Improving patient survival with the colorectal cancer multi-disciplinary team.

OBJECTIVE: There is little information on the impact of the colorectal multi-disciplinary team (MDT) in the United Kingdom. Our single operator presented his patients before and after the inception of an MDT meeting in June 2002. The aim of this study was to assess the effect of this on his patients' survival, and trends in the use of adjuvant chemotherapy. METHOD: Data were collected on all patients (n = 310) undergoing colectomy for colorectal cancer by one surgeon. Excluding patients with Dukes A stage, the pre-MDT cohort from January 1997 to May 2002 was 176 and the post-MDT cohort from June 2002 to December 2005 was 134. Three-year survival rates were calculated using Kaplan-Meier life table analysis. Prognostic factors were analysed using Cox-proportional hazard regression, and chemotherapy data analysed using the chi-squared test. Independent prognostic indicators of chemotherapy prescription were examined using binary logistic testing. RESULTS: MDT status was shown to be an independent predictor of survival on hazard regression analysis (P = 0.044). A significantly greater number of patients were prescribed adjuvant chemotherapy in the post-MDT cohort (P = 0.0002). MDT status was shown to be a significant prognostic indicator of chemotherapy prescription (P < 0.0001). Three-year survival for Dukes C patients was 58% in the pre-MDT group, and 66% in the post-MDT group (P = 0.023). CONCLUSION: There was a significant increase in patients undergoing adjuvant postoperative chemotherapy after the inception of the MDT. This was associated with a significant survival benefit in patients with Dukes C disease. The data suggest that the MDT process has resulted in an increase in the prescription of adjuvant chemotherapy, with 3-year survival being greater after its inception.

Patient maintained sedation for colonoscopy using a target controlled infusion of propofol.

In this study, we evaluated safety and recovery using a patient maintained, target controlled infusion of propofol for sedation in 20 patients undergoing colonoscopy. Using a handset with a two-minute lockout interval, patients could make 0.2 micro g.ml(-1) increments to an initial target plasma concentration of 1 micro g.ml(-1) up to a maximum 4.5 micro g.ml(-1). Four patients became oversedated but required no airway or circulatory interventions. Subjects had a significant reduction in mean (SD) heart rate: 78.7 (15) vs. 69.8 (13.5) (p < 0.001) and in systolic blood pressure 121.1 (13.2) mmHg vs. 96.5 (8.6) mmHg (p < 0.001). Choice reaction time testing 15 min after colonoscopy showed a significant median (IQR [range]) rise of 162 (- 16, 383.3 [-199-859]) ms (p < 0.05). Six patients had faster reaction times postcolonoscopy. All patients denied unpleasant recall and were satisfied with the system. Although oversedation was a problem in this model, we conclude that patient maintained propofol sedation could be possible for colonoscopy.

Selective targeting of a redox-active ubiquinone to mitochondria within cells: antioxidant and antiapoptotic properties.

With the recognition of the central role of mitochondria in apoptosis, there is a need to develop specific tools to manipulate mitochondrial function within cells. Here we report on the development of a novel antioxidant that selectively blocks mitochondrial oxidative damage, enabling the roles of mitochondrial oxidative stress in different types of cell death to be inferred. This antioxidant, named mitoQ, is a ubiquinone derivative targeted to mitochondria by covalent attachment to a lipophilic triphenylphosphonium cation through an aliphatic carbon chain. Due to the large mitochondrial membrane potential, the cation was accumulated within mitochondria inside cells, where the ubiquinone moiety inserted into the lipid bilayer and was reduced by the respiratory chain. The ubiquinol derivative thus formed was an effective antioxidant that prevented lipid peroxidation and protected mitochondria from oxidative damage. After detoxifying a reactive oxygen species, the ubiquinol moiety was regenerated by the respiratory chain enabling its antioxidant activity to be recycled. In cell culture studies, the mitochondrially localized antioxidant protected mammalian cells from hydrogen peroxide-induced apoptosis but not from apoptosis induced by staurosporine or tumor necrosis factor-alpha. This was compared with untargeted ubiquinone analogs, which were ineffective in preventing apoptosis. These results suggest that mitochondrial oxidative stress may be a critical step in apoptosis induced by hydrogen peroxide but not for apoptosis induced by staurosporine or tumor necrosis factor-alpha. We have shown that selectively manipulating mitochondrial antioxidant status with targeted and recyclable antioxidants is a feasible approach to investigate the role of mitochondrial oxidative damage in apoptotic cell death. This approach will have further applications in investigating mitochondrial dysfunction in a range of experimental models.

Bioenergetic consequences of accumulating the common 4977-bp mitochondrial DNA deletion.

Mutations and deletions in mitochondrial DNA (mtDNA) lead to a number of human diseases characterized by neuromuscular degeneration. Accumulation of truncated mtDNA molecules (delta-mtDNA) lacking a specific 4977-bp fragment, the common deletion, leads to three related mtDNA diseases: Pearson's syndrome; Kearns-Sayre syndrome; and chronic progressive external ophthalmoplegia (CPEO). In addition, the proportion of delta-mtDNA present increases with age in a range of tissues. Consequently, there is considerable interest in the effects of the accumulation of delta-mtDNA on cell function. The 4977-bp deletion affects genes encoding 7 polypeptide components of the mitochondrial respiratory chain, and 5 of the 22 tRNAs necessary for mitochondrial protein synthesis. To determine how the accumulation of delta-mtDNA affects oxidative phosphorylation we constructed a series of cybrids by fusing a human osteosarcoma cell line depleted of mtDNA (rho0) with enucleated skin fibroblasts from a CPEO patient. The ensuing cybrids contained 0-86% delta-mtDNA and all had volumes, protein contents, plasma-membrane potentials and mitochondrial contents similar to those of the parental cell line. The bioenergetic consequences of accumulating delta-mtDNA were assessed by measuring the mitochondrial membrane potential, rate of ATP synthesis and ATP/ADP ratio. In cybrids containing less than 50-55% delta-mtDNA, these bioenergetic functions were equivalent to those of cybrids with intact mtDNA. However, once the proportion of delta-mtDNA exceeded this threshold, the mitochondrial membrane potential, rate of ATP synthesis, and cellular ATP/ADP ratio decreased. These bioenergetic deficits will contribute to the cellular pathology associated with the accumulation of delta-mtDNA in the target tissues of patients with mtDNA diseases.

Alterations to glutathione and nicotinamide nucleotides during the mitochondrial permeability transition induced by peroxynitrite.

Peroxynitrite is a biologically important oxidant that damages mitochondria in a number of ways. We investigated the interaction of peroxynitrite with the mitochondrial glutathione pool by measuring the formation of oxidised glutathione and glutathione-protein mixed disulfides in mitochondria exposed to either peroxynitrite or tert-butylhydroperoxide. In contrast to tert-butylhydroperoxide, peroxynitrite converts 40-50% of mitochondrial glutathione to products other than disulfides, primarily higher oxidation states of sulfur. These data show that peroxynitrite interacts with mitochondria quite differently from oxidants commonly used in studying mitochondrial oxidative stress. Peroxynitrite also induces a permeability transition in the mitochondrial inner membrane, and here we show that this permeability transition is prevented by the NAD(P)H-linked substrates glutamate and malate and by the thiol reagent dithiothreitol. Glutamate and malate prevented complete oxidation of the NAD(P)H pool by peroxynitrite or tert-butylhydroperoxide but did not prevent oxidation of the mitochondrial glutathione pool or the formation of glutathione-protein mixed disulfides. This study is consistent with regulation of the permeability transition by critical protein thiol groups, whose redox state responds to that of the mitochondrial NAD(P)H pool, but which do not equilibrate directly with the mitochondrial glutathione pool.

Laparoscopic-assisted reversal of Hartmann's procedure: a simplified technique and audit of twelve cases.

Laparoscopic-assisted reversal of Hartmann's procedure was performed in 12 patients over a 2-year period. The group comprised three patients with colonic carcinoma and nine with diverticular disease, their mean age being 62 years (range 40-73). In all twelve cases, intestinal continuity was successfully restored, without conversion to open surgery, in a median anesthetic time of 165 min (range 110-240). One patient required a temporary defunctioning colostomy and two other patients had three complications. The median postoperative hospital stay was 8 days (range 5-12). We report a retrospective audit of this consecutive series of 12 patients, showing that laparoscopic-assisted reversal of Hartmann's is technically feasible. We also give details of the morbidity associated with the technique.

Laparoscopic conversion of polya gastrectomy to Roux-en-Y.

We describe a patient who underwent laparoscopic conversion of a Polya gastrectomy to a Roux-en-Y. We used the "Endo Stitch" (Autosuture Co., Ascot, UK) to simplify the creation of a laparoscopic anastomosis. The patient required minimal analgesia and made a rapid and uncomplicated recovery.

Bioassay of transforming growth factor-beta activity in acidic protein extracts from primary breast cancer specimens.

Acidic protein extracts have been made from breast tumour specimens, collected at the time of primary surgery. The extracts were partially purified by gel filtration and then tested for transforming growth factor-beta activity in a reproducible cell 3H-thymidine incorporation assay. Purified TGF-beta causes a reproducible increase in NRK colony formation and inhibits incorporation of 3H-thymidine by mink lung cells. However, some of the breast cancer extracts were stimulatory in the mink lung lung assay implying that a mitogenic factor like epidermal growth factor (EGF) was co-purified. Fourteen out of thirty extracts were scored positive for TGF-beta in the NRK colony forming assay and these tumours presented at an earlier clinical stage and were predominantly well differentiated.

T cell requirements for the rejection of renal allografts bearing an isolated class I MHC disparity.

This study has examined the cellular and humoral responses underlying the rejection of rat renal allografts bearing an isolated RT1Aa class I MHC disparity. RT1Aa disparate kidneys were rejected promptly by high responder RT1u but not by low responder RT1c recipients (median survival time 10 d and greater than 100 d, respectively). The magnitude and phenotype of the cellular infiltrate were similar in rejecting and nonrejecting RT1Aa disparate kidneys. Paradoxically, graft infiltrating cells and spleen cells from RT1u recipients showed minimal ability to lyse donor strain lymphoblasts in vitro, whereas effector cells from RT1c recipients showed modest levels of cytotoxicity. Injection of RT1u rats with MRC OX8 mAb was highly effective at selectively depleting CD8+ cells from graft recipients but had no effect in prolonging the survival of RT1Aa disparate grafts despite the complete absence of CD8+ cells from the graft infiltrate, which included numerous CD4+ T cells and macrophages. RT1u, but not RT1c, recipients mounted a strong alloantibody response against RT1Aa disparate kidneys. Immune serum obtained from RT1u recipients that had rejected a RT1Aa disparate graft was able, when injected into cyclosporin-treated RT1u recipients, to restore their ability to reject a RT1Aa, but not a third-party RT1c, kidney. These results suggest that CD8+ cells in general and CD8+ cytotoxic effector cells in particular are unnecessary for the rapid rejection of RT1Aa class I disparate kidney grafts by high responder RT1u recipients. By implication, CD4+ T cells alone are sufficient to cause prompt rejection of such grafts and they may do so by providing T cell help for the generation of alloantibody.

Gas chromatography-mass spectrometry and the measurement of vitamin D metabolites in human serum or plasma.

Although methods for the measurement of vitamin D metabolites continue to be developed, few have been properly validated by comparison with methods based on gas chromatography-mass spectrometry, widely accepted as being the definitive methodology. To the best of our knowledge, only three such comparisons have been carried out (14, 42, 83), all three examining HPLC assays for 25-OH-D. This lack of proper validation leads to lack of certainty as to the specificity of many assays widely used for clinical investigation. In our view there is an obvious need for the continuing development of mass fragmentographic assays for vitamin D and its metabolites, primarily for use as reference procedures for the evaluation of less rigorous methodologies. Provided standards, both labeled and unlabeled, become more widely available, development of specific mass fragmentographic assays for any metabolite of vitamin D should be possible. For metabolites where no specific binding protein or antiserum is available, mass fragmentography may be the only alternative.

Herpes simplex virus reactivation in surgical patients.

Evidence for oral shedding of herpes simplex virus Type I (HSV) indicating endogenous viral reactivation as a measure of depressed host defense was sought in 44 critically ill surgical patients. Eighteen (41%) of these showed persistent HSV shedding. None of 50 controls showed HSV shedding, but 4 (10%) of 42 patients undergoing elective surgery showed transient postoperative viral reactivation. In the critically ill surgical patients, oral HSV shedding was not related to outcome. However, failure to develop a rise in specific HSV antibody, in the presence of viral shedding, was associated with a high mortality. This is further evidence for impairment of both cell-mediated and humoral immunity in critically ill surgical patients.

Improved vascular access for patients receiving chemotherapy.

Indwelling central venous catheters were used for vascular access in 25 oncology patients. The lines were used for sampling, administration of blood products, chemotherapeutic agents, parenteral nutrients and occasionally plasmapheresis. The complication rate was no higher than in reported series in which the catheters were reserved for parenteral nutrition. We believe that a central venous cannula can be safely used as the sole means of vascular access in those patients with consequent psychological and practical benefits.