Basic and applied histochemistry最新文献

Histochemical techniques were applied to salivary glands removed from adult multimmate rodents (Praomys) of either sex to detect and localize the following enzymatic activities: acid and alkaline phosphatase, arylsulphatase, ali-esterases, beta-glucuronidase, N-acetyl-betaglucosaminidase, and L-leucyl-aminopeptidase. No reaction was observed for alkaline phosphatase and glucuronidase. The glands reacted differently to the other enzymatic activities. Alkaline phosphatase and glucosaminidase were present only in one glandular type whereas arysulphatase and esterases were present in all types although demonstrating a variable staining intensity in different glands. Sharp differences in some enzymatic activities of the submandibular and parotid glands were related to the sex of the animal.

The localization of substance P is studied in the human prevertebral sympathetic ganglia and descending colon by the indirect immunofluorescence technique. Strongly immunoreactive axons reach the coeliac, superior mesenteric and inferior mesenteric ganglia. The sympathetic postganglionic neurones are surrounded by a meshwork of positive beaded fibers and terminals. Varicose nerve fibers are variously distributed in all nonepithelial layers of the colon. A close network of fibers and terminal surrounds the ganglionic cells in the mienteric and submucous plexuses. Substance P-like immunoreactive perikarya are present in these structures.

Rat brain thiamine pyrophosphatase (TPPase) was separated by thin layer polyacrylamide gel isoelectric focusing (IF) and stained with a modification of the lead conversion method of Allen and Hyncik (1963). 10 bands with TPPase activity were observed in the pH range 4.6-7.1. The overall IF pattern of TPPase was similar to that of uridine diphosphatase and inosine diphosphatase but was clearly different from that of adenosine diphosphatase, p-nitrophenyl phosphatase, alpha-naphthyl phosphatase and thiamine monophosphatase. A semiquantitative assessment of TPPase isoenzymes has been performed using laser densitometry.

With the help of a computer program using Newton's iterative method, it is possible to obtain a practically instantaneous solution for the two-wavelenght method equation even for values different from 2 for the ratio k of the extinction coefficients. The influence of the extinction ratio and of the mean transmittance on the correction of the distributional error and on the propagation of stochastic and systematic errors into the computed chromophore amount has been studied. After due precautions are taken to minimize chromatic and non-specific light loss errors, the selection of the optimal wavelenght couple will basically depend only on the compromise between correction of the distributional error and the least propagation of stochastic errors into the computed chromophore amount. In many instances, the use of interference filters will be convenient, since their use will eliminate the uncertainties of monochromator readjustment and will allow evenly illuminated field to be obtained more easily, particularly in the case of large measuring fields.

Immunohistochemical expression of 8 cases of mucoepidermoid carcinomas (G-I, 3 cases; G-II, 2 cases; and G-III, 3 cases) revealed marked heterogeneity of the proteins examined. Immunohistochemically detectable keratins (TK, KL1, and PKK1) were distributed in epidermoid cells, but were absent in mucous secreting cells. Strongly positive deposits of keratin proteins were detected in squamoid tumor cells in the G-I tumors. The tumor cells displayed positive staining for S-100 alpha, but did not stain with polyclonal S-100 antiserum or with monoclonal S-100 beta. The cells showing highest reactivity for S-100 protein were scattered in neoplastic foci and were probably Langerhans cells. Lactoferrin and lysozyme reactions were generally negative in tumor foci; but a positive reaction for lactoferrin was found in luminal tumor cells although rarely, and lysozyme staining was occasionally noted in histiocytes in the stroma. Amylase activity was usually absent in the tumor cells, with the exception of one case in which it was confined to the tumor cells. Mucoepidermoid carcinomas of various grades indicated marked heterogeneity in terms of various immunohistochemically detectable proteins.

This immunocytochemical study of the late postnatal development of the medio-basal hypothalamus revealed the presence of ACTH 1-39 like positivity in neurons of the arcuate nucleus form the begin of this study (day E 18-20) onwards. Alpha MSH positivity, on the contrary, is not present in cells of the same area before day P 16. No other areas in the developing medio-basal hypothalamus contain perikaryal positivity for alpha M-SH or ACTH 1-39. The pituitary contains ACTH 1-39 like positivity from the begin of this study (day E 18-20) onwards. Fibers are positive for alpha MSH during the fetal development of the medio-basal hypothalamus, demonstrating an overal reactivity without varicosities and restricted to bundles or neuropil areas. Towards P 16 the alpha MSH positivity diminishes in the whole medio-basal hypothalamus, remaining present only in large fibre systems like the fornix. ACTH 1-39 like fiber positivity is already distributed in arcuate and periventricular regions at days E 20-PO, reaching its mature extension at day P2. After P16 alpha MSH positive threads, possessing varicosities are restricted to the same areas as ACTH 1-39 like fiber positivity is.

Nerve cell bodies and fibers displaying Substance P-like (SP-li) immunoreactivity have been identified in the pedal ganglia (PG) of Mytilus galloprovincialis. Most reactive neurons are small-sized, unipolar and are located in the ganglionic cortex. Positive fibers form a dense network in the neuropile and run parallel in the commissure and in the nerves and connective tracts. The positivity of the labelled elements to a monoclonal antibody indicates that SP-li substance of Mytilus contains a portion immunologically similar to the C-terminal pentapeptide responsible for the pharmacological activity of vertebrate SP.

Chromogranins A and B and secretogranin II have been localized in a wide spectrum of gastroenteropancreatic endocrine/paracrine cells. Chromogranin A immunoreactivity showed the widest distribution and was displayed by glucagon-, PP-, gastrin-, gastrin-CCK-, secretin-immunoreactive cells, the most intense stainings being peculiar of enterochromaffin cells. Chromogranin B immunoreactivity was detected in gastrin- and glucagon cells and in some enterochromaffin cells containing also chromogranin A. Secretogranin II was paired to chromogranin A in glucagon cells of pancreatic islets or occurred alone in glycentin/PP cells of colonic mucosa. Neither of the chromogranins nor secretogranin II have been so far detected in somatostatin-, GIP-, or motilin-immunoreactive cells. Chromogranin A but not chromogranin B or secretogranin II has been detected in the gastric argyrophilic ECL cells.