Aging preferentially affects molecular pathways implicated in development and disease of myelinating glial cells

The central and peripheral nervous systems are involved in multiple age-dependent neurological deficits that are often attributed to alterations in function of myelinating glial cells. However, the molecular events that underlie the age-related decline of glial cell function are unknown. We used Schwann cells as a model to study biological processes affected in glial cells by aging. We comprehensively profiled gene expression of the Schwann cellrich mouse sciatic nerve throughout life, from day of birth until senescence (840 days of age). We combined the aging data with the microarray transcriptional data obtained using nerves isolated from Schwann cell-specific neuropathy-inducing mutants MPZCre/+/Lpin1fE2−3/fE2−3 , MPZCre/+/ScapfE1/fE1 and Pmp22-null mice. The majority of age related transcripts were also affected in the analyzed mouse models of neuropathy (54.4%) and in development (59.5%) indicating a high level of overlapping in implicated molecular pathways. We observed that compared to peripheral nerve development, dynamically changing expression profiles in aging have opposite (anticorrelated) orientation while they copy the orientation of transcriptional changes observed in analyzed neuropathy models. Subsequent clustering and biological annotation of dynamically changing transcripts revealed that the processes most significantly deregulated in aging include inflammatory/immune response and lipid biosynthesis/metabolism. Importantly, the changes in these pathways were also observed in myelinated oligodendrocyte-rich optic nerves of aged mice, albeit with lower magnitude. This observation suggests that similar biological processes are affected in aging glial cells in central and peripheral nervous systems, however with different dynamics. Our data, which provide the first comprehensive comparison of molecular changes in glial cells in three distinct biological conditions comprising development, aging and disease, provide not only a new inside into the molecular alterations underlying neural system aging but also identify target pathways for potential therapeutic approaches to prevent or delay complications associated with age-related and inherited forms of neuropathies. *Current address: Department of Physiology, UCSF, San Francisco, CA, USA.

Galvanomagnetic effects in strongly doped p-Bi2Te3:Sn crystals

Magnetic field dependencies of Hall coefficient and magnetoresistivity are investigated in classical and quantizing magnetic fields in p-Bi2Te3 crystals heavily doped with Sn grown by Czochralsky method. Magnetic field was parallel to the trigonal axis C3. Shubnikov-de Haas effect and quantum oscillations of the Hall coefficient were measured at temperatures 4.2 K and 11 K. On the basis of the magnetic field dependence of the Hall coefficient a method of estimation of the Hall factor and Hall mobility using the Drabble- Wolf six ellipsoid model is proposed. Shubnikov-de Haas effect and quantum oscillations of the Hall coefficient were observed at 4.2 K and 11 K. New evidence for the existence of the narrow band of Sn impurity states was shown. This band is partly filled by electrons and it is overlapping with the valence states of the light holes. Parameters of the impurity states, their energy ESn - 15 meV, band broadening ¿<< k0T and localization radius of the impuritystate R - 30 Å were obtained.

Current Issues in the Diagnosis and Classification of Peripheral T-Cell Lymphomas

Mature T-cell and T/NK-cell neoplasms are both uncommon and heterogeneous, among the broad category of non-Hodgkin's lymphomas. Due to the lack of specific genetic alterations in the vast majority of cases, most currently defined entities show overlapping morphologic and immunophenotypic features and therefore pose a challenge to the diagnostic pathologist. The goal of the symposium is to address current criteria for the recognition of specific subtypes of T-cell lymphoma, and to highlight new data regarding emerging immunophenotypic or molecular markers. This activity has been designed to meet the needs of practicing pathologists, and residents and fellows enrolled in training programs in anatomic and clinical pathology. It should be a particular benefit to those with an interest in hematopathology. Upon completion of this activity, participants should be better able to: -To be able to state the basis for the classification of mature T-cell malignancies involving nodal and extranodal sites. -To recognize and accurately diagnose the various subtypes of nodal and extranodal peripheral T-cell lymphomas. -To utilize immunohistochemical and molecular tests to characterize atypical T-cell proliferations. -To recognize and accurately diagnose T-cell lymphoproliferative lesions involving the skin and gastrointestinal tract, and be able to provide guidance regarding their clinical aggressiveness and management -To be able to utilize flow cytometric data to identify diverse functional T-cell subsets.

Working memory load improves early stages of independent visual processing.

Increasing evidence suggests that working memory and perceptual processes are dynamically interrelated due to modulating activity in overlapping brain networks. However, the direct influence of working memory on the spatio-temporal brain dynamics of behaviorally relevant intervening information remains unclear. To investigate this issue, subjects performed a visual proximity grid perception task under three different visual-spatial working memory (VSWM) load conditions. VSWM load was manipulated by asking subjects to memorize the spatial locations of 6 or 3 disks. The grid was always presented between the encoding and recognition of the disk pattern. As a baseline condition, grid stimuli were presented without a VSWM context. VSWM load altered both perceptual performance and neural networks active during intervening grid encoding. Participants performed faster and more accurately on a challenging perceptual task under high VSWM load as compared to the low load and the baseline condition. Visual evoked potential (VEP) analyses identified changes in the configuration of the underlying sources in one particular period occurring 160-190 ms post-stimulus onset. Source analyses further showed an occipito-parietal down-regulation concurrent to the increased involvement of temporal and frontal resources in the high VSWM context. Together, these data suggest that cognitive control mechanisms supporting working memory may selectively enhance concurrent visual processing related to an independent goal. More broadly, our findings are in line with theoretical models implicating the engagement of frontal regions in synchronizing and optimizing mnemonic and perceptual resources towards multiple goals.

Regulation of the expression of Nav1.5, the cardiac voltage-gated sodium channel

Abstract The cardiac sodium channel Nav1.5 plays a key role in cardiac excitability and conduction. Its importance for normal cardiac function has been highlighted by descriptions of numerous mutations of SCN5A (the gene encoding Nav1.5), causing cardiac arrhythmias which can lead to sudden cardiac death. The general aim of my PhD research project has been to investigate the regulation of Nav1.5 along two main axes: (1) We obtained experimental evidence revealing an interaction between Nav1.5 and a multiprotein complex comprising dystrophin. The first part of this study reports the characterization of this interaction. (2) The second part of the study is dedicated to the regulation of the cardiac sodium channel by the mineralocorticoid hormone named aldosterone. (1) Early in this study, we showed that Nav1.5 C-terminus was associated with dystrophin and that this interaction was mediated by syntrophin proteins. We used dystrophin-deficient mdx5cv mice to study the role of this interaction. We reported that dystrophin deficiency led to a reduction of both Nav1.5 protein level and the sodium current (INa). We also found that mdx5cv mice displayed atrial and ventricular conduction defects. Our results also indicated that proteasome inhibitor MG132 treatment of mdx5cv mice rescued Nav1.5 protein level and INa in cardiac tissue. (2) We showed that aldosterone treatment of mice cardiomyocytes led to an increase of the sodium current with no modification of Nav1.5 transcript and protein level. Altogether, these results suggest that the sodium current can be increased by distribution of intracellular pools of protein to the plasma membrane (e.g. upon aldosterone stimulation) and that interaction with dystrophin multiprotein complex is required for the stabilization of the channel at the plasma membrane. Finally, we obtained preliminary results suggesting that the proteasome could regulate Nav1.5 in mdx5cv mice. This study defines regulatory mechanisms of Nav1.5 which could play an important role in cardiac arrhythmia and bring new insight in cardiac conduction alterations observed in patients with dystrophinopathies. Moreover, this work suggests that Brugada syndrome, and some of the cardiac alterations seen in Duchenne patients may be caused by overlapping molecular mechanisms leading to a reduction of the cardiac sodium current.

Deposición transversal de líquido de las boquillas de doble abanico TTJ60-11004 y TTJ60-11002 en distintas condiciones operacionales

El objetivo de este trabajo fue el de evaluar la deposición transversal de caldo de las boquillas pulverizadoras de doble abanico TTJ60-11004 y TTJ60-11002 en distintas condiciones operacionales. Se utilizaron 5 muestras de cada boquilla pulverizadora siendo considerada cada unidad, una repetición. La distribución de caldo fue evaluada por medio de una mesa de evaluación de distribución construida de acuerdo con la norma ISO 56821. Se evaluó el perfil de distribución individual, la distribución volumétrica simulada de la superposición de los chorros por medio del coeficiente de variación (CV%) de los volúmenes colectados, el caudal y el ángulo de abertura de los chorros. Las condiciones operacionales fueron: presión de trabajo de 200, 300 y 400 Kpa, altura de 30, 40 y 50 cm en relación al blanco y espaciamiento entre boquillas simulados en Software (Microsoft Excel) entre 45 y 100 cm. Las boquillas presentaron perfil individual descontinuo con la mayor deposición de líquido en la región central y reducción del volumen gradual en dirección a las extremidades. El aumento de la presión promovió alargamiento del perfil y de la franja de aplicación. Las boquillas proporcionaron perfil uniforme que dependió del espaciamiento entre las boquillas, con valores menores con reducción en el espaciamiento y en presiones mayores. El caudal y el ángulo del chorro aumentaron con el incremento en la presión.

Aging preferentially affects molecular pathways implicated in development and disease of myelinating glial cells

The central and peripheral nervous systems are involved in multiple agedependent neurological deficits that are often attributed to alterations in function of myelinating glial cells. However, the molecular events that underlie the age-related decline of glial cell function are unknown. We used Schwann cells as a model to study biological processes affected in glial cells by aging. We comprehensively profiled gene expression of the Schwann cell-rich mouse sciatic nerve throughout life, from day of birth until senescence (840 days of age). We combined the aging data with the microarray transcriptional data obtained using nerves isolated from Schwann cell-specific neuropathy-inducing mutants MPZCre/þ/Lpin1fE2-3/fE2-3, MPZCre/þ/ScapfE1/fE1 and Pmp22-null mice. A majority of age related transcripts were also affected in the analyzed mouse models of neuropathy (54.4%) and in development (59.5%) indicating a high level of overlapping in implicated molecular pathways. We observed that compared to peripheral nerve development, dynamically changing expression profiles in aging have opposite (anticorrelated) orientation while they copy the orientation of transcriptional changes observed in analyzed neuropathy models. Subsequent clustering and biological annotation of dynamically changing transcripts revealed that the processes most significantly deregulated in aging include inflammatory/ immune response and lipid biosynthesis/metabolism. Importantly, the changes in these pathways were also observed in myelinated oligodendrocyte- rich optic nerves of aged mice, albeit with lower magnitude. This observation suggests that similar biological processes are affected in aging glial cells in central and peripheral nervous systems, however with different dynamics. Our data, which provide the first comprehensive comparison of molecular changes in glial cells in three distinct biological conditions comprising development, aging and disease, provide not only a new inside into the molecular alterations underlying neural system aging but also identify target pathways for potential therapeutical approaches to prevent or delay complications associated with age-related and inherited forms of neuropathies.

Saccharomyces cerevisiae Grx6 and Grx7 are monothiol glutaredoxins associated with the early secretory pathway

Saccharomyces cerevisiae Grx6 and Grx7 are two monothiol glutaredoxins whose active-site sequences (CSYS and CPYS, respectively) are reminiscent of the CPYC active-site sequence of classical dithiol glutaredoxins. Both proteins contain an N-terminal transmembrane domain which is responsible for their association to membranes of the early secretory pathway vesicles, facing the luminal side. Thus, Grx6 localizes at the endoplasmic reticulum and Golgi compartments, while Grx7 is mostly at the Golgi. Expression of GRX6 is modestly upregulated by several stresses (calcium, sodium, and peroxides) in a manner dependent on the Crz1-calcineurin pathway. Some of these stresses also upregulate GRX7 expression under the control of the Msn2/4 transcription factor. The N glycosylation inhibitor tunicamycin induces the expression of both genes along with protein accumulation. Mutants lacking both glutaredoxins display reduced sensitivity to tunicamycin, although the drug is still able to manifest its inhibitory effect on a reporter glycoprotein. Grx6 and Grx7 have measurable oxidoreductase activity in vivo, which is increased in the presence of tunicamycin. Both glutaredoxins could be responsible for the regulation of the sulfhydryl oxidative state at the oxidant conditions of the early secretory pathway vesicles. However, the differences in location and expression responses against stresses suggest that their functions are not totally overlapping.

A functional variant in the Stearoyl-CoA desaturase gene promoter enhances fatty acid desaturation in pork

There is growing public concern about reducing saturated fat intake. Stearoyl-CoA desaturase (SCD) is the lipogenic enzyme responsible for the biosynthesis of oleic acid (18:1) by desaturating stearic acid (18:0). Here we describe a total of 18 mutations in the promoter and 3′ non-coding region of the pig SCD gene and provide evidence that allele T at AY487830:g.2228T>C in the promoter region enhances fat desaturation (the ratio 18:1/18:0 in muscle increases from 3.78 to 4.43 in opposite homozygotes) without affecting fat content (18:0+18:1, intramuscular fat content, and backfat thickness). No mutations that could affect the functionality of the protein were found in the coding region. First, we proved in a purebred Duroc line that the C-T-A haplotype of the 3 single nucleotide polymorphisms (SNPs) (g.2108C>T; g.2228T>C; g.2281A>G) of the promoter region was additively associated to enhanced 18:1/18:0 both in muscle and subcutaneous fat, but not in liver. We show that this association was consistent over a 10-year period of overlapping generations and, in line with these results, that the C-T-A haplotype displayed greater SCD mRNA expression in muscle. The effect of this haplotype was validated both internally, by comparing opposite homozygote siblings, and externally, by using experimental Duroc-based crossbreds. Second, the g.2281A>G and the g.2108C>T SNPs were excluded as causative mutations using new and previously published data, restricting the causality to g.2228T>C SNP, the last source of genetic variation within the haplotype. This mutation is positioned in the core sequence of several putative transcription factor binding sites, so that there are several plausible mechanisms by which allele T enhances 18:1/18:0 and, consequently, the proportion of monounsaturated to saturated fat.

APOE status modulates the changes in network connectivity induced by brain stimulation in non-demented elders.

Behavioral consequences of a brain insult represent an interaction between the injury and the capacity of the rest of the brain to adapt to it. We provide experimental support for the notion that genetic factors play a critical role in such adaptation. We induced a controlled brain disruption using repetitive transcranial magnetic stimulation (rTMS) and show that APOE status determines its impact on distributed brain networks as assessed by functional MRI (fMRI).Twenty non-demented elders exhibiting mild memory dysfunction underwent two fMRI studies during face-name encoding tasks (before and after rTMS). Baseline task performance was associated with activation of a network of brain regions in prefrontal, parietal, medial temporal and visual associative areas. APOE ε4 bearers exhibited this pattern in two separate independent components, whereas ε4-non carriers presented a single partially overlapping network. Following rTMS all subjects showed slight ameliorations in memory performance, regardless of APOE status. However, after rTMS APOE ε4-carriers showed significant changes in brain network activation, expressing strikingly similar spatial configuration as the one observed in the non-carrier group prior to stimulation. Similarly, activity in areas of the default-mode network (DMN) was found in a single component among the ε4-non bearers, whereas among carriers it appeared disaggregated in three distinct spatiotemporal components that changed to an integrated single component after rTMS. Our findings demonstrate that genetic background play a fundamental role in the brain responses to focal insults, conditioning expression of distinct brain networks to sustain similar cognitive performance.

Diodilaserin käytön edellytykset ohutlevyjen hitsauksessa

Diplomityössä tutkitaan diodilaserhitsausta mahdollisena teollisuuden menetelmänä ja menetelmän vaatimuksia hitsattaessa ohutlevyjä. Työssä tutkittavat materiaalit ovat kylmävalssattu teräs ja ruostumaton teräs sekä liitosmuotoina päittäis-, laippa- ja päällekkäisliitos. Materiaalivahvuudet ovat 0,50 mm:stä 1,50 mm:iin. Työn tavoitteena on määrittää näille kyseisille materiaaleille ja liitosmuodoille hitsausnopeus levynvahvuuden funktiona. Lisäksi käsitellään diodilaserin rakennetta, säteen muodostusta, säteen muokkaamista, säteen analysointia ja säteen turvallisuuteen liittyviä asioita. Suoritetaan vertailua käytössä oleviin muihin lasertyöstömenetelmiin konepajoissa ja tehdään arvio mahdollisen diodilaserinvestoinnin kannattavuudesta. Diodilaserhitsauskokeissa käytettiin Hämeen ammattikorkeakoulun Riihimäen yksikön 1 kW:n tehoista diodilaseria. Koekappaleet leikattiin suuntaisleikkurilla. Osalle hitsatuista kappaleista tehtiin poikittaiset vetokokeet ja mitattiin mikrokovuudet. Virheitä tutkittiin silmämääräisesti sekä radiografisella kuvauksella. Kaikille tutkituille liitoksille, materiaaleille ja vahvuuksille saatiin määriteltyä hitsausnopeudet. Tehtyjen testien perusteella suuntaisleikkurin käyttö on mahdollista. Lisäksi havaittiin suojakaasun käytön myötä, että kirkkaan sulan aiheuttama heijastavuuden kasvu edellyttää hitsausnopeuden pienentämistä.

Gene flow and gene flux shape evolutionary patterns of variation in Drosophila subobscura

Gene flow (defined as allele exchange between populations) and gene flux (defined as allele exchange during meiosis in heterokaryotypic females) are important factors decreasing genetic differentiation between populations and inversions. Many chromosomal inversions are under strong selection and their role in recombination reduction enhances the maintenance of their genetic distinctness. Here we analyze levels and patterns of nucleotide diversity, selection and demographic history, using 37 individuals of Drosophila subobscura from Mount Parnes (Greece) and Barcelona (Spain). Our sampling focused on two frequent O-chromosome arrangements that differ by two overlapping inversions (OST and O3+4), which are differentially adapted to the environment as observed by their opposing latitudinal clines in inversion frequencies. The six analyzed genes (Pif1A, Abi, Sqd, Yrt, Atpa and Fmr1) were selected for their location across the O-chromosome and their implication in thermal adaptation. Despite the extensive gene flux detected outside the inverted region, significant genetic differentiation between both arrangements was found inside it. However, high levels of gene flow were detected for all six genes when comparing the same arrangement among populations. These results suggest that the adaptive value of inversions is maintained, regardless of the lack of genetic differentiation within arrangements from different populations, and thus favors the Local Adaptation hypothesis over the Coadapted Genome hypothesis as the basis of the selection acting on inversions in these populations.

Toiminnanohjauksen kehittäminen toteutusaikataulun avulla

Tässä työssä tarkastellaan projektiin liittyvää toiminnanohjausta. Tutkimusongelmana on, kuinka toiminnanohjausta voitaisiin kehittää aikataulutuksen avulla. Työn yhtenä tavoitteena on luoda kokonaiskuva aikataulutuksen nykytilanteesta. Toisena tavoitteena on kartoittaa ajankohtaiset ongelmat. Kolmantena tavoitteena on luoda malli aikataulun laatimiselle kalanterin valmistuksessa.Kirjallisuustutkimuksen perusteella luotiin kuva projektiliiketoiminnasta ja projektinhallintaan sisältyvästä aikataulunhallinnasta. Haastattelututkimuksen perusteella luotiin kokonaiskuva projektin aikataulutuksesta ja siihen liittyvistä ongelmista. Tutkimusta täydennettiin analysoimalla erään projektin suunniteltua aikataulua ja toteutunutta aikataulua. Toiminnanohjauksen aikataulutustehtävä on muuttumassa yhä tarkeämmäksi toimittaessa resurssien äärirajoilla. Ajankohtaisia kysymyksiä ovat: toteutusaikataulun laatimistekniikka, projektin osittaminen, optimi suunnittelujärjestys ja kriittinen polku.Tutkimuksen perusteella kehitettiin toteutusaikataulun suunnittelun malli ja malliaikataulu. Mallin keskeiset osat ovat: projektin osittaminen tehtäviin, tärkeimpien tehtävien määrittäminen tarkempaa ohjausta varten, tehtävien välisten riippuvuuksien määrittäminen, tehtävien limitysmahdollisuuksien ja ajallisen keston arviointi, aikataulun esittäminen janakaaviona ja ulkoisten tekijöiden, kuten hankinnan asettamien vaatimusten huomioiminen malliaikataulussa. Malliaikataulua tullaan soveltamaan seuraavaan sopivaan toimitusprojektiin.

Puutavarakaupan jakelun nykytila ja kehittäminen pääkaupunkiseudulla

Työn tavoitteena oli kartoittaa kohdeyrityksen pääkaupunkiseudun puu- ja rakennustarvikekaupan jakelun nykytila ja kehittää toimipisteille uusi ja yhtenäinen jakelun toimintamalli. Ensin selvitettiin jakelun nykytila haastattelemalla kuljetuksista vastaavia henkilöitä. Kirjallisuudesta saadun tiedon avulla haettiin vaihtoehtoja uuden jakelumallin luomiselle. Yrityksen tietojärjestelmästä haettiin tietoa toteutuneista kuljetuskustannuksista ja -tapahtumista. Tutkimuksessa selvitettiin myös kuljetuskustannukset ja -tuotot. Tällä hetkellä yritys tekee jakelussa tappiota eli kuljetuskustannukset ovat asiakkailta perittyjä kuljetustuottoja suuremmat. Jokaisessa toimipisteessä kuljetukset hoidetaan itsenäisesti eikä yhteistä järjestelmää ole käytössä. Tästä aiheutuu päällekkäisiä ja turhia ajoja pääkaupunkiseudun alueella, mikä voitaisiin välttää yhteisellä toimintamallilla. Työssä luotiin erilaisia malleja ja niitä verrattiin nykytilanteeseen. Mallien paremmuuden arviointiin käytettiin kannattavuutta ja ulkoista tehokkuutta. Toimintavaihtoehdoiksi nousi kolme eri mallia. Vaihtoehtoina olivat toimipisteiden yhtenäinen malli, jossa ohjaus olisi joko omassa hallinnassa tai ulkoistettu, ja kolmantena vaihtoehtona olisi koko jakelun ulkoistaminen. Malleja varten kartoitettiin eri ohjelmistoja ja ulkoistuspalveluja tarjoavia yrityksiä, jotka olisivat yrityksen tarpeisiin soveltuvia. Toimenpide-ehdotuksina ja suosituksina esitetään yhteiseen jakelujärjestelmään siirtymistä sekä siirtymistä jakelun hinnoittelussa suoriteperusteiseen hinnoitteluun. Tulevaisuudessa jakelun tietoja tulisi kerätä ja tiedon karttuessa laatia suoriteperusteiset hinnoittelutaulukot kuljetuksille. Myös myyjille ja kuljetusvastaaville tarvitaan yhtenäiset toimintaohjeet tilausten muutoksista, kuljetusten hinnoittelusta ja toimituksista.

Yield mapping, soil fertility and tree gaps in an orange orchard

The current high competition on Citrus industry demands from growers new management technologies for superior efficiency and sustainability. In this context, precision agriculture (PA) has developed techniques based on yield mapping and management systems that recognize field spatial variability, which contribute to increase profitability of commercial crops. Because spatial variability is often not perceived the orange orchards are still managed as uniform and adoption of PA technology on citrus farms is low. Thus, the objective of the present study was to characterize the spatial variability of three factors: fruit yield, soil fertility and occurrence of plant gaps caused by either citrus blight or huanglongbing (HLB) in a commercial Valencia orchard in Brotas, São Paulo State, Brazil. Data from volume, geographic coordinates and representative area of the bags used on harvest were recorded to generate yield points that were then interpolated to produce the yield map. Soil chemical characteristics were studied by analyzing samples collected along planting rows and inter-rows in 24 points distributed in the field. A map of density of tree gaps was produced by georeferencing individual gaps and later by counting the number of gaps within 500 m² cells. Data were submitted to statistical and geostatistical analyses. A t test was used to compare means of soil chemical characteristics between sampling regions. High variation on yield and density of tree gaps was observed from the maps. It was also demonstrated overlapping regions of high density of plant absence and low fruit yield. Soil fertility varied depending on the sampling region in the orchard. The spatial variability found on yield, soil fertility and on disease occurrence demonstrated the importance to adopt site specific nutrient management and disease control as tools to guarantee efficiency of fruit production.