Quercetin repressed the stress response factor (sigB) and virulence genes (prfA, actA, inlA, and inlC), lower the adhesion, and biofilm development of L. monocytogenes.

The present study explored the effect of quercetin on the expression of virulence genes actA, inlA, inlC, and their regulatory components, sigB and prfA, in L. monocytogenes. Furthermore, the physicochemical changes on the surface, membrane permeability, and biofilm formation of quercetin-treated bacteria were evaluated. An inhibitory dose-dependent effect of quercetin (0.1-0.8 mM) was observed on the cell attachment on stainless steel at 2 and 6 h at 37 °C. Quercetin at 0.8 mM prevented the biofilm formation on stainless steel surfaces after 6 h of incubation at 37 °C, while the untreated bacteria formed biofilms with a cell density of 5.1 Log CFU/cm2. The microscopic analysis evidenced that quercetin at 0.2 mM decreased the biovolume and covered area of the attached micro-colonies. Also, sigB, prfA, inlA, inlC, and actA genes were downregulated by 7-29 times lower compared to untreated bacteria. In addition, quercetin decreased the superficial cell charge, increased the membrane permeability, and its surface hydrophobicity. These results demonstrated that quercetin prevented biofilm formation, repressed the genes of stress and virulence of L. monocytogenes and also altered the physicochemical cell properties.

Morphometric parameters of foodborne related-pathogens estimated by transmission electron microscopy and their relation to optical density and colony forming units.

The different morphological characteristics of five bacterial pathogen strains were analyzed through transmission electron microscopy for addressing the particular relationship between optical density and colony-forming units for each strain. Generated linear equations will allow a reliable calculation of bacterial concentrations through simple optical density measurements.

Surface disinfection procedure and in vitro regeneration of grapevine (Vitis vinifera L.) axillary buds.

Establishment of an efficient explants surface disinfection protocol is essential for in vitro cell and tissue culture as well as germplasm conservation, such as the case of Grapevine (Vitis spp.) culture. In this research, different procedures for disinfection and regeneration of field-grown grapevine cv. 'Flame seedless' axillary buds were evaluated. The buds were disinfected using either NaOCl or allyl, benzyl, phenyl and 2-phenylethyl isothiocyanates. Two different media for shooting and four media for rooting were tested. Shoot and root development per buds were registered. The best disinfection procedure with 90 % of tissue survival involved shaking for 60 min in a solution containing 20 % Clorox with 50 drops/L Triton(®) X-100. These tissues showed the potential to regenerate a complete plant. Plant regeneration was conducted using full strength Murashigue and Skoog (MS) medium supplemented with 8 µM benzyl aminopurine for shoot induction and multiplication, whereas rooting was obtained on half strength MS supplemented with 2 mg L(-1) of indole-3-butyric acid and 200 mg L(-1) of activated charcoal. In this work, it was designed the protocols for obtaining sterile field-grown grapevine buds and in vitro plant development. This methodology showed potential to produce vigorous and healthy plants in 5 weeks for clonal grapevine propagation. Regenerated plants were successfully established in soil.

In vitro antiviral activity against rotavirus and astrovirus infection exerted by substances obtained from Achyrocline bogotensis (Kunth) DC. (Compositae).

BACKGROUND: Achyrocline bogotensis has been traditionally used to treat infections of skin, respiratory, tract urinary and other infections, but not to treat viral gastrointestinal disease. In this study, this Colombian native medicinal plant was investigated by its in vitro anti-rotavirus and anti-astrovirus activity. METHODS: Several extracts and fractions phytochemically obtained from A. bogotensis were evaluated initially for their cell toxicity on MA104 and Caco2 cells and then for their anti-rotavirus (RRV) and anti-astrovirus (Yuc8) activity following three strategies: pre-treatment of cells (blocking effect), direct viral activity (virucidal effect) and post-treatment of infected cells (reduction of viral yield post-infection). In addition qualitative chemical studies were developed for the active compounds. RESULTS: Non-toxic concentrations of a fraction obtained exhibited antiviral activity against both viruses characterized by a virucidal effect and by the reduction of the infectious particles produced post-infection. Steroids, sterols, terpenes, phenols, flavonoids and sesquiterpenlactones were identified qualitatively in the active fraction. CONCLUSIONS: A. bogotensis contains substances with in vitro antiviral activity against rotavirus and astrovirus. This study confirms their anti-microbial properties and describes by the first time its antiviral activity in vitro.

Semiochemicals released by pecan alleviate physiological suppression in overwintering larvae of Acrobasis nuxvorella (Lepidoptera: Pyralidae).

Acrobasis nuxvorella Neunzig (pecan nut casebearer) is a monophagous herbivore of Carya illinoinensis (Wang.) K. Koch (pecan); both are indigenous to North America, where Carya has evolved for ≈60 million years. We hypothesized that this close association may have resulted in a parallel evolution allowing casebearer to use pecan volatiles to synchronize seasonality. Casebearer overwinters in diapause as a first-instar larva in a hibernaculum attached to a dormant pecan bud. Larval emergence from this structure after diapause or postdiapause quiescence coincides with the onset of pecan bud growth in the spring, and this interaction was the subject of this study. Dormant pecan twigs with hibernacula-infested buds were exposed to a water control or pecan volatiles from 'Western Schley' cultivar, and monitored to observe larval response by using a microcalorimeter. Initial testing showed that metabolic heat produced by overwintering larvae remained low and unchanged when exposed to water vapor and significantly increased within a few hours after exposure to volatiles from new pecan foliage. This shows that these larvae in hibernacula are in a physiologically suppressed state of diapause or postdiapause quiescence, from which they detect and respond to these pecan volatiles. Further studies to quantify larval responses showed that 90 and 80% of the larvae became active and emerged from their hibernacula ≈6 d after exposure to Western Schley and 'Wichita' volatiles, respectively. Mixtures of 13 sesquiterpenes from those pecan volatiles were identified to induce physiological activity within larvae after hours of exposure, followed some days later by larval emergence from hibernacula. Host volatiles, to our knowledge, have not previously been reported to induce early instar larvae in hibernacula to rouse from a state of physiological arrest to resume normal growth and development. This also has potential for use in pest management.

Improving antioxidant capacity of fresh-cut mangoes treated with UV-C.

The effect of UV-C irradiation time on total phenol, flavonoids, beta-carotene, ascorbic acid contents, and antioxidant capacity (ORAC, DPPH(*)) of fresh-cut "Tommy Atkins" mango stored for 15 d at 5 degrees C was investigated. Fresh-cut mango was irradiated for 0, 1, 3, 5, and 10 min prior to storage at 5 degrees C. UV-C irradiation for 10 min induced a hypersensitive defense response resulting in the phenols and flavonoids accumulation which was positively correlated with ORAC and DPPH(*) values. However, beta-carotene and ascorbic acid content of fresh-cut mangos decreased with irradiation time during storage. Antioxidant capacity (ORAC, DPPH(*)) was increased in fresh-cut mangoes treated with UV-C irradiation. In conclusion, UV-C irradiation appears to be a good technique to improve the total antioxidant capacity of fresh-cut mango.

PCR identification of Salmonella: potential contamination sources from production and postharvest handling of cantaloupes.

Salmonella is one of the most frequently reported etiological agents in outbreaks of foodborne diseases associated with the consumption of cantaloupes. Sensitive and reliable methods for detecting and identifying foodborne microorganisms are needed. The PCR can be used to amplify specific DNA fragments and thus to detect and identify pathogenic bacteria. In this study, a PCR method was used to evaluate the incidence of Salmonella at cantaloupe production, harvest, and packaging steps, and the results were compared with those of the standard method for detection of Salmonella in foods (Mexican NOM-114-SSA1-1994). Salmonella was detected by both standard and PCR methods in 23.5% of the irrigation water samples but only by the PCR method in 9.1% of the groundwater samples, 4.8% of the chlorinated water samples, 16.7% of samples from the hands of packing workers, 20.6% of samples from the packed cantaloupes, and 25.7% of samples from the in-field cantaloupes. With the standard method, Salmonella was found in 8.3% of the crop soil samples. Statistical analysis indicated a significant difference in sensitivity (P < 0.05) between the two methods; the PCR method was 4.3 times more sensitive than the standard method. Salmonella was found at seven of the eight pointsevaluated during the production and postharvest handling of cantaloupe melons.

Methyl jasmonate treatments reduce chilling injury and activate the defense response of guava fruits.

Tropical fruits cannot be stored at low temperatures due to the chilling injury phenomena. With the goal of reducing the chilling injury, we tested 10(-4) and 10(-5) M of methyl jasmonate (MJ) treatment before the storage of red and white cultivars of guava fruits at 5 degrees C for up to 15 days plus two days at 20 degrees C. Every five days, we evaluated chilling injury index, ion leakage percentage, vitamin C, sugars, total phenols, and the activity of the enzymes lipoxygenase (LOX) and phenylalanine-ammonia lyase (PAL). We found that methyl jasmonate treatments reduce the chilling injury index and the ion leakage percentage. Furthermore, MJ did not affect vitamin C, chlorophyll, and total phenols. MJ increased sugar content, PAL, and LOX activities. We concluded that MJ reduces chilling injury and activates the fruit defense response as indicated by the behavior of total phenols and the increase in sugar content, PAL, and LOX activities.

Effect of infiltrated polyamines on polygalacturonase activity and chilling injury responses in zucchini squash (Cucurbita pepo L.).

The effect of exogenous polyamines on electrolyte leakage, chilling index, polygalacturonase activity (PG), ethylene production, and firmness in zucchini squash fruits stored for 12 days at 2 degrees C or 10 degrees C, 85-90% RH was evaluated. Fruits were infiltrated with putrescine (PUT) spermidine (SPD) and spermine (SPM) at 0.1, 0.25, 0.5, 2.0, and 4.0 mM. All polyamines exerted a protective effect on cell and organelle membranes. The most effective was SPD, which reduced electrolyte leakage between 62% and 82%, compared to control fruits stored at 2 degrees C. At 10 degrees C they did not exhibit chilling injury (CI) symptoms, while at 2 degrees C SPM (0.5 mM) and SPD (0.5 mM) diminished them 92% and 100%, respectively; which extended storage life for 8-10 days at 2 degrees C. High concentrations of polyamines (>2.0 mM) caused the appearance of CI symptoms. PG activity diminished proportionally to the concentration of polyamine except for the concentration at 4.0 mM. No significant changes were observed in ethylene production.

Phenylalanine ammonia-lyase as related to ethylene in the development of chilling symptoms during cold storage of citrus fruits.

Low-temperature, nonfreezing, storage induces pitting and necrosis in the flavedo tissue of chilling susceptible citrus fruits. In this study the role of ethylene and phenylalanine ammonia-lyase (PAL; EC 4.3.1.5) in the cold-induced citrus peel damage has been investigated. It has been shown that increasing PAL activity by applying ethylene at a nonchilling temperature did not cause fruit damage or reduce the incidence of this peel disorder when fruits were subsequently held at a chilling temperature (2 degrees C). The cold-induced peel damage was enhanced by applying inhibitors of PAL activity and ethylene synthesis and action. These results indicate that the induction of PAL and ethylene during fruit cold storage, but not before, plays a role in reducing the development of chilling symptoms. The cold-induced PAL activity was reduced by inhibitors of ethylene production, but inhibitors of ethylene action exerted little effect on the activation of this enzyme. Therefore, the activation of PAL may be dependent on ethylene but also an independent cold signal apparently related to the cold-induced peel damage.

Effect of temperature on enzymatic and physiological factors related to chilling injury in carambola fruit (Averrhoa carambola L.).

Three groups of carambola fruits (Averrhoa carambola L.) were stored at 2 and 10 degrees C (85-90% relative humidity). The major physicochemical, physiological, and enzymatic responses of fruit were measured in each group over a 30-day period: chilling injury index (CII), decay (%), intracuticular waxes, cuticle permeability, pulp firmness, weight loss, sucrose, fructose and glucose contents, ion electrolyte leakage in pulp (%), ethylene and carbon dioxide production rates, and the activities of peroxidase (POD), polyphenol oxidase (PPO), and phenylalanine ammonia-lyase (PAL) enzymes. CII values were statistically different at 2 and 10 degrees C, showing high significance with respect to sucrose content and weight loss (P < 0.05). Chilling injury included darkened ribs and skin desiccation. According to the CI symptom development, a possible relationship of POD and PPO activities was found at 2 degrees C. A significant sucrose content increase was observed at 10 degrees C. CI symptoms were associated with POD and PAL activities.

[The relationship of pulp and periodontal disease in patients in a specialty clinic]. / Estudio sobre la relación de las enfermedades pulpares y periodontales en los pacientes de la clínica de especialidad.

This paper describes an epidemiological-clinical study performed at the Iztacala School of Professional Studies, a dependency of the National Autonomous University of Mexico. Records of 2,733 patients treated over a four year period were analyzed so as to examine the relationship between pulp and periodontal diseases, both of which are extremely prevalent in the Mexican population. On the basis of results thus yielded, it was possible to make specific recommendations regarding dental care following diagnosis.