Blood and extracellular fluid volume in whole body and tissues of the Pacific hagfish, Eptatretus stouti

Whole-body and 20 individual-tissue (51)Cr-RBC (red cell space; RCS) and (99)Tc-diethylenetriaminepentaacetic acid (extracellular space; ECS) spaces were measured in seven unanesthetized Pacific hagfish (Eptatretus stouti). Volume indicators were administered via a dorsal aortic cannula implanted the previous day. Blood samples were collected at 6, 12, 18, and 24 h after injection. Tissues were removed at 24 h and radioactivity was measured; tissue water content (percent of wet weight) was determined by desiccation at 95 degrees C for 48 h. Mixing rates of both indicators were identical and were essentially complete by 12 h, indicating that blood convection is the rate-limiting process. At 24 h, the whole-body RCS was 19.3+/-2.1 mL kg(-1) body weight, and the ECS was 338.5+/-15.2 mL kg(-1) body weight. Blood volume estimated from the 24-h RCS and the mean central hematocrit (14%) was 137.9 mL kg(-1) body weight. Liver RCS (118.6+/-30.5 microL g(-1) tissue weight) was twice that of any other tissue and was also the most variable, ranging from 59 to 263 microL g(-1), whereas liver ECS (406.0+/-34.3 microL g(-1)) was in the range of other tissues, and water content (66.9%+/-3.5%) was low. Gill RCS (55.9+/-5.7 microL g(-1)), ECS (415.3+/-37.7 microL g(-1)), and percent water (83.1%+/-0.8%) were higher than most other tissues. RCS, ECS, and percent water were consistently lowest in ovum (1.1+/-0.02 microL g(-1), 111.1+/-4.3 microL g(-1), 51.3%+/-3.5%, respectively). Tongue, notocord, and myotome had generally lower RCS (2.1+/-0.4, 2.2+/-0.5, 7.1+/-0.1 microL g(-1), respectively) and ECS (121.2+/-7.0, 246.3+/-17.4, 185.3+/-16.7 microL g(-1), respectively), although their water content was in the midrange (74.7+/-0.5, 81.2+/-1.6, 74.4%+/-0.6%, respectively). Skin had a low RCS (6.8+/-1.1) and midrange ECS (387.5+/-28.0) but very low water content (61.2%+/-2.1%). These findings confirm that hagfish blood volume is at least twice as large as other fish, whereas our estimate of extracellular fluid volume is larger than previously reported and more in line with the predicted interstitial volume. RCS, ECS, and water content vary, often independently, between tissues, which may perhaps be indicative of specific tissue needs or functions. A distinct spleen is lacking in hagfish, and the liver appears to serve this function by sequestering red cells. To our knowledge, this is the first report of tissue ECS in Myxiniformes.     

Reelin and ApoE receptors cooperate to enhance hippocampal synaptic plasticity and learning

Two apolipoprotein E (apoE) receptors, the very low density lipoprotein (VLDL) receptor and apoE receptor 2 (apoER2), are also receptors for Reelin, a signaling protein that regulates neuronal migration during brain development. In the adult brain, Reelin is expressed by GABA-ergic interneurons, suggesting a potential function as a modulator of neurotransmission. ApoE receptors have been indirectly implicated in memory and neurodegenerative disorders because their ligand, apoE, is genetically associated with Alzheimer disease. We have used knockout mice to investigate the role of Reelin and its receptors in cognition and synaptic plasticity. Mice lacking either the VLDL receptor or the apoER2 show contextual fear conditioning deficits. VLDL receptor-deficient mice also have a moderate defect in long term potentiation (LTP), and apoER2 knockouts have a pronounced one. The perfusion of mouse hippocampal slices with Reelin has no effect on baseline synaptic transmission but significantly enhances LTP in area CA1. This Reelin-dependent augmentation of LTP is abolished in VLDL receptor and apoER2 knockout mice. Our results reveal a role for Reelin in controlling synaptic plasticity in the adult brain and suggest that both of its receptors are necessary for Reelin-dependent enhancement of synaptic transmission in the hippocampus. Thus, the impairment of apoE receptor-dependent neuromodulation may contribute to cognitive impairment and synaptic loss in Alzheimer disease.     

Mapping the heparin-binding site on the 13-14F3 fragment of fibronectin

Fibronectin, a multifunctional glycoprotein of the extracellular matrix, plays a major role in cell adhesion. Various studies have revealed that the human 13th and 14th fibronectin type III domains (labeled (13)F3 and (14)F3 here) contain a heparin-binding site. Mapping of the heparin-binding sites of (13-14)F3, (13)F3, and (14)F3 by NMR chemical shift perturbation, isothermal titration calorimetry, and molecular modeling show that (13)F3 provides the dominant heparin-binding site and that the residues involved are within the first 29 amino acids of (13)F3. Predictions from earlier biochemical and modeling studies as well as the x-ray structure of (12-14)F3 were tested. It was shown that the positively charged residues that project into the solvent from the ABE face of the triple-stranded beta sheet on (13)F3 are involved in binding, but (14)F3 does not appear to contribute significantly to heparin binding.     

An integrated, stacked microlaboratory for biological agent detection with DNA and immunoassays

An integrated, stacked microlaboratory for performing automated electric-field-driven immunoassays and DNA hybridization assays was developed. The stacked microlaboratory was fabricated by orderly laminating several different functional layers (all 76 x 76 mm(2)) including a patterned polyimide layer with a flip-chip bonded CMOS chip, a pressure sensitive acrylic adhesive (PSA) layer with a fluidic cutout, an optically transparent polymethyl methacrylate (PMMA) film, a PSA layer with a via, a patterned polyimide layer with a flip-chip bonded silicon chip, a PSA layer with a fluidic cutout, and a glass cover plate layer. Versatility of the stacked microlaboratory was demonstrated by various automated assays. Escherichia coli bacteria and Alexa-labeled protein toxin staphylococcal enterotoxin B (SEB) were detected by electric-field-driven immunoassays on a single chip with a specific-to-nonspecific signal ratios of 4.2:1 and 3.0:1, respectively. Furthermore, by integrating the microlaboratory with a module for strand displacement amplification (SDA), the identification of the Shiga-like toxin gene (SLT1) from E. coli was accomplished within 2.5 h starting from a dielectrophoretic concentration of intact E. coli bacteria and finishing with an electric-field-driven DNA hybridization assay, detected by fluorescently labeled DNA reporter probes. The integrated microlaboratory can be potentially used in a wide range of applications including detection of bacteria and biowarfare agents, and genetic identification.     

An european pupil project linked to the scientific aims of the experiment AQUARIUS-XENOPUS on the taxi Soyuz flight Andromede to ISS

The German-French biological experiment AQUARIUS-XENOPUS which flew on the Soyuz flight Andromede to the International Space Station ISS (launched October 21, 2001 in Baikonour/Kazakhstan) was extended by an outreach project. Pupils of class 10 to 12 from Ulm/D and Nancy-Tomblaine/F studied swimming behavior of Xenopus tadpoles on ground. They were instructed to perform all experimental steps following the protocol of similar video recordings on ISS. After the flight, they evaluated the kinetics of swimming of both ground controls and space animals. The pupil project included theoretical components to introduce them to the field of gravitational biology. One feature of the project was the exchange of ideas between pupils by meetings which took place in Ulm (June 2001), Nancy (February 2002) and Paris (May 2002). We consider our approach as a successful way to include young people in space experiments on a cheap cost level and to bring ideas of gravitational biology into the curricula of European schools.     

Analysis of simple sequence repeats (SSRs) in wild barley from the Fertile Crescent: associations with ecology,geography and flowering time

Wild barley, Hordeum spontaneum C. Koch, is the progenitor of cultivated barley, Hordeum vulgare. The centre of diversity is in the Fertile Crescent of the Near East, where wild barley grows in a wide range of conditions (temperature, water availability, day length, etc.). The genetic diversity of 39 wild barley genotypes collected from Israel, Turkey and Iran was studied with 33 SSRs of known map location. Analysis of molecular variance (AMOVA) was performed to partition the genetic variation present within from the variation between the three countries of origin. Using classification tree analysis, two (or three) specific SSRs were identified which could correctly classify most of the wild barley genotypes according to country of origin. Associations of SSR variation with flowering time and adaptation to site-of-origin ecology and geography were investigated by two contrasting statistical approaches, linear regression based on SSR length variation and linear regression based on SSR allele class differences. A number of SSRs were significantly associated with flowering time under four different growing regimes (short days, long days, unvernalised and vernalised). Most of the associations observed could be accounted for by close linkage of the SSR loci to earliness per se genes. No associations were found with photoperiodic and vernalisation response genes known to control flowering in cultivated barley suggesting that different genetic factors may be active in wild barley. Novel genomic regions controlling flowering time in wild barley were detected on chromosomes 1HS, 2HL, 3HS and 4HS. Associations of SSRs with site-of-origin ecological and geographic data were found primarily in genomic regions determining plant development. This study shows that the analyses of SSR variation by allele class and repeat length are complementary, and that some SSRs are not necessarily selectively neutral.     

Formation of methylantimony species by an aerobic prokaryote: <Emphasis Type="Italic">Flavobacterium</Emphasis> sp.

Pure cultures of an aerobically grown Flavobacterium sp. were shown by hydride generation-cold trap-atomic absorption spectrometry to biomethylate inorganic antimony (III) supplied as potassium antimony tartrate. Growth inhibition of the Flavobacterium sp. by antimony (III) over the range 0-30 mg Sb l(-1) was assessed by optimising parameters within an extended logistic growth model. Antimony (III) concentrations over this range influenced both the extent of antimony biomethylation (up to 4.0 microg l(-1)) and the relative proportions of the involatile mono-, di, and trimethylantimony species formed. Provision of inorganic arsenic (III) alongside antimony (III) enhanced formation of the involatile methylantimony species up to eight-fold. The data are consistent with accumulation of involatile intermediates from an antimony or arsenic biomethylation pathway in culture supernatants. Low yields of methylantimony species (<0.03%) suggest that antimony biomethylation by the Flavobacterium sp. was a fortuitous rather than a primary resistance mechanism for this element. These findings demonstrate that anaerobiosis is not an obligate requirement for methylantimony formation in prokaryotes, thus broadening the range of habitats for potential formation of methylantimony species in nature.     

The treatment of gastroparesis in the age of the gastric pacemaker: a review

Gastroparesis is a chronic disabling condition of impaired gastric motility that results in decreased quality of life. Currently available medical therapy consists of prokinetic medication combined with antiemetic therapy, dietary modifications, and nutritional supplementation. Many patients continue to have a suboptimal clinical response despite maximal use of these modalities. Instead of surgery, which involves irreversible stomach-modifying procedures, gastric electrical stimulation (GES) with a high-frequency/low-energy stimulus was approved by the US Food and Drug Administration (FDA) and can now be used in this setting. This approach has been shown to decrease symptom frequency and severity, reduce hospitalizations and medical costs, and improve quality of life. Occurrence of complications with this device are uncommon (< 5% of patients). Preliminary studies of new gastric stimulators that restore gastric contractility are promising, but additional investigation is needed. This article reviews the pathophysiology and epidemiology of gastroparesis and the role of conventional medical therapies, and discusses GES therapy with respect to its mechanisms of action, appropriate application, results and benefits, and future directions.     

Hypoxic vasoconstriction of cyclostome systemic vessels: the antecedent of hypoxic pulmonary vasoconstriction?

Hypoxic vasoconstriction (HV) is an intrinsic response of mammalian pulmonary vascular smooth muscle (VSM). In the present study, HV was examined by myography of vessel rings from three primitive vertebrates: New Zealand hagfish (NZH), Pacific hagfish (PH), and sea lamprey (SL). Hypoxia dilated pre-gill arteries (ventral aorta, afferent branchial) from all species, whereas it contracted systemic arteries [dorsal aorta (DA), efferent branchial, celiacomesenteric]. DA HV was reproducible over several days, and it could be sustained in NZH for 8 h without adverse effects. Tension was proportional to PO(2), and half-maximal HV was obtained at PO(2) (mmHg) of 4.7 +/- 0. 2 (NZH), 0.8 +/- 0.1 (PH), and 10.7 +/- 1.9 (SL). HV did not require preconditioning (preexisting contractile stimulus) and was unaffected by elevated extracellular potassium (200 mM NZH; 80 mM SL); removal of the endothelium (NZH); or inhibitors of cyclooxygenase, lipoxygenase, cytochrome P-450 or antagonists of alpha-adrenergic, muscarinic, nicotinic, purinergic, or serotoninergic receptors. These results show that HV is an intrinsic feature of systemic VSM in cyclostomes and suggest that HV has been in the repertoire of VSM responses, since the origin of vertebrates. The exceptionally hardy HV in cyclostome DA may provide a useful model with which to examine both the phylogeny and mechanisms of this response.