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Biomedical subjects

R Lanciotti

Publications and source records attributed to R Lanciotti.

At least 19 recordsLinked to original sources

Generation of aroma compounds in sourdough: effects of stress exposure and lactobacilli-yeasts interactions.

The effects of the interaction between Saccharomyces cerevisiae LBS and Lactobacillus sanfranciscensis LSCE1 and of their responses to acid, oxidative or osmotic stress on alcohol and aroma production were assessed. The exposure of S. cerevisiae LBS and L. sanfranciscensis LSCE1 cells to oxidative, acid or osmotic sub-lethal stress gave rise to a common or specific responses. Gamma-decalactone, 2(5H)-furanones and aldehydes were overproduced by LAB following oxidative stress. The acid stress induced both in yeasts and LAB, as well as in their co-cultures, a relevant accumulation of isovaleric and acetic acids and higher alcohols. A cross-exposure of yeasts and LAB to their preconditioned media, generated in S. cerevisiae a release of esters including esters of long-chain unsaturated fatty acids coming from membrane phospholipids. These esters were excreted also by yeasts following a pressure stress.

Alcohols↗

Characterization of yeasts involved in the ripening of Pecorino Crotonese cheese.

The aims of this work were to identify and characterize for some important technological properties the yeast species present throughout the ripening process of Pecorino Crotonese, a traditional cheese produced in a well defined area of Southern Italy. In particular, the strain technological properties considered include fermentation/assimilation of galactose and lactose, assimilation of lactate and citrate in the presence of different NaCl concentrations, hydrolysis of butter fat, skim milk, gelatine and casein, production of brown pigments in cheese agar and ability to produce biogenic amines. High yeast levels were recorded in cheese samples already after 5 h of brining (about 5 log cfu/g) and these concentration remained constant during ripening. The yeast isolates belonged to restrict number of yeast species. While Kluyveromyces lactis and Saccharomyces cerevisiae were isolated prevalently in the first stages of Pecorino Crotonese production, Yarrowia lipolytica and Debaryomyces hansenii dominated during the later stages of maturation. Otherwise, the latter two were very NaCl resistant species. In fact, D. hansenii strains conserved the ability to assimilate lactose and galactose in the presence of 10% NaCl, while almost all the strains of Y. lipolytica isolated assimilated citrate and lactate up to 7.5% NaCl. Y. lipolytica isolates evidenced also the highest proteolytic and lipolytic activities and the capability to catabolize tyrosine producing brown pigment. In addition they resulted in the highest aminobiogenic potential decarboxylating ornithine, phenylalanine, tyrosine and lysine. However, they were not able to produce histamine, biogenic amine produced by three strains of D. hansenii.

Carbohydrate Metabolism↗

Use of Yarrowia lipolytica strains for the treatment of olive mill wastewater.

The principal aim of this research was to evaluate the ability of different Yarrowia lipolytica strains, having different origin, to grow in olive mill wastewater (OMW) and reduce its COD level. All the strains were able to grow in undiluted OMW; the comparison between the data obtained in a semi-synthetic medium and in OMW suggests that lipases with different specificity can be produced in relation to the medium composition. Under the adopted conditions, the reduction of the OMW COD values varied from 1.47% and 41.22% of the initial value. Some strains determined a significant reduction of polyphenol content, while other ones caused its apparent increase. Moreover, some Y. lipolytica strains, isolated from chilled foods, produced the highest citric acid concentrations. These results evidenced that some Y. lipolytica strains are good candidates for the reduction of the pollution potential of OMW and for the production of enzymes and metabolites such as lipase and citric acid.

Citric Acid↗

Interactions between high pressure homogenization and antimicrobial activity of lysozyme and lactoperoxidase.

It was the objective of this work to evaluate the effect of high pressure homogenization on the activity of antimicrobial enzymes such as lysozyme and lactoperoxidase against a selected group of Gram positive and Gram negative species inoculated in skim milk. Lactobacillus helveticus, Lactobacillus plantarum and Listeria monocytogenes were the most pressure resistant species while Bacillus subtilis, Pseudomonas putida, Salmonella typhimurium, Staphylococcus aureus, Proteus vulgaris and Salmonella enteritidis were found to be very sensitive to the hyperbaric treatment. The enzyme addition enhanced the instantaneous pressure efficacy on almost all the considered species as indicated by their instantaneous viability loss following the treatment. Moreover, the combination of the enzyme and high pressure homogenization significantly affected the recovery and growth dynamics of several of the considered species. Although L. monocytogenes was slightly sensitive to pressure, the combination of the two stress factors induced a significant viability loss within 3 h and an extension of lag phases in skim milk during incubation at 37 degrees C. The hypothesis formulated in this work is that the interaction of high pressure homogenization and lysozyme or lactoperoxidase is associated to conformational modifications of the two proteins with a consequent enhancement of their activity. This hypothesis is supported by the experimental results also regarding the increased antimicrobial activity against L. plantarum of the previously pressurised lysozyme with respect to that of the native enzyme.

Animals↗

Proteolytic, lipolytic and molecular characterisation of Yarrowia lipolytica isolated from cheese.

This work studied the qualitative and quantitative proteolytic and lipolytic activities of Yarrowia lipolytica strains isolated from two cheese types. Randomly amplified polymorphic DNA-PCR (RAPD-PCR) analysis was used to compare the cheese strains of Y. lipolytica with strains isolated from other food products and with the type strain of the species in order to investigate the genetic diversity and occurrence of specific environmental groups. Diversity of proteolytic and especially lipolytic activity within Y. lipolytica strains isolated from dairy products was observed. In particular, the degree of specificity for saturated or unsaturated fatty acids as well as for even- or odd-numbered carbon free fatty acids (FFAs) varied among the strains. The RAPD-PCR profiles showed low genetic relatedness between many of the food isolates and the type strain of the species. Such genetic variability needs to be further evaluated. Most of the Y. lipolytica strains appeared to be specific to the particular environment from which they were isolated. However, phenotypic characteristics having technological importance in dairy products and, particularly, lipolytic activities did not correspond to the genetic differences observed by RAPD-PCR analysis.

Cheese↗

Variability of the lipolytic activity in Yarrowia lipolytica and its dependence on environmental conditions.

This work was aimed to the evaluation of the variability of lipolytic activity in Yarrowia lipolytica strains, as well as to asses for a selected strain, the response to the changes of physico-chemical variables (such as pH, NaCl and lipid content), in order to obtain predictive models describing their effects on the lipolysis pattern. The strains tested, having different environmental origin, showed different patterns of the free fatty acids (FFA) released. The clustering of the free fatty acids profiles evidenced that the unweighted average distance within the strains of the same species did not exceeded 30%. However, the lipolytic activity of some strains generated FFA profiles that differentiated from the majority of the strains considered. Also, when a single strain was inoculated in model systems in which pH, NaCl and milk fat were modulated according to a Central Composite Design (CCD), chemico-physical characteristics of the system led to marked variations in the lipolytic activity with consequent changes in individual fatty acids released. In most cases, when the same Y. lipolytica strain was used, under the experimental conditions adopted, the modulation of the lactic acid, NaCl and lipid content did not generate differences in the fatty acid release exceeding 20-21%. However, some combinations of factors remarkably affected lipase expression or activity, and generated differences in the fatty acid released higher than those observed among different strains of the same species.

Animals↗

Occurrence and characterization of yeasts isolated from milks and dairy products of Apulia region.

One hundred and five strains of yeasts isolated from milk of different animal origin and from typical Apulian cheeses were studied to identify and characterize yeast strains for further selection as starter cultures for cheese production. The most prevalent isolates belonged to the species Trichosporon cutaneum (15.24%), Candida catenulata (10.48%) and Yarrowia lipolytica (8.57%). In order to evaluate the potential use as starter cultures, the occurrence of some selected properties, such as fermentation of glucose and lactose, assimilation of lactic acid, citric acid and lactose, growth at 4 degrees C and production of lipolytic and proteolytic enzymes in the strains belonging to the most frequent species was tested. C. catenulata and C. zeylanoides were positive for assimilation of lactic and citric acids and showed psychrotrophic aptitude. T. cutaneum was positive for all properties tested except for glucose and lactose fermentations. Y. lipolytica was endowed with remarkable lipolytic activity also at 4 degrees C and was positive for assimilation of lactic and citric acids, growth at 4 degrees C and proteolytic activity.

Animals↗

Vanadate-induced cell growth regulation and the role of reactive oxygen species.

While vanadium compounds are known as potent toxicants as well as carcinogens, the mechanisms of their toxic and carcinogenic actions remain to be investigated. It is believed that an improper cell growth regulation leads to cancer development. The present study examines the effects of vanadate on cell cycle control and involvement of reactive oxygen species (ROS) in these vanadate-mediated responses in a human lung epithelial cell line, A549. Under vanadate stimulation, A549 cells generated hydroxyl radical (*OH), as determined by electron spin resonance (ESR), and hydrogen peroxide (H2O2) and superoxide anion (O2*-), as detected by flow cytometry using specific dyes. The mechanism of ROS generation involved the reduction of molecular oxygen to O2*- by both a flavoenzyme-containing NADPH complex and the mitochondria electron transport chain. The O2*- in turn generated H2O2, which reacted with vanadium(IV) to generate *OH radical through a Fenton-type reaction (V(IV) + H2O2 --> V(V) +*OH + OH-). The ROS generated by vanadate induced G2/M phase arrest in a time- and dose-dependent manner as determined by measuring DNA content. Vanadate also increased p21 and Chk1 levels and reduced Cdc25C expression, leading to phosphorylation of Cdc2 and a slight increase in cyclin B1 expression as analyzed by Western blot. Catalase, a specific antioxidant for H2O2, decreased vanadate-induced expression of p21 and Chk1, reduced phosphorylation of Cdc2Tyr15, and decreased cyclin B1 levels. Superoxide dismutase, a scavenger of O2*-, or sodium formate, an inhibitor of *OH, had no significant effects. The results obtained from the present study demonstrate that among ROS, H2O2 is the species responsible for vanadate-induced G2/M phase arrest. Several regulatory pathways are involved: (1) activation of p21, (2) an increase of Chk1 expression and inhibition of Cdc25C, which results in phosphorylation of Cdc2 and possible inactivation of cyclin B1/Cdc2 complex.

Anions↗

Purification and characterization of an extracellular proline iminopeptidase from Corynebacterium variabilis NCDO 2101.

AIMS: To screen the extracellular proteolytic and lipolytic activities of Corynebacterium variabilis NCDO 2101 and to purify and characterize a proline iminopeptidase enzyme in order to investigate the role of the major component of the smear of bacterial surface-ripened cheeses. METHODS AND RESULTS: Four chromatographic steps were used to purify the enzyme and a three-factor, five-level Central Composite Design was used to study the interactive effects of cheese-related values of pH, NaCl and temperature. The proline iminopeptidase showed some biochemical properties different from the same enzyme purified from lactic acid bacteria and other smear bacteria. It tolerated NaCl concentrations up to 7.5% and was sensitive to low values of pH especially when they were combined with low temperature. CONCLUSION: The proline iminopeptidase of C. variabilis NCDO 2101 may have a role in proteolysis during ripening of smear surface-ripened cheeses. SIGNIFICANCE AND IMPACT OF THE STUDY: The findings of this work contribute to the knowledge of the enzymology of smear bacteria in order to improve the ripening of bacterial surface-ripened cheeses.

Aminopeptidases↗

Effect of trans-2-hexenal on the growth of Aspergillus flavus in relation to its concentration, temperature and water activity.

AIMS: The antifungal activity of trans-2-hexenal on Aspergillus flavus in a model system in relation to its concentration, incubation temperature and aw was assessed. METHODS AND RESULTS: A model describing the antifungal activity of the aldehyde in relation to these variables was obtained. CONCLUSION: According to this model, the inhibition of A. flavus was weakly dependent on the incubation temperature (at least within the range of values considered) and strongly affected by the trans-2-hexenal concentration and aw, which showed a remarkable synergistic effect. SIGNIFICANCE AND IMPACT OF THE STUDY: Trans-2-hexenal proved to be a molecule with remarkable antimicrobial properties, even when added in closed systems at low concentration.

Aldehydes↗

Suitability of log-linear models to evaluate the microbiological quality of baby clams (Chamelea gallina L.) harvested in the Adriatic Sea.

The presence of fecal coliforms or Escherichia coli in baby clams (Chamelea gallina L.) is considered an indicator related to their safety because they can be correlated with the presence of pathogenic bacteria. For this reason the Italian regulation has defined limits for these microorganisms. The presence of these microbial indicators is dependent on various environmental variables. In this work all the variables considered are categorical and, consequently, the traditional approach of predictive microbiology was not applicable. The data were summarized by means of a cross-tabulation and analyzed using the log-linear model technique. This statistical technique is widely used in social and economic studies but only partially developed in food microbiology. The suitability of the log-linear model to analyse microbiological data in relation to environmental variables was evaluated. In particular, the microbiological quality of baby clams harvested in five different areas of the Adriatic Sea coast in Emilia Romagna (Italy) was considered. The influence of the season and geographical origin on microbiological standards was assessed. A logit model was developed to predict the frequencies, depending on geographical origin and season, of samples with concentrations of the indicator organisms below or above the legal standards provided by Italian regulation.

Animals↗

Effects of hexanal, trans-2-hexenal, and storage temperature on shelf life of fresh sliced apples.

In this paper, the effects of hexanal and trans-2-hexenal, which are both natural molecules characterizing apple aroma, on the microbial population and on color retention of fresh sliced apples were evaluated. In particular, a central composite design (CCD) was developed to assess the individual and interactive effects of the chosen volatile molecules and storage temperatures on (i) the growth of the naturally occurring microflora, (ii) the evolution over time of an inoculated spoilage yeast (Pichia subpelliculosa), and (iii) the enzymatic browning reaction in minimally processed apples. The inclusion of hexanal and trans-2-hexenal in the storage atmosphere of apple slices determined a significant extension of shelf life also when P. subpelliculosa was inoculated at levels of 10(3) colony-forming units/g and abusive storage temperatures were used. In fact, the presence of these molecules in the packaging atmospheres considerably prolonged the lag phases of the inoculated yeast and reduced the growth potential of naturally occurring bacteria. Moreover, the addition to the modified atmosphere of low levels of the hexanal increased the color stability of the products up to 16 days.

Aldehydes↗

Purification and characterization of an extracellular proline iminopeptidase from Arthrobacter nicotianae 9458.

An extracellular proline iminopeptidase, with a molecular mass of about 53 kDa, was purified from Arthrobacter nicotianae 9458 and characterized. The enzyme had temperature and pH optima of 37 degrees C and 8.0, respectively, was completely inactivated by heating for 1 min at 80 degrees C and showed highest activity on Pro-pNA. The proline iminopeptidase was characterized by activity at low temperature, NaCl concentrations up to 7.5% and by high sensitivity to pH values 6.0, serine enzyme inhibitor PMSF and divalent cations, Fe2+, Sn2+, Cu2+, Zn2+, Hg2+, Co2+ and Ni2+. The extracellular proline iminopeptidase from A. nicotianae 9458 was able to hydrolyze proline-containing peptides at the pH, temperature and NaCl concentration typical of the surface of smear-ripened cheese and may contribute to proteolysis of these cheeses during ripening.

Aminopeptidases↗

Effect of hexanal on the shelf life of fresh apple slices.

In this work the effects of hexanal, as a component of packaging atmosphere, on the shelf life of and evolution of naturally occurring microbial populations in fresh apple slices during storage at 4 and 15 degrees C were evaluated. Although hexanal had no bactericidal effects, in all conditions considered, this volatile molecule significantly extended the shelf life. In fact, the presence of hexanal in the storage atmosphere (at 4 degrees C) totally inhibited mesophilic bacteria and considerably prolonged the lag phase of psychrotrophic bacteria. Also, at 15 degrees C, hexanal strongly inhibited molds, yeasts, and mesophilic and psychrotrophic bacteria. Moreover, hexanal led to a yeast selection favoring species having a reduced spoilage potential due to their prevalent respiratory activity. When added to a modified atmosphere (70% N(2) and 30% CO(2)), this molecule was also very effective in preventing browning reactions for at least 16 days at 15 degrees C. No changes in hue angle values were observed in samples packaged in modified atmosphere with hexanal, even after 16 days of storage at 4 degrees C.

Aldehydes↗

Combined effects of ethanol, high homogenization pressure, and temperature on cell fatty acid composition in Saccharomyces cerevisiae.

The specific aims of this research were to evaluate the combined effects of ethanol and high-pressure homogenization at different temperatures on cell viability in Saccharomyces cerevisiae and to study the induced modification of fatty acid composition. The decrease in viability was weak at 10 degrees C while a homogenization pressure over 1000 bar (1 bar = 100 kPa) induced a significant reduction in viability when the cells were incubated at 20 and 30 degrees C. The cell tolerance to pressure decreased with an increase in ethanol concentration and temperature. Ethanol, particularly intracellular ethanol accumulated by S. cerevisiae, played an important role in the response to homogenization pressure and in modification of the cell fatty acid composition. In fact, an unusually elevated accumulation of ethyl esters in lipid extracts of yeast cells subjected to high homogenization pressure, especially in the presence of exogenous ethanol and at 30 degrees C, was observed. Moreover, only unsaturated and traces of short chain fatty acids were esterified with ethanol.

Esterification↗

Effect of high pressure homogenization on microbial and chemico-physical characteristics of goat cheeses.

The objective of this work was to compare goat cheeses obtained from milk previously subjected to high pressure homogenization (1000 bar) with those produced from untreated milk and milk subjected to sanitization (61 degrees C; 20 min) or to pasteurization (72 degrees C; 15 s). The pressure homogenization treatment had both direct and indirect effects on cheese characteristics and their evolution during ripening. The direct effects were principally linked to the change in water-binding capacity of proteins as shown also by the lower whey separation. The indirect effects involved the microbial growth or activity and, particularly, modifications of the population of the lactic acid bacteria that occurred naturally and their evolution as well as a more precocious yeast and mold growth with a consequent rapid rise in pH. Although the treatment proved to enhance both proteolytic and lipolytic activities according to Fourier transform infrared analysis, which was used to obtain a rapid description of the biochemical modification, the cheeses homogenized under high pressure showed relevant qualitative differences only in the zone corresponding to amide I and amide II signals of proteins. The activation of these enzymatic activities observed in the homogenized cheeses could be either an indirect effect of the shift of the microbial population or a consequence of a different exposure of the macromolecules to the enzymatic activity. Scanning electron microscopy analyses of goat cheeses revealed that cheeses homogenized under high pressure had a more homogeneous microstructure than did the others.

Animals↗

Entomologic and avian investigations of an epidemic of West Nile fever in Romania in 1996, with serologic and molecular characterization of a virus isolate from mosquitoes.

Between July and October 1996, a West Nile (WN) fever epidemic occurred in the southern plain and Danube Valley of Romania and in the capital city of Bucharest, resulting in hundreds of neurologic cases and 17 fatalities. In early October 1996, entomologic and avian investigations of the epidemic were conducted in the city of Bucharest and nearby rural areas. Thirty (41%) of 73 domestic fowl sampled had neutralizing antibody to WN virus, including 5 of 13 ducks (38%), 1 of 1 goose, 19 of 52 chickens (37%), 1 of 1 peahen, and 4 of 6 turkeys (67%). Seroprevalence in domestic fowl (27%, or 7 of 26) from the urban Bucharest site was not significantly different (P = 0.08, by Fisher's exact test) than rates at three rural sites (50%, or 23 of 46). Serum collected from one of 12 Passeriformes, an Erithacus rubecula, was positive for neutralizing antibody to WN virus. A total of 5,577 mosquitoes representing seven taxa were collected. Culex pipiens pipiens accounted for 96% of the mosquitoes collected. A single virus isolate, RO97-50, was obtained from a pool of 30 Cx. p. pipiens females aspirated from the walls and ceiling of a blockhouse located near the center of Bucharest, resulting in a minimum infection rate of 0.19 per 1,000. Antisera prepared against RO97-50 failed to distinguish among RO97-50, WN virus strain Eg101, and Kunjin (KUN) virus strain MRM16. A 2,323-basepair DNA fragment of the envelope (E) glycoprotein gene from RO97-50 and a Romanian WN virus strain obtained from a human cerebrospinal fluid sample, RO96-1030, were sequenced. Phylogenetic analyses of 23 WN virus strains and one KUN virus strain using the amino acid and nucleotide sequences for a small portion of the E gene suggest the existence of two large lineages of viruses. Bootstrap analysis of the nucleotide alignment indicated strong support (95%) for a lineage composed of WN virus strains from northern Africa, including isolates from Egypt and Algeria, and west, central, and east Africa, all of the European isolates, those from France and Romania, an Israeli isolate, and an isolate of KUN virus from Australia. The nucleotide sequence of RO97-50 was identical to the sequence of a WN virus isolate obtained from Cx. neavei mosquitoes from Senegal and Cx. univittatus mosquitoes from Kenya. The phylogenetic analyses were compatible with the introduction of virus into Romania by birds migrating from sub-Saharan Africa, to northern Africa, and into southern Europe.

Amino Acid Sequence↗