Distribution of mitochondria within Muller cells – II. Post-natal development of the rabbit retinal periphery in vivo and in vitro: dependence on oxygen supply

The occurrence and localization of mitochondria within glial (Muller) cells and neurons of the peripheral (avascular) rabbit retina was studied electron microscopically and by immunocytochemical demonstration of the mitochondrial enzyme GABA transaminase (GABA-T). Post-natal development in vivo was compared with development of organ cultures from neonatal rabbit retinae, grown over 2 weeks in vitro. The adult pattern of mitochondrial localization (restriction to the sclerad end of the cells) was observed from the beginning of enzyme expression at early post-natal stages. However, when neonatal retinal pieces were grown in vitro with their vitread surface exposed to the air, their Muller cells contained mitochondria along most of their length. When functionally developed retinae from postnatal day 14 were explanted in vitro, they retained their sclerad mitochondrial distribution for almost 24 h but thereafter the inner portions of their cytoplasm became occupied by mitochondria within a few hours. This was achieved mainly by mitochondrial migration rather than by formation of new mitochondria because it was not prevented by cycloheximide-induced inhibition of protein synthesis. These data support the following hypotheses: (1) the mitochondrial distribution in Muller cells is determined by the local cytoplasmic O2 pressure (pO2), (2) existing mitochondria move towards cytoplasmic regions of sufficient pO2 by rather rapid migration and (3) the start of this migration is delayed by almost 24 h due to the action of as yet unknown control mechanisms. In contrast, the mitochondrial content of retinal ganglion and amacrine cells in the vitread retinal layers was virtually independent of the source and level of oxygen supply.     

The Arabidopsis transposable element Tag1 is widely distributed among Arabidopsis ecotypes

Tag1 is an autonomous transposable element (3.3 kb in length) first identified as an insertion in the CHL1 (NRT1) gene of Arabidopsis thaliana. Tag1 has been found in the Landsberg erecta ecotype of A. thaliana but not in Columbia or WS. In this paper, 41 additional ecotypes were examined for the presence of Tag1. Using an internal Tag1 fragment as probe, we found that DNA from 19 of the 41 ecotypes strongly hybridized to Tag1. Almost all of the Tag1-containing ecotypes had only one or two copies of Tag1 per haploid genome, as determined by Southern blot analysis. The only exception, Bf-1 from Bretagny-sur-Orge, France, had four copies. Two ecotypes, Di-G and S96, gave identical Southern blot patterns to that of Landsberg erecta and were subsequently shown to contain Tag1 at the same two positions found in Landsberg erecta (loci designated as Tag1-2 and Tag1-3). Two other ecotypes, Ag-0 and Lo-1, had a Tag1 element located at Tag1-2 but not at Tag1-3. The distance between these two loci was determined to be 0.37 cM. Analysis of DNA from two related species, A. griffithiana and A. pumila, showed that both species contain sequences that hybridize to Tag1 and that could be amplified with an oligonucleotide specific to the terminal inverted repeats of Tag1. These results show that Tag1 and related elements are present, and may be useful for insertional mutagenesis, in many A. thaliana ecotypes and several Arabidopsis species.     

Albumin synthesis after intense intermittent exercise in human subjects

Yang, Roger C., Gary W. Mack, Robert R. Wolfe, and Ethan R. Nadel. Albumin synthesis after intense intermittent exercise inhuman subjects. J. Appl. Physiol.84(2): 584-592, 1998.We measured hepatic albumin synthesis infive volunteers (4 men and 1 woman) at 3 and 6 h after recovery fromintense exercise. A primed-constant infusion of a stable isotopictracer of phenylalanine was used to determine hepatic fractionalsynthetic rate (FSR) and absolute synthetic rate (ASR) of albumin fromthe enrichment of phenylalanine in albumin. The infusion of the stableisotope tracer began 2 h after upright exercise or upright rest.Albumin FSR and ASR were 6.39 ± 0.48%/day and 120 ± 9 mg · kg bodywt¹ · day¹,respectively, 3-6 h after recovery from exercise; the FSR and ASRon the time control study day were 5.94 ± 0.47%/day and 104 ± 9 mg · kg bodywt¹ · day¹,respectively. The 6 and 16% increases(P < 0.05) in FSR and ASR afterexercise were associated with an elevated plasma albumin content at 5 and 6 h of recovery (P < 0.05), anincreased total protein content throughout recovery(P < 0.05), and a negative freewater clearance (P < 0.05) at 2, 3, and 6.5 h of recovery compared with baseline values; these variableswere unchanged from their baselines on the time control study day.Increased albumin content and reduced free water clearance contributeto a retention of fluid within the circulation after intense exercise. The measured increase in albumin synthesis could not account for theentire increase in albumin content at 6 h of recovery from exercise.However, we estimate that if the increased activity was maintained forthe next 18 h, it could account for the expected increase in albumincontent at 24 h of recovery.

     

A comparison of techniques for isolation of the outer membrane proteins of Haemophilus influenzae type b

We compared several rapid techniques used for extraction of outer membrane proteins from gram-negative enteric bacteria to Haemophilus influenzae type b. After lysis of cells with a French press, the inner and outer membranes were separated by isopycnic centrifugation. Each membrane was identified by density, morphology, enzymatic activity, and susceptibility to solid-phase iodination of intact cells. By sodium dodecyl sulfate-polyacrylamide gel electrophoresis, we identified 10 polypeptides which were enriched in the outer membrane band compared to the inner membrane band. Using these proteins, we compared the polypeptide pattern of outer membranes with that obtained by (1) selective solubilization with sodium dodecyl-beta-D-maltoside, octyl-beta-D-glucopyranoside, Triton X-100, sodium, or cholamidopropyl dimethylaminopropanesulfonate; (2) extraction with chaotropic agents and heat; and (3) differential centrifugation of vesicles shed during transition from log growth phase to stationary growth phase. There were definable differences between the polypeptide pattern of membranes obtained with each rapid technique compared to the polypeptide pattern of isolated outer membranes. The polypeptide pattern of lithium extracts and the Triton X-100 insoluble fractions of total membranes most closely approximated the polypeptide pattern of isopycnically isolated outer membranes. Depending on the outer membrane protein sought, one of these rapid techniques can be utilized when a rapid method of outer membrane protein isolation is required.     

Did fleshy fruit pulp evolve as a defence against seed loss rather than as a dispersal mechanism?

Relatively few studies have examined the evolution of the mutualism between endozoochorous plants and seed dispersers. Most seed dispersal studies are ecological and examine the role of fruit pulp in promoting seed dispersal. This interaction is often assumed to have originated due to selection stemming from seed dispersers. Here I suggest a "defence scenario" wherein fleshy fruits originated as mechanisms to defend seeds and secondarily became structures to promote seed dispersal. I suggest that frugivory followed from herbivores that specialized on consuming seed defensive tissues and that enhanced seed dispersal was initially a consequence of seed defence. The proposed defence scenario is not posited as an explanation for the sequence that led to all modern frugivores. However, it is suggested that seed predation was the initial source of selection that led to fleshy fruits; the necessary precursor to frugivory. Support is described from the fossil record and from modern structures and interactions. Testable predictions are made in hope that greater interest will be focused on the defensive role of fleshy fruit pulp both in modern interactions and historically.     

Representations of Nature on the Mongolian Steppe: An Investigation of Scientific Knowledge Construction

Extensive land degradation across the Mongolian steppe has prompted a variety of multinational and multidisciplinary research projects over recent years. The situation provides an important opportunity to investigate and illuminate some of the international, national, and local dimensions of scientific practice that critically condition the production of environmental knowledge. In this article I juxtapose the competing knowledge bases and assumptions of various relevant groups (including natural vs. social scientists, nationalist Chinese vs. ethnic Mongolians, and urban intellectuals vs. indigenous herders) to develop the argument that multiple ideological and institutional boundaries work together to circumscribe scientific inquiry and data collection. The situated construction of knowledge undermines prospects for improving incremental objectivity and impedes more comprehensive understanding of serious environmental problems. [Chinese grasslands, land degradation, indigenous knowledge, sociology of science, interdisciplinary research]     

Association between genome-wide association studies reported SNPs and pediatric-onset Crohn’s disease in Canadian children

A recent pediatric-focused genome-wide association study has implicated three novel susceptibility loci for Crohn’ disease (CD).We aimed to investigate whether the three recently reported and other previously reported genes/loci were also associated with CD in Canadian children. A case–control design was implemented at three pediatric gastroenterology clinics in Canada. Children <19 years of age with a confirmed diagnosis of CD were recruited along with controls. Single nucleotide polymorphisms (SNPs) in 19 reported genes/loci were genotyped. Associations between individual SNPs and CD were examined. A total of 563 cases and 553 controls were studied. The mean (±SD) age of the cases was 12.3 (±3.2) years. Most cases were male (56.0%), had ileo-colonic disease (L3 ± L4, 48.8%) and inflammatory behavior (B1 ± p, 87.9%) at diagnosis. Allelic association analysis (two-tailed) showed that 8 of the 19 targeted SNPs were significantly associated with overall susceptibility for CD. Associations with one additional SNP was borderline non-significant. Significantly associated SNPs included SNPs rs1250550 (p = 0.026) and rs8049439 (p = 0.04), recently reported to be specifically associated with pediatric-onset CD.Based on the results, we confirmed associations between two of the three novel pediatric-CD loci and other regions reported for associations with either pediatric and/or adult-onset CD.     

Transcapillary escape rate of albumin in humans during exercise-induced hypervolemia

Haskell, Andrew, Ethan R. Nadel, Nina S. Stachenfeld, KeiNagashima, and Gary W. Mack. Transcapillary escape rate of albuminin humans during exercise-induced hypervolemia. J. Appl. Physiol. 83(2): 407-413, 1997.To test thehypotheses that plasma volume (PV) expansion 24 h after intenseexercise is associated with reduced transcapillary escape rate ofalbumin (TER_alb) and that localchanges in transcapillary forces in the previously active tissues favorretention of protein in the vascular space, we measured PV,TER_alb, plasma colloid osmoticpressure (COP_p), interstitialfluid hydrostatic pressure (Pi), and colloid osmotic pressure in legmuscle and skin and capillary filtration coefficient (CFC) in the armand leg in seven men and women before and 24 h after intense uprightcycle ergometer exercise. Exercise expanded PV by 6.4% at 24 h (43.9 ± 0.8 to 46.8 ± 1.2 ml/kg, P < 0.05) and decreased total protein concentration (6.5 ± 0.1 to6.3 ± 0.1 g/dl, P < 0.05) andCOP_p (26.1 ± 0.8 to 24.3 ± 0.9 mmHg, P < 0.05), although plasmaalbumin concentration was unchanged. TER_alb tended to decline (8.4 ± 0.5 to 6.5 ± 0.7%/h, P = 0.11) and was correlated with the increase in PV(r = 0.69,P < 0.05). CFC increased in the leg(3.2 ± 0.2 to 4.3 ± 0.5 µl · 100 g¹ · min¹ · mmHg¹,P < 0.05), and Pi showed a trend toincrease in the leg muscle (2.8 ± 0.7 to 3.8 ± 0.3 mmHg, P = 0.08). These datademonstrate that TER_alb isassociated with PV regulation and that local transcapillary forcesin the leg muscle may favor retention of albumin in the vascular spaceafter exercise.

     

Novel IL10 gene family associations with systemic juvenile idiopathic arthritis

Juvenile idiopathic arthritis (JIA) is the most common cause of chronic childhood disability and encompasses a number of disease subgroups. In this study we have focused on systemic JIA (sJIA), which accounts for approximately 11% of UK JIA cases. This study reports the investigation of three members of the IL10 gene family as candidate susceptibility loci in children with sJIA. DNA from 473 unaffected controls and 172 patients with sJIA was genotyped for a single nucleotide polymorphism (SNP) in IL19 and IL20 and two SNPs in IL10. We examined evidence for association of the four SNPs by single marker and haplotype analysis. Significant differences in allele frequency were observed between cases and controls, for both IL10-1082 (p = 0.031) and IL20-468 (p = 0.028). Furthermore, examination of the haplotypes of IL10-1082 and IL20-468 revealed greater evidence for association (global p = 0.0006). This study demonstrates a significant increased prevalence of the low expressing IL10-1082 genotype in patients with sJIA. In addition, we show a separate association with an IL20 polymorphism, and the IL10-1082A/IL20-468T haplotype. The two marker 'A-T' haplotype confers an odds ratio of 2.24 for sJIA. This positive association suggests an important role for these cytokines in sJIA pathogenesis.     

Autoantibodies to components of the mitotic apparatus

Autoantibodies directed to a variety of cellular antigens and organelles are a feature of autoimmune diseases. They have proven useful in a clinical setting to establish diagnosis, estimate prognosis, follow disease progression, alter therapy, and initiate new investigations. Cellular and molecular biologists have used autoantibodies as probes to identify molecules involved in key cellular processes. One of the most interesting sets of autoantibodies are those that target antigens within the mitotic apparatus (MA). The MA includes chromosomes, spindle microtubules and centrosomes. The identification, localization, function, and clinical relevance of MA autoantigens is the focus of this review. Abbreviations: ATP – adenosine triphosphate; CENP – centromere protein; CREST – calcinosis, Raynaud's phenomenon, esophageal dysmotility, sclerodactyly, telangiectasia; HMG – high mobility group; IB – intercellular bridge; IIF – indirect immunofluorescence; MAPs – microtubule associated proteins; NuMA – nuclear mitotic apparatus; NOR – nucleolar organizer; PBC – primary biliary cirrhosis; PM – polymyositis; Pol I, II, III – RNA polymerases; RA-rheumatoid arthritis; SLE – systemic lupus erythematosus; SS – Sjögren's syndrome; SSc – systemic sclerosis; topo – topoisomerase.