New multifactorial burn resuscitation formula offers superior predictive reliability in comparison to established algorithms.

UNLABELLED: The Parkland-Baxter formula is a widely utilized resuscitation guideline for the initial management of fluid deficits in burn victims. Implementation of resuscitation formulas has helped to reduce the incidence of shock and hypovolemic organ failure such as acute renal failure in the setting of burn trauma. However, it has been shown that indiscriminate implementation of these formulas may inappropriately suit individual patient's requirements. In our experience resuscitation by the Parkland formula often forced corrections in order to reach predefined resuscitation goals. OBJECTIVE: Given these findings we felt the need to refine formula based resuscitation strategies. PATIENTS AND MEASUREMENTS: Reviewing a subset of 81 burn admissions we screened for predictive parameters in addition to total body surface area burned (TBSA burned) and body weight influencing resuscitation volume requirements. DESIGN: Using multivariate linear regression analysis (MRA) various parameters were integrated in a stepwise forward mathematical selection procedure resulting in a modified resuscitation formula. MAIN RESULTS: A new formula including body weight, TBSA burned, inhalation injury (IHI), high blood alcohol level (BAL) and a compensating factor for advanced age was set up. The new formula was compared to the original Parkland formula. Both were assessed for predictive reliability (PR(+/-20%)). Using this strategy we were able to improve PR(+/-20%) from 28.4% to 51.9%. CONCLUSIONS: Optimal fluid resuscitation of severe burn victims is a complex clinical challenge. Rigid-formula based resuscitation schemes often fail to match all subtleties of current clinical practice but need to provide a reliable starting point for fluid resuscitation. We demonstrate a new multifactorial formula resulting in a better guide to initial fluid resuscitation.

Identification of novel proteins associated with hepatocellular carcinomas using protein microarrays.

Characterization of the protein profiles expressed by hepatocellular carcinomas (HCCs) may identify the genes involved in hepatocellular carcinogenesis and offers the possibility of elucidating clinical biomarkers. In an effort to discover such proteins and pathways that are deregulated in hepatocellular carcinogenesis, cellular proteomes of matched normal liver cells and carcinoma were analysed by tissue microdissection and protein microarrays. Using protein microarrays made up of 83 different antibodies, it was possible to monitor alterations of the protein levels in HCC and non-neoplastic liver tissue. Further analysis of altered proteins was performed using western blot analysis and tissue microarrays (TMAs) containing 210 HCC specimens and corresponding liver tissue. The protein microarray approach revealed differential expression between HCC and normal liver of 32 of the 83 proteins examined: 21 of these were up-regulated and 11 down-regulated. IGF (insulin growth factor) II, ADAM (a disintegrin and metalloproteases) 9, STAT (signal transducers and activators of transcription) 3, SOCS (suppressors of cytokine signalling) 3, and cyclin D1 were significantly up-regulated and collagen I, SMAD 4, FHIT (fragile histidine triad), and SOCS1 were down-regulated. The differential expression of these proteins was confirmed using western blot analysis and TMAs. Correlation of differentially regulated proteins with clinico-pathological data showed that cyclin D1 and SOCS1 were associated with tumour prognosis in univariate analysis, but not multivariate analysis. These data indicate that the development of an array-based approach for the determination of protein profiles in HCC may facilitate the identification of new proteins associated with carcinogenesis or prognosis.

Genetic and epigenetic alterations of the INK4a-ARF pathway in cholangiocarcinoma.

The INK4a-ARF locus, located on chromosome 9p21, encodes two cell-cycle regulatory proteins, p16(INK4a) and p14(ARF), acting through the Rb-CDK4 and p53 pathways. To study the contribution of each pathway in the tumourigenesis of cholangiocarcinoma, the alterations of p14(ARF), p16(INK4a), p53, and pRb were analysed. After microdissection, DNAs from 51 cholangiocarcinomas were analysed by methylation-specific PCR (MSP), restriction-enzyme related polymerase chain reaction (RE-PCR), microsatellite analysis, mRNA expression, and DNA sequencing. Immunohistochemistry of p14(ARF), p16(INK4a), p53, and pRb was also performed. Promoter methylation of p14(ARF) was found in 13/51 cases (25%) and p16(INK4a) showed aberrant promoter methylation in 39/51 cases (76%) which correlated with loss of mRNA transcription. Two tumours (4%) had homozygous deletion of the INK4a-ARF locus. Specific mutations of both exons were not detected. p14(ARF) inactivation appeared in the context of an unmethylated p16(INK4a) promoter in eight of 13 cases (61%) of the carcinomas methylated at p14(ARF). Mutations of p53 were found in 19 of 51 tumours (37%), and four of them (21%) harboured p14(ARF) inactivation. The pRb protein was detected in 30/51 (59%) tumours examined. The absence of pRB protein did not correlate with any of the examined parameters. Alterations of the INK4a-ARF locus, pRB or p53 status could not be established as independent prognostic factors in these tumours. These findings indicate that the INK4a-ARF locus is frequently inactivated in cholangiocarcinoma of the liver and occurs independently of the status of p53 or pRb.