Updated overall survival and final progression-free survival data for patients with treatment-naive advanced ALK-positive non-small-cell lung cancer in the ALEX study.

BACKGROUND: The ALEX study demonstrated significantly improved progression-free survival (PFS) with alectinib versus crizotinib in treatment-naive ALK-positive non-small-cell lung cancer (NSCLC) at the primary data cut-off (9 February 2017). We report mature PFS (cut-off: 30 November 2018) and overall survival (OS) data up to 5 years (cut-off: 29 November 2019). PATIENTS AND METHODS: Patients with stage III/IV ALK-positive NSCLC were randomized to receive twice-daily alectinib 600 mg (n = 152) or crizotinib 250 mg (n = 151) until disease progression, toxicity, withdrawal or death. Primary end point: investigator-assessed PFS. Secondary end points included objective response rate, OS and safety. RESULTS: Mature PFS data showed significantly prolonged investigator-assessed PFS with alectinib [hazard ratio (HR) 0.43, 95% confidence interval (CI) 0.32-0.58; median PFS 34.8 versus 10.9 months crizotinib]. Median duration of OS follow-up: 48.2 months alectinib, 23.3 months crizotinib. OS data remain immature (37% of events). Median OS was not reached with alectinib versus 57.4 months with crizotinib (stratified HR 0.67, 95% CI 0.46-0.98). The 5-year OS rate was 62.5% (95% CI 54.3-70.8) with alectinib and 45.5% (95% CI 33.6-57.4) with crizotinib, with 34.9% and 8.6% of patients still on study treatment, respectively. The OS benefit of alectinib was seen in patients with central nervous system metastases at baseline [HR 0.58 (95% CI 0.34-1.00)] and those without [HR 0.76 (95% CI 0.45-1.26)]. Median treatment duration was longer with alectinib (28.1 versus 10.8 months), and no new safety signals were observed. CONCLUSIONS: Mature PFS data from ALEX confirmed significant improvement in PFS for alectinib over crizotinib in ALK-positive NSCLC. OS data remain immature, with a higher 5-year OS rate with alectinib versus crizotinib. This is the first global randomized study to show clinically meaningful improvement in OS for a next-generation tyrosine kinase inhibitor versus crizotinib in treatment-naive ALK-positive NSCLC. CLINICAL TRIALS NUMBER: NCT02075840.

Interaction of the Gulf Stream with small scale topography: a focus on lee waves.

The generation of lee waves in the Gulf Stream along the U.S. seaboard is investigated using high resolution realistic simulations. The model reproduces the surface signature of the waves, which compares favourably with observations from satellite sun glitter images in the region. In particular, a large number of internal waves are observed above the Charleston Bump. These waves match well with the linear theory describing topographically-generated internal waves, which can be used to estimate the associated vertical transport of momentum and energy extracted from the mean flow. Finally, small scale topographic features are shown to have a significant impact on the mean flow in this region of the Gulf Stream, and the specific role of lee waves in this context is outlined.

Oral squamous carcinoma cells promote macrophage polarization in an MIF-dependent manner.

BACKGROUND: Tumor-associated macrophages (TAMs) are important determinants of intratumoral immune evasion, neoangiogenesis, extracellular matrix remodeling and dysregulated tumor cell proliferation. Our prior studies revealed that macrophage-derived, but not tumor cell-derived, macrophage migration inhibitory factor (MIF), is an important determinant of TAM alternative activation and M2 polarization. AIM: Because MIF is historically thought to initiate signaling via a receptor-dependent, outside-in mode of action, we wished to investigate the specific contributions of tumor-derived vs. macrophage-derived MIF to M2 marker expression during macrophage polarization. DESIGN: Murine oral squamous cell-carcinoma cells (SCCVII) were co-cultured with either the RAW 264.7 mouse macrophage cell line or mouse primary bone marrow-derived macrophages in the context of MIF genetic loss/inhibition individually or in combination each cell type. METHODS: Twelve well Transwell plates were used to co-culture SCCVII cells and RAW 264.7, MIF+/+ or MIF-/- macrophages treated with/without the small molecule MIF inhibitor, 4-iodo-6-phenylpyrimidine and incubated in the presence or absence of interleukin (IL-4) for 48 h. Macrophages were analyzed by quantitative real-time polymerase chain reaction and/or immunoblotting for relative macrophage polarization marker expression. RESULTS: IL-4 treatment synergizes with SCCVII co-culture in inducing the expression of macrophage M2 markers and loss or inhibition of macrophage-derived MIF significantly reduces both IL-4 alone and IL-4/SCCVII co-culture-induced macrophage M2 marker expression. CONCLUSION: These studies identify an important and dominant requirement for macrophage MIF in maximal Th2-cytokine and oral squamous carcinoma cell-induced macrophage polarization and M2 marker expression.

Improving the physical properties of starch using a new kind of water dispersible nano-hybrid reinforcement.

Plasticized cassava starch matrix composites reinforced by a multi-wall carbon nanotube (MWCNT)-hercynite (FeAl2O4) nanomaterial were developed. The hybrid nanomaterial consists of FeAl2O4 nanoparticles anchored strongly to the surface of the MWCNT. This nano-hybrid filler shows an irregular geometry, which provides a strong mechanical interlocking with the matrix, and excellent stability in water, ensuring a good dispersion in the starch matrix. The composite containing 0.04wt.% of the nano-hybrid filler displays increments of 370% in the Young's modulus, 138% in tensile strength and 350% in tensile toughness and a 70% decrease in water vapor permeability relative to the matrix material. All of these significant improvements are explained in terms of the nano-hybrid filler homogenous dispersion and its high affinity with both plasticizers, glycerol and water, which induces crystallization without deterioration of the tensile toughness.

[Nursing homes versus assisted living facilities: Outcome quality regarding pressure ulcers, falls and malnutrition]. / Vollstationäre Pflegeeinrichtungen vs. "betreutes Wohnen mit ambulanter Versorgung": Ergebnisqualität bezogen auf Dekubitus, Sturz und Mangelernährung.

BACKGROUND: Assisted living has become more popular among individuals who are care-dependent or are threatened with care dependency as an attractive alternative to a standard nursing home. As differences regarding nursing care quality in both settings are so far unknown, the aim of the study was to compare the quality indicators occurrence of pressure ulcers, falls and malnutrition in both settings. MATERIAL AND METHODS: In spring 2010 a multicentre cross-sectional study on the prevalence of care problems was conducted including 3610 individuals in 31 nursing homes (NH) and 21 assisted living facilities (ALF) following a standardized study protocol. Comparative analyses of raw and adjusted prevalence of the care problems occurrence of pressure ulcers, falls and malnutrition were conducted. RESULTS: Individuals in the ALFs were on average older than those in the NHs (89.5 years vs. 83.2 years) but were more mobile and less care-dependent. The prevalence of care problems in the NHs compared to those in the ALFs showed the following results: pressure ulcers (NH = 3.9%, ALF = 2.3%), nosocomial pressure ulcer prevalence (NH = 2.3%, ALF = 0.2%), incidence of falls within 3 months (NH = 12.5%, ALF = 20.4%) and malnutrition according to a body mass index (BMI) ≤ 18.5 kg/m(2) (NH = 5.6%, ALF = 11.4%). CONCLUSION: Due to the population differences, no final conclusions about the quality of care according to the analysed indicators can be made. While the prevalence of pressure ulcers was remarkably lower, falls and malnutrition had a higher prevalence in ALFs. Ongoing and systematic outcome quality surveys in different long-term care facilities are recommended.

Cardiovascular results of an individually controlled complex prevention.

UNLABELLED: Lifestyle modifications (increased level of physical activity, favourable nutrition, and stress management) are important factors in the prevention of and the therapy for cardiovascular (CV) diseases. OBJECTIVES: The effects of an individualized, half-year long exercise program on CV risk factors were investigated in 50 patients with moderately high CV risk factors. PATIENTS AND METHODS: 75 subjects participated in the study. After the eleventh week of regular training, members of Group A performed 55-65 minutes of exercise at 4-5 times a week, while patients in Group B took part in 45-55 minute training sessions at 2-3 times a week. Activities were monitored using POLAR devices and controlled by a cardiologist and an exercise training expert. Members of the control group (C)were also affected by risk factors, they, however, were not involved in any physical activity. RESULTS: A marked improvement was seen in performance level (62% in Group A, 38% in Group B). There was a decrease in the LDL cholesterol level (30% and 21%), total cholesterol (16% and 14%), triglyceride (23% in both groups), and an increase in the HDL-cholesterol level (53% and 26%). Body mass (BM) decreased in both groups (8.7% and 5%).In addition, a decrease was also seen in the resting heart rate (HR) (9.6% and 4.5%) and blood pressure (BP: systolic 8.5% and 5.5 %, diastolic 7% and 4.7%). CONCLUSIONS: In persons affected by CV risk factors, lifestyle modification with personal, HR controlled complex (cardio and resistance) aerobic training effectively decreased CV risk factors and strongly improved state of health and quality of life.

Incidence and Pathogenicity of Plant-Parasitic Nematodes Associated with Blueberry (Vaccinium spp.) Replant Disease in Georgia and North Carolina.

Blueberry replant disease (BRD) is an emerging threat to continued blueberry (Vaccinium spp.) production in Georgia and North Carolina. Since high populations of ring nematode Mesocriconema ornatum were found to be associated with commercially grown blueberries in Georgia, we hypothesized that M. ornatum may be responsible for predisposing blueberry to BRD. We therefore tested the pathogenicity of M. ornatum on 10-wk-old Rabbiteye blueberries (Vaccinium virgatum) by inoculating with initial populations (Pi) of 0 (water control), 10, 100, 1,000. and 10,000 mixed stages of M. ornatum/pot under both greenhouse (25 ± 2°C) and field microplot conditions. Nematode soil population densities and reproduction rates were assessed 75, 150, 225, and 255, and 75, 150, 225, and 375 d after inoculation (DAI) in both the greenhouse and field experiments, respectively. Plant growth parameters were recorded in the greenhouse and field microplot experiments at 255 and 375 DAI, respectively. The highest M. ornatum population density occurred with the highest Pi level, at 75 and 150 DAI under both greenhouse (P < 0.01) and field (P < 0.01) conditions. However, M. ornatum rate of reproduction increased significantly in pots receiving the lowest Pi level of 10 nematodes/plant compared with the pots receiving Pi levels of 100, 1,000, and 10,000 nematodes 75 DAI. Plant-parasitic nematode populations were determined in commercial blueberry replant sites in Georgia and North Carolina during the 2010 growing season. Mesocriconema ornatum and Dolichodorus spp. were the predominant plant-parasitic nematodes in Georgia and North Carolina, respectively, with M. ornatum occurring in nearly half the blueberry fields sampled in Georgia. Other nematode genera detected in both states included Tylenchorhynchus spp., Hoplolaimus spp., Hemicycliophora spp., and Xiphinema spp. Paratrichodorus spp. was also found only in Georgia. In Georgia, our results indicate that blueberry is a host for M. ornatum and its relationship to BRD warrants further investigation.

Development and characterization of starch nanoparticles by gamma radiation: potential application as starch matrix filler.

Gamma radiation arises as an advantageous alternative to obtain starch nanoparticles given its low cost, simple methodology and scalability. Starch nanoparticles (SNP) with sizes around 20 and 30 nm were obtained applying a dose of 20 kGy from cassava (CNP-γ) and waxy maize (WNP-γ) starch, respectively. They showed the same thermal degradation behavior and their maximum mass loss zone was similar to those nanoparticles obtained from acid hydrolysis (WNP-h). Additionally, CNP-γ and WNP-γ were used as nanofillers in a cassava matrix. Increments of 102% in storage modulus were obtained with the addition of only 2.5 wt.% of WNP-γ, showing that gamma radiation is a successful methodology to obtain SNP able to be used as starch reinforcement.

Toxicity of endophyte-infected tall fescue alkaloids and grass metabolites on Pratylenchus scribneri.

ABSTRACT Neotyphodium coenophialum, an endophytic fungus associated with tall fescue grass, enhances host fitness and imparts pest resistance. This symbiotum is implicated in the reduction of stresses, including plant-parasitic nematodes. To substantiate this implication, toxicological effects of root extracts, polyphenolic fraction, ergot, and loline alkaloids from endophyte-infected tall fescue were investigated using Pratylenchus scribneri, a nematode pest of tall fescue. In vitro bioassays and greenhouse studies were used as tests for effects of root fractions and compounds on motility and mortality of this lesion nematode. Greenhouse studies revealed that endophyte-infected tall fescue grasses are essentially nonhosts to P. scribneri, with root populations averaging 3 to 17 nematodes/pot, compared with 4,866 and 8,450 nematodes/pot for noninfected grasses. The in vitro assay indicated that root extracts from infected tall fescues were nematistatic. Polyphenols identified in extracts included chlorogenic acid, 3,5-dicaffeoylquinic acids, caffeic acid, and two unidentified compounds, but these were not correlated with endophyte status, qualitatively or quantitatively. Tests of several ergot alkaloids revealed that ergovaline and alpha-ergocryptine were nematicidal at 5 and 50 microg/ml, respectively, while ergocornine and ergonovine were nematistatic at most concentrations. Loline (N-formylloline), the pyrrolizidine alkaloid tested, was nematicidal (50 to 200 microg/ml). The ecological benefits of the metabolites tested here should assist in defining their role in deterring this nematode species while offering some probable mechanisms of action against plant-parasitic nematodes in general.

Endosymbiotic bacteria in the parasitic ciliate Ichthyophthirius multifiliis.

Endosymbiotic bacteria were identified in the parasitic ciliate Ichthyophthirius multifiliis, a common pathogen of freshwater fish. PCR amplification of DNA prepared from two isolates of I. multifiliis, using primers that bind conserved sequences in bacterial 16S rRNA genes, generated an approximately 1,460-bp DNA product, which was cloned and sequenced. Sequence analysis demonstrated that 16S rRNA gene sequences from three classes of bacteria were present in the PCR product. These included Alphaproteobacteria (Rickettsiales), Sphingobacteria, and Flavobacterium columnare. DAPI (4',6-diamidino-2-phenylindole) staining showed endosymbionts dispersed throughout the cytoplasm of trophonts and, in most, but not all theronts. Endosymbionts were observed by transmission electron microscopy in the cytoplasm, surrounded by a prominent, electron-translucent halo characteristic of Rickettsia. Fluorescence in situ hybridization demonstrated that bacteria from the Rickettsiales and Sphingobacteriales classes are endosymbionts of I. multifiliis, found in the cytoplasm, but not in the macronucleus or micronucleus. In contrast, F. columnare was not detected by fluorescence in situ hybridization. It likely adheres to I. multifiliis through association with cilia. The role that endosymbiotic bacteria play in the life history of I. multifiliis is not known.

Characterization of the mouse bile salt export pump overexpressed in the baculovirus system.

The bile salt export pump (Bsep), a member of the ATP-binding cassette superfamily of transporters, mediates the ATP-dependent canalicular secretion of bile salts. We have cloned and expressed the mouse Bsep (mBsep) protein in Sf9 insect cells, and characterized its transport and ATPase properties. Because its deduced amino acid sequence predicts multiple phosphorylation sites for protein kinase A, protein kinase C (PKC) and Ca(2+)-calmodulin dependent kinase II, we have also tested whether mBsep undergoes phosphorylation. MBsep transports both glycine and taurine conjugated bile salts. Sf9 cell membranes that express mBsep exhibit higher basal ATPase activity than control membranes, and this is further stimulated by bile salts and inhibited by vanadate. Taurochenodeoxycholate is transported with the highest affinity and is the most potent inducer of ATPase activity. Cyclosporin A, glibenclamide and rifamycin SV, all competitive inhibitors of Bsep transport, also reduced the bile salt-stimulated ATPase activity. MBsep exists as a phospho-protein when expressed in Sf9 cells and the immunoprecipitated mBsep complex is a substrate for the catalytic subunit of PKC. When mBsep and the alpha-isoform of mouse PKC are co-expressed in Sf9 cells, a ninefold stimulation of phosphorylation occurs. This is further increased to 18-fold after activation by phorbol ester. Given that bile salts activate selected PKC isoforms in hepatocytes, including the alpha isoform, the phosphorylation of mBsep by PKCalpha may represent a point of regulation for this transporter that is mediated by its own substrate.

Cross-talk between olfactory second messenger pathways.

The second messengers 3'-5'-cyclic-monophosphate (cAMP) and inositol 1,4,5-trisphosphate (InsP3) have been implicated in olfactory signal transduction in various species. The results of the present study provide evidence that the two olfactory second messenger pathways in rat olfactory neurons do not work independently but rather show a functional antagonism: whereas inhibition of phospholipase C (PLC) in isolated olfactory cilia by U-73122 led to an augmentation of odor-induced cAMP signaling, activation of the phosphoinositol pathway resulted in attenuation of odor-induced cAMP formation. Furthermore, this study indicates that elevated cAMP levels cause suppression of odor-induced InsP3 signaling, whereas inhibition of adenylate cyclase (AC) by cisN-(2-phenylcyclopentyl)azacylotridec-1-en-2-amine (MDL-12,330 A) results in potentiation of odor-induced InsP3 formation. Concerning the molecular mechanism involved in cross-interaction, the experimental data indicate that the observed antagonism of elevated cAMP is based on inhibition of PLC activation rather than on stimulation of InsP3 degradation. As blockage of the endogenous protein kinase A (PKA) prevented the inhibitory effect of cAMP, the suppression of odor-induced InsP3 signaling by cAMP may be mediated by a PKA-controlled reaction.

Chicken Litter Soil Amendment Effects on Soilborne Microbes and Meloidogyne incognita on Cotton.

The effects of chicken litter incorporated 28, 14, and 0 days before planting on Meloidogyne incognita in cotton and soil organisms were determined in the greenhouse. Treatments consisted of field soil amended with litter at rates of 0, 0.125, 0.25, 0.5, and 1% by weight. At 45 and 90 days after planting, numbers of M. incognita decreased as rates of litter increased. Microbivorous nematode densities increased as litter rates increased only in the first experiment. Plant growth increased as litter rates increased, regardless of when the litter was incorporated, or the presence or absence of M. incognita. Bacterial and fungal CFU fluctuated during both experiments, but generally had positive linear relationships with litter rate. Population densities of M. incognita decreased with increasing bacterial and fungal counts in amended soil. Bacterial genera identified from the litter-amended soil included Arthrobacter, Bacillus, Cellulomonas, Mi-crococcus, Pseudomonas, and Rhodococcus.

Functional and molecular characterization of individual olfactory neurons.

To gain an understanding of the olfactory signal transduction process, individual chemosensory neurons have been assessed for odor-induced Ca2+ responses and the molecular elements of transduction cascades using Ca2+ imaging technique in combination with single-cell RT-PCR approaches. It has been demonstrated that responsiveness of cells to cyclic AMP or inositol trisphosphate odorants was blocked by specific adenylyl cyclase inhibitors or phospholipase C inhibitors, respectively. Using specific primers in single-cell RT-PCR analysis, olfactory marker protein, two G protein subtypes (G(olf) and G(o)), and adenylyl cyclase (subtype III) and a phospholipase C (phospholipase Cbeta2-related subtype) were identified. For a subpopulation of sensory neurons it was demonstrated that both transduction cascades coexist and are active in the same cell. These data support the notion that two second messenger pathways are active in olfactory sensory neurons and emphasize the concept of dual transduction cascades in olfaction.

Identification of a phospholipase C beta subtype in rat taste cells.

From rat circumvallate papillae a novel phospholipase C (PLC) subtype has been cloned and identified as most closely related to human PLC beta2. The corresponding mRNA was only detected in sensory lingual tissue but not in non-taste lingual tissue or any other tissues examined by Northern blot analysis. In situ hybridization revealed that a subset of taste receptor cells of circumvallate papillae was specifically labeled. A functional involvement of this PLC beta subtype in taste signal transduction emerged from biochemical analysis monitoring the second messenger response in circumvallate preparations induced by denatonium benzoate. This bitter agent elicited a rapid and transient increase of the inositol 1,4,5-trisphosphate level; this response was blocked by U73122, a potent inhibitor of PLC, and by PLC beta2-specific antibodies. These data indicate that a phospholipase C beta2 isoform mediates a denatonium benzoate-induced second messenger response of taste sensory cells in the circumvallate papillae.

Sodium/calcium exchanger in rat olfactory neurons.

The chemo-electrical transduction process in olfactory neurons is accompanied by a rapid and transient increase in intracellular calcium concentrations. The notion that Na+/Ca2+ exchanger activities may play a major role in extruding calcium ions out of the cell and maintaining Ca2+ homeostasis in olfactory receptor cells was assessed by means of laser scanning confocal microscopy in combination with the fluorescent indicators Fluo-3 and Fura-Red. The data indicate that high exchanger activity, which was inhibited by amiloride derivatives, is located in the dendritic knob and probably in the olfactory cilia. This result was supported by experiments using specific antiserum raised against retinal Na+/Ca2+ exchanger protein which labelled an immunoreactive protein of 230 kDa in Western blots from olfactory tissue and strongly stained the ciliary layer of the olfactory epithelium.

Meloidogyne incognita Infested Soil Amended With Chicken Litter.

The effects of chicken litter on Meloidogyne incognita in cotton, Gossypium hirsutum cv. DPL50 were determined in field microplots. Litters (manure and pine-shaving bedding) from a research facility and a commercial broiler house were used. Treatments consisted of 0.25%, 0.5%, and 1% litter by dry weight of soil for each kind of litter. Three control treatments consisted of soil not amended with litter, with and without nematodes, and one treatment to which mineral fertilizer was added at a nitrogen rate equivalent to that of the 0.5% litter rate, with nematodes. Microplots were inoculated at planting with 900 eggs/100 cm(3) soil in 1993 and 1,000 eggs/100 cm(3) soil in 1994. At 92 and 184 days after planting, nematode population densities decreased linearly with increasing rates of litter. Nematode numbers at midseason were larger in plots treated with mineral fertilizer than in plots treated with a rate of litter equivalent to the 0.5% rate. Fungal and bacterial population densities fluctuated throughout the growing season. Bacterial numbers had a positive linear relationship, with increasing rates of litter only in October 1993; however, significant positive relationships were observed throughout the 1994 growing season. In 1994, nematode population density at 92 days after planting decreased linearly with increasing bacterial numbers 30 days after planting. No other significant relationships between nematode densities and microbial densities were observed. Fungi and bacteria isolated from the litter and litter-amended soil were identified. Fungal genera isolated included Acremonium, Aspergillus, Eurotium, Paecilomyces, Petriella, and Scopulariopsis, whereas bacteria genera included Arthrobacter, Bacillus, and Pseudomonus.

Damage Potential and Reproduction of Meloidogyne incognita Race 3 and M. arenaria Race 1 on Kenaf.

The effects of Meloidogyne incognita race 3 and M. arenaria race 1 on growth of kenaf cv. Everglades 41 was determined under greenhouse conditions. Seedlings of kenaf were inoculated with initial population densities (Pi) of 0, 625, 1,250, 2,500, 5,000, and 10,000 eggs/plant and placed on greenhouse benches in a randomized complete block design. Plant growth and nematode reproduction were assessed 6 and 12 weeks after inoculation. Growth suppression of kenaf in response to increasing Pi was observed 6 weeks after inoculation. Severe damage was observed by all Pi levels of M. incognita and M. arenaria at 12 weeks after inoculation. Plant height, basal stem diameter, and fresh and dry shoot weights had a negative linear relationship to log(Pi + 1) for both M. incognita and M. arenaria. Plant height was reduced 25%, basal stem diameter was reduced 19%, and dry shoot weights were reduced 64% at the highest inoculum rates of M. incognita. Similar reductions were observed for M. arenaria. Greater levels of root necrosis were observed with M. incognita than M. arenaria at similar inoculum levels. High reproductive factors (Rf) were observed on kenaf for both M. incognita (48 - 1,804) and M. arenaria (257 - 4,240), with the highest Rf values occurring at the lowest Pi. The host status of kenaf renders it unsuitable for use in rotation systems with other susceptible crops.