Comparative transcriptomics of extreme phenotypes of human HIV-1 infection and SIV infection in sooty mangabey and rhesus macaque.

High levels of HIV-1 replication during the chronic phase of infection usually correlate with rapid progression to severe immunodeficiency. However, a minority of highly viremic individuals remains asymptomatic and maintains high CD4+ T cell counts. This tolerant profile is poorly understood and reminiscent of the widely studied nonprogressive disease model of SIV infection in natural hosts. Here, we identify transcriptome differences between rapid progressors (RPs) and viremic nonprogressors (VNPs) and highlight several genes relevant for the understanding of HIV-1-induced immunosuppression. RPs were characterized by a specific transcriptome profile of CD4+ and CD8+ T cells similar to that observed in pathogenic SIV-infected rhesus macaques. In contrast, VNPs exhibited lower expression of interferon-stimulated genes and shared a common gene regulation profile with nonpathogenic SIV-infected sooty mangabeys. A short list of genes associated with VNP, including CASP1, CD38, LAG3, TNFSF13B, SOCS1, and EEF1D, showed significant correlation with time to disease progression when evaluated in an independent set of CD4+ T cell expression data. This work characterizes 2 minimally studied clinical patterns of progression to AIDS, whose analysis may inform our understanding of HIV pathogenesis.

Tumor targeting with newly designed biparatopic antibodies directed against two different epitopes of the carcinoembryonic antigen (CEA).

In an attempt to improve tumor targeting and tumor retention time of monoclonal antibodies (MAbs), we prepared biparatopic antibodies (BpAbs) having the capability of binding 2 different non-overlapping epitopes on the same target antigen molecule, namely, the carcinoembryonic antigen (CEA). Six BpAbs were constructed by coupling 2 different Fab' fragments from 4 different specific anti-CEA MAbs recognizing 4 CEA epitopes (Gold 1-4). Demonstration of the double paratopic binding of these antibodies for CEA was confirmed in vitro by inhibition radioimmunoassay and cross-inhibition analysis by surface plasmon resonance (SPR; BIACORE) technology. Using the latter technique, the affinity constants for CEA immobilized onto the sensor chip were found to range from 0.37 to 1.54 x 10(9) M(-1) for the 4 parental F(ab')2 fragments and from 1.88 to 10.14 x 10(9) M(-1) for the BpAbs, demonstrating the advantage of biparatopic binding over conventional F(ab')2 binding. The Ka improvement was particularly high for BpAb F6/35A7 and BpAb F6/B17 with a 9.5- and 8.1-fold increase, respectively, as compared with the parental F(ab')2. In vivo, the 6 BpAbs were compared with their 2 respective parental F(ab')2 by injection of 131I-BpAb/125I-F(ab')2 parental fragments into nude mice xenografted with the human colon carcinoma T380. Dissection 72 hr post-injection demonstrated that BpAb B17/CE25 and BpAb F6/B17 gave higher tumor uptake than that of their parental F(ab')2. This finding is particularly interesting for BpAb F6/B17, which compared favorably with the F6 F(ab')2, one of the best parental F(ab')2 fragments used in our study.

Korkeavaaleuksisen PGW-massan valmistaminen

Työssä haettiin optimaalista valkaisukemikaalien annostelusuhdetta mahdollisimman vaalean massan valmistamiseksi. Tarkoituksena oli myös selvittää käytettävän tehdasprosessin pullonkauloja ja esittää vaihtoehtoja prosessin kehittämiseksi. Kirjallisuusosassa tutustuttiin mekaanisen massan peroksidivalkaisuun ja erilaisiin prosessimalleihin sen suorittamiseksi. Kirjallisuudesta saatuja lähtötietoja sovellettiin sitten ensin laboratorio- ja laajemmin tehdaskokein käytäntöön. Laboratoriokokeissa haettiin optimaalista lipeäannosta vakioperoksidiannoksella. Lisäksi selvitettiin lämpötilan ja viipymäajan vaikutusta valkaisutulokseen. Tuloksista oli todettavissa, että sekä lämpötilan että lipeäannoksen kasvattaminen kiihdyttää valkaisureaktiota. Korkeammassa lämpötilassa tarvittava lipeäannos on pienempi. Lyhyellä viipymäajalla ja matalammalla lämpötilalla saavutetaan hyviä tuloksia vain suurella lipeäannoksella. Suurempaa silikaattiannosta käytettäessä valkaisun jälkeen mitattu loppupH oli korkeampi ja jäännösperoksidin määrä hieman suurempi kuin referenssipisteessä. Vaaleudessa ei merkittävää muutosta näkynyt. Yleisesti laboratoriokokeiden tulokset vastasivat kirjallisuudessa esitettyjä tuloksia. Tehdaskokeet suoritettiin kahdella peroksidiannoksella. Kummallekin peroksidiannokselle haettiin optimaalinen lipeäannos. Lisäksi seurattiin silikaattiannoksen vaikutusta syntyvään vaaleuteen. Varsinaisten kokeiden lisäksi suoritettiin lyhyt maksimivaaleuskoe suurella peroksidiannoksella käyttäen kahta eri lipeäannosta. Suurin mitattu vaaleus oli 79,3 % ISO. Peroksidiannoksella 2,5 % saavutettiin noin 14 ISO-yksikön vaaleudennousu, ja annoksella 3 % vaaleudennousu oli noin 16 yksikköä. Tehdaskokeiden aikana kokeiltiin myös kemikaalien laimennusveden määrän vaikutusta vaaleuteen. Veden määrän vähetessä valkaisusakeus nousi ja vaaleus parani. Hiomon nykyprosessille laadittiin aine- ja energiatase. Taseet tehtiin myös prosessille, jossa nykyvalkaisimoon on lisätty valkaistun massan pesu ja jäännöskemikaalien kierrätys. Taseiden tarkoituksena oli selvittää virtaavien jakeiden määriä ja prosessin energiatasapainoa eri tilanteissa. Massan puhtauden paranemista pesun aikana tarkasteltiin laskennallisesti. Tämän työn aikana kävi selväksi, että oikeilla valkaisukemikaalisuhteilla pystytään valmistamaan vaaleaa massaa. Viipymäaikaa lisäämällä valkaisuun saisi lisää tehoa, mutta vain tunnin viipymäajalla päästään jo hyviin tuloksiin. Vaaleusheittoja aiheuttavat valkaistavan massan lähtövaaleuden, sakeuden ja lämpötilan muutokset. Lisäksi viipymäajan muutokset aiheuttavat huojuntaa saavutettavaan loppuvaaleuteen.

Johdon ja keskijohdon rekrytointiprosessin kehittäminen Suomessa Case: UPM-Kymmene Oyj

Tutkielman tarkoituksena on olla kehittämässä UPM-Kymmenen johdon ja keskijohdon rekrytointiprosessia. Tavoitteena on tutkia osaamisniukkuuden aiheuttaman riskin minimoimista, hyvien työntekijöiden palkkaamista ja kuinka työntekijät saadaan pysymään yhtiön palveluksessa. Tutkimusmenetelmä on kvalitatiivinen. Se perustuu muun muassa rekrytointiryhmän kokousten materiaaleihin ja muihin sisäisiin raportteihin sekä haastatteluun. Johdon ja keskijohdon kohdalla mahdollisia rekrytoitavia on vähän, joten heidän pysyvyytensä yrityksen palveluksessa tulee varmistaa. Uuden työntekijän löytäminen on usein vaikeaa ja rekrytointikustannukset ovat suuria. Osaamisniukkuutta voidaan välttää hyvällä henkilöstösuunnittelulla.

Stabilization of the ionic form of weak acid cation exchange resins

Weak acid cation exchange (WAC) resins are used in the chromatographic separation of betaine from vinasse, a by-product of sugar industry. The ionic form of the resin determines the elution time of betaine. When a WAC-resin is in hydrogen form, the retention time of betaine is the longest and betaine elutes as the last component of vi-nasse from the chromatographic column. If the feed solution contains salts and its pH is not acidic enough to keep the resin undissociated, the ionic form of the hydrogen form resin starts to alter. Vinasse contains salts and its pH is around 5, it also contains weak acids. To keep the metal ion content (Na/H ratio) of the resin low enough to ensure successful separation of betaine, acid has to be added to either eluent (water) or vinasse. The aim of the present work was to examine by laboratory experiments which option requires less acid. Also the retention mechanism of betaine was investigated by measuring retention volumes of acetic acid and choline in different Na/H ratios of the resin. It was found that the resulting ionic form of the resin is the same regardless of whether the regeneration acid is added to the eluent or the feed solution (vinasse). Be-sides the salt concentration and the pH of vinasse, also the concentration of weak acids in the feed affects the resulting ionic form of the resin. The more buffering capacity vinasse has, the more acid is required to keep the ionic form of the resin desired. Vinasse was found to be quite strong buffer solution, which means relatively high amounts of acid are required to prevent the Na/H ratio from increasing too much. It is known that the retention volume of betaine decreases significantly, when the Na/H ratio increases. This is assumed to occur, because the amount of hydrogen bonds between the carboxylic groups of betaine and the resin decreases. Same behavior was not found with acetic acid. Choline has the same molecular structure as betaine, but instead of carboxylic group it has hydroxide group. The retention volume of choline increased as the Na/H ratio of the resin increased, because of the ion exchange reaction between choline cation and dissociated carboxylic group of the resin. Since the retention behavior of choline on the resin is opposite to the behavior of be-taine, the strong affinity of betaine towards hydrogen form WAC-resin has to be based on its carboxylic group. It is probable that the quaternary ammonium groups also affect the behavior of the carboxylic groups of betaine, causing them to form hydrogen bonds with the carboxylic groups of the resin.

Determinação de 2,5-hexanodiona em urina empregando cromatografia líquida de alta eficiência, após derivatização com 2,4-dinitrofenil-hidrazina

A method for quantifying urinary 2,5-hexanedione was optimized and validated. Urine samples were hydrolyzed and derivatized with 2,4-dinitrophenylhydrazine. The analyte was separated in a high performance liquid chromatography system with a diode array detector, using a C18 column (150 x 4.6 mm, p.d. 5 µm) and a mobile phase composed of phosphate buffer pH 2.3:acetonitrile (40:60, v/v), at a flow rate of 1 mL/min. The chromatograms were monitored at 334 nm. Retention time was 7.3 minutes. Main validation parameters were: coefficient of determination: 0.9994, accuracy: 96 to 107%; intra-assay precision (RSD): 3.08 to 6.72%; inter-assay precision (RSD): 2.54 to 8.17% and limit of quantitation of 0.19 µg/mL.

Desenvolvimento de metodologia para avaliar remoção de estrogênios em estações de tratamento de esgotos

The present research aimed to study the parameters that affect the determination of estrogens in sewage upon the use of SPE and determination by using gas chromatography mass spectrometry (GC/MS). The study evaluated the stabilization pond technology with regard to estrogens removal. The results showed that the methodology was able to identify and quantify the estrogens E1, E2, E2-17A and EE2 in sewage. Moreover, the estrogens sometimes were not removed in the stabilization pond, showing that even the high hydraulic retention time (HRT) and high sunlight intensity were not enough to degrade these compounds completely during load peaks.

Kaasun dispergoituminen tehdasmittakaavaisessa keskisakean massan sekoittimessa

Työn tarkoituksena oli selvittää Proto 10 kemikaalisekoittimen kyky dispergoida kaasua keskisakean valkaistun mäntysellun joukkoon. Työssä käytettiin kuvantamistekniikkaa, jonka avulla pystyttiin näkemään sekoittimen ja putkiston sisälle sekoitustapahtuman aikana. Muodostunutta dispersiota tarkasteltiin kolmesta kuvauspisteestä, joista yksi sijaitsi sekoittimen pesässä ja kaksi putkistossa sekoittimen jälkeen. Työssä verrattiin myös laboratoriosekoittimella saatuja tuloksia teollisen mittakaavan sekoittimella saatuihin tuloksiin, sekä määritettiin tarvittava pinta-aktiivisen aineen konsentraatio, jolla saavutettiin ruskeaa massaa vastaava vaahtoaminen. Työn kokeellinen osuus koostui kolmesta osasta. Ensimmäisessä vaiheessa tutkittiin ruskeasta mänty- ja koivumassasta lingotun suodoksen vaahtoamista ja verrattiin sitä vedellä ja pesuaineella saatavaan vaahtoon. Toisessa vaiheessa suoritettiin referenssiajot laboratoriosekoittimen ja teollisen mittakaavan sekoittimen vertailua varten Quantum Mark IV laboratoriosekoittimella. Kolmannessa vaiheessa tutkittiin Proto 10 sekoittimen kykyä dispergoida kaasua mäntysellun joukkoon eri pyörimisnopeuksilla ja virtaamilla. Työn tuloksien perusteella energiankäytön kannalta paras pyörimisnopeus Proto 10 sekoittimelle on 1500 min-1. Nostamalla kierrosnopeutta yli tämän ei saavutettu merkittävää parannusta dispersion laadussa varsinkaan suuremmilla virtaamilla. Virtaamalla todettiin olevan suuri merkitys sekoitustulokseen. Laboratoriosekoittimella tehtyjen kokeiden todettiin vastaavan parhaiten teollisen mittakaavan tuloksia pyörimisnopeudella 1200 min-1.

Performance of UASB reactors in two stages under different HRT and OLR treating residual waters of swine farming

In this study it was evaluated the effects of hydraulic retention time (HRT) and Organic Loading Rate (OLR) on the performance of UASB (Upflow Anaerobic Sludge Blanket) reactors in two stages treating residual waters of swine farming. The system consisted of two UASB reactors in pilot scale, installed in series, with volumes of 908 and 188 L, for the first and second stages (R1 and R2), respectively. The HRT applied in the system of anaerobic treatment in two stages (R1 + R2) was of 19.3, 29.0 and 57.9 h. The OLR applied in the R1 ranged from 5.5 to 40.1 kg CODtotal (m³ d)-1. The average removal efficiencies of chemical oxygen demand (COD) and total suspended solids (TSS) ranged, respectively, from 66.3 to 88.2% and 62.5 to 89.3% in the R1, and from 85.5 to 95.5% and 76.4 to 96.1% in the system (R1 + R2). The volumetric production of methane in the system (R1 + R2) ranged from 0.295 to 0.721 m³CH4 (m³ reactor d)-1. It was found that the OLR applied were not limiting to obtain high efficiencies of CODtotal and TSS removal and methane production. The inclusion of the UASB reactor in the second stage contributed to increase the efficiencies of CODtotal and TSS removal, especially, when the treatment system was submitted to the lowest HRT and the highest OLR.

Swine effluent treatment using anaerobic digestion at different loading rates

The industrial swine production is characterized by generation of significant effluent amounts that require treatment. The most adopted practices by Brazilian swine farmers have been wastewater storage in lagoons and its subsequent use as a biofertilizer. Nutrient accumulation in soil and water creates the need for an effective management of these residues. The anaerobic digestion process is an important alternative and low-cost treatment for organic matter reduction. However, its efficiency is limited by the digester capacity of solid degradation, especially at low hydraulic retention times. Thus, the present study aimed to verify the behavior of an upflow anaerobic digester by increasing the organic loading rate. This was accomplished in three stages using, as a parameter, volatile solids at 0.5; 1.0 and 1.5 kgVS m-3 d-1, respectively. This digester model proved to be quite robust and effective in swine manure treatment, achieving high efficiency of volatile solid removal at all stages of the study (stage 1: 61.38%; stage 2: 55.18%; and stage 3: 43.18%). Biogas production was directly related to the increasing organic load, reaching 0.14, 0.85, and 0.86 Nm³ kgVS-1add., respectively, with no significant difference (p<0.05) of biogas methane concentration among the studied stages (73.7, 75.0, and 77.9%).

Potential of biogas production from swine manure supplemented with glycerine waste

In this study, was studied the biogas generation from swine manure, using residual glycerine supplementation. The biogas production by digestion occurred in the anaerobic batch system under mesophilic conditions (35°C), with a hydraulic retention time of 48 days. The experiment was performed with 48 samples divided into four groups, from these, one was kept as a control (without glycerin) and the other three groups were respectively supplemented with residual glycerine in the percentage of 3%, 6% and 9% of the total volume of the samples. The volume of biogas was controlled by an automated system for reading in laboratory scale and the quality of the biogas (CH4) measured from a specific sensor. The results showed that the residual glycerine has high potential for biogas production, with increases of 124.95%, 156.98% and 197.83% in the groups 3%, 6% and 9%, respectively, relative to the sample control. However, very high organic loads can compromise the process of digestion affecting the quality of the biogas generated in relation to methane.

Start-up of anaerobic reactors for slaughterhouse wastewater treatment

This study aimed to evaluate the start-up of a horizontal anaerobic fixed bed reactor (HAFBR) followed by an upflow anaerobic sludge blanket (UASB) for the slaughterhouse wastewater treatment. HAFBR was filled with bamboo rings and had 1.2 m in length, 0.10 m in diameter and volume of 7.5 L. The UASB had the volume of 15 L. The HAFBR and UASB operated at organic loading rate and hydraulic retention time average of 8.46 and 3.77 kg m-3 d-1 of COD and 0.53 and 0.98 days, respectively. During 150 days of monitoring system it was found pH 6.8, relatively high values of bicarbonate alkalinity (> 1000 mg L-1) and reduced values of volatile acids (70 to 150 mg L-1), which afforded average removal efficiencies of COD total and total suspended solids of the order of 31 and 23% in HAFBR and 79% and 63% in UASB. It can be concluded that the generation and consumption of bicarbonate alkalinity and total volatile acids, thereby maintaining the pH during the study indicated stable operation of the reactors. The COD removal in the reactors was satisfactory especially when it considers that the assessment was conducted in a period of adaptation of organisms to the effluent and also the high organic load applied during this period.

Hydrogen production from cassava processing wastewater in an anaerobic fixed bed reactor with bamboo as a support material

Attempting to associate waste treatment to the production of clean and renewable energy, this research sought to evaluate the biological production of hydrogen using wastewater from the cassava starch treatment industry, generated during the processes of extraction and purification of starch. This experiment was carried out in a continuous anaerobic reactor with a working volume of 3L, with bamboo stems as the support medium. The system was operated at a temperature of 36°C, an initial pH of 6.0 and under variations of organic load. The highest rate of hydrogen production, of 1.1 L.d-1.L-1, was obtained with application of an organic loading rate of 35 g.L-1.d-1, in terms of total sugar content and hydraulic retention time of 3h, with a prevalence of butyric and acetic acids as final products of the fermentation process. Low C/N ratios contributed to the excessive growth of the biomass, causing a reduction of up to 35% in hydrogen production, low percentages of H2 and high concentrations of CO2in the biogas.

Stability of Natural Colorants of Plant Origin

In recent years, there have been studies that show a correlation between the hyperactivity of children and use of artificial food additives, including colorants. This has, in part, led to preference of natural products over products with artificial additives. Consumers have also become more aware of health issues. Natural food colorants have many bioactive functions, mainly vitamin A activity of carotenoids and antioxidativity, and therefore they could be more easily accepted by the consumers. However, natural colorant compounds are usually unstable, which restricts their usage. Microencapsulation could be one way to enhance the stability of natural colorant compounds and thus enable better usage for them as food colorants. Microencapsulation is a term used for processes in which the active material is totally enveloped in a coating or capsule, and thus it is separated and protected from the surrounding environment. In addition to protection by the capsule, microencapsulation can also be used to modify solubility and other properties of the encapsulated material, for example, to incorporate fat-soluble compounds into aqueous matrices. The aim of this thesis work was to study the stability of two natural pigments, lutein (carotenoid) and betanin (betalain), and to determine possible ways to enhance their stability with different microencapsulation techniques. Another aim was the extraction of pigments without the use of organic solvents and the development of previously used extraction methods. Stability of pigments in microencapsulated pigment preparations and model foods containing these were studied by measuring the pigment content after storage in different conditions. Preliminary studies on the bioavailability of microencapsulated pigments and sensory evaluation for consumer acceptance of model foods containing microencapsulated pigments were also carried out. Enzyme-assisted oil extraction was used to extract lutein from marigold (Tagetes erecta) flower without organic solvents, and the yield was comparable to solvent extraction of lutein from the same flowers. The effects of temperature, extraction time, and beet:water ratio on extraction efficiency of betanin from red beet (Beta vulgaris) were studied and the optimal conditions for maximum yield and maximum betanin concentration were determined. In both cases, extraction at 40 °C was better than extraction at 80 °C and the extraction for five minutes was as efficient as 15 or 30 minutes. For maximum betanin yield, the beet:water ratio of 1:2 was better, with possibly repeated extraction, but for maximum betanin concentration, a ratio of 1:1 was better. Lutein was incorporated into oil-in-water (o/w) emulsions with a polar oil fraction from oat (Avena sativa) as an emulsifier and mixtures of guar gum and xanthan gum or locust bean gum and xanthan gum as stabilizers to retard creaming. The stability of lutein in these emulsions was quite good, with 77 to 91 percent of lutein being left after storage in the dark at 20 to 22°C for 10 weeks whereas in spray dried emulsions the retention of lutein was 67 to 75 percent. The retention of lutein in oil was also good at 85 percent. Betanin was incorporated into the inner w1 water phase of a water1-in-oil-inwater2 (w1/o/w2) double emulsion with primary w1/o emulsion droplet size of 0.34 μm and secondary w1/o/w2 emulsion droplet size of 5.5 μm and encapsulation efficiency of betanin of 89 percent. In vitro intestinal lipid digestion was performed on the double emulsion, and during the first two hours, coalescence of the inner water phase droplets was observed, and the sizes of the double emulsion droplets increased quickly because of aggregation. This period also corresponded to gradual release of betanin, with a final release of 35 percent. The double emulsion structure was retained throughout the three-hour experiment. Betanin was also spray dried and incorporated into model juices with different pH and dry matter content. Model juices were stored in the dark at -20, 4, 20–24 or 60 °C (accelerated test) for several months. Betanin degraded quite rapidly in all of the samples and higher temperature and a lower pH accelerated degradation. Stability of betanin was much better in the spray dried powder, with practically no degradation during six months of storage in the dark at 20 to 24 °C and good stability also for six months in the dark at 60 °C with 60 percent retention. Consumer acceptance of model juices colored with spray dried betanin was compared with similar model juices colored with anthocyanins or beet extract. Consumers preferred beet extract and anthocyanin colored model juices over juices colored with spray dried betanin. However, spray dried betanin did not impart any off-odors or off-flavors into the model juices contrary to the beet extract. In conclusion, this thesis describes novel solvent-free extraction and encapsulation processes for lutein and betanin from plant sources. Lutein showed good stability in oil and in o/w emulsions, but slightly inferior in spray dried emulsions. In vitro intestinal lipid digestion showed a good stability of w1/o/w2 double emulsion and quite high retention of betanin during digestion. Consumer acceptance of model juices colored with spray dried betanin was not as good as model juices colored with anthocyanins, but addition of betanin to real berry juice could produce better results with mixture of added betanin and natural berry anthocyanins could produce a more acceptable color. Overall, further studies are needed to obtain natural colorants with good stability for the use in food products.

Typenpoiston tehostaminen kunnallisessa jätevedenpuhdistuksessa membraanibioreaktorin avulla

Tämä diplomityö tehtiin Vihdin Vesihuoltolaitoksen Nummelan jäteveden puhdistamolle. Työssä tutkittiin typenpoistoa kunnallisista jätevesistä membraanibioreaktorin (MBR) avulla. MBR:ssä yhdistyvät perinteinen aktiivilieteprosessi ja kalvosuodatus. Työn tavoite oli päästä yli 95 % typenpoistoon. Aluksi typenpoisto oli yli 80 %, kun pilot-mittakaavan MBR-laitosta operoitiin perinteisen prosessin parametrein. Typenpoistoa onnistuttiin tehostamaan nostamalla nitraattipitoisen palautuslietteen kierrätystä prosessin alkupäähän (1600 L/h) ja lisäämällä aktiivista biomassaa reaktorissa. Yli 90 % typenpoisto edellytti myös pidempää viipymäaikaa (noin kaksinkertainen perinteiseen prosessiin verrattuna). Tutkimuksessa päästiin parhaimmillaan jopa 95 % typenpoistumaan operoimalla laitteistoa pienellä typpikuormalla (0,1 kg/vrk) ja alhaisemmalla lietepitoisuudella (10 g/L). Typpikuorman noustessa (0,3 kg/vrk) typenpoistoteho laski. Tätä onnistuttiin parantamaan (yli 90 %) nostamalla biomassan määrää reaktorissa (15 g/L). Hyvän typenpoiston saavuttaminen edellytti myös suurempaa metanolin ja hapen syöttöä.