Ultrastructural characterization of two new human endometrial carcinoma cell lines and normal human endometrial epithelial cells cultured on extracellular matrix

Summary Two new lines of human endometrial carcinoma (HEC) cells, one from an adenocarcinoma and one from a highly metastatic serous papillary carcinoma, were established in culture. Structural and morphologic properties of these cells at early passage were compared with those of cultured normal human endometrial epithelial (NHEE) cells. For these studies, cells were grown on a conventional plastic surface or on an extracellular matrix substrate (Matrigel), and examined by transmission electron microscopy and immunofluorescent light microscopy. The HEC cells appeared morphologically similar on plastic and Matrigel, whereas the NHEE cells showed significantly greater epithelial morphologic differentiation on Matrigel than on plastic. On extracellular matrix, the morphologic differences observed between HEC cells and NHEE cells were primarily of an architectural nature, which may be in part explained by differences between NHEE and HEC cells in the arrangement of actin microfilaments and cytokeratin intermediate filaments. Furthermore, HEC cells displayed extensive networks of vimentin intermediate filaments, which were absent from the NHEE cells. These observations support the hypothesis that architectural deregulation is a prominent feature of endometrial carcinoma, and that cytoskeletal alterations may uncouple HEC cell ultrastructural morphology from the influence of extracellular matrix. This research was supported by research grants CA31733, CA45727, and ES07017, from the National Institutes of Health, Bethesda, MD. G. P. S. is a Jefferson Pilot Fellow in Academic Medicine. A preliminary account of this work was presented at the 1988 U.S.-Canadian Academy of Pathology Annual Meeting (Lab. Inves. 58:12a, 1988).     

Sex differences in compatibility of pair-housed adult longtailed macaques

This research was designed to evaluate the effects of same-sex pair housing on the psychological well-being of adult wild-born longtailed macaques (Macaca fascicularis). We studied behavioral compatibility and stress as measured by urinary cortisol excretion in 15 pairs of each sex. Before they were housed together, the pairs were categorized by noncontact pairedpreference testing as preferred, nonpreferred, or randomly assigned partners. Every aspect of data analysis indicated that the success of pairing was strongly related to gender. Whereas 100% of female pairs were compatible, only eight of the 15 male pairs were still together after two weeks, and only five (33%) showed a degree of compatibility resembling that of females. The psychological well-being of virtually all females seemed to be improved during the physical contact paired-housing conditions; they spent more than one-third of the day engaged in social grooming. Paired adult males had much lower interaction rates than adult females. On average, males were initially somewhat stressed by the introduction to a cagemate as indicated by increased urinary cortisol excretion. The noncontact preference testing procedure was no more predictive of pair success than random assignment. For males, the presence of fighting combined with the absence of grooming during the first 90 min opportunity for physical contact (“introduction”) was associated with pair incompatibility, but not to a statistically significant extent. For research protocols permitting social grouping of this species, the social contact requirement of the USDA Animal Welfare Rules usually can be met for adult females by pair housing. For males, pairing with other adult males often is unsuccessful; by our estimates, at least 20% of males cannot be pair-housed with other males. These sex differences in response to same-sex adults are consistent with the known socioecology of macaques. Further research is necessary to determine whether adult males have a lower need for social contact than females, or whether their needs are better met by other types of social contact. © 1994 Wiley-Liss, Inc.     

The Effects of Lithium on a Neuronal in Vitro Orcadian Pacemaker

Previous studies have suggested a causal connection between abnormalities of the circadian system and affective disorders. The effectiveness of lithium or rubidium as a treatment for affective disorders and the ability of lithium or rubidium to influence circadian pacemakers has stimulated research into the mechanism of lithium's action on circadian systems. In this study we used a neuronal in vitro circadian pacemaker preparation, the eye of the mollusc Bulla, to examine the cellular effects of lithium and rubidium. Continuous extracellular LiCl application lengthens the period of the circadian rhythm of the Bulla pacemaker in a concentration-dependent manner. Rubidium was found to be more effective than lithium in period lengthening. Stable phase delays were generated by 2-h pulses of 395 mM LiCl applied extracellularly from zeitgeber time (ZT) 5-7 (mid subjective day). Concomitant continuous application of 16 mM LiCl and light (a depolarizing agent) generated period lengthening substantially greater than the arithmetic sum of the modest period lengthening of each treatment alone. Furthermore, LiCl pulses, applied together with depolarizing extracellular KC1 concentrations, yielded an increasing magnitude of phase delays with increasing KC1 concentration. These data suggest that LiCl acts intracellularly on the circadian pacemaker cells by entering through a voltage-dependent channel, most likely a sodium channel.     

Some comparative properties and localization of porcine jejunal adenylate cyclase.

Cholera toxin is thought to cause intestinal secretion by activating adenylate cyclase and increasing intracellular 3',5'-cyclic AMP concentrations in intestinal mucosa. Cholera toxin causes profuse secretion of fluid into ligated intestinal loops of both pigs and rabbits, but cholera toxin-induced increases in 3',5'-cyclic AMP concentration are much lower in the pig than in the rabbit. Porcine jejunal adenylate cyclase was examined for unusual properties which might account for a lack of 3'-5'-cyclic AMP accumulation after treatment with cholera toxin. The divalent cation requirements, the pH optimum, and the stimulation by fluoride ion were unremarkable. The Km for ATP was 0.11 mM with negative cooperativity indicated by a Hill coefficient of 0.83. Triton X-100 was inhibitory and guanosine diphosphate methylenephosphate stimulated enzyme activity. Adenylate cyclase activity was highest in the basal and lateral membrane fractions of jejunal mucosa and relatively low in brush-border preparations. Pretreatment of pig jejunum with cholera toxin caused a 30-40% activation of the crude and of the partly purified enzyme. A relatively low activation of adenylase cyclase in pig jejunal mucosa, compared with rabbit, may account for the absence of 3',5'-cyclic AMP accumulation after cholera-toxin treatment in the pig.     

Detection of an enzyme activity which cleaves m7 Guanine from m7 GMP in an extract of embryonic chick lens cells

m⁷ Guanine was cleaved from m⁷ GMP by cytoplasmic enzyme activity in an extract prepared from embryonic chick lens cells. The appearance of m⁷-Guanine was proportional tot he time and concentration of extract. m⁷-Guanine inhibited the reaction but neither guanine nor ribose 5-phosphate did. m⁷ Guanine may be derived from m⁷ GpppG mRNA cap by two enzymatic reactions with m⁷ GMP as a product-substrate intermediate.     

Suavitas imbricata gen. et sp. nov., an anatomically preserved seed analogue of putative lycophyte affinities from upper Pennsylvanian marine deposits

A megasporangiate fructification has been discovered in dysoxic, offshore marine shales from Upper Pennsylvanian deposits of northcentral Texas. This specimen, described as Suavitas imbricata gen. et sp. nov., consists of imbricating, helically arranged sporophylls that diverge from a stout axis. The axis is characterized by parenchymatous pith and weakly developed vascular tissue. Sporangia are terminal with a horizontally elongated, apical dehiscence slit. There is one large functional megaspore in each sporangium, and the megaspore wall is constructed of interconnected rodlets. Parsimony analysis indicates that Suavitas may be related to rhizomorphic lycophytes. The unique combination of characters displayed by this species illustrates the central role played by extinct species in characterizing overall patterns of plant relationships, and emphasizes the uniqueness of plant communities from extrabasinal habitats during the late Paleozoic.     

A novel Brucella isolate in association with two cases of stillbirth in non-human primates – first report

Background  Brucellosis is veterinary and human health problem.
Methods  A 13-year-old wild caught multiparous and an 8-year-old colony-born nulliparous baboon had stillbirths in the second trimester of pregnancy. Culture isolates from both postpartum uteruses were characterized using traditional biochemical analysis, PCR, and multilocus sequencing.
Results  The isolates morphologically resembled Brucella although their phenotypic characteristics were not consistent with any currently described species. The isolates represent a novel lineage within the genus with unique alleles, not previously seen in surveys of greater than 300 isolates representing the known diversity of the genus, present at 5/9 loci examined.
Conclusions  The described cases are to the best of our knowledge the first presentation of a naturally acquired Brucella infection in non-human primates associated with stillbirths from the same colony where Brucella seropositivity in the baboons was described 45 years ago. The organism appears to represent a previously undescribed Brucella species.