Multiploidy occurrence in tomato calli from anther culture

Anther culture has long been used for the production of fully homozygous lines in order to produce, mainly, doubled haploid plants, which are of great interest in plant breeding. For tomato, a recalcitrant species for androgenesis production protocols have not been standardized. It is known that the genotype, anther size, the developmental stage of the microspore, and the medium composition are some factors that can influence the calli production. The present study aimed to adapt flow cytometry methodology to verify the microsporogenesis phases of anthers in order to assess the anther responsiveness of different tomato genotypes in an androgenesis-induction culture medium and to analyze the DNA ploidy level of calli produced by flow cytometry. Anthers from flower buds of length 1.0 to 5.9 mm, corresponding to the size range as analyzed by flow cytometry and cytogenetic methods, were inoculated into Murashige and Skoog (MS) basal medium containing the growth regulators 6-(y,ydimethylallylamino) purine and indole-3-acetic acid. The obtained calli were subsequently analyzed by flow cytometry to determine the DNA ploidy level. Surprisingly, despite no pretreatment with microtubule-depolymerizing agents, five classes of multiploid calli were observed, as follows: class I (2C-4C-8C-16C), class II (2C-4C-8C-16C-32C), class III (4C-8C), class IV (4C-8C-16C) and class V (8C-16C- 32C). Multiploid calli were identified in short-term (two month) culturing, suggesting that the variable culture duration did not directly influence the occurrence of endoreduplication. In this work, this type of somaclonal variation has been reported for the first time in tomato anther culture, and their possible origin has been discussed.Key words: Callogenesis, flow cytometry, polyploidy, Solanum lycopersicum, somaclonal variation

Antiinflammatory Therapy with Canakinumab for Atherosclerotic Disease

Background: Experimental and clinical data suggest that reducing inflammation without affecting lipid levels may reduce the risk of cardiovascular disease. Yet, the inflammatory hypothesis of atherothrombosis has remained unproved. Methods: We conducted a randomized, double-blind trial of canakinumab, a therapeutic monoclonal antibody targeting interleukin-1β, involving 10,061 patients with previous myocardial infarction and a high-sensitivity C-reactive protein level of 2 mg or more per liter. The trial compared three doses of canakinumab (50 mg, 150 mg, and 300 mg, administered subcutaneously every 3 months) with placebo. The primary efficacy end point was nonfatal myocardial infarction, nonfatal stroke, or cardiovascular death. RESULTS: At 48 months, the median reduction from baseline in the high-sensitivity C-reactive protein level was 26 percentage points greater in the group that received the 50-mg dose of canakinumab, 37 percentage points greater in the 150-mg group, and 41 percentage points greater in the 300-mg group than in the placebo group. Canakinumab did not reduce lipid levels from baseline. At a median follow-up of 3.7 years, the incidence rate for the primary end point was 4.50 events per 100 person-years in the placebo group, 4.11 events per 100 person-years in the 50-mg group, 3.86 events per 100 person-years in the 150-mg group, and 3.90 events per 100 person-years in the 300-mg group. The hazard ratios as compared with placebo were as follows: in the 50-mg group, 0.93 (95% confidence interval [CI], 0.80 to 1.07; P = 0.30); in the 150-mg group, 0.85 (95% CI, 0.74 to 0.98; P = 0.021); and in the 300-mg group, 0.86 (95% CI, 0.75 to 0.99; P = 0.031). The 150-mg dose, but not the other doses, met the prespecified multiplicity-adjusted threshold for statistical significance for the primary end point and the secondary end point that additionally included hospitalization for unstable angina that led to urgent revascularization (hazard ratio vs. placebo, 0.83; 95% CI, 0.73 to 0.95; P = 0.005). Canakinumab was associated with a higher incidence of fatal infection than was placebo. There was no significant difference in all-cause mortality (hazard ratio for all canakinumab doses vs. placebo, 0.94; 95% CI, 0.83 to 1.06; P = 0.31). Conclusions: Antiinflammatory therapy targeting the interleukin-1β innate immunity pathway with canakinumab at a dose of 150 mg every 3 months led to a significantly lower rate of recurrent cardiovascular events than placebo, independent of lipid-level lowering. (Funded by Novartis; CANTOS ClinicalTrials.gov number, NCT01327846.

Long-term outcomes of clinical complete responders after neoadjuvant treatment for rectal cancer in the International Watch & Wait Database (IWWD): an international multicentre registry study

Background: The strategy of watch and wait (W&W) in patients with rectal cancer who achieve a complete clinical response (cCR) after neoadjuvant therapy is new and offers an opportunity for patients to avoid major resection surgery. However, evidence is based on small-to-moderate sized series from specialist centres. The International Watch & Wait Database (IWWD) aims to describe the outcome of the W&W strategy in a large-scale registry of pooled individual patient data. We report the results of a descriptive analysis after inclusion of more than 1000 patients in the registry. Methods: Participating centres entered data in the registry through an online, highly secured, and encrypted research data server. Data included baseline characteristics, neoadjuvant therapy, imaging protocols, incidence of local regrowth and distant metastasis, and survival status. All patients with rectal cancer in whom the standard of care (total mesorectal excision surgery) was omitted after neoadjuvant therapy were eligible to be included in the IWWD. For the present analysis, we only selected patients with no signs of residual tumour at reassessment (a cCR). We analysed the proportion of patients with local regrowth, proportion of patients with distant metastases, 5-year overall survival, and 5-year disease-specific survival. Findings: Between April 14, 2015, and June 30, 2017, we identified 1009 patients who received neoadjuvant treatment and were managed by W&W in the database from 47 participating institutes (15 countries). We included 880 (87%) patients with a cCR. Median follow-up time was 3·3 years (95% CI 3·1–3·6). The 2-year cumulative incidence of local regrowth was 25·2% (95% CI 22·2–28·5%), 88% of all local regrowth was diagnosed in the first 2 years, and 97% of local regrowth was located in the bowel wall. Distant metastasis were diagnosed in 71 (8%) of 880 patients. 5-year overall survival was 85% (95% CI 80·9–87·7%), and 5-year disease-specific survival was 94% (91–96%). Interpretation: This dataset has the largest series of patients with rectal cancer treated with a W&W approach, consisting of approximately 50% data from previous cohort series and 50% unpublished data. Local regrowth occurs mostly in the first 2 years and in the bowel wall, emphasising the importance of endoscopic surveillance to ensure the option of deferred curative surgery. Local unsalvageable disease after W&W was rare. Funding: European Registration of Cancer Care financed by European Society of Surgical Oncology, Champalimaud Foundation Lisbon, Bas Mulder Award granted by the Alpe d'Huzes Foundation and Dutch Cancer Society, and European Research Council Advanced Grant

Diversity of gall-inducing insects in the high altitude wetland forests in Pernambuco, Northeastern Brazil

We report on the richness of galling insects in the altitudinal wetland forests of Pernambuco State, Northeastern Brazil. We found 80 distinct types of insect galls on 49 species of host plants belonging to 28 families and 35 genera. Most of the galled plant species belong to Nyctaginaceae, Fabaceae, Meliaceae, Sapindaceae and Myrtaceae. The most common gall were spheroid and globoid; most galls were glabrous, predominantly green and with one chamber, and on the leaves. Most galls were induced by Cecidomyiidae (Diptera). The results of this study contribute to existing knowledge richness of galling insects and host-plant diversity in the altitudinal wetland forests of Northeastern Brazil