinal microvascular basement membranes with the cationic dye ruthenium red. 245 89

Bovine spongiform encephalopathy probably results from the use of commercial diets containing scrapie-contaminated ingredients. Of all the chemical and physical decontamination procedures which are effective against conventional viruses, only high temperature autoclaving, high concentrations of sodium hypochlorite, and possibly molar sodium hydroxide, are useful against the group of unconventional transmissible agents which includes scrapie. The implications of this problem for the rendering industry and farm practice are discussed.
Vet Rec 1989 Mar 25
PMID:Scrapie agent decontamination: implications for bovine spongiform encephalopathy. 249 90

One hundred and eleven heifers and cows with caecal dilatation and torsion were examined and their subsequent progress monitored. Using various criteria it was decided whether the animal was to be slaughtered or treated conservatively or surgically. Conservative treatment consisted of a continuous drip infusion containing neostigmine and of purgatives such as liquid paraffin, sometimes in combination with sodium sulphate. Surgical treatment consisted of laparotomy on the right flank with emptying and sometimes partial resection of the caecum. Five animals had to be slaughtered before or during surgery and another five which developed severe peritonitis were slaughtered after surgery. Fifty-nine animals underwent surgery once without resection of the caecum and 20 with. Another 14 animals needed surgery twice.
Vet Rec 1989 Oct 21
PMID:Therapy and clinical progress of cattle with dilatation and torsion of the caecum. 259 75

A microbiological procedure for determining dioxidine concentrations in biological fluids with using E. coli AB 2472 rec A 16, a reparation deficient strain as a test organism is described. Cell suspension of the strain 24-hour culture is added to 1.2 per cent agar with Hottinger digest (140 mg per cent of amine nitrogen), 3 g/l of disubstituted sodium phosphate and 0.4 per cent of glucose cooled to 50 degrees C. 10 ml of the medium are added to every Petri dish with metallic cylinders put on the agar. After the medium solidification the cylinders are removed and 0.1 ml of the solution being tested is added to every well. The dishes are incubated for 24 hours under anaerobic conditions. The test system sensitivity is 0.2 microgram/ml of dioxidine. The relationship between the growth inhibition zone and the drug concentration is linear within dioxidine concentrations of 0.2 to 3.2 micrograms/ml.
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PMID:[A microbiological test system for determining dioxidine levels in biological fluids]. 266 75

A weaner ration containing carbadox at concentrations of 331 to 363 mg/kg was accidentally fed to suckling and weaned pigs in an 84 sow herd. Discarded ration was fed to 36 sows. One hundred and sixty five weaner pigs died in a 10 week period with clinical signs including refusal to eat, ill thrift, the passing of hard pelleted faeces, posterior paresis and death in seven to nine days. The surviving weaners did not thrive and some males showed poor testicular development. Sows and suckling pigs that consumed the ration also failed to thrive as did the progeny of affected sows. The main pathological finding was obliteration of the zona glomerulosa of the adrenal cortex. Increased potassium and decreased sodium concentrations in serum were the most notable and consistent biochemical findings.
Vet Rec 1989 Apr 08
PMID:Accidental carbadox overdosage in pigs in an Irish weaner-producing herd. 271 36

When they were turned out to grass in May 1987 for their first season, 10 calves were dosed with a 5 x 750 mg oxfendazole pulse release bolus (OPRB) and a monensin sodium rumen delivery device (RDD); eight calves received one OPRB; 10 calves received one RDD and eight calves received neither bolus. Each group was set-stocked on individual paddocks which had been grazed during the previous season by cattle which developed clinical parasitic gastroenteritis and bronchitis (husk). In July, before they were due to be moved to new pastures in mid-summer, and before they were dosed strategically with levamisole HCl, some of the calves not dosed with an OPRB succumbed to clinical parasitic gastroenteritis and husk and received emergency anthelmintic treatment, after which no further clinical episodes occurred. The 'dose and move' strategy was implemented in early August after which both groups not dosed with an OPRB were set-stocked together until the trial ended on October 14, 147 days after turn out. The two groups of calves which had received the OPRB were also moved to new pasture and set-stocked together until the end of the trial. No evidence of clinical helminthiases developed in either of the two groups of calves dosed with OPRBs and their faecal worm egg and larval counts, and plasma pepsinogen activities remained low. They gained significantly more weight than the two groups of calves not dosed with OPRBs (P less than 0.001). The bolus types were compatible and induced no untoward side-effects when used together.
Vet Rec 1989 Jul 15
PMID:Concurrent use of the oxfendazole pulse release bolus and the monensin rumen delivery device in young grazing cattle. 277 30

The pattern and timing of the breakdown and loss of matrix proteins were studied in developing rat incisor enamel using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), fluorography, radioautography, and in vitro incubations of proteins isolated from freshly dissected, crushed pieces of enamel. For biochemical studies, the technique of Robinson et al. (1974, 1977, 1983) was used to transect the enamel organ and enamel into a series of strips at 1 mm intervals along the length of the tooth. The proteins in each strip were extracted and either quantified by Lowry analysis or applied to 12% slab (enamel) or 5-15% continuous gradient (enamel organ) SDS-polyacrylamide gels and separated by electrophoresis. The biochemical studies indicated that the amount of protein contained within an enamel strip increased gradually by volume across the secretory stage, reached a peak early during the maturation stage, and then declined rapidly thereafter. The distribution of enamel proteins on SDS-polyacrylamide gels changed markedly throughout this period. These changes included increases and decreases in the intensity of staining of proteins at certain molecular weights (e.g., 18 kDa) and the appearance and disappearance of some proteins not seen clearly near the start of the secretory stage of amelogenesis (e.g., 32 and 10 kDa). Labeling studies with 35S-methionine suggested that the "stacked" arrangement of proteins typical of forming enamel (secretory stage) actually represented a very dynamic association of proteins, with new ones being added at the top of the stack and then breaking down with time to become those seen at lower molecular weights. Across the secretory stage, new proteins were always added to the top of the stack, but during early maturation this activity slowed dramatically, allowing the breakdown of aging proteins to be visualized more clearly. Radioautographic studies with 3H-methionine indicated that the breakdown of newly secreted proteins also was correlated with a movement of label from the site of secretion into deeper, previously unlabeled, areas of forming enamel. In vitro studies revealed that the rate and degree of breakdown of enamel proteins varied markedly, depending on the stage of amelogenesis from which the proteins were extracted. Secretory stage enamel proteins showed slow in vitro degradation with accumulation of proteins near 18 kDa. Early maturation stage enamel proteins showed more rapid breakdown with little accumulation of proteins near 18 kDa, whereas late maturation stage enamel proteins showed complete degradation by 2 days of incubation in vitro.(ABSTRACT TRUNCATED AT 400 WORDS)
Anat Rec 1989 Jun
PMID:Degradation and loss of matrix proteins from developing enamel. 277 8


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