Modifications of a conserved regulatory network involving INDEHISCENT controls multiple aspects of reproductive tissue development in Arabidopsis.

Disrupting pollen tube growth and fertilization in Arabidopsis plants leads to reduced seed set and silique size, providing a powerful genetic system with which to identify genes with important roles in plant fertility. A transgenic Arabidopsis line with reduced pollen tube growth, seed set and silique growth was used as the progenitor in a genetic screen to isolate suppressors with increased seed set and silique size. This screen generated a new allele of INDEHISCENT (IND), a gene originally identified by its role in valve margin development and silique dehiscence (pod shatter). IND forms part of a regulatory network that involves several other transcriptional regulators and involves the plant hormones GA and auxin. Using GA and auxin mutants that alter various aspects of reproductive development, we have identified novel roles for IND, its paralogue HECATE3, and the MADS box proteins SHATTERPROOF1/2 in flower and fruit development. These results suggest that modified forms of the regulatory network originally described for the Arabidopsis valve margin, which include these genes and/or their recently evolved paralogs, function in multiple components of GA/auxin-regulated reproductive development.

Hormonal changes during non-climacteric ripening in strawberry.

In contrast to climacteric fruits, where ethylene is known to be pivotal, the regulation of ripening in non-climacteric fruits is not well understood. In the non-climacteric strawberry (Fragaria anannassa), auxin and abscisic acid (ABA) are thought to be important, but the roles of other hormones suggested to be involved in fruit development and ripening are not clear. Here changes in the levels of indole-3-acetic acid (IAA), ABA, GA1, and castasterone from anthesis to fully ripened fruit are reported. The levels of IAA and GA1 rise early in fruit development before dropping to low levels prior to colour accumulation. Castasterone levels are highest at anthesis and drop to very low levels well before ripening commences, suggesting that brassinosteroids do not play an important role in ripening in strawberry. ABA levels are low at anthesis and gradually rise through development and ripening. The synthetic auxin, 1-naphthaleneacetic acid (NAA), can delay ripening, but the application of GA3, the gibberellin biosythesis inhibitor paclobutrazol, and ABA had no significant effect. IAA and ABA levels are higher in the developing achenes than in the receptacle tissue and may be important for receptacle enlargement and ripening, and seed maturation, respectively. Contrary to a recent report, the biologically active GA4 was not detected. The pattern of changes in the levels of the hormones are different from those reported in another well studied non-climateric fruit, grape, suggesting that a single consistent pattern of hormone changes does not occur in this group of fruit during ripening.

The effect of orbital decompression surgery on refraction and intraocular pressure in patients with thyroid orbitopathy.

PURPOSE: To investigate the effect of orbital decompression surgery in thyroid orbitopathy (TO) on both refractive status and intraocular pressure (IOP). PATIENTS AND METHODS: A prospective, multicentre, consecutive audit of patients undergoing thyroid decompression surgery. Indications for surgery included cosmetically unacceptable proptosis or corneal exposure. Exclusion criteria included the following: previous orbital surgery, glaucoma, corneal disease, steroid use in the preceding 12 months, or an acute optic neuropathy. Automated refraction, keratometry, pachymetry, Hertel exophthalmometry, and IOP were recorded at 1 month pre- and 3 months postoperatively. IOP using the Tono-Pen (mean of three readings) was measured in the primary, upgaze, and downgaze positions. RESULTS: Data were collected from 52 orbits of 33 patients (East Grinstead, New York, and Adelaide). There was no significant difference between pre- and postoperative data for sphere, cylinder, or central corneal thickness (CCT). The mean spherical equivalent was -0.43 ± 1.49 D pre-operatively and -0.28 ± 1.52 D postoperatively. The steepest meridian of corneal curvature was 93.1 degrees pre- and 94.2 degrees postoperatively, with no significant difference. Mean IOP significantly decreased when measuring by Goldmann applanation tonometry (GAT) (2.28 mm Hg, (*) P=0.001) and Tono-Pen (3.06 mm Hg, (*) P=<0.0001). IOP measured in upgaze was significantly greater than that in the primary position. Regression analysis between change in IOP and either Hertel exophthalmometry or the number of orbital walls decompressed was non-significant ((*)Student's t-test). CONCLUSION: Patients with TO undergoing orbital decompression had, on average, with-the-rule astigmatism not affected by orbital decompression surgery. IOP was significantly reduced by decompression surgery although no relationship between IOP and the degree of decompression was observed.

Validation of the New Zealand Physical Activity Questionnaire (NZPAQ-LF) and the International Physical Activity Questionnaire (IPAQ-LF) with accelerometry.

BACKGROUND: Validation of instruments used to measure physical activity patterns is essential when attempting to assess the effectiveness of physical activity interventions. OBJECTIVES: To assess the validity of two self-report physical activity questionnaires on a representative sample of New Zealand adults. METHODS: 70 adults aged 18-65 years from around Christchurch, New Zealand were required to wear an ActiGraph GT1M accelerometer during all waking hours for 7 consecutive days. Immediately following the 7 day accelerometer period participants were required to complete the long forms of both the New Zealand Physical Activity Questionnaire (NZPAQ-LF) and the International Physical Activity Questionnaire (IPAQ-LF). RESULTS: Both the NZPAQ-LF and the IPAQ-LF questionnaires showed small to moderate correlations with ActiGraph data for time spent in moderate-intensity physical activity (r=0.19-0.30) and total physical activity (sum of moderate and vigorous-intensity physical activity, r=0.30-0.32). In comparison with the ActiGraph data, both self-report questionnaires tended to overestimate activity levels by approximately 165%. Total physical activity levels gathered from both questionnaires were strongly correlated with each other (r=0.79) and showed good levels of agreement in the Bland-Altman plots. CONCLUSIONS: The long forms of the NZPAQ and IPAQ were found to have acceptable validity when detecting participants' ability to meet activity guidelines based on exercise duration, but a significant amount of overestimation was evident. This presents a need for both instruments to be further developed and tested in order to increase validity.

Secular trends and distributional changes in health and fitness performance variables of 10-14-year-old children in New Zealand between 1991 and 2003.

BACKGROUND: New Zealand children's health and fitness performance is declining over time, but whether this change is because of deterioration in all children's health and fitness performance or can be attributed to just a certain portion of the population, is unknown. OBJECTIVES: In this study, secular trends and distributional changes in health-related and performance-related fitness components among New Zealand primary school children aged 10 to 14 years between 1991 and 2003 were tracked. METHODS: Health- and performance-related fitness parameters including height, weight, body mass index (BMI), flexibility, standing broad jump, 4 x 9-m agility run, abdominal curl-ups, and 550-m run were collected up to twice a year from 3306 children (10-14 years old) from a New Zealand school between 1991 and 2003. RESULTS: Over the 12-year period, the boys' weight increased by 4.5 kg (95% CL 2.7 to 6.2, or 0.8% per year) and girls' by 3.9 kg (95% CL 2.0 to 5.9, or 0.7% per year). Mean BMI increased by 0.12 kg m(-2) (0.6%) and 0.11 kg m(-2) (0.5%) per year for boys and girls, respectively. Children's 550-m run performance declined by 1.5% and 1.7% per year for boys and girls, respectively. Little difference existed between children located in the highest performing and leanest percentiles in 1991 and 2003, but for children in the poorest performing and fattest percentiles, their results were substantially worse in 2003. CONCLUSIONS: These results suggest that the deterioration in the health-related and performance-related fitness components of New Zealand 10-14-year-olds is not homogeneous but skewed towards those children who are the heaviest and perform worst in fitness tests. Previous research on health-related fitness parameters among children in New Zealand is limited but shows secular trends of increasing body mass1 2 in conjunction with deteriorating aerobic fitness performance, muscular endurance and explosive muscular power.3 Internationally, similar increases in body mass have been observed in children since the 1980s.1 4 5 Secular trends of deteriorating health-related fitness performance have also been reported among children around the world,1 5 6 with the most significant decreases observed in aerobic performance. However, trends in health-related variables reported as changes in mean body mass index (BMI) and mean aerobic fitness performance do not reveal possible changes in the distribution of BMI or aerobic performance within the population. Changes in such measures may come about because of a shift in the entire population under investigation or a change in a portion of the population. It is not clear whether New Zealand's entire childhood population is becoming heavier and less aerobically fit or whether only a portion of the children are becoming even heavier and more unfit, with the remaining children showing little secular change. The aim of this study was to track secular trends and distributional changes in body weight and physical fitness parameters among New Zealand primary school children aged 10 to 14 years.

Reducing catheter infections through use of the CD-1000: a retrospective review of a unique catheter specific composite dressing.

PURPOSE: Catheter-related blood stream infections pose a significant risk for patients living with vascular catheters. The cost to manage these infections is substantial. Although the etiology of these infections is multifactorial, tap water has been implicated as a significant causative factor. This retrospective review evaluates the effectiveness of a surgical dressing, the CD-1000, at protecting catheters and exit site wounds from fluid and debris when patients engage in high risk activities like showering. METHODS: All patients who received the CD-1000 from a single national medical supplier from September 2006 through to March 2007 were contacted to participate in this retrospective review; 209 patients, representing 34 states and 175 unique physicians, participated in this study. Effectiveness of the dressing along with prior and current history of catheter events was queried. RESULTS: The CD-1000 was 95% effective at keeping the catheter and exit site dry while patients engaged in high risk activities like showering. Prior to using the CD-1000, the 209 patients reported a historical catheter infection rate of 1.83 per 1000 catheter days. While using the CD-1000 the 209 patients reported a catheter infection rate of 0.47 per 1000 catheter days. CONCLUSION: The CD-1000 catheter specific composite dressing adequately protects vascular catheters and exit sites when patients engage in high risk activities like showering. In this geographically diverse retrospective review, use of the CD-1000 was associated with a 75% reduction in catheter associated infections.

Role of SNAREs and H+-ATPase in the targeting of proton pump-coated vesicles to collecting duct cell apical membrane.

Recycling of H(+)-ATPase to the apical plasma membrane, mediated by vesicular exocytosis and endocytosis, is an important mechanism for controlling H(+) secretion by the collecting duct. We hypothesized that SNAREs (soluble N-ethylmaleimide-sensitive factor attachment proteins) may be involved in the targeting of H(+)-ATPase-coated vesicles. Using a tissue culture model of collecting duct H(+) secretory cells (inner medullary collecting duct (IMCD) cells), we demonstrated that they express the proteins required for SNARE-mediated exocytosis and form SNARE-fusion complexes upon stimulation of H(+)-ATPase exocytosis. Furthermore, exocytic amplification of apical H(+)-ATPase is sensitive to clostridial toxins that cleave SNAREs and thereby inhibit secretion. Thus, SNAREs are critical for H(+)-ATPase cycling to the plasma membrane. The process in IMCD cells has a feature distinct from that of neuronal cells: the SNARE complex includes and requires the vesicular cargo (H(+)-ATPase) for targeting. Using chimeras and truncations of syntaxin 1, we demonstrated that there is a specific cassette within the syntaxin 1 H3 domain that mediates binding of the SNAREs and a second distinct H3 region that binds H(+)-ATPase. Utilizing point mutations of the B1 subunit of the H(+)-ATPase, we document that this subunit contains specific targeting information for the H(+)-ATPase itself. In addition, we found that Munc-18-2, a regulator of exocytosis, plays a multifunctional role in this system: it regulates SNARE complex formation and the affinity of syntaxin 1 for H(+)-ATPase.

Transoesophageal echocardiography for the detection and quantification of pleural fluid in cardiac surgical patients.

BACKGROUND: Transoesophageal echocardiography (TOE) can image pleural fluid. Left pleural collections may be easier to detect than right, as the thoracic aorta serves as an acoustic window. Attempts to quantify pleural fluid using TOE are restricted to a case report in which volume was predicted by multiplying maximal cross-sectional area (CSA(max)) by axial length (AL). A computed tomography (CT) derived formula for quantifying pleural effusions is maximal effusion depth squared (d2) multiplied by maximal effusion length. METHODS: Eight patients were studied before chest closure following coronary bypass surgery. Fifty millilitre saline aliquots were instilled into the pleural space until detected by TOE. Saline was then instilled up to the next 200 ml increment and further 200 ml aliquots added until it spilled from the pleural space. CSA(max), d and AL were measured for each stage and used to calculate pleural fluid volume. RESULTS: Median detection volume (range) was 125 ml (50-200) on the left and 225 ml (150-300) on the right (P = 0.016). Volume calculated by CSA(max) x AL correlated strongly with actual volume (r2 = 0.93 left and 0.92 right) as did volume calculated by d2 x AL (r2 = 0.86 left and 0.89 right). Mean difference between volume calculated by CSA(max) x AL and actual volume was - 51 ml on the left and 45 ml on the right vs - 253 ml on the left and - 212 ml on the right for volume calculated by d2 x AL. CONCLUSIONS: TOE detects small volumes of pleural fluid on both sides of the chest. CSA(max) x AL provides a reasonably accurate measure of pleural fluid volume.

Hodgkin' s disease with fulminant non-alcoholic steatohepatitis.

We report a woman with daily febrile episodes who developed fulminant hepatic failure. A percutaneous liver biopsy demonstrated non-alcoholic steatohepatitis, with no evidence of neoplastic infiltration. Post-mortem examination revealed stage IV Hodgkin's disease with trivial liver involvement. Rapidly progressive steatohepatitis causing acute liver failure may be a paraneoplastic presentation of Hodgkin's disease, possibly mediated by cytokines.

ECM gene expression correlates with in vitro tissue growth and development in fibrin gel remodeled by neonatal smooth muscle cells.

A tissue growth and development process occurred in neonatal SMC-fibrin gel constructs when cultured in DMEM supplemented with TGF-beta1 and insulin over a 5 week period. These constructs may thus serve as the basis for cardiovascular tissue replacements and future models of cardiovascular tissue growth, repair and regeneration. Following fibrin gel contraction during week 1, peak rates of SMC proliferation, collagen production and tropoelastin production occurred between weeks 1-4. Organized, cross-linked collagen and elastic fibers replaced the degrading fibrin over weeks 3-5 and were manifested as increased mechanical strength. The peak rate of SMC proliferation (weeks 1-2) preceded that for maximum collagen production (weeks 2-4), which was consistent with the 3 week time point of maximum expression of collagen type I and III from qRT-PCR. Insoluble elastin quantification revealed that the majority of elastic fibers were produced by week 4, which was also consistent with the qRT-PCR data showing a dramatic down-regulation of tropoelastin expression by week 4, indicating elastogenesis occurred during the early stages of tissue growth and development. There was a strong up-regulation of lysyl oxidase expression during weeks 1-3 with a peak in expression at week 3, correlating with the phases of collagen and tropoelastin production. An increase in MMP-2 expression over weeks 1-5 suggested an increase in ECM remodeling as the tissue developed. Mechanical strength doubled over weeks 4-5 when production of collagen and elastic fibers and expression of lysyl oxidase were subsiding. This may have been due in part to the more organized collagen fibrils evident from the histological sections in weeks 3-5.

Twisted gastrulation is a conserved extracellular BMP antagonist.

Bone morphogenetic protein (BMP) signalling regulates embryonic dorsal-ventral cell fate decisions in flies, frogs and fish. BMP activity is controlled by several secreted factors including the antagonists chordin and short gastrulation (SOG). Here we show that a second secreted protein, Twisted gastrulation (Tsg), enhances the antagonistic activity of Sog/chordin. In Drosophila, visualization of BMP signalling using anti-phospho-Smad staining shows that the tsg and sog loss-of-function phenotypes are very similar. In S2 cells and imaginal discs, TSG and SOG together make a more effective inhibitor of BMP signalling than either of them alone. Blocking Tsg function in zebrafish with morpholino oligonucleotides causes ventralization similar to that produced by chordin mutants. Co-injection of sub-inhibitory levels of morpholines directed against both Tsg and chordin synergistically enhances the penetrance of the ventralized phenotype. We show that Tsgs from different species are functionally equivalent, and conclude that Tsg is a conserved protein that functions with SOG/chordin to antagonize BMP signalling.

Gibberellin biosynthesis mutations and root development in pea.

Dwarf mutants of pea (Pisum sativum), with impaired gibberellin (GA) biosynthesis in the shoot, were studied to determine whether the roots of these genotypes had altered elongation and GA levels. Mutations na, lh-2, and ls-1 reduced GA levels in root tips and taproot elongation, although in lh-2 and ls-1 roots the reduction in elongation was small (less than 15%). The na mutation reduced taproot length by about 50%. The roots of na plants elongated in response to applied GA(1) and recombining na with mutation sln (which blocks GA catabolism) increased GA(1) levels in root tips and completely restored normal root development. In shoots, Mendel's le-1 mutation impairs the 3beta-hydroxylation of GA(20) to the bioactive GA(1), resulting in dwarfism. However, GA(1) and GA(20) levels were normal in le-1 roots, as was root development. The null mutation le-2 also did not reduce root GA levels or elongation. The results support the theory that GAs are important for normal root elongation in pea, and indicate that a 3beta-hydroxylase gene other than LE operates in pea roots.

Fluid balance during and after an ironman triathlon.

OBJECTIVE: To record weight changes, fluid intake and changes in serum sodium concentration in ultradistance triathletes. DESIGN: Descriptive research. SETTING: Ironman triathlon (3.8 km swim, 180 km cycle, 42.2 km run). Air temperature at 1200 h was 21 degrees C, (relative humidity 91%). Water temperature was 20.7 degrees C. PARTICIPANTS: 18 triathletes. INTERVENTIONS: None. MAIN OUTCOME MEASURES: Subjects were weighed and had blood drawn for serum sodium concentration [Na], hemoglobin, and hematocrit, pre-race, post-race, and at 0800 h on the morning following the race ("recovery"); subjects were also weighed at transitions. Fluid intake during the race was estimated by athlete recall. RESULTS: Median weight change during the race = -2.5 kg (p < 0.0006). Subjects lost weight during recovery (median = -1.0 kg) (p < 0.03). Median hourly fluid intake = 716 ml/h (range 421-970). Fluid intakes were higher on the bike than on the run (median 889 versus 632 ml/h, p = 0.03). Median calculated fluid losses cycling were 808 ml/h and running were 1,021 ml/h. No significant difference existed between pre-race and post-race [Na] (median 140 versus 138 mmol/L) or between post-race and recovery [Na] (median 138 versus 137 mmol/L). Plasma volume increased during the race, median + 10.8% (p = 0.0005). There was an inverse relationship between change in [Na] pre-race to post-race and relative weight change (r = -0.68, p = 0.0029). Five subjects developed hyponatremia ([Na] 128-133 mmol/L). CONCLUSIONS: Athletes lose 2.5 kg of weight during an ultradistance triathlon. most likely from sources other than fluid loss. Fluid intakes during this event are more modest than that recommended for shorter duration exercise. Plasma volume increases during the ultradistance triathlon. Subjects who developed hyponatremia had evidence of fluid overload despite modest fluid intakes.

Auxin promotes gibberellin biosynthesis in decapitated tobacco plants.

Excision of the apical bud (decapitation) of tobacco (Nicotiana tabacum L.) plants reduced the endogenous levels of indole-3-acetic acid (IAA), gibberellin A20 (GA20), and GA1 (the bioactive GA), in internode tissue below the excision site. Application of IAA to the stump of decapitated plants dramatically increased GA20 content, to a level 3-fold greater than in intact plants. Gibberellin A1 content was also increased by IAA. Decapitation reduced the conversion of [14C]GA19 to [14C]GA20 and of [14C]GA20 to [14C]GA1, and appeared to promote the deactivation pathway [14C]GA20 to [14C]GA29 to [14C]GA29-catabolite. Application of auxin counteracted these effects, but did not restore the conversion of [14C]GA20 to [14C]GA1 to the level found in intact plants. The results indicate that auxin is necessary for normal GA biosynthesis in stems of tobacco.

Cutaneous blastomycosis in New Brunswick: case report.

Blastomycosis is a fungal infection of immunocompetent hosts. We present a case of cutaneous blastomycosis acquired in New Brunswick, which provides evidence that this disease is endemic in Atlantic Canada. This case also demonstrates that the diagnosis of blastomycosis may be elusive. Perseverance, a high index of clinical suspicion and close cooperation with the microbiology laboratory may be required to diagnose this uncommon condition.

Changes in gibberellin A(1) levels and response during de-etiolation of pea seedlings.

The level of gibberellin A(1) (GA(1)) in shoots of pea (Pisum sativum) dropped rapidly during the first 24 h of de-etiolation. The level then increased between 1 and 5 d after transfer to white light. Comparison of the metabolism of [(13)C(3)H] GA(20) suggested that the initial drop in GA(1) after transfer is mediated by a light-induced increase in the 2beta-hydroxylation of GA(1) to GA(8). A comparison of the elongation response to GA(1) at early and late stages of de-etiolation provided strong evidence for a change in GA(1) response during de-etiolation, coinciding with the return of GA(1) levels to the normal, homeostatic levels found in light- and dark-grown plants. The emerging picture of the control of shoot elongation by light involves an initial inhibition of elongation by a light-induced decrease in GA(1) levels, with continued inhibition mediated by a light-induced change in the plant's response to the endogenous level of GA(1). Hence the plant uses a change in hormone level to respond to a change in the environment, but over time, homeostasis returns the level of the hormone to normal once the ongoing change in environment is accommodated by a change in the response of the plant to the hormone.

Evidence that auxin promotes gibberellin A1 biosynthesis in pea.

In shoots of the garden pea, the bioactive gibberellin (GA1) is synthesised from GA20, and the enzyme which catalyses this step (a GA 3-oxidase -- PsGA3ox1) is encoded by Mendel's LE gene. It has been reported previously that decapitation of the shoot (excision of the apical bud) dramatically reduces the conversion of [3H]GA20 to [3H]GA1 in stems, and here we show that endogenous GA1 and PsGA3ox1 transcript levels are similarly reduced. We show also that these effects of decapitation are completely reversed by application of the auxin indole-3-acetic acid (IAA) to the 'stump' of decapitated plants. Gibberellin A20 is also converted to an inactive product, GA29, and this step is catalysed by a GA 2-oxidase, PsGA2ox1. In contrast to PsGA3ox1, PsGA2ox1 transcript levels were increased by decapitation and reduced by IAA application. Decapitation and IAA treatment did not markedly affect the level of GA1 precursors. It is suggested that in intact pea plants, auxin from the apical bud moves into the elongating internodes where it (directly or indirectly) maintains PsGA3ox1 transcript levels and, consequently, GA1 biosynthesis.