Cutting edge: Slamf8 is a negative regulator of Nox2 activity in macrophages

Slamf8 (CD353) is a cell surface receptor that is expressed upon activation of macrophages (MΦs) by IFN-γ or bacteria. In this article, we report that a very high NADPH oxidase (Nox2) enzyme activity was found in Slamf8(-/-) MΦs in response to Escherichia coli or Staphylococcus aureus, as well as to PMA. The elevated Nox2 activity in Slamf8(-/-) MΦs was also demonstrated in E. coli or S. aureus phagosomes by using a pH indicator system and was further confirmed by a reduction in the enzyme activity after transfection of the receptor into Slamf8-deficient primary MΦs or RAW 264.7 cells. Upon exposure to bacteria or PMA, protein kinase C activity in Slamf8(-/-) MΦs is increased. This results in an enhanced phosphorylation of p40phox, one key component of the Nox2 enzyme complex, which, in turn, leads to greater Nox2 activity. Taken together, the data show that, in response to inflammation-associated stimuli, the inducible receptor Slamf8 negatively regulates inflammatory responses.     

New sources of resistance to lettuce aphids in Lactuca spp.

The use of resistant cultivars is one of the best ways to protect lettuce from aphid pests. At present, there are cultivars available with nearly complete resistance to Nasonovia ribisnigri biotype Nr:0 (based on the Nr gene) and partial resistance to Macrosiphum euphorbiae. Nevertheless, a new biotype of N. ribisnigri (Nr:1) able to overcome the resistance based on the Nr gene is expanding around Europe and has become a major threat of lettuce. In the present work, we report the presence of this new biotype in southeastern Spain, a major lettuce-producing region. Furthermore, a pool of 264 germplasm accessions belonging to Lactuca genus was tested in a greenhouse assay to search for new resistance sources to N. ribisnigri. The most promising accessions were retested in the laboratory to characterize the resistance by means of free-choice and antibiosis assays against biotypes Nr:0 and Nr:1 of N. ribisnigri and against a clone of M. euphorbiae. Three accessions of L. virosa showed resistance against the target aphid species and could be of interest for ongoing breeding programs. The accessions CGN16272 and CGN13361 were both partially resistant to the Nr:1 biotype of N. ribisnigri and to M. euphorbiae, and CGN13355, in spite of not being resistant to N. ribisnigri, showed a near complete resistance to M. euphorbiae. The study of the feeding behavior of N. ribisnigri biotypes showed that the Nr:1 biotype is able to maintain a similar phloem feeding ingestion pattern on genotypes bearing the Nr gene and on N. ribisnigri-susceptible lettuce genotypes. Moreover, as aphids rejected L. virosa as a feeding source due to superficial factors (high level of antixenosis), no differences in the level of antibiosis between such genotypes were detected. A second set of screening assays were conducted on 40 accessions of L. virosa in order to select for resistance against the Nr:1 biotype. The results showed three accessions with high levels of resistance (CGN05148, CGN21399 and CGN16274) against Nr:1 that could be of interest in lettuce breeding programs.     

Prognostic Impact of del(17p) and del(22q) as Assessed by Interphase FISH in Sporadic Colorectal Carcinomas

Background

Most sporadic colorectal cancer (sCRC) deaths are caused by metastatic dissemination of the primary tumor. New advances in genetic profiling of sCRC suggest that the primary tumor may contain a cell population with metastatic potential. Here we compare the cytogenetic profile of primary tumors from liver metastatic versus non-metastatic sCRC.

Methodology/Principal Findings

We prospectively analyzed the frequency of numerical/structural abnormalities of chromosomes 1, 7, 8, 13, 14, 17, 18, 20, and 22 by iFISH in 58 sCRC patients: thirty-one non-metastatic (54%) vs. 27 metastatic (46%) disease. From a total of 18 probes, significant differences emerged only for the 17p11.2 and 22q11.2 chromosomal regions. Patients with liver metastatic sCRC showed an increased frequency of del(17p11.2) (10% vs. 67%;p<.001) and del(22q11.2) (0% vs. 22%;p = .02) versusnon-metastatic cases. Multivariate analysis of prognostic factors for overall survival (OS) showed that the only clinical and cytogenetic parameters that had an independent adverse impact on patient outcome were the presence of del(17p) with a 17p11.2 breakpoint and del(22q11.2). Based on these two cytogenetic variables, patients were classified into three groups: low- (no adverse features), intermediate- (one adverse feature) and high-risk (two adverse features)- with significantly different OS rates at 5-years (p<.001): 92%, 53% and 0%, respectively.

Conclusions/Significance

Our results unravel the potential implication of del(17p11.2) in sCRC patients with liver metastasis as this cytogenetic alteration appears to be intrinsically related to an increased metastatic potential and a poor outcome, providing additional prognostic information to that associated with other cytogenetic alterations such as del(22q11.2). Additional prospective studies in larger series of patients would be required to confirm the clinical utility of the new prognostic markers identified.     

Usefulness of truncal obesity indices as predictive factors for obstructive sleep apnea syndrome

Background: Optimization of the indications for nocturnal polysomnography in the diagnosis of obstructive sleep apnea syndrome (OSAS) could lead to significant reductions in health expenditure. To this end, we assessed the usefulness of truncal obesity measurements in the diagnosis of OSAS. Methods and Procedures: One hundred ninety‐two patients (152 men and 40 women) referred to our clinic with suspected OSAS underwent a complete polysomnography study and their BMI and truncal obesity measurements were obtained. Results: Comparison of patients defined as snorers with OSAS patients, by means of the Student's t ‐test revealed significant differences in the truncal obesity parameters such as waist‐to‐hip ratio (0.94 vs. 0.98) and waist circumference (100.7 cm vs. 106.3 cm). We found no significant differences between BMI values in the two groups. Indices of truncal obesity were analyzed as predictive factors for OSAS using a multivariate logistic regression model that included variable sex, and in which waist‐to‐hip ratio, BMI, neck circumference, and age were analyzed as binary variables. According to this model, our patients' risk of suffering from OSAS was 2.6 times greater if the waist‐to‐hip ratio was >1 in men and >0.85 in women. The risk of OSAS was also increased if the patients were men or over 52 years of age. Discussion: Obtaining simple measurements such as those for truncal obesity may help prioritize the use of polysomnography in patients with a greater risk of OSAS. Our results suggest that BMI is not a good predictor of OSAS in a group of patients with a high BMI.     

Comparative studies on the conformational change and aggregation behavior of irradiated carrageenans and agar by dynamic light scattering

The conformational associative properties of kappa-, iota-, and lambda-carrageenan and agar with irradiation dose were studied by dynamic light scattering. The random scission of the carrageenans and agar by gamma irradiation resulted in the formation of polydispersed lower molecular weight fragments. At high doses, the system moves towards uniformity. Conformational change from coil to helix was observed in all carrageenans and agar at doses up to 100 kGy. The conformational change in lambda-carrageenan may be due to the irregular and hybrid structure of this polysaccharide. Only agar and lambda-carrageenan still undergo conformational transition at a high dose of 200 kGy. Gelation is observed for kappa-, iota-carrageenan up to a dose of 50 kGy while gelation is still observed at 100 kGy for agar. Increase in the hydrodynamic radius with decreasing temperatures for the non-irradiated carrageenans follows this order: lambda-carrageenan>kappa-carrageenan>iota-carrageenan. Slight increases in hydrodynamic radius were observed with irradiation.     

Discovery of novel 4-amino-6-arylaminopyrimidine-5-carbaldehyde oximes as dual inhibitors of EGFR and ErbB-2 protein tyrosine kinases

We herein disclose a novel series of 4-aminopyrimidine-5-carbaldehyde oximes that are potent and selective inhibitors of both EGFR and ErbB-2 tyrosine kinases, with IC(50) values in the nanomolar range. Structure-activity relationship (SAR) studies elucidated a critical role for the 4-amino and C-6 arylamino moieties. The X-ray co-crystal structure of EGFR with 37 was determined and validated our design rationale.     

Trichomes and photosynthetic pigment composition changes: responses of <Emphasis Type="Italic">Quercus ilex </Emphasis>subsp. <Emphasis Type="Italic">ballota</Emphasis> (Desf.) Samp. and<Emphasis Type="Italic"> Quercus coccifera</Emphasis> L. to Mediterranean stress conditions

Sun and shade leaves of two Mediterranean Quercus species, Quercus ilex subsp. ballota (Desf.) Samp. and Quercus coccifera L., were compared by measuring leaf optical properties, photosynthetic pigment composition and photosystem II efficiency. The presence of trichomes in the adaxial (upper) leaf surface of Q. ilex subsp. ballota seems to constitute an important morphological mechanism that allows this species to maintain a good photosystem II efficiency during the summer. Q. coccifera has almost no trichomes and seems instead to develop other physiological responses, including a smaller light-harvesting antenna size, higher concentrations of violaxanthin cycle pigments and a higher (zeaxanthin + antheraxanthin)/(violaxanthin + antheraxanthin + zeaxanthin) ratio. Q. coccifera was not able to maintain a good photosystem II efficiency up to the end of the summer. In Q. ilex subsp. ballota leaves, natural loss or mechanical removal of adaxial-face leaf trichomes induced short-term decreases in photosystem II efficiency. These changes were accompanied by de-epoxidation of violaxanthin cycle pigments, suggesting that the absence of trichomes would trigger physiological responses in this species. Our data have revealed different patterns of response of Q. ilex subsp. ballota and Q. coccifera facing the stress conditions prevailing in the Mediterranean area.     

The amphipathic peptide mellitin as a tool to study the membrane-dependent activation of tissue transglutaminase

Summary The role of membrane phospholipids on the cross-linking activity of guinea pig liver (tissue) transglutaminase has been investigated using the amphipathic model peptide melittin as glutaminyl substrate and the primary amine monodansylcadaverine as extrinsic amine donor. A marked increase of transglutaminase catalytic activity was observedin vitro assays in the presence of neutral membrane phospholipids. In contrast, activation was abolished in the presence of membranes containing pure anionic lipids. Enzyme activation could be ascribed to a direct binding of the lipid to the protein as demonstrated in enzymatic assays using a non membrane-interacting peptide (Cbz-Gln-Gly). The data obtained with model peptides suggest that the cross-linking activity of tissue transglutaminase could be modulated by the local microenvironment composition of the lipid bilayer and indicate that membrane phospholipids should be taken into account for further experiments directed to asess, the still poor understood, physiological role of tissue transglutaminase.