Intracellular targeting of GLUT4 in transfected insulinoma cells: evidence for association with constitutively recycling vesicles distinct from synaptophysin and insulin vesicles.

In adipocytes and muscle cells, the GLUT4 glucose transporter isoform is present in intracellular vesicles which continuously recycle between an intracytoplasmic location and the plasma membrane. It is not clear whether the GLUT4-vesicles represent a specific kind of vesicle or resemble typical secretory granules or synaptic-like microvesicles. To approach this question, we expressed GLUT4 in the beta cell line RINm5F and determined its intracellular localization by subcellular fractionation and by immunofluorescence and immunoelectron microscopy. GLUT4 was not found in insulin granules but was associated with a subpopulation of smooth-surface vesicles present in the trans-Golgi region and in vesicular structures adjacent to the plasma membrane. In the trans-Golgi region, GLUT4 did not colocalize with synaptophysin or TGN38. Incubation of the cells with horseradish peroxidase (HRP) led to colocalization of HRP and GLUT4 in some endosomal structures adjacent to the plasma membrane and in occasional trans-Golgi region vesicles. When cells were incubated in the presence of Bafilomycin A, analysis by confocal microscopy revealed GLUT4 in numerous large spots present throughout the cytoplasm, many of which costained for TGN38 and synaptophysin. By immunoelectron microscopy, numerous endosomes were observed which stained strongly for GLUT4. Together our data demonstrate that ectopic expression of GLUT4 in insulinoma cells reveals the presence of a subset of vesicular structures distinct from synaptic-like vesicles and insulin secretory granules. Furthermore, they indicate that GLUT4 constitutively recycles between the plasma membrane and its intracellular location by an endocytic route also taken by TGN38 and synaptophysin.

Elektroniikka - ja sähköromun (SER) kierrätys

Tässä työssä pyrin tarkastelemaan sähkö- ja elektroniikkaromun kierrätystä erityisesti EU - direktiivin mukaisen tuottajavastuun näkökulmasta. Käytettyjen sähkö- ja elektroniikkalaitteiden vuori kasvaa koko ajan kaikkialla Euroopassa. Euroopan parlamentti hyväksyi joulukuussa 2002 kaksi uutta direktiiviä, joilla pyritään lieventämään sähköromuongelmaa. Parlamentin vaatimusten ansiosta unionin jäsenvaltioiden on taattava, että laitteiden valmistajat maksavat omista tuotteistaan syntyvän sähkö- ja elektroniikkalaitejätteen keräys-, käsittely- ja kierrätyskustannukset. Tämä merkitsee esimerkiksi sitä, että kuluttajat voivat viedä vanhat sähkölaitteensa asuinpaikkansa läheisyydessä sijaitseviin maksuttomiin keräyspisteisiin. Myös kuluttajien on otettava vastuu hankkimistaan tuotteista: sähkö- ja elektroniikkalaitejätteen hävittäminen tavallisen kotitalousjätteen seassa tullaan kieltämään.

Polarized cell growth in Arabidopsis requires endosomal recycling mediated by GBF1-related ARF exchange factors.

Polarized tip growth is a fundamental cellular process in many eukaryotic organisms, mediating growth of neuronal axons and dendrites or fungal hyphae. In plants, pollen and root hairs are cellular model systems for analysing tip growth. Cell growth depends on membrane traffic. The regulation of this membrane traffic is largely unknown for tip-growing cells, in contrast to cells exhibiting intercalary growth. Here we show that in Arabidopsis, GBF1-related exchange factors for the ARF GTPases (ARF GEFs) GNOM and GNL2 play essential roles in polar tip growth of root hairs and pollen, respectively. When expressed from the same promoter, GNL2 (in contrast to the early-secretory ARF GEF GNL1) is able to replace GNOM in polar recycling of the auxin efflux regulator PIN1 from endosomes to the basal plasma membrane in non-tip growing cells. Thus, polar recycling facilitates polar tip growth, and GNL2 seems to have evolved to meet the specific requirement of fast-growing pollen in higher plants.

KIAA1985, a Protein Mutant in Charcot-Marie-Tooth Neuropathy, Links Peripheral Nerve Myelination to Endosomal Recycling Pathways

Charcot-Marie-Tooth neuropathy (CMT) represents a heterogenous group of inherited disorders of the peripheral nervous system. One form of autosomal recessive demyelinating CMT (CMT4C, 5q32) is caused by mutations in the gene encoding KIAA1985, a protein of so far unknown function. Here we show that KIAA1985 is exclusively expressed in Schwann cells. KIAA1985 is tethered to cellular membranes through an N-terminal myristic acid anchor and localizes to the perinuclear recycling compartment. A search for proteins that interact with KIAA1985 identified the small GTPase Rab11, a key regulator of recycling endosome functions. CMT4C-related missense mutations disrupt the KIAA1985/Rab11 interaction. Protein binding studies indicate that KIAA1985 functions as a Rab11 effector, as it interacts only with active forms of Rab11 (WT and Q70L) and does not interact with the GDP locked mutant (S25N). Consistent with a function of Rab11 in Schwann cell myelination, myelin formation was strongly impaired when dorsal root ganglion neurons were co-cultured with Schwann cells infected with Rab11 S25N. Our data indicate that the KIAA1985/Rab11 interaction is relevant for peripheral nerve pathophysiology and place endosomal recycling on the list of cellular mechanisms involved in Schwann cell myelination.

A modular tethering complex for endosomal recycling.

How proteins migrate through the interconnected organelles of the endolysosomal system is poorly understood. A piece of the puzzle has been added with the identification of a complex of tethering factors that functions in the recycling of proteins towards the cell surface.

Biochemical treatment of brown grade rejects in recycling process

Ruskeisiin kierrätysmassoihin kuuluu kulutuksen kannalta tärkeimpänä laatuna OCC (old corrugated containers). OCC sisältää noin 70-100% aaltopahvia eli pääasiassa se koostuu valkaisemattomasta kemiallisesta massasta. OCC uusiomassan ensisijainen käyttökohde on aaltopahvin valmistus. OCC:n kierrätyskuituprosessissa syntyy merkittäviä määriä rejektiä. Rejektin määrä riippuu paljolti kierrätettävän materiaalin laadusta ja puhtaudesta, mutta myös tulevan massan käyttötarkoituksesta sekä prosessiolosuhteista. OCC-prosessissa rejektoituvan aineksen määrä voi nousta korkeaksi, mikäli kierrätettävä materiaali sisältää märkälujaliimoja tai muuten raskaasti liimattuja komponentteja sekä runsaasti kontaminantteja, kuten muoveja, teippejä ja metalleja. Keskimäärin OCC-rejekti sisältää 30-60% kiinteää ainesta, 30-90% (kuivapaino) kuituja, 5-70% (kuivapaino) muoveja ja 1-10% (kuivapaino) tuhkaa. Syntynyt rejekti voidaan polttaa energiaksi tai käyttää maantäyttöaineena. Harvinaisempia sovelluksia rejektin käsittelyssä ovat rejektin kuitujen talteenotto uudelleenprosessointia varten tai alkoholin ja levuliinihapon tuottamiseen. Rejektin asianmukaisella käsittelyllä voidaan vähentää kaatopaikkakustannuksia, sekä parantaa kierrätysprosessin tuottavuutta. Tämän työn tarkoituksena oli tutkia biokemiallisen käsittelyn mahdollisuudet OCC-rejektin hajotuksessa. Alustavissa laboratoriomittakaavan kokeissa etsittiin sopiva käsittelytapa, joka toteutettiin sitten pilot plant -mittakaavassa. Tulokset osoittavat, että biokemiallisen käsittelyn avulla rejekti voidaan hajottaa jolloin jätteenkäsittelykustannukset pienenevät ja kierrätyskuituprosessin taloudellisuus paranee.

Industry Change in Transport Packaging Sector. Case: Eltete TPM Ltd

Tämän tutkielman tarkoituksena on selvittää suomalaisen kartonkikuljetuspakkausfirman, Eltete TPM Oy:n, tulevaisuudennäkymät selvittämällä voiko puisen kuljetuspakkausmateriaalin korvata kartonkisilla materiaaleilla. Kyseinen tutkimus suoritetaan selvittämällä toimialanmuutokseen vaikuttavia tekijöitä kuten ostokäyttäytymistä, innovaation ominaisuuksia, säädöksiä, kilpailuympäristöä sekä lopuksi tutkimalla mitä kuljetuspakkauksien asiakkaat haluavat ja tarvitsevat. Kyseisen tutkimuksen kohdeasiakkaiksi valittiin suurimmat kodinkoneyritykset Euroopassa ja tutkimusmenetelmäksi postikysely. Suoritetun markkinointitutkimuksen tulokset kodinkoneyritysten pakkausratkaisuista sekä kilpailija-analyysi osoittivat, että todella on olemassa kasvava kysyntä kierrätettäville kartonkisille kuljetuspakkausratkaisuille, kuten Eltete TPS - Framepack solution on, sekä että Eltete TPM Oy:llä on voimavaroja tulevaisuudessa nousta markkinajohtajaksi muuttuvalla kuljetuspakkaussektorilla.

Effects of unit - based pricing on the waste collection demand : a meta - regression analysis

We perform a meta - analysis of 21 studies that estimate the elasticity of the price of waste collection demand upon waste quantities, a prior literature review having revealed that the price elasticity differs markedly. Based on a meta - regression with a total of 65 observations, we find no indication that municipal data give higher estimates for price elasticities than those associated with household data. Furthermore, there is no evidence that treating prices as exogenous underestimates the price elasticity. We find that much of the variation can be explained by sample size, the use of a weight - based as opposed to a volume - based pricing system, and the pricing of compostable waste. We also show that price elasticities determined in the USA and point estimations of elasticities are more elastic, but these effects are not robust to the changing of model specifications. Finally, our tests show that there is no evidence of publication bias while there is some evidence of the existence of genuine empirical effect.

Sulfuric acid - glucose separation using mixed-recycle steady state recycling chromatography

Batch chromatography is a widely used separation technique in a variety of fields meeting difficult separations. Several technologies for improving the performance of chromatography have been studied, including mixed-recycle steady state recycling (MR-SSR) chromatography. Design of MR-SSR has been commonly limited on 100 % purity constraint cases and empirical work. In this study a predictive design method was used to optimize feed pulse size and design a number of experimental MR-SSR separations for a solution of 20 % sulfuric acid and 100 g/L glucose. The design was under target product fraction purities of 98.7 % for H2SO4 and 95 % for glucose. The experiments indicate a maximum of 59 % increase in sulfuric acid productivity and 82 % increase for glucose when compared to corresponding batch separation. Eluent consumption was lowered by approximately 50 % using recycling chromatography. Within this study the target purities and yields set in design were not completely met, and further optimization of the process is deemed necessary.

Recycling of nutrients with application of organic waste in degraded pasture

The utilization of organic wastes represents an alternative to recover degraded pasture. The experiment aimed to assess the changes caused by the provision of different organic waste (poultry litter, turkey litter and pig manure) in a medium-textured Oxisol in Brazilian Savanna under degraded pasture. It was applied different doses of waste compared to the use of mineral fertilizers and organic mineral and evaluated the effect on soil parameters (pH, organic matter, phosphorus and potassium) and leaf of Brachiariadecumbens (crude protein, phosphorus and dry mass production). It was observed that application of organic waste did not increase the level of soil organic matter and pH in the surface layer, and the application of turkey litter caused acidification at depths of 0.20-0.40 m and 0.40-0.60 m. There was an increase in P and K in the soil with the application of poultry litter and swine manure. All organic wastes increased the productivity of dry matter and crude protein and phosphorus. The recycling of nutrients via the application of organic waste allows efficiency of most parameters similar to those observed with the use of mineral sources, contributing to improving the nutritional status of soil-plantsystem.

Factors associated with fragility fractures in women over 50 years of age: a population-based household survey

PURPOSE: To analyze the prevalence of and factors associated with fragility fractures in Brazilian women aged 50 years and older. METHODS: This cross-sectional population survey, conducted between May 10 and October 31, 2011, included 622 women aged >50 years living in a city in southeastern Brazil. A questionnaire was administered to each woman by a trained interviewer. The associations between the occurrence of a fragility fracture after age 50 years and sociodemographic data, health-related habits and problems, self-perception of health and evaluation of functional capacity were determined by the χ2 test and Poisson regression using the backward selection criteria. RESULTS: The mean age of the 622 women was 64.1 years. The prevalence of fragility fractures was 10.8%, with 1.8% reporting hip fracture. In the final statistical model, a longer time since menopause (PR 1.03; 95%CI 1.01-1.05; p<0.01) and osteoporosis (PR 1.97; 95%CI 1.27-3.08; p<0.01) were associated with a higher prevalence of fractures. CONCLUSIONS: These findings may provide a better understanding of the risk factors associated with fragility fractures in Brazilian women and emphasize the importance of performing bone densitometry.

Decentralized management of organic household wastes in the Kathmandu Valley using small-scale composting reactors

The sustainable management of municipal solid waste in the Kathmandu Valley has always been a challenging task. Solid waste generation has gone rapidly high in the Kathmandu Valley over the last decade due to booming population and rapid urbaniza-tion. Finding appropriate landfill sites for the disposal of solid wastes generated from the households of the Kathmandu Valley has always been a major problem for Nepalese government. 65 % of total generated wastes from the households of Nepal consist of organic materials. As large fractions of generated household wastes are organic in na-ture, composting can be considered as one of the best sustainable ways to recycle organ-ic wastes generated from the households of Nepal. Model Community Society Development (MCDS), a non-governmental organization of Nepal carried out its small-scale project in five households of the Kathmandu Valley by installing composting reactors. This thesis is based on this small-scale project and has used secondary data provided by MCDS Nepal for carrying out the study. Proper man-agement of organic wastes can be done at household levels through the use of compost-ing reactors. The end product compost can be used as soil conditioners for agricultural purposes such as organic farming, roof-top farming and gardening. The overall average organic waste generation in the Kathmandu Valley is found to be 0,23 kg/person/day and the total amount of organic household wastes generated in the Kathmandu Valley is around 210 Gg/yr. Produced composts from five composting reac-tors contain high amount of moistures but have sufficient amount of nutrients required for the fertility of land and plant growth. Installation of five composting reactors in five households have prevented 2,74 Mg of organic wastes going into the landfills, thus re-ducing 107 kg of methane emissions which is equivalent to 2,7 Mg of carbondioxide.

Yhdyskuntajätteen mekaanisen erottelun erotustehokkuus

EU:n jätehierarkia asettaa jätteenkäsittelyssä materiaalien hyötykäytön energiahyötykäytön edelle. EU on asettanut korkeat tavoitteet jätteenkierrätykseen, 50 painoprosenttia kotitalousjätteestä on ohjattava kierrätykseen vuoteen 2020 mennessä. Suomessa kaatopaikoista on pyritty eroon lisäämällä jätteenpolttokapasiteettia. Jätteiden hyödyntämisen osalta tilanne Suomessa on hyvä, mutta kierrätystavoitteiden täyttyminen nykyisillä toimilla vaikuttaa epätodennäköiseltä. Tässä työssä selvitetään mitä mekaanisia jätteen erottelumenetelmiä maailmalla on käytössä ja kuinka tehokkaita ne ovat. Työn tavoitteena on tutkia voitaisiinko kierrätystä Suomessa tehostaa yhdyskuntajätteen mekaanisella käsittelyllä. Kirjallisuusselvityksen lisäksi työssä on simuloitu mekaanisia erotteluketjuja ja verrattu niillä saatuja tuloksia Suomen syntypaikkalajittelun tasoon. Tämän tutkimuksen perusteella, mikään yksittäinen mekaaninen erottelumenetelmä ei riittävän tehokas erottelemaan kierrätettäviä materiaaleja yhdyskuntajätteestä. Mekaanisia erottelumenetelmiä tulee yhdistää lajittelulinjastoiksi, joiden optimoiminen on monen tekijän summa. Lajittelulinjaston suunnitteluun vaikuttavat muun muassa lähtömateriaalin laatu ja lopputuotteiden käyttötarkoitukset. Yhdyskuntajätteen sisältämä biojäte likaa herkästi muut jätteet ja vaikeuttaa mekaanisesti eroteltujen jätejakeiden uudelleenkäyttöä. Biojätteen poistaminen muiden jätteiden joukosta olisi ensiarvoisen tärkeää mekaanisen erotuksen tehokkuuden kannalta. Mekaaniset erotteluketjut poistavat tehokkaasti biojätettä ja metalleja, mutta lasin ja kuitujen osalta erotusketjujen tehokkuudet jäävät alhaisiksi. Muovien osalta mekaaninen erottelu voi parhaimmillaan ollaan erittäin tehokasta, toisaalta vaatimukset lähtömateriaalin laadulle ovat suuret. Muovien osalta syntypaikkalajittelun ja mekaanisen erottelun yhtäaikainen tehostaminen voisi tarjota ratkaisun kierrätysasteen nostamiseen.

Effect of Material Recycling on the Feasibility of Solid Recovered Fuel Fired Power Plant

This master’s thesis examines the effects of increased material recycling on different waste-to-energy concepts. With background study and a developed techno-economic computational method the feasibility of chosen scenarios with different combinations of mechanical treatment and waste firing technologies can be evaluated. The background study covers the waste scene of Finland, and potential market areas Poland and France. Calculated cases concentrate on municipal solid waste treatment in the Finnish operational environment. The chosen methodology to approach the objectives is techno-economic feasibility assessment. It combines calculation methods of literature and practical engineering to define the material and energy balances in chosen scenarios. The calculation results together with other operational and financial data can be concluded to net present values compared between the scenarios. For the comparison, four scenarios, most vital and alternative between each other, are established. The baseline scenario is grate firing of source separated mixed municipal solid waste. Second scenario is fluidized bed combustion of solid recovered fuel produced in mechanical treatment process with metal separation. Third scenario combines a biomaterial separation process to the solid recovered fuels preparation and in the last scenario plastics are separated in addition to the previous operations. The results indicated that the mechanical treatment scenarios still need to overcome some problems to become feasible. Problems are related to profitability, residue disposal and technical reliability. Many uncertainties are also related to the data gathered over waste characteristics, technical performance and markets. With legislative support and development of further processing technologies and markets of the recycled materials the scenarios with biomaterial and plastic separation may operate feasibly in the future.