Use of constant denaturant capillary electrophoresis of pooled blood samples to identify single-nucleotide polymorphisms in the genes (Scnn1a and Scnn1b) encoding the alpha and beta subunits of the epithelial sodium channel.

BACKGROUND: The epithelial sodium channel (ENaC) is composed of three homologous subunits: alpha, beta, and gamma. Mutations in the Scnn1b and Scnn1g genes, which encode the beta and the gamma subunits of ENaC, cause a severe form of hypertension (Liddle syndrome). The contribution of genetic variants within the Scnn1a gene, which codes for the alpha subunit, has not been investigated. METHODS: We screened for mutations in the COOH termini of the alpha and beta subunits of ENaC. Blood from 184 individuals from 31 families participating in a study on the genetics of hypertension were analyzed. Exons 13 of Scnn1a and Scnn1b, which encode the second transmembrane segment and the COOH termini of alpha- and beta-ENaC, respectively, were amplified from pooled DNA samples of members of each family by PCR. Constant denaturant capillary electrophoresis (CDCE) was used to detect mutations in PCR products of the pooled DNA samples. RESULTS: The detection limit of CDCE for ENaC variants was 1%, indicating that all members of any family or up to 100 individuals can be analyzed in one CDCE run. CDCE profiles of the COOH terminus of alpha-ENaC in pooled family members showed that the 31 families belonged to four groups and identified families with genetic variants. Using this approach, we analyzed 31 rather than 184 samples. Individual CDCE analysis of members from families with different pooled CDCE profiles revealed five genotypes containing 1853G-->T and 1987A-->G polymorphisms. The presence of the mutations was confirmed by DNA sequencing. For the COOH terminus of beta-ENaC, only one family showed a different CDCE profile. Two members of this family (n = 5) were heterozygous at 1781C-->T (T594M). CONCLUSION: CDCE rapidly detects point mutations in these candidate disease genes.

Des "psychoses affectives" aux "caractéristiques psychotiques" : les symptômes psychotiques selon le DSM, discutés à la lumière d'une série de 16 cas de dépression psychotique = From "affective psychoses" to "psychotic features": DSM's psychoticsymptoms discussed in the light of a series of 16 cases of psychoticdepression.

Objectifs. - Le DSM-5 donne une définition du symptôme psychotique indépendante de tout concept depsychose. Chez les patients dépressifs, la présence d'hallucination ou d'idée délirante, quelle que soit leurforme clinique, conduit en principe à diagnostiquer une dépression psychotique et à prescrire des neurolep-tiques. Les symptômes psychotiques sont subdivisés par le DSM en « congruents » ou « non congruents » àl'humeur. Nous discutons de la pertinence d'une catégorie de symptômes psychotiques « atypiques », peuévocateurs d'une psychose au sens classique du terme. Méthode. - Discussion de la définition opérationnelle des symptômes psychotiques du DSM, étude d'unesérie de 16 patients chez qui un diagnostic de dépression psychotique a été posé. Résultats. - Sur les 16 patients, deux seulement présentaient des symptômes psychotiques classiques, évo-cateurs d'une psychose. Chez les autres, le diagnostic reposait sur la présence de symptômes très atypiques,comme des hallucinations visuelles par exemple.

Light-mediated polarization of the PIN3 auxin transporter for the phototropic response in Arabidopsis.

Phototropism is an adaptation response, through which plants grow towards the light. It involves light perception and asymmetric distribution of the plant hormone auxin. Here we identify a crucial part of the mechanism for phototropism, revealing how light perception initiates auxin redistribution that leads to directional growth. We show that light polarizes the cellular localization of the auxin efflux carrier PIN3 in hypocotyl endodermis cells, resulting in changes in auxin distribution and differential growth. In the dark, high expression and activity of the PINOID (PID) kinase correlates with apolar targeting of PIN3 to all cell sides. Following illumination, light represses PINOID transcription and PIN3 is polarized specifically to the inner cell sides by GNOM ARF GTPase GEF (guanine nucleotide exchange factor)-dependent trafficking. Thus, differential trafficking at the shaded and illuminated hypocotyl side aligns PIN3 polarity with the light direction, and presumably redirects auxin flow towards the shaded side, where auxin promotes growth, causing hypocotyls to bend towards the light. Our results imply that PID phosphorylation-dependent recruitment of PIN proteins into distinct trafficking pathways is a mechanism to polarize auxin fluxes in response to different environmental and endogenous cues.

Test-retest repeatability of the pupil light response to blue and red light stimuli in normal human eyes using a novel pupillometer.

In this study, we evaluated the repeatability of pupil responses to colored light stimuli in healthy subjects using a prototype chromatic pupillometer. One eye of 10 healthy subjects was tested twice in the same day using monochromatic light exposure at two selected wavelengths (660 and 470 nm, intensity 300 cd/m(2)) presented continuously for 20 s. Pupil responses were recorded in real-time before, during, and after light exposure. Maximal contraction amplitude and sustained contraction amplitude were calculated. In addition, we quantified the summed pupil response during continuous light stimulation as the total area between a reference line representing baseline pupil size and the line representing actual pupil size over 20 s (area under the curve). There was no significant difference in the repeated measure compared to the first test for any of the pupil response parameters. In conclusion, we have developed a novel prototype of color pupillometer which demonstrates good repeatability in evoking and recording the pupillary response to a bright blue and red light stimulus.

Light framing, estructura lleugera; un camí per la estandarització en el procés constructiu. Cas general del sistema light framing i en cas particular del sistema eurodom

Projecte d'una vivenda unifamiliar amb un sistema constructiu mitjançant estructura lleugera on el material principal és la fusta i altres materials sostenibles i mètodes d'execució de prefabricació o in situ.

Drought tolerance and light requirements of high and low sublittoral species of Mediterranean macroalgae of the genus Cystoseira C. Agardh (Fucales, Phaeophyceae)

Article sobre la tolerància a la sequera i les necessitats de llum d'espècies de l'alt i baix sublitoral de macroalgues de la Mediterrània del gènere Cystoseira C. Agardh (Fucales, Phaeophyceae)

Kuluttajapakkauskartonkien laserleikattavuuden karakterisointi

Tämän diplomityön tarkoituksena oli karakterisoida kuluttajapakkauskartonkien laserleikattavuutta. Kirjallisuusosassa perehdyttiin asiaa käsittelevään kirjallisuuteen ja tutkimuksiin ja kokeellisessa osassa käsiteltiin kuitumateriaalien laserleikkausta erilaisin leikkausparametrein. Leikattujen näytteiden leikkausrailosta otetuista mikroskooppikuvista määritettiin leikkausrailon uraleveys, purseen ja mustumisen määrä. Työssä tutkitut materiaalit olivat koivu- ja mäntysellu, CTMP, päällystämätön ja päällystetty sellukartonki sekä päällystämätön CTMP-runkoinen nestepakkauskartonki. Työssä havaittiin, että tutkitut kuitumateriaalit soveltuivat erinomaisesti laserleikkaukseen. Leikattujen kuitumateriaalinäytteiden leikkausrailo oli hyvälaatuinen eli leikkausrailo oli kapea, täydellisesti näytteen lävitse, tasainen reunoiltaan, väriltään vaalea ( ei hiiltynyt ) ja ei sisältänyt railon reunassa pystyssä olevia katkenneita kuituja ( pursetta ). Työssä todettiin myös, että kuitumateriaaleja laserleikatessa leikkausnopeus ja laserteho riippuivat lineaarisesti toisistaan. Leikkausnopeuden kasvaessa tarvittiin enemmän lasertehoa hyvälaatuisen leikkausrailon saavuttamiseksi. Tällöin leikkausuran leveys pysyi lähes vakiona. Lisäksi huomattiin, että optimisijainti polttopisteelle olisi 0.1-0.4 mm materiaalin pinnan yläpuolella, vaikka kirjallisuudesta löydettiin suosituksia sijoittaa polttopiste materiaalin pinnan alapuolelle. Näillä polttopisteen sijainnin arvoilla saavutettiin suurimmat leikkausnopeudet ja kapeimmat urat.

Pectina extraída de casca de pequi e aplicação em geleia light de manga

Dentre os frutos do Cerrado, destaca-se o pequi (Caryocar brasiliense Camb.), que é constituído por aproximadamente 80% de casca, que é desprezada; no entanto, apresenta potencial de utilização em várias aplicações. O objetivo deste trabalho foi avaliar a influência das variáveis concentração de ácido cítrico, temperatura e tempo de extração sobre o rendimento e o grau de esterificação da pectina extraída da casca de pequi e compará-la com a pectina cítrica comercial aplicada na formulação de geleia light. Obtiveram-se rendimentos de pectina entre 14,89 e 55,86 g.100g-1. A pectina obtida da casca de pequi caracterizou-se por apresentar baixo grau de esterificação (11,79-48,87%). A geleia light elaborada a partir da pectina da casca de pequi, extraída à temperatura de 84ºC por 92 minutos, na presença de 2% de ácido cítrico, obteve boa aceitação por parte dos provadores, alcançando escores médios acima de 7,0, diferindo da geleia produzida com pectina cítrica comercial apenas na aparência. Conclui-se que é viável utilizar a pectina da casca de pequi como ingrediente para formulação de geleia light de manga.

Label-free detection of tobramycin in serum by transmission-localized surface plasmon resonance.

In order to improve the efficacy and safety of treatments, drug dosage needs to be adjusted to the actual needs of each patient in a truly personalized medicine approach. Key for widespread dosage adjustment is the availability of point-of-care devices able to measure plasma drug concentration in a simple, automated, and cost-effective fashion. In the present work, we introduce and test a portable, palm-sized transmission-localized surface plasmon resonance (T-LSPR) setup, comprised of off-the-shelf components and coupled with DNA-based aptamers specific to the antibiotic tobramycin (467 Da). The core of the T-LSPR setup are aptamer-functionalized gold nanoislands (NIs) deposited on a glass slide covered with fluorine-doped tin oxide (FTO), which acts as a biosensor. The gold NIs exhibit localized plasmon resonance in the visible range matching the sensitivity of the complementary metal oxide semiconductor (CMOS) image sensor employed as a light detector. The combination of gold NIs on the FTO substrate, causing NIs size and pattern irregularity, might reduce the overall sensitivity but confers extremely high stability in high-ionic solutions, allowing it to withstand numerous regeneration cycles without sensing losses. With this rather simple T-LSPR setup, we show real-time label-free detection of tobramycin in buffer, measuring concentrations down to 0.5 μM. We determined an affinity constant of the aptamer-tobramycin pair consistent with the value obtained using a commercial propagating-wave based SPR. Moreover, our label-free system can detect tobramycin in filtered undiluted blood serum, measuring concentrations down to 10 μM with a theoretical detection limit of 3.4 μM. While the association signal of tobramycin onto the aptamer is masked by the serum injection, the quantification of the captured tobramycin is possible during the dissociation phase and leads to a linear calibration curve for the concentrations over the tested range (10-80 μM). The plasmon shift following surface binding is calculated in terms of both plasmon peak location and hue, with the latter allowing faster data elaboration and real-time display of the results. The presented T-LSPR system shows for the first time label-free direct detection and quantification of a small molecule in the complex matrix of filtered undiluted blood serum. Its uncomplicated construction and compact size, together with the remarkable performances, represent a leap forward toward effective point-of-care devices for therapeutic drug concentration monitoring.