Metabolism of 15-hydroxyeicosatetraenoic acid by Caco-2 cells

Monolayers of Caco-2 cells, a human enterocyte cell line, were incubated with [1-14C]15-hydroxyeicosatetraenoic acid (15-HETE), a lipid mediator of inflammation, and [1-14C]arachidonic acid. Both fatty acids were taken up readily and metabolized by Caco-2 cells. [1-14C]Arachidonic acid was directly esterified in cellular phospholipids and, to a lesser extent, in triglycerides. When [1-14C]15-hydroxyeicosatetraenoic acid was incubated with Caco-2 cells, about 10% was directly esterified into cellular lipids but most (55%) was beta-oxidized to ketone bodies, CO2, and acetate, with very little accumulation of shorter carbon chain products of partial beta-oxidation. The radiolabeled acetate generated from beta-oxidation of [1-14C]15-hydroxyeicosatetraenoic acid was incorporated into the synthesis of new fatty acids, primarily [14C]palmitate, which in turn was esterified into cellular phospholipids, with lesser amounts in triglycerides. Caco-2 cells were also incubated with [5,6,8,9,11,12,14,15-3H]15-hydroxyeicosatetraenoic acid; most of the radiolabel was recovered either in ketone bodies or in [3H]palmitate esterified in phospholipids and triglycerides, demonstrating that most of the [3H]15-hydroxyeicosatetraenoic acid underwent several cycles of beta-oxidation. The binding of both 15-hydroxyeicosatetraenoic acid and arachidonic acid to hepatic fatty acid binding protein, the only fatty acid binding protein in Caco-2 cells, was measured. The Kd (6.0 microM) for 15-HETE was three-fold higher than that for arachidonate (2.1 microM).     

Radiocarbon age of carbonate in fruits of Lithospermum from the early Bronze Age settlement of Hirbet ez-Zeraqōn (Jordan)

Uncarbonized fruits of the Boraginaceae occur widely in cultural layers of archaeological sites in the Mediterranean and the Near East. To date, interpreting their origin remains problematic. It is difficult for archaeobotanists to tell whether such fruits were deposited as part of the cultural layer or were introduced post-depositionally. In an attempt to answer this question for the early Bronze Age site of Hirbet ez-Zeraqon (Jordan), we used direct ¹⁴C dating of biogenic carbonate from calcareous fruits of Lithospermum. The radiocarbon ages of seven fossil samples of the fruits suggest that they originate from plants that grew during or around the time of occupation.     

The biology of the spiny icefish <Emphasis Type="Italic">Chaenodraco wilsoni</Emphasis> Regan, 1914

The most abundant ice fish species observed in catches off the northern tip of the Antarctic Peninsula in the last 25–30 years has been the spiny ice fish Chaenodraco wilsoni Regan 1914. C. wilsoni has been exploited on a commercial scale from the late 1970s to the end of the 1980s off Joinville–D’Urville Islands (CCAMLR Statistical Subarea 48.1) and in the Cosmonauts and Cooperation Seas and Prydz Bay in the Indian Ocean sector (CCAMLR Statistical Division 58.4.2). This paper presents new information on biological features and life history characteristics of C. wilsoni, based on research survey collections along the northern Antarctic Peninsula in 2006 and 2007 and samples taken in the commercial fishery in 1987. Length frequency compositions from the research surveys demonstrated that fish 21–34 cm long predominated in the catches. Sexual maturity is attained at 24–25 cm. Absolute fecundity and relative fecundity is low (1,000–2,500 eggs; 6–12 eggs). Oocyte diameter varied from 4.0 to 4.9 mm very close to spawning. Spawning at the tip of the Antarctic Peninsula is likely to occur in October–November. Remotely operated vehicle deployments in the northern Weddell Sea demonstrated that C. wilsoni exhibit parental nest guarding where males protect the eggs. The incubation period is likely to be 8 months long. Fish feed primarily on Antarctic krill (Euphausia superba) in the Antarctic Peninsula region and in the Cosmonauts and Cooperation Seas while fish take ice krill (Euphausia crystallorophias), Pleuragramma antarcticum and myctophids to some extent in other areas. Age determination still awaits validation. Preliminary ageing attempts suggested a maximum age of about 8–10 years.     

TNFR1 mediates the radioprotective effects of lipopolysaccharide in the mouse intestine

LPS is radioprotective in the mouse small intestine through a mechanism that includes the synthesis of cyclooxygenase-2 (COX-2) and PGE2. The goal of this study was to identify the intermediate steps in this process. We used wild-type (WT) C57BL/6 mice and knockouts for tumor necrosis factor receptors 1 and 2 (TNFR1-/-, TNFR2-/-) and recombination-activating gene 1-/- mice. Mice were given parenteral LPS and then subjected to 12 Gy total body gamma irradiation. The number of surviving intestinal crypts was assessed 3.5 days after irradiation using a clonogenic assay. Crypt cell apoptosis was assessed by histology. Parenteral administration of LPS induced COX-2 expression, PGE2 production, and radioprotection in WT and TNFR2-/- mice but not in TNFR1-/- mice. TNFR1-/- mice were radioprotected by administration of exogenous 16,16-dimethyl PGE2. Immunohistochemical studies localized TNFR1 and COX-2 expression to subeptihelial fibroblasts and villus epithelial cells. Radiation-induced apoptosis was reduced by pretreatment with LPS in WT and TNFR2-/- mice but not in TNFR1-/- mice. In the absence of LPS, crypt survival was elevated in TNFR1-/- when compared with WT mice. These findings demonstrate that TNFR1 function is required for LPS-induced radioprotection in C57BL/6 mice and define an essential role for TNFR1 function in the induction of COX-2 expression and PGE2 production in this process. The immunolocalization of TNFR1 and COX-2 expression to subepithelial fibroblasts following LPS administration suggests that this cell type plays an intermediate role in LPS-induced radioprotection in the intestine.     

Vegetation development and human occupation in the Damascus region of southwestern Syria from the Late Pleistocene to Holocene

Rockshelter Baaz in the Damascus region of Syria provided a variety of botanical remains from the Late Pleistocene and Early Holocene period. These remains provide new information about the vegetation evolution in this region. The earliest occupational levels correspond with a moisture peak during the Late Pleistocene, between ca. 34–32 kyr b.p., when pine expanded. The next occupations took place during extreme arid conditions, ca. 23–21 kyr b.p., and probably during the Last Glacial Maximum when a steppe vegetation was established. The occupation level of the Younger Dryas, represented by Natufian remains, suggests that the area had been covered by almond-pistachio steppe, similar to later periods of the Early Holocene, and was probably located just outside the range of dense wild cereal stands. There is no drastic impact of the Younger Dryas visible on the vegetation in the botanical remains. The lack of fruits and seeds at Baaz indicates that the site was more likely to have been a temporary hunting post rather than a plant processing site for much of its history. It is ideally suited to this purpose because of its location over the Jaba′deen Pass and the associated springs. However, archaeological remains from the Natufian period, suggest that the site was more permanently occupied during this time. A. Dodonov deceased.     

Nucleic acid binding characteristics of group A/B hnRNP proteins

hnRNP proteins are primarily defined by their specific sedimentational, reconstitutional, and extraction properties and are presumed to be RNA binding. However, it is not clear whether all these proteins have RNA binding capabilities. Recently, using two monoclonal antibodies, fA12 and AC88, we reported that the abundance of a subclass of the highly basic A/B hnRNP proteins was specifically down regulated during terminal differentiation of human and murine cells in vitro. In this report we have examined the nucleic acid binding characteristics of this subclass and other members of the A/B hnRNP proteins in vitro. All members of class A/B hnRNP proteins appear to have similar but not identical nucleic acid binding characteristics. However, the subclass of proteins recognized by AC88 and fA12 exhibit stronger binding affinities and are shown to be highly selective in their binding to RNA vs DNA in vitro. These proteins also preferentially bind poly(U) RNA, suggesting that in vivo they may bind effectively to uridine rich motifs critical in pre-mRNA processing.     

The skull in normal and pugheaded females of the mosquitofish Heterandria formosa Agassiz, 1853 (teleostei, poeciliidae)

Normal and pugheaded skulls in Heterandria formosa were studied by a modified Spalteholz-method, scanning electron microscopy, and X-ray projection microscopy. Pugheadedness was examined for the first time in this fish species. X-ray projection microscopy quickly produces useful results of fish morphology. The modified Spalteholz-method, however, has a higher resolution and single bones and bone sutures can be demonstrated. In pugheaded skulls of Heterandria formosa the neurocranium and upper jaw are shortened, whereas the lower jaw usually remains unaltered. The pugheadedness in H. formosa involves not only a shortening of the vomer, parasphenoid, and maxillaries, but also by displacement and/or curve of the nasals, frontals, vomer, and palatines. For these reasons, the forehead is upraised and steep in pugheads. The origin of pugheadedness is discussed.     

Masculinization in a hermaphroditic female of the mosquitofishHeterandria formosa

A masculinized female ofHeterandria formosa is described for the first time. In size it ranges with 28 mm total length between adult males (17–20mm) and adult females (35–40mm). A masculinized anal fin of about 5 mm length is clearly evident. Rays I and II are reduced as in normal males. Rays III to V are elongated and show features of a developing gonopodium. The remaining rays VI to VIII show the expected shape of a “normal fin.” The genus-specific hook which normally originates in the anterior ramus of ray IV and the proximal serrae which derive from the posterior ramus of ray IV are absent in the masculinized analis. The gravid spots of pregnant normal females are absent. The causes for the formation of a masculinized anal fin in a female ofH. formosa are still unknown. In the masculinized female a hermaphroditic gonad is found in which female and male sections are readily distinguishable. The female regions are dominant. The oocytes of the hermaphroditic gonad are in the stage of vitellogenesis showing a thin zona radiata. The male regions with spermatozoa and Sertoli cells are distributed throughout the ovarian tissue. This mode of hermaphroditism, in which both mature oocytes and mature spermatozoa simultaneously occur in the gonad, is unique among fishes.     

Electron microscopical and autoradiographical studies on the so-called yolk-nuclei in the oocytes of Nemacheilus barbatulus (L.) and Phoxinus phoxinus (L.) (Pisces, Teleostei) (author's transl)

The yolk-nuclei in the oocytes of the teleosts Noemacheilus barbatulus (L.) and Phoxinus phoxinus (L.) were investigated by electron microscopy, autoradiography and ultrahistochemistry. Yolk-nuclei were only found in stage I. They consist of mitochondria, an osmiophilic material and in Noemacheilus partly of dictyosomes. It was demonstrated by marking with [3H]-uridine, that the osmiophilic material contains RNA. The osmiophilic material gets into the juxtanuclear cytoplasm. There it is surrounded by mitochondria. The oocytes of Noemacheilus and Phoxinus contain up to 40 yolk-nuclei, which are only visible by electron microscopy. The yolk-nuclei of both species dissolve in early stage II. Their components disperse in the cytoplasm.