Rapid Determination of Diuretics in Human Urine by Gas Chromatography – Mass Spectrometry Following Microwave Assisted Derivatization

This work presents a GC–MS–MS–MS method for the direct determination of clenbuterol in human urine. The method 3 comprises a pretreatment procedure and the instrumental analysis of the derivatives performed by GC–MS (ion trap) with 3 electron impact ionization. The GC–MS analysis allows isolation and characterization of specific fragments from the 1 original (MS ) molecular structure, and in particular, those fragments originating from the precursor ion cluster (m/z5335– 2 337) characteristic of clenbuterol. The MS product fragment m/z5300 is in turn used as a further precursor fragment 3 4 giving rise to a MS spectrum specific for clenbuterol. MS fragmentation spectra were also investigated. However, further 3 4 fragmentation of MS product ions does not lead to functional MS spectra nor to any significant increase in the 3 signal-to-noise ratio. The sensitivity limit of the MS technique is lower than 0.2 mg/ l, with a linear range between 0.5 and 5 mg/ l, thus matching the basic requirements for antidoping analysis according to the guidelines of the International Olympic Committee. Due to its overall analytical performance, the method is presently being evaluated as a confirmation protocol to be followed to detect illicit clenbuterol administration to the athletes, and compared with reference GC–MS and GC–MS–MS techniques

Cephalosporin-induced Hemolytic Anemia in a Sicilian Child.

A 27-month-old child developed acute hemolysis on two occasions after the administration of cephalosporin. On the first occasion, hemolysis was intravascular and was due to the formation of complexes between antibodies and the drug, which bound to red blood cells and caused severe hemolysis. On the second occasion, hemolysis was extravascular and was probably due to antibody-dependent cell mediated cytotoxicity. Marked increases in levels of CD19(+), and CD57(+) CD8(+) cells were detected among the subpopulations of the patient's lymphocytes but only in the level of CD19(+) cells from the patient's father, after incubation of a sample of whole blood with a solution of cephalosporins. These results might explain the differences between the immune response of the patient and those of other members of his family and of an unrelated control

Curcumin-loaded polymeric and lipid nanocapsules: preparation, characterization and chemical stability evaluation

Polymeric and lipid nanocapsules suspensions of the natural compound curcumin were prepared in order to overcome limitations associated with its clinical applications, such as poor aqueous solubility and susceptibility to hydrolytic and photochemical degradation. Nanocapsule suspensions were prepared by nanoprecipitation and phase inversion methods, respectively. The curcumin formulations were investigated for physicochemical characteristics and in vitro drug release. The hydrolytic and photochemical degradation of the drug associated with the nanocarriers was also determined. For all formulations, the entrapment efficiency values were higher than 99 %. The aqueous colloidal suspensions of curcumin resulted in an increase in drug concentration by a factor of up to 46.103 times. Moreover, stability studies indicated that nanoencapsulation slows down the hydrolytic and photochemical degradations of curcumin. The strategy of nanoencapsulation into polymeric and lipid nanocapsules produced a formulation of curcumin with high drug loading and improved stability, representing a good strategy for the delivery of this drug.Colegio de Farmacéuticos de la Provincia de Buenos Aire

Defective Function of the Fas Apoptotic Pathway in Type 1 Diabetes Mellitus Correlates with Age at Onset

The Fas death receptor triggers lymphocyte apoptosis through an extrinsic and an intrinsic pathway involving caspase-8 and -9 respectively. Inherited defects of Fas function are displayed by a proportion of patients with Type 1 diabetes mellitus (T1DM) especially those with a second autoimmunity (T1DM-p). This study assesses activation of both pathways in Fas-resistant (FasR) patients to localize the defect. 21/28 (75%) T1DM-p, 14/50 (38%) T1DM, and 7/150 (5%) controls were FasR. Analysis of the 35 FasR patients and 20 Fas-sensitive (FasS) controls showed that caspase-9 activity was lower in T1DM-p and T1DM than in controls, whereas caspase-8 activity was lower in T1DM-p than in T1DM and the controls. Single patient analysis showed that 16/35 patients displayed defective activity of one (FasR1), whereas 19 displayed normal activity of both caspases (FasR2) Ages at onset of diabetes mellitus in T1DM and the second autoimmune disease in T1DM-p were lower in FasR than in FasS patients. All FasR1 patients developed diabetes mellitus before the age of 9 years, whereas a later onset was displayed by 26% FasR2 and 53% FasS patients. These data show that defective Fas function may involve both the extrinsic and intrinsic pathway in T1DM and severity correlates with the precocity of the autoimmune attack and its tissue polyreactivity

ESTUDO DA MINERALlZAÇÃO DO NITROGÊNIO DA MATÉRIA ORGÂNICA DO SOLO SOB SISTEMAS AGROFLORESTAIS DE CORDIA E ERYTHRINA EM TURRIALBA, COSTA RICA

Estudou-se, durante o período de novembro/89 a junho/90, a mineralização do nitrogênio da matériaorgânicado solo. O estudo abrangeu as parcelas agroflorestais com cultivos perenes: café (Coffea arabica) com árvore leguminosa de sombra, poró (Erythrina poeppigiana); café com árvore madeirável de sombra, laurel (Cordia alliodora); cacau (Theobroma cacao) com poró e cacau com laurel, num desenho de blocos completos ao acaso, com parcelas divididas ("split plot", café ou cacau). Observou-se que ao término de 30 dias todo o amônio inicial existente nos sistemas é transformado em nitratos. Foi observada uma relação inversa entre a umidade do solo e a mineralização. No entanto o efeito da umidade foi menos marcado que o do tipo de vegetação, uma vez que os sistemas com poró apresentaram a maior taxa de mineralização, apesar de que a umidade no solo foi também maior que nos sistemas com laurel. A quantidade de nitrogênio disponível para as plantas, remanescente no solo, também foi maior nos sistemas com poró

High levels of osteopontin associated with polymorphisms in its gene are a risk factor for development of autoimmunity/lymphoproliferation

The autoimmune/lymphoproliferative syndrome (ALPS) displays defective function of Fas, autoimmunities, lymphadenopathy/splenomegaly, and expansion of CD4/CD8 double-negative (DN) T cells. Dianzani autoimmune/lymphoproliferative disease (DALD) is an ALPS variant lacking DN cells. Both forms have been ascribed to inherited mutations hitting the Fas system but other factors may be involved. A pilot cDNA array analysis on a DALD patient detected overexpression of the cytokine osteopontin (OPN). This observation was confirmed by enzyme-linked immunosorbent assay (ELISA) detection of higher OPN serum levels in DALD patients (n = 25) than in controls (n = 50). Analysis of the OPN cDNA identified 4 polymorphisms forming 3 haplotypes (A, B, and C). Their overall distribution and genotypic combinations were different in patients (N = 26) and controls (N = 158) (P <.01). Subjects carrying haplotype B and/or C had an 8-fold higher risk of developing DALD than haplotype A homozygotes. Several data suggest that these haplotypes influence OPN levels: (1) in DALD families, high levels cosegregated with haplotype B or C; (2) in healthy controls, haplotype B or C carriers displayed higher levels than haplotype A homozygotes; and (3) in AB and AC heterozygotes, mRNA for haplotype B or C was more abundant than that for haplotype A. In vitro, exogenous OPN decreased activation-induced T-cell death, which suggests that high OPN levels are involved in the apoptosis defect

Curcumin-loaded lipid and polymeric nanocapsules stabilized by nonionic surfactants: An In Vitro and In Vivo antitumor activity on B16-F10 melanoma and macrophage uptake comparative study

Curcumin is a polyphenol obtained from the plant Curcuma longa (called turmeric) that displays several pharmacological activities, including anti-inflammatory, antioxidant, antimicrobial and antitumoral activity, but clinical use has been limited by its poor solubility in water and, consequently, minimal systemic bioavailability. We have therefore formulated the drug into nanocarrier systems in an attempt to improve its therapeutic properties. This study evaluates the effect of intraperitoneally administered nanocapsules containing curcumin on subcutaneous melanoma in mice inoculated with B16-F10 cells, and on the cytotoxicity activity against B16-F10 cells in vitro. Phagocytic uptake of formulations was also evaluated upon incubation with macrophage J774 cells by fluorescence microscopy. Lipid and polymeric nanocapsules were prepared by the phase inversion and nanoprecipitation methods, respectively. The uptake of the lipid nanocapsules prepared using Solutol HS15 was significantly reduced in J774 cells. Curcumin, as free drug or as drug-loaded nanocapsules, was administrated at a dose of 6 mg/kg twice a week for 21 days. Free drug and curcuminloaded nanocapsules significantly reduced tumor volume (P < 0.05 vs. control), but no difference was found in the antitumor activity displayed by lipid and polymeric nanocapsules. This assumption was supported by the in vitro study, in which free curcumin as well as loaded into nanocapsules caused significant reduction of cell viability in a concentration- and time-dependent manner.Fil: Mazzarino, Letícia. Universidade Federal de Santa Catarina; BrasilFil: Silva, Luís F. C.. Universidade Federal de Santa Catarina; BrasilFil: Curta, Juliana C.. Universidade Federal de Santa Catarina; BrasilFil: Licínio, Marley A.. Universidade Federal de Santa Catarina; BrasilFil: Costa, Aline. Universidade Federal de Santa Catarina; BrasilFil: Pacheco, Letícia K.. Universidade Federal de Santa Catarina; BrasilFil: Siqueira, Jarbas M.. Universidade Federal de Santa Catarina; BrasilFil: Martinetti Montanari, Jorge Anibal. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Romero, Eder. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología; ArgentinaFil: Assreuy, Jamil. Universidade Federal de Santa Catarina; BrasilFil: Santos Silva, Maria C.. Universidade Federal de Santa Catarina; BrasilFil: Lemos Senna, Elenara. Universidade Federal de Santa Catarina; Brasi

Detection of bcl-2 rearrangement in mucosa-associated lymphoid tissue lymphomas from patients with hepatitis C virus infection

Lette

Curcumin-loaded polymeric and lipid nanocapsules: preparation, characterization and chemical stability evaluation

Polymeric and lipid nanocapsules suspensions of the natural compound curcumin were prepared in order to overcome limitations associated with its clinical applications, such as poor aqueous solubility and susceptibility to hydrolytic and photochemical degradation. Nanocapsule suspensions were prepared by nanoprecipitation and phase inversion methods, respectively. The curcumin formulations were investigated for physicochemical characteristics and in vitro drug release. The hydrolytic and photochemical degradation of the drug associated with the nanocarriers was also determined. For all formulations, the entrapment efficiency values were higher than 99 %. The aqueous colloidal suspensions of curcumin resulted in an increase in drug concentration by a factor of up to 46.103 times. Moreover, stability studies indicated that nanoencapsulation slows down the hydrolytic and photochemical degradations of curcumin. The strategy of nanoencapsulation into polymeric and lipid nanocapsules produced a formulation of curcumin with high drug loading and improved stability, representing a good strategy for the delivery of this drug.Colegio de Farmacéuticos de la Provincia de Buenos Aire

Roles of the Raf/MEK/ERK and PI3K/PTEN/Akt/mTOR pathways in controlling growth and sensitivity to therapy-implications for cancer and aging

Dysregulated signaling through the Ras/Raf/MEK/ERK and PI3K/PTEN/Akt/mTOR pathways is often the result of genetic alterations in critical components in these pathways or upstream activators. Unrestricted cellular proliferation and decreased sensitivity to apoptotic-inducing agents are typically associated with activation of these pro-survival pathways. This review discusses the functions these pathways have in normal and neoplastic tissue growth and how they contribute to resistance to apoptotic stimuli. Crosstalk and commonly identified mutations that occur within these pathways that contribute to abnormal activation and cancer growth will also be addressed. Finally the recently described roles of these pathways in cancer stem cells, cellular senescence and aging will be evaluated. Controlling the expression of these pathways could ameliorate human health