VIP36, a novel component of glycolipid rafts and exocytic carrier vesicles in epithelial cells.

In simple epithelial cells, apical and basolateral proteins and lipids in transit to the cell surface are sorted in the trans-Golgi network. We have recently isolated detergent-insoluble complexes from Madin-Darby canine kidney cells that are enriched in glycosphingolipids, apical cargo and a subset of the proteins of the exocytic carrier vesicles. The vesicular proteins are thought to be involved in protein sorting and include VIP21-caveolin. The vesicular protein VIP36 (36 kDa vesicular integral membrane protein) has been purified from a CHAPS-insoluble residue and a cDNA encoding VIP36 has been isolated. The N-terminal 31 kDa luminal/exoplasmic domain of the encoded protein shows homology to leguminous plant lectins. The transiently expressed protein is localized to the Golgi apparatus, endosomal and vesicular structures and the plasma membrane, as predicted for a protein involved in transport between the Golgi and the cell surface. It is diffusely localized on the plasma membrane but can be redistributed by antibody modulation into caveolae and clathrin-coated pits. We speculate that VIP36 binds to sugar residues of glycosphingolipids and/or glycosylphosphatidyl-inositol anchors and might provide a link between the extracellular/luminal face of glycolipid rafts and the cytoplasmic protein segregation machinery.     

Reproductive activity of ruffed lemurs (Varecia variegata variegata) in a Madagascar rain forest

Mating activity was observed during four breeding seasons in two groups of black and white ruffed lemurs (Varecia variegata variegata) living in lowland rain forest on Nosy Mangabe island, Madagascar. The onset of the May-July breeding season was signalled by behavioral changes in adult males. Males made forays outside their usual home ranges, were more aggressive to other males, and performed appetitive and other sex-specific behaviors more frequently. Females showed receptive and proceptive behaviors during a 1-2 day behavioral estrus. Ruffed lemurs mated monogamously, polyandrously, and polygynously. These observations do not support previous assertions that they live only in monogamous families. Limited evidence suggests females exercised mate choice and may have preferred familiar males. © 1993 Wiley-Liss, Inc.     

Subtypes of non-transformed human mammary epithelial cells cultured in vitro: histo-blood group antigen H type 2 defines basal cell-derived cells

Abstract. Normal (non-transformed) human mammary epithelial cell lines derived from reduction mammoplasties were analyzed by immunocytochemistry with more than 80 monoclonal antibodies (mAbs) and other specific reagents to tissue-specific and developmentally regulated antigens at different passage levels. A subpopulation of poorly differentiated, proliferating epithelial cells, corresponding to the 'selected' cell type of late passages, is shown to be characterized by a new marker, the histo-blood group antigen H type 2, probably carried on a membrane-bound glycolipid. These cells also express a number of other onco-developmental carbohydrate antigens [Le^y, Le^x, sialosyl-Le^a, precursor of Thomsen Friedenreich antigen (T_n), but not Thomsen-Friedenreich antigen and sialosyl-T_n]. Their cytokeratin (CK) phenotype, as assessed by reactivity with monospecific mAbs and two-dimensional gel electrophoresis, is CK 5, 6, 14 and 17, with CK 19 being consistently absent, and varying minor amounts of CK 7, 8 and 18, as well as 15 and 16. The reactivity of these cells with a panel of 11 mAbs specific for CK 18 varies considerably even after cloning, indicating heterogeneity of epitope expression or accessibility. Our data strongly suggest that the H type 2⁺ cells develop from the basal cell layer of the mammary gland.     

Shoulder muscle load and muscle fatigue among industrial sewing-machine operators

Physiological responses to physical work were assessed for 29 female industrial sewing-machine operators during an 8-h working day under ordinary working conditions. During sewing-machine work, the average (left and right) static load in the trapezius muscle was 9% of the maximal electromyogram (EMG) amplitude (% EMG_max), while the average mean load was 15% EMG_max, and the average peak load was 23% EMG_max. The static load level was unrelated to the muscle strength of the sewing-machine operators, which for the group as a whole was within the normal range. The load levels remained unchanged during the working day, while changes in the EMG mean power frequency and zero crossing frequency rate occurred, both indicating the development of muscle fatigue in left and right trapezius muscle during the working day. In line with this, the rating of perceived exertion in the shoulder and neck region increased during. the working day. Dividing the group of sewing-machine operators into two groups, those with the highest frequency and those with the lowest frequency of shoulder/neck troubles showed that the former group had significantly lower muscle strength, despite the fact that no differences in the surface EMG during sewing were found between the two groups. It was concluded that industrial sewing-machine work involves a pattern of shoulder muscle activity which induces fatiguing processes in the shoulder and neck regions. Furthermore, since the static shoulder muscle load was independent of muscle strength, factors other than working posture may be of significance for the static shoulder muscle load.     

Ethnobotany of Atlantic Forest coastal communities: Diversity of plant uses in Gamboa (Itacuruçá island,Brazil)

Local plants are a very important resource for the community of Gamboa, located at Itacuruçá Island, Sepetiba Bay, State of Rio de Janeiro, Brazil Ninety species of plants, belonging to 40 families, are used for a variety of purposes, such as food, construction, handicraft, and medicine. In a survey medicinal uses for plants were the most quoted by the community. Uses of medicinal plants within Gamboa and with other coastal communities are analyzed using diversity indices. Use by different categories of people based on sex, age, and economic activity was compared and significant differences were found among the groups compared, except for economic categories (fishermen and non-fishermen). The theory of island biogeography is shown to be useful for analyzing different levels of resource uses on different islands.     

The effects of terminal heterologies on gene targeting by insertion vectors in embryonic stem cells.

We have examined the effects of placing nonhomologous DNA on the ends of an insertion-type gene targeting vector. The presence of terminal heterologies was found to be compatible with insertion targeting, and the terminal heterologies were efficiently removed. Terminal heterologies reduced the frequency of gene targeting to variable extents. The degree of inhibition of targeting was dependent on the length and the position of the heterology: 2.1kb heterologous sequences were more inhibitory than shorter regions of heterology, and heterology placed on the end of the long (4.8kb) arm of homology was more inhibitory than heterology positioned on the end of the short (0.8kb) arm. When heterology was placed on both arms of the targeting vector the targeting efficiencies were similar to or higher than when heterology was present on the long arm only. These results suggest that terminal sequences are removed simultaneously from both ends of targeting vectors. The removal of terminal sequences probably occurs by exonucleolytic degradation of both strands at each end, and removal of at least one of the strands is intimately coupled with the process of homologous recombination. These findings have implications for the design of gene targeting vectors.     

Characterization of hemocytes from the marine amphipod Parhyale hawaiensis (Dana 1853): Setting the basis for immunotoxicological studies

Hemocytes are circulating blood cells that play a crucial function in amphipods and other crustacean immune systems. The hemocytes of the marine tropical amphipod Parhyale hawaiensis have been used for the evaluation of DNA damage and micronuclei, but they have not been characterized in the scientific literature. The aim of this study was to describe the hemolymph cells of P. hawaiensis and study their phagocytotic activity. Basic dyes were used to differentiate the cell types and the presence of lipids. The total hemocyte counts (THCs) and the proportion and sizes of the hemocyte types were determined. Hemolymph was exposed to Escherichia coli for verification of the presence of phagocytosis. Three cell types, all containing lipids, were identified in P. hawaiensis: granulocytes (oval shape, 13.4 × 7.6 μm), semi-granulocytes (oval shape, 14.1 × 7.2 μm), and hyalinocytes (round shape, 9.6 × 7.2 μm). Those three cell types were found in different percentages in males (64.8%, 31.1%, and 4.2%) and females (70.1%, 28.2%, and 1.7%). THCs for males were 9007 ± 3800 cells per individual and 4695 ± 1892 cells per individual for females. The cells of E. coli were phagocytized by the hemocytes. Our findings increased the knowledge of hemocytes in P. hawaiensis and is a step forward in using hemocyte-based immune responses as an endpoint in ecotoxicology.     

Functional analysis of the Lactococcus lactis usp45 secretion signal in the secretion of a homologous proteinase and a heterologous α-amylase

The ups45 gene encodes the major extracellular protein from Lactococcus lactis. The deduced sequence of the 27 residue leader peptide revealed the tripartite characteristics of a signal peptide. This leader peptide directed the efficient secretion of the homologous proteinase (PrtP) in L. lactis, indicating that the putative signal peptide of PrtP can be replaced by the 27 residue Usp45 leader peptide. In addition, the 27 residue leader peptide could be used to secrete the Bacillus stearothermophilus α-amylase, encoded by the amyS gene. Fusion of the usp45 promoter region and various parts of the leader sequence to an amyS gene devoid of its signal sequence, showed that in Escherichia coli the first 19, 20, and 27 residues of the Usp45 leader are able to direct α-amylase secretion. In L. lactis the shorter signal peptides did not result in secretion of α-amylase, providing experimental evidence for the hypothesis that gram-positive bacteria require a longer signal peptide for secretion than gram-negative organisms.     

The effect of multiplicity of infection on recombination values in bacteriophageT 4 D

Zusammenfassung In Massenlysaten und Einzelwürfen vonT 4-Kreuzungen steigt der Prozentsatz der Rekombinanten mit steigender Infektionsmultiplizität an, und zwar sowohl zwischen gekoppelten als auch zwischen ungekoppelten Genen. Dieser Befund stimmt mitTrautners (1960) Ergebnissen beiT 1-Kreuzungen überein. Der scheinbarte Widerspruch zuEpsteins (1958) früheren Resultaten beiT 4-Kreuzungen läßt sich erklären, da verschiedene Wirtsbakterien verwendet wurden.Es wird diskutiert, ob dieser Effekt durch die Annahme erklärt werden kann, daß die infizierenden Phagegenome bei der Rekombination brechen und daß die Bruchstellen zufallsgemäß über die Genome einer Phagenpopulation verteilt sind. Die beschriebenen Versuche schließen jedochTrautners Interpretation nicht aus, daß in der infizierten Bakterienzelle eine Topographie existiert, d. h., daß die verschiedenen Genotypen während der Vermehrung nach Einzelinfektion nicht so vollständig durchmischt werden wie nach Mehrfachinfektion.

---

---

With 2 Figures in the Text