Production of high fructose syrup from asparagus inulin using immobilized exoinulinase from Kluyveromyces marxianus YS-1

Extracellular exoinulinase from Kluyveromyces marxianus YS-1, which hydrolyzes inulin into fructose, was immobilized on Duolite A568 after partial puriWcation by ethanol precipitation and gel exclusion chromatography on Sephadex G-100. Optimum temperature of immobilized enzyme was 55 °C, which was 5 °C higher than the free enzyme and optimal pH was 5.5. Immobilized biocatalyst retained more than 90% of its original activity after incubation at 60 °C for 3 h, whereas in free form its activity was reduced to 10% under same conditions, showing a signiWcant improvement in the thermal stability of the biocatalyst after immobilization. Apparent Km values for inulin, raYnose and sucrose were found to be 3.75, 28.5 and 30.7 mM, respectively. Activation energy (Ea) of the immobilized biocatalyst was found to be 46.8 kJ/mol. Metal ions like Co2+ and Mn2+ enhanced the activity, whereas Hg2+ and Ag2+ were found to be potent inhibitors even at lower concentrations of 1 mM. Immobilized biocatalyst was eVectively used in batch preparation of high fructose syrup from Asparagus racemosus raw inulin and pure inulin, which
yielded 39.2 and 40.2 g/L of fructose in 4 h; it was 85.5 and 92.6% of total reducing sugars produced, respectively.

Source of Bet v 1 loaded inhalable particles from birch revealed

Allergenic proteins present in pollen grains, when inhaled, interact with the airways to cause an attack of asthma in susceptible humans. In one system, grass pollen grains rupture osmotically in rainfall, releasing allergen-containing inhalable particles into the atmosphere. In contrast, birch tree pollen grains do not rupture under these conditions, yet the major allergen, Bet v 1, has been detected in the atmosphere in inhalable particles of unknown origin. It is possible that Bet v 1 may diffuse from intact settled pollen grains and the allergenic material may again become airborne, interacting with settled fine particles from other sources prior to resuspension. This study investigates the mechanism for the release of birch pollen allergen-containing inhalable particles from pollen grains. We propose the hypothesis that (1) airborne birch pollen grains settle on nearby leaf surfaces; (2) then, following light rainfall, the grains germinate and, (3) later, pollen tubes burst, releasing inhalable particles carrying Bet v 1 into the atmospheric aerosol.   We used microscopic analyses of pollen behaviour following anther opening, a Burkard volumetric trap for pollen counts and a high volume air sampler with a two-stage cascade impactor for quantitative immunochemical analyses of Bet v 1. On dry days of high birch pollen count (48 grains/m3, 1.5 ng/m3 of Bet v 1), we found that the surfaces of birch leaves became coated with pollen. This ”pollen rain” is a source of secondary emission of allergens into the atmosphere. We observed that following light rainfall (<1 mm per day), about 80% of the birch pollen grains germinated, producing pollen tubes, especially in the sticky surface secretions of leaf glands. These pollen tubes may grow up to 300 μm in length prior to rupturing, each releasing about 400 starch granules coated with allergen molecules that may, after drying, be dispersed into the aerosol. On these days following light rainfall, the highest atmospheric levels of Bet v 1 (1.18 ng/m3) are associated with inhalable particles. Following heavy rainfall, both pollen and inhalable particles are washed from the atmosphere. Immunoprinting studies show that Bet v 1 is associated with starch granules rather than the smaller orbicules. Bet v 1 is present in the atmosphere in large particles, i.e. in particular pollen grains and in inhalable particles, i.e. in particular starch granules.

Dietary salt loading impairs arterial vascular reactivity 1-3

BACKGROUND: Studies of sodium have shown improvements in vascular function and blood pressure (BP). The effect of chronic sodium loading from a low-sodium diet to a Western diet on vascular function and BP has been less well studied.

OBJECTIVE: The objective was to examine the effects of dietary salt intake on vascular function and BP.

DESIGN: Thirty-five hypertensive volunteers met the inclusion criteria. After a 2-wk run-in with a low-sodium diet (60 mmol/d), the participants maintained their diets and were randomly assigned to receive sequentially 1 of 3 interventions for 4 wk, with a 2-wk washout between interventions: sodium-free tomato juice (A), tomato juice containing 90 mmol Na (B), and tomato juice containing 140 mmol Na (C). The outcomes were changes in pulse wave velocity (PWV), systolic BP (SBP), and diastolic BP (DBP).

RESULTS: The difference in PWV between interventions B and A was 0.39 m/s (95% CI: 0.18, 0.60 m/s; P = 0.001) and between C and A was 0.35 m/s (95% CI: 0.13, 0.57 m/s; P = 0.01). Differences in SBP and DBP between interventions B and A were 4.4 mm Hg (95% CI: 1.2, 7.8 mm Hg; P = 0.01) and 2.4 mm Hg (95% CI: 0.8, 4.1 mm Hg; P = 0.001), respectively, and between interventions C and A were 5.6 mm Hg (95% CI: 2.7, 8.4 mm Hg; P = 0.01) and 3.3 mm Hg (95% CI: 1.5, 5.0 mm Hg; P = 0.001), respectively. Changes in PWV correlated with changes in SBP (r = 0.52) and DBP (r = 0.58).

CONCLUSIONS: Dietary salt loading produced significant increases in PWV and BP in hypertensive volunteers. Correlations between BP and PWV suggest that salt loading may have a BP-independent effect on vascular wall function. This further supports the importance of dietary sodium restriction in the management of hypertension. This trial was registered with the Australian and New Zealand Clinical Trials Registry as ACTRN12609000161224.

Growth rates of the infaunal bivalve Soletellina alba (Lamarck, 1818) (Bivalvia: Psammobiidae) in an intermittent estuary of southern Australia

Caging and a mark–recapture design were used to estimate the growth rate of the brittle, infaunal bivalve Soletellina alba in the Hopkins River estuary. The growth of both caged and uncaged individuals was monitored at three sites near the mouth of the estuary over 180 days. Growth rates did not differ for caged and uncaged bivalves, or for bivalves subject to different amounts of handling, or between sites. Growth did differ between consecutive time intervals, which was attributable to negligible growth occurring during the colder months of autumn/winter. Comparisons of the condition (as indicated by total mass for length³) of S. alba were inconsistent between sites for caged and uncaged bivalves and for those subject to different amounts of handling. Soletellina alba is a rapidly growing bivalve with mean growth rates for the three time intervals being 0.04±0.002 mm day⁻¹ in summer, 0.02±0.001 mm day⁻¹ in autumn and 0.03±0.001 mm day⁻¹ from summer to winter. Using existing literature, it was shown that a significant relationship exists between maximum shell length and onset of sexual maturity in bivalve molluscs. This relationship predicts that S. alba should reach the onset of sexual maturity at 15.8 mm length. Therefore, it appears that it may be possible for juvenile S. alba (<1 mm) to grow, reach sexual maturity and reproduce in between annual mass-mortality events caused by winter flooding.

Caffeine as a flavor additive in soft-drinks

Over 60% of soft-drinks sold in the United States contain caffeine, a mildly addictive psycho-active chemical, as a flavor additive. Using sweeteners as controls, we assessed whether caffeine has flavor activity in a cola soft-drink. A forced-choice triangle discrimination methodology was used to determine detection thresholds of caffeine in sweeteners and a cola beverage. The subjects (n=30, 28 female, 23±4 years old) were trained tasters and completed over 1600 discrimination tests during the study. The mean detection thresholds for caffeine in the sweet solutions were: 0.333±0.1 mM sucrose; 0.467±0.29 mM aspartame; 0.462±0.3 mM sucralose, well below the concentration in common cola beverages (0.55–0.67 mM). A fixed concentration of caffeine, corresponding to the concentration of caffeine in a common cola beverage (0.67 mM) was added to the sweeteners and a non-caffeinated cola beverage. Subjects could distinguish between caffeinated and non-caffeinated sweeteners (p<0.001), but all subjects failed to distinguish between caffeinated and non-caffeinated cola beverage (p=1.0). Caffeine has no flavor activity in soft-drinks yet will induce a physiologic and psychologic desire to consume the drink.

Spatial and temporal changes in abundance of the infaunal bivalve Soletellina alba (Lamark, 1818) during a time of drought in the seasonally-closed Hopkins River Estuary, Victoria Australia

The infaunal bivalve Soletellina alba is susceptible to mass mortalities during annual winter flooding in the Hopkins River Estuary, southern Australia. Periods of low salinity (≤1) are the likely cause of these mass mortality events, which can occur in seasonally-closed estuaries when high winter flows are sufficient to flush all salt water from the estuary. Core samples of S. alba were collected from two water depths across four times and at three sites near the mouth of the estuary. Minimal to zero abundances of large S. alba (>1 mm) were expected to be sampled, particularly at the shallower water depth, during a typical winter flood event. However, the present study occurred during a period of drought, which led to the absence of winter flooding. This absence of winter flooding prevented the occurrence of lethal salinities (i.e. ≤1) in the estuary during this period and a greater number of living S. alba adults were sampled. Abundances of juvenile and adult S. alba were still variable, even in the absence of winter flooding, and reflected an interaction between date, site and water depth. However, no mass mortalities of adults were observed during the drought conditions in contrast to what occurs during typical winter flood events and provides support for the hypothesis that winter flooding is responsible for past mass mortalities.

Experimental study on effect of loading rate on mode I delamination of z-pin reinforced laminates

This paper presents an experimental investigation on mode I delamination of z-pinned double-cantilever-beams (DCB) and associate z-pin bridging mechanisms. Tests were performed with three types of samples: big-pin with an areal density of 2%, small-pin with an areal density of 2% and small-pin with an areal density of 0.5%. The loading rates for each type of samples were set at 1 mm/min and 100 mm/min. Comparison of fracture load under different loading rates shows the rate effects on delamination crack opening and delamination growth. Optical micrographs of z-pins after pullout were also presented to identify the bridging mechanisms of z-pins under different loading rates.

The effect of artificial defects on the compressive properties of hollow sphere cellular aluminium (HSCA)

Hollow sphere cellular aluminium (HSCA) samples were fabricated by bonding together two kinds of single aluminium hollow spheres with the same outside diameter of 4 mm but different wall thicknesses of 0.1 mm and 0.3 mm, in which the hollow spheres with the thinner sphere wall thickness were used as artificial defects. Four types of HSCA samples with the same relative density but various distributions of artificial defects were prepared by simple cubic packing. For comparing, HSCA sample without defective hollow spheres inside was also prepared. The effects of the distribution of the artificial defects on the deformation behaviours and mechanical properties were investigated by compressive tests. Results indicated that the nominal stress - nominal strain curve and the deformation behavior of the HSCA samples varied with the distribution of the artificial defects in spite of the same relative density. It is therefore suggested that the deformation behavior and mechanical property of cellular materials were also significantly affected by the distribution of defects. In particular, the plateau stress of the HSCA samples increased with the decrease in number of contact points between the normal hollow spheres and the defective hollow spheres in the loading direction during deformation.

Compressive properties and energy absorption of hollow sphere aluminium

Manufactured cellular aluminums have been developed for a wide range of automotive applications where weight savings, improved safety, crashworthiness and comfort are required. The plateau deformation behavior of cellular aluminums under compressive loading makes this new class of lightweight materials suitable for energy absorption and comes close to ideal impact absorbers. In the present study, aluminum hollow hemispheres were firstly processed by pressing. Hollow sphere aluminum samples with a body-centered cubic (BCC) packing were then fabricated by bonding together single hollow spheres, which were prepared by adhering together hollow hemispheres. Hollow sphere aluminum samples with various kinds of sphere wall thicknesses of 0.1 mm, 0.3 mm and 0.5 mm but the same outside diameter of 4 mm were investigated by compressive tests. The effects of the sphere wall thickness on the mechanical properties and energy absorption characteristics were investigated.

Processing and mechanical properties of hollow sphere aluminum foams

Hollow sphere metallic foams are a new class of cellular material that possesses the attractive advantages of uniform cell size distribution and regular cell shape. These result in more predictable physical and mechanical properties than those of cellular materials with a random cell size distribution and irregular cell shapes. In the present study, single aluminum hollow spheres with three kinds of sphere wall thickness as 0.1 mm, 0.3 mm and 0.5 mm were processed by a new pressing method. Hollow sphere aluminum foam samples were prepared by bonding together single hollow spheres with simple cubic packing (SC) and body-centered cubic packing (BCC). Compressive tests were carried out to evaluate the deformation behaviors and mechanical properties of the hollow sphere aluminum foams. Effects of the sphere wall thickness and packing style on the mechanical properties were investigated.

Population dynamics of the infaunal bivalve, Soletellina alba

The population dynamics of the infaunal bivalve Soletellina alba was investigated at three sites situated within close proximity to the mouth of the Hopkins River estuary. The initial study design was planned to examine the importance of winter flooding to the persistence of this bivalve mollusc within the Hopkins estuary, since mass mortalities have been observed during previous years coincident with periods of winter flooding. Unfortunately, the climatic conditions experienced during this study were atypical compared to the long-term average, so detailed sampling was limited to two, unanticipated, non-flood years rather than two, highly anticipated, flood years. This hampered my ability to conduct complete tests of the importance of winter flooding. Patterns of river discharge and the frequency and duration of mouth opening and closing differed greatly from that expected. Unexpectedly, periods of mouth closure were not always associated with periods of minimal river discharge; low salinities were another unexpected result during an extended period of mouth closure during 1998. As expected, salinities varied considerably with increasing water depth when the estuary mouth was open. Mouth closure lead to salinities becoming more uniform between water depths but hypoxic and anoxic conditions became evident via stratification in the water column at 1 m below the Australian Height Datum (AHD). Other than trends associated with increased water depth, significant variation was not evident between measurements of salinity taken from three sites within close proximity of the estuary mouth (approximately 500 m), or during changes in tide. The most pertinent anomaly was the absence of winter flooding. The distribution and abundance of juvenile and adult S. alba was variable across all Dates, Sites and Channel elevations (i.e. water depths) sampled during this study. An experimental test comparing the recruitment of juveniles at different channel elevations and in sediments of varying particle size was conducted during an exceptionally successful period of recruitment during 1999. The results of these tests showed that recruitment was greatest at the shallowest channel elevation used, and there was little evidence that sediment particle size influenced recruitment. In contrast to 1999, recruitment during 1997 or 1998 was extremely poor. Growth rates were monitored using tagged individuals held in caged and uncaged plots, which revealed that growth was highly variable among individuals, but not between Sites. These tests also revealed that growth was negligible during the colder, winter months, and that the fastest growing individuals were capable of growing 0.2 mm/day. Mixed results were obtained for tests of potential cage artifacts and the influence of handling. Caging and differing amounts of handling did not appear to influence growth, but there was evidence that cages and handling influenced bivalve condition and number of mortalities. These direct tests appeared to be the most appropriate method for determining growth rates of this species, since attempts to analyse length-frequency data were made difficult by the apparent convergence of cohorts, and shell aging is difficult due to the thin, fragile nature of the shell. As expected, mass mortalities were observed during the flood of 1996, but not during the two non-flood years of 1997 and 1998. There were, however, some considerable declines in abundances at some channel elevations during the two non-flood years. However, these declines were attributable to the complete disappearance of individuals, rather than the sudden presence of numerous, recently dead individuals that typify observed declines during winter flooding. The complete disappearance of individuals suggest that S. alba may be capable of post-settlement emigration, or that they were consumed by an unknown predator. Salinity tolerance tests showed that bivalves exposed to low salinities (≤6 ppt), exhibited poorer condition and took longer to re-burrow into sediments than those exposed to greater salinities (≥14 ppt), while death of bivalves exposed to salinities ≤1 ppt occurred after 8 days of exposure. These tests provide evidence that low salinities are probably the principal cause of mass mortalities during winter flooding, although the interaction between salinity, temperature and turbidity also deserve consideration. The results of this study indicate that certain aspects of winter flooding, especially salinity, are responsible for the mass mortalities of S. alba rather than the result of a short-lived life history. I hypothesise that the survival of very young juveniles (between 0.5 and 1 mm shell length) and rapid growth rates are important features of the life history of S. alba that explain its successful persistence within the Hopkins River estuary. The rapid rates of growth suggest that it may be possible for juveniles that survive winter flooding to grow, reach sexual maturity, and reproduce before the onset of the next flood event. Unfortunately, the increased survivorship of juveniles during periods of winter flooding was not demonstrated by this study because of the absence of winter flooding and also relatively poor recruitment. It is highly likely that this species is capable of completing it entire life cycle within the estuary since the absence of other nearby populations, together with periods of mouth closure, are likely to greatly limit the potential contribution made by larvae entering from the surrounding marine environment. This study has added considerably to our knowledge of how infauna cope with life in the intermittently closing estuaries that typify semi-arid coastlines in the Southern Hemisphere.

Familial and genomic analyses of postural changes in systolic and diastolic blood pressure

The physiological adaptation to the erect posture involves integrated neural and cardiovascular responses that might be determined by genetic factors. We examined the familial- and individual-specific components of variance for postural changes in systolic and diastolic blood pressure in 767 volunteer nuclear adult families from the Victorian Family Heart Study. In 274 adult sibling pairs, we made a genome-wide scan using 400 markers for quantitative trait loci linked with the postural changes in systolic and diastolic pressures. Overall, systolic pressure did not change on standing, but there was considerable variation in this phenotype (SD=8.1 mm Hg). Familial analyses revealed that 25% of the variance of change in systolic pressure was attributable to genetic factors. In contrast, diastolic pressure increased by 6.3 mm Hg (SD=7.0 mm Hg) on standing and there was no evidence of contributory genetic factors. Multipoint quantitative genome linkage mapping suggested evidence (Z=3.2) of linkage of the postural change in systolic pressure to chromosome 12 but found no genome-wide evidence of linkage for the change in diastolic pressure. These findings suggest that genetic factors determine whether systolic pressure decreases or increases when one stands, possibly as the result of unidentified alleles on chromosome 12. The genetics of postural changes in systolic blood pressure might reflect the general buffering function of the baroreflex; thereby, the predisposition to sudden decreases or increases in systolic pressure might cause postural hypotension or vessel wall disruption, respectively.

Computer-assisted image analysis of liver collagen: relationship to Ishak scoring and hepatic venous pressure gradient

Histopathological scoring of disease stage uses descriptive categories without measuring the amount of fibrosis. Collagen, the major component of fibrous tissue, can be quantified by computer-assisted digital image analysis (DIA) using histological sections. We determined relationships between DIA, Ishak stage, and hepatic venous pressure gradient (HVPG) reflecting severity of fibrosis. One hundred fifteen patients with hepatitis C virus (HCV) who had undergone transplantation had 250 consecutive transjugular liver biopsies combined with HVPG (median length, 22 mm; median total portal tracts, 12), evaluated using the Ishak system and stained with Sirus red for DIA. Liver collagen was expressed as collagen proportionate area (CPA). Median CPA was 6% (0.2-45), correlating with Ishak stage (stage 6 range, 13%-45%), and with HVPG (r = 0.62; P < 0.001). Median CPA was 4.1% when HVPG was less than 6 mm Hg and 13.8% when HVPG was 6 mm Hg or more (P < 0.0001) and 6% when HVPG was less than 10 mm Hg and 17.3% when HVPG was 10 mm Hg or higher (P < 0.0001). Only CPA, not Ishak stage/grade, was independently associated by logistic regression, with HVPG of 6 mm Hg or more [odds ratio, 1.206; 95% confidence interval (CI), 1.094-1.331; P < 0.001], or HVPG of 10 mm Hg or more (odds ratio, 1.105; 95% CI, 1.026-1.191; P = 0.009). CPA increased by 50% (3.6%) compared with 20% in HVPG (1 mm Hg) in 38 patients with repeated biopsies. Conclusion: CPA assessed by DIA correlated with Ishak stage scores and HVPG measured contemporaneously. CPA was a better histological correlate with HVPG than Ishak stage, had a greater numerical change when HVPG was low, and resulted in further quantitation of fibrosis in cirrhosis.

Characterization of supported cylinder-planar germanium waveguide sensors with synchrotron Infrared radiation

Cylinder-planar Ge waveguides are being developed as evanescent-wave sensors for chemical microanalysis. The only non-planar surface is a cylinder section having a 300-mm radius of curvature. This confers a symmetric taper, allowing for direct coupling into and out of the waveguide's 1-mm² end faces while obtaining multiple reflections at the central <30-μm-thick sensing region. Ray-optic calculations indicate that the propagation angle at the central minimum has a strong nonlinear dependence on both angle and vertical position of the input ray. This results in rather inefficient coupling of input light into the off-axis modes that are most useful for evanescent-wave absorption spectroscopy. Mode-specific performance of the cylinder-planar waveguides has also been investigated experimentally. As compared to a blackbody source, the much greater brightness of synchrotron-generated infrared (IR) radiation allows a similar total energy throughput, but restricted to a smaller fraction of the allowed waveguide modes. However, such angle-selective excitation results in a strong oscillatory interference pattern in the transmission spectra. These spectral oscillations are the principal technical limitation on using synchrotron radiation to measure evanescent-wave absorption spectra with the thin waveguides.

High glucose-induced impairment in insulin secretion is associated with reduction in islet glucokinase in a mouse model of susceptibility to islet dysfunction

Type 2 diabetes is characterized by islet dysfunction resulting in hyperglycemia, which can then lead to further deterioration in islet function. A possible mechanism for hyperglycemia-induced islet dysfunction is the accumulation of advanced glycation end products (AGE). The DBA/2 mouse develops pancreatic islet dysfunction when exposed to a high glucose environment and/or obesity-induced insulin resistance. To determine the biochemical cause of dysfunction, DBA/2 and C57BL/6 control islets were incubated in 11.1 mM or 40 mM glucose in the absence or presence of the AGE inhibitor aminoguanidine (AG) for 10 days. Basal (2.8 mM glucose) insulin release was increased in both DBA/2 and C57BL/6 islets incubated with 40 mM vs 11.1 mM glucose for 10 days. Chronic exposure to hyperglycemia decreased glucose (20 mM)-stimulated insulin secretion in DBA/2 but not in C57BL/6 islets. AG significantly increased fold-induced insulin release in high glucose cultured DBA/2 mouse islets, but did not affect C57BL/6 islet function. DBA/2 islet glucokinase was significantly reduced following 40 mM glucose culture, compared with 11.1 mM glucose cultured DBA/2 islets and 40 mM glucose cultured C57BL/6 islets. Incubation of islets with AG resulted in a normalization of DBA/2 islet glucokinase levels. In conclusion, chronic high glucose-induced increases in AGE can result in islet dysfunction and this is associated with reduced glucokinase levels in a mouse model with susceptibility to islet failure.