Mature dendritic cells induce T-helper type-1-dominant immune responses in patients with metastatic renal cell carcinoma

We performed a pilot study on a dendritic cell (DC)-based vaccine in 4 patients with advanced renal cell carcinoma. The vaccine consisted of cultured blood DCs loaded with autologous tumor cell lysate plus keyhole limpet hemocyanin (KLH) and matured with a combination of tumor necrosis factor alpha and prostaglandin E(2). We describe the immune response against KLH induced by DC-based immunization in a patient undergoing an objective partial response and compare it with the responses observed in patients with either stable or progressive disease. The patient with the clinical response developed strong delayed-type hypersensitivity (DTH) against KLH after a single vaccination with antigen-loaded DCs, whereas the other patients failed to develop DTH reactivity even after repeated vaccinations. Antigenic stimulation of mononuclear cells (MNCs) induced proliferation and IFN-gamma but not IL-4 production as well as expression of the chemokine receptor CXCR3 consistent with a T-helper (Th) type-1 bias. Exogenous IL-12 enhanced and exogenous IL-4 diminished IFN-gamma production. In the 2 patients with stable disease two or more vaccinations were required to induce maximal MNC responses. In the patient with progressive disease MNC responses were hardly detectable. Anti-KLH antibodies appeared with different kinetics but could be detected in the serum of all patients. Isotype analysis revealed the presence of IgM, IgG(1), IgG(2) and IgG(3) as well as IgA and complete absence of IgE. The patient with progressive disease also developed IgG(4) antibodies indicative of a deviation towards Th2. Cultured blood DCs can be a potent vaccine for the antigen-specific immunization of patients with advanced kidney cancer. KLH serves as a tracer molecule which allows determination of the magnitude, kinetics and Th bias of the cellular and humoral immune response induced by DC-based immunization. The data also suggest that Th type-1-dominant immune responses involving DTH reaction are required for the induction of tumor regression.     

A static,closed and scaffold-free bioreactor system that permits chondrogenesis in vitro

OBJECTIVE: To characterise in vitro engineered cartilaginous constructs made employing a novel static, scaffold-free and closed chamber system. DESIGN: Chondrocytes derived from slaughter age pigs (3-6 months) were seeded at high density (20 x 10(6)) into cylindrical chambers (1.0 x 0.5cm) and were maintained between an upper and a lower membrane (100 kDa) for 21 days and subsequently cultured in open culture for 7 additional days. RESULTS: Viable constructs produced were approximately 10 mmx2mm in size and were stable enough to be handled by surgical pincers. Histology and electron microscopy evaluations revealed a neo-cartilage structure of high cell density with a comprehensive extracellular matrix. Predominately collagen type II and negligible amounts of collagen types I and X were detected using RT-PCR and SDS-PAGE analyses. CONCLUSIONS: In this study, we provide evidence of a scaffold-free system that can produce immature hyaline-like cartilaginous constructs suitable for in vivo implantation, or that may be useful for in vitro studies of events related to the process of chondrogenesis.     

Lack of a clinically significant pharmacokinetic interaction between fenofibrate and pravastatin in healthy volunteers

This study was conducted to evaluate the potential pharmacokinetic interaction between fenofibrate and pravastatin. A total of 23 healthy adult volunteers received single-dose 201 mg fenofibrate alone, 201 mg fenofibrate + 40 mg pravastatin, and 40 mg pravastatin alone in a three-period crossover experiment. Plasma samples were collected at predetermined times and were analyzed with validated methods for the quantitation of fenofibric acid, pravastatin, and 3 alpha-hydroxy-isopravastatin (3 alpha-iso-PV). Pharmacokinetic parameters of these three compounds were calculated using noncompartmental methods and compared by analyses of variance and bioavailability assessments. Concomitant administration of fenofibrate and pravastatin did not affect the pharmacokinetics of either fenofibric acid or pravastatin. However, the AUC0-infinity and Cmax of 3 alpha-iso-PV were increased by 26% and 29%, respectively. The moderate increase in the formation of this pravastatin metabolite should not raise any clinical concerns due to its much lower pharmacological potency compared to pravastatin and lack of toxicity.     

Insulin receptors and membrane transport in muscle

Summary The AA. report a group ofin vitro experiments relative to the direct inhibition of the insulin effect on the transport of 3-o-methyl glucose and L-proline in rat diaphragm muscle by a reagent specific for tryptophan in a neutral solution, the 2-hydroxy-5-nitrobenzyl bromide. Evidence for the presence of tryptophan at an insulin receptor site in the plasma membrane is given.

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Zusammenfassung Die AA. berichten ueber eine Serie vonin vitro Untersuchungen betr. die direkte Hemmung des Insulineffektes auf den aktiven Transport der 3-o-methyl-Glukose und des l-Prolins beim Rattenzwerchfell mittels eines spezifischen Reagenz' fuer das Tryptophan in neutraler Loesung, des 2-hydroxy-5-nitrobenzil-bromid. Somit ist die Gegenwart von Tryptophan in einem Insulinrezeptor der Plasmamembran bewiesen.

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Resumen Los AA. describen un grupo de experimentosin vitro concernientes a la inhibición directa del efecto de la insulina en el transporte activo de la 3-0-metil glucosa y de la l-prolina en el diafragma de rata por parte de un reactivo específico para el triptófano en solución neutra, el 2-hidroxi-5-nitrobenzil bromuro. De esta manera se comprueba la presencia de triptófano en un receptor insulínico de la membrana plasmática.

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Resume Les AA. rapportent un groupe d'épreuvesin vitro au sujet de l'inhibition directe de l'effet de l'insuline sur le transport actif des 3-o-metil glucose et de la l-proline dans le diaphragme de rat par un réactif spécifique pour le triptophane en solution neutre, le 2-idrossi-5-nitrobenzil bromure. On prouve ainsi la présence de triptophane dans un récepteur insulinique de la membrane plasmatique.

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Riassunto Gli AA. riferiscono un gruppo di esperimentiin vitro riguardanti l'inibizione diretta dell'effetto dell'insulina sul trasporto attivo del 3-o-metil glucosio e della l-prolina nel diaframma di ratto da parte di un reagente specifico per il triptofano in soluzione neutra, il 2-idrossi-5-nitrobenzil bromuro. Viene così provata la presenza di triptofano in un recettore insulinico della membrana plasmatica.

     

Long-term follow-up of the Moberg key grip procedure

The long-term results of the key grip procedure (tenodesis of the flexor pollicis longus tendon to the radius, release of the A1 pulley, and percutaneous pin fixation of the interphalangeal joint of the thumb) were evaluated in 10 tetraplegic patients. Seven patients also had tenodesis of the extensor pollicis longus and brevis tendons to prevent flexion at the metacarpophalangeal (MP) joint. Patients were examined an average of 7.4 years after surgery. Progressive flexion of the MP joint gradually occurred, indicating failure of the extensor tenodesis. Excessive bowstringing of the flexor pollicis longus tendon across the MP joint occurred in nine patients. Because of failure of the extensor tenodesis and bowstringing of the flexor tendon, the patients had to progressively extend the wrist further to pinch small objects. Functional testing demonstrated improved hand use in eight patients. Lateral pinch strength was related to wrist extension torque.