Miniature Wulff-type generator for improving feed to fuel cells /|nT.-S. Tan. -- 260 St. Catharines, Ont. : [s. n.],

The original objective of this work was to provide a simple generator w.hich would produce hydrogen torLfuel-cell feed and which could be operated under remote or northern conditions. A secondary objective was to maximize the yield of hydrogen and carbon monoxide from available feed-stocks. A search of the patent literature has indicated that the concept of a small Wulff-type generator is essentially sound and that hydrogen may be recovered from a wide variety of hydrocarbon feed-stocks. A simple experimental set-up has been devised, patterned after ~~t originally used by R. G. Wulff for producing acetylene. This provides a supply of feed-stock, with or Without a carrier gas, which may be passed directly through a heated tube, which may contain a catalyst. A suitable procedure has been devised for analysi~ effluent gases for hydrogen, oxygen, nitrogen, methane and carbon monoxide by gas chromatography with the column packed with .Molecular .:>ieve .5 4. Athanol with air a.s carrier gas and at the same time as oxidant o was thermolyzed at temperatures in the ra~e 700-1100 C, with or Wi~lout catalyst. Methanol with or without nitrogen as a carrier gas was also cracked with • the same type of reactor refractory tube, but the temperature range was lower t down to ,300 " C when a catalyst was used. The problems of converting methane to hydrogen and carbon monoxide effiCiently, using air and/or water as oxidants were also studied.

The effects of yeast inoculation rate and acclimatization to juice on icewine fermentation /

Icewine is an intensely sweet, unique dessert wine fennented from the juice of grapes that have frozen naturally on the vine. The juice pressed from the frozen grapes is highly concentrated, ranging from a minimum of 35° Brix to approximately 42° Brix. Often Icewine fennentations are sluggish, taking months to reach the desired ethanol level, and sometimes become stuck. In 6 addition, Icewines have high levels of volatile acidity. At present, there is no routine method of yeast inoculation for fennenting Icewine. This project investigated two yeast inoculum levels, 0.2 gIL and 0.5 gIL. The fennentation kinetics of inoculating these yeast levels directly into the sterile Icewine juice or conditioning the cells to the high sugar levels using a step wise acclimatization procedure were also compared. The effect of adding GO-FERM, a yeast nutrient, was also assessed. In the sterile fennentations, yeast inoculated at 0.2 gIL stopped fennenting before the required ethanol level was achieved, producing only 7.8% (v/v) and 8.1 % (v/v) ethanol for the direct and conditioned inoculations, respectively. At 0.5 gIL, the stepwise conditioned cells fennented the most sugar, producing 12.2% (v/v) ethanol, whereas the direct inoculum produced 10.5% (v/v) ethanol. The addition of the yeast nutrient GO-FERM increased the rate of biomass accumulation, but reduced the ethanol concentration in wines fennented at 0.5 gIL. There was no significant difference in acetic acid concentration in the final wines across all treatments. Fennentations using unfiltered Icewine juice at the 0.5 gIL inoculum level were also compared to see if the effects of yeast acclimatization and micronutrient addition had the same impact on fennentation kinetics and yeast metabolite production as observed in the sterile-filtered juice fennentations. In addition, a full descriptive analysis of the finished wines was carried out to further assess the impact of yeast inoculation method on Icewine sensory quality. At 0.5 gIL, the stepwise conditioned cells fennented the most sugar, producing 11.5% (v/v) ethanol, whereas the direct inoculum produced 10.0% (v/v) ethanol. The addition of the yeast nutrient GO-FERM increased the peak viable cell numbers, but reduced the ethanol concentration in wines fennented at 0.5 gIL. There was a significant difference 7 in acetic acid concentration in the final wines across all treatments and all treatments affected the sensory profiles of the final wines. Wines produced by direct inoculation were described by grape and raisin aromas and butter flavour. The addition of GO-FERM to the direct inoculation treatment shifted the aroma/flavour profiles to more orange flavour and aroma, and a sweet taste profile. StepWise acclimatizing the cells resulted in wines described more by peach and terpene aroma. The addition of GO-FERM shifted the profile to pineapple and alcohol aromas as well as alcohol flavour. Overall, these results indicate that the addition of GO-FERM and yeast acclimatization shortened the length of fermentation and impacted the sensory profiles of the resultant wines.

Transfection of newt blastema mesenchyme using the techniques of lipofection and direct gene transfer

The regenerating amphibian limb provides a useful system for studying genes involved in the establishment of positional information. While a number of candidate genes that may playa role in pattern formation have been identified, their function in vivo is unknown in this system. To better ascertain the role of these genes, it would be useful to be able to alter their normal patterns of expression in vivo and to assess the effects of this misexpression on limb pattern. In order to achieve this, a method of introducing a plasmid containing the eDNA of a gene of interest into a newt blastema (a growth zone of mesenchymal progenitor cells) is needed. Unfortunately, most commonly used transfection techniques cannot be used with newt blastema cells. In this study, I have used the techniques of lipofection and direct gene transfer to introduce plasmid DNA containing reporter genes into the cells of a regenerating newt limb. The technique of lipofection was most effective when the blastema cells were transfected in vitro. The optimal ratio for transfection was shown to be 1:3 DNA:Lipofectin (W/w) , and an increase in the amount of DNA present in the mixture (1:3 ratio maintained) resulted in a corresponding increase in gene expression. The technique of direct gene transfer was used to transfect newt blastema cells with and without prior complex formation with Lipofectin. Injection of plasmid DNA alone provided the most 3 promising results. It was possible to introduce plasmid DNA containing the reporter gene ~-galactosidase and achieve significant gene expression in cells associated with the injection site. In the future, it would be interesting to use this technique to inject plasmid DNA containing a gene which may have a role in pattern formation into specific areas of the newt blastema and to analyze the resulting limb pattern that emerges.

Biological effects of resveratrol on skeletal muscle cells

Resveratrol, a polyphenol found in red wine, has been reported to have antithrombotic, antiatherogenic, and anticancer properties both in vitro and III VIVO. However, possible antidiabetic properties of resveratrol have not been examined. The objective of this study was to investigate the direct effects of resveratrol on basal and insulin-stimulated glucose uptake and to elucidate its mechanism of action in skeletal muscle cells. In addition, the effects of resveratrol on basal and insulin- stimulated amino acid transport and mitogenesis were also examined. Fully differentiated L6 rat skeletal muscle cells were incubated with resveratrol concentrations ranging from 1 to 250 IlM for 15 to 120 min. Maximum stimulation, 201 ± 8.90% of untreated control, (p<0.001), of2eH] deoxy- D- glucose (2DG) uptake was seen with 100 IlM resveratrol after 120 min. Acute, 30 min, exposure of the cells to 100 nM insulin stimulated 2DG uptake to 226 ± 12.52% of untreated control (p<0.001). This appears to be a specific property of resveratrol that is not shared by structurally similar antioxidants such as quercetin and rutin, both of which did not have any stimulatory effect. Resveratrol increased the response of the cells to submaximal insulin concentrations but did not alter the maximum insulin response. Resveratrol action did not require insulin and was not blocked by the protein synthesis inhibitor cycloheximide. L Y294002 and wortmannin, inhibitors of PI3K, abolished both insulin and resveratrolstimulated glucose uptake while phosphorylation of AktlPKB, ERK1I2, JNK1I2, and p38 MAPK were not increased by resveratrol. Resveratrol did not stimulate GLUT4 transporter translocation in GLUT4cmyc overexpressing cells, in contrast to the significant translocation observed with insulin. Furthermore, resveratrol- stimulated glucose transport was not blocked by the presence of the protein kinase C (PKC) inhibitors BIMI and G06983. Despite that, resveratrol- induced glucose transport required an intact actin network, similar to insulin. In contrast to the stimulatory effect seen with resveratrol for glucose transport, e4C]methylaminoisobutyric acid (MeAIB) transport was inhibited. Significant reduction of MeAIB uptake was seen only with 100uM resveratrol (74.2 ± 6.55% of untreated control, p<0.05), which appeared to be maximum. In parallel experiments, insulin (100 nM, 30 min) increased MeAIB transport by 147 ± 5.77% (p<0.00l) compared to untreated control. In addition, resveratrol (100 JlM, 120 min) completely abolished insulin- stimulated amino acid transport (103 ± 7.35% of untreated control,p>0.05). Resveratrol also inhibited cell proliferation in L6 myoblasts with maximal inhibition of eH]thymidine incorporation observed with resveratrol at 50 J.LM after 24 hours (8 ± 1.59% of untreated control, pcells by a mechanism that is in4ependent of insulin and protein synthesis. Resveratrol- stimulated glucose uptake may be PI3K and actin cytoskeleton- dependent and independent of AktIPKB, PKC, ERK1I2, JNK1I2, p38 MAPK, and GLUT4 translocation. However, unlike glucose transport, resveratrol inhibits both basal and insulin- stimulated amino acid transport and mitogenesis.