Embriogênese somática do mamoeiro e estabelecimento de um protocolo de seleção com Glufosinato de Amônio (GA) visando à geração de plantas geneticamente modificadas.

O Brasil é o principal produtor mundial de mamão, sendo responsável por 10% da área plantada e 25% da produção mundial em 2006, o que mostra uma alta produtividade da cultura no país. O desenvolvimento da cultura do mamoeiro tem sido limitado por diversos fatores como os relacionados à sua natureza dióica, à heterozigose, à suscetibilidade a doenças e à falta de métodos comerciais de multiplicação vegetativa. A embriogênese somática tem sido bem estudada em vários cultivares comerciais de mamoeiro. Esta técnica é, portanto, o meio pelo qual células somáticas se desenvolvem em estruturas que se assemelham aos embriões zigóticos (isto é, bipolar e sem conexão vascular ao tecido parental) com uma série de estádios embriológicos característicos, sem fusão de gametas. O presente trabalho teve como objetivo induzir e desenvolver calos e embriões somáticos da variedade de mamoeiro Sunrise a partir de sementes oriundas de frutos imaturos de mamão presentes no BAG de mamão (Banco Ativo de Germoplasma) situado na Embrapa Mandioca e Fruticultura. Os frutos imaturos com aproximadamente 90 a 120 dias foram coletados no BAG, lavados com água corrente e hipoclorito 2%, e posteriormente, encaminhados para câmara de fluxo laminar, onde foi realizado o processo de desinfestação em condições assépticas. O meio de cultura foi composto de 1/2MS suplementado com 10mg/L de 2,4-D (ácido diclorofenoxiacético), 60g/L de sacarose, 10ml/L de vitamina de mamão (L-Glutamina, myo-inositol, thiamina, glycine, ácido nicotínico e piridoxina), 8g/L de ágar e pH 5,8. Para o estabelecimento da curva de seleção com calos embriogênicos de mamão utilizando como agente seletivo glufosinato de amônio. As concentrações usadas foram 0; 1; 2,5; 5; 10 e 15 mg/L. A formação de calos contendo embriões somáticos primários iniciou-se três a quatro semanas após a inoculação in vitro dos explantes, contudo pode-se observar um maior número de explantes em estádios mais tardios (torpedo e cotiledonar) após cinco semanas de cultivo. Para o experimento da curva de seleção com o herbicida glufosinato de amônio, foi possível observar inibição no crescimento e desenvolvimento de embriões somáticos nas concentrações de 5, 10 e 15 mg/L. Nossos resultados sugerem que o protocolo de indução de embriogênese somática de mamoeiro testado foi reproduzível, e apresentou uma alta frequência na indução de embriões somáticos secundários. O sistema Bar/Glufosinato de amônio (GA) para a seleção de embriões transgênicos de mamoeiro, deve ser realizado em concentração igual ou superior a 5 mg/L de GA.PDF. T27

Recent Innovations & Daily Problems. A new prosthesis in inguinal hernia repair:preliminary results of a pilot study.

Introduction: Elective surgery for inguinal hernia is affected by very low mortality « 1 per 10000 operation); in contrast, when surgery is carried out for complicated inguinal hernia, risks of postoperative complication are higher. TAPP is a world-wide accepted surgical practice in the treatment of elective bilateral or recurrent inguinal hernia, above all in young patients. Few exploratory studies were published on laparoscopic approach in the treatment of urgent complicated inguinal hernia. Aim of this study was to analyze feasibility (operative time, conversion rate), safety (postoperative morbidity, length of hospital stay) and quality of life (acute and chronic pain, return to work) of trans-abdominal pre-peritoneal laparoscopic hernia repair in acute incarcerated inguinal hernia. Rationale of laparoscopic trans-abdominal approach is the easier hernia reduction under vision and a better exploration of the abdominal cavity. Methods: from September 2012 to September 2013, 15 consecutive patients admitted in emergency at the Division of General Surgery of University "Sapienza", Polo Pontino, for acute incarcerated inguinal hernia were submitted to TAPP using 3 trocars (1 of 10 mm and 2 of 5mm) and polyester prosthesis fixed by fibrin glue. Exclusion criteria for laparoscopic approach were age III, previous abdominal surgery, signs of strangulated hernia. All of them were evaluated for operative time, conversion rate, postoperative morbidity, organ resection or other surgery required. All patients were scored for pain by Visual Analogic Scale (VAS) during postoperative in hospital stay at 7 days, 1,6 and 12 months after surgery. Results: median follow-up was 16 months and 12 as minimum. In all cases reduction of hernia was always possible and none conversion to open surgery was recorded, median operative time was 89 minutes (55-137 as range), omental resection was carried out in one patient (6,6%), no other organ resections needed, whereas contralateral hernia was diagnosed and repaired at the same time in 4 patients (26,6%). No major complications were observed, median blood loss was 100 ml, minor morbidity was contained to 18% represented by fever and wound infection of surgical umbilical scar. Median in hospital stay was 1,5 days with 1-5 days as range. Postoperative median acute pain, measured by visual analogic scale (VAS), was 2 (range:0-4), none patient referred any pain during follow-up. Median time of return to work was 6,5 days, ranged between 3 to 15 days. Patients' compliance to treatment and to follow-up was complete as well their satisfaction. Conclusions: In centres skilled for laparoscopy in emergency, TAPP could be considered a feasible and safe technique. In well-selected patients (especially if emolled in controlled clinical trial) TAPP could represent an alternative surgical approach for complicated incarcerated inguinal hernia to conventional open surgery even in urgency. The main advantages of laparoscopic approach are the ability to perform surgical hernia reduction under vision, a better exploration and evaluation of abdominal cavity and diagnosis and treatment of eventual contralateral defect of wall, otherwise often missed. Finally, the good control of acute and chronic pain, faster return to normal activity and work, better aesthetic results contributed to total satisfaction and compliance of the patients

Rationale, study design, and analysis plan of the Alveolar Recruitment for ARDS Trial (ART): study protocol for a randomized controlled trial

BACKGROUND: Acute respiratory distress syndrome (ARDS) is associated with high in-hospital mortality. Alveolar recruitment followed by ventilation at optimal titrated PEEP may reduce ventilator-induced lung injury and improve oxygenation in patients with ARDS, but the effects on mortality and other clinical outcomes remain unknown. This article reports the rationale, study design, and analysis plan of the Alveolar Recruitment for ARDS Trial (ART). METHODS/DESIGN: ART is a pragmatic, multicenter, randomized (concealed), controlled trial, which aims to determine if maximum stepwise alveolar recruitment associated with PEEP titration is able to increase 28-day survival in patients with ARDS compared to conventional treatment (ARDSNet strategy). We will enroll adult patients with ARDS of less than 72 h duration. The intervention group will receive an alveolar recruitment maneuver, with stepwise increases of PEEP achieving 45 cmH2O and peak pressure of 60 cmH2O, followed by ventilation with optimal PEEP titrated according to the static compliance of the respiratory system. In the control group, mechanical ventilation will follow a conventional protocol (ARDSNet). In both groups, we will use controlled volume mode with low tidal volumes (4 to 6 mL/kg of predicted body weight) and targeting plateau pressure 6430 cmH2O. The primary outcome is 28-day survival, and the secondary outcomes are: length of ICU stay; length of hospital stay; pneumothorax requiring chest tube during first 7 days; barotrauma during first 7 days; mechanical ventilation-free days from days 1 to 28; ICU, in-hospital, and 6-month survival. ART is an event-guided trial planned to last until 520 events (deaths within 28 days) are observed. These events allow detection of a hazard ratio of 0.75, with 90% power and two-tailed type I error of 5%. All analysis will follow the intention-to-treat principle. DISCUSSION: If the ART strategy with maximum recruitment and PEEP titration improves 28-day survival, this will represent a notable advance to the care of ARDS patients. Conversely, if the ART strategy is similar or inferior to the current evidence-based strategy (ARDSNet), this should also change current practice as many institutions routinely employ recruitment maneuvers and set PEEP levels according to some titration metho