The glucose transporter 4 FQQI motif is necessary for Akt Substrate of 160-Kilodalton-dependent plasma membrane translocation but not Golgi-localized g-ear-containing Arf-binding protein-dependent entry into the insulin-responsive storage compartment.

Newly synthesized glucose transporter 4 (GLUT4) enters into the insulin-responsive storage compartment in a process that is Golgi-localized γ-ear-containing Arf-binding protein (GGA) dependent, whereas insulin-stimulated translocation is regulated by Akt substrate of 160 kDa (AS160). In the present study, using a variety of GLUT4/GLUT1 chimeras, we have analyzed the specific motifs of GLUT4 that are important for GGA and AS160 regulation of GLUT4 trafficking. Substitution of the amino terminus and the large intracellular loop of GLUT4 into GLUT1 (chimera 1-441) fully recapitulated the basal state retention, insulin-stimulated translocation, and GGA and AS160 sensitivity of wild-type GLUT4 (GLUT4-WT). GLUT4 point mutation (GLUT4-F5A) resulted in loss of GLUT4 intracellular retention in the basal state when coexpressed with both wild-type GGA and AS160. Nevertheless, similar to GLUT4-WT, the insulin-stimulated plasma membrane localization of GLUT4-F5A was significantly inhibited by coexpression of dominant-interfering GGA. In addition, coexpression with a dominant-interfering AS160 (AS160-4P) abolished insulin-stimulated GLUT4-WT but not GLUT4-F5A translocation. GLUT4 endocytosis and intracellular sequestration also required both the amino terminus and large cytoplasmic loop of GLUT4. Furthermore, both the FQQI and the SLL motifs participate in the initial endocytosis from the plasma membrane; however, once internalized, unlike the FQQI motif, the SLL motif is not responsible for intracellular recycling of GLUT4 back to the specialized compartment. Together, we have demonstrated that the FQQI motif within the amino terminus of GLUT4 is essential for GLUT4 endocytosis and AS160-dependent intracellular retention but not for the GGA-dependent sorting of GLUT4 into the insulin-responsive storage compartment.

Glycogen storage capacity and de novo lipogenesis during massive carbohydrate overfeeding in man.

The metabolic balance method was performed on three men to investigate the fate of large excesses of carbohydrate. Glycogen stores, which were first depleted by diet (3 d, 8.35 +/- 0.27 MJ [1994 +/- 65 kcal] decreasing to 5.70 +/- 1.03 MJ [1361 +/- 247 kcal], 15% protein, 75% fat, 10% carbohydrate) and exercise, were repleted during 7 d carbohydrate overfeeding (11% protein, 3% fat, and 86% carbohydrate) providing 15.25 +/- 1.10 MJ (3642 +/- 263 kcal) on the first day, increasing progressively to 20.64 +/- 1.30 MJ (4930 +/- 311 kcal) on the last day of overfeeding. Glycogen depletion was again accomplished with 2 d of carbohydrate restriction (2.52 MJ/d [602 kcal/d], 85% protein, and 15% fat). Glycogen storage capacity in man is approximately 15 g/kg body weight and can accommodate a gain of approximately 500 g before net lipid synthesis contributes to increasing body fat mass. When the glycogen stores are saturated, massive intakes of carbohydrate are disposed of by high carbohydrate-oxidation rates and substantial de novo lipid synthesis (150 g lipid/d using approximately 475 g CHO/d) without postabsorptive hyperglycemia.

Audit report on the Iowa Petroleum Underground Storage Tank Board for the year ended June 30, 2013

Audit report on the Iowa Petroleum Underground Storage Tank Board for the year ended June 30, 2013

Reduced tillage: influence on erosion and nutrient losses in a clayey field in southern Finland

Selostus: Aurattoman viljelyn vaikutus eroosioon ja ravinnehuuhtoumiin eteläsuomlaisella, savimaalla sijaitsevalla pellolla

Ripening and quality of 'Laetitia' plums following harvest and cold storage as affected by inhibition of ethylene action

The inhibition of ethylene action by 1-methylcyclopropene (1-MCP) extends shelf and storage life of many climacteric fruits. However, 1-MCP appears to have limited effects on stone fruit depending on specie and cultivar. The effects of 1-MCP on ripening and quality of 'Laetitia' plums were determined during ripening at 23ºC following harvest and cold storage. Japanese plums (Prunus salicina, cv. Laetitia) were harvested at mature pre-climacteric stage, cooled to 2ºC within 36 hours of harvest and then treated with 0, 0.05, 0.10, 0.50 or 1.00 muL L-1 of 1-MCP at 1°C for 24 hours. Following treatment, fruits were either held at 23ºC for 16 days or stored at 1ºC for 50 days. Fruits were removed from cold storage at 10-day intervals and allowed to ripe at 23°C for five days. A delay of climacteric respiration and ethylene production by 1-MCP treatment during ripening following harvest and cold storage was associated to a slow rate of fruit softening. 1-MCP treatment also delayed the loss of titratable acidity and changes of flesh and skin color, whereas it had little or no effect on soluble solids content. 1-MCP effects were concentration- and storage duration-dependent and, generally, a saturation fruit response to 1-MCP occurred between 0.5 and 1.0 muL L-1. During ripening, 1-MCP treated fruits attained quality similar to that of controls. Results indicated that 1-MCP treatment may extend shelf life (23ºC) and storage life (1ºC) of 'Laetitia' plums by approximately six and 20 days, respectively.

Use of antimicrobial biodegradable packaging to control Listeria monocytogenes during storage of cooked ham

The antimicrobial effect against L. monocytogenes of biodegradable films (alginate, zein and polyvinyl alcohol) containing enterocins was investigated. Survival of the pathogen was studied by means of challenge tests performed at 6 °C during 8 and 29 days, for air-packed and vacuum-packed sliced cooked ham, respectively. Air packaging was tested with two concentrations of enterocins (200 and 2000 AU/cm2). Control air-packed cooked ham showed an increase of L. monocytogenes from 104 to 107 CFU/g after 8 days. By contrast, packaging with antimicrobial films effectively slowed down the pathogen's growth, leading to final counts lower than in control lots. Air-packaging with alginate films containing 2000 AU/cm2 of enterocins effectively controlled L. monocytogenes for 8 days. An increase of only 1 log unit was observed in zein and polyvinyl alcohol lots at the same enterocin concentration. Vacuum packaging with films containing enterocins (2000 AU/cm2) also delayed the growth of the pathogen. No increase from inoculated levels was observed during 15 days in antimicrobial alginate films. After 29 days of storage, the lowest counts were obtained in samples packed with zein and alginate films containing enterocins, as well as with zein control films. The most effective treatment for controlling L. monocytogenes during 6 °C storage was vacuum-packaging of sliced cooked ham with alginate films containing 2000 AU/cm2 of enterocins. From the results obtained it can concluded that antimicrobial packaging can improve the safety of sliced cooked ham by delaying and reducing the growth of L. monocytogenes.

High-pressure processing and antimicrobial biodegradable packaging to control Listeria monocytogenes during storage of cooked ham

The efficiency of combining high-pressure processing (HPP) and active packaging technologies to control Listeria monocytogenes growth during the shelf life of artificially inoculated cooked ham was assessed. Three lots of cooked ham were prepared: control, packaging with alginate films, and packaging with antimicrobial alginate films containing enterocins. After packaging, half of the samples were pressurized. Sliced cooked ham stored at 6 °C experienced a quick growth of L. monocytogenes. Both antimicrobial packaging and pressurization delayed the growth of the pathogen. However, at 6 °C the combination of antimicrobial packaging and HPP was necessary to achieve a reduction of inoculated levels without recovery during 60 days of storage. Further storage at 6 °C of pressurized antimicrobial packed cooked ham resulted in L. monocytogenes levels below the detection limit (day 90). On the other hand, storage at 1 °C controlled the growth of the pathogen until day 39 in non-pressurized ham, while antimicrobial packaging and storage at 1 °C exerted a bacteriostatic effect for 60 days. All HPP lots stored at 1 °C led to counts <100 CFU/g at day 60. Similar results were observed when combining both technologies. After a cold chain break no growth of L. monocytogenes was observed in pressurized ham packed with antimicrobial films, showing the efficiency of combining both technologies.

Combined effect of natural antimicrobials and high pressure processing to prevent Listeria monocytogenes growth after a cold chain break during storage of cooked ham

The effect of high pressure processing (400 MPa for 10 min) and natural antimicrobials 2 (enterocins and lactate-diacetate) on the behaviour of L. monocytogenes in sliced cooked ham 3 during refrigerated storage (1ºC and 6ºC) was assessed. The efficiency of the treatments after a 4 cold chain break was evaluated. Lactate-diacetate exerted a bacteriostatic effect against L. 5 monocytogenes during the whole storage period (3 months) at 1ºC and 6ºC, even after 6 temperature abuse. The combination of low storage temperature (1ºC), high pressure 7 processing (HPP) and addition of lactate-diacetate reduced the levels of L. monocytogenes 8 during storage by 2.7 log CFU/g. The most effective treatment was the combination of HPP, 9 enterocins and refrigeration at 1ºC, which reduced the population of the pathogen to final counts 10 of 4 MPN/g after 3 months of storage, even after the cold chain break.

Emergence, longevity and fecundity of Trissolcus basalis and Telenomus podisi after cold storage in the pupal stage

Pupae of Trissolcus basalis (Wollaston) and Telenomus podisi Ashmead (Hymenoptera: Scelionidae) were stored at 12ºC and 15ºC for 120-210 days, after different periods of parasitism at 18ºC in order to evaluate adult emergence, longevity and ovipositional capacity. There was no emergence of adults at 12ºC. The rate of emergence of parasitoids transferred to 15ºC at the beginning of the pupal stage was 1.5% and 26.3%, for T. basalis and T. podisi respectively, whereas those parasitoids transferred one day before the expected date of emergence at 18ºC showed 86.4% of emergence for T. basalis and 59.9% for T. podisi. Mean adult longevity was also significantly lower when pupae were transferred to 15ºC at the beginning of the pupal stage. Females emerged after storage and maintained for 120 to 210 days at 15ºC parasitized host eggs after transference to 25ºC; however, fecundity of T. podisi was reduced in about 80% after cold storage.

Basil conservation affected by cropping season, harvest time and storage period

Fresh basil (Ocimum basilicum L.) is used in food, phytotherapic industry, and in traditional therapeutic, due to its essential oil content and composition. Nevertheless basil can not be kept for long periods after harvest and its quality can be reduced. This work aimed to assess the influence of the season and harvest time in the postharvest conservation of basil stored for different periods. Basil was harvested at 8 am and 4 pm both in August/1999 and January/2000. Cuttings were conditioned in PVC packages and stored for 3, 6, and 9 days. During storage, chlorophyll content, essential oil content and composition were determined as well as microbiological analyses were carried out. Harvest season and the days of storage influenced the final content of essential oil. There was a linear decrease in the content of essential oil, in the chlorophyll content and in the number of mold and yeast colonies during storage. There was no effect of cropping season or harvest hour on essential oil composition, but the eugenol and linalool content increased during storage. Coliforms were under 0.3 MPN g-1 and the number of Staphylococcus aureus was under 1.0x10² UFC g-1.

Conservation of 'Pedro Sato' guavas under treatment with 1-methylcyclopropene

The search for techniques that extend shelf life of guava (Psidium guajava) fruits, and reduce its postharvest losses is desirable. The objective of this work was to evaluate the effects of concentrations of competitive ethylene antagonist 1-methylcyclopropene (1-MCP) on conservation of 'Pedro Sato' guava fruits. Treatments consisted of 0, 100, 300 or 900 nL L-1 of 1-MCP for 3 hours followed by storage at 25ºC, and 10ºC with 90±5% RH. The application of 900 nL L-1 1-MCP for 3 hours was efficient in delaying loss of skin color, and in keeping fruit firmness at both storage temperatures. The 300 nL L-1 of 1-MCP concentration was efficient in delaying skin color loss only when fruits were stored at 10ºC. The effect of 1-MCP was quite significant on the reduction of rot incidence at both storage temperatures. The respiratory rate was lower in fruits treated with 300 and 900 nL L-1 of 1-MCP during storage at 25ºC. The product was efficient in delaying the ripening of fruits, and the concentration of 900 nL L-1 showed the best effect.

Storage and uptake of D-serine into astrocytic synaptic-like vesicles specify gliotransmission.

Glial cells are increasingly recognized as active players that profoundly influence neuronal synaptic transmission by specialized signaling pathways. In particular, astrocytes have been shown recently to release small molecules, such as the amino acids l-glutamate and d-serine as "gliotransmitters," which directly control the efficacy of adjacent synapses. However, it is still controversial whether gliotransmitters are released from a cytosolic pool or by Ca(2+)-dependent exocytosis from secretory vesicles, i.e., by a mechanism similar to the release of synaptic vesicles in synapses. Here we report that rat cortical astrocytes contain storage vesicles that display morphological and biochemical features similar to neuronal synaptic vesicles. These vesicles share some, but not all, membrane proteins with synaptic vesicles, including the SNARE (soluble N-ethylmaleimide-sensitive factor attachment protein receptor) synaptobrevin 2, and contain both l-glutamate and d-serine. Furthermore, they show uptake of l-glutamate and d-serine that is driven by a proton electrochemical gradient. d-Serine uptake is associated with vesicle acidification and is dependent on chloride. Whereas l-serine is not transported, serine racemase, the synthesizing enzyme for d-serine, is anchored to the membrane of the vesicles, allowing local generation of d-serine. Finally, we reveal a previously unexpected mutual vesicular synergy between d-serine and l-glutamate filling in glia vesicles. We conclude that astrocytes contain vesicles capable of storing and releasing d-serine, l-glutamate, and most likely other neuromodulators in an activity-dependent manner.

Electrical conductivity testing of corn seeds as influenced by temperature and period of storage

The objective of this work was to evaluate the effects of temperature (10, 20, 30, 20/10 and 30/10ºC) and period of storage on electrical conductivity (EC) in four seed lots of corn (Zea mays L.), as well as the mineral composition of the soaking solution. EC test determines indirectly the integrity of seed membrane systems, and is used for the assessment of seed vigor, because this test detects the seed deterioration process since its early phase. The research comprised determinations of water content, germination, accelerated aging (AA), cold (CT) and EC vigor tests, and determinations of Ca2+, Mg2+ and K+ release to the solution, after seed soaking of four corn seed lots. The evaluations were performed each four months during a period of 16 months. For statistical analysis, a completely randomized split plot design was used with eight replications. Except for seed lots stored at 10ºC, all vigor evaluations revealed a decline in vigor, but AA and CT showed more sensitiveness to declines of seed physiological quality than EC. Potassium was the main leached ion regardless of the storage temperature.

Postharvest quality of peaches harvested from integrated and conventional production systems

Over three years the postharvest quality of 'Marli' peaches harvested from the integrated (IFP) and conventional production (CFP) systems was evaluated. The peaches were harvested from commercial orchards of Prunus persica at two locations close to the city of São Jerônimo, RS, Brazil, and stored at 0.5°C for 10, 20 or 30 days. The peaches were evaluated at harvest, at retrieval from storage and after ripening periods at 20°C. No differences in fruit weight losses were determined. Decay incidence was low, and no differences were detected amongst systems in both 2001 and 2002 seasons, but in the 2000 season CFP peaches were more decayed. Flesh firmness of peaches from the IFP system were greater than CFP fruits in the years 2000 and 2001. In 2002, firmness changed little during storage and ripening. Peaches from the IFP in 2000 had higher titratable acidity and lower soluble solids. In the 2000 season, flesh browning was observed in decayed fruits, always at ripening after 20 or 30 days of cold storage Chilling injuries such as browning, woolliness and leatheriness ocurred in 2002. There were no differences amongst systems related to peach quality.