Combined Aldehyde-Fuchsin and Periodic Acid-Schiff Staining of the Pituitary

Aldehyde-fuchsin and the periodic acid-Schiff procedure can be applied in sequence to the same tissue section and combined with a stain for acidophils, such as orange G, for routine analysis of the pituitary. This is made possible by fixation in a mixture containing 5% chrome-alum, 5% HgCl₂ and 5% formalin; control of the pH and the activity of the aldehyde-fuchsin; and increased sensitivity of the Schiff reagent, effected by reducing its SO₂ content. Although designed especially for the pituitary, the procedure is also applicable to other histological problems.     

Clostridium perfringens in Meat and Meat Products

A total of 262 specimens of meat and meat dishes were examined for the presence of Clostridium perfringens. Of this total, 161 were raw, unprocessed beef, veal, lamb, pork, or chicken; 101 were processed meats and meat dishes. C. perfringens was isolated from 113 (43.1%) of these specimens. The highest percentage of contamination (82%) was found in veal cuts, and the lowest (4.7%) in sliced sandwich meats and spreads. Only 2 of the 113 isolates were shown to produce heat-resistant spores, which indicates a very low incidence (0.8%) of contamination. These findings indicate that outbreaks of C. perfringens food-borne disease in the Cincinnati area are caused principally by the contamination of the food with vegetative cells or spores of the organism after cooking. Studies of the effects of various holding temperatures on the growth of C. perfringens indicated that, in the range of 5 to 15 C, no multiplication would occur, but that viable cells would still be present at the end of a 5-day holding period. Extremely rapid growth occurred at temperatures around 45 C, and complete inhibition of growth was accomplished between 49 and 52 C.     

Das piezoelektrische Verhalten der menschlichen Zahnhartgewebe

Zusammenfassung Bei menschlichen Zähnen wurde piezoelektrisches Verhalten festgestellt; dieses ist gebunden an den Kollagenfaseranteil des Dentins und des Zahnzementes. Schmelz ist nicht piezoelektrisch; entmineralisierte Zähne zeigen ein verstärktes piezoelektrisches Verhalten.Es besteht eine piezoelektrische Achse in der physiologischen Zahnlängsrichtung, deren Richtungssinn bei allen Zähnen des Ober- und Unterkiefers gleich ist und offenbar der morphogenetischen Entwicklungsrichtung des Zahnes entspricht. Außerdem besteht rund um die piezoelektrische Längsachse herum in allen radialen Richtungen zwischen Pulpahöhle und Zahnaußenseite piezoelektrisches Verhalten, das möglicherweise in Beziehung zur Struktur der Dentinlamellen und damit zur morphogenetischen Zahnentwicklung in radialer Richtung steht.Die piezoelektrischen Effekte zeigen von Zahn zu Zahn eine erhebliche biologische Streubreite. Bei unserem Untersuchungsmaterial lagen die Effekte zwischen 3,5·10^–10 und 1,9·10^–9 est E Ldg./dyn. Diese Werte entsprechen etwa 0,5 bis 2,8% des piezoelektrischen Koeffizienten d₁₁ von -Quarz.

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Summary Human teeth are piezoelectric. This is due to the collagen-fibril content of the dentine and tooth cement. Enamel is not piezoelectric; decalcified teeth show increased piezoelectric behaviour.There is a piezoelectrical axis corresponding to the physiological longitudinal tooth axis, which is in the same direction in all teeth of both upper and lower jaws, and which seems to correspond to the morphogenetical development axis of teeth. Piezoelectric behaviour also exists between the pulp cavity and outer surface of the tooth in all directions radial to the piezoelectric longitudinal axis. The latter is possibly due to the dentine lamellae, and therefore may be connected with the radial direction of morphogenetical tooth development.The piezoelectric effects show large biological variations in different teeth. The effects from our research materials vary between 3.5·10^–10 and 1.9·10^–9 e.s.u./dynes. These results correspond to about 0.5 to 2.8% of the piezoelectrical coefficient d₁₁ of -quartz.

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Unterstützt durch Sachbeihilfen der Deutschen Forschungsgemeinschaft.

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cand. med. dent. und derzeit Doktorand an der Klinik und Poliklinik für Zahn-, Mund- und Kieferkrankheiten, Universität Kiel.     

Localization of melanin synthesis within the pigment cell: Determination by a combination of electron microscopic autoradiography and topographic planimetry

Summary Localization of melanin synthesis within the pigment cells of the Cloudman S-91 mouse melanoma was determined by means of a combination of high resolution autoradiography and topographic planimetry. Initial melanin biosynthesis occurred predominantly in the endoplasmic reticulum and associated ribonucleoprotein particles of the melanocytes. By measuring a number of cell organelles and employing the index of relative specific localization it could be shown that the nucleus and mitochondria are of little or no importance in the process of melanogenesis.This investigation has been supported by the following research grants: CA 06548 CB, NIH, PHS and an Institutional Grant from the American Cancer Society (to H. M. H.); CA-05887, NIH, PHS (to A. S. Z.); M-00388 and NB-00782, NIH, PHS (to J. F. H.). One of us (H. M. H.) holds a Research Cancer Development Award (5-K 3-GM-2634) of the National Institute of General Medical Sciences, Public Health Service.We are grateful to Mr. Ronald Abler for his help with the topographic measurements; to Dr. Erhard Haus for help and advice; to Mr. J. Thornby and Mr. A. P. Basu for assistance with the statistical aspects of this study; and to Mrs. Lenore Mottaz, Miss Bernice Uittenbogaard, and Mrs. Judith Strong for careful technical assistance.     

Semisynthesis of Arg15, Glu15, Met15, and Nle15-aprotinin involving enzymatic peptide bond resynthesis

The semisynthesis of homologues of aprotinin, the bovine pancreatic trypsin inhibitor, is described. The P₁ lysine¹⁵ residue was replaced by two methods. The first procedure, which consisted of two enzymatic steps for the incorporation of other amino acids has previously been described. The second approach consisted of six steps of both enzymatic and chemical nature. The modified inhibitor, in which the lysine¹⁵-alanine¹⁶ peptide bond is hydrolyzed, was used as the starting material. All carboxyl groups of the modified inhibitor were esterified with methanol; the lysine¹⁵ methylester group was then selectively hydrolyzed. Afterward, lysine¹⁵ itself was split off. Arginine, glutamic acid, methionine, andl-2-aminohexanoic acid (norleucine, Nle) were incorporated using water-soluble carbodiimide combined with an acylation catalyst. The methylester group was used to prevent polymerization. The reactive-site peptide bonds were resynthesized using either chymotrypsin or trypsin.     

Photoperiodic influences on sexual behavior in male Syrian hamsters

The effect of photoperiodic conditions on sexual behavior was investigated in male Syrian hamsters that were either gonadally intact, or castrated and treated with low doses of testosterone throughout the experiment. Hamsters were exposed to long (LD 16:8) or short (LD 8:16) days for 7 weeks; for the next 8 weeks, either they were exposed to an intermediate daylength (LD 12:12), or daylength conditions remained unchanged. Sexual behavior was affected by photoperiod conditions in both gonadally intact animals and testosterone-treated castrates, but to different degrees. Intact males exposed to short days for 15 weeks exhibited gonadal regression, and their copulatory performance was impaired. The percentage of animals that intromitted or ejaculated was significantly reduced. Additional measures of sexual performance among the copulating males were also affected. In contrast, among the castrates with testosterone clamped at low but stable levels, the proportion of males that mounted, intromitted, or ejaculated was not affected by photoperiod. However, among the males that continued to copulate, sexual performance changes were present in the short-day castrates that resembled those displayed by the intact males. We infer that these behavioral effects in both hormonal conditions reflect primarily a difficulty in the attainment of intromission. Gonadal regression alone cannot easily account for the behavioral deficits of the intact males, because circulating testosterone levels at the end of the experiment were not significantly different between the gonadally intact hamsters and the castrated, testosterone-treated hamsters exposed continuously to short days. Males transferred from either long or short days to the intermediate-daylength condition responded behaviorally to this photoperiod as if it were a short day, that is, their ejaculatory frequency declined. We conclude that male hamsters exposed to photoinhibitory daylengths exhibit deficits in their sexual behavior, not only because endogenous levels of testosterone decrease, but also because the substrates on which this hormone acts become less responsive. We hypothesize that under physiological conditions, the episodic secretion of testosterone imposes constraints on the maintenance or restoration of copulation, and that the potent behavioral effects achieved by constant-release implants of testosterone may mask the presence of photoperiodically induced alterations in the hamster's sensitivity to this gonadal hormone.     

1-Methyl-4-Phenyl-1,2,3,6-Tetrahydropyridine- and 1-Methyl-4-(2''-Ethylphenyl)-1,2,3,6-Tetrahydropyridine-Induced Toxicity in PC 12 Cells: Role of Monoamine Oxidase A

The toxicity of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), 1-methyl-4-(2'-ethylphenyl)-1,2,3,6-tetrahydropyridine (2'Et-MPTP), and their corresponding pyridinium species was studied in the rat pheochromocytoma PC12 cell line. MPTP and its analogues are known to be metabolized by monoamine oxidase (MAO) to dihydropyridinium intermediates which are further transformed, either enzymatically or spontaneously, into pyridinium species. MAO activity in PC12 cells is almost exclusively of the A form, and 2'Et-MPTP is a good substrate for both MAO-A and MAO-B. In contrast, MPTP is a poor substrate for MAO-A, but a good substrate for MAO-B. 2'Et-MPTP caused considerably more cell death than MPTP in the PC12 cells. However, 1-methyl-4-(2'-ethylphenyl)pyridinium and 1-methyl-4-phenylpyridinium, the corresponding pyridinium species formed from 2'Et-MPTP and MPTP, respectively, were equipotent as toxins. The toxic effects of the tetrahydropyridines and their corresponding pyridiniums were both concentration- and time-dependent. Measurements of the levels of the pyridinium species formed and the remaining tetrahydropyridine in the media indicated that 2'Et-MPTP was converted about five to seven times more readily into its toxic pyridinium species than was MPTP. There was, moreover, an excellent correlation between amount of pyridinium formed and cell death. There was also a parallel between the capacity of clorgyline and pargyline, irreversible MAO inhibitors, to decrease the formation of the pyridinium species and their capacity to protect against the toxic actions of the tetrahydropyridines. These data are consistent with the concept that the MAO-A-dependent formation of the pyridinium species from the tetrahydropyridine is a prerequisite for toxicity in PC12 cells.