Enzyme histochemistry of human brain tumors and their tissue cultures with special reference to the oxidoreductases in the glioblastoma multiforme

Summary Fresh tissue and tissue cultures of 80 glioblastoma multiforme and 12 monstrocellular sarcoma were histochemically investigated. The activity of the following enzymes was demonstrated in the biopsies and tissue cultures of every tumor: NADH-tetrazolium reductase, NADPH-tetrazolium reductase, lactic dehydrogenase, succinic dehydrogenase, glutamic acid dehydrogenase and cytochrome oxidase. No major differences in the relative activity pattern was shown when fresh tissue and tissue cultures were compared, nor did the enzymatic pattern change during the four week observation time.In both groups major quantitative differences in the enzymatic activity of the tumor cells in the same tissue area or tissue cultures were frequently a striking finding. Differences in the intracellular localization of the enzymatic activity were also observed. In slowly growing gliomas these histochemical variations are absent.     

Distribution of LHRH in the rat and mouse brain with special reference to the tanycytes

Summary The distribution of luteinizing hormone-releasing hormone (LHRH) was studied in the rat and mouse brain by means of light and electron microscopic immunohistochemistry using the peroxidase-antiperoxidase method. An immunoreactive product to LHRH antiserum was found near the blood vessels of the vascular organ of the lamina terminalis. In the arcuate nucleus-median eminence region, an immunoreactive material occurred bilaterally in the hypothalamic tissue around the tuberoinfundibular sulci. Electron microscopy revealed that immunoreactive fibers observed light microscopically contain numerous granules 100–130 nm in diameter. No immunoreactive product was located in the tanycytes of the median eminence, the perikarya of hypothalamic neurons, and the parenchyma of several circumventricular organs (subfornical organ, subcommissural organ, pineal organ, area postrema).Supported by grants from the Ministry of Education of Japan and the Ford Foundation     

Current usage of nomenclature for parasitic diseases, with special reference to those involving arthropods

Terminological confusion has been aggravated by efforts to develop a standardized nomenclature for parasitic diseases (SNOPAD) arising from the proposal by Kassai et al., 1988) for a standardized nomenclature of animal diseases (SNOAPAD). To restabilize international nomenclature of parasitic diseases it is recommended that, whenever appropriate, names should follow the 'International Nomenclature of Diseases' (IND) compiled by the Council for International Organizations for Medical Sciences (CIOMS/WHO, 1987). For diseases not included in IND, familiarity should guide the choice of name: traditional English language names of diseases should be preferred, e.g. 'malaria', 'scabies' or, for parasitic diseases having no traditional name, the taxonomic name of the causative organism should be applied, e.g. 'Brugia timori microfilaraemia'; 'Plasmodium malariae infection'; 'Simulium allergy'--instead of the generic derivatives proposed by SNOPAD, i.e. brugiosis, plasmodiosis and simuliidosis, respectively. For names of new diseases or those rarely mentioned, the suffix -osis would normally take precedence. Generally, the name of choice for any disease in any language should be the vernacular term, with commonest English usage preferred for international communication, and publications should include synonyms in the list of keywords.     

Enzyme histochemistry of human melanomas and pigmented naevi with special reference to alpha-D-mannosidase activity

Summary A histochemical study of-d-mannosidase revealed that normal human melanocytes (resting state, activated, lentigo simplex) exhibit either no or just detectable activity, as do melanocytes in the initial phase of lentigo maligna. Junctional, or occasionally zone A naevocytes displayed a very low enzyme activity. On the other hand, melanocytes in the initial stage of neoplastic transformation (dysplastic naevi, advanced stage of lentigo maligna) and also melanoma cells in disorders of low malignant potential (initial naevogenic melanoma, superficial spreading melanoma) displayed a high activity uniformly throughout the cell population. In the malignant forms (nodular melanoma, recurrences, metastases), the enzyme activity was remarkably heterogeneous, suggesting a breakdown of uniformity during malignant transformation. The significance of -mannosidase activity induction in the course of melanocyte neoplastic transformation is not clear at present. The results of biochemical assays suggest that the lysosomal isoenzyme is mainly responsible. Other lysosomal enzymes, and dehydrogenases studied concomitantly, did not display any comparable phenomena of induction or similar behaviour. However, the results of a comparison of-mannosidase with the melanocyte reference enzyme tyrosinase suggested activity patterns in the enzyme pair which may provide a better insight into the biochemical differentiation of human melanocytes in neoplastic disorders. The possible relationship of-mannosidase to melanogenesis is also discussed.     

The specific heat of red cells,with special reference to the paracrystalline rat red cell

The specific heat of the rat red cell, kept in cold sodium citrate, changes in the neighborhood of 6°C., the temperature near which the cell passes from its paracrystalline state to a state of greater disorder. The change in the specific heat is from 0.74 with a standard deviation of ±0.022 (paracrystalline state) to 0.87 with a standard deviation of ±0.021 (normal state). Although it has been looked for, no evidence of a change in specific heat has been found, between 1°C. and 15°C., in the case of the human red cell or of the fresh rat red cell in saline or plasma.     

Model film studies in enzyme histochemistry with special reference to glucose-6-phosphate dehydrogenase

Summary This paper deals with the progress made over the last few years in our understanding of enzyme cytochemical staining methods as studied using a fundamental approach with the aid of a model system of thin gel films. Although model films with a matrix of polyacrylamide have been mostly used, the properties and possible applications of other matrices are also reviewed. The chemical aspects of the entrapment of enzyme molecules into a matrix are summarized. Special attention has been paid in model film studies to the principles of the trapping reaction of a diffusable precursor resulting from the enzymatic conversion of a substrate. They are considered here as they concern the cytochemical demonstration of acid phosphatase activity with a lead salt. The effect of fixatives on different enzyme activities, the diffusion rate of substrates and chromogenic compounds to the enzyme site, and enzyme kinetics under cytochemical conditions are also discussed, since they are factors which influence the final results of the staining procedures. The advantage of model film studies in enabling the direct correlation of cytochemical and biochemical results is outlined with special reference to the cytochemical determination of glucose-6-phosphate dehydrogenase with Tetra Nitro BT. A method for determining enzyme activities in the soluble fraction of isolated cells after incorporation in model films is described for the first time. This method has proved to be highly appropriate for microscopical observations of glucose-6-phosphate dehydrogenase activity in single cells, because it results in a good morphology and no formazan precipitaties outside the cells. On the other hand, this type of model film forms a bridge between fundamental model film studies using purified enzyme and quantitative enzyme cytochemistry performedin situ.     

The postnatal development of the rat kidney,with special reference to the chemodifferentiation of the proximal tubule

Summary New nephron anlages appear in the renal cortex up to the 4th postnatal day (PD). The last anlages to be formed develop into functional nephrons by PD 10, and the cortex appears mature at PD 12 after formation of the cortex corticis. The renal medulla develops by the longitudinal growth of loops of Henle and collecting ducts. The immature medulla cannot be divided into different zones and corresponds structurally to the later inner stripe of the outer zone. The inner zone is formed by PD 8, and the outer stripe of the outer zone by PD 12. The renal medulla is mature at PD 21.From the start of its development, the renal proximal tubule consists of the pars convoluta and pars recta. In both parts the formation of the brush border is accompanied by the simultaneous appearance of brush border enzymes (alkaline phosphatase, -glutamyltranspeptidase, dipeptidylamino-peptidase IV) and lysosomal enzymes (acid phosphatase, acid -galactosidase, N-acetylglucosaminidase, dipeptidylaminopeptidase II) over the full length of the proximal tubule. During the course of proximal tubule maturation, however, the lysosomal enzyme activities decline in the pars convoluta (with constant brush border enzyme activities), while the brush border enzyme activities increase in the pars recta (with constant lysosomal enzyme activities). The two parts further differ in that they exhibit different lysosomal patterns from the outset, the pars convoluta containing numerous large, highly enzyme-active lysosomes arranged in groups, and the pars recta containing only a few very small lysosomes with low enzyme activity. Thus, even in the newborn rat, the lysosomal pattern of the pars recta already corresponds to that of the mature S₃ segment. The S₁ and S₂ segments of the pars convoluta first differentiate between PD 10 and 21, as the groups of large lysosomes are progressively broken up and the extent of the lysosomal apparatus is diminished, this proceeding in a retrograde direction from the end of the immature pars convoluta.Supported by the Deutsche Forschungsgemeinschaft (SFB 105)     

Cytological studies of developing muscle with special reference to myofibrils,mitochondria, Golgi material and nuclei

Summary Developing striated muscle in the chick embryo was studied with special reference to the cytological changes which occur as the myofibrils are being elaborated. Observations were made on the mitochondria, cytoplasmic granules, Golgi material, neutral red bodies, nuclei, and nucleoli, and on the developing fibrils. Particular attention was directed to the early part of the histogenetic process when a few primary myofibrils are being formed and becoming striated. This period is characterized by hypertrophy of the Golgi material, nuclei, and nucleoli.The myofibrils were found to arise in close association with filamentous mitochondria, apparently at the expense of the numerous small cytoplasmic granules which fill the early myoblasts. The mitochondria change staining reaction during this time and to a certain extent disappear. Many ring-shaped mitochondria, the significance of which is not known, were found among the unchanged mitochondria which remain near the nucleus.At first the myofibrils are homogeneous and do not take the stain well. Later they stain heavily, although they remain homogeneous, and finally striation appears. Evidence was presented that the Z-membrane begins development before striation is visible.When the primary fibrils have become striated, formation of myofibrils by the above method apparently ceases and further increase in the number of fibrils is probably brought about by longitudinal splitting of those formed first.At this time, also, the Golgi material decreases in amount, the nuclei become smaller, and the nucleoli break up and lose their distinct form.A secondary fibril system was also described which, it is believed, is distinct from the myofibril system.Mitotic division followed by cleavage of the cytoplasm occurs in the earliest myoblasts, probably for the purpose of increasing the number of these cells. However, as soon as fibrillization sets in, mitotic division ceases and the nuclei multiply by amitosis not followed by cleavage of the cytoplasm.The nucleoli divide before amitotic division of the nuclei. Evidence was presented that certain nuclear granules may be associated with the amitotic division process.The Golgi material in embryonic muscle is situated at the two poles of the nuclei. This furnishes evidence in favor of the belief that the Golgi material of adult muscle has a similar location and is not represented by the Cajal-Fusari network.Neutral red granules were demonstrated near the nucleus and in the axial cytoplasm, but they did not coincide in position with the Golgi material.It was noted that the cytological changes which take place when the myofibrils are being elaborated in the developing myoblasts resemble those in gland cells at the time secretory materials are being formed.