Thermally-dried immobilized kefir on casein as starter culture in dried whey cheese production.

The aim of the present study was to evaluate the use of thermally-dried immobilized kefir on casein as a starter culture for protein-enriched dried whey cheese. For comparison reasons, dried whey cheese with thermally-dried free kefir culture and with no starter culture were also produced. The effect of the nature of the culture, the ripening temperature and the ripening process on quality characteristics of the whey cheese was studied. The association of microbial groups during cheese maturation suggested repression of spoilage and protection from pathogens due to the thermally-dried kefir, as counts of coliforms, enterobacteria and staphylococci were significantly reduced in cheeses produced using thermally-dried kefir starter cultures. The effect of the starter culture on production of volatile compounds responsible for cheese flavor was also studied using the SPME GC/MS technique. Thermally-dried immobilized kefir starter culture resulted in an improved profile of aroma-related compounds. The preliminary sensory evaluation ascertained the soft, fine taste and the overall improved quality of cheese produced with the thermally-dried immobilized kefir. The potential of protein-based thermally-dried starter cultures in dairy products is finally highlighted and assessed.

Probiotic cheese production using Lactobacillus casei cells immobilized on fruit pieces.

Lactobacillus casei cells were immobilized on fruit (apple and pear) pieces and the immobilized biocatalysts were used separately as adjuncts in probiotic cheese making. In parallel, cheese with free L. casei cells and cheese only from renneted milk were prepared. The produced cheeses were ripened at 4 to 6 degrees C and the effect of salting and ripening time on lactose, lactic acid, ethanol concentration, pH, and lactic acid bacteria viable counts were investigated. Fat, protein, and moisture contents were in the range of usual levels of commercial cheeses. Reactivation in whey of L. casei cells immobilized on fruit pieces after 7 mo of ripening showed a higher rate of pH decrease and lower final pH value compared with reactivation of samples withdrawn from the remaining mass of the cheese without fruit pieces, from cheese with free L. casei, and rennet cheese. Preliminary sensory evaluation revealed the fruity taste of the cheeses containing immobilized L. casei cells on fruit pieces. Commercial Feta cheese was characterized by a more sour taste, whereas no significant differences concerning cheese flavor were reported by the panel between cheese containing free L. casei and rennet cheese. Salted cheeses scored similar values to commercial Feta cheese, whereas unsalted cheese scores were significantly lower, but still acceptable to the sensory panelists.

High-temperature wine making using the thermotolerant yeast strain Kluyveromyces marxianus IMB3.

Kluyveromyces marxianus IMB3 yeast cells were immobilized on delignified cellulosic material, apple, and quince separately. Both immobilized and free cells were used in high-temperature wine making, and their fermented grape must contained 3 to 4% alcohol. Semisweet wines were produced by the addition of potable alcohol to the fermented must. Preliminary sensory evaluation of the produced semisweet wines showed good flavor and aroma. The final product contained extremely low levels of higher and amyl alcohols while ethyl acetate was at levels usually present in wines. The ferment produced may be blended with other products to improve their quality.

Disruption of the small-intestine mucosal barrier after intestinal occlusion: a study with light and electron microscopy.

It is known that the gut may serve as a reservoir for various microorganisms, which under specific circumstances may intrude into the systemic circulation, causing systemic infections. The aim of the present study was to estimate the "critical time" of disruption of the small-intestine mucosal barrier in conditions of experimentally induced intestinal occlusion, based on the histopathological alterations observed under light and electron microscopy. Forty rabbits underwent small-intestine obstruction through ligation with a nonabsorbable suture. Blood cultures from portal vein and inferior vena cava, as well as cultures from the peritoneal fluid, a hepatic fragment, and a mesenteric lymph node, were obtained before the ligation (0 h). The same cultures were repeated at 4 and 8 h (group A, 20 rabbits) and at 6 and 12 h after the ligation (group B, 20 rabbits). Small-intestine specimens proximal to the occlusion were taken for examination under the optic and electronic microscope in the same time intervals. Five of 20 rabbits of group A died within 4 h and 6 of 20 rabbits of group B died within 6 h after the operation. All anaerobic cultures were negative. All aerobic cultures that became positive developed Escherichia coli colonies. Intestinal epithelium of dead animals was transformed to cuboid with destruction of goblet cells and alteration in secretion of acid polysaccharides. The mucosal appearance of all rabbits that survived 12 hours after ligation was the same. The disruption of the mucosal barrier begins 4 h after complete intestinal occlusion. At 12 h after complete intestinal occlusion, the disruption is total with different degrees of severity.

Grape and apple wines volatile fermentation products and possible relation to spoilage.

The main volatile by-products of the alcoholic fermentation of grape wine, cider and apple pulp wine were investigated to determine if any correlated with spoilage resistance in the latter two. Spoilage was visually detected after seven days in low-alcohol grape wine in comparison to 11 and 16 days in cider and apple pulp wine, respectively. Acetaldehyde, ethyl acetate, methanol, propanol, isobutanol and amyl alcohols were the main fermentation by-products detected in all three wines. There were highest concentrations of acetaldehyde, ethyl acetate, methanol and propanol in grape wine and, therefore, these by-products could not be implicated in spoilage resistance in apple wines. Increased concentrations of isobutanol and amyl alcohols, however, in cider and apple pulp wine in comparison to grape wine might have been the reason for spoilage resistance in the apple wines.

Effect of freeze-dried immobilized cells on delignified cellulosic material in low-temperature and ambient-temperature wine making.

In this article, we report on wine making by freeze-dried immobilized cells on delignified cellulosic material for ambient and low temperatures. Biocatalyst supported by freeze-dried delignified cellulosic (FDC) material recovered after the first repeated-batch fermentations the fermentation efficiency and startup, which become about equal to those of biocatalyst supported by wet delignified cellulosic material. The FDC biocatalyst was suitable for wine making at low temperatures (5-15 degrees C), and produced wine of 12% alcoholic degree, with the main volatiles contained in the wine and reduced by a decrease in temperature. The fermentation efficiency was not affected by total acidity of must, while an increase in initial Be density improved percentages of higher alcohols and ethyl acetate. The quality of the wine was validated by a preliminary taste test to be in the range of acceptable to excellent.

Industrial scale pilot plant operation by Kefir yeast cells immobilized on delignified cellulosic material for whey treatment.

Kefir yeast cells immobilized on Delignified Cellulosic Materials (DCM) is proposed as a suitable form of biocatalyst for whey treatment. A bioreactor of 11.000L was used in all experiments. 5 repeated batch fermentations were taken place with whey volumes of 3000L and 5000L. Fermentation times achieved were below 15 h and final ethanol concentration was of the level of 2%v/v. Residual sugar was low while conversion rate reached 95%. In a second frame of experiments the immobilized cells bioreactor was operated on a feed-batch process, beginning from 2000L of whey and adding 2000L in every feed. In a third frame of experiments, whey fermentation was performed on a continuous operation for a period of 13 days. The system retained its operational stability, exhibiting relatively high ethanol yield and ethanol productivity. All experiments indicated that the biocatalyst of kefir yeast cells immobilized on delignified cellulosic materials has a carry through property on whey fermentation on industrial scale.

Whey liquid waste of the dairy industry as raw material for potable alcohol production by kefir granules.

Kefir granular biomass was used in the fermentation of sweet whey and proved to be more effective compared to single-cell biomass of kefir yeast. The operational stability of the biocatalyst was assessed by carrying out 20 repeated batch fermentations. Levels of ethanol productivity reached 2.57 g L(-1) h(-1)), whereas the yield was 0.45 g/g. The fermentation time was only 8 h. Mixtures of sweet whey with molasses were fermented at initial densities ranging from 4.2 to 10.2 degrees Be and resulted in ethanol yield factors between 0.36 and 0.48 g of ethanol/g of utilized sugar. Lower degrees Be values led to an increase of percentages of ethyl acetate on total volatiles determined and a reduction of amyl alcohols. The addition of 1% black raisin extract to whey appears to promote whey fermentation, whereas the same was not observed in the case of white sultana extract addition. It was finally established that it is preferable to ferment mixtures of whey-molasses by adding molasses in whey after the completion of whey fermentation.

Low-temperature brewing using yeast immobilized on dried figs.

Dried figs, following exhaustive extraction of their residual sugars with water, were used for immobilization of Saccharomyces cerevisiae AXAZ-1. The immobilized biocatalyst was used in repeated batch fermentations of glucose at 30 degrees C, where significant reduction of the fermentation time was observed, falling from 65 h in the first batch to 7 h after the sixth batch. Repeated fermentations of wort at room and low temperatures resulted in fermentation times that fell from 26 to 20 h and from 27 to 24 days at 18 and 3 degrees C, respectively. Ethanol and beer productivities were high, showing suitability of the biocatalyst for low-temperature brewing. Diacetyl concentrations were low (0.3-0.5 mg/L), and polyphenols were lower than in commercial products and decreased as the fermentation temperature was decreased (126-50 mg/L). Ethyl acetate concentrations increased from 53 to 88 mg/L as the temperature was decreased, while the concentration of amyl alcohols at 3 degrees C (58 mg/L) was lower than half of that at 18 degrees C (125 mg/L). The beers produced at the end of the main fermentation had a fine clarity and a special fruity figlike aroma and taste, distinct from commercial products and more intense than beers produced by cells immobilized on other food-grade supports (gluten pellets or delignified cellulosic materials). GC-MS analysis did not show significant differences in the qualitative composition of the aroma compounds of the beers produced by immobilized and free cells.

High-temperature alcoholic fermentation of whey using Kluyveromyces marxianus IMB3 yeast immobilized on delignified cellulosic material.

A novel system for high-temperature alcoholic fermentation of whey is described. This system consists of Kluyveromyces marxianus yeast immobilized on delignified cellulosic material (DCM). The effect of pH, initial lactose concentration and temperature on the fermentation of a synthetic medium containing lactose was studied. Batch fermentations of whey were also carried out and the formation of volatile by-products was examined. The concentrations of higher alcohols were found to be in very low levels leading to a product of improved quality. The fermented whey had an improved characteristic aroma compared to unfermented whey. The possibility to use fermented whey as raw material for the production of a novel, low alcohol content drink was also investigated.

Continuous whey fermentation using kefir yeast immobilized on delignified cellulosic material.

Delignified cellulosic-supported biocatalyst, prepared by immobilization of kefir yeast on delignified cellulosic material (DCM), was found to be suitable for continuous, modified whey fermentation. The modified whey contained 1% raisin extract and molasses. Ethanol productivities ranged from 3.6 to 8.3 g L(-1)day(-1), whereas parameters such as ethanol concentration, residual sugars, and daily fermented whey productivity were acceptable for the production of potable alcohol and alcoholic drinks in industrial fermentations. The continuous fermentation bioreactor was operated for 39 days, stored for 18 days at 4 degrees C, and operated again for another 15 days without any diminution of the ethanol productivity. The concentrations of higher alcohols (propanol-1, isobutyl alcohol, and amyl alcohols) were low. The main volatile byproducts formed in the continuous process were similar to those observed in alcoholic beverages, and the fermented whey had a good aroma. The concentrations of higher alcohols were very low when compared to that of ethyl acetate, therefore resulting in a quality product. The possibility of using such a process for the production of potable alcohol or a novel, low-alcohol content drink is proposed.

The effect of local application of mitomycin-C on the development of capsule around silicone implants in the breast: an experimental study in mice.

In this experimental study on mice we try to prove that capsule formation around breast implants is considerably diminished after topical application of Mitomycin-C (MMC). MMC solution is applied under the breast tissue or pectoralis major muscle, in the pocket of the silicone implant and just before placing it. This is a morphological study of the connective tissue formed around the implant, under light and electron microscope. Our results suggest that it is worthwhile working on a clinical study trying to prevent the most common complication of breast implants, contracture of a capsule.

Wine production using yeast immobilized on apple pieces at low and room temperatures.

A biocatalyst was prepared by immobilization of Saccharomyces cerevisiae strain AXAZ-1 on apple pieces. It was examined by electron microscope and studied during the fermentation of grape must for batch wine-making. The immobilized yeast showed an important operational stability without any decrease of its activity even at low temperatures (1-12 degrees C). Specifically, at 6 degrees C the biocatalyst favored wine production within 8 days, which is less time than is required for the natural fermentation of grape must. At 1 degrees C wine production was effected in 1 month. This speeding up of the fermentation could be accepted and adopted by the industry for scaling up the wine-making process. Total and volatile acidities were similar to those found in dry wines. The concentrations of higher alcohols (propanol-1 and isobutyl alcohol) were low. The presence of amyl alcohols proved to be temperature dependent and decreased with the temperature decrease. The values of ethyl acetate concentrations were relatively high, up to 154 mg/L. This probably contributes to the fruity aroma and excellent taste of the produced wines. There was no indication of vinegar odor in the product given that the volatile acidity was <0.47 g of acetic acid/L. From the GC-MS analysis it was concluded that cell immobilization did not create serious changes in the product (wine) with regard to the qualitative composition of the aroma compounds.

Effect of carbohydrate substrate on fermentation by kefir yeast supported on delignified cellulosic materials.

The suitability of delignified cellulosic (DC) material supported kefir yeast to ferment raw materials that contain various single carbohydrates, for the production of potable alcohol and alcoholic drinks, is examined in this investigation. Results are reported of fermentations carried out with sucrose, fructose, and glucose in synthetic media. Repeated batch fermentations at various initial sugar concentrations of sucrose, fructose, and glucose were performed at 30 degrees C in the presence of the aforementioned biocatalyst. The results clearly show feasible yields in the range of 0.38-0.41 g/g, alcohol concentrations of 7.6-8.2% v/v, fermentation time of 90-115 h, and conversion of 92-96%. DC material supported kefir fermented 11-fold more rapidly than free cells and 9-fold more rapidly in comparison to kissiris supported kefir. The main volatile byproducts such as amyl alcohols (mixture of 2-methyl-1-butanol and 3-methyl-1-butanol), ethanal, and ethyl acetate were formed in all sugar fermentation products. The formation of 65-110 ppm of ethyl acetate is as high and even higher than that obtained with traditional wine yeasts. The increase of the initial concentration of sugar in the fermentation media resulted in an increase in contents of volatiles. The fine aroma that was obtained in the product of fructose could be attributed to the high percentage of ethyl acetate on total volatiles. The efficiency of DC material supported kefir was the same for the fermentations of individual sugars or a mixture of fructose, sucrose, and glucose. When whey with raisin extracts was fermented, lower yields were obtained but the aroma of the product was even better.

Low-temperature brewing by freeze-dried immobilized cells on gluten pellets.

A biocatalyst, prepared by the immobilization of a cryotolerant strain of Saccharomyces cerevisiae on gluten pellets, was freeze-dried without any protecting medium and used for repeated batch fermentations of wort for each of the temperatures 15, 10, 5, and 0 degrees C. The fermentation time for freeze-dried immobilized cells was about 2-fold that of the corresponding time for wet immobilized cells on gluten pellets, and lower than the corresponding time for freeze-dried free cells, especially at 5 and 0 degrees C. Beers produced by freeze-dried immobilized cells contained alcohol levels in the range of 5.0-5.5% v/v, diacetyl concentrations lower than 0.5 mg/L, polyphenol concentrations lower than 145.5 mg/L, and free cell concentrations lower than 3 g/L. As a result, they had a very good clarity after the end of primary fermentation. The amounts of amyl alcohols were lower than 129.1 mg/L and reduced as the temperature was decreased. Ethyl acetate concentrations were found in the range of 22.1-29.2 mg/L, giving a very good aroma and taste in the produced beers.

Continuous wine making by gamma-alumina-supported biocatalyst: quality of the wine and distillates.

The main objective of the present work was the removal of aluminum from wines produced by gamma-alumina-supported yeast cells. Reagents such as Na2CO3, NH4OH, albumin, and Ca(OH)2 were used. Calcium in the presence of albumin was effective, whereas other reagents were not so effective. Because of the improved aroma and taste of distillates produced by gamma-alumina-supported biocatalyst, volatile byproducts of distillates were analyzed. They were also assessed by sensory tests. Methanol, acetaldehyde, ethyl acetate, propanol-1, isobutyl alcohol, and amyl alcohols were determined in distillates. It was noted that the amounts of higher alcohols and amyl alcohols decreased as the temperature of fermentation dropped, leading to a product of improved quality and reduced toxicity.

Delignified cellulosic material supported biocatalyst as freeze-dried product in alcoholic fermentation.

Freeze-dried delignified cellulosic (DC) material supported biocatalyst is proposed as a suitable form of biocatalyst to be preserved. The alcoholic fermentation of glucose using freeze-dried immobilized cells is reported. Freeze-dried immobilized baker's yeast cells on DC material do not need any protective medium during freeze-drying. The effect of initial glucose concentration and temperature on the alcoholic fermentation kinetic parameters is reported in the present study. It was found that the freeze-dried immobilized cells ferment more quickly than free freeze-dried cells and have a lower fermentation rate as compared with wet immobilized cells. However, repeated batch fermentations showed freeze-dried immobilized cells to ferment at about the same fermentation rate as wet immobilized cells. The results indicate that the freeze-dried immobilized cells must be further studied to establish a process for the preservation of immobilized cells.

Low-temperature alcoholic fermentation by delignified cellulosic material supported cells of kefir yeast.

A novel system for low-temperature alcoholic fermentation of glucose is described. This system consists of kefir yeast immobilized on delignified cellulosic materials. Batch fermentations were carried out at various pH values, and the effect of temperature on kinetic parameters, in the range of 5-30 degrees C, was examined. At pH 4.7 the shortest fermentation time was obtained. The formation of volatiles indicates that the concentration of amyl alcohols (total content of 2-methylbutanol-1 and 3-methylbutanol-1) is reduced as the temperature becomes lower. Propanol-1 and isobutyl alcohol formation drops significantly below 15 degrees C. The percentage of ethyl acetate increases as the temperature is diminished. At 5 degrees C the content of total volatiles in the product was only 38% of the volatiles formed during fermentation at 30 degrees C.

Lung carcinoma imaging using a synthetic laminin derivative radioiodinated peptide YIGSR.

UNLABELLED: The synthetic laminin pentapeptide tyrosyl-isoleucyl-glycyl-seryl-arginine (YIGSR) binds to a metastasis associated high-affinity laminin receptor. The aim of this study was to investigate if the radiolabeled peptide can be considered as a basis for a potential tumor-imaging radiopharmaceutical. METHODS: Iodine-131-labeled YIGSR was injected in mice inoculated with Lewis Lung carcinoma, as well as in normal controls. The experimental animals were imaged on a gamma camera 10 hr after peptide administration. The same peptide was also labeled with 125I and administered to tumor-bearing and normal mice. At various time-points after peptide administration, the experimental animals were killed, and the radioactivity in the tumor, lung, liver and spleen was measured. Microscopic autoradiography was performed in histological sections of the same tissues. RESULTS: The tumor and the spleen of tumor-bearing animals were imaged on a gamma camera. No significant blood-pool background was detected. No other organ except urinary bladder and thyroid was imaged in normal animals. The peptide was retained on tumor and spleen of tumor-bearing animals. Twenty-four hours after peptide administration, the tumor, lung, liver and spleen of animals with tumors contained significantly more radioactivity than the same tissues of equally treated normal controls. The radiolabeled peptide YIGSR was detected by microscopic autoradiography on the surface of certain tumor cells, but not on the surface of any normal cell. CONCLUSION: Although extensive research is still required, the peptide YIGSR can be considered as a basis for the development of a receptor specific radiopharmaceutical useful for the in vivo estimation of the metastatic potential of tumors. This radiopharmaceutical may be helpful in staging and prognostic-related decisions on cancer treatment.

Continuous wine making by delignified cellulosic materials supported biocatalyst. An attractive process for industrial applications.

The continuous making of wine by a delignified cellulosic (DC) material-supported biocatalyst is reported. It was prepared by immobilizing the alcohol resistant strain AXAZ-1 on DC material. The product was found suitable for the continuous process in industrial applications. The operational stability was maintained for 2 mo with monitoring the ethanol concentration, wine, and alcohol productivities as well as the stability of oBe density at the outlet. Wine productivity was three- to sixfold higher than obtained by natural fermentation, alcohol concentrations of the wine was in the range of 9.3-11.2% v/v and low volatile acidities of 0.15-0.36 g acetic acid/L were obtained. The effect of total acidity and flow rate of must were also examined. To demonstrate that the operational stability of the bioreactor is due to DC material that promotes the fermentation, and it takes place at even higher ethanol levels, an analogous system of kissiris supported biocatalyst was studied. Likewise, the tolerance in the alcohol concentration, as compared with free cells, were studied by their stability of the activity in the repeated batch fermentation of must.