Modulation of cdk2, cyclin D1, p16INK4a, p21WAF and p27Kip1 expression in endothelial cells by TNF/IFN gamma.

BACKGROUND: Regional administration of high doses of tumor necrosis factor (TNF) and interferon gamma (IFN gamma) to metastatic melanoma patients causes selective disruption of the tumor vasculature. This effect is paralleled by decreased endothelial cell proliferation and suppressed integrin alpha V beta 3-mediated adhesion in vitro. Overexpression of the cyclin-dependent kinase (cdk) inhibitory protein p16INK4a was reported to interfere with integrin alpha V beta 3-dependent melanoma cell adhesion. MATERIALS AND METHODS: TNF- and IFN gamma-treated HUVEC were analyzed for cell cycle progression and for protein expression by flow cytometry and Western blotting, respectively. p16INK4a was overexpressed by transient transfection, and HUVEC adhesion was tested in short-term adhesion assays. RESULTS: TNF and IFN gamma synergistically induced a G1 arrest associated with reduced levels of cyclin D1 and cdk2, and increased expression of the cdk inhibitors p16INK4a, p21WAF and p27Kip1. p16INK4a overexpression, however, had no effect on alpha V beta 3-mediated adhesion. CONCLUSION: These results implicate the down-regulation of cyclin D1 and cdk-2, and up-regulation of p16INK4a, p21WAF and p27Kip1 in the suppression of endothelial cell proliferation induced by TNF/IFN gamma and demonstrate that increased p16INK4a levels are not sufficient to suppress alpha V beta 3-mediated endothelial cell adhesion.

Variations in IB1/JIP1 expression regulate susceptibility of beta-cells to cytokine-induced apoptosis irrespective of C-Jun NH2-terminal kinase signaling.

We previously reported that interleukin-1beta (IL-1beta) alone does not cause apoptosis of beta-cells, whereas when combined with gamma-interferon (IFN-gamma) and tumor necrosis factor-alpha (TNF-alpha), it exerts a distinct apoptotic effect. Studies in beta-cell lines indicated that IL-1beta reduced expression of islet brain (IB)-1/JNK interacting protein (JIP)-1, a JNK scaffold protein with antiapoptotic action. We examined whether variations in IB1/JIP-1 expression in purified primary beta-cells affect their susceptibility to cytokine-induced apoptosis. Exposure to IL-1beta for 24 h decreased cellular IB1/JIP-1 content by 66 +/- 17%; this IL-1beta effect was maintained in the presence of TNF-alpha + IFN-gamma, which did not influence IB1/JIP-1 levels by themselves. Addition of IL-1beta to TNF-alpha + IFN-gamma increased apoptosis from 20 +/- 2% to 59 +/- 5%. A similar increase in TNF-alpha + IFN-gamma-induced apoptosis was produced by adenoviral expression of antisense IB1/JIP-1 and was not further enhanced by addition of IL-1beta, indicating that IL-1beta-mediated suppression of IB1/JIP-1 in beta-cells increases their susceptibility to cytokine-induced apoptosis. However, adenovirally mediated overexpression of IB1/JIP-1 also potentiated TNF-alpha + IFN-gamma-induced apoptosis, suggesting that the antiapoptotic effect of IB1/JIP-1 depends on well-defined cellular levels. We conclude that the IB1/JIP-1 level in beta-cells can control their susceptibility to apoptosis independent of JNK signaling.

Vitamin C requirements in parenteral nutrition.

Some biochemical functions of vitamin C make it an essential component of parenteral nutrition (PN) and an important therapeutic supplement in other acute conditions. Ascorbic acid is a strong aqueous antioxidant and is a cofactor for several enzymes. The average body pool of vitamin C is 1.5 g, of which 3%-4% (40-60 mg) is used daily. Steady state is maintained with 60 mg/d in nonsmokers and 140 mg/d in smokers. Shocked surgical, trauma, and septic patients have a drastic reduction of circulating plasma ascorbate concentrations. These low concentrations require 3-g doses/d to restore normal plasma ascorbate concentrations, questioning the recommended PN dose of 100 mg/d. Determination of intravenous requirements is usually based on plasma concentrations, which are altered during the inflammatory response. There is no clear indicator of deficiency: serum or plasma ascorbate concentrations <0.3 mg/dL (20 micromol/L) indicates inadequate vitamin C status. On the basis of available pharmacokinetic data the 100 mg/d dose for patients receiving home PN and 200 mg/d for stable adult patients receiving PN are adequate, but requirements have been shown to be higher in perioperative, trauma, burn, and critically ill patients, paralleling oxidative stress. One recommendation cannot fit all categories of patients. Large vitamin C supplements may be considered in severe critical illness, major trauma, and burns because of increased requirements resulting from oxidative stress and wound healing. Future research should distinguish therapeutic use of high-dose ascorbic acid antioxidant therapy from nutritional PN requirements.

Multiple sclerosis-related trigeminal neuralgia: a prospective series of 43 patients treated with gamma knife surgery with more than one year of follow-up.

BACKGROUND: Trigeminal neuralgia (TN) related to multiple sclerosis (MS) is more difficult to manage pharmacologically and surgically. OBJECTIVE: This article aims to evaluate the safety and efficacy of Gamma Knife surgery (GKS) in this special group of patients. METHODS: Between July 1992 and November 2010, 43 cases with more than 1 year of follow-up were operated with GKS for TN related to MS and prospectively evaluated in the Timone University Hospital, Marseille, France. Radiosurgery using the Gamma Knife (model B or C or Perfexion) was performed. A single 4-mm isocenter was positioned at a median distance of 8 mm (range 5.7-14.7) anterior to the emergence of the nerve. A median maximum dose of 85 Gy (range 75-90) was delivered. RESULTS: The median follow-up period was 53.8 months (12-157.1). Thirty-nine patients (90.7%) were initially pain free. Their actuarial probability of remaining pain free without medication at 6 months, 1, 3, 5 and 10 years was 87.2, 71.8, 43.1, 38.3 and 20.5%, respectively, and remained stable till 12 years. The hypoesthesia actuarial rate at 6 months, 1 and 2 years was 11.5, 11.5 and 16%, and remained stable till 12 years. CONCLUSIONS: GKS proved safe and effective in this special group of patients.

Previous microvascular decompression decreases the chances of pain free in patients treated with Gamma Knife radiosurgery for TIC

Object: The authors sought to establish whether the safety-efficacy of Gamma Knife radiosurgery (GKRS) as a second treatment for intractable trigeminal neuralgia (ITN) are influenced by prior microvascular decompression (MVD) which remains, for some of the authors, the reference technique. Methods: Between July 1992 and November 2010, 737 patients have been operated with GKRS for ITN and prospectively evaluated in Timone University Hospital in Marseille, France. Among these, 54 patients had a previous MVD history. Radiosurgery using a Gamma Knife (model B or C or Perfexion) was performed relying on both MR and CT targeting. A single 4 mm isocenter was positioned in the cisternal portion of the trigeminal nerve at a median distance of 7.6 mm (range 3.9- 11.9) anteriorly to the emergence of the nerve (retrogasserian target). A median maximum dose of 85 Gy (range 70-90) was delivered. Are further analyzed only 45 patients with previous MVD and a follow-up longer than one year (the patients with megadolichobasilar artery compression and multiple sclerosis were excluded). Results: The median age in this series was 56.75 years (range 28.09-82.39). The median follow-up period was 39.48 months (range 14.10-144.65). All the patients had a past history of surgery, with at least one previous failed MVD, but also a radiofrequency lesion (RFL) in 16 (35.6%) patients, balloon microcompression in 7 (15.6%) patients and glycerol rhizotomy in 1 case (2.2%). Thirty-five patients (77.8%) were initially pain free in a median time of 14 days (range 0, 180). Patients from this group had less probability of being pain free compared to our global population of essential trigeminal neuralgia without previous MVD history (p=0.010, hazard ratio of 0.64). Their probability of remaining pain free at 3, 5, 7 and 10 years was 66.5%, 59.1%, 59.1% and 44.3%, respectively. Twelve patients (34.3%) initially pain free experienced a recurrence with a median delay of 31.21 months (range 3.40-89.93). The hypoesthesia actuarial rate at 1 year was 9.1% and remained stable till 12 years with a median delay of onset of 8 months (range 8-8). Conclusions: Retrogasserian GKRS proofed to be safe and effective on the long-term basis even after failed previous MVD. Even if the initial result of pain free was of only 77.8%, the toxicity was low with only 9.1% hypoesthesia. No patient reported a bothersome hypoesthesia. The probability of maintaining pain relief in long-term was of 44.3% at 10 years.

A neuronal gamma oscillatory signature during morphological unification in the left occipitotemporal junction.

Morphology is the aspect of language concerned with the internal structure of words. In the past decades, a large body of masked priming (behavioral and neuroimaging) data has suggested that the visual word recognition system automatically decomposes any morphologically complex word into a stem and its constituent morphemes. Yet the reliance of morphology on other reading processes (e.g., orthography and semantics), as well as its underlying neuronal mechanisms are yet to be determined. In the current magnetoencephalography study, we addressed morphology from the perspective of the unification framework, that is, by applying the Hold/Release paradigm, morphological unification was simulated via the assembly of internal morphemic units into a whole word. Trials representing real words were divided into words with a transparent (true) or a nontransparent (pseudo) morphological relationship. Morphological unification of truly suffixed words was faster and more accurate and additionally enhanced induced oscillations in the narrow gamma band (60-85 Hz, 260-440 ms) in the left posterior occipitotemporal junction. This neural signature could not be explained by a mere automatic lexical processing (i.e., stem perception), but more likely it related to a semantic access step during the morphological unification process. By demonstrating the validity of unification at the morphological level, this study contributes to the vast empirical evidence on unification across other language processes. Furthermore, we point out that morphological unification relies on the retrieval of lexical semantic associations via induced gamma band oscillations in a cerebral hub region for visual word form processing.

Biomassa microbiana e produção de C-CO2 e N mineral de um podzólico vermelho-escuro submetido a diferentes sistemas de manejo: Microbial biomass and C-CO2 and mineral nitrogen production in paleudult soil cultivated under different management systems

Os sistemas de manejo, com diferenças no revolvimento do solo e na composição dos resíduos vegetais, alteram as propriedades biológicas do solo, com reflexos na qualidade do solo e na produtividade das culturas. Com vistas em medir estas alterações nas propriedades biológicas do solo, a biomassa e a atividade microbiana foram avaliadas em um Podzólico Vermelho-Escuro, em Eldorado do Sul (RS), utilizando diferentes preparos (convencional, reduzido e plantio direto) e dois sistemas de sucessões de culturas (aveia preta + vica/milho + caupi e aveia/milho). As avaliações foram realizadas em quatro épocas, durante 12 meses, e em duas profundidades (0-5 e 5-15 cm). O carbono da biomassa microbiana foi analisado pelo método de fumigação-incubação, e a atividade microbiana, pela produção de C-CO2 e N mineral, após 60 dias de incubação. As diferenças na biomassa e na atividade microbiana, entre os sistemas de manejo, foram mais pronunciadas na camada de 0-5 cm. Nesta camada de solo, observaram-se os maiores valores de biomassa e de atividade nos preparos conservacionistas e no sistema aveia + vica/milho + caupi. Dentre as variáveis estudadas, a mineralização de N mostrou-se a mais sensível aos manejos, à profundidade e à época de amostragem.

Mineralização do C e do N em amostras armazenadas de solo cultivado com cana-de-açúcar, ao longo de dez anos, com e sem fertilização nitrogenada

As taxas de mineralização do C e do N foram estimadas em amostras de um Podzólico Vermelho-Amarelo latossólico álico textura arenosa, retiradas nas profundidades de 0-20, 20-40 e 40-60 cm, ao longo dos 10 anos de um experimento de campo com cana-de-açúcar, de parcelas com e sem fertilização nitrogenada (60 kg ha-1 de N na forma de uréia). A mineralização do N foi medida por meio da técnica de incubação com percolação periódica e a do C pela quantidade de C-CO2 absorvida em solução de NaOH 1 mol L-1, por titulação potenciométrica. Em geral, apenas na camada superficial, o N total mineralizado acumulado durante as 20 semanas de incubação foi maior, 13% a mais no tratamento fertilizado que no não fertilizado. Entre épocas de amostragem, ao longo dos 10 anos, dentro de cada tratamento, houve diferenças significativas nas três profundidades. No entanto, as épocas de maiores mineralizações não foram as mesmas para todas as profundidades e tratamentos e não mostraram nenhuma tendência mais consistente. O C total mineralizado não diferiu significativamente entre os tratamentos (fertilizado e não fertilizado). As curvas de mineralização de C seguiram uma tendência mais linear que as do N, indicando uma possível estabilização nas taxas de mineralização entre 8 e 20 semanas. Os valores de produção média de C e N mineralizados foram de 611 e 26, 411 e 17 e 427 e 15 mg kg-1 de solo, para as profundidades de 0-20, 20-40 e 40-60 cm, respectivamente. O resultado mais importante foi a manutenção do potencial de mineralização de N do solo ao longo dos 10 anos de cultivo com cana, mesmo nas amostras provenientes das parcelas sem fertilização.

Cultivo consorciado de aveia e ervilhaca: relação C/N da fitomassa e produtividade do milho em sucessão

A aveia e a ervilhaca são as principais culturas de cobertura de solo utilizadas durante o inverno na região sul do Brasil. O cultivo consorciado dessas duas espécies, estratégia ainda pouco utilizada, pode potencialmente resultar numa fitomassa com relação C/N mais equilibrada do que aquela proveniente das culturas solteiras, bem como proporcionar resíduos culturais que atuem, simultaneamente, na proteção do solo contra os agentes erosivos e no suprimento de N ao milho. Com o objetivo de avaliar esta hipótese, realizou-se este trabalho, durante o ano agrícola de 1992/93, em área do Departamento de Solos da Universidade Federal de Santa Maria, num Argissolo Vermelho distrófico arênico (Hapludalf). Os tratamentos constaram de diferentes proporções de densidade de semeadura de ervilhaca comum (E) e aveia preta (A): T1: 100% E (80 kg ha-1de sementes); T2: 90% E (72 kg ha-1) + 10% A (8 kg ha-1); T3: 75% E (60 kg ha-1) + 25% A (20 kg ha-1); T4: 50% E (40 kg ha-1) + 50% A (40 kg ha-1); T5: 25% E (20 kg ha-1) + 75% A (60 kg ha-1) e T6: 100% A (80 kg ha-1). Além desses, foram utilizados dois tratamentos, nos quais o solo permaneceu em pousio durante o inverno: no primeiro tratamento, foram aplicados 75 kg ha-1 de N-uréia no milho (T7) e, no segundo, a cultura foi plantada sem adubação nitrogenada (T8). Em sucessão à ervilhaca como cultura solteira, com relação C/N de 13,5, a produtividade de grãos de milho chegou a 5,44 t ha-1, não diferindo do tratamento em pousio com aplicação de N-uréia. Todavia, a ervilhaca foi rapidamente decomposta e, seis meses após o seu manejo, apenas 19,5% da fitomassa inicial encontrava-se na superfície do solo. Com a inclusão da aveia em consórcio com a ervilhaca ocorreu um aumento gradativo na relação C/N da fitomassa, diminuindo o fornecimento de N ao milho e aumentando a persistência dos resíduos culturais. O consórcio que apresentou melhor equilíbrio entre produção de fitomassa, proteção do solo pelos resíduos culturais e fornecimento de N ao milho foi o que continha 10% de aveia + 90% de ervilhaca (relação C/N = 18,6).

The synthetic Plasmodium falciparum circumsporozoite peptide PfCS102 as a malaria vaccine candidate: a randomized controlled phase I trial.

BACKGROUND: Fully efficient vaccines against malaria pre-erythrocytic stage are still lacking. The objective of this dose/adjuvant-finding study was to evaluate the safety, reactogenicity and immunogenicity of a vaccine candidate based on a peptide spanning the C-terminal region of Plasmodium falciparum circumsporozoite protein (PfCS102) in malaria naive adults. METHODOLOGY AND PRINCIPAL FINDINGS: Thirty-six healthy malaria-naive adults were randomly distributed into three dose blocks (10, 30 and 100 microg) and vaccinated with PfCS102 in combination with either Montanide ISA 720 or GSK proprietary Adjuvant System AS02A at days 0, 60, and 180. Primary end-point (safety and reactogenicity) was based on the frequency of adverse events (AE) and of abnormal biological safety tests; secondary-end point (immunogenicity) on P. falciparum specific cell-mediated immunity and antibody response before and after immunization. The two adjuvant formulations were well tolerated and their safety profile was good. Most AEs were local and, when systemic, involved mainly fatigue and headache. Half the volunteers in AS02A groups experienced severe AEs (mainly erythema). After the third injection, 34 of 35 volunteers developed anti-PfCS102 and anti-sporozoite antibodies, and 28 of 35 demonstrated T-cell proliferative responses and IFN-gamma production. Five of 22 HLA-A2 and HLA-A3 volunteers displayed PfCS102 specific IFN-gamma secreting CD8(+) T cell responses. Responses were only marginally boosted after the 3(rd) vaccination and remained stable for 6 months. For both adjuvants, the dose of 10 microg was less immunogenic in comparison to 30 and 100 microg that induced similar responses. AS02A formulations with 30 microg or 100 microg PfCS102 induced about 10-folds higher antibody and IFN-gamma responses than Montanide formulations. CONCLUSIONS/SIGNIFICANCE: PfCS102 peptide was safe and highly immunogenic, allowing the design of more advanced trials to test its potential for protection. Two or three immunizations with a dose of 30 microg formulated with AS02A appeared the most appropriate choice for such studies. TRIAL REGISTRATION: Swissmedic.ch 2002 DR 1227.