Intramuscular Immunization of Streptococcus pyogenes SF370 protein extract and identification of multiple virulence factors through proteomic profiling in RHD induced Balb/c mice.

Group A streptococcus (GAS) and autoimmunity are associated with heart related mitral valve damage, in adults. In this study Balb/c mice were intramuscularly immunized with S. pyogenes SF370 for 4 weeks. Prior to euthanization, physiological parameters like body weight and electrical signalling of the heart were recorded. After euthanization, the heart tissue homogenate was prepared and proteomic alterations were studied using SDS-PAGE and 2D electrophoresis. The expression levels of inflammatory genes like TNFα, IFNγ and TGF-ß were quantified using real time PCR. Insilico analysis was performed to identify the functions of hypothetical proteins and virulence factors involved in the induction of rheumatic carditis. The results showed a reduction in body weight, ulceration, inflammation, cardiac lesions and prolonged PR interval in mice immunized with S. pyogenes SF370, as a result of RHD. The heart related proteins like α-actinin, fatty acid binding protein-heart, myosin light chain 3, hemoglobin subunit alpha, myoglobin regulatory light chain 2, (ventricular/cardiac muscle isoform), myosin-6, troponin-1 were found to be up-regulated when compared with the control. The functional annotation of S. pyogenes (SF370) was carried out by retrieving 1696 identified proteins and 653 hypothetical protein sequences in NCBI genome database. The conserved domain was identified for 505 proteins. The pfam database documented that the super families of 279 sequences and 40 signal peptides enabled the classification of proteins in different categories like biological (20%), cellular (22%) and molecular functions (36%). Putative transcription repair coupling factor and putative lysine aminopeptidase N terminal are the two virulence factors identified by VICMPRED in S. pyogenes SF370. The two identified virulence factors are involved in altering the mice heart proteome and thereby controlling the streptococcus pyogenes infection. Thus, the results of the present study reveals the role of immunogenic proteins in induction of rheumatic carditis and to elucidate the molecular mechanisms leading to autoimmune reactions in Balb/c mice.

Abiotic stress hormesis: An approach to maintain quality, extend storability, and enhance phytochemicals on fresh produce during postharvest.

Postharvest losses of whole and fresh-cut fruits and vegetables cause significant reductions in food availability and an increase in economic losses/damages. Additionally, regulatory agencies are increasingly restricting the postharvest use of synthetic chemicals. This has strengthened the need to develop environmentally friendly approaches to postharvest management, such as utilization of natural compounds, antagonist microorganisms, and treatments with abiotic stresses, among others. The current review focuses on the potential of low doses of abiotic stresses to extend the shelf life, increase the amount of health beneficial phytochemicals, and reduce postharvest losses of fresh produce. The positive effects of the responses to low doses of abiotic stresses are based on a biological phenomenon termed hormesis. Research to develop new technologies to improve postharvest handling of fresh fruit and vegetables as well as minimally processed products is critical. The phenomenon of abiotic stress hormesis in fresh fruit and vegetables shows the potential not only to enhance defense compounds that could reduce diseases during postharvest storage and extend shelf life but also to elevate the content of health-promoting substances. The beneficial effects of UV-C hormesis have been extensively investigated in numerous types of fresh produce. However, our knowledge on hormesis exhibited by other abiotic stresses is still limited. Hence, the objective of this review is to discuss the relevance of hormesis for postharvest research by examining whether all abiotic stresses exhibit the phenomenon, its biological significance, the potential application in various commodities, and how it may direct the future of postharvest research.

In vitro protection of biological macromolecules against oxidative stress and in vivo toxicity evaluation of Acacia nilotica (L.) and ethyl gallate in rats.

BACKGROUND: Recently, enormous research has been focused on natural bioactive compounds possessing potential antioxidant and anticancer properties using cell lines and animal models. Acacia nilotica (L.) is widely distributed in Asia, Africa, Australia and Kenya. The plant is traditionally used to treat mouth, ear and bone cancer. However, reports on Acacia nilotica (L.) Wild. Ex. Delile subsp. indica (Benth.) Brenan regarding its toxicity profile is limited. Hence in this study, we investigated the antioxidant capacity and acute toxicity of ethyl gallate, a phenolic antioxidant present in the A. nilotica (L.) leaf extract. METHODS: The antioxidant activity of ethyl gallate against Fenton's system (Fe3+/H2O2/ascorbic acid) generated oxidative damage to pBR322 DNA and BSA was investigated. We also studied the interaction of ethyl gallate to CT-DNA by wave scan and FTIR analysis. The amount of ethyl gallate present in the A. nilotica (L.) leaf extract was calculated using HPLC and represented in gram equivalence of ethyl gallate. The acute toxicity profile of ethyl gallate in the A. nilotica (L.) leaf extract was analyzed in albino Wistar rats. Measurement of liver and kidney function markers, total proteins and glucose were determined in the serum. Statistical analysis was done using statistical package for social sciences (SPSS) tool version 16.0. RESULTS: Ethyl gallate was found to be effective at 100 µg/mL concentration by inhibiting the free radical mediated damage to BSA and pBR322 DNA. We also found that the interaction of ethyl gallate and A. nilotica (L.) leaf extract to CT-DNA occurs through intercalation. One gram of A. nilotica (L.) leaf extract was found to be equivalent to 20 mg of ethyl gallate through HPLC analysis. Based on the acute toxicity results, A. nilotica (L.) leaf extract and ethyl gallate as well was found to be non-toxic and safe. CONCLUSIONS: Results revealed no mortality or abnormal biochemical changes in vivo and the protective effect of A. nilotica (L.) leaf extract and ethyl gallate on DNA and protein against oxidative stress in vitro. Hence, A. nilotica (L.) leaf extract or ethyl gallate could be used as potential antioxidants with safe therapeutic application in cancer chemotherapy.

The role of hydrolases in the loss of firmness and of the changes in sugar content during the post-harvest maturation of Carica papaya L. var solo 8.

Fruit ripening is associated with many hydrolase activities involved in the softening of the fruit during the maturation. This study investigates the relationship between the loss of firmness along with the changes of sugar content and the enzymatic activities in Carica papaya L.var solo 8 during post-harvest storage. Three maturation stages (green immature: the fruit is entirely green, green mature: the fruit shows 1/32 yellow skin and fully mature: the fruit shows 1/8 yellow skin) have been selected and stored at 15, 22 and 28 °C. The reduction of fruit firmness, total sugar contents, refractive index (% Brix) and enzymatic activities were measured. Low enzymatic activities (0.035 µmol/min/mg) were recorded in fruit harvested at the green immature stage with no significant (p ≥ 0.05) effect on the softening while fruit harvested at the green mature and fully mature stages showed enzymatic activities 7 times as high as those of the green immature stage. These high enzymatic activities were responsible for the loss of firmness of the fruit. Accordingly, papayas at the green mature and fully mature stages displayed higher maxima of sugar content (4.8 g/100 g at 28 °C at day 12, and 10.2 g/100 g at 22 °C at day 8, respectively) at higher temperatures. Meanwhile in green immature papayas, the maximum was only 4.3 g/100 g at 22 °C and day 12 of storage. The results show that the loss of firmness of the papaya was highly related to the hydrolytic enzyme activities and the sweet taste to the presence of simple sugars such as galactose liberated from the polysaccharide complexes.

Fragmentation of chitosan by acids.

Fragmentation of chitosan in aqueous solution by hydrochloric acid was investigated. The kinetics of fragmentation, the number of chain scissions, and polydispersity of the fragments were followed by viscometry and size exclusion chromatography. The chemical structure and the degree of N-acetylation (DA) of the original chitosan and its fragments were examined by (1)H NMR spectroscopy and elemental analysis. The kinetic data indicates that the reaction was of first order. The results of polydispersity and the DA suggest that the selected experimental conditions (temperature and concentration of acid) were appropriate to obtain the fragments having the polydispersity and the DA similar to or slightly different from those of the original one. A procedure to estimate molecular weight of fragments as well as the number of chain scissions of the fragments under the experimental conditions was also proposed.

Phytochemical Enhancement in Broccoli Florets after Harvest by Controlled Doses of Ozone.

The objective of this work was to examine the effect of controlled doses of O3 (0, 5 µL L-1 of O3 for 60 min, and 5 µL L-1 of O3 for 720 min) on the quality and phytochemical content of broccoli florets during postharvest storage. The optimal dose was found at 5 µL L-1 of O3 for 60 min, from the color retention of broccoli florets exposed to the gas treatment. Overall, the antioxidant capacity of the florets was significantly affected by both doses of O3 compared to the non-exposed florets. The profile of glucosinolates was determined for up to 14 days in broccoli florets stored at 4 °C by LC-MS. The amount of total glucobrassicins and total hydroxy-cinnamates in florets significantly (p ≤ 0.05) improved by the application of 5 µL L-1 of O3 for 60 min compared to non-treated florets. The up-regulation of genes of the tryptophan-derived glucosinolate pathway was observed immediately after both treatments. The gene expression of CYP79A2 and CYP79B3 in broccoli was significantly higher in broccoli florets exposed to 5 µL L-1 of O3 for 720 min compared to non-exposed florets. Although enhancement of secondary metabolites can be achieved by the fumigation of broccoli florets with low doses of ozone, quality parameters, particularly weight loss, can be compromised.

Physicochemical basis for the inhibitory effects of organic and inorganic salts on the growth of Pectobacterium carotovorum subsp. carotovorum and Pectobacterium atrosepticum.

Twenty-one salts were tested for their effects on the growth of Pectobacterium carotovorum subsp. carotovorum and Pectobacterium atrosepticum. In liquid medium, 11 salts (0.2 M) exhibited strong inhibition of bacterial growth. The inhibitory action of salts relates to the water-ionizing capacity and the lipophilicity of their constituent ions.

The -866G/A polymorphism in the promoter of the UCP2 gene is associated with risk for type 2 diabetes and with decreased insulin levels.

BACKGROUND AND AIM: Oxidative stress and impaired insulin secretion is an underlying major risk factor for the development of type 2 diabetes (T2D). Uncoupling protein-2 (UCP2) is involved in the regulation of reactive oxygen species production, insulin secretion, and lipid metabolism. Based on this we aimed to find an association of UCP2 (G-866A) polymorphism with the risk of T2D in South Indian population. METHODS: A total of 318 T2D patients and 312 controls were enrolled in this study. All the study subjects were genotyped for UCP2 (G-866A) polymorphism using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Fasting blood glucose, HbA1c, serum lipid profile, systolic and diastolic blood pressure were measured by standard biochemical methods. Fasting serum insulin level was measured by ELISA. RESULTS: In UCP2 (G-866A) polymorphism, the distribution of GA (46%) and AA (14%) genotypes were significantly higher in T2D patients than the healthy controls. The frequency of GA and AA genotypes have high risk towards the development of T2D with an Odds Ratio (OR) of 1.55 (P = 0.01) and 2.04 (P = 0.01) respectively. Moreover, SNP-866 G>A allele was found to be significantly associated with T2D (OR = 1.48, P = 0.001, 95% CI = 1.16-1.88). Further, the UCP2 AA genotype showed significantly decreased level of insulin by the reduction in pancreatic ß-cell function in T2D patients. CONCLUSION: UCP2 (G-866A) polymorphism may play a crucial role in the pathogenesis of insulin secretion thus leads to the development of T2D.

Fragmentation of chitosan by ultrasonic irradiation.

Kinetics of chitosan fragmentation by ultrasonic irradiation at frequency of 20 kHz, and the effects of experimental variables (power of ultrasound, chitosan concentration and solution temperature) on fragmentation were investigated. The kinetics studies were followed by measuring solution viscosity of the original and its fragments, and determining average number of chain scission of the fragments. The effects of ultrasonic power, chitosan concentration and solution temperature on fragmentation process were followed by viscometry and size exclusion chromatography. The chemical structure of the original chitosan and its fragments were examined by (1)H NMR spectroscopy and elemental analysis. The experimental results showed that the rate of fragmentation increased with an increase in power of ultrasound. Chain scission increased with an increase in power of ultrasound; and solution temperature, but a decrease in chitosan concentration. The chemical structure and polydispersity of the original and the fragments were nearly identical. A model based on experimental data to describe the relationship between chain scission and experimental variables (power of ultrasound; irradiation time; reduced concentration, c[eta]; and solution temperature) was proposed. It was concluded that ultrasonic irradiation is a suitable method to perform partial depolymerization and to obtain moderate macromolecules from large ones.

PRMT1 promotes hyperglycemia in a FoxO1-dependent manner, affecting glucose metabolism, during hypobaric hypoxia exposure, in rat model.

PURPOSE: High-altitude (HA) environment causes changes in cellular metabolism among unacclimatized humans. Previous studies have revealed that insulin-dependent activation of protein kinase B (Akt) regulates metabolic processes via discrete transcriptional effectors. Moreover, protein arginine methyltransferase (PRMT)1-dependent arginine modification of forkhead box other (FoxO)1 protein interferes with Akt-dependent phosphorylation. The present study was undertaken to test the involvement of PRMT1 on FoxO1 activation during hypobaric hypoxia (HH) exposure in rat model. METHODS: Samples were obtained from normoxia control (NC) and HH-exposed (H) rats, subdivided according to the duration of HH exposure. To explore the specific role played by PRMT1 during HH exposure, samples from 1d pair-fed (PF) NC, 1d acute hypoxia-exposed (AH) placebo-treated, and 1d AH TC-E-5003-treated rats were investigated. Quantitative reverse transcriptase polymerase chain reaction (RT-qPCR) was performed to determine expressions of glycolytic, gluconeogenic enzymes, and insulin response regulating genes. Immuno-blot and enzyme linked immunosorbent assay (ELISA) were used for insulin response regulating proteins. Nuclear translocation of FoxO1 was analyzed using deoxyribonucleic acid (DNA)-binding ELISA kit. RESULTS: We observed HH-induced increase in glycolytic enzyme expressions in hepatic tissue unlike hypothalamic tissue. PRMT1 expression increased during HH exposure, causing insulin resistance and resulting increase in FoxO1 nuclear translocation, leading to hyperglycemia. Conversely, PRMT1 inhibitor treatment promoted inhibition of FoxO1 activity and increase in glucose uptake during HH exposure leading to reduction in blood-glucose and hepatic glycogen levels. CONCLUSIONS: PRMT1 might have a potential importance as a therapeutic target for the treatment of HH-induced maladies.

Association of paraoxonase-1 gene polymorphisms with insulin resistance in South Indian population.

BACKGROUND AND AIM: Insulin resistance plays a crucial role in the pathogenesis of type 2 diabetes and cardiovascular diseases. Recently, paraoxonase-1(PON1) is reported to have an ability to reduce insulin resistance by promoting glucose transporter-4 (GLUT-4) expression in vitro. Single nucleotide polymorphism (SNP) in PON1 is associated with variability in enzyme activity and concentration. Based on this we aimed to investigate the association of PON1 (Q192R and L55M) polymorphisms with the risk of developing insulin resistance in adult South Indian population. METHODS: Two hundred and eighty seven (287) Type 2 diabetes patients and 293 healthy controls were enrolled in this study. All the study subjects were genotyped for PON1 (Q192R and L55M) missense polymorphisms using polymerase chain reaction-restriction fragment length polymorphism (PCRRFLP) method. Fasting serum insulin level was measured by ELISA. RESULTS: The distribution of QR/RR and LM/MM genotypes were significantly higher in type 2 diabetes patients compared with healthy controls. Moreover, the R and M alleles were significantly associated with type 2 diabetes with an Odds Ratio of 1.68 (P < 0.005) and 2.24 (P < 0.005) respectively. SNP 192 Q > R genotypes were found to be significantly associated with higher BMI, cholesterol, triglycerides, LDL, fasting serum insulin and HOMA-IR. Further, the mutant allele or genotypes of PON1 L55M were associated with higher BMI, triglycerides, VLDL, fasting serum insulin and HOMA-IR among adult type 2 diabetes patients. CONCLUSION: PON1 (Q192R and L55M) polymorphisms may play a crucial role in pathogenesis and susceptibility of insulin resistance thus leads to the development of type 2 diabetes in South Indian population.

Electromigration behavior of a mixture of chitosan oligomers at different concentrations.

In this study, the effect of the concentration of a chitosan oligomer mixture on its electrophoretic behavior was studied as a function of pH and ionic strength added. It was shown that the concentration has a significant effect on the average electrophoretic mobility of the chitosan oligomer mixture and on isoelectric point. At a concentration of 3%, the ionic strength added did not show any effect on the electromigration behavior of the chitosan oligomer mixture. By decreasing the concentration of the chitosan oligomer mixture, ionic strength showed a significant effect on the average electrophoretic mobility but not on the isoelectric point. The highest shift of the isoelectric point was recorded in water at 0.003% concentration of the oligomer mixture. Under these conditions, the isoelectric point was at pH 5 whereas it was at pH 8 at 3% concentration of chitosan oligomer mixture. Electrophoretic measurements were also taken in water/ethanol aqueous medium. By adding ethanol to the medium, the average electrophoretic mobility decreased. This would have been caused by the increase in viscosity of the medium. Increasing ethanol ratio in the running medium, the isoelectric point moved from pH 5 in water up to pH 6-8 dependently on chitosan oligomer mixture concentration and ethanol content of the medium.

Electromigration of chitosan D-glucosamine and oligomers in dilute aqueous solutions.

The electromigration behavior of chitosan D-glucosamine and oligomers with a degree of polymerization from 1 to 6 in dilute aqueous systems containing either NaCl or KCl salt at 0.01, 0.05, and 0.1 M at pH values from 2 to 9 was evaluated. The results showed that the electromigration of the chitosan D-glucosamine and oligomers did not change by changing the type of salt in the running medium and that the pH had a significant effect on the direction of migration under an external electric field. In addition, the increase in the ionic strength of the medium caused a significant decrease on the absolute value of the electrophoretic mobility, and the highest values of the electromobility were observed in water. However, the ionic strength had no significant effect on the electrophoretic mobilities at pH 2 in comparison with the other pH values. The dimer showed the highest electrophoretic mobility in the alkaline zone of the pH. At pH values lower than the pKa of the D-glucosamine, the chitosan D-glucosamine, and oligomers migrated toward the anode, where the amine groups are protonated and carry positive charge. At higher pH values, chitosan D-glucosamine and oligomers migrated toward the anode, even though they did not carry any electric charge. The contribution of the difference in the dielectric constants between the solvent and the solute to this phenomenon was highlighted. It was shown that the glucose moiety contributes to the direction of migration of the chitosan D-glucosamine and oligomers under alkaline conditions and that the difference in the dielectric constant of glucose and the solvent accounts for the direction and the extent of electromobility.

Solubilization of chitosan by bipolar membrane electroacidification.

Chitosan, a partially deacetylated derivative of chitin, was solubilized by bipolar membrane electroacidification (BMEA). Bipolar/monopolar (anionic or cationic) configuration and chitosan addition mode (single step or stepwise) were examined. Chitosan solubility and electroacidification parameters were monitored during the process to determine the optimal conditions. Bipolar/anionic configuration and stepwise feeding mode led to chitosan solubilization yield of 91% in 60 min at 20 mA/cm(2). In this configuration, chitosan solution had a pH of 2.5, a conductivity of 8.5 mS/cm, and an ash content of 0.2%. Relative energy consumption was 0.05 kWh/L of 1% chitosan solution prepared. Although some chitosan particles were aggregated in the electrodialysis stack, limiting chitosan solubilization, BMEA allowed complete solubilization of chitosan circulating in the system.

Determination of N-nitrosamines in processed meats by liquid extraction combined with gas chromatography-methanol chemical ionisation/mass spectrometry.

A simple, accessible and reproducible method was developed and validated as an alternative for the determination of nine volatile N-nitrosamines (NAs) in meat products, using a low volume of organic solvent and without requiring specific apparatus, offering the possibility of practical implementation in routine laboratories. The NAs were extracted with dichloromethane followed by a clean-up with phosphate buffer solution (pH 7.0). The extracts were analysed by gas chromatography-chemical ionisation/mass spectrometry (GC-CI/MS) in positive-ion mode using methanol as reagent. Limits of detection and quantification, recovery and reproducibility were determined for all NAs (N-nitrosodimethylamine, N-nitrosomethylethylamine, N-nitrosodiethylamine, N-nitrosopyrrolidine, N-nitrosodipropylamine, N-nitrosomorpholine, N-nitrosopiperidine, N-nitrosodibutylamine and N-nitrosodiphenylamine). Satisfactory sensitivity and selectivity were obtained even without concentrating the extract by solvent evaporation, avoiding the loss of the nine NAs studied. Limits of detection ranged from 0.15 to 0.37 µg kg(-1), whereas limits of quantification ranged from 0.50 to 1.24 µg kg(-1). Recoveries calculated in cooked ham that had been spiked at 10 and 100 µg kg(-1) were found to be between 70% and 114% with an average relative standard deviation of 13.2%. The method was successfully used to analyse five samples of processed meat products on the day of purchase and 7 days later (after storage at 4°C). The most abundant NAs found in the analysed products were N-nitrosodipropylamine and N-nitrosopiperidine, which ranged from 1.75 to 34.75 µg kg(-1) and from 1.50 to 4.26 µg kg(-1), respectively. In general, an increase in the level of NAs was observed after the storage period. The proposed method may therefore be a useful tool for food safety control once it allows assessing the profile and the dietary intake of NAs in food over time.

Changes in ghrelin, CCK, GLP-1, and peroxisome proliferator-activated receptors in a hypoxia-induced anorexia rat model.

INTRODUCTION: A high-altitude environment causes appetite loss in unacclimatised humans, leading to weight reduction. Ghrelin, cholecystokinin (CCK), and glucagon like peptide-1 (GLP-1), are gut hormones involved in the regulation of food intake and energy metabolism. The liver is an important site of metabolic regulation, and together with the gut it plays a role in food intake regulation. This study intends to study the time-dependent changes occurring in plasma gut hormones, PPARα, PPARδ, and PGC1α, in the stomach and liver during hypoxia. MATERIAL AND METHODS: Male Sprague Dawley rats were exposed to hypobaric hypoxia in a decompression chamber at 7620 m for different durations up to seven days. RESULTS: Hypoxia increased circulating ghrelin from the third day onwards while CCK and GLP-1 decreased immediately. An increase in ghrelin, ghrelin receptor protein levels, and GOAT mRNA levels in the stomach was observed. Stomach cholecystokinin receptor (CCKAR), PPARα, and PPARδ decreased. Liver CCKAR decreased during the first day of hypoxia and returned to normal levels from the third day onwards. PPARα and PGC1α expression increased while PPARδ protein levels reduced in the liver on third day. CONCLUSION: Hypoxia alters the expression of ghrelin and ghrelin receptor in the stomach, CCKAR in the liver, and PPAR and its cofactors, which might be possible role players in the contribution of gut and liver to anorexia at high altitude.

Role of BMP7 in appetite regulation, adipogenesis, and energy expenditure.

Bone morphogenetic protein 7 (BMP7), also known as osteogenic protein-1 (OP-1) is a member of Transforming growth factor-ß (TGF-ß) family of proteins. Bone morphogenetic proteins were discovered in 1965 by Marshal Urist, of which BMP7 is of particular interest in this review being a leptin-independent anorexinogen and having role in energy expenditure in the brown adipose tissue, which makes it a potential target for preventing/treating obesity. As it has been established that Obesity displays a state of leptin-resistance, thus a protein-like BMP7 which acts through a leptin-independent pathway could give new therapeutic directions. This review will also discuss the synthesis and action of BMP7, along with its receptors and signal transduction. A brief note about BMP7-mediated brown fat development and energy balance is also discussed.

Fasting and postprandial lipid response to the consumption of modified milk fats by guinea pigs.

The objective of the present study was to investigate the effect of three modified milk fats with different melting profiles on fasting and postprandial lipid responses and on fecal fat content in guinea pigs. We hypothesized that the consumption of modified milk fat with a high m.p. results in reduced fasting and postprandial lipid responses compared with that of modified milk fat fractions with lower m.p. To test this hypothesis, male Hartley guinea pigs were fed isoenergetic diets containing 110 g of fat/kg, either from one of the three modified milk fats with high (HMF), medium (MMF), or low melting profiles (LMF), or from one of the two reference fats as whole milk fat (MF) or a fat blend similar to that of nonhydrogenated soft margarine (MA) for 28 d. Food intake (P < 0.05) and body weight gain (P < 0.05) were reduced in the animals fed the HMF diet compared with the other groups. In the fasting state, plasma LDL cholesterol was highest in animals fed the LMF diet, intermediary in those fed the MMF and MF diets, and lowest in those fed the HMF and MA diets (P< 0.05). Postprandially, the areas under the 0- to 3-h curves for the changes in plasma TG were lower in the HMF group than in the MA- and LMF-fed guinea pigs (P< 0.05). The fecal fat content was higher (P< 0.05) in the HMF group compared to the other milk fat groups. The present results suggest that modified milk fats can impact food intake, body weight gain, fasting cholesterolemia, and postprandial triglyceridemia, and these changes may be attributed to an altered fat absorption.

Dibutyrate derivatization of monoacylglycerols for the resolution of regioisomers of oleic, petroselinic, and cis-vaccenic acids.

Dibutyrate derivatives of monoacylglycerols of oleic, petroselinic, and cis-vaccenic acids were prepared by diesterification of monoacylglycerols with n-butyryl chloride. The resulting triacylglycerols were analyzed by gas chromatography (GC) with a 65% phenyl methyl silicone capillary column and separated on the basis of both fatty acid composition and regiospecific position. The petroselinic acid derivatives eluted first, followed sequentially by the oleic and cis-vaccenic acid derivatives, with the sn-2 positional isomer eluting before the sn-1 (3) isomer in each case. Separation of the peaks was almost baseline between petroselinic and oleic acids as well as between oleic and cis-vaccenic acids. To assess the accuracy of the method, mixtures of triolein, tripetroselinin, and tri-cis-vaccenin in various known proportions were partially deacylated with the use of ethyl magnesium bromide and derivatized and analyzed as above. The results showed that this method compares favorably to the existing methods for analysis of oleic, petroselinic, and cis-vaccenic fatty acids by GC with respect to peak separation and accuracy, and it also provides information on the regiospecific distribution of the fatty acids. The method was applied to basil (Ocimum basilicum) and coriander (Coriandrum sativum) seed oils. cis-Vaccenic, oleic, and linoleic acids were mainly distributed at the sn-2 position in basil seed oil, and higher proportions of linolenic, palmitic, and stearic acids were distributed at the sn-1(3) position than at the sn-2 position. In coriander seed oil, petroselinic acid was mainly distributed at the sn-1 (3) position, and both oleic and linoleic acids were mostly located at the sn-2 position, whereas palmitic, stearic, and cis-vaccenic acids were located only at the sn-1 (3) position.

Effect of water activity on the production of volatile organic compounds by Muscodor albus and their effect on three pathogens in stored potato.

Muscodor albus (Xylariaceae, Ascomycetes) isolate CZ-620 produces antimicrobial volatile organic compounds (VOC), which appear to have potential for the control of various postharvest diseases. The effect of water activity (Aw) on the production of VOC by M. albus culture, and their inhibitory effects on the growth of three pathogens of potato tuber (Fusarium sambucinum, Helminthosporium solani, and Pectobacterium atrosepticum) and the development of diseases caused by the three pathogens (dry rot, silver scurf, and bacterial soft rot, respectively) were investigated. Rye grain culture of the fungus produced six alcohols, three aldehydes, five acids or esters, and two terpenoids. The most abundant VOC were: isobutyric acid; bulnesene, a sesquiterpene; an unidentified terpene; 2 and 3-methyl-1-butanol; and ethanol. However, the level of each of those VOC varied with Aw of the culture. Emission activity occurred mainly at Aw above 0.75 and high emission of most VOC occurred only at Aw above 0.90. The aldehydes (2-methyl-propanal and 3-methyl-butanal) were the only VOC produced in quantities below an Aw of 0.90. An Aw value of 0.96 favored maximum emission of acids, esters, and terpenoids. There was a higher production of alcohols and a decrease in aldehydes with increase in Aw. Isobutyric acid, which has been the main M. albus VOC monitored in previous studies as an indicator of antifungal activity, had a rather narrow optimum, peaking at Aw of 0.96 and declining sharply above 0.98. Results showed that substrate Aw affects the production dynamics of each group of VOC by the fungus, and suggest that VOC production can be prolonged by maintaining M. albus culture at a constant optimum Aw. The VOC was inhibitory to F. sambucinum, H. solani, and P. atrosepticum; and biofumigation with M. albus significantly reduced dry rot and soft rot development, and completely controlled silver scurf in inoculated tubers incubated at both 8°C and 22°C. The results show that Aw of grain culture affects the production of VOC by M. albus; and that the VOC inhibit the growth of the tested pathogens and the diseases caused by them in potato tubers.