Lactoylglutathione lyase, a critical enzyme in methylglyoxal detoxification, contributes to survival of Salmonella in the nutrient rich environment

Glyoxalase I which is synonymously known as lactoylglutathione lyase is a critical enzyme in methylglyoxal (MG) detoxification. We assessed the STM3117 encoded lactoylglutathione lyase (Lgl) of Salmonella Typhimurium, which is known to function as a virulence factor, due in part to its ability to detoxify methylglyoxal. We found that STM3117 encoded Lgl isomerises the hemithioacetal adduct of MG and glutathione (GSH) into S-lactoylglutathione. Lgl was observed to be an outer membrane bound protein with maximum expression at the exponential growth phase. The deletion mutant of S. Typhimurium (lgl) exhibited a notable growth inhibition coupled with oxidative DNA damage and membrane disruptions, in accordance with the growth arrest phenomenon associated with typical glyoxalase I deletion. However, growth in glucose minimal medium did not result in any inhibition. Endogenous expression of recombinant Lgl in serovar Typhi led to an increased resistance and growth in presence of external MG. Being a metalloprotein, Lgl was found to get activated maximally by Co2+ ion followed by Ni2+, while Zn2+ did not activate the enzyme and this could be attributed to the geometry of the particular protein-metal complex attained in the catalytically active state. Our results offer an insight on the pivotal role of the virulence associated and horizontally acquired STM3117 gene in non-typhoidal serovars with direct correlation of its activity in lending survival advantage to Salmonella spp.

Mg/BN nanocomposites: Nano-BN addition for enhanced room temperature tensile and compressive response

The present study elucidates the effects of nanoscale boron nitride particles addition on the microstructural and mechanical characteristics of monolithic magnesium. Novel light-weight Mg nanocomposites containing 0.3, 0.6 and 1.2vol% nano-size boron nitride particulates were synthesized using the disintegrated melt deposition method followed by hot extrusion. Microstructural characterization of developed Mg/x-boron nitride composites revealed significant grain refinement due to the uniform distribution of nano-boron nitride particulates. Texture analysis of selected Mg-1.2 boron nitride nanocomposite showed an increase in the intensity of fiber texture alongside enhanced localized recrystallization when compared to monolithic Mg. Mechanical properties evaluation under indentation, tension and compression loading indicated superior response of Mg/x-boron nitride composites in comparison to pure Mg. The uniform distribution of nanoscale boron nitride particles and the modified crystallographic texture achieved due to the nano-boron nitride addition attributes to the superior mechanical characteristics of Mg/boron nitride nanocomposites.

The physicochemical characteristics and anaerobic degradability of desiccated coconut industry waste water

Desiccated coconut industries (DCI) create various intermediates from fresh coconut kernel for cosmetic, pharmaceutical and food industries. The mechanized and non-mechanized DCI process between 10,000 and 100,000 nuts/day to discharge 6-150 m(3) of malodorous waste water leading to a discharge of 2646642 kg chemical oxygen demand (COD) daily. In these units, three main types of waste water streams are coconut kernel water, kernel wash water and virgin oil waste water. The effluent streams contain lipids (1-55 g/l), suspended solids (6-80 g/l) and volatile fatty acids (VFA) at concentrations that are inhibitory to anaerobic bacteria. Coconut water contributes to 20-50 % of the total volume and 50-60 % of the total organic loads and causes higher inhibition of anaerobic bacteria with an initial lag phase of 30 days. The lagooning method of treatment widely adopted failed to appreciably treat the waste water and often led to the accumulation of volatile fatty acids (propionic acid) along with long-chain unsaturated free fatty acids. Biogas generation during biological methane potential (BMP) assay required a 15-day adaptation time, and gas production occurred at low concentrations of coconut water while the other two streams did not appear to be inhibitory. The anaerobic bacteria can mineralize coconut lipids at concentrations of 175 mg/l; however; they are severely inhibited at a lipid level of = 350 mg/g bacterial inoculum. The modified Gompertz model showed a good fit with the BMP data with a simple sigmoid pattern. However, it failed to fit experimental BMP data either possessing a longer lag phase and/or diauxic biogas production suggesting inhibition of anaerobic bacteria.

Microstructure of Zr-alloyed coating using pulsed laser

Coatings were synthesized by laser alloying of zirconium (Zr) particles using a pulsed Nd:YAG laser on an austenite stainless steel. The distribution of Zr is uneven, in a depth of 18 mum just below the surface of the pool. The coating consisted of duplex microstructures, i.e. metallic glass (MG) and austenite. The MG formed in Zr-contained regions, with the Zr composition ranging from 7.6 to 16.8 at.%. The formation of the MG was attributed to an increase in glass-forming ability by Zr addition.

Impact of cathode evaporation on a free-burning arc

A free-burning, high-intensity argon arc at atmospheric pressure was modelled during the evaporation of copper from the cathode. The effect of cathode evaporation on the temperature, mass flow, current flow and Cu concentration was studied for the entire plasma region. The copper evaporates from the tip of the cathode with an evaporation rate of 1 mg s-1. The copper vapour in the cathode region has a velocity of 210 m s-1 with a mass concentration of above 90% within 0.5 mm from the arc axis. The vapour passes from the cathode toward the anode with a slight diffusion in the argon plasma. Higher temperatures and current densities were calculated in the core of the arc caused by the cathode evaporation.

Rapid determination of total CLA concentration in beef fat

[EN]Conjugated linoleic acid (CLA) has many potential healthful properties, and beef is naturally enriched with CLA. Simple and rapid methods to measure total CLA were investigated to enable sorting of beef carcasses with potential enhanced economic value. Direct alcohol extraction combined with measuring absorbance was simple, accurate and perhaps the most viable method for rapid carcass sorting compared to methods using saponification or methylation followed by extraction.

Subcutaneous fat composition of youthful and mature Canadian beef: emphasis on individual conjugated linoleic acid and trans-18:1 isomers

[EN]A comprehensive evaluation of the fatty acid composition of subcutaneous adipose tissue from beef cattle produced in western Canada was undertaken to determine if the current Canadian grading system is able to distinguish classes of animals with value added potential due to their fatty acid composition. Grades included youthful Canadian Yield Grade 1 A/AA beef, under (YUTM) and over (YOTM) 30 mo of age and the four mature grades (D1, D2, D2 and D4). Subcutaneous fat between the 12th and 13th ribs over the longissimus muscle was obtained from 18_21 animals per grade. Fatty acids were analyzed using a combination of silver-ion HPLC and GC with a highly polar 100 m column. There were no differences in total trans-18:1 content amongst grades, but adipose tissue from grade D1, D2 and D4 had more 11t-18:1 than YUTM (PB0.05), whereas adipose tissue from YUTM carcasses had more 10t-18:1 than all other grades (PB0.05). Adipose tissue from YUTM carcasses also had less total CLA (PB0.05) than the D grades, mainly due to a lower level of 9c,11t-CLA, but they had slightly more 7t,9c-CLA and 10t,12c-CLA (PB0.05). Adipose tissue from YOTM and D grades contained more n-3 fatty acids relative to YUTM (0.56% vs. 0.29%; PB0.05) and lower n-6:n-3 ratios (PB0.05). Overall, older animals (YOTM and D grades) had adipose tissue compositions with higher levels of fatty acids with reported health benefits. Taken together, these higher levels may provide opportunities for value added marketing if regulatory authorities allow claims for their enrichment based on demonstrated health benefits. Higher concentrations of beneficial fatty acids, however, need to be considered within the context of the complete fatty acid profile and it would be important to demonstrate their advantages in the presence of relatively high levels of saturated fatty acids.

Survey of the fatty acid composition of Canadian beef: Backfat and longissimus lumborum muscle

[EN]A survey of Canadian retail beef was undertaken with emphasis on the trans fatty acid (TFA) and conjugated linoleic acid (CLA) isomers, and compared with current health recommendations. Thirty striploin steaks were collected in the winter and summer from major grocery stores in Calgary (Alberta, Canada). Steak fatty acid compositions (backfat and longissimus lumborum muscle analysed separately) showed minor seasonal differences with lower total saturates (PB0.05) and higher total monounsaturates (PB 0.01) in winter, but no differences in total polyunsaturated fatty acids. The ratio of n-6 and n-3 polyunsaturated fatty acid in longissimus lumborum averaged 5.8. The average TFA content in longissimus lumborum was 0.128 g 100 g_1 serving size, and 10t-18:1 was found to be the predominant isomer (32% of total trans), while vaccenic acid was second most abundant (15% of total trans). The CLA content in longissimus lumborum was similar to that of backfat, ranging from 0.43 to 0.60% of total fatty acids and rumenic acid represented 60% of total isomers. Overall, there is still room for improvement in the saturated, mono- and polyunsaturated fatty acid composition of Canadian beef to meet general dietary guidelines for human consumption and additional targets should include reducing 10t-18:1 while increasing both rumenic and vaccenic acids.

Phenotypic correlations of fatty acid composition among intramuscular, subcutaneous and intermuscular fat tissues in concentrate-fed beef cattle of differing genotypes

[EN]In an attempt to predict intramuscular fatty acid composition using easily accessible fat depots, between-tissue correlations were studied in 75 Asturiana de los Valles bulls with different levels of muscular hypertrophy, and 25 Asturiana de la Montan˜ a bulls. Trans-18:1 in intramuscular fat was highly and positively correlated with levels in subcutaneous and intermuscular fats, while levels of total n-3 were not correlated. Predicting intramuscular fatty acid composition using easily accessible depots is thus possible for some fatty acids exhibiting high between-tissue correlations (e.g., trans-18:1) but breed and tissue specific deposition may limit this for others (e.g., n-3 fatty acids).

Extended ageing time and temperature effects on quality of sub-primal cuts of boxed beef

[EN]Most of the information indicating ageing improves tenderness has been collected on the loin and rib-eye muscles over relatively short ageing times, assuming that all muscles will react similarly. In the present study, the effect of extended ageing times on instrumental texture (56 d) and sensory characteristics (42 d) of six different beef sub-primals [striploin (SL), inside round (IR), outside round (OR), eye of round (ER), blade eye (BE) and chuck tender (CT)] was studied. The effects of two ageing temperatures (1and 58C) were also compared. In general, ageing increased tenderness (P<0.05) of SL, BE, ER and CT sub-primals, although BE shear force increased after 42 d of ageing. On the other hand, ageing had no effect on IR tenderness (P<0.05) and resulted in a decrease in tenderness of OR (P<0.05) until day 35, with a later increase after 42 d of ageing. Increasing ageing temperature (58C) had limited effect on tenderness, but ageing time and temperature increases led to lower flavour and higher off-flavour intensity (P<0.05) of the studied sub-primals. These results suggest that cutspecific maximum ageing times and rigid adherence to temperature maximums would be of benefit to optimize postslaughter processes and meat quality

Evaluation of wheat or corn dried distillers' grains with solubles on performance and carcass characteristics of feedlot steers

[EN]A study was conducted on crossbred steers (n=275; 376±924 kg) to evaluate performance and carcass quality of cattle fed wheat or corn dried distillers’ grains with solubles (DDGS). The control ration contained 86.6% rolled barley grain, 5.7% supplement and 7.7% barley silage (DM basis). The four treatments included replacement of barley grain at 20 or 40% of the diet (DM basis) with wheat or corn DDGS. Steers were slaughtered at a common end weight of 645 kg with 100 steers randomly (n=20 per treatment) selected for determination of the retail yield of sub-primal boneless boxed beef (SPBBB). Data were analyzed as a completely randomized design using pen as the experimental unit. Feeding increasing levels of wheat DDGS led to a quadratic increase in dry matter intake (DMI) (P<0.01), whereas increasing levels of corn DDGS led to a quadratic decrease in DMI (P=0.01). Average daily gain was not influenced (P=0.13) by feeding wheat or corn DDGS, but cattle fed corn DDGS exhibited a quadratic increase (P=0.01) in gain:feed. As a result, a quadratic increase (P<0.01) in calculated NEg of the diet was observed as corn DDGS levels increased. A linear decrease (P=0.04) in days on feed (169, 166 and 154 d) was noted when increasing levels of wheat DDGS (0, 20 and 40%) were fed. Dressing percentage increased in a linear fashion with wheat DDGS (P<0.01) inclusion level and in a quadratic fashion (P=0.01) as corn DDGS inclusion level increased although other carcass traits were not affected (P=0.10) by treatment. The results indicate that replacement of barley grain with corn or wheat DDGS up to 40% of the diet (DM) can lead to superior performance (improved gain:feed or reduced days on feed, respectively) with no detrimental effect on quality grade or carcass SPBBB yield.

Short communication: Rapid separation of cis9, trans11- and trans7,cis9-18:2 (CLA) isomers from ruminant tissue using a 30 m SLB-IL111 ionic column

[EN]Rumenic acid (cis9,trans11-18:2) is the main natural isomer of conjugated linoleic acid (CLA). Rumenic acid has many purported health benefits, but effects of most other CLA isomers are unknown. Typically trans7,cis9-18:2 is the second most abundant CLA isomer, but it co-elutes with rumenic acid on conventional polar gas chromatography (GC) columns, requiring complimentary analysis with silver-ion high performance liquid chromatography (Ag(+)-HPLC). Herein we report a rapid method for analyzing rumenic acid and trans7,cis9-18:2 using a 30 m ionic-liquid GC column. Optimal resolution of the two CLA isomers was at 145 degrees C and analysis of backfat from barley-fed cattle compared well with GC/Ag(+)-HPLC (y =0.978x - 0.031, r =0.985, P <0.001).

Spatial distribution of Chlorpyrifos and Endosulfan in USA coastal waters and the Great Lakes

Between 1994 and 1997, 258 tissue and 178 sediment samples were analyzed for chlorpyrifos throughout the coastal United States and the Great Lakes. Subsequently, 95 of the 1997 tissue samples were reanalyzed for endosulfan. Tissue chlorpyrifos concentrations, which exceeded the 90th percentile, were found in coastal regions known to have high agricultural use rates but also strongly correlated with sites near high population. The highest concentrations of endosulfans in contrast, were generally limited to agricultural regions of the country. Detections of chlorpyrifos at several Alaskan sites suggest an atmospheric transport mechanism. Many Great Lakes sites had chlorpyrifos tissue concentrations above the 90th percentile which decreased with increasing distance from the Corn Belt region (Iowa, Indiana, Illinois, and Wisconsin) where most agriculturally applied chlorpyrifos is used. Correlation analysis suggests that fluvial discharge is the primary transport pathway on the Atlantic and Gulf of Mexico coasts for chlorpyrifos but not necessarily for endosulfans. (PDF contains 28 pages)

Radiotherapy plus nimotuzumab or placebo in the treatment of high grade glioma patients: results from a randomized, double blind trial

Background The prognosis of patients bearing high grade glioma remains dismal. Epidermal Growth Factor Receptor (EGFR) is well validated as a primary contributor of glioma initiation and progression. Nimotuzumab is a humanized monoclonal antibody that recognizes the EGFR extracellular domain and reaches Central Nervous System tumors, in nonclinical and clinical setting. While it has similar activity when compared to other anti-EGFR antibodies, it does not induce skin toxicity or hypomagnesemia. Methods A randomized, double blind, multicentric clinical trial was conducted in high grade glioma patients (41 anaplastic astrocytoma and 29 glioblastoma multiforme) that received radiotherapy plus nimotuzumab or placebo. Treatment and placebo groups were well-balanced for the most important prognostic variables. Patients received 6 weekly doses of 200 mg nimotuzumab or placebo together with irradiation as induction therapy. Maintenance treatment was given for 1 year with subsequent doses administered every 3 weeks. The objectives of this study were to assess the comparative overall survival, progression free survival, response rate, immunogenicity and safety. Results The median cumulative dose was 3200 mg of nimotuzumab given over a median number of 16 doses. The combination of nimotuzumab and RT was well-tolerated. The most prevalent related adverse reactions included nausea, fever, tremors, anorexia and hepatic test alteration. No anti-idiotypic response was detected, confirming the antibody low immunogenicity. The mean and median survival time for subjects treated with nimotuzumab was 31.06 and 17.76 vs. 21.07 and 12.63 months for the control group. Conclusions In this randomized trial, nimotuzumab showed an excellent safety profile and significant survival benefit in combination with irradiation.

Resveratrol prevents inflammation-dependent hepatic melanoma metastasis by inhibiting the secretion and effects of interleukin-18

Background: Implantation and growth of metastatic cancer cells at distant organs is promoted by inflammation-dependent mechanisms. A hepatic melanoma metastasis model where a majority of metastases are generated via interleukin-18-dependent mechanisms was used to test whether anti-inflammatory properties of resveratrol can interfere with mechanisms of metastasis. Methods: Two experimental treatment schedules were used: 1) Mice received one daily oral dose of 1 mg/kg resveratrol after cancer cell injection and the metastasis number and volume were determined on day 12. 2) Mice received one daily oral dose of 1 mg/kg resveratrol along the 5 days prior to the injection of cancer cells and both interleukin-18 (IL-18) concentration in the hepatic blood and microvascular retention of luciferase-transfected B16M cells were determined on the 18(th) hour. In vitro, primary cultured hepatic sinusoidal endothelial cells were treated with B16M-conditioned medium to mimic their in vivo activation by tumor-derived factors and the effect of resveratrol on IL-18 secretion, on vascular cell adhesion molecule-1 (VCAM-1) expression and on tumor cell adhesion were studied. The effect of resveratrol on melanoma cell activation by IL-18 was also studied. Results: Resveratrol remarkably inhibited hepatic retention and metastatic growth of melanoma cells by 50% and 75%, respectively. The mechanism involved IL-18 blockade at three levels: First, resveratrol prevented IL-18 augmentation in the blood of melanoma cell-infiltrated livers. Second, resveratrol inhibited IL-18-dependent expression of VCAM-1 by tumor-activated hepatic sinusoidal endothelium, preventing melanoma cell adhesion to the microvasculature. Third, resveratrol inhibited adhesion-and proliferation-stimulating effects of IL-18 on metastatic melanoma cells through hydrogen peroxide-dependent nuclear factor-kappaB translocation blockade on these cells. Conclusions: These results demonstrate multiple sites for therapeutic intervention using resveratrol within the prometastatic microenvironment generated by tumor-induced hepatic IL-18, and suggest a remarkable effect of resveratrol in the prevention of inflammation-dependent melanoma metastasis in the liver.