The Valgent4 protocol:Robust analytical and clinical validation of 11 HPV assays with genotyping on cervical samples collected in SurePath medium

BACKGROUND: The VALidation of HPV GENoyping Tests (VALGENT) is an international initiative designed to validate HPV assays with genotyping capability. The VALGENT4 protocol differs from previous VALGENT installments as the sample collection medium is SurePath, and exclusively includes samples from women ≥30 years of age which is concordant with the majority of HPV primary screening guidelines. Here we present the protocol for the fourth installment of the VALGENT framework. OBJECTIVES: In VALGENT4 11 HPV assays will be evaluated using two comparator assays based on PCR with the GP5+/6+ primers. STUDY DESIGN: Overall, the VALGENT4 panel consists of 1,297 routine samples comprised of 998 unselected, consecutive samples, of which 51 samples had abnormal cytology with 13 women diagnosed with ≥CIN2, and 299 consecutive samples enriched for ≥ASCUS cytology (100 ASCUS, 100 LSIL, 99 HSIL) with 106 ≥CIN2 upon follow up. Manipulated and DNA extracted panel samples were characterized with respect to human beta globin (HBB) and overall DNA content and composition to quality assess the panel prior to distribution to the collaborating sites. RESULT: The relative cellularity (mean CT value of HBB from the Onclarity assay) on the 1,297 LBC samples (CT=24.8) was compared with 293 un-manipulated routine cytology screening samples (CT=23.8). Furthermore, the DNA extracted panel samples was characterized using the Exome iPLEX pro assay, which reports amplifiable copies on individual samples as well as copies of five different base pair lengths. Here the data showed a slightly lower number of amplifiable DNA copies (ratio: 0.7, p=<0.01)) in the VALGENT4 panel samples compared to routine extracted cervical DNA samples CONCLUSION: The present manuscript details the manipulation, processing and quality assessment of samples used in VALGENT-4. This methodological document may be of value for future international projects of HPV test validation

Quesungual slash and mulch agroforestry system (QSMAS): Improving crop water productivity, food security and resource quality in the sub-humid tropics

The knowledge and principles generated by CPWF-PN15 confirm that QSMAS can be a model production system for implementing conservation agriculture to achieve food security and sustainable development in drought-prone areas of hillsides in the sub-humid tropics, while providing ecosystem services in the face of land degradation and climate change. As an adoptable option to replace the slash and burn traditional system, QSMAS can improve smallholder livelihoods through eco-efficient use and conservation of natural resources. Participatory validation activities suggest that the conservation agriculture principles embedded in QSMAS can be readily accepted by resource- poor farmers and local authorities in similar agroecosystems

Canagliflozin and renal outcomes in type 2 diabetes and nephropathy

BACKGROUND Type 2 diabetes mellitus is the leading cause of kidney failure worldwide, but few effective long-term treatments are available. In cardiovascular trials of inhibitors of sodium–glucose cotransporter 2 (SGLT2), exploratory results have suggested that such drugs may improve renal outcomes in patients with type 2 diabetes. METHODS In this double-blind, randomized trial, we assigned patients with type 2 diabetes and albuminuric chronic kidney disease to receive canagliflozin, an oral SGLT2 inhibitor, at a dose of 100 mg daily or placebo. All the patients had an estimated glomerular filtration rate (GFR) of 30 to &lt;90 ml per minute per 1.73 m2 of body-surface area and albuminuria (ratio of albumin [mg] to creatinine [g], &gt;300 to 5000) and were treated with renin–angiotensin system blockade. The primary outcome was a composite of end-stage kidney disease (dialysis, transplantation, or a sustained estimated GFR of &lt;15 ml per minute per 1.73 m2), a doubling of the serum creatinine level, or death from renal or cardiovascular causes. Prespecified secondary outcomes were tested hierarchically. RESULTS The trial was stopped early after a planned interim analysis on the recommendation of the data and safety monitoring committee. At that time, 4401 patients had undergone randomization, with a median follow-up of 2.62 years. The relative risk of the primary outcome was 30% lower in the canagliflozin group than in the placebo group, with event rates of 43.2 and 61.2 per 1000 patient-years, respectively (hazard ratio, 0.70; 95% confidence interval [CI], 0.59 to 0.82; P=0.00001). The relative risk of the renal-specific composite of end-stage kidney disease, a doubling of the creatinine level, or death from renal causes was lower by 34% (hazard ratio, 0.66; 95% CI, 0.53 to 0.81; P&lt;0.001), and the relative risk of end-stage kidney disease was lower by 32% (hazard ratio, 0.68; 95% CI, 0.54 to 0.86; P=0.002). The canagliflozin group also had a lower risk of cardiovascular death, myocardial infarction, or stroke (hazard ratio, 0.80; 95% CI, 0.67 to 0.95; P=0.01) and hospitalization for heart failure (hazard ratio, 0.61; 95% CI, 0.47 to 0.80; P&lt;0.001). There were no significant differences in rates of amputation or fracture. CONCLUSIONS In patients with type 2 diabetes and kidney disease, the risk of kidney failure and cardiovascular events was lower in the canagliflozin group than in the placebo group at a median follow-up of 2.62 years

Celecoxib induces anoikis in human colon carcinoma cells associated with the deregulation of focal adhesions and nuclear translocation of p130Cas

Farre, Maria Lourdes Vallve “Documento produzido em parceria ou por autor vinculado à Fiocruz, mas não consta à informação no documento”.Submitted by Ana Maria Fiscina Sampaio ([email protected]) on 2017-05-29T13:59:07Z No. of bitstreams: 1 Casanova I Calecoxib induces anoikis in human.....pdf: 905227 bytes, checksum: 0134e8f1a744a3d891d16c446b731bb7 (MD5)Approved for entry into archive by Ana Maria Fiscina Sampaio ([email protected]) on 2017-05-29T14:16:25Z (GMT) No. of bitstreams: 1 Casanova I Calecoxib induces anoikis in human.....pdf: 905227 bytes, checksum: 0134e8f1a744a3d891d16c446b731bb7 (MD5)Made available in DSpace on 2017-05-29T14:16:25Z (GMT). No. of bitstreams: 1 Casanova I Calecoxib induces anoikis in human.....pdf: 905227 bytes, checksum: 0134e8f1a744a3d891d16c446b731bb7 (MD5) Previous issue date: 2006Grant sponsor: The Spanish Government; Grant numbers: SAF03/07437, FIS 01/0853, FISC03/10; Grant sponsor: Spanish National Health System; Grant numbers: FIS 98/3197, FIS 01/3085Hospital de la Santa Creu i Sant Pau. Laboratori d’Investigaci o Gastrointestinal de l’Institut de Recerca. Barcelona, SpainHospital de la Santa Creu i Sant Pau. Laboratori d’Investigaci o Gastrointestinal de l’Institut de Recerca. Barcelona, SpainHospital de la Santa Creu i Sant Pau. Laboratori d’Investigaci o Gastrointestinal de l’Institut de Recerca. Barcelona, SpainHospital de la Santa Creu i Sant Pau. Laboratori d’Investigaci o Gastrointestinal de l’Institut de Recerca. Barcelona, SpainHospital de la Santa Creu i Sant Pau. Laboratori d’Investigaci o Gastrointestinal de l’Institut de Recerca. Barcelona, SpainHospital de la Santa Creu i Sant Pau. Laboratori d’Investigaci o Gastrointestinal de l’Institut de Recerca. Barcelona, SpainHospital de la Santa Creu i Sant Pau. Deparment of Pathology. Barcelona, SpainHospital de la Santa Creu i Sant Pau. Deparment of Medical Oncology. Barcelona, SpainHospital de la Santa Creu i Sant Pau. Deparment of Surgery. Barcelona, SpainHospital de la Santa Creu i Sant Pau. Laboratori d’Investigaci o Gastrointestinal de l’Institut de Recerca. Barcelona, SpainCelecoxib, a selective cyclooxygenase-2 (COX-2) inhibitor, is effective as chemopreventive against colon cancer and it is the only nonsteoroidal antiinflammatory drug approved by the FDA for adjuvant therapy in patients with familial adenomatous polyposis. It is also being evaluated, within Phase II and III clinical trials, in combination with standard chemotherapy to treat sporadic colorectal cancer. Nevertheless, its antitumor mechanism of action is still not fully understood. In this study, we have evaluated the in vitro growth inhibitory effect of celecoxib in colon carcinoma cells and analyzed its mechanism of action. We report that the deregulation of the focal adhesion assembly protein Crk-associated substrate 130 kDa (p130Cas) by celecoxib plays a relevant role in the cytotoxic effect of this drug. Thus, celecoxib induces the proteolysis of p130Cas and the nuclear translocation of the 31 kDa generated fragment leading to apoptosis. Furthermore, overexpression of wild-type p130Cas reverts, in part, the growth inhibitory effect of celecoxib. In contrast, FAK and AKT do not appear to be involved in this activity. Our data suggest, for the first time, that the antitumor mechanism of action of celecoxib includes the induction of anoikis, an effect that is not related to COX-2 inhibition. Besides providing new insights into the antitumor effect of celecoxib, this novel mechanism of action holds potential relevance in drug development. Indeed, our results open the possibility to develop new celecoxib derivatives that induce anoikis without COX-2 inhibition so as to avoid the cardiovascular toxicity recently described for the COX-2 inhibitors