Isolation of a new antibacterial peptide actinokineosin from Actinokineospora spheciospongiae based on genome mining.

Based on genome mining, a new antibacterial peptide named actinokineosin was isolated from a rare actinomycete Actinokineospora spheciospongiae. The amino acid sequence of the C-terminus of actinokineosin was established by TOF-MS/MS experiments. The amino acid sequence in the macrolactam ring was determined by TOF-MS/MS analyses after cleavage with BNPS-skatole and successive trypsin treatment. As a result of an antibacterial assay using a paper disk, actinokineosin showed antibacterial activity against Micrococcus luteus at a dosage of 50 µg per disk. From the genome sequence data of A. spheciospongiae, the biosynthetic gene cluster of actinokineosin was found and was indicated to consist of 10 genes. Among the genes, the gene aknA encoded the precursor of actinokineosin and the genes including aknC, aknB1 and aknB2 were proposed as modification enzymes to give mature actinokineosin. SIGNIFICANCE AND IMPACT OF THE STUDY: Genome mining is a powerful tool to find new bioactive compounds from the genome database. In this report, we succeeded in isolation and structure determination of a new antibacterial peptide named actinokineosin based on genome mining.

Evidence for biological nitrification inhibition in Brachiaria pastures.

Nitrification, a key process in the global nitrogen cycle that generates nitrate through microbial activity, may enhance losses of fertilizer nitrogen by leaching and denitrification. Certain plants can suppress soil-nitrification by releasing inhibitors from roots, a phenomenon termed biological nitrification inhibition (BNI). Here, we report the discovery of an effective nitrification inhibitor in the root-exudates of the tropical forage grass Brachiaria humidicola (Rendle) Schweick. Named "brachialactone," this inhibitor is a recently discovered cyclic diterpene with a unique 5-8-5-membered ring system and a gamma-lactone ring. It contributed 60-90% of the inhibitory activity released from the roots of this tropical grass. Unlike nitrapyrin (a synthetic nitrification inhibitor), which affects only the ammonia monooxygenase (AMO) pathway, brachialactone appears to block both AMO and hydroxylamine oxidoreductase enzymatic pathways in Nitrosomonas. Release of this inhibitor is a regulated plant function, triggered and sustained by the availability of ammonium (NH(4)(+)) in the root environment. Brachialactone release is restricted to those roots that are directly exposed to NH(4)(+). Within 3 years of establishment, Brachiaria pastures have suppressed soil nitrifier populations (determined as amoA genes; ammonia-oxidizing bacteria and ammonia-oxidizing archaea), along with nitrification and nitrous oxide emissions. These findings provide direct evidence for the existence and active regulation of a nitrification inhibitor (or inhibitors) release from tropical pasture root systems. Exploiting the BNI function could become a powerful strategy toward the development of low-nitrifying agronomic systems, benefiting both agriculture and the environment.

Minimum stable structure of the receptor for advanced glycation end product possesses multi ligand binding ability.

The receptor for advanced glycation end products (RAGE) is a multi-ligand receptor involved in the development of diabetic complications. Although the soluble form of the extracellular domain maintains the ability to bind multi-ligands, it is unstable and degrades into several peptide species during storage. Proteolysis with thrombin or factor Xa revealed several protease sensitive sites. Most sensitive site is located between Arg228 and Val229, and peptide bond next to Arg216, Arg116, Arg114 and Trp271 are also cleaved. Seven truncated extracellular domains of RAGE were engineered in order to obtain a stable soluble fragment. RAGE 143 (Ala23-Thr143) is not only protease resistant but also shows the same ligand-binding ability as that of the full-length extracellular domain. The resultant minimum RAGE 143 works as a stable recognition devise to detect advanced glycation end products (AGEs).

Differential expression of the skeletal muscle proteome in grazed cattle.

The objective of this study was to investigate the differences in the muscle proteome of grass-fed and grain-fed cattle. Eight Japanese Black Cattle 10 mo of age were separated randomly into 2 groups: 1) grazing (grass-fed) and 2) concentrate (grain-fed) groups. All cattle were first housed individually in a stall barn and fed a combination of concentrate ad libitum and Italian ryegrass hay until 21 mo of age. After this control period, the 4 grass-fed cattle were placed on outdoor pasture, whereas the other 4 grain-fed cattle continued on the concentrate diet. The cattle were slaughtered at 27 mo of age, and tissues from the semitendinosus muscle were obtained for use in proteome analysis. Differential expression of muscle proteins in the 2 groups was carried out using 2-dimensional gel electrophoresis (2DE) and Western blot analyses, with subsequent mass spectrometry. Approximately 200 individual protein spots were detected and compared in each group using 2DE, of which 20 and 9 spots, respectively, showed differences in the spot intensity for the sarcoplasmic fraction and myofibrillar fraction. In the grazing group, the relative intensity of spots was significantly greater for adenylate kinase 1 and myoglobin in the sarcoplasmic fraction, and for slow-twitch myosin light chain 2 in the myofibrillar fraction (P < 0.05), than the concentrate group. The relative spot intensity of several glycolytic enzymes was significantly greater in the grazing group, such as beta-enolase 3, fructose-1,6-bisphosphate aldolase A, triosephosphate isomerase, and heat shock 27 kDa protein (P < 0.05). Moreover, significantly greater slow twitch of troponin T, troponin I, and myosin heavy chain of semitendinosus muscle was detected in the grazing group than in the concentrate group using Western blot analysis (P < 0.05). Several previous reports have described that the slow-twitch muscle contents affect elements of nutrition, flavor, and food texture of meat. This study revealed muscle fiber type conversion to slow-twitch tissues from fast-twitch tissues occurring with change in the energy metabolic enzyme when cattle were grazed in the latter fattening period. Although analyses of the influence on elements of nutrition, flavor, and food texture were not done for this study, these results show that slow-twitch converted muscle resulting from the grazing of cattle might modify several meat characteristics.

Clinical results of intra-arterial adjuvant chemotherapy for prevention of liver metastasis following curative resection of biliary tract cancer.

BACKGROUND: This is a report on the clinical results of intra-arterial adjuvant chemotherapy in the prevention of liver metastasis following curative resection of biliary tract cancer. METHODS: Nineteen patients of advanced biliary tract underwent a pathologically radical operation between 2001 and 2006 (8 M and 11 F; mean age 66.2 years). Intra-arterial adjuvant chemotherapy with CDDP and 5-FU was performed selectively for 9 patients. The control group comprised 10 patients. Age, gender, staging of the disease, resection of the portal vein, postoperative radiotherapy, histological liver invasion as demographics and clinical characteristics were compared between the two groups. RESULTS: Demographics and clinical characteristics were similar in the two groups. Liver metastasis occurred in 4 of 9 patients (44.4%) in the chemotherapy group and in 5 of 10 patients (50%) in the control group. There was no difference in the rate of liver metastasis between the two groups. The median survival term was 23.3 months for 9 patients who underwent the intra-arterial adjuvant chemotherapy, whereas the median survival term for 10 patients who were curatively resected without intra-arterial adjuvant chemotherapy was 21.7 months. The median survival term was statistically similar in both groups. Furthermore, in the recurrence-free survival, there was no major difference between the chemotherapy and control groups statistically. CONCLUSIONS: In the patients with advanced biliary tract cancer who underwent the curative operation, the intra-arterial adjuvant chemotherapy could not suppress the rate of liver metastasis nor improve cumulative survival.

Duct-to-mucosa pancreaticojejunostomies with a hard pancreas and dilated pancreatic duct and duct-to-mucosa pancreaticojejunostomies with a soft pancreas and non-dilated duct.

BACKGROUND: We performed duct-to-mucosa pancreaticojejunostomy with resection of jejunal serosa in 55 patients, and here compare the clinical results between duct-to-mucosa pancreaticojejunostomies with a non-dilated pancreatic duct and those with a dilated duct. PATIENTS AND METHODS: In the period 1999 to 2005, 55 patients (27 F, 28 M; mean age 63.4 years) underwent duct-to-mucosa pancreaticojejunostomy with resection of jejunal serosa. A non-dilated pancreatic duct was observe in 29 patients in group A and a dilated pancreatic duct in 26 patients in group B. Clinical characteristics (age, gender, benign or malignant condition, presence of diabetes mellitus, anastomotic time) were analyzed in both groups and postoperative complications were compared between groups. RESULTS: In a comparison of clinical characteristics, all factors were similar between groups. In group A, the postoperative complication occurred in 4 (wound infection in 2, pulmonary embolism in 1, gastric ulcer in 1) of 29 patients (13.8%), and in group B in 1 (pneumothorax) of 26 patients (3.8%). No pancreatic leakage was observed in either group. The difference between group A and group B in the rate of postoperative complication was not statistically significant. CONCLUSIONS: There was no statistical difference in the rate of postoperative complications, including pancreatic leakage, between duct-to-mucosa pancreaticojejunostomies with a dilated pancreatic duct and those with a non-dilated duct. We consider that the diameter of the pancreatic duct is irrelevant to results of duct-to-mucosa pancreaticojejunostomy.

Verrucotoxin, a stonefish venom, modulates calcium channel activity in guinea-pig ventricular myocytes.

BACKGROUND AND PURPOSE: Stonefish (Synanceia genus) are commonly found in shallow waters of the Pacific and Indian Oceans. The venom of stonefish is stored in the dorsal fine spines and contains a proteinaceous toxin, verrucotoxin (VTX). The stings produced by the spines induce intense pain, respiratory weakness, damage to the cardiovascular system, convulsions and paralysis, sometimes leading to death. Although there are many studies on VTX, the mechanism(s) underlying the VTX-mediated cardiotoxicity is not yet fully understood. The aim of this study was to investigate the modulation of ion channels in cardiac tissue by VTX. EXPERIMENTAL APPROACH: The effects of VTX on changes in the voltage or current in guinea-pig ventricular myocytes were investigated using a patch clamp method. KEY RESULTS: VTX (10 microg ml(-1)) prolonged the action potential duration by 2.5-fold. VTX increased L-type Ca(2+) currents (I (Ca(L))) in a concentration-dependent manner with a EC(50) value of 7 microg ml(-1) and a maximum increase of 3.1-fold. The non-selective beta-adrenoceptor antagonist, propranolol (1 microM) and the selective beta(1)-adrenoceptor antagonist, CGP20712A (10 microM) each abolished the effect of VTX (100 microg ml(-1)) on I (Ca(L)). Furthermore, the protein kinase A (PKA) antagonists H-89 (10 microM) and Rp-8-Br-cAMPS (30 microM) inhibited the effect of VTX on I (Ca(L)). CONCLUSIONS AND IMPLICATIONS: VTX modulates Ca(2+) channel activity through the beta-adrenoceptor-cAMP-PKA pathway.

Cerebral blood volume changes in patients with eating disorders during word fluency: a preliminary study using multi-channel near infrared spectroscopy.

OBJECTIVE: This study investigated the characteristics of cerebral oxygenation changes in eating disorders patients (ED) and normal controls during the cognitive tasks, using a highly time-resolved, and non-invasive instrument. METHOD: Eleven female patients with anorexia nervosa or bulimia nervosa were recruited, and 11 healthy females participated. The relative concentrations of oxy-hemoglobin [o-Hb] and deoxy-hemoglobin [d-Hb] were measured during word fluency task using multichannel near infrared spectroscopy (NIRS). RESULTS: The increases of o-Hb and d-Hb during the task were compared between the groups. ED patients showed lower activation and a gradual increase in o-HB during the task. In the frontal, d-HB concentrations decreased during the task in ED patients. CONCLUSION: These specific patterns of oxygenation changes may indicate less supply and less demand of cerebral blood volume. Bedside measurements of cerebral oxygenation changes using NIRS are useful on understanding of neurophysiological features of ED.

Muscle type specific expression of tropomyosin isoforms in bovine skeletal muscles.

Nucleotide sequences encoding an entire coding region for bovine tropomyosin (TPM) isoforms were determined. Three TPM isoforms, TPM1, TPM2 and TPM3, were expressed in bovine skeletal muscles, and exhibited a 93.3%, 99.6% and 100% amino acid homology to the human sequence, respectively. Based on the sequences, the composition of TPM isoforms was analyzed on cDNA and protein levels from five physiologically different muscles (masseter, diaphragm, psoas major, longissimus thoracis and semitendinosus) using RT-PCR and proteome analyses. Although the content of TPM2 was constantly about 50% of the total TPM in all muscles, the contents of TPM1 and TPM3 were different in muscles according to their function in muscle contraction. In masseter, the content of TPM3 cDNA was about 50% and higher than that of other muscles. In longissimus thoracis and semitendinosus, the contents of TPM1 cDNA were 29.6% and 31.7%, respectively, which were comparatively higher than that of other muscles. The result suggests that the TPM dimer consists of the TPM2 subunit regularly and TPM1 or TPM3 depending on whether the muscle is fast or slow type, respectively.

Ergosterol peroxide from an edible mushroom suppresses inflammatory responses in RAW264.7 macrophages and growth of HT29 colon adenocarcinoma cells.

BACKGROUND AND PURPOSE: 5alpha,8alpha-Epidioxy-22E-ergosta-6, 22-dien-3beta-ol (ergosterol peroxide) is a major antitumour sterol produced by edible or medicinal mushrooms. However, its molecular mechanism of action has yet to be determined. Here, we examine the anticancer and anti-inflammatory effects of ergosterol peroxide. EXPERIMENTAL APPROACH: After treating RAW264.7 macrophages with LPS and purified ergosterol peroxide or ergosterol, we determined LPS-induced inflammatory cytokines, nuclear DNA binding activity of transcription factors and phosphorylation of MAP kinases (MAPKs). HT29 colorectal adenocarcinoma cells were treated with ergosterol peroxide for 5 days. To investigate the antitumour properties of ergosterol peroxide, we performed DNA microarray and RT-PCR analyses and determined the reactive oxygen species (ROS) in HT29 cells. KEY RESULTS: Ergosterol peroxide suppressed LPS-induced TNF-alpha secretion and IL-1alpha/beta expression in RAW264.7 cells. Ergosterol peroxide and ergosterol suppressed LPS-induced DNA binding activity of NF-kappaB and C/EBPbeta, and inhibited the phosphorylation of p38, JNK and ERK MAPKs. Ergosterol peroxide down-regulated the expression of low-density lipoprotein receptor (LDLR) regulated by C/EBP, and HMG-CoA reductase (HMGCR) in RAW264.7 cells. In addition, ergosterol peroxide showed cytostatic effects on HT29 cells and increased intracellular ROS. Furthermore, ergosterol peroxide induced the expression of oxidative stress-inducible genes, and the cyclin-dependent kinase inhibitor CDKN1A, and suppressed STAT1 and interferon-inducible genes. CONCLUSION AND IMPLICATION: Our results suggest that ergosterol peroxide and ergosterol suppress LPS-induced inflammatory responses through inhibition of NF-kappaB and C/EBPbeta transcriptional activity, and phosphorylation of MAPKs. Moreover, ergosterol peroxide appears to suppress cell growth and STAT1 mediated inflammatory responses by altering the redox state in HT29 cells.

IgM-containing fraction suppressed voltage-gated potassium channels in acquired neuromyotonia.

OBJECTIVES: Acquired neuromyotonia (ANM) is an autoimmune disorder caused by antibodies to voltage-gated potassium channels (VGKC). Previously, we reported a patient with immunoglobulin M (IgM), instead of immunoglobulin G (IgG), anti-VGKC antibody. The purpose of this study was to determine the function of IgM-containing fraction in ANM patients. MATERIALS AND METHODS: We determined whether anti-VGKC antibodies in the IgG or IgM-containing fractions suppressed outward potassium current (OKC) using the patch clamp method in three patients with ANM. Whole sera from all patients suppressed OKCs. RESULT: Only the purified IgG, not the IgM-containing fractions from two patients suppressed VGKCs, whereas in a patient with IgM anti-VGKC antibody, only the IgM-containing fractions, not the IgG-containing fractions suppressed VGKCs. CONCLUSION: Anti-VGKC antibodies belonging to the IgM subclass should be determined in seronegative ANM patients.

Regional cerebral blood flow in Parkinson disease with nonpsychotic visual hallucinations.

BACKGROUND: Patients with Parkinson disease (PD) often experience visual hallucinations (VH) with retained insight (nonpsychotic) but the precise mechanism remains unclear. OBJECTIVE: To clarify which neural substrates participate in nonpsychotic VH in PD, the authors evaluated regional cerebral blood flow (rCBF) changes in patients with PD and VH. METHODS: The authors compared 24 patients with PD who had nonpsychotic VH (hallucinators) and 41 patients with PD who had never experienced VH (non-hallucinators) using SPECT images with N-isopropyl-p-[(123)I]iodoamphetamine. There were no significant differences in age, sex, duration of disease, doses of PD medications, Hoehn and Yahr scale, or Mini-Mental State Examination (MMSE) scores between the two groups. The rCBF data were analyzed using statistical parametric mapping (SPM). RESULTS: The rCBF in the right fusiform gyrus was lower in the hallucinators than in the non-hallucinators (corrected p < 0.05 at cluster levels). The hallucinators revealed higher rCBF in the right superior and middle temporal gyri than the non-hallucinators (uncorrected p < 0.001). These significant differences were demonstrated after MMSE scores and duration of disease, which are the relevant factors associated with VH, were covariated out. CONCLUSIONS: Nonpsychotic visual hallucinations in Parkinson disease (PD) may be associated with hypoperfusion in the right fusiform gyrus and hyperperfusion in the right superior and middle temporal gyri. These temporal regions are important for visual object recognition and these regional cerebral blood flow changes are associated with inappropriate visual processing and are responsible for nonpsychotic visual hallucinations in PD.

Three-dimensional stereotactic surface projection study of orthostatic hypotension and brain perfusion image in Parkinson's disease.

OBJECTIVE: To compare brain perfusion image using three-dimensional stereotactic surface projection (3D-SSP) analysis of N-isopropyl-p-123I iodoamphetamine (123I-IMP) single photon emission computed tomography (SPECT) between patients with Parkinson's disease with orthostatic hypotension and those without orthostatic hypotension. MATERIALS AND METHODS: Fifteen patients with Parkinson's disease and orthostatic hypotension and 13 patients with Parkinson's disease without orthostatic hypotension were studied. We compared brain perfusion image between the two groups by 3D-SSP. RESULTS: Bilateral anterior cingulate gyrus perfusion of the patients with orthostatic hypotension was significantly decreased compared to that of the patients without orthostatic hypotension. CONCLUSIONS: The disorder of anterior cingulate gyrus may participate in the autonomic failure in Parkinson's disease.

Analysis of acrylamide by LC-MS/MS and GC-MS in processed Japanese foods.

Acrylamide concentrations in processed foods (63 samples covering 31 product types) from Japan were analysed by LC-MS/MS and GC-MS methods. The limit of detection and limit of quantification of acrylamide were 0.2 ng x ml(-1) (6 fmol) and 0.8 ng x ml(-1) (22 fmol), respectively, by LC-MS/MS, and those of 2,3-dibromopropionamide derived from acrylamide were 12 ng x ml(-1) (52 fmol) and 40 ng x ml(-1) (170 fmol), respectively, by GC-MS. Repeatability given as RSD was <5 and <15% for the LC-MS/MS and GC-MS methods, respectively. High correlation (r(2) - 0.946) was observed between values obtained by the two methods. Most potato crisps and whole potato-based fried snacks showed acrylamide concentrations >1000 microg x kg(-1). The concentrations in non-whole potato-based snacks, rice crackers processed by grilling or frying, and candied sweet potatoes were lower compared with those in the potato crisps and the whole potato-based fried snacks. One of the whole potato-based fried snacks, however, showed low acrylamide concentration (<50 microg x kg(-1)) suggesting the formation of acrylamide is strongly influenced by processing conditions. Acrylamide concentrations in instant precooked noodles and won-tons were <100 microg x kg(-1) with only one exception. Roasted barley grains for 'Mugi-cha' tea contained 200-600 microg x kg(-1) acrylamide.

A case of idiopathic spinal epidural lipomatosis presented with radicular pain caused by compression with enlarged veins surrounding nerve roots.

OBJECTIVE: To report a case of idiopathic spinal epidural lipomatosis (SEDL) presented with unique radicular pain most likely caused by enlarged veins surrounding nerve roots. PATIENT: A 26-year-old male presented with radicular pain of the right T6-T7 area. He also showed Becker's nevus in the corresponding area. CT myelography and magnetic resonance imaging revealed epidural lipomatosis posterior to T4-T8 of the spinal cord. Surgical removal of adipose tissue and a hemilaminectomy of T4-T7 were performed and resulted in relief of the radicular pain. CONCLUSIONS: Lipomatosis was histologically confirmed and surrounded by enlarged veins. These abnormally enlarged veins compressed the nerve roots and were thought to cause radicular pain. Also, Becker's nevus of this case seems to have some relationship with SEDL.

Highly polymerized procyanidins in brown soybean seed coat with a high radical-scavenging activity.

1,1-diphenyl-2-picrylhydrazyl (DPPH) radical-scavenging activity of the 70% aqueous acetone extract from the seed coat of the brown soybean variety, Akita-Zairai, was investigated. The activity of the seed coat of Akita-Zairai was much higher than that of three other reddish-brown varieties, but lower than that of two black varieties, and was closely dependent on the content of phenolic compounds. In the LH20 column chromatography of Akita-Zairai, high DPPH radical-scavenging activities were detected in the fractions eluted with MeOH and 70% aqueous acetone. Proanthocyanidins were also detected in fractions showing high radical-scavenging activities. Matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry analysis showed that the degree of polymerization (DP) of the procyanidins contained in the brown or black soybean seed coat was as high as DP30.

Genome-wide screening of genes showing altered expression in liver metastases of human colorectal cancers by cDNA microarray.

In spite of intensive and increasingly successful attempts to determine the multiple steps involved in colorectal carcinogenesis, the mechanisms responsible for metastasis of colorectal tumors to the liver remain to be clarified. To identify genes that are candidates for involvement in the metastatic process, we analyzed genome-wide expression profiles of 10 primary colorectal cancers and their corresponding metastatic lesions by means of a cDNA microarray consisting of 9121 human genes. This analysis identified 40 genes whose expression was commonly upregulated in metastatic lesions, and 7 that were commonly downregulated. The upregulated genes encoded proteins involved in cell adhesion, or remodeling of the actin cytoskeleton. Investigation of the functions of more of the altered genes should improve our understanding of metastasis and may identify diagnostic markers and/or novel molecular targets for prevention or therapy of metastatic lesions.

[Role of radiology in forensic dentistry]. / O papel da Radiologia em Odontologia Legal.

There are various methods for the identification of human remains; most of them are based on comparisons between available ante- and post-mortem data. Although fingerprinting is the most accurate and precise method, in many cases, such as in mutilated, decomposed, burned or fragmented bodies, it cannot be used, and the dental methods become of the utmost importance, since teeth and dental restorations are very resistant to destruction by fire--they keep numerous characteristics, which are quite unique, and offer the possibility of accurate and legally acceptable identification of remains. This brief review describes the progress of forensic dentistry during the last 25 years, regarding the procedures and techniques that use ante-mortem and post-mortem radiographs. Among the discussed progresses one can point out: a specially designed self-supporting film holder that retains intra-oral films in the mouths of deceased persons whose mandibular musculature has been fixed in rigor mortis; positioning devices suitable for reproducing the geometry of ante-mortem radiographic images; methods involving digital radiographs, which can be easily stored in a central archive, retrieved and transmitted, via modem, to mass casualty sites; age estimation procedures; identification of edentulous individuals comparing radiographs of the maxilla; and studies of validation of dental radiographs taken with ante-mortem and post-mortem intervals of up to 30 years.

Does pleural lavage cytology before thoracic closure predict both patient's prognosis and site of cancer recurrence after resection of esophageal cancer?

BACKGROUND: Operative manipulation occasionally exfoliates and spreads cancer cells in the surgical field, and it is a matter of concern whether the exfoliated cancer cells actually affect the patient's prognosis and sites of cancer recurrence. METHODS: In 240 patients with esophageal cancers, lavage cytology (LC) of the right pleural cavity was performed before and after esophageal resection combined with regional lymphadenectomy. The cytologic results were compared with the pathologic factors associated with cancer extension, postoperative survival, and cause of surgical failure. RESULTS: Only 3 patients (1.3%) were LC positive before resection. Of the 237 LC-negative patients, LC was also negative after resection in 215 patients (90.7%) (LC-/-), but LC became positive after resection in 22 patients (9.3%) (LC-/+). The 3-year survival rate was 0% in the LC-/+ group versus 65% in the LC-/- group, and the median survival rates were 10.9 months and 25.0 months, respectively (P <.0001). Multivariate analysis revealed that LC-/+ was an independent prognostic factor (P =.0331), along with nodal involvement and depth of cancer invasion. However, there were no significant differences in the sites of cancer recurrence between the 2 groups. Only 1 patient was found to develop the first recurrence in the pleural cavity. The LC-/+ group had a higher incidence of bulky lymph-node metastasis (P =.0009). CONCLUSIONS: Pleural LC after resection of esophageal cancer seems to be a prognostic indicator of overall recurrence, but not necessarily in the pleural cavity. Patients with a positive LC after resection may benefit most by effective systemic adjuvant chemotherapy.

Reduced expression of focal adhesion kinase in liver metastases compared with matched primary human colorectal adenocarcinomas.

The focal adhesion kinase (FAK) is implicated in integrin-mediated signal transduction pathways used in cell adhesion, cell motility, apoptosis, and anchorage-independent growth. Because cancer invasion and metastasis are thought to be associated with alterations in cellular adhesive and motile properties, we studied the expression of four focal adhesion proteins including FAK in matched samples of human normal colorectal mucosa (N), primary colorectal adenocarcinomas (T) and liver metastases (M) from 10 patients by Western blot analysis. This gave us the advantage of directly comparing levels of focal adhesion protein expression within the same genetic background. Average FAK expression level was significantly higher in T than in N and it was significantly lower in M than in T. Average paxillin expression level was also significantly higher in T than in N, but it was not significantly different between T and M. Similar results were obtained by immunohistochemical analyses of FAK and paxillin expression. Average vinculin and talin expression levels showed no significant differences among these three samples (N, T, and M). These data demonstrate that the FAK expression level increases in primary tumors compared with normal mucosa and decreases in liver metastases to the level of normal mucosa in the majority of human colorectal adenocarcinomas. Up- and down-regulation of FAK protein expression observed in this study may have a profound effect on the signal transduction.