The p85 subunit of phosphatidylinositol 3-kinase (PI3K) has long been thought of as a regulatory subunit that has no other function than the regulation of the p110 catalytic subunit. Our laboratory is studying other roles of the p85 subunit, in particular determining the role of the p85 BH domain. The BH domain has homology to GTPase activating protein (GAP) domains that are involved in the stimulation of monomeric G proteins to hydrolyze their bound GTP to GDP. This converts the G protein from its active conformation to its inactive conformation. We have determined that p85 interacts with Rab proteins, monomeric G proteins that regulate vesicle fusion during the endocytosis of receptors. We have shown that p85 binds to Rab5 regardless of nucleotide-bound state of Rab5. The p85 subunit of PI3K has in vitro GAP activity towards Rab5. It was determined that p85 also has in vitro GAP activity towards Rab4, Rab7, Rab6 as well as the Rho-family G proteins, Rac1 and Cdc42. This GAP activity was localized to the BH domain of p85 and mutation of Arg 274 to Ala abolishes the GAP activity of p85. When this p85R274A mutant was expressed in cells, PDGFR degradation was severely inhibited and there was a corresponding increase in the duration of MAPK and Akt signalling. This increase in cell signalling caused a transformed phenotype in cells expressing the p85 protein with the Arg 274 mutation. These cells have lost contact inhibition for growth, are able to grow independent of attachment as well as in the presence of limited growth factors. They also form tumours in nude mice. These cellular effects seem to be due to an increase in receptor recycling because of the loss of the GAP activity of p85. This increase in receptor recycling may interfere with receptor targeting to the late endosome, which would cause the decrease in receptor degradation that is seen in the p85R274A cells.

The p85 subunit of phosphatidylinositol 3-kinase (PI3K) has long been thought of as a regulatory subunit that has no other function than the regulation of the p110 catalytic subunit. Our laboratory is studying other roles of the p85 subunit, in particular determining the role of the p85 BH domain. The BH domain has homology to GTPase activating protein (GAP) domains that are involved in the stimulation of monomeric G proteins to hydrolyze their bound GTP to GDP. This converts the G protein from its active conformation to its inactive conformation. We have determined that p85 interacts with Rab proteins, monomeric G proteins that regulate vesicle fusion during the endocytosis of receptors. We have shown that p85 binds to Rab5 regardless of nucleotide-bound state of Rab5. The p85 subunit of PI3K has in vitro GAP activity towards Rab5. It was determined that p85 also has in vitro GAP activity towards Rab4, Rab7, Rab6 as well as the Rho-family G proteins, Rac1 and Cdc42. This GAP activity was localized to the BH domain of p85 and mutation of Arg 274 to Ala abolishes the GAP activity of p85. When this p85R274A mutant was expressed in cells, PDGFR degradation was severely inhibited and there was a corresponding increase in the duration of MAPK and Akt signalling. This increase in cell signalling caused a transformed phenotype in cells expressing the p85 protein with the Arg 274 mutation. These cells have lost contact inhibition for growth, are able to grow independent of attachment as well as in the presence of limited growth factors. They also form tumours in nude mice. These cellular effects seem to be due to an increase in receptor recycling because of the loss of the GAP activity of p85. This increase in receptor recycling may interfere with receptor targeting to the late endosome, which would cause the decrease in receptor degradation that is seen in the p85R274A cells.

Growth and proliferation are normal functions of cells mediated in part via receptor tyrosine kinases such as the epidermal growth factor (EGF) receptor (EGFR). The EGFR binds the extracellular signaling ligand EGF and transduces the signal into the cell. Phosphatidylinositol 3'-kinase (PI3K) responds to EGFR activation and initiates downstream signaling cascades responsible for cell cycle entry, proliferation and inhibition of apoptosis. Cell cycle arrest is required to stop cell growth and proliferation as well as allow apoptosis, if required. The phosphatase and tensin homologue deleted on chromosome ten (PTEN) directly opposes PI3K signaling since its substrate is the PI3K product phosphatidylinositol 3,4,5-trisphosphate. PI3K is a heterodimer composed of a p85 regulatory subunit and a p110 catalytic subunit. PTEN is an essential tumor suppressor protein. Absence of PTEN has been associated with several types of cancer. <p>Our laboratory has characterized new specialized functions for the p85 protein. One function discovered was the ability of p85 to enhance PTEN lipid phosphatase activity. In this thesis PTEN activity is shown to be enhanced at least 3.5 fold in vitro by an equimolar amount of p85. <p>We performed an analysis of PTEN using seven PTEN mutants. Two types of mutants were created: i) regulatory or possible regulatory phosphorylation sites were substituted to mimic both phosphorylated and non-phosphorylated states and ii) alanine substitution of basic amino acid residues. The phosphorylation sites altered were the casein kinase 2 phosphorylation sites in the regulatory domain and tyrosine 336, a proposed regulatory phosphorylation site. Three mutants involving alanine substitution for basic amino acid residues included one mutant in the PASE domain and two more mutants in the C2 domain. It was observed that GFP-PTEN translocates to the plasma membrane upon EGF stimulation. The mimic of constitutive phosphorylation of the Casein kinase 2 sites resulted in cytoplasmic localization whereas the non-phosphorylated mimic was plasma membrane localized regardless of EGFR activation status. Neutralization of positive charge in the PASE and C2 domains seriously impeded the ability of PTEN to bind to phosphorylated phosphatidylinositol lipids and abolished the ability of the protein to translocate to the plasma membrane in response to receptor activation. Located within a cluster of positively charged lysine residues in the C2 domain is a potential phosphorylation site at tyrosine 336. The phosphorylation mimic showed decreased binding to some membrane lipids compared to the non-phosphorylated mimic. The results we generated are consistent with a current model for PTEN regulation that proposes PTEN is localized to the cytoplasm in quiescent cells and dephosphorylation of the regulatory domain occurs upon EGF stimulation allowing translocation to the plasma membrane. The model proposes that dephosphorylation of the casein kinase 2 sites unmasks regions of positive charge that interact with the anionic plasma membrane. Furthermore, the results suggested that at the plasma membrane p85 interacts with PTEN to increase lipid phosphatase activity and may be involved in targeting PTEN to the activated receptor where PI3,4,5P3 lipids are being produced.

Growth and proliferation are normal functions of cells mediated in part via receptor tyrosine kinases such as the epidermal growth factor (EGF) receptor (EGFR). The EGFR binds the extracellular signaling ligand EGF and transduces the signal into the cell. Phosphatidylinositol 3'-kinase (PI3K) responds to EGFR activation and initiates downstream signaling cascades responsible for cell cycle entry, proliferation and inhibition of apoptosis. Cell cycle arrest is required to stop cell growth and proliferation as well as allow apoptosis, if required. The phosphatase and tensin homologue deleted on chromosome ten (PTEN) directly opposes PI3K signaling since its substrate is the PI3K product phosphatidylinositol 3,4,5-trisphosphate. PI3K is a heterodimer composed of a p85 regulatory subunit and a p110 catalytic subunit. PTEN is an essential tumor suppressor protein. Absence of PTEN has been associated with several types of cancer. <p>Our laboratory has characterized new specialized functions for the p85 protein. One function discovered was the ability of p85 to enhance PTEN lipid phosphatase activity. In this thesis PTEN activity is shown to be enhanced at least 3.5 fold in vitro by an equimolar amount of p85. <p>We performed an analysis of PTEN using seven PTEN mutants. Two types of mutants were created: i) regulatory or possible regulatory phosphorylation sites were substituted to mimic both phosphorylated and non-phosphorylated states and ii) alanine substitution of basic amino acid residues. The phosphorylation sites altered were the casein kinase 2 phosphorylation sites in the regulatory domain and tyrosine 336, a proposed regulatory phosphorylation site. Three mutants involving alanine substitution for basic amino acid residues included one mutant in the PASE domain and two more mutants in the C2 domain. It was observed that GFP-PTEN translocates to the plasma membrane upon EGF stimulation. The mimic of constitutive phosphorylation of the Casein kinase 2 sites resulted in cytoplasmic localization whereas the non-phosphorylated mimic was plasma membrane localized regardless of EGFR activation status. Neutralization of positive charge in the PASE and C2 domains seriously impeded the ability of PTEN to bind to phosphorylated phosphatidylinositol lipids and abolished the ability of the protein to translocate to the plasma membrane in response to receptor activation. Located within a cluster of positively charged lysine residues in the C2 domain is a potential phosphorylation site at tyrosine 336. The phosphorylation mimic showed decreased binding to some membrane lipids compared to the non-phosphorylated mimic. The results we generated are consistent with a current model for PTEN regulation that proposes PTEN is localized to the cytoplasm in quiescent cells and dephosphorylation of the regulatory domain occurs upon EGF stimulation allowing translocation to the plasma membrane. The model proposes that dephosphorylation of the casein kinase 2 sites unmasks regions of positive charge that interact with the anionic plasma membrane. Furthermore, the results suggested that at the plasma membrane p85 interacts with PTEN to increase lipid phosphatase activity and may be involved in targeting PTEN to the activated receptor where PI3,4,5P3 lipids are being produced.

Rab5 (Ras-related in brain) and Rab4 are small monomeric GTPases that mediate the intracellular trafficking of endocytosed growth factor receptors. Active Rab5-GTP has low intrinsic GTP hydrolysis activity that is stimulated by GTPase activating proteins (GAPs) to make inactive Rab5-GDP. GAPs provide both a catalytic arginine and switch region stabilization functions. The p85 regulatory subunit of phosphatidylinositol 3′-kinase (PI3K) has GAP activity towards Rab5 and Rab4, which is not seen in other PI3Ks. The arginine “finger” residue within p85 is R274. It is unlikely that p85 stabilizes the switch regions of Rab5, which undergo large conformational changes between activation states, because it interacts with both Rab5-GTP and Rab5-GDP. In contrast, the PI3K catalytic subunit p110β binds only Rab5-GTP, suggesting it interacts with the switch regions. Thus, the GAP functions may be provided to Rab5 by the subunits of PI3K acting together, where p85 provides the arginine finger and p110β stabilizes the switch regions. The binding interface of Rab5:p85 was sought using mutations of Rab5 residues not present in the switch regions which were conserved in p85-binding Rab proteins (S84, E106, N113, F145, E172, M175, K179, K180) in GST pull-down experiments with FLAG-p85. The p85 binding site was not resolved with these experiments, suggesting that p85 interaction may involve the contribution of multiple residues of the Rab5 protein. The p110β interaction site on Rab5 was investigated using Rab5 switch region mutants. Pull-down experiments using a stabilized p110 protein construct, where the p85-iSH2 domain was fused to p110 (alpha or beta), were performed. Rab5 mutants I53A, F57A, W74A, Q79L, E80R, Y82A, H83E, L85A, M88A, Y89A and R91E showed reduced p110β binding. All of these residues except E80 and H83 are involved in binding other Rab5 effectors. The Rab5 binding site on p110β was also resolved through mutation of p110β in its Ras binding domain, and includes residues I234, E238 and Y244. This generation of non-binding mutants of both Rab5 and p110β will be invaluable in the characterization of the importance of the p110β:Rab5-GTP interaction for receptor trafficking to endosomes in mammalian cells.

Prostate cancer is the second leading cause of cancer-related death in men in the Western world. Deregulation of transforming growth factor β (TGFβ) signaling pathway is frequently detected in prostate cancer and contributes to tumor growth, migration, and invasion. In normal tissue and the early stages of cancer, TGFβ acts as a tumor suppressor by regulating proliferation, differentiation, and apoptosis. In later stages of cancer, TGFβ acts as a tumor promoter by inducing angiogenesis, tumor invasion, and migration. Thus, it is important to investigate the molecular mechanisms behind the tumor-promoting effects of TGFβ, which is the topic of this thesis.   The tumor necrosis factor receptor–associated factor 6 (TRAF6) controls non-canonical TGFβ signals due to its enzymatic activity, causing polyubiquitination of the cell membrane–bound, serine/threonine kinase TGFβ type I receptor (TβRI) and its subsequent cleavage in the extracellular domain by tumor necrosis factor a–converting enzyme (TACE) in a protein kinase C ζ (PKCζ)-dependent manner. TRAF6 also recruits the active g-secretase complex to the TβRI, resulting in a second cleavage in the transmembrane region and the liberation of the TβRI intracellular domain (TβRI-ICD), which enters the nucleus, where it associates with the transcriptional co-regulator p300. In Paper I, the aim was to elucidate by which mechanisms TβRI-ICD enters the nucleus. We found that the endocytic adaptor protein APPL1 interacts with TβRI and PKCζ. APPL proteins are required for TβRI translocation from endosomes to the nucleus via microtubules in a TRAF6-dependent manner. Moreover, APPL proteins are important for TGFβ-induced cell invasion, and high levels of APPL1 are detected by immunohistochemistry in prostate cancer. Finally, we demonstrated that the APPL1–TβRI complex visualized with the in situ proximity ligation assay (PLA) correlates with Gleason score, indicating that it might be a novel prognostic marker for aggressive prostate cancer. In Paper II, the aim was to explore by which mechanisms TGFβ causes activation of the AKT pathway, which regulates migration and therapy resistance of cancer cells. We found that the E3 ligase activity of TRAF6 induces Lys63-linked polyubiquitination of p85α upon TGFβ stimulation, resulting in plasma membrane recruitment, Lys63-linked polyubiquitination, and subsequent activation of AKT. Moreover, the TRAF6 and PI3K/AKT pathway were found to be crucial for the TGFβ-induced migration. Importantly, we demonstrated, by PLA, a correlation between Lys63-linked polyubiquitination of p85α and aggressive prostate cancer in tissue sections from patients with prostate cancer. In Paper III, the aim was to investigate the mechanisms for TGFβ-induced activation of PKCζ and the role of PKCζ in tumor regression. We found that TRAF6 caused Lys63-linked polyubiquitination of PKCζ. By using two novel chemical compounds that inhibit PKCζ, we demonstrated that PKCζ is crucial for prostate cancer cell survival and invasion. In Paper IV, the aim was to investigate further the target genes for the nuclear TβRI-ICD-APPL1 complex identified in Paper I. We provide evidence that APPL proteins and the TGFβ signaling pathway are important for cell proliferation. In summary, the results reported in this thesis suggest the potential usefulness of the identified signaling components of the tumor-promoting effects of TGFβ as drug targets and biomarkers for aggressive prostate cancer.

Activated receptor tyrosine kinase (RTK), such as the epidermal growth factor (EGF) receptor (EGFR) and the platelet-derived growth factor (PDGF) receptor (PDGFR), recruit downstream signalling proteins, including phosphatidylinositol 3-kinase (PI3K). PI3K, composed of a regulatory p85 subunit and a catalytic p110 subunit, phosphorylates phosphatidylinositol 4,5-bisphosphate at the 3 position to generate phosphatidylinositol 3,4,5-trisphosphate. This lipid second messenger activates Akt, which promotes cell growth, cell cycle entry and progression, as well as cell survival and cellular migration. PTEN, a tumor suppressor protein, dephosphorylates phosphatidylinositol 3,4,5-trisphosphate at the 3 position, turning off Akt signalling. PTEN contains a C-terminal PDZ binding motif that binds to the PDZ2 domain of MAGI-2, a scaffolding protein that localizes signalling molecules to the plasma membrane. MAGI-2 has ten domains that potentially mediate multiple protein-protein interactions simultaneously. A PTEN associated-complex (PAC) has been described and may contain MAGI-2, PTEN and p85. The PAC is hypothesized to form at the plasma membrane at appropriate sites for PTEN to gain access to its lipid substrates, since the binding of PTEN to MAGI-2 has been shown to enhance the suppression of PI3K-mediated Akt signalling. In order to better understand the role of the PAC in attenuation of the Akt signalling pathway, regions of the MAGI-2 scaffolding protein were mapped to identify the interactions taking place in the PAC. MAGI-2, and its individual domains, were expressed as GST fusion proteins. These were immobilized onto beads and allowed to bind to cellular proteins including PTEN, p85, PDGFR and EGFR using a GST pull-down experiment. The proteins bound to GST-MAGI-2 were identified using an immunoblot analysis. In vitro pull-down experiments revealed that MAGI-2 PDZ2 and PDZ5 domains bind to PTEN, and both MAGI-2 WW domains were shown to bind to p85. EGFR and PDGFR did not bind to the PDZ domains of MAGI-2 under the conditions studied. In order to study protein-protein interactions in cells, immunoprecipitation assays were also performed. Full length MAGI-2 was expressed tagged to a Myc epitope. This was used in immunoprecipitation assays to determine if MAGI-2 could co-immunoprecipitate with proteins involved in the Akt signalling pathway, such as PTEN, p85, PDGFR and EGFR. MAGI-2 can co-immunoprecipitate with PTEN upon 5 min EGF stimulation however, this result was inconclusive because replicate experiments did not verify this initial observation. MAGI-2 does not co-immunoprecipitate with the EGFR nor p85, under the conditions tested. We examined for these interactions after 5 min of growth factor stimulation and more experiments that test different time points after growth factor stimulation would reveal if these interactions are present at shorter time points. MAGI-2 has been shown to bind to PTEN and p85 in vitro and therefore has the potential to regulate the attenuation of the PI3K/Akt signalling pathway in response to activated EGFR and/or PDGFR.

The adaptor subunit of phosphatidylinositol 3'-kinases (PI3K), p85, is involved in many different biological processes. Recent studies have shown that one of these functions is to serve as a GTPase activating protein (GAP) towards Rab5, a small monomeric G-protein. Rab5, like other G-proteins, can bind to either GDP or GTP in vivo, assuming its inactive and active form, respectively. The p85 protein has been shown to associate with both the nucleotide-bound and nucleotide-free states of Rab5. It has also been shown that p85 associates with activated, phosphorylated platelet-derived growth factor receptors (PDGFRs) via its two SH2 domains, and that upon binding there is a conformational change in the p85 protein which leads to a derepression of p110 kinase activity. The purpose of this study was to analyze if binding of the activated PDGFR peptides to p85 affects its Rab5GAP activity, as well as to measure the binding affinity of p85 towards Rab5 in each of its nucleotide-bound states. GAP assays were performed to measure the effect that peptide analogs of both the activated and inactivated PDGFR had on p85 Rab5GAP activity, while the binding affinity of p85 towards Rab5 was measured using surface plasmon resonance. The results of this study suggest that PDGFR peptides have no significant effect on p85 Rab5GAP activity. Furthermore, p85 appears to have a higher magnitude of binding to nucleotide-associated Rab5 proteins, than nucleotide-free Rab5 proteins. It also appears that p85 forms more stable complexes with Rab5-GTP than with Rab5-GDP. These results further support previous studies that show p85 to be an important regulator of Rab5-mediated endosomal fusion and show that this activity is not regulated by binding to the activated PDGFR itself.

Activated receptor tyrosine kinase (RTK), such as the epidermal growth factor (EGF) receptor (EGFR) and the platelet-derived growth factor (PDGF) receptor (PDGFR), recruit downstream signalling proteins, including phosphatidylinositol 3-kinase (PI3K). PI3K, composed of a regulatory p85 subunit and a catalytic p110 subunit, phosphorylates phosphatidylinositol 4,5-bisphosphate at the 3 position to generate phosphatidylinositol 3,4,5-trisphosphate. This lipid second messenger activates Akt, which promotes cell growth, cell cycle entry and progression, as well as cell survival and cellular migration. PTEN, a tumor suppressor protein, dephosphorylates phosphatidylinositol 3,4,5-trisphosphate at the 3 position, turning off Akt signalling. PTEN contains a C-terminal PDZ binding motif that binds to the PDZ2 domain of MAGI-2, a scaffolding protein that localizes signalling molecules to the plasma membrane. MAGI-2 has ten domains that potentially mediate multiple protein-protein interactions simultaneously. A PTEN associated-complex (PAC) has been described and may contain MAGI-2, PTEN and p85. The PAC is hypothesized to form at the plasma membrane at appropriate sites for PTEN to gain access to its lipid substrates, since the binding of PTEN to MAGI-2 has been shown to enhance the suppression of PI3K-mediated Akt signalling. In order to better understand the role of the PAC in attenuation of the Akt signalling pathway, regions of the MAGI-2 scaffolding protein were mapped to identify the interactions taking place in the PAC. MAGI-2, and its individual domains, were expressed as GST fusion proteins. These were immobilized onto beads and allowed to bind to cellular proteins including PTEN, p85, PDGFR and EGFR using a GST pull-down experiment. The proteins bound to GST-MAGI-2 were identified using an immunoblot analysis. In vitro pull-down experiments revealed that MAGI-2 PDZ2 and PDZ5 domains bind to PTEN, and both MAGI-2 WW domains were shown to bind to p85. EGFR and PDGFR did not bind to the PDZ domains of MAGI-2 under the conditions studied. In order to study protein-protein interactions in cells, immunoprecipitation assays were also performed. Full length MAGI-2 was expressed tagged to a Myc epitope. This was used in immunoprecipitation assays to determine if MAGI-2 could co-immunoprecipitate with proteins involved in the Akt signalling pathway, such as PTEN, p85, PDGFR and EGFR. MAGI-2 can co-immunoprecipitate with PTEN upon 5 min EGF stimulation however, this result was inconclusive because replicate experiments did not verify this initial observation. MAGI-2 does not co-immunoprecipitate with the EGFR nor p85, under the conditions tested. We examined for these interactions after 5 min of growth factor stimulation and more experiments that test different time points after growth factor stimulation would reveal if these interactions are present at shorter time points. MAGI-2 has been shown to bind to PTEN and p85 in vitro and therefore has the potential to regulate the attenuation of the PI3K/Akt signalling pathway in response to activated EGFR and/or PDGFR.

Novos tipos de sílicas mesoporosas esféricas (SMEs) com tamanho de partícula entre 3 e 10 &#181;m foram sintetizadas utilizando os copolímeros tribloco EO17PO60EO17 (P103) ou EO26PO39EO26 (P85) como direcionadores de estrutura. As SMEs foram preparadas via um processo de síntese com duas etapas de tratamento hidrotérmico (TH) em forno convencional, utilizando o ortosilicato de tetraetila como fonte de sílica, os surfatantes P103 ou P85 como moldes em combinação com o co-surfatante brometo de cetiltrimetilamônio (CTAB) e o co-solvente etanol, sob condições ácidas. As SMEs obtidas foram caracterizadas por microscopia eletrônica de varredura (MEV) e medidas de adsorção/dessorção de N2. O volume e o tamanho do poro das SMEs podem ser aumentados com o aumento da temperatura do TH. O volume e o tamanho do poro (0,41 cm3g-1; 2,84 nm) da amostra sintetizada com P103 aumentaram (1,20 cm3g-1; 4,32 nm) quando a temperatura do TH aumentou de 80°C para 120°C na segunda etapa do TH. O aumento do volume e do tamanho de poro também pode ser obtido utilizando um único tratamento hidrotérmico, porém empregando 1,3,5-trimetilbenzeno (TMB) como agente dilatador de poros. O volume e o tamanho do poro (0,34 cm3g-1; 2,02 nm) da amostra sintetizada com P85 sem TMB aumentaram (0,37 cm3g-1; 2,51 nm) na amostra preparada com P85 e TMB. As características texturais e de superfície dos materiais obtidos com P103 e P85 foram comparadas com um material sintetizado em paralelo, nas mesmas condições experimentais, utilizando o copolímero EO20PO70EO20 (P123) como direcionador de estrutura e CTAB como co-surfatante. A SME sintetizada com P103 foi usada como adsorvente de compostos orgânicos voláteis (COVs) oriundos de misturas padrões. Os componentes da mistura padrão foram então removidos do adsorvente (sílica) por dessorção térmica e introduzidos em uma coluna cromatográfica para separação por cromatografia a gás (CG) e identificação por espectrometria de massa (EM). Esta SME foi também testada como adsorvente de uma amostra de ar coletada em uma rua com significativo fluxo de veículos. O ar foi coletado paralelamente na SME e num adsorvente comercial (Tenax TA/Carbotrap). Os compostos n-hexano, benzeno, tolueno e o-xileno, oriundos de emissões veiculares, foram encontrados em ambos adsorventes (sílica e Tenax TA/Carbotrap). O fármaco Rifampicina foi encapsulado numa SME sintetizada com P123/CTAB e na sílica SBA-15 (poros ordenados hexagonalmente). A encapsulação do fármaco (cerca de 30%) em ambas as sílicas foi confirmada pelos resultados de adsorção/dessorção de N2. / New types of mesoporous silica spheres with particle diameter of 3 - 10 &#181;m were synthesized by using a triblock copolymer EO17PO60EO17 (P103) or EO26PO39EO26 (P85) as templates. The microspheres were prepared via a two-step hydrothermal treatment (HT) in an oven by using tetraethoxysilane as silica source, the surfactants P103 or P85 as templates in combination with a cosurfactant cetyltrimethylammonium bromide (CTAB) and a cosolvent ethanol, under acidic conditions. The obtained silica spheres in both procedures were characterized by scanning electron microscopy (SEM) and N2 sorption technique. The volume and the pore size of the silica spheres can become greater by increasing the temperature of the HT. The volume and the pore size (0.41 cm3g-1; 2.84 nm) of the sample prepared with P103 became greater (1.20 cm3g-1; 4.32 nm) when the temperature of HT increased by 80°C to 120°C in the second step of the HT. The volume and the pore size can also be increased using 1,3,5-trimethylbenzene (TMB) as a swelling agent, instead of raising the temperature of the HT. The volume and the pore size (0.34 cm3g-1; 2.02 nm) of the sample prepared with P85 without TMB became greater (0.37 cm3g-1; 2.51 nm) in the sample prepared with P85 and TMB. The characteristics of textures and surfaces of the materials synthesized by using P103 or P85 were compared with a material prepared with the copolymer EO20PO70EO20 (P123) as template using the same experimental conditions. The silica microspheres synthesized with P103 were used as adsorbents for volatile organic compounds (VOCs) from standard mixtures. The compounds of the standard mixture were then removed from the adsorbent (silica) by thermal desorption and introduced into a chromatographic column for separation by gas chromatography (GC) and identification by mass spectrometry (MS). This material was also used as adsorbent of an air sample collected on a street with a significant flow of motor vehicles. The air was collected on the silica and on a commercial adsorbent (Tenax TA/Carbotrap) one by one. The compounds n-hexane, benzene, toluene and o-xylene, resulted from the emissions from vehicles, were found in both adsorbents (silica and Tenax TA/Carbotrap). The drug Rifampicin was encapsulated in the mesoporous spherical silica, prepared with P123/CTAB and in the silica SBA-15 (hexagonally ordered pores). The encapsulation of the drug (about 30%) in both the silica was confirmed by measurements of adsorption/desorption of N2