Flávio Henrique Ferreira Galvão

Intrahepatic Glissonian approach for laparoscopic right segmental liver resections.

BACKGROUND Experience with laparoscopic procedures and recent advances in laparoscopic devices have created an evolving interest in the application of these techniques to liver resection. However, laparoscopic liver resection has not been widely developed and anatomical segmental liver resection is not currently performed due to difficulty to control segmental Glissonean pedicles laparoscopically. METHODS Seven consecutive patients underwent laparoscopic liver resection using an intrahepatic Glissonian approach from April 2007 to September 2007. Three patients underwent laparoscopic bisegmentectomy 6-7 and 4 patients underwent laparoscopic right hemihepatectomy. RESULTS Blood transfusion was required in 1 patient. Mean operation time was 460 minutes (range 300-630 minutes). The median hospital stay was 5 days (range 3-8 days). One patient developed bile leakage that was treated conservatively. No patient had postoperative signs of liver failure. No postoperative mortality was observed. CONCLUSIONS The main advantage over other techniques is the possibility to gain a rapid and precise access to the right posterior and anterior sheaths facilitating right hemihepatectomy, and right anterior and posterior sectionectomies. We believe that the described technique facilitates laparoscopic liver resection by reducing the technical difficulties in pedicle control and may increase the development of segment-based laparoscopic liver resections.

Marginal grafts increase early mortality in liver transplantation

CONTEXT AND OBJECTIVE Expanded donor criteria (marginal) grafts are an important solution for organ shortage. Nevertheless, they raise an ethical dilemma because they may increase the risk of transplant failure. This study compares the outcomes from marginal and non-marginal graft transplantation in 103 cases of liver transplantation due to chronic hepatic failure. DESIGN AND SETTING One hundred and three consecutive liver transplantations to treat chronic liver disease performed in the Liver Transplantation Service of Hospital das Clínicas da Faculdade de Medicina da Universidade de São Paulo between January 2001 and March 2006 were retrospectively analyzed. METHODS We estimated graft quality according to a validated scoring system. We assessed the pre-transplantation liver disease category using the Model for End-Stage Liver Disease (MELD), as low MELD (< 20) or high MELD (>or= 20). The parameters for marginal and non-marginal graft comparison were the one-week, one-month and one-year recipient survival rates, serum liver enzyme peak, post-transplantation hospital stay and incidence of surgical complications and retransplantation. The significance level was 0.05. RESULTS There were no differences between the groups regarding post-transplantation hospital stay, serum liver enzyme levels and surgical complications. In contrast, marginal grafts decreased overall recipient survival one month after transplantation. Furthermore, low-MELD recipients of non-marginal grafts showed better one-week and one-month survival than did high-MELD recipients of marginal livers. After the first month, patient survival was comparable in all groups up to one year. CONCLUSION The use of marginal graft increases early mortality in liver transplantation, particularly among high-MELD recipients.

Conhecimento e opinião de estudantes de medicina sobre doação e transplante de órgãos

We analyzed the opinion and understanding of medical students about organ donation and transplantation. METHODS: 347 students voluntarily completed a questionnaire with 17 queries concerning organ donation and transplantation. They were analyzed to identify general tendencies and divided into five groups, according to their year of study (first through sixth year), to assess differences among the years. Students of the fifth and sixth years were placed in the same group. RESULTS:were analyzed by the Chi-square test. RESULTS: The intention to become a post mortem or living donor was of 89% and 90% respectively; however, only 62% were aware of living donation risks. 70% of the 347 students admitted regular or little knowledge of the subject, 90.2% considered organ transplantation an important issue for a medical graduation program, 76.9% considered informed/expressed consent the best organ donation criterion and 64.3% of them chose severity of patient disease as the best allocation condition. As students progressed in their studies their understanding about transplantation improved. Students of the fourth, fifth and sixth year manifested a negative attitude about organ donation to alcohol addicts, non donors, drug users, law offenders and foreigners. CONCLUSION: This data show the great interest and positive attitude of medical students toward organ donation and transplantation, despite the fact that most of them admitted having insufficient knowledge on the subject. A negative attitude by students of the fourth, fifth and sixth year on organ donation to alcohol addicts, non donors, drug users, law offenders and foreigners was also observed.

Glutathione ethyl ester supplementation during pancreatic islet isolation improves viability and transplant outcomes in a murine marginal islet mass model.

Background The success of pancreatic islet transplantation still faces many challenges, mainly related to cell damage during islet isolation and early post-transplant. The increased generation of reactive oxygen species (ROS) during islet isolation and the consumption of antioxidant defenses appear to be an important pathway related to islet damage. Methodology/Principal Findings In the present study we evaluated whether supplementation of glutathione-ethyl-ester (GEE) during islet isolation could improve islet viability and transplant outcomes in a murine marginal islet mass model. We also cultured human islets for 24 hours in standard CMRL media with or without GEE supplementation. Supplementation of GEE decreased the content of ROS in isolated islets, leading to a decrease in apoptosis and maintenance of islet viability. A higher percentage of mice transplanted with a marginal mass of GEE treated islets became euglycemic after transplant. The supplementation of 20 mM GEE in cultured human islets significantly reduced the apoptosis rate in comparison to untreated islets. Conclusions/Significance GEE supplementation was able to decrease the apoptosis rate and intracellular content of ROS in isolated islets and might be considered a potential intervention to improve islet viability during the isolation process and maintenance in culture before islet transplantation.

Intestinal transplantation including anorectal segment in the rat

Transplantation of small bowel and colon has been recently advocated, aiming to prevent dehydration, reduce stoma output, and promote earlier post-transplant weaning from parental nutrition. Some patients needing intestinal transplantation may also have anorectal dysfunction or a permanent colostomy, which impairs quality of life. The inclusion of the anorectal segment in the intestinal graft may be an attractive improvement for such patients. In this letter, we describe a model of en bloc intestinal transplantation in the rat, including jejunum, ileum, cecum, entire colon, rectum, and the anus. In donor operation, we performed a combined perianal and midline abdominal incision (Fig. 1A). The completely dissected anorectal segment was mobilized to inside the abdomen through the perineum, sectioning rectal vessels and pudendal nerves and preserving the inferior mesenteric artery and vein. The superior mesenteric and portal veins were separated from the pancreas by division of duodenal, splenic, and left gastric veins. The abdominal aorta was dissected from the iliac bifurcation up to the diaphragm by dividing lumbar and renal arteries and the celiac trunk (Fig. 1B). After heparinization, we tied the aorta distally to the inferior mesenteric artery and cut it near the diaphragm and after the tie to perform a long aortomesenteric conduit including the superior and inferior mesenteric arteries. The portal vein was dissected and cut near the hepatic hilum, the proximal jejunum was sectioned, and the graft was removed and placed in cold preservation solution. During back table procedures, we placed a cuff in portal vein, as previously described (Fig. 1C). In the recipient, we performed the same combined incision and anorectal mobilization. The infrarenal abdominal aorta was cross-clamped, and a continuous end-to-side hand-sewn microanastomosis was performed between recipient’s aorta and donor’s aortomesenteric conduit. Below the second jejunal branch, we dissected approximately 1 cm of the recipient superior mesenteric vein, which was clamped and sectioned. The donor portal cuff was introduced in the recipient’s superior mesenteric vein, and an encircling ligature was performed to fix the cuff and complete the anastomosis. Subsequently, we tied and divided the superior and inferior mesenteric arteries and removed en bloc the native mesentery, jejunum, ileum, cecum, entire colon, rectum, and anus. After clamps removal, immediate pulsation of the aortomesenteric conduit and flow throughout the portal vein were observed (Fig. 1D). To restore the intestinal tract, recipient’s and donor’s jejunum were anastomosed, and the anorectal segment was replaced in its ortothopic position and fixed by stitches between donor’s skin surrounding the anus and recipient’s perineal skin. Total operating time was about 3 hours and total ischemia time was about 60 minutes. This intestinal transplantation modification of our previously described models preserves bowel intrinsic innervation, ileocecal valve, and anal sphincter, which may improve graft physiology. It also allows studies evaluating the regeneration of intestinal and anorectal innervation by anorectal electromyography and manometry. Furthermore, it maintains physiological portal drainage and may be useful for metabolic and Presented at the 10th Congress of the International Society for Experimental Microsurgery (ISEM), São Paulo, Brazil, October 29th–November 1st, 2010. *Correspondence to: Flávio Henrique Ferreira Galvão, Av. Dr. Arnaldo, 455Cerqueira Cesar, Room 3206, São Paulo, SP, Brazil. E-mail: fgalvao@usp.br Received 30 April 2011; Accepted 9 August 2011 Published online 17 October 2011 in Wiley Online Library (wileyonlinelibrary. com). DOI 10.1002/micr.20958

Intestinal Translocation of Clinical Isolates of Vancomycin-Resistant Enterococcus faecalis and ESBL-Producing Escherichia coli in a Rat Model of Bacterial Colonization and Liver Ischemia/Reperfusion Injury

The objectives of this study were to develop a rat model of gastrointestinal colonization with vancomycin-resistant Enterococcus faecalis (VRE) and extended-spectrum beta-lactamase (ESBL)-producing E. coli and to evaluate intestinal translocation to blood and tissues after total and partial hepatic ischemia. Methods - We developed a model of rat colonization with VRE and ESBL-E coli. Then we studied four groups of colonized rats: Group I (with hepatic pedicle occlusion causing complete liver ischemia and intestinal stasis); Group II (with partial liver ischemia without intestinal stasis); Group III (surgical manipulation without hepatic ischemia or intestinal stasis); Group IV (anesthetized without surgical manipulation). After sacrifice, portal and systemic blood, large intestine, small intestine, spleen, liver, lungs, and cervical and mesenteric lymph nodes were cultured. Endotoxin concentrations in portal and systemic blood were determined. Results – The best inocula were: VRE: 2.4×1010 cfu and ESBL-E. coli: 1.12×1010 cfu. The best results occurred 24 hours after inoculation and antibiotic doses of 750 µg/mL of water for vancomycin and 2.1 mg/mL for ceftriaxone. There was a significantly higher proportion of positive cultures for ESBL-E. coli in the lungs in Groups I, II and III when compared with Group IV (67%; 60%; 75% and 13%, respectively; p:0.04). VRE growth was more frequent in mesenteric lymph nodes for Groups I (67%) and III (38%) than for Groups II (13%) and IV (none) (p:0.002). LPS was significantly higher in systemic blood of Group I (9.761±13.804 EU/mL−p:0.01). No differences for endotoxin occurred in portal blood. Conclusion –We developed a model of rats colonized with resistant bacteria useful to study intestinal translocation. Translocation occurred in surgical procedures with and without hepatic ischemia-reperfusion and probably occurred via the bloodstream. Translocation was probably lymphatic in the ischemia-reperfusion groups. Systemic blood endotoxin levels were higher in the group with complete hepatic ischemia.

Experimental multivisceral xenotransplantation.

Abstract:  Background:  Organ shortage impairs the proposition of multivisceral transplantation to treat multiple organ failure. Interspecies (xeno) transplantation is a valid solution for organ shortage; however, suitable models of this advance are lacking. We describe an effective model of multivisceral xenotransplantation to study hyperacute rejection.

Study of morbidity in orthotopic small intestine transplantation with Wistar rats: experimental study

BACKGROUND Transplantation of the small intestine is a surgical procedure currently under investigation for its possible application in the treatment of patients with short bowel syndrome, aiming at the reintroduction of an oral diet. AIM To define the morbidity and mortality of intestinal transplantation in small animals using microsurgery. Intra and postoperative morbidity and mortality were studied in Wistar rats submitted to orthotopic intestinal allotransplantation. MATERIAL AND METHOD The animals were divided into three groups: group A (37 donor animals), group B (37 recipient animals), and group C (10 control animals). Group B was divided into three subgroups according to survival time. Subgroup TI consisted of animals that died during surgery or due to causes directly related to surgical intervention, subgroup T2 consisted of animals that died between the 4th and 29th postoperative day, and subgroup T3 consisted of animals that survived after 30 days. Transplanted animals were evaluated in terms of surgical technique used (vascular and intestinal anastomosis), graft quality, surgical time, and clinical parameters. The animals that died by the 29th postoperative day were submitted to autopsy and the remaining ones were sacrificed after 30 days. RESULT There was a high rate of complication of a surgical nature. Early mortality rate, i.e., mortality up to the third postoperative day, was 54% with vascular anastomosis being the major cause of death. Surgical time was evaluated in a restricted and homogeneous group and showed a strong prognostic value in terms of successful transplantation. Clinical parameters such as weight loss, reduction of ingestion, reduction of motor activity and diarrhea were directly correlated with acute rejection. CONCLUSION The experimented intestinal transplant is a procedure companied by considerable morbidity and mortality due to surgical complications in postoperative period, vascular anastomosis and total surgical time.

S-Nitroso-N-Acetylcysteine Ameliorates Ischemia-Reperfusion Injury In The Steatotic Liver

BACKGROUND: Steatosis is currently the most common chronic liver disease and it can aggravate ischemia-reperfusion (IR) lesions. We hypothesized that S-nitroso-N-acetylcysteine (SNAC), an NO donor component, can ameliorate cell damage from IR injury. In this paper, we report the effect of SNAC on liver IR in rats with normal livers compared to those with steatotic livers. METHODS: Thirty-four rats were divided into five groups: I (n=8), IR in normal liver; II (n=8), IR in normal liver with SNAC; III (n=9), IR in steatotic liver; IV (n=9), IR in steatotic liver with SNAC; and V (n=10), SHAN. Liver steatosis was achieved by administration of a protein-free diet. A SNAC solution was infused intraperitoneally for one hour, beginning 30 min. after partial (70%) liver ischemia. The volume of solution infused was 1 ml/100 g body weight. The animals were sacrificed four hours after reperfusion, and the liver and lung were removed for analysis. We assessed hepatic histology, mitochondrial respiration, oxidative stress (MDA), and pulmonary myeloperoxidase. RESULTS: All groups showed significant alterations compared with the group that received SHAN. The results from the steatotic SNAC group revealed a significant improvement in liver mitochondrial respiration and oxidative stress compared to the steatotic group without SNAC. No difference in myeloperoxidase was observed. Histological analysis revealed no difference between the non-steatotic groups. However, the SNAC groups showed less intraparenchymal hemorrhage than groups without SNAC (p=0.02). CONCLUSION: This study suggests that SNAC effectively protects against IR injury in the steatotic liver but not in the normal liver.

The Impact of the Introduction of MELD on the Dynamics of the Liver Transplantation Waiting List in São Paulo, Brazil

Until July 15, 2006, the time on the waiting list was the main criterion for allocating deceased donor livers in the state of São Paulo, Brazil. After this date, MELD has been the basis for the allocation of deceased donor livers for adult transplantation. Our aim was to compare the waitlist dynamics before MELD (1997–2005) and after MELD (2006–2012) in our state. A retrospective study was conducted including the data from all the liver transplant candidate waiting lists from July 1997 to December 2012. The data were related to the actual number of liver transplantations (Tr), the incidence of new patients on the list (I), and the number of patients who died while being on the waitlist (D) from 1997 to 2005 (the pre-MELD era) and from 2006 to 2012 (the post-MELD era). The number of transplantations from 1997 to 2005 and from 2006 to 2012 increased nonlinearly, with a clear trend to levelling to equilibrium at approximately 350 and 500 cases per year, respectively. The implementation of the MELD score resulted in a shorter waiting time until liver transplantation. Additionally, there was a significant effect on the waitlist dynamics in the first 4 years; however, the curves diverge from there, implying a null long-range effect on the waitlist by the MELD scores.