DHPS-dependent hypusination of eIF5A1/2 is necessary for TGFß/fibronectin-induced breast cancer metastasis and associates with prognostically unfavorable genomic alterations in TP53.

Metastasis is the leading cause of mortality in patients with solid tumors. In this regard, we previously reported that Pseudopodium-Enriched Atypical Kinase One (PEAK1) is necessary for non-canonical Transforming Growth Factor ß (TGFß) signaling and TGFß/fibronectin-induced metastasis. Here, we demonstrate that inhibition of DHPS-dependent eIF5A1/2 hypusination blocks PEAK1 and E-Cadherin expression, breast cancer cell viability and TGFß/fibronectin-induced PEAK1-dependent breast cancer metastasis. Interestingly, TGFß stimulation of high-grade metastatic breast cancer cells increases and sustains eIF5A1/2 hypusination. We used a suite of bioinformatics platforms to search biochemical/functional interactions and clinical databases for additional control points in eIF5A1/2 and PEAK1-Epithelial to Mesenchymal Transition (EPE) pathways. This effort revealed that interacting EPE genes were enriched for TP53 transcriptional targets and were commonly co-amplified in breast cancer patients harboring inactivating TP53 mutations. Taken together, these results suggest that combinatorial therapies targeting DHPS and protein activities elevated in TP53-mutant breast cancers may reduce systemic tumor burden and improve patient outcomes.

MicroRNA-30a Inhibits Colorectal Cancer Metastasis Through Down-Regulation of Type I Insulin-Like Growth Factor Receptor.

BACKGROUND: miR-30a expression is down-regulated and regulates tumor suppressors in various cancers. AIM: We investigated the mechanisms underlying the biological role of miR-30a in CRC. METHODS: MicroRNA, mRNA, and protein expression were analyzed by quantitative real-time PCR and Western blot. The migration and invasion abilities of CRC were determined by wound healing assay, and trans-well migration and invasion. A luciferase reporter assay was used to confirm the targets of miR-30a. RESULTS: miR-30a expression was significantly down-regulated in CRC tissues and in CRC tissue with lymph node metastasis compared to CRC tissue without metastasis. Overexpression of miR-30a suppressed migration and invasion through insulin-like growth factor 1 receptor (IGF1R) in CRC cells. miR-30a suppresses IGF1R protein expression and inhibits ß-catenin or p-AKT and increases E-cadherin expression. The IGF1R expression level is also up-regulated in CRC tumors and inversely correlated with miR-30a in CRC specimens. CONCLUSIONS: miR-30a functions as a tumor-suppressive miRNA, which may provide a therapeutic strategy for metastasis of CRC.

MicroRNA-30a-5p (miR-30a) regulates cell motility and EMT by directly targeting oncogenic TM4SF1 in colorectal cancer.

PURPOSE: Colorectal cancer (CRC) is one of the leading causes of cancer death worldwide, and many oncogenes and tumor suppressor genes are involved in CRC. MicroRNAs (miRNAs) are small non-coding RNAs that can negatively regulate gene expression. Previous studies have revealed that miRNAs regulate the development and progression of many cancers. In this study, we investigated the role of microRNA-30a-5p (miR-30a) in CRC and its unknown mechanisms. METHODS: qRT-PCR was used to detect miR-30a and TM4SF1 mRNA expression in CRC specimens and cell lines. CRC cell migration and invasion were assessed after transfection with miR-30a or TM4SF1 using wound healing and trans-well migration and invasion assays. Transmembrane-4-L-six-family protein (TM4SF1) was validated as a target of miR-30a in CRC through luciferase reporter assay and bioinformatics algorithms. Moreover, two EMT regulators, E-cadherin and VEGF, were also identified using Western blotting and immunohistochemistry. RESULTS: We found that miR-30a was down-regulated in CRC tumor tissues and cell lines, and miR-30a was inversely associated with advanced stage and lymph node metastatic status compared with normal tissues. miR-30a decreased migration and invasion in CRC cell lines, and miR-30a overexpression not only down-regulated TM4SF1 mRNA and protein expression, but also inhibited the expression of VEGF and enhanced expression of E-cadherin. We also showed that TM4SF1 was up-regulated in CRC tumor specimens compared with adjacent normal tissues, and TM4SF1 expression was significantly associated with advanced stage and lymph node status compared with adjacent normal tissues. CONCLUSIONS: These results suggest that miR-30a is an important regulator of TM4SF1, VEGF, and E-cadherin for CRC lymph node metastasis, a potential new therapeutic target in CRC.

Discrimination Factors and Incorporation Rates for Organic Matrix in Shark Teeth Based on a Captive Feeding Study.

Sharks migrate annually over large distances and occupy a wide variety of habitats, complicating analysis of lifestyle and diet. A biogeochemical technique often used to reconstruct shark diet and environment preferences is stable isotope analysis, which is minimally invasive and integrates through time and space. There are previous studies that focus on isotopic analysis of shark soft tissues, but there are limited applications to shark teeth. However, shark teeth offer an advantage of multiple ecological snapshots and minimum invasiveness during removal because of their distinct conveyor belt tooth replacement system. In this study, we analyze δ13C and δ15N values of the organic matrix in leopard shark teeth (Triakis semifasciata) from a captive experiment and report discrimination factors as well as incorporation rates. We found differences in tooth discrimination factors for individuals fed different prey sources (mean ± SD; Δ13Csquid = 4.7‰ ± 0.5‰, Δ13Ctilapia = 3.1‰ ± 1.0‰, Δ15Nsquid = 2.0‰ ± 0.7‰, Δ15Ntilapia = 2.8‰ ± 0.6‰). In addition, these values differed from previously published discrimination factors for plasma, red blood cells, and muscle of the same leopard sharks. Incorporation rates of shark teeth were similar for carbon and nitrogen (mean ± SE; λC = 0.021 ± 0.009, λN = 0.024 ± 0.007) and comparable to those of plasma. We emphasize the difference in biological parameters on the basis of tissue substrate and diet items to interpret stable isotope data and apply our results to stable isotope values from blue shark (Prionace glauca) teeth to illustrate the importance of biological parameters to interpret the complex ecology of a migratory shark.

[(18)F] FP-CIT PET study in parkinsonian patients with leukoaraiosis.

BACKGROUNDS: Leukoaraiosis may present with slowly progressive parkinsonism indistinguishable from primary degenerative parkinsonism. Both leukoaraiosis and degenerative parkinsonism are an age-related disorder. Thus, comorbidity is expected to be common in elderly patients with parkinsonism. However, no systematic study has been reported on the clinical features indicating concomitant nigrostriatal dopaminergic denervation (NDD) in parkinsonian patients with leukoaraiosis. METHODS: We performed [(18)F] FP-CIT positron emission tomography studies in 42 consecutive parkinsonian patients with diffuse leukoaraiosis, but no basal ganglia vascular lesions. RESULTS: Twenty (48%) of the 42 patients had coexisting NDD. Compared to parkinsonian patients with isolated leukoaraiosis, those with coexisting NDD more frequently had asymmetric onset. They had similar degree of parkinsonian motor deficits in the legs, but greater rigidity and resting tremor in the arms. Consequently, they had less prominent lower body parkinsonism. They more frequently showed favorable response to levodopa treatment. They had similar burden of regional and total leukoaraiosis. Among a variety of clinical variables and MRI findings, only asymmetric onset and more than 30% improvement in UPDRS motor score by levodopa treatment were valuable indicators of coexisting NDD. CONCLUSIONS: We would like to recommend dopaminergic functional imaging studies for all parkinsonian patients with leukoaraiosis. Further studies are needed to confirm sensitivity and specificity of asymmetric onset and good levodopa response for the prediction of coexisting NDD in a different group of parkinsonian patients with leukoaraiosis.

Stenting from the vertebral artery to the posterior inferior cerebellar artery.

BACKGROUND AND PURPOSE: There are only a few reports on the feasibility and safety of stents used in the PICA, and clinical and angiographic follow-up results have not been fully addressed. We report our experiences of treating PICA origin or vertebral artery-PICA lesions by using self-expanding stents as adjuvant or rescue therapy with angiographic and clinical follow-up results. MATERIALS AND METHODS: Six patients were treated with self-expanding stent placements from the vertebral artery to the PICA. Two patients had a vertebral artery dissecting aneurysm involving the PICA origin, 3 had vertebral artery-PICA aneurysms, and 1 had segmental stenosis of the vertebral artery harboring the origin of the PICA. The safety, feasibility, and follow-up angiographic results were retrospectively evaluated. RESULTS: All procedures were successfully performed without any procedure-related complications. None of the patients showed PICA territorial infarction on DWI posttreatment. All patients were neurologically intact during the clinical follow-up of 3-24 months following the procedure. Follow-up angiography was performed at between 6 and 12 months in 5 of the 6 patients and was scheduled for the sixth patient but was not performed. The PICA showed good patency without in-stent stenosis in all 5 patients. CONCLUSIONS: In patients with lesions of the PICA origin or vertebral artery-PICA lesions, vertebral artery-to-PICA stent placement may be an option for preserving PICA patency in selected cases.

Benthic changes at McMurdo Station, Antarctica following local sewage treatment and regional iceberg-mediated productivity decline.

McMurdo Station, the largest research station in Antarctica, ceased on-site garbage dumping in 1988 and initiated sewage treatment in 2003. In 2003-2004 its sea-ice regime was altered by the massive B-15A and C-19 iceberg groundings in the Ross Sea, approximately 100km distant. Here we follow macrofaunal response to these changes relative to a baseline sampled since 1988. In the submarine garbage dump, surface contaminants levels have declined but associated macrofaunal recolonization is not yet evident. Although sewage-associated macrofauna were still abundant around the outfall nearly 2yr after initiation of treatment, small changes downcurrent as far as 434m from the outfall suggest some community recovery. Widespread community changes in 2003-2004, not seen in the decade previously, suggests that the benthos collectively responded to major changes in sea-ice regime and phytoplankton production caused by the iceberg groundings.

Chondrogenic differentiation of human umbilical cord blood-derived multilineage progenitor cells in atelocollagen.

BACKGROUND: For successful stem cell-based therapy, not only are alternative good cell sources needed but appropriate scaffolds are key factors. The main purpose of this study was to evaluate the multipotentiality of multilineage progenitor cells (MLPC) and assess the three-dimensional cultivation and chondrogenic differentiation of MLPC in atelocollagen gel for application of tissue-engineered cartilage constructs. METHODS: MLPC, human umbilical cord blood-derived clonal cell lines, from BioE Inc. were used. Immunophenotypes of MLPC were characterized using a fluorescence-activated cell sorter (FACS). In vitro differentiation potentials into osteogenic, adipogenic and chondrogenic lineages were examined. Differentiated cells were characterized by reverse transcriptase-polymerase chain reaction (RT-PCR), histologic and immunofluorescence staining. RESULTS Clonogenic MLPC maintained immunophenotypes with specific surface markers of mesenchymal stromal cells (MSC). The osteogenic and adipogenic potentials of MLPC were demonstrated by quantitative real-time PCR, alkaline phosphates activity and Oil Red O staining. Furthermore, MLPC were successfully differentiated into chondrocytes in atelocollagen gel, which was confirmed by RT-PCR and immunofluorescence staining for type II collagen protein. DISCUSSION: Whenever MSC are considered for the treatment of cartilage defects, a variety of scaffolds have been utilized as successful carriers for cell delivery. Our results suggest that MLPC can serve as an alternative source for stem cell-based therapy and transplantation. The chondrogenic potential of MLPC in atelocollagen could be suitable for cartilage tissue engineering.

Infantile head injury, with special reference to the development of chronic subdural hematoma.

An infantile head injury has unique features in that infants are totally helpless and dependent on their parents, and biomechanical characteristics of the skull and brain are very different from those of other age groups. The authors reviewed a total of 16 infant head injury patients under 12 months of age who were treated in our hospital from 1989 to 1997. Birth head injury was excluded. The most common age group was 3-5 months. Early seizures were noted in 7 cases, and motor weakness in 6. Three patients with acute intracranial hematoma and another 3 with depressed skull fracture were operated on soon after admission. Chronic subdural hematomas (SDHs) developed in 3 infants. Initial CT scans showed a small amount of SDH that needed no emergency operation. Resolution of the acute SDH and development of subdural hygroma appeared on follow-up CT scans within 2 weeks of injury. Two of these infants developed early seizures. Chronic SDH was diagnosed on the 68th and 111th days after the injuries were sustained, respectively. The third patient was the subject of close follow-up with special attention to the evolution of chronic SDH in view of our experience in the previous 2 cases, and was found to have developed chronic SDH on the 90th day after injury. All chronic SDH patients were successively treated by subduro-peritoneal shunting. In conclusion, the evolution of chronic SDH from acute SDH is relatively common following infantile head injury. Infants with head injuries, especially if they are associated with acute SDH and early development of subdural hygroma, should be carefully followed up with special attention to the possible development of chronic SDH

Comparison between of the attenuated BR-Oka and the wild type strain of Varicella Zoster Virus (VZV) on the DNA level.

Oka strain VR-795 (Varicella Zoster Virus, VZV) of American Type Culture Collection (ATCC) has been used for chickenpox vaccine production. In order to use this strain for vaccine production, the strain must be identified and its stability must be confirmed. The identification of the Oka strain has been confirmed using Restriction Fragment Length Polymorphism (RFLP) and DNA sequence analysis of glycoprotein-II (gp-II). The amino acid sequences of Oka deduced from the DNA sequence of gp-II have changed at three amino acids against Ellen and at one amino acid against Webster. To prove the stability of the Oka strain during the passage, RFLP and DNA sequence analyses were also used with 11, 15 and 23 times of virus passage. We found that the Oka strain was stable at passages of up to 23 times, based on the RFLP and DNA sequence analyses. The confirmed Oka strain was renamed as BR-Oka for the purposes of chickenpox vaccine production.

NSAIDs, aspirin, and esophageal strictures: are over-the-counter medications harmful to the esophagus?

There are several studies that suggest that aspirin (acetylsalicylic acid [ASA]) and nonsteroidal anti-inflammatory drugs (NSAIDs) are associated with esophagitis or esophageal stricture formation. There are limited data on the potential of low-dose ASA and over-the-counter (OTC) NSAIDs to cause esophageal injury. The goal of this study was to determine whether there is an association between esophageal strictures and ASA/NSAID use, including low-dose ASA and OTC NSAIDs. A total of 79 consecutive patients (mean age, 52.8 years; 38 men, 41 women) referred for endoscopy from 4/1/96 to 11/15/96 for chronic gastroesophageal reflux disease symptoms were evaluated. Data collected include gender, race, and age, NSAID or ASA use, as well as an assessment of dysphagia, heartburn duration, and heartburn frequency. Patients taking NSAIDs or ASA at least twice a week were considered ASA/NSAID users. There were 46 patients without strictures and 33 patients with peptic strictures. Patients with strictures were older than patients without strictures (mean age, 58.7 versus 48.6 years; p < 0.01), had longer duration of heartburn symptoms (8.6 versus 6.4 years, p < 0.05), and were more likely to have mucosal injury (50% versus 26.1%). Stricture patients were more likely to use ASA/NSAIDs (63.6% versus 26.1%; p < 0.01). In particular, stricture patients were more likely to use low-dose ASA than patients without strictures (30.3% versus 2.2%; p < 0.01). Otherwise, there were no significant differences with regard to gender, race, or heartburn duration or frequency. Linear regression analysis showed that ASA/NSAID use had a greater influence on the incidence of peptic strictures than age. There is an association between esophageal stricture and ASA/NSAID use, which includes OTC NSAIDs and low-dose ASA.

The prevalence of intestinal metaplasia in patients with and without peptic strictures.

OBJECTIVE: Several studies suggest that patients with esophageal peptic strictures have a high prevalence of Barrett's esophagus. However, these studies did not include appropriate control groups, were retrospective in nature, or did not strictly define Barrett's esophagus. Our aim was to compare the prevalence of Barrett's esophagus in patients with and without gastroesophageal reflux disease strictures in a prospective study. METHODS: Seventy-nine patients referred for endoscopy for gastroesophageal reflux disease symptoms were evaluated. We collected demographic information and an esophageal symptom assessment. Biopsy specimens were obtained from peptic strictures, Schatzki rings, or from any areas of columnar-lined esophagus or mucosal injury. Barrett's esophagus was strictly defined as the presence of intestinal metaplasia from tubular esophagus. RESULTS: There were 46 patients without strictures and 28 patients with peptic strictures. Five patients had Schatzki's rings. The prevalence of intestinal metaplasia was 23.9% in patients without strictures, and 25% in patients with peptic strictures (p = NS). There was no difference in prevalence of short- or long-segment Barrett's esophagus between the groups. Patients with strictures were older than patients without strictures (mean age 58.9 vs 48.6 yr), and more likely to have mucosal injury (50% vs 26.1%). Otherwise, there were no significant differences with regards to gender, race, heartburn duration or frequency. CONCLUSIONS: Barrett's esophagus, as defined by the presence of intestinal metaplasia in the tubular esophagus, is equally common in patients with and without peptic strictures. There does not appear to be an association between Barrett's esophagus and peptic strictures.

Chongmyungtang attenuates kainic acid-induced seizure and mortal effect in the mouse.

TheChongmyungtang (CMT; the combination ofAcorus gramineus, polygala tenuifolia andPoria cocos) has been recognized to possess the preventive effect against several neurologic disorders in human. In this study, we examined the effect of CMT on the three parameters associated with kainic acid (KA)-induced neurotoxicities; seizure/mortality, increased fos-related antigen (FRA) and glial fibrillary acidic protein (GFAP) expression. KA induced vigorous convulsions lasting 4-6 hr. Pretreatments with CMT before KA injection significantly reduced the seizure intensity as well as the mortality. CMT pretreatments also attenuated the KA-induced increase in FRA/GFAP expression in the hippocampus. These results suggest that CMT has a neuroprotective effect against KA-induced neurotoxicities.

In-111 labeled octreotide imaging of a primary carcinoid lesion undetected by conventional imaging studies in a patient with "chronic pancreatitis".

This is a case report of a patient with an initial diagnosis of chronic pancreatitis who actually had metastatic carcinoid tumor. His symptoms of abdominal pain, weight loss, and diarrhea were manifestations of the large tumor bulk within the liver as well as carcinoid syndrome. Although abdominal CT scans showed multiple liver lesions, the primary lesion was not identified by conventional imaging studies. However, the mid-gut primary lesion was visualized on in-111 labeled octreotide scintigraphy; where the liver lesions were better delineated and seen to be separate from the normal pancreas when the Tc-99m sulfur colloid images were compared to the octreotide images.

Diagnostic inconsistencies in Barrett's esophagus. Department of Veterans Affairs Gastroesophageal Reflux Study Group.

BACKGROUND/AIMS: Few studies have compared the precision of various diagnostic tests used to determine the presence of Barrett's esophagus. The aim of this study was to compare the results of histological, endoscopic, and manometric tests for patients with Barrett's esophagus in two closely spaced examinations. METHODS: In a Veterans Administration Cooperative Study, 192 patients with complicated gastroesophageal reflux disease had esophageal manometry and endoscopy performed at baseline and after 6 weeks. At each examination, the endoscopist localized the most proximal level of Barrett's epithelium and the lower esophageal sphincter and obtained esophageal biopsy specimens. RESULTS: One hundred sixteen patients met the criteria for Barrett's esophagus on at least one of the two endoscopic examinations. Among patients with specialized columnar epithelium, 20% had specialized columnar epithelium found on only one of the two examinations. Although the mean lower esophageal sphincter level did not change, approximately 10% of patients had a change > or = 4 cm on endoscopy and manometry between examinations. This led to an apparent change in the diagnosis in 18% of patients with Barrett's esophagus. CONCLUSIONS: From one endoscopic examination to another, inconsistencies in the ability to detect specialized columnar epithelium are common. This may lead to substantial problems in establishing an accurate diagnosis of Barrett's esophagus.

Crystallization of phosphatidylinositol-specific phospholipase C from Bacillus cereus.

Phosphatidylinositol-specific phospholipase C (PI-PLC) cleaves phosphoinositides into two parts, lipid-soluble diacylglycerol and the water-soluble phosphorylated inositol. Two crystal forms of Bacillus cereus PI-PLC have been obtained by the vapor diffusion technique. Hexagonal crystals were grown from solutions containing polyethylene glycol (PEG; 4,000 to 8,000 D). The space group of these hexagonal crystals is P6(1)22 (or the enantiomorphic space group P6(5)22), with cell constants a = b = 133 A, and c = 231 A. The crystals diffract to 2.8 A. The second crystalline form was grown from a two-phase PEG (600 D)-sodium citrate solution. The phase diagram and PI-PLC distribution between phases has been determined. The enzyme crystallizes from the PEG-rich phase. The crystals are orthorhombic with space group P2(1)2(1)2(1) (a = 45 A, b = 46 A, c = 160 A), and contain one PI-PLC monomer per asymmetric unit. The orthorhombic crystals diffract to 2.5 A. Both the hexagonal and orthorhombic forms are suitable for crystallographic studies.