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Backgrounds. Limited effective palliative treatments exist for pancreatic cancer which includes surgery or chemotherapy. Radiofrequency ablation (RFA) uses high frequency alternating current to ablate diseased tissue and has been used to treat various tumors. In this study, we evaluated a prototype probe adjusted to the EUS-needle to perform EUS-RFA to permit coagulative necrosis in the pancreas.Methods. Five Yucatan pigs underwent EUS-guided radiofrequency ablation of the head of their pancreas. Using an EUS-needle, RFA was applied with 6 mm and then 10 mm of the probe exposed at specific wattage for preset durations.Results. Only one pig showed moderate levels of pancreatitis (20% proximal pancreatitis). The other animals showed much lower areas of tissue damage. In 3 of the 5 pigs, the proximal pancreas showed greater levels of tissue injury than the distal pancreas, consistent with the proximity of the tissue to the procedure site. In 1 pig, both proximal and distal pancreas showed minimal pancreatitis (1%). There was minimal evidence of fat necrosis in intra-pancreatic and/or extra-pancreatic adipose tissue.Conclusion. EUS-guided RFA of the pancreatic head with the monopolar probe through a 19-gauge needle was well tolerated in 5 Yucatan pigs and with minimal amount of pancreatitis.

Background and Objectives: Hematopoietic stem cells (HSC) are the most widely studied and characterized adult stem cells, which play an essential role in sustaining the formation of blood and immune system. The ease of their manipulation, the lack of serious ethical issues, and, in the autologous setting, the absence of their immunogenicity, have made them an attractive tool for developing stem cell-based therapies. Exploiting the properties of HSC by microRNA (miRNA) profiling offers an attractive approach to identify new regulators of stem cell fate. Although numerous miRNA have been screened from hematopoietic stem cells (HSC), the targets corresponding to many of these miRNA have not yet been fully elucidated. Therefore the objective of this study is to generate a miRNA profile from a subpopulation of adherent CD34+ HSC isolated from G- colony-stimulating factor mobilized peripheral blood aiming to understand the role of selected miRNA in regulating HSC stemness. Methods: CD34+ cells from patients' blood were isolated using a CD34+ isolation kit (Miltenyi Biotec) according to the manufacturer's protocol. miRNA profiling of adherent and nonadherent CD34+ cells was done using TaqMan Array MicroRNA Cards. Nanog expression levels was tested using a dual-luciferase reporter construct for miR-181a* or its mutant variant and Nanog 3′ UTR mRNA. Results: In this study, we have identified eight clusters of miRNA that were differentially expressed in an adherent subpopulation of CD34+ stem cells. Further analysis of one of the clusters by bioinformatics revealed that a miRNA, miR-181a*, which is highly expressed in the adherent CD34+ cells, affects the expression levels of Nanog, a stem cell surrogate marker. We show specifically by reporter assay and mutational analysis that miR-181a* targets a seedless 3′ compensatory site in the 3′UTR of Nanog and affects gene expression. We demonstrate that inhibiting miR-181a* upregulates the Nanog expression level, in addition to an increase in alkaline phosphatase activity. Conclusions: In conclusion, our results highlight a new stem cell-related target for the miR-181 family and show that miR-181a* directly targets Nanog in a subpopulation of CD34+ stem cells suggesting a possible role of miR-181a* in regulation of adherent CD34+ HSC cells stemness.

Maintaining a balanced prostate microenvironment is pivotal for normal development and homeostasis of the prostate gland. This balance however is severely disrupted during the progression of prostate cancer where the local microenvironment becomes compromised. The cellular components associated with the microenvironment, including stromal cells, immune cells, blood vessels, and the extracellular matrix, interact cooperatively with prostate cancer cells through paracrine and autocrine actions of soluble growth factors and cytokines thus creating a modified tumour microenvironment. Understanding how paracrine and autocrine factors interact in this microenvironment may lead to improved understanding of prostate cancer progression and to the development of drug combinations that might target both the primary and metastatic prostate cancer tumour microenvironments.

It is now recognized that small noncoding RNA sequences have the ability to mediate transcriptional activation of specific target genes in human cells. Using bioinformatics analysis and functional screening, we screened short-activating RNA (saRNA) oligonucleotides designed to target the promoter regions of the pluripotency reprogramming factors, Kruppel-like factor 4 (KLF4) and c-MYC. We identified KLF4 and c-MYC promoter-targeted saRNA sequences that consistently induced increases in their respective levels of nascent mRNA and protein expression in a time- and dose-dependent manner, as compared with scrambled sequence control oligonucleotides. The functional consequences of saRNA-induced activation of each targeted reprogramming factor were then characterized by comprehensively profiling changes in gene expression by microarray analysis, which revealed significant increases in mRNA levels of their respective downstream pathway genes. Notably, the microarray profile after saRNA-mediated induction of endogenous KLF4 and c-MYC showed similar gene expression patterns for stem cell- and cell cycle-related genes as compared with lentiviral vector-mediated overexpression of exogenous KLF4 and c-MYC transgenes, while divergent gene expression patterns common to viral vector-mediated transgene delivery were also noted. The use of promoter-targeted saRNAs for the activation of pluripotency reprogramming factors could have broad implications for stem cell research. Copyright © 2012 American Society of Gene & Cell Therapy. Published by Elsevier Inc. Attribution-NonCommercial-NoDerivs 3.0 Unported (CC BY-NC-ND 3.0)

U functional loss of insulin producing islet β-cells, some diabetic patients become dependent on life-long insulin supplementation therapy. Bioengineering surrogate insulin producing cells is an alternative replacement strategy. We have developed a novel cell culture methodology using RNA oligonucleotides to differentiate adult human hematopoietic CD34+ cells into insulin secreting cells. By transfecting RNA to increase transcript levels of the master regulator of insulin biosynthesis, v-maf musculoaponeurotic fibrosarcoma oncogene homolog A (MafA), several pancreatic endodermal genes were upregulated during the differentiation procedure. These included Pancreatic and duodenal homeobox gene-1 (PDX1), Neurogenin 3, NeuroD, and NK6 homeobox 1 (NKx6.1). Differentiated CD34+ cells also expressed glucokinase, Glucagon-like peptide-1 (GLP1), Sulfonylurea receptor-1 (SUR1) and phogrin all essential for glucose sensitivity and insulin secretion. The differentiated cells appropriately processed c-peptide and insulin in response to increasing glucose stimulation as shown by enzyme-linked immunosorbent assay (ELISA), fluorescence activated cell-sorting (FACS) analysis, Western blotting and immunofluorescence staining. We provide a new approach to exploit human CD34+ cells that may be useful in developing insulin producing surrogate cells for treating diabetes.

Laparoscopic surgery has undergone a major transformation in the last two decades, with the advancement of new instruments, new techniques, and increased surgical experience and skill. Laparoscopic hepatic surgery is now not only feasible and safe, but depending on the indication, preferable, decreasing perioperative morbidity and hospital stay. Nevertheless, laparoscopic liver resection remains a highly challenging surgical procedure because of the risk of massive bleeding during liver transection and the complicated biliary and vascular anatomy of the liver. The Laparoscopic Habib™ 4X is a device specifically designed to address these issues. The device and technique of hepatic transection using the Laparoscopic Habib™ 4X are described.

Video abstract Video, Stroke is a leading cause of death and disability worldwide. Stem cell therapy is an emerging therapeutic modality with evidence of significant benefits in preclinical stroke models. A number of phase I and II clinical trials have now been completed, with several more currently under way. Translation to the bedside, however, remains a long way off, and there are many questions that remain unanswered. This review will summarize the current evidence and ongoing clinical trials worldwide, and explore the challenges to making this a realistic treatment option for the future.

Molecular genetic changes are better characterized in colorectal carcinoma than in other common adult tumours. Consistent allele losses, or loss of heterozygosity (LOH), on chromosomes 5q, 17p and 18q have been well established. These changes are associated with the prognosis of the disease. Little is known of such changes in liver metastases of colorectal origin. The extent of allelic loss and its association with clinical features were investigated in 19 patients with colorectal liver metastases by using 24 probes to detect restriction fragment length polymorphism. A high frequency of LOH on chromosomes 5q, 17p and 18q was found in these secondary tumours. No consistent loss has so far been shown in any other chromosome. The frequency of allele loss correlated significantly with prognostic features such as the number and size of liver secondaries (P < 0·005), metastasis to the lymph nodes (P < 0·01) and curative or palliative operation (P < 0·02).

A segment of 712 bases coding for part of the human stearoyl-CoA desaturase gene was made by polymerase chain reaction (PCR) using primers based on published rat cDNA sequences. The human PCR product was confirmed by DNA sequencing. It was next cloned into a vector from which anti-sense, highly radioactive RNA transcripts were made in vitro using T7 polymerase. The transcripts were used to probe desaturase mRNA in a number of human tumour and control tissues, using a very sensitive solution hybridization/RNase protection assay. Increased desaturase mRNA levels were found in colonic and oesophageal carcinomas and in hepatocellular adenoma; however, no consistent trend was seen in hepatocellular carcinoma. It is suggested that certain classes of tumour may exhibit increased levels of desaturase mRNA.

Hepatoblastoma is a rare cancer, but it is the most common primary malignant hepatic tumour in childhood, accounting for 71-75°/,, of liver cancers (1,2). The environmental risk factors for this embryonal tumour are not known. The risk is unrelated to maternal oestrogen exposure, alcohol consumption and smoking (1). For hepatocellular carcinoma (HCC), the most common primary liver malignancy in adults, hepatitis B virus (HBV) infection is a major risk factor (3), while hepatoblastoma is not associated with HBV infection (1,4). Recently, much progress has been made in detailing the genetic-molecular changes of the tumour.

Background Surgical resection remains the gold standard in dealing with liver tumours. Blood loss, biliary leak and postoperative liver function are still the main concerns of surgeons operating on the liver, even though different techniques have been developed to allow safer liver resection. A novel concept for liver resection is described using a radiofrequency energy (RF) assisted technique. Method A patient with a large colorectal liver metastasis located in segments VI, VII, VIII underwent a right hepatectomy using this technique. At laparotomy the tumour was staged with intraoperative ultrasonography, and a ‘cooled tipped’ radiofrequency probe was used to achieve a ‘zone of desiccation’ in the liver parenchyma 2 cm away from the edge of the tumour. Liver parenchyma was subsequently divided with a surgical scalpel. Results The resection time was 80 min with a blood loss of 30 ml. The patient was discharged on the ninth postoperative day without complications. Discussion Liver resection assisted by RF energy is feasible and safe. This technique could offer a new method for ‘transfusion-free’ resection without the need for sutures, ties, staples, tissue glue or admission to the intensive care unit.

The effects of ablation with various settings of powers and times using a newly developed radiofrequency (RF) ablation device, the Habib™ EndoHPB catheter, are not well known. In the present study, we examined the effects of a novel RF ablation catheter using resected fresh pig livers and evaluated the macro- and microscopic effects of RF ablation under various conditions.The RF application was performed step by step at 5, 10, 15, and 20 W power and 60, 90, 120 s, respectively. Macroscopic and microscopic findings of the ablation area were evaluated at each setting.The mean lengths of the short axis of the ablation area at 10 W and 60, 90 and 120 s were 8.0 ± 1.0, 8.3 ± 1.2, and 9.7 ± 0.6 mm, respectively. The mean lengths of the long axis at 10 W power and 60, 90 and 120 s were 20.3 ± 0.6, 21.3 ± 1.6, and 28.3 ± 2.1 mm, respectively. Although the lengths of the short and long axes at 5 and 10 W increased gradually with power, there were no obvious differences in either short or long axis lengths between 15 and 20 W. Of all the settings, only at 5 W and 60 and 90 s did the long axis of the ablation show separate areas around the 2 ring electrodes.Although other sequelae including hemorrhage, pancreatitis, acute inflammatory changes, perforation and late fibrosis could not be investigated in our ex-vivo pig model, our study clarified the relationship between ablation powers and times and the effects concerning depth and longitudinal spread of ablation. Although the clinical ablation setting at 7-10 W power and 2 min is suitable, ultimately the ablation power and time should be adjusted according to the size of masses using examples from the present results.

To develop a new rabbit model of arterial stenosis using endovascular radiofrequency (RF) energy.Ten rabbits were used for multiple endovascular RF applications to the aorta and left common carotid artery through the Habib™ VesCoag™ catheter. Angiography and color Doppler ultrasound were used to assess vessel patency immediately following the procedure and six weeks later. One rabbit was sacrificed following the procedure for histopathologic analysis of the vessel wall. Two rabbits died of aortic and carotid rupture, respectively, immediately after the procedure. The remaining seven rabbits were sacrificed after six-week follow-up for histopathological analysis.Optimal RF generator settings to induce significant arterial stenosis (50%) without complications were standardized at 24-26 watts (W) for 1.5 min for the aorta and 6 W for 1 min for the common carotid artery. The six-week follow-up showed permanent results in all surviving rabbits. Histopathology revealed intima and medial smooth muscle layer necrosis.We have developed a novel rabbit model of arterial stenosis using endovascular RF energy. Our model is fast, safe, inexpensive, and reproducible. It would be useful for experimental investigations and new therapeutic devices.

Hepatocellular carcinoma (HCC) is the fifth most common neoplasm in the world, closely correlated with viral hepatitis and liver cirrhosis. The vast majority of HCC patients present at a late stage and are unsuitable for surgery due to limited liver functional reserve. Tumors can involve major vessels or hilar structures, necessitating major liver resection and/or rendering liver resection unfeasible. A series of new technologies have been developed to optimise HCC management. Stem cell therapy improves impaired liver functional reserve prior to liver resection. Intravascular radiofrequency ablation recanalises the portal vein invaded by tumour thrombus and endobiliary radiofrequency ablation restores and extends biliary patency of the bile duct invaded by malignancy. Laparoscopic radiofrequency assisted liver resection minimizes blood loss and avoids liver warm ischemia, while increasing parenchymal sparing. These benefits combined maximize the safety of liver resection.

The feasibility and outcome of large volume injection of gene solution in a segment of a liver lobe, without backflow, were studied in a porcine model, using a custom-designed balloon catheter.Eight anesthetized pigs underwent successful injection of 200 ml of gene solution at a rate of 20 ml/s via a minimally invasive technique without backflow. A custom-made balloon catheter was introduced under fluoroscopy guidance into the right lateral liver lobe via the right external jugular vein. The vein of the liver lobe was occluded with the balloon catheter and contrast material was injected to check if total occlusion was achieved. Since there was no backflow an angiographic pump injected the solution. The catheter was left in place for 10 min. Then contrast material was injected to check whether the vein remained occluded.All animals tolerated the procedure without obvious adverse effects. Ultrasound scan showed no gross changes within liver three days following the infusion. A transient rise in platelet count was observed which returned to normal after 13 days and remained stable; all other biochemistry values were normal.Injecting large volume of gene solution in a liver lobe segment using this minimally invasive technique in a porcine model is possible, making the development of a successful gene transfer protocol in humans feasible.

It has been established that loss of tumour suppressor genes is crucial in carcinogenesis. There has been no reported study on searching for tumour suppressor genes in cholangiocarcinomas as yet. In order to investigate the loss of heterozygosity (LOH), which may represent such gene loss, in cholangiocarcinoma, we studied 14 patients with this tumour using restriction fragment length polymorphism analysis. Twenty-two probes assigned to chromosomes 1, 5, 7, 9, 11, 12, 13, 14, 16, 17 and 18 were used. Allelic losses were found in chromosomal regions 5q35-qter and 17p13. Loss of genetic material in these regions in cholangiocarcinoma was shared with hepatocellular carcinoma. Probes for other chromosomes have as yet shown no consistent LOH. In conclusion, this study for the first time showed LOH on chromosomes 5 and 17 in cholangiocarcinoma. Images Figure 1

1. Isolated perfused cirrhotic rat livers were used to study the effects of an increase in portal perfusion pressure and portal flow on the microcirculation and viability of the hepatocytes. Cirrhosis was induced by CCl4, and Krebs-Ringer bicarbonate buffer solution was used as the perfusate. Portal perfusion pressures were increased incrementally between 25 and 45 cm H2O. The viability of the livers was assessed and histological studies were performed under light and electron microscopy. 2. An increase in portal perfusion pressure induced an increase in hepatic flow in all the experiments (P < 0.05). Hepatic flow was 2.52 ml min−1 g−1 of liver (SD 0.67; n = 5) at basal pressure compared with 4.19 ml min−1g−1 of liver (SD 0.93; n = 5) and 5.91 ml min−1g−1 of liver (SD 0.63; n = 5) when pressures were raised to 25 and 45 cm HzO, respectively. Portal perfusion pressure and hepatic flow were correlated (r = 0.908; P < 0.001; n = 30). 3. Production of the enzyme alanine aminotransferase (EC 2.6.1.2) increased significantly from 5.69 i.u. ml−1min g-l of liver (SD 3.62; n = 5) to 23.53 i.u. ml−1min g−1 of liver (SD 16.7; n = 5) when the perfusion pressure was raised from baseline to 30 cm H2O. In all the cases the porto-caval gradient of enzyme production was within the normal range. No correlation existed between the release of enzyme and portal perfusion pressures 4. Histological studies demonstrated sinusoidal dilatation in one case out of five in the control group compared with nine cases out of 25 in the high perfusion pressure groups. Dead sinusoidal cells coloured by Trypan Blue were observed in one case out of five in the control group compared with nine cases out of 25 in the high perfusion pressure groups. With electron microscopy, occasional lesions such as detachment of endothelial cells or blebs were found. No relationship existed between portal perfusion pressures and histological lesions. 5. This study shows that a brief increase in portal perfusion pressure in the isolated perfused cirrhotic rat liver induces an increase in hepatic flow without a change in the viability of the liver and without significant histological alterations.

The only curative procedure to date for liver tumors is surgical resection, which remains a major procedure with marked morbidity and mortality. Radiofrequency (RF) has increasingly been used for both ablation and resection. On the basis of this technique, a new bipolar RF device, Habib 4X, has been developed and used clinically. We present our technique of liver resection with this device in a patient with colorectal liver metastases.A patient with situs inversus who had colorectal liver metastases in her left lobe underwent left lateral segmentectomy with the new device, a four-electrode bipolar resection device that uses RF energy for tissue necrosis. After laparotomy and intraoperative ultrasound, the plane resection was marked 1 cm away from the edge of the lesion. Coagulative desiccation was performed along this plane using this sealer connected to a RF generator. The necrosed band of parenchyma was then divided with a scalpel and resection completed.The length of the procedure was 105 minutes; resection time was 35 minutes. Total blood loss was 100 ml. No blood transfusions were required, and the patient was not admitted to the intensive care unit after surgery. The patient was discharged 10 days after surgery without any surgical complications.We think that RF-assisted liver resection with this new device is safe and effective. It is quicker than conventional RF and may reduce overall hospital stay in liver resection patients.

Two-stage liver resection (2-SLR) is used clinically in conjunction with portal vein embolization for bilobar disease to increase the number of patients suitable for liver resection. The long-term outcomes after 2-SLR for multiple bilobar colorectal liver metastases (CLM) was examined. Patients who sought care between November 2003 and April 2006 with multiple CLM considered suitable for 2-SLR were prospectively followed. Clinicopathological data were collected. Surgical outcomes were defined as complete clearance of tumor (R0/R1/R2), postoperative morbidity (within 3 months), 30 day mortality, disease-free survival (DFS), and overall survival (OS). A total of 131 patients with CLM underwent liver resection during the study period, 38 of whom were planned for a 2-SLR for multiple bilobar disease. Only 33 (87%) completed the 2-SLR with a curative intent. Five patients did not undergo stage II resection because of disease progression. The postoperative morbidity was 11 and 33% after stage I and stage II liver resections, respectively. Five patients (13%) encountered postoperative complications specific to liver surgery. The median interval from stage II resection to disease recurrence in the R0 group was 18 months versus 3 months in the R1/R2 group (P

The existence of cancer stem cells continues to be extensively debated among scientists, and given the mounting evidence, it is an area that needs further examination. The possible reason for resistance to current treatment modalities in cancer is that we have yet to discover a method to eradicate cancer stem cells. This chapter describes the theory of cancer stem cells, portrays the role of cancer stem cells in radiation and chemotherapy resistance, and describes methods for their isolation. Finally, it presents current knowledge on cancer stem cells in solid organ tumors and discusses the limitations of current isolation strategies.

Background The purpose of this study was to present a radiofrequency (RF) endovascular occlusion device (ie, Habib VesCoag Catheter; EMcision Ltd, London, UK) and to evaluate safety and efficacy of the device for complete occlusion of normal porcine vessels. Methods The study included 20 pigs. In each pig, a segmental branch of the right hepatic artery, a branch of the splenic artery, and a branch of one of the renal arteries were catheterized. A single or multiple applications of RF energy were performed until vessel occlusion was achieved. Fifteen days later, angiography was repeated to assess vessel patency. The vessels were then excised for pathological analysis. Results Vessels 2.5 to 6 mm in diameter were treated. Complete occlusion with a single attempt was achieved using a mean amount of energy of 110.67 J in vessels 2.5 to 3 mm, 111.67 J in vessels 3.5 to 4 mm, and 116.63 J in vessels 5 to 6 mm in diameter and was confirmed by angiography at the 15-day follow-up. Conclusions Vascular occlusion can be effectively and safely achieved by endovascular application of RF energy to normal porcine arteries using the Habib VesCoag catheter.

IF : 4,104; International audience; In vivo gene transfer to the human respiratory tract by adenovirus serotype 5 (Ad5) vectors has revealed their limitations related to inefficient gene transfer, host antiviral response, and innate adenoviral toxicity. In the present work, we compared the cytotoxicity and efficiency of Ad5 and a chimeric Ad5F35 vector with respect to CFTR gene transfer to cystic fibrosis (CF) and non-CF human airway epithelial cells. We found that high doses of Ad5 vector had an adverse effect on the function of exogenous and endogenous CFTR. Results obtained with Ad5 capsid mutants suggested that the RGD motifs on the penton base capsomers were responsible for the negative effect on CFTR function. This negative interference did not result from a lower level of biosynthesis and/or altered cellular trafficking of the CFTR protein, but rather from an indirect mechanism of functional blockage of CFTR, related to the RGD integrin-mediated endocytic pathway of Ad5. No negative interference with CFTR was observed for Ad5F35, an Ad5-based vector pseudotyped with fibers from Ad35, a serotype that uses another cell entry pathway. In vitro, Ad5F35 vector expressing the GFP-tagged CFTR (Ad5F35-GFP-CFTR) showed a 30-fold higher efficiency of transduction and chloride channel correction in CFTR-deficient cells, compared with Ad5GFP-CFTR. Ex vivo, Ad5F35-GFP-CFTR had the capacity to transduce efficiently reconstituted airway epithelia from patients with CF (CF-HAE) via the apical surface, restored chloride channel function at relatively low vector doses, and showed relatively stable expression of GFP-CFTR for several weeks.

Primary liver cancer is amongst the commonest tumors worldwide, particularly in parts of the developing world, and is increasing in incidence. Over the past three decades, surgical hepatic resection has evolved from a high risk, resource intensive procedure with limited application, to a safe and commonly performed operation with a range of indications. This article reviews the approach to surgical resection for malignancies such as hepatocellular cancer, metastatic liver deposits and neuroendocrine tumors. Survival data after resection is also reviewed, as well as indications for curative resection.

Little is known of the molecular-genetic changes in carcinoma of the pancreas (CaP). In order to investigate the allele loss, or loss of heterozygosity (LOH), in CaP, we studied 13 patients with exocrine CaP and two with endocrine CaP using restriction fragment length polymorphism analysis. Twenty probes assigned to chromosomes 1, 5, 7, 9, 11, 12, 13, 14, 16, 17 and 18 were used. The frequency of LOH, or fractional allele loss (FAL), was found in two endocrine tumours to be 0.333 and 0.455 respectively; and FAL in 13 oxocrine tumours ranged from 0 to 0.25. Allele loss was shown in both exocrine and endocrine tumours by the probes Lambda MS1 at 1p33-35, and pMS51 at 11q13. Probes for other chromosomes have as yet shown no consistent LOH. In conclusion, the study showed LOH on chromosomes 1 and 11 in both exocrine and endocrine CaP. Images Figure 1

The aim of this study was to investigate the hypothesis that saturated fatty acids are differentially cytotoxic to cancer cells. Three studies were undertaken to: (1) measure the toxicities of stearic and oleic acids to normal and malignant cells in vitro, (2) assess if there is any relationship between toxicity and relative fatty acid composition and (3) determine whether the relative fatty acid composition of a cancer cell line could be modified by sterculic acid, an inhibitor of delta-9-desaturase. Stearic (18:0) and oleic (18:1) acids inhibited the colony-forming abilities of five human cancer cell lines and two non-neoplastic cell lines in a dose-dependent fashion. The concentration of oleic acid required to reduce colony formation ability by 50% was 2.5-6.0-fold greater than that of stearic acid. Addition of sterculic acid to a cancer cell line resulted in steady-state levels of stearic acid and increasing percentage of oleic acid.

Purpose There are few data on the impact of immediate and differing surgical interventions on circulating tumor cells (CTCs), nor their compartmentalization or localization in different anatomic vascular sites. Patients and Methods CTCs from consecutive patients with colorectal liver metastases were quantified before and immediately after open surgery, laparoscopic resection, open radiofrequency ablation (RFA), or percutaneous RFA. For individuals undergoing open surgery, either hepatic resections or open RFA, CTCs were examined in both systemic and portal circulation by measuring CTCs in samples derived from the peripheral vein, an artery, the hepatic portal vein, and the hepatic vein. Results A total of 29 consecutive patients with colorectal liver metastases with a median age of 55 years (range, 30 to 88 years) were included. CTCs were localized to the hepatic portosystemic macrocirculation with significantly greater numbers than in the systemic vasculature. Surgical procedures led to a statistically significant fall in CTCs at multiple sites measured. Conversely, RFA, either open or percutaneous, was associated with a significant increase in CTCs. Conclusion Surgical resection of metastases, but not RFA, immediately decreases CTC levels. In patients with colorectal liver metastases, CTCs appear localized to the hepatic (and pulmonary) macrocirculations. This may explain why metastases in sites other than the liver and lungs are infrequently observed in cancer.

Segmental hepatectomy is appealing for several reasons including preservation of liver parenchyma, reduction of intraoperative blood loss, and blood replacement by dividing tissues along the anatomic planes. A simple technique guided by intraoperative ultrasound is described here using radio-frequency energy to create coagulative desiccation of segmental or subsegmental arterial and portal vessels. Thirty patients underwent a segmental resection using this technique without mortality and with minor morbidity. This technique has a major advantage of being easy and safe to apply. We believe it has a potentially important role in both open and laparoscopic liver surgery.

Lung cancer is the leading cause of death from cancer in the world. Although the molecular network of lung carcinogenesis has been partly known at the levels of genes and proteins, and personalized therapy based on the genetic changes has made considerable progress in the last decade, the high mortality rate is not markedly changed. MicroRNAs (miRNAs), a class of short endogenous RNAs, acting as post-transcriptional regulators of gene expression, are similar with siRNAs in both the biosynthesis and the function steps. While, miRNAs mostly silence gene expression by binding imperfectly matched sequences in the 3' UTR of target mRNA, which is different with siRNAs by targeting ORF of mRNA with a perfectly complementary manner. miRNAs have multiple functions in lung development, and abnormal expression of miRNAs could lead to lung tumorigenesis. The different expression profiles of miRNAs in lung cancer, and the stability of miRNAs in serum, all together make them as new potentially clinical biomarkers for diagnosis and prognosis. Moreover, miRNAs may serve as either novel potential targets acting directly as oncogenes (e.g. miR-17-92 cluster) or directly therapeutic molecules working as tumor suppressor genes (e.g. let-7 family). RNAi technology based on miRNAs has many advantages over siRNAs, such as in vivo stability, highly RNA promoter-compatibility and no overt toxicity. Eventually, it might overcome the present disadvantages and become a good candidate for lung cancer therapy.

A 46-yr-old female with hepatocellular carcinoma and severe hepatitis B-related liver cirrhosis received a domino liver graft from a 25-yr-old female with homozygous familial hypercholesterolemia (HFHC) in September 2001. Hypercholesterolemia occurred in the graft recipient within one yr after transplantation and was partially controlled by atorvastatin. Three yr after transplantation, an autologous CD34+ cell transplantation was performed in order to better control the hypercholesterolemia. Only preliminary results of this domino liver transplantation (DLT) were published in 2003, without a long-term analysis of the hypercholesterolemic effects in recipient. Subsequent to DLT, the average plasma cholesterol level in the domino donor rapidly normalized and seven yr after had a value of 182 mg/dL. After seven-yr follow-up, the domino recipient has no hepatocarcinoma recurrence. Moreover, no signs of cardiovascular or atherosclerotic lesions were noted despite an elevated plasma cholesterol level (339 mg/dL after seven yr of follow-up) resistant to drug therapy and stem cell autotransplantation. In conclusion, DLT using a liver graft from a patient with HFHC provides a viable option for marginal recipients.

Suppressor gene loci involved in the development of hepatocellular carcinoma (HCC) have not been fully identified. The aim of this study was to look for consistent allele loss, or loss of heterozygosity (LOH), in HCC which might represent such gene loci. We have prepared DNA from tumour and non-tumour material from 16 patients with HCC (nine with and seven without liver cirrhosis). Tumour DNA was compared with non-tumour DNA by Southern analysis performed with a panel of 22 probes recognising restriction fragment length polymorphisms assigned to chromosomes 1, 4, 5, 7, 9, 11, 12, 13, 14, 16, 17, 18 and 20. Non-tumour DNA from five of the seven patients with HCC without cirrhosis was heterozygous with the probe Lambda MS8 (5q35-qter), and in all five there was LOH in tumour DNA. Probes for other regions of chromosome 5 have as yet shown no LOH in this group of patients. Cirrhotic HCC patients exhibited LOH on chromosomes 1q and 5p but not in the region 5q35-qter. Both groups of HCC showed LOH on chromosome 17p13. Screening with other probes has not shown any consistent LOH in either group as yet. A comparison of LOH on chromosome 5 in seven patients with colorectal metastasis in the liver showed a different pattern, which suggests that the proposed tumour suppressor gene locus for HCC without cirrhosis on chromosome 5 appears to be distinct from the familial adenomatous polyposis coli gene. Images Figure 1

Background Radio-frequency-assisted liver resection has been shown to allow virtually bloodless procedures without the need for vascular exclusion manoeuvres. Our primary end-point was to evaluate safety and feasibility of RF-assisted liver resection in cirrhotic patients with hepatocellular carcinoma. Our second end-point was to assess whether the RF-assisted procedure influence the outcome in terms of morbidity and mortality. Methods A retrospective study was done of 55 cirrhotic patients who underwent RF-assisted liver resection for HCC at our Departments between September 2001 and October 2007. Results In the period of study 29 monosegmentectomies, 20 bisegmentectomies, 4 trisegmentectomies and 2 right hepatectomies were carried out. Vascular exclusion manoeuvres were never performed. One patient died post-operatively because of untreatable hepatorenal syndrome. Twelve patients out of 55 experienced 21 complications including complicated pleural effusion, intra-abdominal collection, hepatorenal syndrome, ascites, hematoma and biliary fistula. Post-operative liver failure occurred in one case (1.8%). In patients without post-operative morbidity (n = 43) the 4-year survival rate was 62% versus 24% in patients who experienced post-operative morbidity (n = 12) (P = 0.02). Conclusions RF-assisted liver resection is a safe and feasible procedure associated with lower morbidity and hospital mortality rates even in case of liver cirrhosis. J. Surg. Oncol. © 2008 Wiley-Liss, Inc.

Intranuclear crystalline inclusions have been observed in the nucleus of epithelial cells infected with Adenovirus serotype 5 (Ad5) at late steps of the virus life cycle. Using immuno-electron microscopy and confocal microscopy of cells infected with various Ad5 recombinants modified in their penton base or fiber domains, we found that these inclusions represented crystals of penton capsomers, the heteromeric capsid protein formed of penton base and fiber subunits. The occurrence of protein crystals within the nucleus of infected cells required the integrity of the fiber knob and part of the shaft domain. In the knob domain, the region overlapping residues 489–492 in the FG loop was found to be essential for crystal formation. In the shaft, a large deletion of repeats 4 to 16 had no detrimental effect on crystal inclusions, whereas deletion of repeats 8 to 21 abolished crystal formation without altering the level of fiber protein expression. This suggested a crucial role of the five penultimate repeats in the crystallisation process. Chimeric pentons made of Ad5 penton base and fiber domains from different serotypes were analyzed with respect to crystal formation. No crystal was found when fiber consisted of shaft (S) from Ad5 and knob (K) from Ad3 (heterotypic S5-K3 fiber), but occurred with homotypic S3K3 fiber. However, less regular crystals were observed with homotypic S35-K35 fiber. TB5, a monoclonal antibody directed against the Ad5 fiber knob was found by immunofluorescence microscopy to react with high efficiency with the intranuclear protein crystals in situ. Data obtained with Ad fiber mutants indicated that the absence of crystalline inclusions correlated with a lower infectivity and/or lower yields of virus progeny, suggesting that the protein crystals might be involved in virion assembly. Thus, we propose that TB5 staining of Ad-infected 293 cells can be used as a prognostic assay for the viability and productivity of fiber-modified Ad5 vectors.\ud \ud

CONTEXT Delayed arterial hemorrhage is a rare complication after pancreaticoduodenectomy (frequency 2-4%) but carries a high mortality, with up to a third of patients dying as a consequence. Its ideal management remains unclear.Between 1993 and 2007, 317 head of pancreas resections were performed at our institution; there were 5 cases of delayed arterial hemorrhage (frequency 1.6%).This paper presents our experience in the management of delayed arterial haemorrhage post pancreaticoduodenecomy and discusses the role of angiograms, endoscopy and laparotomy in its management.

This paper highlights our experience of the transfer of hydrodynamic gene therapy (HGT) from the large animal, the pig, into clinical practice. The modification of balloon catheters and the development of a minimally invasive technique to allow selective isolation of liver segments for HGT in the large animal and human are described. Finally, our preliminary results from a phase I clinical study of HGT for thrombopoietin (TPO) in cirrhotic patients with thrombocytopenia are discussed. Based on these provisional data, minimally invasive selective HGT of liver segments appears to be technically safe, but further work is required to optimize the efficiency of gene transfer in order to achieve clinical benefit.

BACKGROUND: During pregnancy, fetal cells enter the maternal bloodstream resulting in fetal cell microchimerism. The fetal cells persist in the mother for decades and colonize a variety of maternal organs. They are associated with maternal autoimmune diseases and may also participate in tissue repair. The identity of the microchimeric cells is not certain but they must be able to persist long-term and have potential for multitissue differentiation. METHODS AND RESULTS: Here we tested the hypothesis that the fetal microchimeric cells are primitive stem cells, represented by CD34 + adherent cells, which have a wide potential for differentiation. We isolated these stem cells from the blood of pregnant females (n = 25) and detected fetal cells of the correct gender, using fluorescence in situ hybridization, in a high proportion (71% male fetuses and 90% female fetuses; false positive rate 11%, false negative rate 29%) of cases. By RT-PCR, we demonstrated that the cells express Oct-4, Nanog and Rex-1. No fetal cells were detected in the mononuclear or total CD34 + cell populations but high frequencies (mean 11.8 %) of fetal cells were detected in the adherent CD34+ cell population. CONCLUSIONS: These results identify adherent CD34+ stem cells as candidate fetal microchimeric cells, which are capable of sustaining the fetal cell population in the long term and have the ability to colonize multiple tissues and organs.

Advancement in radiological imaging technique has been crucial in the development of most of the surgical fields. Planning for operations needs good visualization of the organ and localization of the lesion within it, which was possible by Computed Tomography (CT) & Magnetic Resonance Imaging (MRI). However, without knowledge of major blood vessels or other important structures related to the lesion, surgery cannot be performed curatively and safely at the same time. In neurosurgery, this fact has been early recognized and three-dimensional (3D) imaging technique widely adopted to increase accuracy of the procedures. Similarly, reconstructed 3D images have been very useful in orthopaedic and maxillofacial surgery as well [1]

The gene therapy dream started well over a decade ago. Despite its wide and successful application in research, it still has not reached the clinics in a meaningful way. No doubt that successful gene therapy has a lot to offer to patients with inherited, benign or malignant diseases

Despite improvements in the management of liver tumours, both primary and metastatic, these tumours are still considered one of the most common malignancies worldwide with a high mortality rate and disappointing long-term survival rates. Hepatic resection has become the standard modality of treatment for patients with liver tumours, and currently remains the only potentially curative therapy. Many factors can affect hepatectomy, such as tumour size, location, multifocality, patients’ status, and hepatic function. But in order to attain a tumour free margin extensive hepatectomy is often necessary, which has led to an increased survival and improved outcome after hepatic resection for liver tumours: colorectal metastases [1, 2], hilar cholangiocarcinoma [3, 4] and hepatocellular carcinoma [5, 6, 7]

Those diseases which medicines do not cure, iron (the knife?) cures; those which iron cannot cure, fire cures; and those which fire cannot cure, are to be reckoned wholly incurable

Organ transplantation currently remains the definitive cure for several liver and pancreatic disorders. However, due to a major shortage of available organs many are turning to alternative sources for whole organ or cell-based transplantation. Stem cells have been proposed as an alternative to organ transplantation. The ethical and legal issues associated with embryonic stem cells have shifted the focus to adult stem cells and their regenerative potential has been under intense investigation. In this chapter we are reviewing the latest advances in stem cell therapy in liver diseases from bench to bedside.

Background Radiofrequency has been used as a tool for liver resection since 2002. A new laparoscopic device is reported in this article that assists liver resection laparoscopically. Methods From October 2006 to the present, patients suitable for liver resection were assessed carefully for laparoscopic resection with the laparoscopic Habib Sealer (LHS). Detailed data of patients resected laparoscopically with this device were collected prospectively and analyzed. Results In all, 28 patients underwent attempted laparoscopic liver resection. Four cases had to be converted to an open approach because of extensive adhesions from previous colonic operations. Twenty-four patients completed the procedure comprising tumorectomy (n = 7), multiple tumoretcomies (n = 5), segmentectomy (n = 3), and bisegmentectomies (n = 9). Vascular clamping of portal triads was not used. The mean resection time was 60 ± 23 min (mean ± SD), and blood loss was 48 ± 54 mL. None of the patients received any transfusion of blood or blood products perioperatively or postoperatively. Postoperatively, 1 patient developed severe exacerbation of asthma that required steroid therapy, and 1 other patient had a transient episode of liver failure that required supportive care. The mean duration of hospital stay was 5.6 ± 2 days (mean ± SD). At a short-term follow up, no recurrence was detected in patients with liver cancer. Conclusion Laparoscopic liver resection can be performed safely with this new laparoscopic liver resection device with a significantly low risk of intraoperative bleeding or postoperative complications.

Radiofrequency (RF)-assisted liver resection has been shown to allow virtually bloodless procedures without the need for vascular exclusion maneuvers. The aim of this study was to evaluate the safety and feasibility of RF-assisted liver resection in cirrhotic patients with hepatocellular carcinoma (HCC) and, moreover, to assess whether the RF-assisted procedure influenced the outcomes in terms of morbidity and mortality. This retrospective study included 39 cirrhotic patients who underwent RF-assisted liver resection for HCC between September 2001 and March 2006. In this period, we performed 17 monosegmentectomies, 16 bisegmentectomies, 4 trisegmentectomies, and 2 right hepatectomies. We never performed vascular exclusion maneuvers. Blood transfusion was necessary in 3 cases. One patient died postoperatively because of untreatable hepatorenal syndrome. The morbidity rate was 23%. Among patients without postoperative morbidity (n = 30), the 4-year survival rate was 61% versus 23% in patients who experienced postoperative morbidity (n = 9; P.05). The 4-year disease-free survival rate was 40% in patients without postoperative morbidity versus 10% in patients who experienced postoperative morbidity (P = .05). Tumor dimension, number of lesions, and Child-Pugh class at surgery did not seem to significantly influence the 4-year overall survival (P.05). In conclusion, RF-assisted liver resection was a safe, feasible procedure associated with low morbidity and hospital mortality rates even in cases of liver cirrhosis. It is, in our opinion, highly recommended for patients with HCC not suitable for liver transplantation.

Background: Liver surgery carries the risk of intraoperative bleeding. In order to avoid bleeding, transection of the liver can be performed after coagulating the parenchyma by using monoplolar or bipolar radiofrequency energy. Methods: 236 consecutive patients underwent liver resection with the radiofrequency-assisted technique using either a monopolar or a bipolar device. Data were collected prospectively to assess the outcome including, intraoperative blood loss, blood transfusion requirement, morbidity and mortality rates. Results: There were 41 major hepatectomies and 195 minor resections. Overall mean intraoperative blood loss was 157 ± 240 ml, while mean blood loss during liver transection was 90 ± 105 ml. 10 patients (4%) received blood transfusion. 50 patients (21%) developed postoperative complications including 5 bile leaks (2%). The mortality rate was 2.1%. No patient was reoperated for postoperative haemorrhage or bile leak. The mean postoperative stay was 11 ± 10 days. Conclusion: The radiofrequency-assisted liver resection technique offers hepatobiliary surgeons an additional method for performing liver resections with minimal blood loss, low transfusion requirement, and low morbidity and mortality rates.