Analysis of the contribution of the beta-oxidation auxiliary enzymes in the degradation of the dietary conjugated linoleic acid 9-cis-11-trans-octadecadienoic acid in the peroxisomes of Saccharomyces cerevisiae.

Beta-oxidation of the conjugated linoleic acid 9-cis,11-trans-octadecadienoic acid (rumenic acid) was analyzed in vivo in Saccharomyces cerevisiae by monitoring polyhydroxyalkanoate production in the peroxisome. Polyhydroxyalkanoate is synthesized by the polymerization of the beta-oxidation intermediates 3-hydroxyacyl-CoAs via a bacterial polyhydroxyalkanoate synthase targeted to the peroxisome. The amount of polyhydroxyalkanaote synthesized from the degradation of rumenic acid was found to be similar to the amount synthesized from the degradation of 10-trans,12-cis-octadecadienoic acid, oleic acid or 10-cis-heptadecenoic acid. Furthermore, the degradation of 10-cis-heptadecenoic acid was found to be unaffected by the presence of rumenic acid in the media. Efficient degradation of rumenic acid was found to be independent of the Delta(3,5),Delta(2,4)-dienoyl-CoA isomerase but instead relied on the presence of Delta(3),Delta(2)-enoyl-CoA isomerase activity. The presence of the unsaturated monomer 3-hydroxydodecenoic acid in polyhydroxyalkanoate derived from rumenic acid degradation was found to be dependent on the presence of a Delta(3),Delta(2)-enoyl-CoA isomerase activity. Together, these data indicate that rumenic acid is mainly degraded in vivo in S. cerevisiae through a pathway requiring only the participation of the auxiliary enzymes Delta(3),Delta(2)-enoyl-CoA isomerase, along with the enzyme of the core beta-oxidation cycle.

Molecular interactions involved in the transactivation of the human T-cell leukemia virus type 1 promoter mediated by Tax and CREB-2 (ATF-4).

The human T-cell leukemia virus type 1 (HTLV-1) Tax protein activates viral transcription through three 21-bp repeats located in the U3 region of the HTLV-1 long terminal repeat and called Tax-responsive elements (TxREs). Each TxRE contains nucleotide sequences corresponding to imperfect cyclic AMP response elements (CRE). In this study, we demonstrate that the bZIP transcriptional factor CREB-2 is able to bind in vitro to the TxREs and that CREB-2 binding to each of the 21-bp motifs is enhanced by Tax. We also demonstrate that Tax can weakly interact with CREB-2 bound to a cellular palindromic CRE motif such as that found in the somatostatin promoter. Mutagenesis of Tax and CREB-2 demonstrates that both N- and C-terminal domains of Tax and the C-terminal region of CREB-2 are required for direct interaction between the two proteins. In addition, the Tax mutant M47, defective for HTLV-1 activation, is unable to form in vitro a ternary complex with CREB-2 and TxRE. In agreement with recent results suggesting that Tax can recruit the coactivator CREB-binding protein (CBP) on the HTLV-1 promoter, we provide evidence that Tax, CREB-2, and CBP are capable of cooperating to stimulate viral transcription. Taken together, our data highlight the major role played by CREB-2 in Tax-mediated transactivation.

Estudos Cinéticos da aquação do trans-[Co(en)2Cl2]Cl

The trans-dichlorobis(ethylenediamine)cobalt(III) chloride was synthesized in an undergraduate laboratory and its aquation reaction was carried out at different temperatures. This reaction follows pseudo-first-order kinetics and the rate constants, determined at 25, 35, 45, 55 and 70 º C, are 1.44 x 10-3; 5.14 x 10-3; 1.48 x 10-2; 4.21 x 10-2 and 2.21 x 10-1 s-1, respectively. The activation energy is 93.99 ± 2.88 kJ mol-1.

Optimization of a HPLC procedure for simultaneous determination of cisplatin and the complex cis,cis,trans-diamminedichlorodihydroxoplatinum(IV) in aqueous solutions

A RP-HPLC procedure for the simultaneous determination of cisplatin and the complex cis,cis,trans-diamminedichlorodihydroxo-platinum(IV), was development. The developed procedure was validated in terms of linearity, accuracy, precision, limits of detection (LOD), limits of quantification (LOQ) and specificity. The limits of detection (LOD) were 0.47 x 10-4 and 0.53 x 10-4 mol L-1 and the limits of quantification (LOQ) were 1.57 x 10-4 and 1.75 x 10-4 mol L-1, for cisplatin and cis,cis,trans-diamminedichlorodihydroxopla-tinum(IV), respectively. The average recoveries of cisplatin and cis,cis,trans-diamminedichlorodihydroxoplatinum(IV) was 100.6% ± 1.4 and 101.2% ± 1.1, respectively. Intermediate (inter-day) precision, repeatability and specificity of the procedure for hydrolysis products of cisplatin were studied. The results of the study showed that the proposed RP-HPLC procedure is simple, rapid, precise, accurate and specific.

Exploración estocástica de las superficies de energía potencial de dímeros cis-trans y trans-trans del ácido fórmico

Potential energy surface (PES) of cis-trans and trans-trans formic acid dimers were sampled using a stochastic method, and the geometries, energies, and vibrational frequencies were computed at B3LYP/6-311++G(3df,2p) level of theory. The results show that molar free energy of dimerization deviated up to 108.4% when basis set superposition error (BSSE) and zero-point energy (ZPE) were not considered. For cis-trans dimers, C=O and O - H bond weakened, whereas C - O bonds strengthened due to dimerization. Also, trans-trans FA dimers did not show a trend regarding strengthening or weakening of the C=O, O - H and C - O bonds.

Esofagectomia trans-hiatal no tratamento do megaesôfago chagásico avançado

OBJETIVO: Avaliar os resultados da esofagectomia trans-hiatal no tratamento do megaesôfago chagásico avançado. MÉTODO: Foram estudados retrospectivamente 28 pacientes portadores de megaesôfago chagásico avançado (MCA), graus III e IV, segundo a classificação radiológica de Rezende (adotada pela Organização Mundial de Saúde), e que foram submetidos à esofagectomia subtotal trans-hiatal no Serviço de Clínica Cirúrgica do Hospital Universitário Prof. Alberto Antunes (HUPAA) da Universidade Federal de Alagoas, entre 1982 e 2000. Foram analisadas, as seguintes variáveis: A) Queixas clínicas pré-operatórias versus as pós-operatórias (disfagia, regurgitação, pirose, diarréia, dumping, plenitude pós-prandial, pneumonia e o estado ponderal). B) avaliação radiológica pós-operatória da boca anastomótica esofagogástrica cervical e do estômago transposto. C) avaliação endoscópica pós-operatória do coto esofágico e da boca anastomótica. RESULTADOS: O seguimento variou de 4 a 192 meses, média de 58,18 meses. Dezesseis pacientes eram do sexo feminino e 12 masculinos. Idade mínima de 16 e máxima de 67 anos, média de 36,5 anos. Não houve mortalidade nesta série. Houve resolução plena da disfagia na maioria dos pacientes (20/28 - 71,4%), um (3,6%) referiu disfagia leve que não necessitou tratamento e 7/28 (25%) necessitaram de uma ou mais sessões de dilatação. Nenhum necessitou de dilatação permanente. A pirose foi o sintoma mais importante no seguimento tardio (35,7%), seguida da regurgitação (25%), diarréia (14,3%), plenitude pós-prandial (10,7%) e dumping (3,6%). Houve ganho ponderal em 87,5% dos pacientes avaliados. A esofagite no coto esofágico foi o achado endoscópico mais significativo (46,4%). O esôfago de Barrett no coto remanescente foi encontrada em 10,7% dos casos. A maioria dos achados radiológicos foi normal, embora três doentes (10,7%) tenham apresentado estase gástrica. CONCLUSÃO: A esofagectomia trans-hiatal mostrou-se eficaz para o tratamento da disfagia no megaesôfago chagásico avançado, embora com morbidade elevada, porém com mortalidade nula.

Esofagectomia trans-hiatal versus transtorácica: experiência do Instituto Nacional do Câncer (INCA)

OBJETIVO: Analisar comparativamente a morbimortalidade e sobrevida após esofagectomia trans-hiatal (TH) ou transtorácica (TT). METODOS: Estudo retrospectivo não randomizado de 68 pacientes com neoplasia de esôfago operados no INCA entre 1997 e 2005, divididos em dois grupos: 1 - TH (33 pacientes); e 2 - TT (35 pacientes). RESULTADOS: A idade média foi 40,7 anos (25 - 74 anos), sendo 73,5% homens. Tumores do 1/3 médio predominaram no Grupo 2 (48,6% versus 21,2%, p = 0,02). A média de linfonodos dissecados foi maior no Grupo 2 (21,6 versus 17,8 linfonodos, p = 0,04), porém sem diferença no número de linfonodos metastáticos (4,1 versus 3,9 linfonodos, p = 0,85). O tempo cirúrgico médio foi maior no Grupo 2 (410 versus 270 minutos, p = 0,001). O tempo médio de internação também foi maior no Grupo 2 (19 versus 14 dias, p = 0,001). A morbidade operatória foi 50%, sem diferença significativa (42,4% versus 57,1%, p = 0,23). Fístula esofágica ocorreu em 13,2%, sem diferença significativa (9,1% versus 17,1%, p = 0,23). A mortalidade foi 5,8% (04 pacientes), sem diferença significativa (1,4% versus 4,4%, p = 0,83). CONCLUSÃO: Neste estudo, a morbimortalidade não apresentou diferença em relação à via de acesso para a esofagectomia, apesar do maior tempo cirúrgico e de permanência hospitalar na via TT. A sobrevida global em 3 e 5 anos também foi maior na TT, possivelmente devido a maior freqüência de estágios iniciais em pacientes submetidos à transtorácica.

Valtatien 4 parantaminen Äänekosken kohdalla : Ympäristövaikutusten arviointiselostus

Tämä YVA-selostus koskee valtatie 4:n parantamista Äänekosken kohdalla. Suunnittelualue käsittää Huutomäen (valtatien 13 risteys) ja Mämmen kylän välisen alueen. YVA-menettelyn tavoitteena on, että menettelyn avulla valtatielle 4 Äänekosken kohdalla löydetään mahdollisimman hyvä kokonaisratkaisu. YVA-menettelyn yhteydessä on myös laadittu alustava yleissuunnitelma ja hankearviointi. Valtatie 4 on osa kansainvälistä Eurooppa-teiden verkkoa (Trans-European-Network). Valtatie 4:lle Äänekosken kohdalla on asetettu tavoitetila valtatie 4 Jyväskylä–Oulu-yhteysvälin kehittämisselvityksessä. Selvityksen mukaan valtatie 4 on tavoitetilassa kapea nelikaistainen keskikaiteellinen tie välillä valtatie 13 (Huutomäki)–Äänekoski. Keski-Suomen maakuntakaavassa valtatie 4 on esitetty Vehniän ja Äänekosken välillä ohjeellisena moottoritienä. Valtatien 4 parantaminen on lähtenyt liikkeelle Äänekosken kaupungin tarpeesta laatia kaupungin kehittämistarpeita tukeva tilavaraus valtatielle. Lisäksi tavoitteena on parantaa liikenteen sujuvuutta ja liikenneturvallisuutta suunnittelualueella. YVA-selostusvaiheessa on arvioitu kolmea eri toteutusvaihtoehtoa, vaihtoehdot 0, 1 ja 2. Vaihtoehto 0 on nykyinen valtatie 4:n pääosin yksiajoratainen linjaus nykyisen tieverkon mukaisesti. Vaihtoehtoon ei ole suunniteltu toimenpiteitä. Vaihtoehto 1 on kaksiajoratainen, moottoritietasoinen tie, jonka nopeusrajoitus on 100 km/h ja liittymät ovat eritasoliittymiä. Vaihtoehto 2 on pääosin nykyisen tien maastokäytävää noudattava kaksiajoratainen tie, jonka nopeusrajoitus on 80 km/h–100 km/h ja jonka liittymät ovat eritasoliittymiä. Vaihtoehdoille 1 ja 2 on laadittu alustavat yleissuunnitelmat. Ympäristövaikutusten arviointi on tehty YVA-lain ja laaditun ympäristövaikutusten arviointiohjelman mukaisesti huomioiden yhteysviranomaisen YVA-ohjelmasta antamassaan lausunnossaan esittämät täydennykset ja täydennyksiä varten laaditut vastineet. Selostusvaiheessa on arvioitu hankkeen vaikutukset aluerakenteeseen ja maankäyttöön; luonnonoloihin ja suojelualueisiin; pinta- ja pohjavesiin; maa- ja kallioperään sekä luonnonvarojen käyttöön; maisemaan ja kulttuuriympäristöön; melu- ja päästövaikutukset sekä ihmisten elinoloihin ja viihtyvyyteen. Vaikutuksia arvioitaessa huomioitiin sekä tien käytön että rakentamisen aikaiset vaikutukset. Lisäksi arvioitiin hankkeen liikenteelliset vaikutukset. Jatkosuunnitteluun valittavasta vaihtoehdosta voidaan tehdä päätös yhteysviranomaisen lausunnon antamisen jälkeen. Valitusta vaihtoehdosta laaditaan maantielain mukaiset yleissuunnitelma ja tiesuunnitelma. Tiesuunnitteluvaiheen arvioidaan alkavan aikaisintaan vuonna 2016. Rakentaminen edellyttää hyväksyttyä tiesuunnitelmaa ja alkaa siten aikaisintaan vuoden 2020 jälkeen. Rakentaminen kestää alustavien arvioiden mukaan 2-3 vuotta.

Valtatien 4 parantaminen Äänekosken kohdalla : Hankearviointi

Tämä hankearviointi koskee valtatie 4:n parantamista Äänekosken kohdalla. Suunnittelualue käsittää Huutomäen (valtatien 13 liittymä) ja Mämmen kylän välisen alueen. Hankearviointi on tehty kiinteässä yhteistyössä YVA-menettelyn ja sen yhteydessä tehdyn alustavan yleissuunnitelman laatimisen kanssa. Hankearviointi tukee päätöksentekoa suunnitelmavaihtoehtojen arviointivaiheessa. Valtatie 4 on osa kansainvälistä Eurooppa-teiden verkkoa (Trans-European-Network). Valtatie 4:lle Äänekosken kohdalla on asetettu tavoitetila valtatie 4 Jyväskylä–Oulu-yhteysvälin kehittämisselvityksessä. Selvityksen mukaan valtatie 4 on tavoitetilassa kapea nelikaistainen keskikaiteellinen tie välillä valtatie 13 (Huutomäki)–Äänekoski. Keski-Suomen maakuntakaavassa valtatie 4 on esitetty Vehniän ja Äänekosken välillä ohjeellisena moottoritienä. Valtatien 4 parantaminen on lähtenyt liikkeelle Äänekosken kaupungin tarpeesta laatia kaupungin kehittämistarpeita tukeva tilavaraus valtatielle. Lisäksi tavoitteena on parantaa liikenteen sujuvuutta ja liikenneturvallisuutta suunnittelualueella. Hankearvioinnissa on arvioitu kolmea eri toteutusvaihtoehtoa, vaihtoehdot 0, 1 ja 2. Vaihtoehto 0 on nykyinen valtatie 4:n pääosin yksiajoratainen linjaus nykyisen tieverkon mukaisesti. Vaihtoehtoon ei ole suunniteltu toimenpiteitä. Vaihtoehto 1 on kaksiajoratainen, moottoritietasoinen tie, jonka nopeusrajoitus on 100 km/h ja liittymät ovat eritasoliittymiä. Vaihtoehto 2 on pääosin nykyisen tien maastokäytävää noudattava kaksiajoratainen tie, jonka nopeusrajoitus on 80 km/h–100 km/h ja jonka liittymät ovat eritasoliittymiä. Vaihtoehto 1 on kannattavuudeltaan paras (H/K=1,8). Vaihtoehto 2 ei ole kannattava (H/K=0,8). Jatkosuunnitteluun valittavasta vaihtoehdosta voidaan tehdä päätös yhteysviranomaisen lausunnon antamisen jälkeen. Valitusta vaihtoehdosta laaditaan maantielain mukaiset yleissuunnitelma ja tiesuunnitelma. Tiesuunnitteluvaiheen arvioidaan alkavan aikaisintaan vuonna 2016. Rakentaminen edellyttää hyväksyttyä tiesuunnitelmaa ja alkaa siten aikaisintaan vuoden 2020 jälkeen. Rakentaminen kestää alustavien arvioiden mukaan 2-3 vuotta.

Cardiovascular responses to microinjection of trans-(±)-ACPD into the NTS were similar in conscious and chloralose-anesthetized rats

The changes in mean arterial pressure (MAP) and heart rate (HR) in response to the activation of metabotropic receptors in the nucleus tractus solitarii (NTS) with trans-(±)-1-amino-1,3-cyclopentanedicarboxylic acid (trans-(±)-ACPD) were evaluated in conscious and anesthetized Wistar, male rats weighing 240-260 g (N = 8). The responses obtained with trans-(±)-ACPD were compared with the responses to L-glutamate (1 nmol/100 nl), since in a previous study we showed that anesthesia converted a pressor response to L-glutamate microinjected into the NTS of conscious rats to a depressor response in the same rats under urethane or chloralose anesthesia. Microinjection of 3 doses of trans-(±)-ACPD (100, 500 and 1000 pmol/100 nl) produced a dose-dependent fall in MAP (range, -20 to -50 mmHg) and HR (range, -30 to -170 bpm) under both conscious and chloralose anesthesia conditions. These data indicate that the cardiovascular responses to the activation of metabotropic receptors by trans-(±)-ACPD are not affected by chloralose anesthesia while the cardiovascular responses to the activation of excitatory amino acid (EAA) receptors by L-glutamate are significantly altered

Effect of all-trans retinoic acid on newly diagnosed acute promyelocytic leukemia patients: results of a Brazilian center

Thirty-seven patients with acute promyelocytic leukemia (APL) were treated with all-trans retinoic acid (ATRA). Patients received 45 mg m-2 day-1 po of ATRA until complete remission (CR) was achieved, defined as: a) presence of less than 5% blasts in the bone marrow, with b) white blood cells >103/mm3, c) platelets >105/mm3 and d) hemoglobin concentration >8 g/dl, with no blood or platelet transfusions. Thirty-one (83.7%) patients achieved CR by day 50, and 75% of these before day 30. Correction of the coagulopathy, achieved between days 2 and 10 (mean, 3 days), was the first evidence of response to treatment. Only one patient had been previously treated with chemotherapy and three had the microgranular variant M3 form. Dryness of skin and mucosae was the most common side effect observed in 82% of the patients. Thrombosis, hepatotoxicity and retinoid acid syndrome (RAS) were observed in 7 (19%), 6 (16%) and 4 (11%) patients, respectively. Thirteen (35%) patients had to be submitted to chemotherapy due to hyperleukocytosis (above 40 x 103/mm3) and six of these presented with new signs of coagulopathy after chemotherapy. Four (11%) patients died secondarily to intracerebral hemorrhage (IH) and two (5.4%) dropped out of the protocol due to severe ATRA side effects (one RAS and one hepatotoxicity). RAS and IH were related strictly to hyperleukocytosis. The reduced use of platelets and fresh frozen plasma probably lowered the total cost of treatment. We conclude that ATRA is an effective agent for inducing complete remission in APL patients.

Incorporation of dietary trans monounsaturated fatty acids into tissues of Walker 256 tumor-bearing rats

The correlation between dietary trans fatty acids and neoplasia was examined in the present study. Walker 256 tumor-bearing and control rats were fed a trans monounsaturated fatty acid (MUFA)-rich diet for 8 weeks and the incorporation of trans fatty acids by tumor tissue was examined. Also, the effect of tumor growth on trans fatty acid composition of plasma and liver, and the content of thiobarbituric acid-reactive substances (TBARS) was determined. Walker 256 tumor cells presented both trans and cis MUFAs given in the diet. The equivalent diet proportions were 0.66 for trans and 1.14 for cis. Taking into consideration the proportion of trans MUFAs in plasma (11.47%), the tumor incorporated these fatty acids in a more efficient manner (18.27%) than the liver (9.34%). Therefore, the dietary trans fatty acids present in the diet are actively incorporated by the tumor. Tumor growth itself caused marked changes in the proportion of polyunsaturated fatty acids in the plasma and liver but provoked only slight modifications in both trans and cis MUFAs. Tumor growth also reduced the unsaturation index in both plasma and liver, from 97.79 to 86.83 and from 77.51 to 69.64, respectively. This effect was partially related to an increase in the occurrence of the lipid oxidation/peroxidation process of TBARS content which was increased in both plasma (from 0.428 to 0.505) and liver (from 9.425 to 127.792) due to tumor growth.

The non-palindromic adaptor-PCR method for the identification of the T-cell receptor genes of an interferon-gamma-secreting T-cell hybridomaspecific for trans-sialidase, an immunodominant Trypanosoma cruzi antigen

Cloning of the T-cell receptor genes is a critical step when generating T-cell receptor transgenic mice. Because T-cell receptor molecules are clonotypical, isolation of their genes requires reverse transcriptase-assisted PCR using primers specific for each different Valpha or Vß genes or by the screening of cDNA libraries generated from RNA obtained from each individual T-cell clone. Although feasible, these approaches are laborious and costly. The aim of the present study was to test the application of the non-palindromic adaptor-PCR method as an alternative to isolate the genes encoding the T-cell receptor of an antigen-specific T-cell hybridoma. For this purpose, we established hybridomas specific for trans-sialidase, an immunodominant Trypanosoma cruzi antigen. These T-cell hybridomas were characterized with regard to their ability to secrete interferon-gamma, IL-4, and IL-10 after stimulation with the antigen. A CD3+, CD4+, CD8- interferon-gamma-producing hybridoma was selected for the identification of the variable regions of the T-cell receptor by the non-palindromic adaptor-PCR method. Using this methodology, we were able to rapidly and efficiently determine the variable regions of both T-cell receptor chains. The results obtained by the non-palindromic adaptor-PCR method were confirmed by the isolation and sequencing of the complete cDNA genes and by the recognition with a specific antibody against the T-cell receptor variable ß chain. We conclude that the non-palindromic adaptor-PCR method can be a valuable tool for the identification of the T-cell receptor transcripts of T-cell hybridomas and may facilitate the generation of T-cell receptor transgenic mice.

Avaliação do consumo e análise da rotulagem nutricional de alimentos com alto teor de ácidos graxos trans

Nas últimas décadas, diversos estudos vêm sendo realizados visando avaliar os efeitos dos ácidos graxos trans sobre o organismo e identificar seu mecanismo de ação. Entretanto, somente a cerca de um ano, este item foi incluído na rotulagem nutricional obrigatória brasileira, permitindo ao consumidor controlar o consumo de ácidos graxos trans. Assim, o objetivo deste estudo foi avaliar a adequação de alguns alimentos com alto teor de ácidos graxos trans (biscoitos, sorvetes, chocolates e fast-food) frente à legislação pertinente e, ainda, o consumo diário por adultos e crianças observando a recomendação da OMS. A avaliação da rotulagem nutricional demonstrou que a maioria das amostras analisadas ainda não se adequou à nova legislação. Com base na análise dos questionários de consumo, identificou-se que 39,7% dos adultos e 41,4% das crianças consomem, diariamente, pelo menos um alimento com alto teor de ácidos graxos trans. Observou-se ainda, através do consumo estimado, que a ingestão parcial ou total destes produtos ultrapassa a recomendação diária para adultos (2 g) e crianças (1 ano-0,8 g e 10 anos-1,9 g). Assim, uma atuação efetiva dos órgãos de fiscalização e a promoção de ações educativas visando à menor utilização desses produtos na alimentação deveriam ser estimuladas.

Role of Phosphorylation of the Oxysterol Binding Protein (OSBP) in (PI(4)P) and Sterol-Containing Membrane Recognition and Binding

Studies have demonstrated that the oxysterol binding protein (OSBP) acts as a phosphatidylinositol phosphate (PIP)-sterol exchanger at membrane contact sites (MCS) of the endoplasmic reticulum (ER) and Golgi. OSBP is known to pick up phosphatidylinositol-4-phosphate (PI(4)P) from the ER, transfer it to the trans-Golgi in exchange for a cholesterol molecule that is then transferred from the trans-Golgi to the ER. Upon further examination of this pathway by Ridgway et al. (1), it appeared that phosphorylation of OSBP played a role in the localization of OSBP. The dephosphorylation state of OSBP was linked to Golgi localization and the depletion of cholesterol at the ER. To mimic the phosphorylated state of OSBP, the mutant OSBP-S5E was designed by Ridgway et al. (1). The lipid and sterol recognition by wt-OSBP and its phosphomimic mutant OSBP-S5E were investigated using immobilized lipid bilayers and dual polarization interferometry (DPI). DPI is a technique in which the protein binding affinity to immobilized lipid bilayers is measured and the binding behavior is examined through real time. Lipid bilayers containing 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) and varying concentrations of PI(4)Ps or sterols (cholesterol or 25-hydroxycholesterol) were immobilized on a silicon nitride chip. It was determined that wt-OSBP binds differently to PI(4)P-containing bilayers compared to OSBP-S5E. The binding behavior suggested that wt-OSBP extracts PI(4)P and the change in the binding behavior, in the case of OSBP-S5E, suggested that the phosphorylation of OSBP may prevent the recognition and/or extraction of PI(4)P. In the presence of sterols, the overall binding behavior of OSBP, regardless of phosphorylation state, was fairly similar. The maximum specific bound mass of OSBP to sterols did not differ as the concentration of sterols increased. However, comparing the maximum specific bound mass of OSBP to cholesterol with oxysterol (25-hydroxycholesterol), OSBP displayed nearly a 2-fold increase in bound mass. With the absence of the wt-OSBP-PI(4)P binding behavior, it can be speculated that the sterols were not extracted. In addition, the binding behavior of OSBP was further tested using a fluorescence based binding assay. Using 22-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)-23,24-bisnor-5-cholen-3β-ol (22-NBD cholesterol), wt-OSBP a one site binding dissociation constant Kd, of 15 ± 1.4 nM was determined. OSBP-S5E did not bind to 22-NBD cholesterol and Kd value was not obtained.