Isolation of a YAC clone covering a cluster of nine S100 genes on human chromosome 1q21: rationale for a new nomenclature of the S100 calcium-binding protein family

S100 proteins are low-molecular-weight calcium-binding proteins of the EF-hand superfamily and appear to be involved in the regulation of a number of cellular processes such as cell cycle progression and differentiation. More than 10 members of the S100 protein family have been described from human sources so far. We have now isolated a YAC clone from human chromosome 1q21, on which 9 different genes coding for S100 calcium-binding proteins could be localized. Moreover, we have mapped the gene coding for S100P to human chromosome 4p16 and thereby completed the chromosomal assignments of all known human S100 genes. The clustered organization of S100 genes in the 1q21 region allows us to introduce a new logical nomenclature for these genes, which is based on the physical arrangement on the chromosome. The new nomenclature should facilitate and further the understanding of this protein family and be easily expandable to other species.     

Glucose as an auxiliary substrate

Summary A theoretical consideration is presented of the comparative efficiency of carbon conversion of glucose by the Embden-Meyerhof-Parnas (EMP) and the oxidative hexosemonophosphate (HMP) pathways. As a result it is shown that maximum carbon conversion, that is 89%, is possible when glucose is assimilated via the EMP pathway. This value is diminished in proportion to the participation of the HMP pathway in carbon assimilation and is halved when glucose is incorporated entirely via this pathway. If NADPH is included as a source of energy, glucose may behave both as an excess carbon and an excess energy substrate, the latter being the case when greater portions of the HMP pathway operate, and the extent of this is in turn dependent on the P/O quotient. If NADPH cannot be used for ATP synthesis, glucose remains an excess carbon substrate throughout, although when the HMP pathway accounts for more than 26% of glucose assimilation an increasing excess of reduction equivalents is produced. These results are interpreted in terms of mixed-substrate utilization for improving growth yield when glucose is to be used as the excess carbon component.     

Identification of Merkel cells in human skin by specific cytokeratin antibodies:

Merkel cells are special neurosecretory cells which, in adult human skin, are usually very scarce. By immunofluorescence microscopy using antibodies to human cytokeratin polypeptide no. 18, we localized distinct non-keratinocyte cells in the glandular ridges of human fetal and adult plantar epidermis. Using electron and immunofluorescence microscopy, these cells were identified as Merkel cells containing typical neurosecretory granules as well as bundles of intermediate-sized filaments and desmosomes. Two-dimensional gel electrophoresis of the cytoskeletal fractions of microdissected epidermal preparations highly enriched in Merkel cells indicated the presence of cytokeratin polypeptides nos. 8, 18 and 19 which are typical of diverse simple epithelia of the human body. Double immunofluorescence microscopy showed that these human Merkel cells contain neither neurofilaments nor vimentin filaments. In human fetuses of 18-24 weeks of age, conspicuously high concentrations of Merkel cells, reaching a density of approximately 1,700 Merkel cells/mm2 skin, were found in the glandular ridges of plantar skin. The concentration decreased considerably at newborn and adult stages. Thin cell processes (up to 20 microns long) were observed in many fetal epidermal Merkel cells. In addition, we detected isolated Merkel cells deeper in the dermis (i.e. at distances of, at most, 100 microns from the epidermis) in fetal and newborn plantar skin. Our results show that Merkel cells are true epithelial cells which, however, differ profoundly from epidermal keratinocytes in their cytokeratin expression. The findings are discussed in relation to the much disputed question of the origin of Merkel cells. The present data speak against the immigration of Merkel cells from the neural crest, but rather suggest that they originate from epithelial cells of the skin, although most probably not from differentiated keratinocytes.     

Peroxidases in Acetabularia: their possible role in development

Abstract. Crude enzymatic extracts from Acetabularia exhibit very low peroxidase activity after a lag period. Starch gel electrophoresis of extracts from growing algae shows a single, extremely anodic band. Extracts of small, slow-growing or cap-bearing algae, which do not grow any more, do not exhibit any peroxidase band. Cytochemical staining with benzidine reveals changes in both the quantity and distribution of peroxidase along the polarized Acetabularia cell. The homogenous staining of small algae becomes distributed along a negative apico-basal gradient when the algae initiate their rapid growth phase. This polarized pattern is repeated on the hair whorls. A similar developmental sequence directs cap growth, with an initial intense staining reaction of the primordium, which later leaves only the corona inferior stained blue. Finally, the Acetabularia cell remains slightly blue at the edges of the rhizoidal out-growths and cap rays. Crude extracts of Acetabularia induce a lag in standard horseradish peroxidase (HRP) activity. The inhibitor is always present in small and growing algae; it is sometimes absent or less active in cap-bearing algae. In no case does it change the kinetics of the HRP reaction with guaïacol. The lag is completely suppressed by pretreatment with either H₂O₂ or ascorbate oxidase. The changes in peroxidase activity, correlated with developmental stage and according to a polarized gradient, suggest that the enzyme could be involved in some way in the control of morphogenesis in Acetabularia . An inhibitor of peroxidase activity, which disappears as the cap matures, might, in turn, exert a regulatory function.     

Effect of uracil on colonial morphology ofNeisseria gonorrhoeae

The production of cells ofNeisseria gonorrhoeae that form type-T4 colonies in cultures started with cells that originally formed only type-T2 colonies was inhibited by calf thymus RNA. Guanosine and uracil were the only nucleic acid constituents that significantly reduced the T2 to T4 shift. Uracil gave the best results in degree of inhibition. It was found that some tritiated uracil was incorporated into the RNA of growing cells ofN. gonorrhoeae but that much more was incorporated into DNA probably after conversion to guanine and adenine. The data show that the shift from T2 to T4 can be progressively inhibited by increasing the concentration of uracil in the growth medium.     

Changes in the surface morphology of the rabbit endometrium related to the estrous and progestational stages of the reproductive cycle

Summary Changes occurring on the surface of the uterine luminal epithelium of the rabbit during the estrous and progestational stages of the reproductive cycle were examined by scanning and transmission electron microscopy. The findings demonstrate that the uterine epithelium, or endometrium, contains two cell types: ciliated cells and nonciliated, microvillous cells. In estrous animals, ciliated cells, although not very numerous, were usually observed in small groups. However, at increasing intervals of time following mating, ciliated cells progressively disappeared from the endometrium until approximately eight to ten days post coitum, when they became scare. From several hours to four to five days following mating, extensive changes occurred on the surfaces of microvillous cells. When observed by TEM, these elements contained organelles typical of cells involved in the synthesis and secretion of glycoproteins. Furthermore, microvillous cells during this period displayed numerous apical protrusions of different sizes and shapes and containing material of varying electron density. Parallel SEM examinations of the same material confirmed the presence of these protrusions. Some of the protrusions appeared as spheroidal masses attached to the cytoplasm by means of a cytoplasmic strand. Other surface masses were clearly unattached to the cell surface and were distributed (1) on the surface of microvillous cells, (2) on the cilia of adjacent ciliated cells, and (3) on the surface of spermatozoa.Changes occurring on the luminal surface during the early postcoital period are interpreted as an expression of morphodynamic processes likely involving coupled secretion (exocytosis) and resorption (endocytosis) of luminal material. The observations presented here also demonstrate that between six and ten days post coitum, the rabbit endometrium contained increasing numbers of enlarged, nonciliated cells that probably arose by the fusion of smaller, microvillous elements.The work reported here was supported by C.N.R. contracts No. CT 760128809 and CT 77014239 (to P.M.) and NIH. Grant HD-04274 (to J.V.B.)     

SIEVE-ELEMENT ONTOGENY IN THE ROOT OF EQUISETUM HYEMALE

Roots of Equisetum hyemale L. var. affine (Engelm.) A. A. Eat. were fixed in glutaraldehyde, postfixed in osmium tetroxide, and sieve elements of various ages were examined with the electron microscope. Young sieve elements are distinguished by their position within the vascular cylinder and by the presence of numerous refractive spherules, which originate within dilated portions of the endoplasmic reticulum (ER). Early in development, the sieve-element walls undergo a substantial increase in thickness. This is followed by the appearance of massive ER aggregates in the cytoplasm and then by a phase involving stacking and sequestering of the remaining ER. Nuclear degeneration is initiated shortly after the appearance of the ER aggregates. The chromatin condenses into masses of variable size along the inner surface of the nuclear envelope. The envelope then ruptures and chromatin is released into the cytoplasm. During the period of nuclear degeneration, mitochondria and plastids undergo structural modification, while components such as dictyosomes, microtubules, and ribosomes degenerate and disappear. The remaining cytoplasmic components assume a parietal position in the cell, leaving the lumen of the cell clear in appearance. At maturity, the plasmalemma-lined sieve element contains plastids, mitochondria, some ER, and refractive spherules. At this time many of the refractive spherules are discharged into the region of the wall. Pores between sieve elements occur largely on the end walls. During pore development, tubules of ER apparently traverse the pores, but because of the presence of massive callose deposits in the material examined, the true condition of mature pores could not be determined. The connections between mature sieve elements and pericycle cells are characterized by the presence of massive wall thickenings on the pericycle-cell side. Plasmodesmata in the wall thickening are matched by pores on the sieve-element side. Ontogenetic and cytoplasmic factors argue against use of the term “companion cell” for the vascular parenchyma cells associated with the sieve elements.     

Flavone 5-O-glucosides from Daphne sericea

The aerial parts of Daphne sericea yielded two new flavonoids, luteolin 7-methyl ether 5-β-d-glucoside and luteolin 7,3′-dimethyl ether 5-β-d-glucoside, as well as luteolin 7-methyl ether, isovitexin, apigenin and its 7-β-d-glucoside.     

Phospholipid metabolism in the brown alga, Fucus serratus

The metabolism of phospholipids in the brown alga, Fucus serratus was studied. The major phospholipids of this alga are phosphatidylethanolamine, phosphatidylglycerol, phosphatidylinositol, cardiolipin and phosphatidylcholine. When the time-course of labelling of the lipids from [³²P] orthophosphate was studied, total labelling was approximately linear for 8 hr. All the major classes of phospholipid were labelled. The extent and pattern of labelling were not affected by the presence of proteins synthesis inhibitors phosphatidic acid was highly labelled at short time intervals. Phosphatidylcholine was relatively poorly labelled. The extent and pattern of labelling were not affected by the presence of protein synthesis inhibitors indicating that the enzymes involved in phospholipid synthesis have a rather slow turnover. Incorporation of radioactivity into phosphatidylglycerol was stimulated significantly by light.