Regulation of nonphosphorylated and phosphorylated forms of neurofilament proteins in the prefrontal cortex of human opioid addicts.

The neurofilament (NF) proteins (NF-H, NF-M, and NF-L for high, medium, and low molecular weights) play a crucial role in the organization of neuronal shape and function. In a preliminary study, the abundance of total NF-L was shown to be decreased in brains of opioid addicts. Because of the potential relevance of NF abnormalities in opioid addiction, we quantitated nonphosphorylated and phosphorylated NF in postmortem brains from 12 well-defined opioid abusers who had died of an opiate overdose (heroin or methadone). Levels of NF were assessed by immunoblotting techniques using phospho-independent and phospho-dependent antibodies, and the relative (% changes in immunoreactivity) and absolute (changes in ng NF/microg total protein) amounts of NF were calculated. Decreased levels of nonphosphorylated NF-H (42-32%), NF-M (14-9%) and NF-L (30-29%) were found in the prefrontal cortex of opioid addicts compared with sex, age, and postmortem delay-matched controls. In contrast, increased levels of phosphorylated NF-H (58-41%) and NF-M (56-28%) were found in the same brains of opioid addicts. The ratio of phosphorylated to nonphosphorylated NF-H in opioid addicts (3.4) was greater than that in control subjects (1.6). In the same brains of opioid addicts, the levels of protein phosphatase of the type 2A were found unchanged, which indicated that the hyperphosphorylation of NF-H is not the result of a reduced dephosphorylation process. The immunodensities of GFAP (the specific glial cytoskeletol protein), alpha-internexin (a neuronal filament related to NF-L) and synaptophysin (a synapse-specific protein) were found unchanged, suggesting a lack of gross changes in glial reaction, other intermediate filaments of the neuronal cytoskeletol, and synaptic density in the prefrontal cortex of opioid addicts. These marked reductions in total NF proteins and the aberrant hyperphosphorylation of NF-H in brains of opioid addicts may play a significant role in the cellular mechanisms of opioid addiction.

Velocity-jump instabilities in Hele-Shaw flow of associating polymer solutions

We study fracturelike flow instabilities that arise when water is injected into a Hele-Shaw cell filled with aqueous solutions of associating polymers. We explore various polymer architectures, molecular weights, and solution concentrations. Simultaneous measurements of the finger tip velocity and of the pressure at the injection point allow us to describe the dynamics of the finger in terms of the finger mobility, which relates the velocity to the pressure gradient. The flow discontinuities, characterized by jumps in the finger tip velocity, which are observed in experiments with some of the polymer solutions, can be modeled by using a nonmonotonic dependence between a characteristic shear stress and the shear rate at the tip of the finger. A simple model, which is based on a viscosity function containing both a Newtonian and a non-Newtonian component, and which predicts nonmonotonic regions when the non-Newtonian component of the viscosity dominates, is shown to agree with the experimental data.

Polydispersity index from linear viscoelastic data: Unimodal and bimodal linear polymer melts

This article describes a method for determining the polydispersity index Ip2=Mz/Mw of the molecular weight distribution (MWD) of linear polymeric materials from linear viscoelastic data. The method uses the Mellin transform of the relaxation modulus of a simple molecular rheological model. One of the main features of this technique is that it enables interesting MWD information to be obtained directly from dynamic shear experiments. It is not necessary to achieve the relaxation spectrum, so the ill-posed problem is avoided. Furthermore, a determinate shape of the continuous MWD does not have to be assumed in order to obtain the polydispersity index. The technique has been developed to deal with entangled linear polymers, whatever the form of the MWD is. The rheological information required to obtain the polydispersity index is the storage G′(ω) and loss G″(ω) moduli, extending from the terminal zone to the plateau region. The method provides a good agreement between the proposed theoretical approach and the experimental polydispersity indices of several linear polymers for a wide range of average molecular weights and polydispersity indices. It is also applicable to binary blends.

Postnatal expression pattern of calcium-binding proteins in organotypic thalamic cultures and in the dorsal thalamus in vivo.

The present study describes the postnatal expression of calbindin, calretinin and parvalbumin and glutamic acid decarboxylase (GAD) and microtubule-associated protein 2 (MAP2) in organotypic monocultures of rat dorsal thalamus compared to the thalamus in vivo. Cultures were maintained for up to 7 weeks. Cortex-conditioned medium improved the survival of thalamic cultures. MAP2-immunoreactive material was present in somata and dendrites of small and large-sized neurons throughout the cultures. Parvalbumin immunoreactivity was present in larger multipolar or bitufted neurons along the edge of a culture. These neurons also displayed strong parvalbumin mRNA and GAD mRNA expression, and GABA immunoreactivity. They likely corresponded to cells of the nucleus reticularis thalami. Parvalbumin mRNA, but neither parvalbumin protein nor GAD mRNA, was expressed in neurons with large somata within the explant. They likely represented relay cells. GAD mRNA, but not parvalbumin mRNA, was expressed in small neurons within the explants. Small neurons also displayed calbindin- and calretinin-immunoreactivity. The small neurons likely represented local circuit neurons. The time course of expression of the calcium-binding proteins revealed that all were present at birth with the predicted molecular weights. A low, but constant parvalbumin expression was observed in vitro without the developmental increase seen in vivo, which most likely represented parvalbumin from afferent sources. In contrast, the explantation transiently downregulated the calretinin and calbindin expression, but the neurons recovered the expression after 14 and 21 days, respectively. In conclusion, thalamic monocultures older than three weeks represent a stable neuronal network containing well differentiated neurons of the nucleus reticularis thalami, relay cells and local circuit neurons.

MAP2 Isoforms in Developing Cat Cerebral Cortex and Corpus Callosum.

The microtubule-associated protein MAP2 was studied in the developing cat visual cortex and corpus callosum. Biochemically, no MAP2a was detectable in either structure during the first postnatal month; adult cortex revealed small amounts of MAP2a. MAP2b was abundant in cortical tissue during the first postnatal month and decreased in concentration towards adulthood; it was barely detectable in corpus callosum at all ages. MAP2c was present in cortex and corpus callosum at birth; in cortex it consisted of three proteins of similar molecular weights between 65 and 70 kD. The two larger, phosphorylated forms disappeared after postnatal day 28, the smaller form after day 39. In corpus callosum, MAP2c changed from a phosphorylated to an unphosphorylated variant during the first postnatal month and then disappeared. Immunocytochemical experiments revealed MAP2 in cell bodies and dendrites of neurons in all cortical layers, from birth onwards. In corpus callosum, in the first month after birth, a little MAP2, possibly MAP2c, was detectable in axons. The present data indicate that MAP2 isoforms differ in their cellular distribution, temporal appearance and structural association, and that their composition undergoes profound changes during the period of axonal stabilization and dendritic maturation.

Identification of promising antigenic components in latent fingermark residues

An analysis of latent fingermark residues by Sodium-Dodecyl-Sulfate PolyAcrylamide Gel Electrophoresis (SDS-PAGE) followed by silver staining allowed the detection of different proteins, from which two major bands, corresponding to proteins of 56 and 64 kDa molecular weight, could be identified. Two other bands, corresponding to proteins of 52 and 48 kDa were also visualizable along with some other weaker bands of lower molecular weights. In order to identify these proteins, three antibodies directed against human proteins were tested on western blots of fingermarks residues: anti-keratin 1 and 10 (K1/10), anti-cathepsin-D (Cat.D) and anti-dermcidin (Derm.). The corresponding antigens are known to be present in the stratum corneum of desquamating stratified epithelium (K1/10, Cat.D) and/or in eccrine sweat (Cat.D, Derm.). The two major bands were identified as consistent with keratin 1 and 10. The pro-form and the active form of the cathepsin-D have also been identified from two other bands. Dermcidin could not be detected in the western blot. In addition, these antibodies have been tested on latent fingermarks left on polyvinylidene fluoride (PVDF) membrane, as well as on whitened and non-whitened paper. The detection of fingermarks was successful with all three antibodies.

Unsaturated Polyester Resins from Recycled Polyethyleneterephthalate

Tämän diplomityön tarkoituksena oli tutkia miten kuluttajien kierrättämästä polyeteenitereftalaatista ( PET ) voi valmistaa tyydyttymättömiä polyesterihartseja. Työssä valmistettiin yleiskäyttöön soveltuva laminointihartsi sekä 'gel coat' -hartsi jota käytetään esim. veneiden pintamaalina. Yleishartsin depolymerointiin käytettiin propyleeniglykolia ja 'gel coat' -hartsin valmistamiseen neopentyyliglykolia. Polykondensaatiovaiheessa reaktioon lisättiin maleiinihappoa ja lopuksi hartsit liuotettiin styreeniin. Kirjallisuusosassa esitetään eri menetelmiä PET:n depolymeroimiseksi. Lisäksi esitetään eri vaihtoehtoja glykolien, happojen, katalyyttien ja vinyylimonomeerien valitsemiseksi tyydyttymättömien polyesterihartsien valmistuksessa. Analyysimenetelmiä nestemäisten ja kovetettujen hartsien tutkimiseen ja vertailuun käydään läpi kuten myös erilaisia sovelluksia polyesterihartsien käyttämiseksi. Kokeellinen osa todisti että PET-pullojäte voidaan prosessoida hartsiksiilman uusia investointeja prosessilaitteistoon. PET:n glykolyysi kesti viidestäseitsemään tuntia ja polykondensaatiovaihe kahdesta ja puolesta viiteen tuntiin. Hartsien molekyylipainot ja mekaanisten testien tulokset olivat vertailukelpoisia kaupallisten hartsien antamien tulosten kanssa. Glykolyysivaiheen momomeeri- ja oligomeeripitoisuudet mitattiin geelipermeaatiokromatografialla, jotta nähtiin miten pitkälle depolymerisaatio oli edennyt. Tätä tietoa voidaan hyödyntää uusien hartsireseptin suunnittelussa. Polymeeriketjussa jäljellä olevien C=C kaksoissidosten määrä ja niiden isomeraatioaste maleaattimuodosta fumaraattimuotoon mitattiin 1H-NMR -menetelmällä. Tislevesien koostumus määritettiin kaasukromatografialla, ja tulokset kertoivat katalyytin sisältämän kloorin reagoineen glykolien kanssa, johtaen suureen glykolikulutukseen ja muihin ei-toivottuihin sivureaktioihin. Hartsien sietokykyä auringon valolle mitattiin niiden UV-absorption avulla. Kummastakin hartsista valmistettiin 'gel coat' -maalit jotkalaitettiin sääkoneeseen, joka simuloi auringonpaistetta ja vesisadetta vuorotellen. Näistä 'gel coateista' mitattiin niiden kellastumista. Kummastakin hartsista tehdyt valut asetettiin myös sääkoneeseen ja IR-spektreistä ennen jajälkeen koetta nähtiin että C=O ja C-O esterisidoksia oli hajonnut.

Proteins influencing foam formation in wine and beer: the role of yeast

This review focuses on the role of proteins in the production and maintenance of foam in both sparkling wines and beer. The quality of the foam in beer but especially in sparkling wines depends, among other factors, on the presence of mannoproteins released from the yeast cell walls during autolysis. These proteins are hydrophobic, highly glycosylated, and their molecular masses range from 10 to 200 kDa characteristics that allow mannoproteins to surround and thus stabilize the gas bubbles of the foam. Both the production and stabilization of foam also depend on other proteins. In wine, these include grape-derived proteins such as vacuolar invertase; in beer, barley-derived proteins, such as LTP1, protein Z, and hordein-derived polypeptides, are even more important in this respect than mannoproteins

Study on the removal of biodegradable NOM from seawater using biofiltration

Despite the low biodegradability of seawater NOM, problems associated with biofouling are common in facilities that handle seawater. In this work, a fixed-film aerobic biofilter is proposed as an effective unit for preventing biofouling in such facilities. A packed-bed biofilter with an EBCT = 6 - 11 min was employed. The results demonstrated that the DOC is reduced by 6% and the BOD7 is reduced up to 15%. The LC-OCD analysis revealed that biofiltration abates the LMW neutrals and biopolymer fractions by 33 and 17%, respectively. However, the fractionation with UF membrane showed that the biofiltration process is able to degrade the more biodegradable compounds that have molecular weights that are greater than 1 kDa and compounds with molecular weights of less than 1 kDa. After biofiltration, the biological activity measured in terms of ATP removal was reduced by 60%. Finally, a test to evaluate the biofilm formation capacity of a water sample revealed reductions of ~94% when comparing biofiltered and non-biofiltered seawater. Therefore, a fixed-film aerobic biofiltration process could be a useful treatment for the removal of biodegradable organic matter from seawater and for improving the water quality in terms of less biofilm formation capacity.

Ultrafiltration membranes from waste polyethylene terephthalate and additives: synthesis and characterization

The synthesis and characterization of asymmetric ultrafiltration membranes from recycled polyethylene terephthalate (PET) and polyvinylpyrrolidone (PVP) is reported. PET is currently used in many applications, including the manufacture of bottles and tableware. Monomer extraction from waste PET is expensive, and this process has not yet been successfully demonstrated on a viable scale. Hence, any method to recycle or regenerate PET once it has been used is of significant importance from scientific and environmental research viewpoints. Such a process would be a green alternative due to reduced raw monomer consumption and the additional benefit of reduced manufacturing costs. The membranes described here were prepared by a phase-inversion process, which involved casting a solution containing PET, m-cresol as solvent, and polyethylene glycol (PEG) of different molecular weights as additives. The membranes were characterized in terms of pure water permeability (PWP), molecular weight cut-off (MWCO), and flux and membrane morphology. The results show that the addition of PEG with high molecular weights leads to membranes with higher PWP. The presence of additives affects surface roughness and membrane morphology.

Öljynjalostamon vetyverkon PINCH-analyysi

Tässä diplomityössä on tarkasteltu Porvoon öljynjalostamon vetyverkkoa ja pohdittu keinoja, joilla vedyn käyttöä jalostamolla voitaisiin tehostaan sekä polttokaasuverkkoon menevän vedyn määrä pienentää. Tarkastelun lähtökohtana toimii vetytaseen pohjalta laadittu vetypinch-analyysi. Kirjallisuusosassa on esitelty jalostamon vetyverkkoon kuuluvat yksiköt sekä käsitelty lyhyesti niiden toimintaa. Lisäksi on käsitelty vetypinch-analyysin periaate, sekä kuinka todelliset prosessirajoitteet voidaan huomioida sitä toteutettaessa. Kirjallisuusosan lopussa on esitetty miten vetyverkon vaiheittainen optimointi etenee. Työn soveltavassa osassa laadittiin vetyverkon virtauskaavio, jolla saatiin luotua kattava käsitys jalostamon vedynjakelusta. Virtauskaaviosta tehtiin yksinkertaistettu versio, jonka perusteella laadittiin vetytase. Vetytaseen pohjalta suoritettiin vetypinch-analyysi, jonka mukaan jalostamolla tuotettiin tasehetkellä ylimäärin vetyä. Vedyn käytön tehostamiseksi jalostamolla tulee rikkivedyn talteenottoyksikkö 2:n polttokaasuvirta pyrkiä minimoimaan tai hyödyntämään. Lisäksi virtausmittareiden mitoituspisteiden molekyylimassat tulisi muuttaa vastaamaan paremmin nykyistä ajotilannetta, sekä seurata niitä jatkossa säännöllisesti. Myös vetypitoisuutta mittaavien online-analysaattoreiden kalibroinnista tulee huolehtia, ja ottaa riittävästi kenttänäytteitä vetyverkosta. On huomattava, että öljynjalostamon vedyn tuotannon minimointi ei ole aina automaattisesti taloudellisin ratkaisu. Joissain tapauksissa vedyn osapaineen nostaminen vetyä kuluttavan yksikön reaktorissa voi lisätä yksikön tuottavuutta niin paljon, että se kompensoi lisääntyneestä vedyn tuotannosta aiheutuvat kustannukset.

The husk fiber of Cocos nucifera L. (Palmae) is a source of anti-neoplastic activity

In the present study, we investigated the in vitro anti-tumoral activities of fractions from aqueous extracts of the husk fiber of the typical A and common varieties of Cocos nucifera (Palmae). Cytotoxicity against leukemia cells was determined by the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay. Cells (2 x 104/well) were incubated with 0, 5, 50 or 500 µg/mL high- or low-molecular weight fractions for 48 h, treated with MTT and absorbance was measured with an ELISA reader. The results showed that both varieties have almost similar antitumoral activity against the leukemia cell line K562 (60.1 ± 8.5 and 47.5 ± 11.9% for the typical A and common varieties, respectively). Separation of the crude extracts with Amicon membranes yielded fractions with molecular weights ranging in size from 1-3 kDa (fraction A) to 3-10 kDa (fraction B) and to more than 10 kDa (fraction C). Cells were treated with 500 µg/mL of these fractions and cytotoxicity was evaluated by MTT. Fractions ranging in molecular weight from 1-10 kDa had higher cytotoxicity. Interestingly, C. nucifera extracts were also active against Lucena 1, a multidrug-resistant leukemia cell line. Their cytotoxicity against this cell line was about 50% (51.9 ± 3.2 and 56.3 ± 2.9 for varieties typical A and common, respectively). Since the common C. nucifera variety is extensively cultured in Brazil and the husk fiber is its industrial by-product, the results obtained in the present study suggest that it might be a very inexpensive source of new antineoplastic and anti-multidrug resistant drugs that warrants further investigation.

Potential of quixaba (Sideroxylon obtusifolium) latex as a milk-clotting agent

There are several obstacles to the use of chymosin in cheese production. Consequently, plant proteases have been studied as possible rennet substitutes, but most of these enzymes are unsuitable for the manufacture of cheese. The aim of this study was to evaluate the potential of latex from Sideroxylon obtusifolium as a source of milk-clotting proteases and to partially characterize the enzyme. The enzyme extract showed high protease and coagulant activities, with an optimal pH of 8.0 and temperature of 55 °C. The enzyme was stable in wide ranges of temperature and pH. Its activity was not affected by any metal ions tested; but was inhibited by phenylmethanesulfonyl fluoride and pepstatin. For the coagulant activity, the optimal concentration of CaCl2 was 10 µmol L- 1. Polyacrylamide gel electrophoresis showed four bands, with molecular weights between 17 and 64 kDa. These results indicate that the enzyme can be applied to the cheese industry.

Peptide separation of commercial fermented milk during refrigerated storage

Milk is an important source of bioactive compounds. Many of these compounds are released during fermentation and refrigerated storage. The aim of this study was to determine the release of peptides by lactic acid bacteria in commercial fermented milk during refrigerated storage. The size and profile of peptides were analyzed by polyacrylamide gel electrophoresis and sizeexclusion HPLC. During electrophoresis, it was observed that the peptides were released from caseins, whereas β-lactoglobulin was the whey protein with the highest degradation. HPLC analysis confirmed the pattern of peptide formation observed in electrophoresis. Two fractions lower than 2 kDa with aromatic amino acids in their structure were separated. These results were consistent with those reported for structures of peptides with antihypertensive activity. Therefore, the presence of aromatic amino acids in the peptide fractions obtained increases the likelihood of finding peptides with such activity in refrigerated commercial fermented milk. In conclusion, during cold storage, peptides with different molecular weights are released and accumulated. This could be due to the action of proteinases and peptidases of the proteolytic system in lactic acid bacteria.

Investigation of the composition of linear alkylbenzenes with emphasis on the identification and quantitation of some trace compounds using GS/MS system in both electron impact and chemical ionization modes

Linear alkylbenzenes, LAB, formed by the Alel3 or HF catalyzed alkylation of benzene are common raw materials for surfactant manufacture. Normally they are sulphonated using S03 or oleum to give the corresponding linear alkylbenzene sulphonates In >95 % yield. As concern has grown about the environmental impact of surfactants,' questions have been raised about the trace levels of unreacted raw materials, linear alkylbenzenes and minor impurities present in them. With the advent of modem analytical instruments and techniques, namely GCIMS, the opportunity has arisen to identify the exact nature of these impurities and to determine the actual levels of them present in the commercial linear ,alkylbenzenes. The object of the proposed study was to separate, identify and quantify major and minor components (1-10%) in commercial linear alkylbenzenes. The focus of this study was on the structure elucidation and determination of impurities and on the qualitative determination of them in all analyzed linear alkylbenzene samples. A gas chromatography/mass spectrometry, (GCIMS) study was performed o~ five samples from the same manufacturer (different production dates) and then it was followed by the analyses of ten commercial linear alkylbenzenes from four different suppliers. All the major components, namely linear alkylbenzene isomers, followed the same elution pattern with the 2-phenyl isomer eluting last. The individual isomers were identified by interpretation of their electron impact and chemical ionization mass spectra. The percent isomer distribution was found to be different from sample to sample. Average molecular weights were calculated using two methods, GC and GCIMS, and compared with the results reported on the Certificate of Analyses (C.O.A.) provided by the manufacturers of commercial linear alkylbenzenes. The GC results in most cases agreed with the reported values, whereas GC/MS results were significantly lower, between 0.41 and 3.29 amu. The minor components, impurities such as branched alkylbenzenes and dialkyltetralins eluted according to their molecular weights. Their fragmentation patterns were studied using electron impact ionization mode and their molecular weight ions confirmed by a 'soft ionization technique', chemical ionization. The level of impurities present i~ the analyzed commercial linear alkylbenzenes was expressed as the percent of the total sample weight, as well as, in mg/g. The percent of impurities was observed to vary between 4.5 % and 16.8 % with the highest being in sample "I". Quantitation (mg/g) of impurities such as branched alkylbenzenes and dialkyltetralins was done using cis/trans-l,4,6,7-tetramethyltetralin as an internal standard. Samples were analyzed using .GC/MS system operating under full scan and single ion monitoring data acquisition modes. The latter data acquisition mode, which offers higher sensitivity, was used to analyze all samples under investigation for presence of linear dialkyltetralins. Dialkyltetralins were reported quantitatively, whereas branched alkylbenzenes were reported semi-qualitatively. The GC/MS method that was developed during the course of this study allowed identification of some other trace impurities present in commercial LABs. Compounds such as non-linear dialkyltetralins, dialkylindanes, diphenylalkanes and alkylnaphthalenes were identified but their detailed structure elucidation and the quantitation was beyond the scope of this study. However, further investigation of these compounds will be the subject of a future study.