The effect of high-flow nasal oxygen flow rate on gas exchange in apnoeic patients: a randomised controlled trial.

High-flow nasal oxygen can be administered at induction of anaesthesia for the purposes of pre-oxygenation and apnoeic oxygenation. This intervention is claimed to enhance carbon dioxide elimination during apnoea, but the extent to which this occurs remains poorly quantified. The optimal nasal oxygen flow rate for gas exchange is also unknown. In this study, 114 patients received pre-oxygenation with high-flow nasal oxygen at 50 l.min-1. At the onset of apnoea, patients were allocated randomly to receive one of three nasal oxygen flow rates: 0 l.min-1; 70 l.min-1; or 120 l.min-1. After 4 minutes of apnoea, all oxygen delivery was ceased, tracheal intubation was performed, and oxygen delivery was recommenced when SpO2 was 92%. Mean (SD) PaCO2 rise during the first minute of apnoea was 1.39 (0.39) kPa, 1.41 (0.29) kPa, and 1.26 (0.38) kPa in the 0 l.min-1, 70 l.min-1 and 120 l.min-1 groups, respectively; p = 0.16. During the second, third and fourth minutes of apnoea, mean (SD) rates of rise in PaCO2 were 0.34 (0.08) kPa.min-1, 0.36 (0.06) kPa.min-1 and 0.37 (0.07) kPa.min-1 in the 0 l.min-1, 70 l.min-1 and 120 l.min-1 groups, respectively; p = 0.17. After 4 minutes of apnoea, median (IQR [range]) arterial oxygen partial pressures in the 0 l.min-1, 70 l.min-1 and 120 l.min-1 groups were 24.5 (18.6-31.4 [12.3-48.3]) kPa; 36.6 (28.1-43.8 [9.8-56.9]) kPa; and 37.6 (26.5-45.4 [11.0-56.6]) kPa, respectively; p < 0.001. Median (IQR [range]) times to desaturate to 92% after the onset of apnoea in the 0 l.min-1, 70 l.min-1 and 120 l.min-1 groups, were 412 (347-509 [190-796]) s; 533 (467-641 [192-958]) s; and 531 (462-681 [326-1007]) s, respectively; p < 0.001. In conclusion, the rate of carbon dioxide accumulation in arterial blood did not differ significantly between apnoeic patients who received high-flow nasal oxygen and those who did not.

Serum OPG/TRAIL ratio predicts the presence of cardiovascular disease in people with type 2 diabetes mellitus.

AIMS: Cardiovascular disease (CVD) is the leading cause of mortality in type 2 diabetes mellitus (T2DM). Epidemiological studies suggest serum Osteoprotegrin (OPG)/Tumour-necrosis-factor-related-apoptosis-inducing- ligand (TRAIL) ratio may be a useful marker of cardiovascular risk. This study aimed to compare serum levels of TRAIL, OPG and OPG/TRAIL ratio in people with T2DM, with and without a history of CVD, and controls; and to determine which of these indices, if any, predict cardiovascular risk. METHODS: In this single centre observational study of 133 participants, people with T2DM, with and without a history of a cardiovascular event in the last 5 years, were recruited along with a control cohort without T2DM or CVD. Demographic information and anthropometric measurements were recorded. Blood samples were taken and OPG and TRAIL were measured using ELISA. RESULTS: People with T2DM and CVD had higher OPG/TRAIL ratios compared to controls or those with a new diagnosis of T2DM. After adjustment for potential confounding factors, OPG/TRAIL ratio was significantly associated with the presence of CVD in people with T2DM and an OPG/TRAIL ratio cut-off > 38.6 predicted the presence of CVD in this cohort with a sensitivity of 80% and specificity of 82%. CONCLUSION: This study suggests that OPG/TRAIL ratio may have a role as a biomarker of CVD in people with T2DM.

Pre-oxygenation with facemask oxygen vs high-flow nasal oxygen vs high-flow nasal oxygen plus mouthpiece: a randomised controlled trial.

High-flow nasal oxygen used before and during apnoea prolongs time to desaturation at induction of anaesthesia. It is unclear how much oxygenation before apnoea prolongs this time. We randomly allocated 84 participants to 3 minutes of pre-oxygenation by one of three methods: 15 l.min-1 by facemask; 50 l.min-1 by high-flow nasal cannulae only; or 50 l.min-1 by high-flow nasal cannulae plus 15 l.min-1 by mouthpiece. We then anaesthetised and intubated the trachea of 79 participants and waited for oxygen saturation to fall to 92%. Median (IQR [range]) times to desaturate to 92% after pre-oxygenation with facemask oxygen, high-flow nasal oxygen only and high-flow nasal oxygen with mouthpiece, were: 309 (208-417 [107-544]) s; 344 (250-393 [194-585]) s; and 386 (328-498 [182-852]) s, respectively, p = 0.014. Time to desaturation after facemask pre-oxygenation was shorter than after combined nasal and mouthpiece pre-oxygenation, p = 0.006. We could not statistically distinguish high-flow nasal oxygen without mouthpiece from the other two groups for this outcome. Median (IQR [range]) arterial oxygen partial pressure after 3 minutes of pre-oxygenation by facemask, nasal cannulae and nasal cannulae plus mouthpiece, was: 49 (36-61 [24-66]) kPa; 57 (48-62 [30-69]) kPa; and 61 (55-64 [36-72]) kPa, respectively, p = 0.003. Oxygen partial pressure after 3 minutes of pre-oxygenation with nasal and mouthpiece combination was greater than after facemask pre-oxygenation, p = 0.002, and after high-flow nasal oxygen alone, p = 0.016. We did not reject the null hypothesis for the pairwise comparison of facemask pre-oxygenation and high-flow nasal pre-oxygenation, p = 0.14.

Associations between maternal physical activity in early and late pregnancy and offspring birth size: remote federated individual level meta-analysis from eight cohort studies.

OBJECTIVE: Evidence on the impact of leisure time physical activity (LTPA) in pregnancy on birth size is inconsistent. We aimed to examine the association between LTPA during early and late pregnancy and newborn anthropometric outcomes. DESIGN: Individual level meta-analysis, which reduces heterogeneity across studies. SETTING: A consortium of eight population-based studies (seven European and one US) comprising 72 694 participants. METHODS: Generalised linear models with consistent inclusion of confounders (gestational age, sex, parity, maternal age, education, ethnicity, BMI, smoking, and alcohol intake) were used to test associations between self-reported LTPA at either early (8-18 weeks gestation) or late pregnancy (30+ weeks) and the outcomes. Results were pooled using random effects meta-analyses. MAIN OUTCOME MEASURES: Birth weight, large-for-gestational age (LGA), macrosomia, small-for-gestational age (SGA), % body fat, and ponderal index at birth. RESULTS: Late, but not early, gestation maternal moderate to vigorous physical activity (MVPA), vigorous activity, and LTPA energy expenditure were modestly inversely associated with BW, LGA, macrosomia, and ponderal index, without heterogeneity (all: I2  = 0%). For each extra hour/week of MVPA, RR for LGA and macrosomia were 0.97 (95% CI: 0.96, 0.98) and 0.96 (95% CI: 0.94, 0.98), respectively. Associations were only modestly reduced after additional adjustments for maternal BMI and gestational diabetes. No measure of LTPA was associated with risk for SGA. CONCLUSIONS: Physical activity in late, but not early, pregnancy is consistently associated with modestly lower risk of LGA and macrosomia, but not SGA. TWEETABLE ABSTRACT: In an individual participant meta-analysis, late pregnancy moderate to vigorous physical activity modestly reduced birth size outcomes.

Higher rate of fat oxidation during rowing compared with cycling ergometer exercise across a range of exercise intensities.

The relative contribution of carbohydrate and fat oxidation to energy expenditure during exercise is dependent on variables including exercise intensity, mode, and recruited muscle mass. This study investigated patterns of substrate utilization during two non-weightbearing exercise modalities, namely cycling and rowing. Thirteen young, moderately trained males performed a continuous incremental (3-min stages) exercise test to exhaustion on separate occasions on an electronically braked cycle (CYC) ergometer and an air-braked rowing (ROW) ergometer, respectively. On two further occasions, participants performed a 20-min steady-state exercise bout at ∼50%VO2peak on the respective modalities. Despite similar oxygen consumption, rates of fat oxidation (FATox ) were ∼45% higher during ROW compared with CYC (P < 0.05) across a range of power output increments. The crossover point for substrate utilization occurred at a higher relative exercise intensity for ROW than CYC (57.8 ± 2.1 vs 42.1 ± 3.6%VO2peak , P < 0.05). During steady-state submaximal exercise, the higher FATox during ROW compared with CYC was maintained (P < 0.05), but absolute FATox were 42% (CYC) and 28% (ROW) lower than during incremental exercise. FATox is higher during ROW compared with CYC exercise across a range of exercise intensities matched for energy expenditure, and is likely as a consequence of larger muscle mass recruited during ROW.

Development of a DNA Macroarray for the Detection and Identification of Fungal Pathogens Causing Decline of Young Grapevines.

Young vine decline (YVD) is a complex disease caused by at least 51 different fungi and responsible for important economic losses to the grapevine industry worldwide. YVD fungi are known to occur in planting material. Hence, detection prior to planting is critical to assure longevity of newly established vineyards. A DNA macroarray based on reverse dot-blot hybridization containing 102 oligonucleotides complementary to portions of the ß-tubulin region was developed for detection of YVD fungi. Specificity of the array was first evaluated against 138 pure fungal cultures representing 72 different taxa from nine genera, including 37 YVD species. In total, 61 species, including 34 YVD pathogens, were detected and identified by the array. The detection limit of the array was below 0.1 pg of genomic DNA. The array was validated against artificially inoculated canes and soil and commercial planting material, with the latter showing a high incidence of YVD fungi in nursery plants otherwise not detected by traditional plating and culturing. This DNA array proved to be a rapid and specific tool to simultaneously detect and identify most YVD fungi in a single test, which has the potential to be used in commercial diagnostics or by the grapevine nursery industry to determine the health status of the planting material.

Islet oxygen consumption rate dose predicts insulin independence for first clinical islet allotransplants.

BACKGROUND: Human islet allotransplantation for the treatment of type 1 diabetes is in phase III clinical trials in the U.S. and is the standard of care in several other countries. Current islet product release criteria include viability based on cell membrane integrity stains, glucose-stimulated insulin release, and islet equivalent (IE) dose based on counts. However, only a fraction of patients transplanted with islets that meet or exceed these release criteria become insulin independent following 1 transplant. Measurements of islet oxygen consumption rate (OCR) have been reported as highly predictive of transplant outcome in many models. METHOD: In this article we report on the assessment of clinical islet allograft preparations using OCR dose (or viable IE dose) and current product release assays in a series of 13 first transplant recipients. The predictive capability of each assay was examined and successful graft function was defined as 100% insulin independence within 45 days post-transplant. RESULTS: OCR dose was most predictive of CTO. IE dose was also highly predictive, while glucoses stimulated insulin release and membrane integrity stains were not. CONCLUSION: OCR dose can predict CTO with high specificity and sensitivity and is a useful tool for evaluating islet preparations prior to clinical human islet allotransplantation.

Human islet viability and function is maintained during high-density shipment in silicone rubber membrane vessels.

BACKGROUND: The shipment of human islets (IE) from processing centers to distant laboratories is beneficial for both research and clinical applications. The maintenance of islet viability and function in transit is critically important. Gas-permeable silicone rubber membrane (SRM) vessels reduce the risk of hypoxia-induced death or dysfunction during high-density islet culture or shipment. SRM vessels may offer additional advantages: they are cost-effective (fewer flasks, less labor needed), safer (lower contamination risk), and simpler (culture vessel can also be used for shipment). METHOD: IE were isolated from two manufacturing centers and shipped in 10-cm(2) surface area SRM vessels in temperature- and pressure-controlled containers to a distant center after at least 2 days of culture (n = 6). Three conditions were examined: low density (LD), high density (HD), and a microcentrifuge tube negative control (NC). LD was designed to mimic the standard culture density for IE preparations (200 IE/cm(2)), while HD was designed to have a 20-fold higher tissue density, which would enable the culture of an entire human isolation in 1-3 vessels. Upon receipt, islets were assessed for viability (measured by oxygen consumption rate normalized to DNA content [OCR/DNA)]), quantity (measured by DNA), and, when possible, potency and function (measured by dynamic glucose-stimulated insulin secretion measurements and transplants in immunodeficient B6 Rag(+/-) mice). Postshipment OCR/DNA was not reduced in HD vs LD and was substantially reduced in the NC condition. HD islets exhibited normal function postshipment. Based on the data, we conclude that entire islet isolations (up to 400,000 IE) may be shipped using a single, larger SRM vessel with no negative effect on viability and ex vivo and in vivo function.

The DEXLIFE study methods: identifying novel candidate biomarkers that predict progression to type 2 diabetes in high risk individuals.

The incidence of type 2 diabetes (T2D) is rapidly increasing worldwide and T2D is likely to affect 592 million people in 2035 if the current rate of progression is continued. Today, patients are diagnosed with T2D based on elevated blood glucose, either directly or indirectly (HbA1c). However, the information on disease progression is limited. Therefore, there is a need to identify novel early markers of glucose intolerance that reflect the underlying biology and the overall physiological, metabolic and clinical characteristics of progression towards diabetes. In the DEXLIFE study, several clinical cohorts provide the basis for a series of clinical, physiological and mechanistic investigations in combination with a range of--omic technologies to construct a detailed metabolic profile of high-risk individuals across multiple cohorts. In addition, an exercise and dietary intervention study is conducted, that will assess the impact on both plasma biomarkers and specific functional tissue-based markers. The DEXLIFE study will provide novel diagnostic and predictive biomarkers which may not only effectively detect the progression towards diabetes in high risk individuals but also predict responsiveness to lifestyle interventions known to be effective in the prevention of diabetes.

The osteogenic potential of the marine-derived multi-mineral formula aquamin is enhanced by the presence of vitamin D.

Bone degenerative diseases are on the increase globally and are often problematic to treat. This has led to a demand to identify supplements that aid bone growth and formation. Aquamin is a natural multi-mineral food supplement, derived from the red algae Lithothamnion species which contains calcium, magnesium and 72 other trace minerals. It has been previously reported to increase bone formation and mineralisation. This study aimed to investigate the 28 day in vitro osteogenic response of Aquamin supplemented with Vitamin D. The osteogenic potential of MC3T3-E1 osteoblast-like cells was analysed in standard osteogenic medium supplemented with Aquamin +/- Vitamin D3, and the controls consisted of osteogenic medium, +/- Vitamin D3. Proliferation of osteoblasts, metabolic activity and cell viability did not differ between Aquamin and the osteogenic control groups. Alkaline phosphatase (ALP) levels and mineralisation were increased by the supplementation of Aquamin, and the addition of Vitamin D3 increased mineralisation for all groups. The combination of Aquamin and Vitamin D3 yielded a significant increase in ALP and mineralisation over Aquamin alone and the standard osteogenic control +/- Vitamin D3. This study demonstrates that Aquamin aids osteogenesis, and that its osteogenic response can be enhanced by combining Aquamin with Vitamin D3.

Grapevine Trunk Diseases in British Columbia: Incidence and Characterization of the Fungal Pathogens Associated with Black Foot Disease of Grapevine.

Black foot disease of grapevines, caused by several fungal species in the genera Campylocarpon, Cylindrocarpon, Cylindrocladiella, and Ilyonectria, causes significant economic losses to the grapevine industry worldwide. This study represents the first attempt to identify and characterize the fungal pathogens associated with black foot disease of grapevines in British Columbia (BC). Field surveys conducted throughout all grape-growing regions in BC that included assessment of foliar symptomatology and isolations from symptomatic vines showed Cylindrocarpon/Ilyonectria spp. occurred in 32 of 90 (35.5%) young vineyards surveyed (≤8 year old) and in 41 of 215 (19%) samples collected. In 20 of the 41 (48.8%) samples, Cylindrocarpon/Ilyonectria spp. were the sole fungi isolated from symptomatic tissue. In the rest of the samples, black foot fungi were found to primarily coexist with fungal taxa associated with Petri disease of grapevines. Colony and conidia phenotypical characterization, along with DNA analyses of the internal transcribed spacer region (ITS1-5.8S-ITS2) of the rDNA, and part of the ß-tubulin and translation elongation factor 1-α genes, revealed five different black foot fungi occurring in declining young vines in BC, namely Cylindrocarpon pauciseptatum, Ilyonectria liriodendri, Ilyonectria macrodidyma, Ilyonectria robusta, and Ilyonectria torresensis. Pathogenicity studies showed all five species to be highly virulent in the grapevine rootstock cultivar 3309C. Overall, I. liriodendri and I. macrodidyma were the most virulent species when inoculated in Vitis vinifera 'Chardonnay' and rootstock 3309C.

Grapevine Trunk Diseases in British Columbia: Incidence and Characterization of the Fungal Pathogens Associated with Esca and Petri Diseases of Grapevine.

Esca and Petri disease are two economically important grapevine diseases worldwide. This study reports for the first time the occurrence of both diseases on grapevines in British Columbia (BC) and subsequently in Canada. Visual assessment of 55,699 vines in 118 vineyards revealed a low incidence of esca with only 104 (0.2%) vines showing foliar symptoms. Young vine decline (YVD) was observed in 1,910 (7.8%) of 24,487 monitored young vines and in 52 (8%) of 654 young vines used as re-plants in mature vineyards. In 8 of 51 monitored young vineyards, YVD-affected vines ranged between 15 and 55%. Morphological studies along with DNA analyses of the ITS1-5.8S-ITS2, and part of the ß-tubulin, actin, and translation elongation factor 1-α gene regions, allowed us to identify Cadophora luteoolivacea, Phaeomoniella chlamydospora, Phaeoacremonium iranianum, Togninia fraxinopennsylvanica, Togninia minima, and the novel species Phaeoacremonium canadense and Phaeoacremonium roseum from esca and Petri disease infected vines in BC. This study includes for the first time the EF1-α DNA marker in Phaeoacremonium spp. delineation. Pathogenicity studies showed all seven fungi to cause vascular symptoms similar to those observed in esca and Petri disease infected vines. Additionally, the "tiger-stripes" foliar symptom of esca was successfully reproduced when healthy potted vines were inoculated with BC isolates of Pa. chlamydospora, Pm. canadense, Pm. iranianum, T. fraxinopennsylvanica, and T. minima.

The influence of a magnesium-rich marine extract on behaviour, salivary cortisol levels and skin lesions in growing pigs.

Growing pigs can display undesirable behaviours, reflecting or causing poor welfare. Addition of magnesium (Mg) to the diet could reduce these, as Mg supplementation has been associated with improved coping ability in response to stress. This study examined the effect of supplementation with a Mg-rich marine extract-based product (Supplement) on the behaviour, skin and tail lesion scores and salivary cortisol concentrations of growing pigs. At weaning (28 days), 448 piglets were assigned to either Control or Supplement (0.05%) diets in single-sex groups of 14. Four weeks later (c. 17 kg), pigs were blocked according to weight and back test scores. Seven piglets from each pen were mixed with seven from another pen of the same sex and dietary treatment to yield the following groups: control male, Supplement male, control female and Supplement female (n = 4 of each). This marked the start of the 9-week experimental period. Instances of the following behaviours were recorded in each pen for 8 × 2 min periods 1 day/week: aggression (fight, head-knock and bite); harmful (tail-in-mouth, ear-chewing and belly-nosing); and sexual/mounting behaviour. Four focal pigs were selected from each pen, and their behaviour was continuously recorded for 2 × 5 min periods on the same day. Saliva was collected once per week at 1000 h by allowing pigs to chew on a cotton bud for c. 1 min. Salivary cortisol was analysed in duplicate by an enzyme immunoassay. Skin and tail lesions were scored according to severity 1 day/week. There were fewer aggressive incidents in Supplement pens (P < 0.01), and mounting behaviour (performed only by males) was almost three times lower in Supplement than in control pens (P < 0.01). However, there was no effect of Supplement on the incidence of each of the harmful behaviours. Behaviour of the focal pigs showed no treatment effect on the duration or incidence of aggressive behaviour. However, Supplement pigs spent less time performing harmful behaviours compared with control pigs (P < 0.001). Supplement had no effect on the occurrence or severity of tail-biting outbreaks or on tail lesion scores. However, Supplement females had lower skin lesion scores, in particular in the ears and shoulders (P < 0.01). Finally, Supplement pigs had lower salivary cortisol concentrations (P < 0.01). Mounting is a major welfare concern in uncastrated pigs, and therefore this represents an important welfare benefit of Supplement. Reduced salivary cortisol, in conjunction with reduced skin lesion scores in supplemented females, suggests that addition of a Mg-rich marine extract improved pig welfare in this system.

Human islet distribution program for basic research at a single center.

The Clinical Islet Laboratory at the University of Alberta/Alberta Health Services distributes human islets for basic research when islet preparations fail to meet defined release criteria for transplantation. This report highlights our islet distribution activity for diabetes research over a 3-year period. Shipments of the acinar-enriched fraction for research were not included in this report. In 2010, we distributed 6.3 million islet equivalents (IEQs) of islets through 127 shipments to 8 researchers, locally, nationally, and internationally. The number of preparations for research use was stable over the 3-year period (26, 23, and 29 preparations in 2008, 2009, and 2010, respectively). Islet yield distributed for research per isolation was 201, 212, and 218 × 10(3) IEQs, respectively. The number of basic researchers was stable as well, although there were only 2 researchers before 2007. Recently, each researcher has received fewer islets per shipment (49,820 IEQs in 2010 vs 75,635 IEQs in 2008) but more frequently (21.5 in 2010 vs 11.2 times per year in 2008). This paradigm shift would be desirable for researchers, because in our experience, most require <30,000 IEQs per shipment, and more frequent islet shipments results in a larger sample size for experimentation. After an initial expansion in the number of researchers requesting islets, our islet distribution activity has remained stable over the years in terms of total productivity of islets utilized for research. The current supply-versus-demand ratio in our program appears to be appropriate.

Haplotype association of calpain 10 gene variants with type 2 diabetes mellitus in an Irish sample.

BACKGROUND: Calpain 10 (CAPN10) gene may contribute to the pathogenesis of type 2 diabetes mellitus (T2DM). AIM: To examine the contribution of four CAPN10 gene variants to T2DM risk in an Irish sample. METHODS: Genotyping of marker 19 insertion-deletion (ins/del) and three CAPN10 variants, rs3792267, rs3749166 and rs5030952 at the CAPN10 gene was performed in 236 T2DM subjects and 120 controls. Allelic, genotypic and haplotype comparisons were conducted between the groups. RESULTS: In the examined markers, no significant differences were observed although the deletion/deletion allele tended to be more common in T2DM subjects (chi(2) = 3.2, P = 0.07). A significant overrepresentation of a haplotype comprising (rs3792267), (19) and rs3749166 (chi(2) = 5.3, P = 0.021) was seen in T2DM subjects. Two protective haplotypes were detected: (G-ins-G) of (rs3792267), (19) and rs3749166 (chi(2) = 6.7, P = 0.009) and (ins-G-C) of (19), (rs3749166) and rs5030952 (chi(2) = 8.5, P = 0.003). CONCLUSIONS: CAPN10 gene variants may affect T2DM susceptibility in the Irish population.

First Report of a New Postharvest Disease of Pear Fruit Caused by Sphaeropsis pyriputrescens in Canada.

A survey of stored d'Anjou pears was conducted in British Columbia (BC), Canada in January 2006 to determine if Sphaeropsis rot was present in BC as had been reported previously for apples and pears in Washington (1,2). Sphaeropsis pyriputrescens Xiao & J.D. Rogers produces decay similar to Botrytis cinerea that originates from the stem or calyx end. Of 3,614 pears sampled, 55 (1.5%) had symptoms similar to those described for Sphaeropsis rot. Isolations were made from each infected pear onto acidified potato dextrose agar (APDA) dishes and incubated at 20°C for 5 to 7 days. Twenty-seven cultures resembling S. pyriputrescens were induced to produce pycnidia by exposing them to 12-h cycles of alternating light and dark periods at 20°C (1). Conidia extracted from pycnidia were then streaked onto PDA dishes and incubated at 20°C for 12 to 24 h from which single-spore cultures were made. These isolates developed a dense, white-to-cream mycelium that turned yellow over time; black pycnidia were formed on the culture dishes after 4 weeks. Conidia were brown, clavate to subglobose to irregular, and similar in size (16 × 10 µm) to previous descriptions (1). Identification of S. pyriputrescens was confirmed by using DNA sequence data from the ß-tubulin and ribosomal genes. Sequences from S. pyriputrescens from Washington (1) were compared with those from BC, Canada. Isolates from Canada shared 99 to 100% sequence homology with those from Washington. Two of the BC isolates (DAOM 238917 and 238918) were deposited in the Canadian Culture Collection, Ottawa, ON and their corresponding sequences were placed in the GenBank database (NCBI, Bethesda, MD) with accession nos. EU156037 and EU156040 (ribosomal gene) and EU156048 and EU 156050 (ß-tubulin gene), respectively. Five isolates from different locations in BC and two isolates from Washington were tested for pathogenicity on d'Anjou pears and four apple cultivars (Ambrosia, Fuji, Gala, and Granny Smith). Plugs (3 mm in diameter) removed from 2-week-old cultures were placed into two corresponding wounds on each of five fruit per cultivar. The fruit were then placed at 1 or 20°C for 22 or 7 days, respectively, when the diameters of the decay areas were recorded. All isolates were pathogenic on pears (P = <0.05). Decay lesion diameter was greater at 1°C, ranging from 46.8 to 57.9 mm, than at 20°C, ranging from 32.6 to 44.2 mm. All BC isolates were also pathogenic on the fruit of each apple cultivar (P = <0.05), although at 20°C, decay areas were smaller than on pears, and at 1°C, very little rot developed. Koch's postulates were completed by reisolating S. pyriputrescens from the inoculated pears and apples and identifying the isolates as above. Although S. pyriputrescens was only observed on pears in BC, research in Washington indicates that it is a more serious problem on apples (2). To our knowledge, this is the first documented report of the occurrence of S. pyriputrescens in Canada. References: (1) C. L. Xiao and J. D. Rogers. Plant Dis. 88:114, 2004. (2) C. L. Xiao et al. Plant Dis. 88:223, 2004.

Exercise training increases insulin-stimulated glucose disposal and GLUT4 (SLC2A4) protein content in patients with type 2 diabetes.

AIMS/HYPOTHESIS: Exercise enhances insulin-stimulated glucose transport in skeletal muscle through changes in signal transduction and gene expression. The aim of this study was to assess the impact of acute and short-term exercise training on whole-body insulin-mediated glucose disposal and signal transduction along the canonical insulin signalling cascade. METHODS: A euglycaemic-hyperinsulinaemic clamp, with vastus lateralis skeletal muscle biopsies, was performed at baseline and 16 h after an acute bout of exercise and short-term exercise training (7 days) in obese non-diabetic (n=7) and obese type 2 diabetic (n=8) subjects. RESULTS: Insulin-mediated glucose disposal was unchanged following acute exercise in both groups. Short-term exercise training increased insulin-mediated glucose disposal in obese type 2 diabetic (p<0.05), but not in obese non-diabetic subjects. Insulin activation of (1) IRS1, (2) IRS2, (3) phosphotyrosine-associated phosphatidylinositol-3 kinase activity and (4) the substrate of phosphorylated Akt, AS160, a functional Rab GTPase activating protein important for GLUT4 (now known as solute carrier family 2 [facilitated glucose transporter], member 4 [SLC2A4]) translocation, was unchanged after acute or chronic exercise in either group. GLUT4 protein content was increased in obese type 2 diabetic subjects (p<0.05), but not in obese non-diabetic subjects following chronic exercise. CONCLUSIONS/INTERPRETATION: Exercise training increased whole-body insulin-mediated glucose disposal in obese type 2 diabetic patients. These changes were independent of functional alterations in the insulin-signalling cascade and related to increased GLUT4 protein content.

Multi-lot analysis of custom collagenase enzyme blend in human islet isolations.

INTRODUCTION: Variability currently in Liberase HI from lot to lot limits the ability to effectively isolate islets with consistency. Roche Diagnostics Inc (Indianapolis, Ind, USA) has developed a Custom Collagenase enzyme blend in hopes that producing collagenase II and I and thermolysin separately will eliminate variability. In this study we examined the variability in Custom Collagenase lots in respect to isolation results and isolation success rates and compared those to Liberase HI. METHODS: We retrospectively analyzed records from 68 islet isolations where either Liberase HI (lot A: n = 23, Lot B: n = 20) or Custom Collagenase blend (Lot C: n = 10, Lot D: n = 15) was employed. Human islets were isolated from cadaveric pancreata using standardized methods performed in a controlled islet isolation facility. RESULTS: Analysis of Liberase HI and Custom Collagenase using Student t test showed no difference between the two groups. Comparison of the two Custom Collagenase lots using the t test showed a statistical difference between undigested pancreas weight and pancreas digestion times. Using chi-square test, no statistical significance was found in isolation success rates from lot to lot. CONCLUSION: Although the Custom Collagenase blend is comparable to Liberase HI in its ability to isolate human islets, variability still exists from lot to lot when used conventionally as Liberase HI is. The ability to predetermine doses is beneficial, and as techniques to manipulate the activity levels prior to isolations improve so to will the enzymes' ability to isolate islets on a consistent basis.