Label-Free Proteomics of a Defined, Binary Co-culture Reveals Diversity of Competitive Responses Between Members of a Model Soil Microbial System.

Interactions among members of microbial consortia drive the complex dynamics in soil, gut, and biotechnology microbiomes. Proteomic analysis of defined co-cultures of well-characterized species provides valuable information about microbial interactions. We used a label-free approach to quantify the responses to co-culture of two model bacterial species relevant to soil and rhizosphere ecology, Bacillus atrophaeus and Pseudomonas putida. Experiments determined the ratio of species in co-culture that would result in the greatest number of high-confidence protein identifications for both species. The 281 and 256 proteins with significant shifts in abundance for B. atrophaeus and P. putida, respectively, indicated responses to co-culture in overall metabolism, cell motility, and response to antagonistic compounds. Proteins associated with a virulent phenotype during surface-associated growth were significantly more abundant for P. putida in co-culture. Co-culture on agar plates triggered a filamentous phenotype in P. putida and avoidance of P. putida by B. atrophaeus colonies, corroborating antagonistic interactions between these species. Additional experiments showing increased relative abundance of P. putida under conditions of iron or zinc limitation and increased relative abundance of B. atrophaeus under magnesium limitation were consistent with patterns of changes in abundance of metal-binding proteins during co-culture. These results provide details on the nature of interactions between two species with antagonistic capabilities. Significant challenges remaining for the development of proteomics as a tool in microbial ecology include accurate quantification of low-abundance peptides, especially from rare species present at low relative abundance in a consortium.

Orthotopic liver transplantation without venovenous bypass: 125 cases from a single center.

AIM: This study analyzed a 10-year single-center experience in orthotopic liver transplantation (OLT) without venovenous bypass (VVB). METHODS: We retrospectively analysed a nonrandomized series (1999-2008) of 125 adult OLT patients without VVB. RESULTS: The main causes of liver failure were viral hepatitis (n = 39), alcoholic liver disease (n = 22), and liver cancer (n = 17). One-year survival was 76.4%. The most common postoperative complications were bile duct stenosis (n = 12), postoperative bleeding (n = 8), hepatic artery thrombosis (n = 7), and primary liver failure (n = 6). Twelve patients required hemodialysis and four underwent retransplantations of the liver. Fourteen patients died before postoperative day 30(th). Univariate analysis showed significant differences between patients who did and did not survive 30 days among donor death diagnoses (P = .05), red blood cell units transfused (P = .03), aspartate aminotranferase on the first postoperative day (P = .002), ABO type (P = .04), time of orotracheal intubation (P = .001), hemodialysis (P = .001), and period of postoperative vasoactive drug use (P = .006). The total length of orotracheal tube intubation showed a significant independent association with mortality before 30 days (P < .001). CONCLUSION: OLT without VVB can be safely performed even in severe cases of chronic liver failure.

Orthotopic liver transplantation without venovenous bypass using the conventional and piggyback techniques.

INTRODUCTION: Orthotopic liver transplantation is a widely used procedure for the treatment of irreversible liver diseases for which there is no possibility of medical treatment. When this procedure is performed by the conventional technique, the retrohepatic vena cava is removed along with the native liver. The inferior vena cava (IVC) remains clamped until the revascularization of the graft, and in this period there is a reduction in the venous return, which may induce a fall by up to 50% in the cardiac output with hemodynamic instability and a fall in renal perfusion pressure. The use of a portal-femoral-axillary venovenous bypass system, in which the blood from the femoral and portal veins returns to the heart via the axillary vein propelled by a centrifugal pump, is intended to minimize the effects of the IVC clamping. In the piggyback (PB) technique, the native liver is removed and the IVC of the recipient is preserved and only partially clamped. We have employed both techniques without the use of venovenous bypass for 10 years. The objective of this study was to compare the results obtained from the use of the two techniques. PATIENTS AND METHODS: A retrospective analysis was performed of 195 patients transplanted between 1999 and 2008: 125 by the conventional technique and 70, the PB technique. The intraoperative parameters were analyzed (surgical time, ischemia time, use of blood products, and diuresis), as well as intensive care support (duration of stay in intensive care unit and use of vasoactive drugs), period of intubation, length of hospital stay, renal function, graft function, postoperative complications, retransplantation, and patient survival. RESULTS: The PB group showed a reduction in surgical time, warm ischemia time, the use of packed red blood cells concentrates, and fresh frozen plasma, as well as mortality at 30 days (P<.05). There were no differences in relation to cold ischemia time, intraoperative diuresis; length of stay and use of vasoactive drugs in the intensive care unit; the period of intubation; the duration of hospital stay; the renal function; the graft function; the need for reoperation; the incidence of sepsis, biliary complications, vascular complications; need for retransplantation; and 1-year mortality. The cumulative survival rate at 1 year was significantly better among the PB patients. CONCLUSION: Orthotopic liver transplantation can be performed without venovenous bypass with good results, using either the conventional technique or the PB technique. Provided that there is no technical contraindication and a long ischemia period is not foreseen, the PB technique should be the technique of choice.

Survival on waiting list for liver transplantation before and after introduction of the model for end-stage liver disease score.

BACKGROUND: Since July 2006, the Model for End-stage Liver Disease (MELD) score has served as the national basis for allocation of donor livers for transplantation in Brazil. Patients with higher MELD scores receive greater priority for allocation regardless of the time on the waiting list. PURPOSE: To investigate the impact of MELD score implementation on the survival of waiting list patients. METHODS: A retrospective study of patients registered at the national Organ Procurement Organization (OPO) for the liver transplantation waiting list between January 2004 and June 2006 (pre-MELD) and between July 2006 and December 2008 (post-MELD). RESULTS: We included listed patients awaiting liver transplantation in the pre-MELD era (n = 250, 48.4%) and in the post-MELD era (n = 266, 51.6%). The times awaiting transplant prior to and after the MELD system were 487.2 +/- 384.8 days and 183.9 +/- 157.2 days, respectively. Prior to the MELD score, waiting list survivals were greater when compared to rates in the current system. Early posttransplant patient survival rates were significantly reduced in the post-MELD era (83.4%) compared to the period before MELD implementation (93.2%). CONCLUSIONS: MELD score provides a transparent, objective system to drive allocation policy; however, it presents several important limitations. Constant need of changes and reevaluation are needed as an evolutionary process. Future changes in the present system may be addressed by adjusting the MELD system.

Reconstruction of venous outflow of domino liver graft using deceased donor iliac/cava veins.

The double piggyback technique has been proposed for domino liver transplantation. To make this possible, it is necessary to reconstruct the venous outflow of the domino liver graft on the back table. The authors describe the technical details of this procedure in three consecutive cases. A deceased donor cava-iliac bifurcation segment was used. The iliac veins were anastomosed to the ostia of the right and middle-left hepatic veins, and the graft cava vein was anastomosed to the ostium of the three hepatic veins of the recipient. In all cases anatomic compatibility was observed; the outcome of the patients was satisfactory.

Transplante de fígado em dominó com técnica de duplo piggy back / Domino liver transplantation with the double pigg back technique

Os autores apresentam um caso de transplante hepático em dominó realizado com nova técnica cirúrgica. A paciente, portadora de Polineuropatia Amiloidótica Familiar Tipo I (PAF) foi submetida a hepatectomia pela técnica piggback, recebendo fígado de cadáver. O segundo paciente com cirrose vírus B e carcinoma hepatocelular, receptor dominó, também foi submetido a hepatectomia com preservação da veia cava recebendo enxerto após reconstruções vasculares (venosa e arterial). Ambos tiveram alta sem intercorrências. Os relatos demonstram que a modalidade de transplante hepático dominó é factível com a técnica de duplo piggback, prescindindo do desvio porto-cava-axilar

Pituitary apoplexy. Clinical course, endocrine evaluations and treatment analysis.

BACKGROUND: The purpose was to analyze clinical manifestations, hormonal changes, diagnosis difficulties and treatment of pituitary apoplexy (PA). EXPERIMENTAL DESIGN: A retrospective study of clinical records from patients with pituitary adenomas admitted from January 1980 to June 1996; the purpose was to identify the patients with clinical evidence compatible with PA. SETTING: Neurosurgery unit of an institutional hospital. PATIENTS: Sixteen (12.8%) of 125 patients with pituitary adenomas were analyzed because they had pituitary apoplexy. INTERVENTIONS: Surgical treatment by the trans-sphenoidal or transcranial route or both routes; dexamethasone (DXM) treatment with 16 mg/day i.v. MEASURES: Hormone assays were performed either by radioimmunoassay or by chemical luminescence. RESULTS: Tumors were nonfunctioning in nine patients and functioning in seven. TSH and prolactin basal serum levels were impaired in 55.5% and 10%, respectively; after exogenous TRH 80% of the patients did not show stimulation of TSH and prolactin secretions. LH and FSH levels were low in 63.6% and 54.6% of the patients, respectively; gonadotrophin-releasing hormone (GnRH) testing was abnormal in 75% of the patients evaluated. Cortisol levels were low in 50% of the patients. After insulin-induced hypoglycemia, cortisol and GH failed to rise in 25% and 40% of cases, respectively. Ten patients were submitted to surgical treatment, but none during PA. The average time from the onset of apoplectic symptoms and surgery was 70+/-50 days. Only one patient died two months after surgery. Five patients were treated with dexamethasone (DXM) during the apoplectic symptoms: three patients died; one patient had good quality of life; the other patient was treated initially with DXM with improvement of vision, but after surgery he developed panhypopituirarism. Two other patients did not receive specific treatment for PA. CONCLUSIONS: PA is not a rare pituitary adenoma complication and its prognosis may be poor; baseline hormone levels showed a wide range of abnormalities of pituitary function; surgical treatment was required in the majority of patients and the prognosis was relatively good; on the contrary, the treatment with DXM only had high levels of mortality.

Advanced megaesophagus with recurrent dysphagia following initial surgical treatment.

The results of the surgical treatment of twenty patients with advanced megaesophagus who had undergone previous treatment are presented. After the previous operation, the asymptomatic period was less than five years in mot cases; symptoms included dysphagia (100%), regurgitation (65%), heartburn (50%), pain (45%), excess saliva (20%) and palpitations (10%). The definitive treatment was cervico-abdominal esophagectomy (45%), distal esophagectomy (20%), Thal-Hatafuku's operation (15%) and miscellaneous (15%). In our experience, the best procedures are cervico-abdominal esophagectomy and Thal-Hatafuku's operation.

Proximal gastric vagotomy in stenosed or perforated duodenal ulcer.

Thirty-five patients suffering from stenosed or perforated duodenal ulcer, who were submitted to surgical treatment in the form of proximal gastric vagotomy, are presented. There were no deaths in the series and the results were considered to be very good in the cases of perforation, whereas a high incidence of poor results (27 per cent) occurred in the cases of stenosis submitted to a proximal gastric vagotomy combined with digital dilatation of the pylorus.

A vagotomia gastrica proximal no tratamento cirurgico da ulcera duodenal estenosada e perfurada. / Proximal gastric vagotomy in the surgical treatment of stenosing and perforating duodenal ulcer

Os autores apresentam um material de 35 pacientes portadores de ulcera duodenal estenosada e perfurada, submetidos a tratamento cirurgico utilizado a vagotomia gastrica proximal. Nao houve mortalidade na serie e os resultados foram considerados muito bons nos casos de perfuracao, ocorrendo elevado indice de maus resultados (27,27) nos casos de estenose submetidos a uma vagotomia gastrica proximal associada a dilatacao digital do pilor

Tratamento cirurgico do megaesofago chagasico. Avaliacao de 60 casos. / Surgical treatment of chagasic megaesophagus

Os autores estudaram retrospectivamente o resultado do tratamento cirurgico de 60 pacientes portadores de megaesofago, sendo 22 submetidos a operacao de Heller associada a um procedimento antirefluxo, 22 a operacao de Thal Hatafuku e 16 a esofogectomia cervico-abdominal (Op. Eugenio Ferreira).O seguimento pos-operatorio global foi de 73,33% e ocorreram dois casos de obito entre os submetidos a operacao de Thal Hatafuku (4,54%). Os resultados considerados bons e regulares ocorreram em 59,11% dos casos submetidos a operacao de Heller associada a um procedimento antirefluxo, 40,92% apos a operacao de Thal Hatafuku e 87,50% apos a operacao de Eugenio Ferreira