International Tailored Chemotherapy Adjuvant (ITACA) trial, a phase III multicenter randomized trial comparing adjuvant pharmacogenomic-driven chemotherapy versus standard adjuvant chemotherapy in completely resected stage II-IIIA non-small-cell lung cancer.

BACKGROUND: Several strategies have been investigated to improve the 4% survival advantage of adjuvant chemotherapy in early-stage non-small-cell lung cancer (NSCLC). In this investigator-initiated study we aimed to evaluate the predictive utility of the messenger RNA (mRNA) expression levels of excision repair cross complementation group 1 (ERCC1) and thymidylate synthase (TS) as assessed in resected tumor. PATIENTS AND METHODS: Seven hundred and seventy-three completely resected stage II-III NSCLC patients were enrolled and randomly assigned in each of the four genomic subgroups to investigator's choice of platinum-based chemotherapy (C, n = 389) or tailored chemotherapy (T, n = 384). All anticancer drugs were administered according to standard doses and schedules. Stratification factors included stage and smoking status. The primary endpoint of the study was overall survival (OS). RESULTS: Six hundred and ninety patients were included in the primary analysis. At a median follow-up of 45.9 months, 85 (24.6%) and 70 (20.3%) patients died in arms C and T, respectively. Five-year survival for patients in arms C and T was of 65.4% (95% CI (confidence interval): 58.5% to 71.4%) and 72.9% (95% CI: 66.5% to 78.3%), respectively. The estimated hazard ratio (HR) was 0.77 (95% CI: 0.56-1.06, P value: 0.109) for arm T versus arm C. HR for recurrence-free survival was 0.89 (95% CI: 0.69-1.14, P value: 0.341) for arm T versus arm C. Grade 3-5 toxicities were more frequently reported in arm C than in arm T. CONCLUSION: In completely resected stage II-III NSCLC tailoring adjuvant chemotherapy conferred a non-statistically significant trend for OS favoring the T arm. In terms of safety, the T arm was associated with better efficacy/toxicity ratio related to the different therapeutic choices in the experimental arm.

Semiquantitative analysis of interim 18F-FDG PET is superior in predicting outcome in Hodgkin lymphoma patients compared to visual analysis.

PURPOSE: To investigate the prognostic value of interim PET (PETi) in adult HL patients, comparing visual with semiquantitative analysis. MATERIAL AND METHODS: Retrospective analysis of Hodgkin's lymphoma (HL) patients diagnosed between 2012 and 2016 in the Onco-hematology Department of Instituto Português de Oncologia - Porto (median follow-up: 46.5 months [2.6-66.4]). Fifty-eight patients with available PET at diagnosis (PET0) and PETi data were included. PETi scans were analyzed according to Deauville 5-point scale (5-PS), and cut-off values for changes in maximum standardized uptake value [SUVmax], peak SUV [SUVpeak], metabolic tumour volume [MTV] and total lesion glycolysis index [TLG] between PETi and PET0 were computed using ROC analysis. Visual and semiquantitative data were compared with each other in the prediction of patient outcomes. RESULTS: Semiquantitative analysis obtained a higher sensitivity for persistent/relapsed disease compared to the 5-PS (70% vs. 10%, respectively), but lower specificity. It also demonstrated better predictive performance for response to first-line therapy (negative predictive value >92%). The positive predictive value was similar for all five measurements. At 60 months of follow-up, there was a significant difference between the progression free survival (PFS) curves of patients with positive and negative PETi according to ΔSUVmax (56.9% vs. 88.0%, p<0.05), ΔSUVpeak (55.9% vs. 88.1%, p<0.05), ΔMTV (35.3% vs. 88.7%, p<0.05), and ΔTLG (42.4% vs. 88.1%, p<0.05). Statistical significance was not reached when considering 5-PS results. DISCUSSION: PETi interpretation according to a semiquantitative approach appears to discriminate HL patients better than the visual 5-PS analysis. This could allow better detection of persistent or early relapsed disease, while a negative PETi result could support de-escalating therapy intensity.

Association between Bone Mineral Density and Nutritional Status, Body Composition and Bone Metabolism in Older Adults.

PURPOSE: To identify an association between bone mineral density (BMD) and nutritional status, body composition and bone metabolism in older patients. METHODS: Cross-sectional study, involving older adults, with osteopenia/osteoporosis and with normal BMD. The mineral density of the lumbar spine from L1 to L4 and the proximal region of the femur was assessed using dual energy X-ray absorptiometry. Biochemical analyzes were performed of 25(OH)-D, calcium and parathormone. Weight, knee height, and abdominal (AC), mid-upper arm (MUAC) and calf (CC) circumferences were measured. The percentage of body fat (%BF) and Fat-Free Mass (FFM) were quantified by electrical bioimpedance analysis. The Body Mass Index (BMI) was calculated. The statistical analysis used bivariate and multivariate, parametric and/or non-parametric tests, and was considered significant when p <0.05. RESULTS: Of the total 51 older adults assessed, 30 of them (58.8%) were diagnosed with osteopenia/osteoporosis. Body weight (p = 0.001), BMI (p = 0.001), % BF (p = 0.030) and serum concentrations of 25(OH)-D (p = 0.003) were higher in the group without changes in BMD. BMI and serum levels of 25(OH)-D demonstrated a positive correlation with the BMD of all bone compartments and the AC displayed a positive correlation with the lumbar vertebrae. In the logistic regression models, adjusted for sex and age, the BMI and the serum concentration of 25(OH)-D were presented as a protective factor against osteopenia/osteoporosis. CONCLUSIONS: Higher body weight, BMI, AC and %BF, and sufficient serum levels of vitamin D, were shown to be promoters of BMD.

Mapping of Texture and Phase Fractions in Heterogeneous Stress States during Multiaxial Loading of Biomedical Superelastic NiTi.

Thermoelastic deformation mechanisms in polycrystalline biomedical-grade superelastic NiTi are spatially mapped using in situ neutron diffraction during multiaxial loading and heating. The trigonal R-phase is formed from the cubic phase during cooling to room temperature and subsequently deforms in compression, tension, and torsion. The resulting R-phase variant microstructure from the variant reorientation and detwinning processes are equivalent for the corresponding strain in tension and compression, and the variant microstructure is reversible by isothermal loading. The R-phase variant microstructure is consistent between uniaxial and torsional loading when the principal stress directions of the stress state are considered (for the crystallographic directions observed here). The variant microstructure evolution is tracked and the similarity in general behavior between uniaxial and torsional loading, in spite of the implicit heterogeneous stress state associated with torsional loading, pointed to the ability of the reversible thermoelastic transformation in NiTi to accommodate stress and strain mismatch with deformation. This ability of the R-phase, despite its limited variants, to accommodate stress and strain and satisfy strain incompatibility in addition to the existing internal stresses has significance for reducing irrecoverable deformation mechanisms during loading and cycling through the phase transformation.

Comparison between infrared and hot-blade beak trimming methods in white and brown pullets: performance, organ and bone development / [Comparativo entre debicagem infravermelha e por lâmina-quente em linhagens de frangas leves e semi-pesadas: desempenho, desenvolvimento ósseo e de órgãos]

The purpose of this study was to evaluate infrared beak trimming versus hot-blade beak trimming on performance, organ and bone development of laying pullets, using 240 birds, at 0-16 weeks of age, from two strains (white layer - Hy-Line W36 and brown layer - Hy-Line Brown). Infrared debeaking was performed on the first day of life in the hatchery, whereas hot-blade beak trimming was performed at eight days of age. Zootechnical performance at 6, 9, 12 and 15 weeks of age was analyzed. At the end of the trial, one Bird per experimental unit was euthanized for liver, gizzard, proventriculus, heart and small intestine relative weight evaluation. Both tibiotarsus were collected to measure bone breaking resistance, bone mineral content and bone densitometry. There was higher body weight and feed intake in both bird strains submitted to infrared beak trimming. Additionally, hot-blade white layers presented lower body development and feed intake. The debeaking methods did not influence relative organ weights or tibiotarsus parameters evaluated. We conclude that the infrared beak trimming method promotes better pullet body development than the conventional hot-blade method, improving feed intake, without compromising organ and bone development, being a more adequate debeaking method to be used in layer production.(AU)

Para avaliar a debicagem infravermelha versus a debicagem pelo método da lâmina quente no desempenho, no desenvolvimento de órgãos e na qualidade óssea de poedeiras em recria, foram utilizadas 240 aves, entre zero e 16 semanas de idade, em duas linhagens (branca - Hy-Line W36 e vermelha - Hy-Line Brown). A debicagem por infravermelho foi realizada no primeiro dia de vida no incubatório, enquanto a debicagem com lâmina quente foi realizada aos oito dias de idade. Analisou-se o desempenho zootécnico na sexta, nona, 12ª e 15ª semanas de idade. No final do estudo, uma ave por unidade experimental foi eutanasiada para avaliação do peso relativo do fígado, da moela, do proventrículo, do coração e do intestino delgado, e ossos do tibiotarso foram coletados para avaliação da resistência à ruptura, do conteúdo mineral e da densitometria óssea. Houve maior peso corporal e consumo alimentar nas aves pertencentes às duas linhagens quando submetidas à debicagem por infravermelho, sem haver influência sobre o peso relativo dos órgãos e a qualidade óssea. Conclui-se que o método de debicagem por radiação infravermelha promove melhor desenvolvimento corporal das frangas em relação à debicagem por lâmina quente, melhorando consumo de ração, sem comprometer o desenvolvimento de órgãos e ossos, sendo mais adequada para utilização na avicultura de postura comercial.(AU)

Metronomic oral vinorelbine for the treatment of advanced non-small cell lung cancer: a multicenter international retrospective analysis.

PURPOSE: Metronomic oral vinorelbine (MOV) could be a treatment option for unfit patients with advanced non-small cell lung cancer (NSCLC) based on its safety profile and high patient compliance. METHODS: We retrospectively collected data on 270 patients [median age 76 (range 48-92) years, M/F 204/66, PS 0 (27)/1 (110)/≥ 2 (133), median of 3 serious comorbidities] with stage IIIB-IV NSCLC treated with MOV as first (T1) (67%), second (T2) (19%) or subsequent (T3) (14%) line. Schedules consisted of vinorelbine 50 mg (138), 40 mg (68) or 30 mg (64) three times a week continuously. RESULTS: Patients received an overall median of 6 (range 1-25) cycles with a total of 1253 cycles delivered. The overall response rate was 17.8% with 46 partial and 2 complete responses and 119 patients (44.1%) experienced stable disease > 12 weeks with an overall disease control rate of 61.9%. Median overall time to progression was 5 (range 1-21) months [T1 7 (1-21), T2 5.5 (1-19) and T3 4 (1-19) months] and median overall survival 9 (range 1-36) months [T1 10 (1-31), T2 8 (1-36) and T3 6.5 (2-29) months]. Treatment was extremely well tolerated with 2% (25/1253) G3/4 toxicity (mainly G3 fatigue and anemia) and no toxic deaths. We observed the longer OS 14 (range 7-36) months in a subset of squamous NSCLC patients receiving immunotherapy after metronomic oral vinorelbine. CONCLUSION: We confirmed MOV as an extremely safe treatment in a large real world population of advanced NSCLC with an interesting activity mainly consisting of long-term disease stabilization. We speculate the possibility of a synergistic effect with subsequent immunotherapy.

Inter- and intra-variability of seed germination traits of Carpobrotus edulis N.E.Br. and its hybrid C. affine acinaciformis.

Invasions by alien Carpobrotus spp. have been recognised as one of the most severe threats to Mediterranean climate coastal ecosystems, and Carpobrotus is considered one of the most widespread invasive alien genera in the Mediterranean Basin. The aims of this study were to characterise seed germination of both C. edulis and its hybrid C. aff. acinaciformis, in terms of photoperiod, temperature and salinity. Inter- and intra-specific variability in the responses to photoperiod (12/12 h light and total darkness), constant temperatures (5, 10, 15, 20, 25 °C) and an alternating temperature regime (25/10 °C), salt stress (0, 125, 250, 500 mm NaCl) and the recovery of seed germination were evaluated for two seed lots of C. edulis and two of its hybrid C. aff. acinaciformis. All the tested seed lots achieved higher germination percentages in the light, with respect to total darkness. In relation to temperature, the two C. edulis seed lots did not show a preference, while the two C. aff. acinaciformis seed lots differed in their germination response, one germinating more at the lowest temperatures (5 and 10 °C) and one at the highest (20 and 25 °C). For all seed lots, highest germination occurred without NaCl (0 mm) and germination decreased with increasing salinity. Different germination requirements in a saline substrate were not detected for C. edulis, while they were observed for C. aff. acinaciformis. Marked differences were detected in recovery responses between the two taxa. C. edulis has the ability to germinate over a wide time window throughout the year. This study identified significant differences in seed production, seed mass, germination requirements (temperature) and salinity tolerance for both C. edulis and C. aff. acinaciformis. Our results indicate the extreme versatility of the hybrid forms to germinate in a wide range of natural conditions and habitats.

Pain-motor integration in the primary motor cortex in Parkinson's disease.

BACKGROUND: In Parkinson's disease (PD), the influence of chronic pain on motor features has never been investigated. We have recently designed a technique that combines nociceptive system activation by laser stimuli and primary motor cortex (M1) activation through transcranial magnetic stimulation (TMS), in a laser-paired associative stimulation design (Laser-PAS). In controls, Laser-PAS induces long-term changes in motor evoked potentials reflecting M1 long-term potentiation-like plasticity, arising from pain-motor integration. OBJECTIVE: We here examined the possible influence of chronic pain on motor responses to Laser-PAS in patients with PD, with and without chronic pain. METHODS: We compared motor responses to Laser-PAS in healthy subjects and in patients with PD, with and without chronic pain. RESULTS: Unlike controls, we found reduced responses to Laser-PAS in patients with PD, with and without pain. Patients off and on dopaminergic therapy had similar responses to Laser-PAS. When comparing responses to Laser-PAS in patients with and without pain, the two patients' subgroups had similar abnormalities. When we compared patients with pain in the body region investigated with Laser-PAS, with those with pain in other body regions, we found prominent changes in patients with homotopic pain. Finally, when comparing Laser-PAS with the original PAS protocol, which combines electric peripheral nerve stimuli and TMS, in patients without pain and those with homotopic pain, we found similar responses to both techniques in patients without pain, whereas Laser-PAS induced greater abnormalities than PAS in patients with pain. CONCLUSIONS: In PD, chronic pain degrades response to Laser-PAS through abnormal pain-motor integration.

Fracionamento de carboidratos e proteínas, cinética de degradação ruminal in vitro pela técnica de produção de gás, de rações suplementares contendo torta de girassol / Fractionation of carbohydrates, protein and "in vitro" ruminal degradation kinetics by gas production technique of supplementary rations containing sunflower cake

Objetivou-se neste estudo fracionar os carboidratos e as proteínas e avaliar a cinética de degradação ruminal de rações contendo 0; 24; 48 e 72% de torta de girassol. Os carboidratos foram fracionados em A+B1, B2 e C, e as proteínas em A, B1+B2, B3 e C. Os parâmetros de cinética de degradação ruminal das rações foram estimados a partir do método semiautomático de produção cumulativa de gases in vitro. Foram utilizados quatro animais 1/2 Gir x 1/2 Holandês, machos e castrados, fistulados no rúmen, mantidos em pastagem e recebendo 1kg de ração concentrada, como doadores de inócuo ruminal. O ensaio foi realizado em delineamento em blocos ao acaso, com quatro rações e quatro períodos, e os resultados obtidos foram interpretados de acordo com a análise de variância e de regressão a 5% de significância. Houve efeito linear decrescente para o fracionamento de carboidratos totais e a fração A+B1, e efeito linear crescente sobre as frações B2 e C. Para as frações nitrogenadas, houve efeito linear crescente sobre a fração A, e efeito linear decrescente sobre as frações B1+B2 e C. Em relação à cinética de degradação ruminal in vitro, houve efeito decrescente sobre o parâmetro "a", e efeito quadrático sobre os parâmetros "b" e "c", com pontos de máxima, respectivamente, de 61,84 e 66,77% de torta de girassol. A inclusão da torta de girassol nas rações suplementares, nos teores estudados, influenciou nas frações dos carboidratos, das proteínas e nos parâmetros de degradação ruminal. Caso haja a disponibilidade desse ingrediente, ele pode ser adicionado à ração suplementar de bovinos sem prejuízo nos parâmetros nutricionais e metabólicos.(AU)

This work aimed to fractionate carbohydrates and protein and to evaluate ruminal degradation kinetics of rations containing 0; 24; 48 and 72% of inclusion of sunflower cake. The carbohydrates were fractionate in A+B1, B2 and C fraction, and protein in A, B1+B2, B3 and C. Rations ruminal degradation kinetics were realized with the "in vitro" semiautomatic gas production technique. Four male, castrated, 1/2 Gir x 1/2 Holstein were used, all of them fistulated in the rumen. This study was done in randomized block design with four rations and four periods. The results obtained were interpreted according to variance and regression analysis at 5% significance level. There is a decreasing effect for total carbohydrates fractionation and A+B1 fraction, and there is an increasing effect for B2 and C fraction. For nitrogen fractions there is an increasing effect on fraction A and negative linear effect on B1+B2 and C fractions. For "In vitro" ruminal degradation kinetics there is a decreasing effect for "a" parameter, and quadratic effect for "b" and "c" parameters with maximum points respectively 61.84 and 66.77% of sunflower seed meal. The inclusion of sunflower cake in supplementary feeding, the studied levels, influenced the fractions of carbohydrates, proteins and degradation parameters. If there is the availability of this ingredient, it can be added to the supplemental feed cattle subject to the nutritional and metabolic parameters.(AU)

Effects of NaCl stress on seed germination and seedling development of Brassica insularis Moris (Brassicaceae).

Brassica insularis is a protected plant that grows on both coastal and inland cliffs in the western Mediterranean Basin. The objective of this study was to test if any variability exists in the salt stress response during seed germination and seedling development in this species relative to its provenance habitat. Variability among three populations in the salt stress effects on seed germination and recovery under different temperatures was evaluated. The effect of nebulisation of a salt solution on seedling development was evaluated between populations growing at different distances from the sea. Seeds of B. insularis could germinate at NaCl concentrations up to 200 mm. Seed viability was negatively affected by salt, and recovery ability decreased with increasing temperature or salinity. Inter-population variability was detected in salt response during the seed germination phase, as well as in seedling salt spray tolerance. The inland population seedlings had drastically decreased survival and life span and failed to survive to the end of the experiment. In contrast, at least 90% of the coastal seedlings survived, even when sprayed at the highest frequency with salt solution. This study allowed investigation of two natural factors, soil salinity and marine aerosols, widely present in the B. insularis habitat, and provided the first insights into ecology of this protected species and its distribution in the Mediterranean. These results might be useful in understanding the actual distributions of other species with the same ecology that experience these same abiotic parameters.

ALK gene copy number gains in non-small-cell lung cancer: prognostic impact and clinico-pathological correlations.

BACKGROUND: The correlation between ALK gene copy number gain (ALK-CNG) and prognosis in the context of advanced non-small-cell lung cancer (NSCLC) remains a controversial issue. This study aimed to evaluate the association among ALK-CNG according to Fluorescent In Situ Hybridization (FISH), clinical characteristics and survival in resectable and advanced NSCLC. METHODS: Clinical and pathological data of patients with resectable and advanced NSCLC were retrospectively collected. Tumor tissues were analyzed for ALK-CNG by FISH, and patients were divided in 3 groups/patterns on the basis of ALK signals: disomic [Pattern A], 3-7 signals [Pattern B], >7 signals [Pattern C]. The association between clinical and pathological features and ALK-CNG patterns was evaluated. Disease/progression-free and overall survival (DFS/PFS and OS) were estimated using the Kaplan-Meyer method. RESULTS: A number of 128 (76.6 %) out of the 167 eligible patients were evaluable for ALK-CNG, displaying pattern A, B and C in 71 (42.5 %), 42 (25.1 %) and 15 (9 %) patients, respectively. Gains in ALK-CNG appear to be more frequent in smokers/former smokers than in non-smokers (74.2 % versus 20.4 %, respectively, p = 0.03). Pattern A and C seem more frequently associated with higher T-stage (T3-4), while pattern B appears more represented in lower T-stage (T 1-2) (p = 0.06). No significant differences in survival rate were observed among the above groups. CONCLUSIONS: A high ALK-CNG pattern might be associated with smoking status and theoretically it might mirror genomic instability. The implications for prognosis should be prospectively investigated and validated in larger patients' series. TRIAL REGISTRATION: We confirm that all the study was performed in accordance with relevant guidelines and regulations and that all the protocol (part of a larger project MFAG 2013 N.14282) was approved by the local Ethics Committee of the Azienda Ospedaliera Universitaria Integrata of Verona on November 11st, 2014.

Roflumilast inhibits leukocyte-platelet interactions and prevents the prothrombotic functions of polymorphonuclear leukocytes and monocytes.

UNLABELLED: ESSENTIALS: Thrombosis is a major comorbidity in patients with chronic obstructive pulmonary disease (COPD). Roflumilast is a selective phosphodiesterase type-4 (PDE4) inhibitor approved for treatment of severe COPD. PDE4 blockade by roflumilast inhibits prothrombotic functions of neutrophils and monocytes. PDE4 inhibitors may reduce thrombotic risk in COPD as well as in other vascular diseases. BACKGROUND: Roflumilast, an oral selective phosphodiesterase type 4 inhibitor, is approved for the treatment of severe chronic obstructive pulmonary disease (COPD). A recent meta-analysis of trials on COPD revealed that treatment with roflumilast was associated with a significant reduction in the rate of major cardiovascular events. The mechanisms of this effect remain unknown. OBJECTIVES: We tested the hypothesis that roflumilast N-oxide (RNO), the active metabolite of roflumilast, curbs the molecular mechanisms required for leukocyte-platelet (PLT) interactions and prevents the prothrombotic functions of polymorphonuclear leukocytes (PMNs) and monocytes (MNs). METHODS: Using well-characterized in vitro models, we analysed the effects of RNO on: (i) PMN adhesiveness; (ii) the release of neutrophil extracellular traps (NETs); and (iii) tissue factor expression in MNs. Key biochemical events underlying the inhibitory effects of RNO were defined. RESULTS AND CONCLUSIONS: In PMNs, RNO prevented phosphoinositide 3-kinase (PI3K)-dependent phosphorylation of Akt on Ser473, and Src family kinase (SFK)-mediated Pyk2 phosphorylation on Tyr579-580, while inducing protein kinase A-mediated phosphorylation of C-terminal Src kinase, the major negative regulator of SFKs. Modulation of these signaling pathways by RNO resulted in a significant impairment of PMN adhesion to activated PLTs or human umbilical vein endothelial cells, mainly mediated by inhibition of the adhesive function of Mac-1. Moreover RNO curbed SFK/PI3K-mediated NET release by PMNs adherent on fibrinogen-coated surfaces. In MNs interacting with activated PLTs, RNO curbed PI3K-mediated expression of tissue factor. The efficacy of RNO was significantly potentiated by formoterol, a long acting ß-adrenergic receptor agonist. This study reveals novel antithrombotic activities by which roflumilast may exert protective effects against cardiovascular comorbodities in COPD.

Selective inhibition of TNFR1 reduces osteoclast numbers and is differentiated from anti-TNF in a LPS-driven model of inflammatory bone loss.

The treatment of autoimmune disorders has been revolutionised by the introduction of biologics such as anti-tumour necrosis factor (anti-TNF). Although in rheumatoid arthritis patients a bone sparing effect of anti-TNF has been shown, the mechanism is not fully understood. Anti-TNF molecules block tumour necrosis factor (TNF) and prevent signalling via both TNF receptor 1 (TNFR1; p55) and TNF receptor 2 (TNFR2; p75). However, signalling via TNFR2 is reported to have protective effects in a number of cell and organ systems. Hence we set out to investigate if pharmacological inhibition of TNFR1 had differential effects compared to pan-TNF inhibition in both an in vitro cell-based model of human osteoclast activity and an in vivo mouse model of lipopolysaccharide (LPS)-induced osteolysis. For the in vitro experiments the anti-human TNFR1 domain antibody (dAb) DMS5541 was used, whereas for the in vivo mouse experiments the anti-mouse TNFR1 dAb DMS5540 was used. We show that selective blocking of TNFR1 signalling reduced osteoclast formation in the presence of TNF. Subcutaneous LPS injection over the calvaria leads to the development of osteolytic lesions within days due to inflammation driven osteoclast formation. In this model, murine TNFR2 genetically fused with mouse IgG1 Fc domain (mTNFR2.Fc), an anti-TNF, did not protect from bone loss in contrast to anti-TNFR1, which significantly reduced lesion development, inflammatory infiltrate, and osteoclast number and size. These results support further exploring the use of TNFR1-selective inhibition in inflammatory bone loss disorders such as osteomyelitis and peri-prosthetic aseptic loosening.

Morpho-colorimetric analysis and seed germination of Brassica insularis Moris (Brassicaceae) populations.

Brassica insularis is a perennial plant growing on both coastal and inland cliffs. Three seed lots from Sardinia were analysed using an image analysis system to detect differences in seed morphology, both within and among populations. Germination requirements at constant (5-25 °C) and alternating temperatures (25/10 °C), both in light and in darkness, were evaluated for all populations. In addition, the effect of a dry after-ripening period (90 days at 25 °C) was also investigated. Morpho-colorimetric analysis clearly identified seeds from different populations and discriminated three chromatic categories for seeds belonging to the Isola dei Cavoli coastal population, but not for the inland Masùa and the coastal Planu Sartu. Inter-population variability was also observed in germination behaviour. B. insularis seeds germinated, with percentages up to 60%, in a wide range of temperatures (5-25 °C), and neither light nor dry after-ripening affected final germination percentages. Moisture content measurements were made for seeds of each colour, but there were no particular differences among colours. Inter-populational variability in germination behaviour may be a survival strategy for species growing under unpredictable environmental conditions, such as under Mediterranean climate, while heteromorphy may be due to independent evolutionary divergence processes of the Isola dei Cavoli population.

Carboplatin and gemcitabine in first-line treatment of elderly patients with advanced non-small cell lung cancer: data from a retrospective study.

We retrospectively evaluated elderly patients with advanced non-small cell lung cancer (NSCLC) treated with carboplatin (AUC 4-5) and gemcitabine (1,000-1,200 mg/m²). Thirty-six patients with performance Status (pS) 0-1 and median age 73 (range 70-78 years) were considered. Histology was squamous cell carcinoma (8 patients), adenocarcinoma (22) and NSCLC not otherwise specified (6). 149 cycles of chemotherapy were administered with a median of 3 per patient (range 3-6). Grade 3 non-hematologic toxicities were dyspnea (1 patient) and fever (1). Grade 3/4 hematologic toxicities were anemia (6), neutropenia (6) and thrombocytopenia (10), with dose reduction required in 13 patients. The overall disease control rate was 44.4%. We recorded no complete response, 8 partial response, 8 stable disease and 20 progressive disease. After a medium follow-up of 11 months, median progression- free survival and median survival were 5 and 11 months, respectively. Carboplatin and gemcitabine is a safe and active regimen in elderly advanced NSCLC patients with good PS.

Glycogen synthase kinase-3 negatively regulates tissue factor expression in monocytes interacting with activated platelets.

BACKGROUND: At the site of vascular injury, monocytes (MN) interacting with activated platelets (PLT) synthesize tissue factor (TF) and promote thrombus formation. Intracellular signals necessary for the expression of TF in MN, in the context of a developing thrombus, remain unknown. OBJECTIVE: The study was designed to investigate the role of the glycogen synthase kinase 3 (GSK3, a serine-threonine kinase) downstream insulin receptor pathway, in PLT-induced TF expression in MN. METHODS: To this purpose we used a well-characterized in vitro model of human MN-PLT interactions that allows detailed analysis of TF activity, TF protein and gene expression. RESULTS: The results demonstrated that, in MN interacting with activated PLT: (i) TF activity, antigen and mRNA were low until 8-10 h and dramatically increased thereafter, up to 24 h; (ii) according to the kinetics of TF expression in MN, GSK3ß undergoes phosphorylation on serine 9, a process associated with down-regulation of enzyme activity; (iii) pharmacological blockade of GSK3 further increased TF expression and was accompanied by increased accumulation of NF-kB, in the nucleus; (iv) blockade of phosphoinositide-3 kinase (PI(3)K) by wortmannin inhibited PLT-induced TF expression; and (v) according to the established role of the GSK3 downstream insulin receptor, insulin increased PLT-induced TF expression in a PI(3)K-dependent manner. CONCLUSION: GSK3 acts as a molecular brake on the signaling pathway, leading to TF expression in MN interacting with activated PLT. PI(3)K, through Akt-dependent phosphorylation of GSK3, relieves this brake and allows TF gene expression. This study identifies a novel molecular link between thrombotic risk and metabolic disorders.

A study in situ of the effect of metallo- and serine proteinase inhibitors on the birefringence of the secretory stage enamel organic extracellular matrix.

Dental enamel formation occurs extracellularly and establishment of an ordered enamel organic extracellular matrix (ECM) seems to be crucial for proper construction of the enamel mineral phase. Polarizing microscopy shows that the ordered supramolecular structure of the secretory stage enamel organic ECM exhibits strong birefringence. We reported earlier that this birefringence is lost in unfixed specimens, probably due to extensive proteolytic cleavage of enamel proteins. Therefore, we investigated the association between enamel proteinase activities by analyzing the effects of metallo- and serine proteinase inhibitors in situ on the birefringence of the secretory stage enamel organic ECM. Male rats were used in the present study. After sacrifice, distal 10 mm fragments of upper incisors were removed and immersed for 15 h under continuous shaking at 37°C in one of the following solutions: 1) 10 mM Tris, pH 8.0; 150 mM NaCl (negative control, n = 8); 2) 2% paraformaldehyde and 0.5% glutaraldehyde in 0.2 M phosphate-buffered saline (PBS), pH 7.2 (positive control, n = 5); 3) 10 mM Tris, pH 8.0; 150 mM NaCl; 2 mM 1,10-phenanthroline (n = 9); 4) 10 mM Tris, pH 8.0; 150 mM NaCl; 2 mM phenylmethyl-sulfonyl fluoride (PMSF) (n = 8); 5) 10 mM Tris, pH 8.0; 150 mM NaCl; 2 mM 1,10-phenanthroline; 2 mM PMSF (n = 9). Samples then were immersed in fixative solution for 24 h and processed to obtain 5 µm thick longitudinal sections of the secretory stage enamel organic ECM. The sections were immersed in 80% glycerin for 30 min and analyzed by transmitted polarizing light microscopy. 1,10-Phenanthroline (inhibitor of metalloproteinases) and 1,10-phenanthroline + PMSF (inhibitor of serine proteinases) clearly prevented a decrease in the optical retardation of birefringence brightness from the tissue. PMSF alone promoted a slight preservation of the birefringence exhibited by the secretory stage enamel organic ECM. Rapid loss of birefringence in secretory stage enamel organic ECM that is not fixed immediately is caused by enamel proteinases and the activity of metalloproteinases seems to lead to preliminary degradation of the enamel organic ECM, which in turn facilitates subsequent serine proteinase activity.

Trends in systemic lupus erythematosus mortality rates in the state of Sao Paulo, Brazil from 1985 to 2004.

OBJECTIVES: To estimate mortality rates and mortality trends from SLE in the state of São Paulo, Brazil. MATERIAL AND METHODS: The official data bank was used to study all deaths occurred from 1985 to 2004 in which SLE was mentioned as the underlying cause of death. Besides the overall mortality rate, the annual gender- and age-specific mortality rates were estimated for each calendar year by age bracket (0-19 years, 20-39 years, 40-59 years and over 60 years) and for the sub-periods 1985-1995 (first) and 1996-2004 (second), by decades. Chi-square test was used to compare the mortality rates between the two periods, as well the mortality rates according to educational level considering years of study. Pearson correlation coefficient test was used to analyse mortality trends. The crude rates were adjusted for age by the direct method, using the standard Brazilian population in 2000. RESULTS: A total of 2,601 deaths (90% female) attributed to SLE were analysed. The mean age at death was significantly higher in the second than in the first sub-period (36.6+/-15.6 years vs. 33.9+/-14.0 years; p<0.001). The overall adjusted mortality rate was 3.8 deaths/million habitants/year for the entire period and 3.4 deaths/million inhabitants/year for the first and 4.0 deaths/million inhabitants/year for the second sub-period (p<0.001). In each calendar year, the mortality rate was significantly lower for the better educated group. Throughout the period, there was a significant increase in mortality rates only among women over 40. CONCLUSIONS: SLE patients living in the state of São Paulo still die at younger ages than those living in developed countries. Our data do not support the theory that there was an improvement in the SLE mortality rate in the last 20 years in the state of Sao Paulo. Socio-economic factors, such as the difficulty to get medical care and adequate treatment, may be the main factors to explain the worst prognosis for our patients.