Histochemical detection of l-gulonolactone: phenazine methosulfate oxidoreductase activity in several mammals with special reference to synthesis of vitamin C in primates

Summary An improved detection of activity of l-gulonolactone oxidase, which is responsible for the final oxidative step in the synthetic process of l-ascorbate from glucose in animals, was achieved using phenazine methosulfate and cyanide. Cold acetone fixation eliminated non-specific deposition of formazan on lipid droplets. The specificity of the method was tested and proven by a biological control, histochemical controls, inhibitors and activators. By application of the method, strong reactivity was found in the cytoplasm of centrilobular parenchymal cells of livers of the opossum, rat, ground squirrel and flying squirrel. Staining of dog liver was moderate and centrilobular. Prosimians were strongly positive: The centrilobular localization was found in the tree shrew and galago; slow lorises and some pottos showed strong reactivity in centrilobular cells and some peripheral cells as well. These prosimians seem to be able to synthesize l-ascorbate as many lower mammals are. On the contrary, true simians (i.e. the squirrel monkey, spider monkey, rhesus monkey and chimpanzee) were negative as guinea pigs were, suggesting their probable inability for l-ascorbate synthesis.Visiting scientist from the Department of Anatomy, Tokyo Medical and Dental University, Tokyo, Japan. T. R. Shanthaveerappa in previous publications, also fellow, Department of Anesthesiology, Emory University.     

Histochemical studies on urate oxidase in several mammals with special reference to uricolytic ability of primates

Summary Strong reactivity for urate oxidase was found in the liver parenchymal cells of the prosimians (i.e. the tree shrew, slow loris, potto and galago) as well as those of lower mammals. The liver parenchymal cells of the platyrrhine monkeys (i.e. the marmoset, owl monkey, squirrel monkey, capuchin monkey and spider monkey) were moderately positive. There was no preferential distribution of granular reaction products in zones of liver lobules of these species. The prosimians and platyrrhine monkeys seem to be uricolytic as lower mammals are. On the other hand, the old world monkeys (i.e. Java monkey and rhesus monkey) and the apes (i.e. the orang-utan and chimpanzee) were histochemically negative.     

Development of the bitterling,Paracheilognathus himantegus (Cyprinidae), with a note on minute tubercles on the skin surface

The development of the eggs and larvae and minute tubercles on the skin surface ofParacheilognathus himantegus larvae were observed. The egg began to hatch approximately 68 hours after insemination and the larvae reached the free-swimming stage 23 days after hatching at water temperature of 22±1°C. The larval development and minute tubercles on the skin surface of this species were similar to those ofAcheilognathus lanceolata, A. limbata, A. signifer andTanakia tanago. However, the shape of the ripe eggs ofP. himantegus differed from those of the four species. As regards the shape of eggs, there was a common characteristic amongP. himantegus, Rhodeus uyekii andA. limbata from Korea. As regards larval development,P. himantegus had two characters also found inRhodeus. These facts seem to suggest thatP. himantegus is closely related toA. lanceolata, A. limbata, A. signifer andT. tanago but is more specialized than these four species, except forA. limbata from Korea.     

Histochemical demonstration of O-glycosidically linked,type 3 based ABH antigens in human pancreas using lectin staining and glycosidase digestion procedures

Summary Histochemical analyses of the chemical structures of sugar sequences with or without blood group specificity were carried out by combined stepwise digestion of tissue sections with exo-and endoglycosidases and subsequent lectin stainings in formalin-fixed, paraffin-embedded human pancreas. In acinar cells from blood group A or AB secretor individuals, sequential digestion with -N-acetylgalactosaminidase and -L-fucosidase imparted reactivity with peanut agglutinin (PNA) in cells reactive with Dolichos biflorus agglutinin as well as those with Ulex europaeus agglutinin I(UEA-I). Simple fucosidase digestion imparted the PNA reactivity only in UEA-I reactive cells. Sequential digestion with -galactosidase and fucosidase likewise liberated the PNA binding sites in Griffonia simplicifolia agglutinin I-B₄ reactive cells from blood group B and AB secretors. Sialidase digestion liberated the PNA binding sites not only in acinar cells but also intercalated duct cells, islet cells of Langerhans and endothelial cells. The PNA reactivity obtained by these enzyme digestions was eliminted by endo--N-acetylgalactosaminidase (endo-GalNAcdase) digestion. Preexisting PNA affinity in acinar cells from nonsecretors was also susceptible to endo-GalNAcdase treatment. Following the endo-GalNAcdase digestion, fucosidase or sialidase digestion recovered the PNA reactivity in acinar cells from nonsecretors. These results show that ABH determinants carried on O-glycosidically linked type 3 chain (D-galactose-(1-3)-N-acetyl-D-galactosamine1-serine or threonine) are secreted in pancreatic acinar cells and suggest that product coded by the secretor gene is required for the complete conversion of type 3 precursor chains into H determinants.     

Purification and spectral characterization of a 121-kilodalton phytochrome from etiolated pea (Pisum sativum L.) seedlings

Procedures for the purification of native phytochrome from etiolatedpea seedlings without the use of immuno-purification techniquesare described. Phytochrome (in the P_FR form) was purified bypolyethyleneglycol fractionation, adsorption to pentyl agaroseand batch elution, chromatography on DEAE-Sepharose, adsorptionto phenyl Toyopearl and batch elution, and chromatography onRed Toyopearl. The resulting phytochrome had specific absorbanceratios (SAR = A₆₆₆/A₂₈₀ of P_R) that ranged from 0.55 to 0.6.The subsequent chromatography on Sephacryl S-300 yielded verypure phytochrome with a SAR of 0.98. P_R and P_FR peaks in thedifference spectrum of the phytochrome were centered at 665and 730 nm, respectively. The spectral change ratio (Ar/Afr)of the difference spectrum was unchanged after the chromatographyon phenyl Toyopearl, and the value was 1.05–1.08, indicatingthat the spectral properties of this preparation were intact.The absorption spectra indicated that the peak absorbance ofP_FR was at 728–730 nm and that of P_R was at 666–667nm. These peak positions were essentially same as those obtainedwith the undegraded oat phytochrome. Incubation of the samplepurified on Sephacryl S-300 at 25?C for 5 h in either the P_Ror P_FR form did not result in degradation of the molecule. Therate of dark reversion of P_FR observed with the purified peaphytochrome was similar to that observed in vivo. The additionof dithionite had no effect on the reversion rate. ²Present address: Fuji-Gotenba, Research Lab. of Chugai PharmaceuticalCo. Ltd., Gotenba, Shizuoka, 412 Japan (Received February 22, 1990; Accepted May 28, 1990)     

Tracheoplasty with palatal mucoperiosteal graft

A case of tracheal reconstruction with a palatal mucoperiosteal graft is reported. The patient is a 75-year-old woman who had the anterior wall of her trachea resected for invading thyroid carcinoma. Palatal mucoperiosteum, which is easy to handle, provided both the lining and supporting tissue for the trachea. Local neck skin was used for covering. This procedure proved very successful.     

Augmentation of the nostril splint for retaining the corrected contour of the cleft lip nose

Whatever method is used to correct the deformity of the cleft lip nose, it is important to maintain the corrected contour of the nose for a certain period postoperatively. For this purpose, moldable silicone rubber is used to add volume to a ready-made nostril splint. With this material it is easy to make a splint that fits the individual contour of the nostril. Clinical examples are presented.