bluuebunny/crossref_metadata_2025 huggingface dataset:
Electrochemical Studies of Fluorenone in Ionic Liquids and Aprotic Solvents
Authors: Canby Dana , Cheek Graham T.
Published: October 2007

The electrochemical behavior of 9-fluorenone in acetonitrile / tetraethylammonium tetrafluoroborate (TEA BF4) solutions has been explored by additions of 1-ethyl-3-methylimidazolium tetrafluoroborate (EMIm BF4) and the proton donor 1,1,1,3,3,3- hexafluoro-2-propanol (HFIP). The effect of EMIm BF4 addition was found to be consistent with complexation of the dianion of 9-fluorenone by the EMIm cation, causing a positive shift of the second reduction process. Addition of HFIP as proton donor, on the other hand, caused the formation of a single reduction process near the first fluorenone reduction process.
Increased stiffness of the rat liver precedes matrix deposition: implications for fibrosis
Authors: Georges Penelope C. , Hui Jia-Ji , Gombos Zoltan , McCormick Margaret E. , Wang Andrew Y. , Uemura Masayuki , Mick Rosemarie , Janmey Paul A. , Furth Emma E. , Wells Rebecca G.
Published: October 2007

Liver fibrosis, the response to chronic liver injury, results from the activation of mesenchymal cells to fibrogenic myofibroblasts. We have recently shown that two key myofibroblast precursor populations, hepatic stellate cells and portal fibroblasts, undergo activation in culture in response to increasing substrate stiffness. We therefore hypothesized that alterations in liver stiffness precede myofibroblast activation and fibrosis in vivo as well. To test this hypothesis, we induced fibrosis in rats by twice weekly injections of carbon tetrachloride (CCl4) and then killed the animals at various time points ranging from 3 to 70 days after the initiation of injury. The shear storage modulus of the whole liver was measured on fresh tissue; fixed and frozen tissue from the same livers was used to quantify fibrosis. We observed that liver stiffness increased immediately and continued to increase, leveling out by day 28. Fibrosis, measured histologically by trichrome staining as well as by quantitative sirius red staining, increased with time, although these increases were delayed relative to changes in stiffness. There was no direct correlation between stiffness and fibrosis at early or late time points. Treatment of a second cohort of rats with the lysyl oxidase inhibitor, β-aminopropionitrile (BAPN), partially prevented early increases in liver stiffness. We concluded that increases in liver stiffness precede fibrosis and potentially myofibroblast activation. Liver stiffness appears to result from matrix cross-linking and possibly other unknown variables in addition to matrix quantity. We suggest that increased liver stiffness may play an important role in initiating the early stages of fibrosis.
The proinflammatory phenotype of PECAM-1-deficient mice results in atherogenic diet-induced steatohepatitis
Authors: Goel Reema , Boylan Brian , Gruman Lynn , Newman Peter J. , North Paula E. , Newman Debra K.
Published: October 2007

The severity of nonalcoholic steatohepatitis (NASH) is determined by environmental and genetic factors, the latter of which are incompletely characterized. Platelet endothelial cell adhesion molecule-1 (PECAM-1) is a 130-kDa transmembrane glycoprotein expressed on blood and vascular cells. In the present study, we provide data for the novel finding that genetic deficiency of PECAM-1 potentiates the development and progression of NASH. We found that the rate of development and severity of diet-induced NASH are markedly enhanced in PECAM-1-deficient [knockout (KO)] mice relative to wild-type (WT) mice, as measured by histological and biochemical evaluation. Livers from KO mice exhibited typical histological features of NASH, including macrovesicular fat accumulation, hepatocyte injury with infiltration of inflammatory cells, fibrosis, and heightened oxidative stress. Alanine aminotransferase, a marker for liver injury, was also significantly higher in KO compared with WT mice. Consistent with a role for PECAM-1 as a suppressor of proinflammatory cytokines, plasma levels of inflammatory cytokines, including TNF-α and monocyte chemoattractant protein-1 (MCP-1), were also significantly higher in KO compared with WT mice. These findings are the first to show that the PECAM-1-deficient mouse develops progressive nonalcoholic fatty liver disease (NAFLD), supporting a role for PECAM-1 as a negative regulator of NAFLD progression. Future examination of recently identified PECAM-1 allelic isoforms in humans as potential risk factors for developing NASH may be warranted.
Claudin-18: a dominant tight junction protein in Barrett's esophagus and likely contributor to its acid resistance
Authors: Jovov Biljana , Van Itallie Christina M. , Shaheen Nicholas J. , Carson Johnny L. , Gambling Todd M. , Anderson James M. , Orlando Roy C.
Published: October 2007

Barrett's esophagus (BE) is a specialized columnar epithelium (SCE) that develops as replacement for damaged squamous epithelium (SqE) in subjects with reflux disease, and as such it is apparently more acid resistant than SqE. How SCE resists acid injury is poorly understood; one means may involve altered tight junctions (TJs) since the TJ in SqE is an early target of attack and damage by acid in reflux disease. To assess this possibility, quantitative RT-PCR for 21 claudins was performed on endoscopic biopsies on SCE of BE and from healthy SqE from subjects without esophageal disease. In SCE, Cldn-18 was the most highly expressed at the mRNA level and this finding is paralleled by marked elevation in protein expression on immunoblots. In contrast in SqE, Cldn-18 was minimally expressed at the mRNA level and undetectable at the protein level. Immunofluorescence studies showed membrane localization of Cldn-18 and colocalization with the tight junction protein, zonula occludens-1. When Cldn-18 was overexpressed in MDCK II cells and mounted as monolayers in Ussing chambers, it raised electrical resistance and, as shown by lower dilution potentials to a NaCl gradient and lower diffusion potentials to acidic gradients, selectively reduced paracellular permeability to both Na+ and H+ compared with parental MDCK cells. We conclude that Cldn-18 is the dominant claudin in the TJ of SCE and propose that the change from a Cldn-18-deficient TJ in SqE to a Cldn-18-rich TJ in SCE contributes to the greater acid resistance of BE.
Cell cycle and apoptosis regulatory protein-1: a novel regulator of apoptosis in the colonic mucosa during aging
Authors: Majumdar Adhip P. N. , Du Jianhua , Yu Yingjie , Xu Hu , Levi Edi , Patel Bhaumik B. , Rishi Arun K.
Published: October 2007

Although the regulatory mechanisms for the age-related rise in proliferation and reduction in apoptosis in the colonic mucosa are yet to be fully delineated, we have demonstrated that these events are associated with increased expression and activation of epithelial growth factor receptor (EGFR)/ErbB-1 and some of its receptor family members (EGFRs), indicating their involvement in these processes. However, the downstream signaling events of EGFR and/or its family members regulating age-related changes in mucosal proliferation and apoptosis remain to be delineated. Cell cycle and apoptosis regulatory protein-1 (CARP-1), a novel growth signaling regulator that we isolated, participates in EGFR-dependent signaling. In the current investigation, we examined the involvement of CARP-1 in colonic mucosal growth-related processes during aging. We report that the age-related reduction in apoptosis in the colonic mucosa is associated with increased expression and tyrosine phosphorylation of not only EGFR but also ErbB-2 and ErbB-3. In contrast, protein and mRNA levels of CARP-1 as well as tyrosine phosphorylation of CARP-1 are decreased. Additionally, we have observed that administration of wortmannin, an inhibitor of phosphatidylinositol 3-kinase activity that accelerates apoptosis in the colonic mucosa of aged rats, causes a marked increase in expression and tyrosine phosphorylation of CARP-1. The age-related decline in CARP-1 expression could partly be attributed to increased methylation of the CARP-1 promoter. Taken together, our data suggest that not only EGFR but also its other members are involved in regulating colonic mucosal growth during aging and that CARP-1 may play a crucial role in transducing EGFRs signals.
Association of PepT1 with lipid rafts differently modulates its transport activity in polarized and nonpolarized cells
Authors: Nguyen Hang Thi Thu , Charrier-Hisamuddin Laetitia , Dalmasso Guillaume , Hiol Abel , Sitaraman Shanthi , Merlin Didier
Published: October 2007

The transporter PepT1, apically expressed in intestinal epithelial cells, is responsible for the uptake of di/tripeptides. PepT1 is also expressed in nonpolarized immune cells. Here we investigated the localization of PepT1 in lipid rafts in small intestinal brush border membranes (BBMs) and polarized and nonpolarized cells, as well as functional consequences of the association of PepT1 with lipid rafts. Immunoblot analysis showed the presence of PepT1 in low-density fractions isolated from mouse intestinal BBMs, polarized intestinal Caco2-BBE cells, and nonpolarized Jurkat cells by solubilization in ice-cold 0.5% Triton X-100 and sucrose gradient fractionation. PepT1 colocalized with lipid raft markers GM1 and N-aminopeptidase in intestinal BBMs and Caco2-BBE cell membranes. Disruption of lipid rafts with methyl-β-cyclodextrin (MβCD) shifted PepT1 from low- to high-density fractions. Remarkably, we found that MβCD treatment increased PepT1 transport activity in polarized intestinal epithelia but decreased that in intestinal BBM vesicles and nonpolarized immune cells. Mutational analysis showed that phenylalanine 293, phenylalanine 297, and threonine 281 in transmembrane segment 7 of the human di/tripeptide transporter, hPepT1, are important for the targeting to lipid rafts and transport activity of hPepT1. In conclusion, the association of PepT1 with lipid rafts differently modulates its transport activity in polarized and nonpolarized cells.
Are interstitial cells of Cajal plurifunction cells in the gut?
Authors: Sarna Sushil K.
Published: October 2007

The proposed functions of the interstitial cells of Cajal (ICC) are to 1) pace the slow waves and regulate their propagation, 2) mediate enteric neuronal signals to smooth muscle cells, and 3) act as mechanosensors. In addition, impairments of ICC have been implicated in diverse motility disorders. This review critically examines the available evidence for these roles and offers alternate explanations. This review suggests the following: 1) The ICC may not pace the slow waves or help in their propagation. Instead, they may help in maintaining the gradient of resting membrane potential (RMP) through the thickness of the circular muscle layer, which stabilizes the slow waves and enhances their propagation. The impairment of ICC destabilizes the slow waves, resulting in attenuation of their amplitude and impaired propagation. 2) The one-way communication between the enteric neuronal varicosities and the smooth muscle cells occurs by volume transmission, rather than by wired transmission via the ICC. 3) There are fundamental limitations for the ICC to act as mechanosensors. 4) The ICC impair in numerous motility disorders. However, a cause-and-effect relationship between ICC impairment and motility dysfunction is not established. The ICC impair readily and transform to other cell types in response to alterations in their microenvironment, which have limited effects on motility function. Concurrent investigations of the alterations in slow-wave characteristics, excitation-contraction and excitation-inhibition couplings in smooth muscle cells, neurotransmitter synthesis and release in enteric neurons, and the impairment of the ICC are required to understand the etiologies of clinical motility disorders.
SMIT2 mediates all myo-inositol uptake in apical membranes of rat small intestine
Authors: Aouameur Rym , Da Cal Sandra , Bissonnette Pierre , Coady Michael J. , Lapointe Jean-Yves
Published: October 2007

This study presents the characterization of myo-inositol (MI) uptake in rat intestine as evaluated by use of purified membrane preparations. Three secondary active MI cotransporters have been identified; two are Na+coupled (SMIT1 and SMIT2) and one is H+coupled (HMIT). Through inhibition studies using selective substrates such as d-chiro-inositol (DCI, specific for SMIT2) and l-fucose (specific for SMIT1), we show that SMIT2 is exclusively responsible for apical MI transport in rat intestine; rabbit intestine appears to lack apical transport of MI. Other sugar transport systems known to be present in apical membranes, such as SGLT1 or GLUT5, lacked any significant contribution to MI uptake. Functional analysis of rat SMIT2 activity, via electrophysiological studies in Xenopus oocytes, demonstrated similarities to the activities of SMIT2 from other species (rabbit and human) displaying high affinities for MI (0.150 ± 0.040 mM), DCI (0.31 ± 0.06 mM), and phlorizin (Pz; 0.016 ± 0.007 mM); low affinity for glucose (36 ± 7 mM); and no affinity for l-fucose. Although these functional characteristics essentially confirmed those found in rat intestinal apical membranes, a unique discrepancy was seen between the two systems studied in that the affinity constant for glucose was ∼40-fold lower in vesicles ( Ki= 0.94 ± 0.35 mM) than in oocytes. Finally, the transport system responsible for the basolateral efflux transporter of glucose in intestine, GLUT2, did not mediate any significant radiolabeled MI uptake in oocytes, indicating that this transport system does not participate in the basolateral exit of MI from small intestine.
The role of caveolin-1 in PCB77-induced eNOS phosphorylation in human-derived endothelial cells
Authors: Lim Eun Jin , Smart Eric J. , Toborek Michal , Hennig Bernhard
Published: October 2007

Polychlorinated biphenyls (PCBs) may contribute to the pathology of atherosclerosis by activating inflammatory responses in vascular endothelial cells. Endothelial nitric oxide synthase (eNOS) is colocalized with caveolae and is a critical regulator of vascular homeostasis. PCBs may be proatherogenic by causing dysfunctional eNOS signaling. The objective of this study was to investigate the role of caveolin-1 in PCB-induced endothelial dysfunction with a focus on mechanisms associated with eNOS signaling. Cells derived from an immortalized human vascular endothelial cell line were treated with PCB77 to study nitrotyrosine formation through eNOS signaling. Phosphorylation studies of eNOS, caveolin-1, and kinases, such as Src, phosphatidylinositol 3-kinase (PI3K), and Akt, were conducted in cells containing either functional or small-interfering RNA-silenced caveolin-1 protein. We also investigated caveolin-1-regulated mechanisms associated with PCB-induced markers of peroxynitrite formation and DNA binding of NF-κB. Cellular exposure to PCB77 increased eNOS phosphorylation and nitric oxide production, as well as peroxynitrite levels. A subsequent PCB-induced increase in NF-κB DNA binding may have implications in oxidative stress-mediated inflammatory mechanisms. The activation of eNOS by PCB77 treatment was blocked by inhibitors of the Src/PI3K/Akt pathway. PCB77 also increased phosphorylation of caveolin-1, indicating caveolae-dependent endocytosis. Caveolin-1 silencing abolished both the PCB-stimulated Akt and eNOS phosphorylation, suggesting a regulatory role of caveolae in PCB-induced eNOS signaling. These findings suggest that PCB77 induces eNOS phosphorylation in endothelial cells through a Src/PI3K/Akt-dependent mechanism, events regulated by functional caveolin-1. Our data provide evidence that caveolae may play a critical role in regulating vascular endothelial cell activation and toxicity induced by persistent environmental pollutants such as coplanar PCBs.
Forearm blood flow follows work rate during submaximal dynamic forearm exercise independent of sex
Authors: Gonzales Joaquin U. , Thompson Benjamin C. , Thistlethwaite John R. , Harper Allison J. , Scheuermann Barry W.
Published: October 2007

To test the hypothesis that sex influences forearm blood flow (FBF) during exercise, 15 women and 16 men of similar age [women 24.3 ± 4.0 (SD) vs. men 24.9 ± 4.5 yr] but different forearm muscle strength (women 290.7 ± 44.4 vs. men 509.6 ± 97.8 N; P < 0.05) performed dynamic handgrip exercise as the same absolute workload was increased in a ramp function (0.25 W/min). Task failure was defined as the inability to maintain contraction rate. Blood pressure and FBF were measured on separate arms during exercise by auscultation and Doppler ultrasound, respectively. Muscle strength was positively correlated with endurance time ( r = 0.72, P < 0.01) such that women had a shorter time to task failure than men (450.5 ± 113.0 vs. 831.3 ± 272.9 s; P < 0.05). However, the percentage of maximal handgrip strength achieved at task failure was similar between sexes (14% maximum voluntary contraction). FBF was similar between women and men throughout exercise and at task failure (women 13.6 ± 5.3 vs. men 14.5 ± 4.9 ml·min−1·100 ml−1). Mean arterial pressure was lower in women at rest and during exercise; thus calculated forearm vascular conductance (FVC) was higher in women during exercise but similar between sexes at task failure (women 0.13 ± 0.05 vs. men 0.11 ± 0.04 ml·min−1·100 ml−1·mmHg−1). In conclusion, the similar FBF during exercise was achieved by a higher FVC in the presence of a lower MAP in women than men. Still, FBF remained coupled to work rate (and presumably metabolic demand) during exercise irrespective of sex.
Amino acid substitutions at proline 220 of β-tubulin confer resistance to paclitaxel and colcemid
Authors: Yin Shanghua , Cabral Fernando , Veeraraghavan Sudha
Published: October 2007

Chinese hamster ovary cells selected for resistance to paclitaxel have a high incidence of mutations affecting L215, L217, and L228 in the H6/H7 loop region of β1-tubulin. To determine whether other mutations in this loop are also capable of conferring resistance to drugs that affect microtubule assembly, saturation mutagenesis of the highly conserved P220 codon in β1-tubulin cDNA was carried out. Transfection of a mixed pool of plasmids encoding all possible amino acid substitutions at P220 followed by selection in paclitaxel produced cell lines containing P220L and P220V substitutions. Similar selections in colcemid, on the other hand, yielded cell lines with P220C, P220S, and P220T substitutions. Site-directed mutagenesis and retransfection confirmed that these mutations were responsible for drug resistance. Expression of tubulin containing the P220L and P220V mutations reduced microtubule assembly, conferred resistance to paclitaxel and epothilone A, but increased sensitivity to colcemid and vinblastine. In contrast, tubulin with the P220C, P220S, and P220T mutations increased microtubule assembly, conferred resistance to colcemid and vinblastine, but increased sensitivity to paclitaxel and epothilone A. The results are consistent with molecular modeling studies and support a drug resistance mechanism based on changes in microtubule assembly that counteract the effects of drug treatment. These studies show for the first time that different substitutions at the same amino acid residue in β1-tubulin can confer cellular resistance to either microtubule-stabilizing or microtubule-destabilizing drugs. [Mol Cancer Ther 2007;6(10):2798–806]
Treatment of metastatic renal cancer with capsid-modified oncolytic adenoviruses
Authors: Guse Kilian , Ranki Tuuli , Ala-Opas Martti , Bono Petri , Särkioja Merja , Rajecki Maria , Kanerva Anna , Hakkarainen Tanja , Hemminki Akseli
Published: October 2007

Renal cancer is a common and deadly disease that lacks curative treatments when metastatic. Here, we have used oncolytic adenoviruses, a promising developmental approach whose safety has recently been validated in clinical trials. Although preliminary clinical efficacy data exist for selected tumor types, potency has generally been less than impressive. One important reason may be that expression of the primary receptor, coxsackie-adenovirus receptor, is often low on many or most advanced tumors, although not evaluated in detail with renal cancer. Here, we tested if fluorescence-assisted cell sorting could be used to predict efficacy of a panel of infectivity-enhanced capsid-modified marker gene expressing adenoviruses in renal cancer cell lines, clinical specimens, and subcutaneous and orthotopic murine models of peritoneally metastatic renal cell cancer. The respective selectively oncolytic adenoviruses were tested for killing of tumor cells in these models, and biodistribution after locoregional delivery was evaluated. In vivo replication was analyzed with noninvasive imaging. Ad5/3-Δ24, Ad5-Δ24RGD, and Ad5.pK7-Δ24 significantly increased survival of mice compared with mock or wild-type virus and 50% of Ad5/3-Δ24 treated mice were alive at 320 days. Because renal tumors are often highly vascularized, we investigated if results could be further improved by adding bevacizumab, a humanized antivascular endothelial growth factor antibody. The combination was well tolerated but did not improve survival, suggesting that the agents may be best used in sequence instead of together. These results set the stage for clinical testing of oncolytic adenoviruses for treatment of metastatic renal cancer currently lacking other treatment options. [Mol Cancer Ther 2007;6(10):2728–36]
Malignant mesothelioma cells are rapidly sensitized to TRAIL-induced apoptosis by low-dose anisomycin via Bim
Authors: Abayasiriwardana Keith S. , Barbone Dario , Kim Ki-Up , Vivo Claire , Lee Kevin K. , Dansen Tobias B. , Hunt Abigail E. , Evan Gerard I. , Broaddus V. Courtney
Published: October 2007

Tumor necrosis factor–related apoptosis inducing ligand (TRAIL) holds promise for the treatment of tumors; however, many tumors are resistant to TRAIL alone. We previously showed that resistant malignant mesothelioma cells are sensitized to TRAIL-induced apoptosis by diverse toxic insults including chemotherapy, irradiation, or protein translation inhibitors such as cycloheximide. In seeking nontoxic sensitizers for TRAIL, we tested the protein translation inhibitor anisomycin at subtoxic concentrations 10- to 100-fold below those reported to inhibit protein translation. At these low concentrations (25 ng/mL), anisomycin potently and rapidly sensitized mesothelioma cells to TRAIL-induced apoptosis. Moreover, such sensitization occurred in malignant but not in nonmalignant mesothelial cells. Sensitization by anisomycin was dependent on Bid, indicating a role for mitochondrial amplification in the apoptotic synergy with TRAIL signaling. Consistent with this, we found that anisomycin induces rapid accumulation of the BH3-only protein Bim; moreover, small interfering RNA knockdown of Bim inhibits anisomycin-induced sensitization. Bim accumulation seems not to be transcriptional; instead, it is associated with Bim phosphorylation and increased stability, both consistent with the activation of c-jun NH2-terminal kinase signals by anisomycin. Overall, our data indicate that the rapid and selective sensitization by anisomycin in mesothelioma cells is mediated by posttranslational potentiation of Bim, which primes the cells for apoptosis via the death receptor pathway. Such subtoxic approaches to sensitization may enhance the value of TRAIL in cancer therapy. [Mol Cancer Ther 2007;6(10):2766–76]
Soluble HLA-G and Embryo Implantation: Frequently Asked Questions
Authors: Le Bouteiller Philippe , Tabiasco Julie , Parinaud Jean
Published: October 2007

Two major criteria are currently used for optimal human embryo selection in in vitro fertilization or intracytoplasmic sperm injection: overall morphology-based grading system and rate of fragmentation. The measurement of soluble HLA-G (sHLA-G) in human preimplantation embryo culture supernatants gives great hope that addition of such a biochemical quantitative criterion would further improve the rates of implantation. In this short review, we will examine the functional significance of this latter approach by discussing the following frequently asked questions: (1) Which sHLA-G isoform(s) might be present in the human preimplantation embryo culture supernatants? (2) Where do these sHLA-G molecules come from? (3) Why are some preimplantation embryos ‘HLA-G secretors’ and others not? (4) How might the functions exerted by sHLA-G molecules influence embryonic implantation?
Comparative Implantation and Placentation
Authors: Chavatte-Palmer Pascale , Guillomot Michel
Published: October 2007

In all placental mammals, the establishment of an intimate contact between the embryo and the mother follows a succession of common critical steps whose chronology and timing may considerably vary from species to species. These processes present a great diversity based on the anatomy and the histology of the uterus, the developmental stage of the embryo at the time of implantation, and the endocrine and molecular interactions between the uterine and the embryonic tissues. This paper provides an overview of the general mechanical, endocrinological and cellular aspects involved in implantation in mammals with an emphasis on domestic species.
A Long-Term Follow-Up Study of Chest Pain Patients: Effect of Panic Disorder on Mortality, Morbidity, and Quality of Life
Authors: Bull Bringager Christine , Arnesen Harald , Friis Svein , Husebye Trygve , Dammen Toril
Published: October 2007

Aims: The aim was to assess the association between panic disorder (PD) and the long-term outcome of chest pain patients with or without coronary artery disease (CAD). Methods: Patients (n = 199) consecutively referred to a cardiology outpatient clinic because of chest pain were reassessed after 9 years. At the initial examination 16% suffered from CAD and 38% from PD. Data were collected on mortality, cardiac events, cardiac risk factors, chest pain, anxiety and depression (SCL-90-R), and health-related quality of life (SF-36). Results: The death rate in the study population was not significantly different from that in the general population and no significant associations were found between PD at baseline and mortality and cardiac morbidity at follow-up. PD was associated with significantly higher follow-up scores of chest pain intensity (p = 0.025), depression (p = 0.005), anxiety (p = 0.039), and poorer health-related quality of life: physical functioning (p = 0.004), role physical (p = 0.001), body pain (p = 0.007), and general health (p < 0.001). Conclusions: PD has a negative long-term effect on psychological and physical well-being of chest pain patients which emphasizes the necessity of identifying PD patients and offering them adequate treatment.
Identification of Atypical PML-RARA Breakpoint in a Patient with Acute Promyelocytic Leukemia
Authors: Ismail Said , Ababneh Nidaa , Awidi Abdalla
Published: October 2007

Acute promyelocytic leukemia (APL) of the M3 subtype is characterized by translocation t(15;17) that generates the PML-RARA fusion gene. Depending on the breakpoint position in the PML gene, 3 main fusion transcripts usually result. These breakpoints are bcr1 and bcr3 in introns 6 and 3, respectively, and bcr2 in exon 6. This report describes a rare atypical bcr2 breakpoint in a patient with morphological, cytogenetic and molecular features of APL. The presence of t(15;17) was first revealed by fluorescent in situ hybridization. Molecular analysis by reverse transcription polymerase chain reaction using primers for different PML-RARA junctions showed bands with different sizes compared with those generated from the three classical breakpoints, namely bcr1, bcr2 and bcr3. However, sequence analysis confirmed the presence of a bcr2 transcript with an atypical breakpoint within exon 6. The patient responded well to treatment and is now in complete remission. However, suggesting a favorable prognosis associated with such a rare transcript is difficult as more similar cases are needed to confirm such a conclusion. This article also stresses the importance of sequencing unusual polymerase chain reaction products to confirm their nature.
Primary Anaplastic Large Cell Lymphoma of the Lung
Authors: Yang Hong-Bo , Li Jian , Shen Ti
Published: October 2007

To our knowledge, only eleven cases of primary anaplastic large cell lymphoma (ALCL) of the lung have previously been reported. We describe here another two cases of primary pulmonary ALCL that developed in two Chinese women. Both patients presented with insidious symptoms related to pulmonary mass or nodule lesions. A comprehensive workup failed to show disease outside the chest. CD30-positive ALCL was demonstrated by histopathological studies of the lung tissue. Both patients were treated with 6 cycles of CHOP (cyclophosphamide, doxorubicin, vincristine and prednisone) chemotherapy and had complete remission. Primary pulmonary lymphoma is a great challenge for pneumologists since the clinical presentations and radiological findings are nonspecific. Appropriate invasive biopsy is necessary for early diagnosis.
Leopold Auenbrugger: Camphor-Induced Epilepsy – Remedy for Manic Psychosis
Authors: Pearce J.M.S.
Published: October 2007

Leopold Auenbrugger invented the art of diagnostic percussion. This paper recalls his use of camphor to induce epileptic fits, which were considered a remedy for psychosis. The initiation of the art of percussion by Auenbrugger is outlined, with a brief biographical sketch.
A Delayed Recrudescent Case of Sigmoidocutaneous Fistula due to Diverticulitis
Authors: Fujii Takaaki , Nakabayashi Toshihiro , Hashimoto Shinji , Kuwano Hiroyuki
Published: October 2007

Colocutaneous fistula caused by diverticulitis is relatively rare, and a delayed recrudescent case of colocutaneous fistula is very uncommon. We herein report a rare case of a Japanese 56-year-old male with delayed recrudescent sigmoidocutaneous fistula due to diverticulitis. A colocutaneous fistula was formed after a drainage operation against a perforation of the sigmoid colon diverticulum. After 5 years from treatment, he was admitted to our hospital because of lower abdominal pain. We diagnosed the recrudescent sigmoidocutaneous fistula by abdominal computed tomography and gastrografin enema, and managed the patient with total parenteral nutrition and antibiotics. As the fistula formation did not improve, a low anterior resection with fistulectomy was performed. The postoperative course was uneventful and the patient was discharged. It has been reported that, in fistulas of the skin caused by diverticular disease, complete closure of the fistula by conservative therapy may not be possible. This case also implies the possibility of a recurrence of the fistula even if the conservative treatment was effective. In cases of colocutaneous fistulas due to diverticulitis, radical surgery is considered necessary because of possibility of recurrence of the fistula.
An Entropy-Based Measure for QTL Mapping Using Extreme Samples of Population
Authors: Li Yu-Mei , Xiang Yang , Sun Zhen-Qiu
Published: October 2007

Quantitative trait locus (QTL) mapping can be accomplished through the method of selective genotyping, which is based on the differences of frequencies between an upper sample and a lower sample in population. However, amplifying the differences in marker allele frequencies in extreme samples may increase the probability for QTL mapping. Shannon entropy, which is a nonlinear function of allele frequencies, can be used to amplify the differences in marker allele frequencies. In this paper, we present a novel measure for linkage disequilibrium (LD) between a marker and single QTL, that is based on the comparison of the entropy and conditional entropy in a marker in extreme samples of population. This measure of LD between the marker and the trait locus can be used when the marker allele frequencies are known in the extreme samples of a population. We investigate the mapping performance in both analytic and simulation scenarios of a single QTL linked to a single marker. Our results show that the measure has very reasonable performance. In addition, a simulation study is performed on the basis of the haplotype frequencies of 10 SNPs of angiotensin-I converting enzyme (ACE) genes.
Vascular Mild Cognitive Impairment Is Highly Prevalent after Lacunar Stroke but Does Not Increase over Time: A 2-Year Follow-Up Study
Authors: Rasquin S.M.C. , van Oostenbrugge R.J. , Verhey F.R.J. , Lodder J.
Published: October 2007

Although ample research has been done into cognitive disorders occurring after stroke, relatively few data are available on the development and the course of vascularmild cognitive impairment (VMCI) after first-ever lacunar stroke. Methods: A cohort of 95 patients with a first-ever symptomaticlacunar infarct, older than 40 years, MMSE ≧15 and no other neurological or major psychiatric deficits were included. Patients were assessed (clinically and with a neuropsychological test battery) at 1 and 24 months after stroke, and CT was repeated. VMCI was diagnosed when patients had a deficit in at least one cognitive domain, in the absence of dementia. Results: Approximately 75% of the patients had VMCI at 1 month; this percentage was somewhat lower at 2 years. Only initial stroke severity was an independent predictor of VMCI after stroke. Conclusion: VMCI is highly prevalent after lacunar stroke, but does not increase during the first 2 years thereafter.
Antimalarial Drug Toxicity: A Review
Authors: AlKadi Hussien O.
Published: October 2007

Antimalarial drug toxicity is viewed differently depending upon whether the clinical indication is for malaria treatment or prophylaxis. In the treatment of Plasmodium falciparum malaria, which has a high mortality if untreated, a greater risk of adverse reactions to antimalarial drugs is inevitable. As chloroquine resistance has become widespread, alternative agents may be used in treatment regimens, however, the toxicity of these antimalarial agents should be considered. Quinine is the mainstay for treating severe malaria due to its rare cardiovascular or CNS toxicity, but its hypoglycemic effect may be problematic. Mefloquine can cause dose-related serious neuropsychiatric toxicity and pyrimethamine-dapsone is associated with agranulocytosis, especially if the recommended dose is exceeded. Pyrimethamine-sulfadoxine and amodiaquine are associated with a relatively high incidence of potentially fatal reactions, and are no longer recommended for prophylaxis. Atovaquone/proguanil is an antimalarial combination with good efficacy and tolerability as prophylaxis and for treatment. The artemisinin derivatives have remarkable efficacy and an excellent safety record. Prescribing in pregnancy is a particular problem for clinicians because the risk-benefit ratio is often very unclear.
Preferential Selection of IMP and VIM Metallo-β-Lactamases by Imipenem in Pseudomonas aeruginosa
Authors: Walsh Fiona , Bracher Simon , Turner Phil , Amyes Sebastian
Published: October 2007

Mixed populations of Pseudomonas aeruginosa, comprising a carbapenem-susceptible and a carbapenem-resistant strain, containing either the IMP or VIM β-lactamases, were exposed to imipenem. They developed resistance to both imipenem and meropenem. When the same mixed populations were exposed to meropenem, only the mixture containing the VIM β-lactamases became resistant to both carbapenems.
Corticotropin-Releasing Hormone and Pro-Opiomelanocortin Gene Expression in Female Monkeys with Differences in Sensitivity to Stress
Authors: Centeno Maria Luisa , Sanchez Rachel L. , Reddy Arubala P. , Cameron Judy L. , Bethea Cynthia L.
Published: October 2007

Background/Aims: The expressions of corticotropin-releasing hormone (CRH) and pro-opiomelanocortin (POMC) were assessed in brain tissue collected from nonstressed female cynomolgus monkeys previously categorized as highly stress resilient (HSR), medium stress resilient (MSR), or stress sensitive (SS) with respect to stress-induced anovulation. Methods: In situ hybridization and quantitative image analysis was used to measure mRNAs coding for CRH in the hypothalamic paraventricular nucleus (PVN) and thalamic center median-subfascicular complex (CM-Sf). Then, CRH neurons in the PVN were immunostained and the area of immunostaining was measured. Also, CRH fibers were immunostained in the central nucleus of the amygdala and the area of immunostaining was obtained. Finally, POMC mRNA expression was characterized in the hypothalamic infundibular nucleus. The groups were compared with ANOVA and Student-Newman-Keul’s (SNK) post hoc comparison. Results: CRH mRNA was significantly elevated in the caudal PVN in the MSR and SS animals compared to HSR animals (p < 0.05, SNK). There was a significant increase in average and total CRH-positive area in the MSR and SS groups compared to the HSR group (p < 0.05, SNK). There was also a significant increase in CRH volume in the MSR and SS groups compared to the HSR group (p < 0.05, SNK). In the CM-Sf, the average CRH optical density was significantly higher in the MSR and SS groups than in the HSR group (p < 0.05, SNK). In the central nucleus of the amygdala, the area of CRH fiber staining was significantly higher in the SS group than in the MSR or HSR groups (p < 0.05, SNK). There was no difference between the groups in POMC mRNA expression in the mediobasal hypothalamus. Conclusion: Macaques that exhibit immediate suppression of reproductive function upon stress are considered stress sensitive. These animals have elevated CRH in the hypothalamus and limbic structures, which may play a role in suppressing the hypothalamic-gonadal axis upon stress initiation.
A Novel Class of Prolyl Hydroxylase Inhibitors Induces Angiogenesis and Exerts Organ Protection Against Ischemia
Authors: Nangaku Masaomi , Izuhara Yuko , Takizawa Shunya , Yamashita Toshiharu , Fujii-Kuriyama Yoshiaki , Ohneda Osamu , Yamamoto Masayuki , van Ypersele de Strihou Charles , Hirayama Noriaki , Miyata Toshio
Published: October 2007

Objective—

Hypoxia inducible factor (HIF) plays a pivotal role in the adaptation to ischemic conditions. Its activity is modulated by an oxygen-dependent hydroxylation of proline residues by prolyl hydroxylases (PHD).

Methods and Results—

We discovered 2 unique compounds (TM6008 and TM6089), which inhibited PHD and stabilized HIF activity in vitro. Our docking simulation studies based on the 3-dimensional structure of human PHD2 disclosed that they preferentially bind to the active site of PHD. Whereas PHD inhibitors previously reported inhibit PHD activity via iron chelation, TM6089 does not share an iron chelating motif and is devoid of iron chelating activity. In vitro Matrigel assays and in vivo sponge assays demonstrated enhancement of angiogenesis by local administration of TM6008 and TM6089. Their oral administration stimulated HIF activity in various organs of transgenic rats expressing a hypoxia-responsive reporter vector. No acute toxicity was observed up to 2 weeks after a single oral dose of 2000 mg/kg for TM6008. Oral administration of TM6008 protected neurons in a model of cerebrovascular disease. The protection was associated with amelioration of apoptosis but independent of enhanced angiogenesis.

Conclusions—

The present study uncovered beneficial effects of novel PHD inhibitors preferentially binding to the active site of PHD.
Macular Pigment Lutein Is Antiinflammatory in Preventing Choroidal Neovascularization
Authors: Izumi-Nagai Kanako , Nagai Norihiro , Ohgami Kazuhiro , Satofuka Shingo , Ozawa Yoko , Tsubota Kazuo , Umezawa Kazuo , Ohno Shigeaki , Oike Yuichi , Ishida Susumu
Published: October 2007

Background—

Choroidal neovascularization (CNV) is a critical pathogenesis in age-related macular degeneration, the most common cause of blindness in the developed countries. The aim of the current study was to investigate the effect of lutein supplementation on the development of the murine model of laser-induced CNV together with underlying molecular mechanisms.

Methods and Results—

Mice were orally pretreated with lutein daily from 3 days before laser photocoagulation untill the end of the study. The index of CNV volume was significantly suppressed by the treatment with lutein, compared with vehicle-treated animals. Lutein treatment led to significant inhibition of macrophage infiltration into CNV and of the in vivo and in vitro expression of inflammation-related molecules including vascular endothelial growth factor, monocyte chemotactic protein −1, and intercellular adhesion molecule-1. Importantly, lutein suppressed IκB-α degradation and nuclear translocation of nuclear factor (NF)-κB p65 both in vivo and in vitro. Additionally, the development of CNV was significantly suppressed by inhibiting NF-κB p65 nuclear translocation, to the levels seen in the lutein treatment.

Conclusions—

Lutein treatment led to significant suppression of CNV development together with inflammatory processes including NF-κB activation and subsequent upregulation of inflammatory molecules, providing molecular evidence of potential validity of lutein supplementation as a therapeutic strategy to suppress CNV.
Gamma-Glutamyltransferase Is Associated With Incident Vascular Events Independently of Alcohol Intake
Authors: Fraser Abigail , Harris Ross , Sattar Naveed , Ebrahim Shah , Smith George Davey , Lawlor D.A.
Published: October 2007

Objective—

To investigate the association of γ-glutamyltransferase (GGT) with incident CHD and stroke. GGT is a marker of alcohol intake but may also reflect oxidative stress and nonalcoholic fatty liver disease. Alanine aminotransferase (ALT) is the enzyme most closely associated with liver fat content.

Methods and Results—

Associations of GGT and ALT with incident CHD, stroke, and a combined outcome of CHD or stroke were examined in the British Women’s Heart and Health study (n=2961), and a meta-analysis of population based studies examining these associations was performed. In pooled analyses of fully adjusted results of 10 prospective studies, a change of 1 U/L of GGT was associated with a HR=1.20 (95% CI: 1.02, 1.40) for CHD; a HR=1.54 (95% CI: 1.20, 2.00) for stroke; and HR=1.34 (95% CI: 1.22, 1.48) for CHD or stroke. Heterogeneity was substantially decreased when 2 studies in Asian populations were excluded. In a subgroup of nondrinkers results were similar to the main analysis. Meta analyses of the only 2 studies that examined the association of ALT with incident cardiovascular events found a HR=1.18 (95% CI: 0.99, 1.41) for CHD and a HR=1.10 (95% CI: 0.89, 1.36) for CHD or stroke (combined).

Conclusion—

GGT is associated with incident vascular events independently of alcohol intake. The mechanisms underlying this association remain unclear and require future study.
Adiponectin Mediates the Suppressive Effect of Rosiglitazone on Plasminogen Activator Inhibitor-1 Production
Authors: Hoo Ruby L.C. , Chow W.S. , Yau M.H. , Xu A. , Tso Annette W.K. , Tse H.F. , Fong Carol H.Y. , Tam Sidney , Chan Lawrence , Lam Karen S.L.
Published: October 2007

Objective—

The purpose of this study was to examine the effects of PPAR-γ agonist rosiglitazone, relative to sulfonylureas, on circulating levels of adiponectin and the prothrombotic factor, plasminogen activator inhibitor (PAI)-1, in type 2 diabetic patients, and to investigate, in animal models, whether the antithrombotic action of rosiglitazone was mediated through adiponectin.

Methods and Results—

Our clinical study (n=64) showed that after 24-week add-on therapy, the rosiglitazone group had a greater mean reduction in plasma PAI-1 levels (25%, versus 12% in sulfonylurea group,
P
=0.002). Stepwise multiple linear regression analysis identified the reduction in plasma fasting glucose and the rise in adiponectin levels to be independently associated with the reduction in PAI-I concentration in the rosiglitazone-treated patients. Rosiglitazone (20 mg/kg/d) reduced adipose tissue PAI-1 mRNA expression and its plasma levels in wild-type C57 mice with diet-induced obesity (
P
<0.001), but this suppressive effect was attenuated in adiponectin knockout mice. Adenovirus-mediated overexpression of adiponectin led to a significant suppression of adipose tissue PAI-1 expression and its circulating concentrations in db/db diabetic mice. Our in vitro study demonstrated that recombinant adiponectin directly inhibited PAI-1 production in 3T3-L1 adipocytes.

Conclusions—

The antithrombotic effect of rosiglitazone is mediated, at least in part, through the suppressive effect of adiponectin on PAI-1 production.
Heritability of Clot Formation, Morphology, and Lysis
Authors: Carter Angela M. , Cymbalista Charlotte M. , Spector Tim D. , Grant Peter J.
Published: October 2007

Objective—

The relative balance between clot formation and fibrinolysis is considered to reflect thrombotic potential following vascular injury. The aims of the present study were to (1) to determine the contribution of genetic and environmental factors to variance in measures of clot structure/function in the Leeds Family Study, and (2) to determine the relationship between measures of clot structure/function and cardiovascular risk.

Methods and Results—

Using high throughput turbidimetric assays, heritabilities of measures of clot formation, clot structure, and clot lysis were ≈0.30. Fibrinogen contributed to variance in all measures and plasminogen activator inhibitor-1 to variance in lysis variables. Subjects at increased cardiovascular risk due to the presence of the metabolic syndrome (MetS) had increased clot density (MaxAbs
C
: 0.358 [0.340, 0.375]au) and prolonged lysis times (Lys
T
: 510 [6569, 7939]s) compared with those without MetS (MaxAbs
C
: 0.319 [0.310, 0.328]au,
P
=0.003; Lys
T
: 7221 [4884, 5328]s,
P
<0.001). Furthermore, measures of clot structure/function increased progressively with increasing number of MetS components.

Conclusions—

This study indicates that genetic factors contribute modestly to variance in clot structure/function and that clot structure/function is related to presence of the MetS and number of MetS components. Identification of the genetic and environmental factors influencing clot structure/function may further our understanding of the underlying factors predisposing to cardiovascular disease.
Effects of Aging and Hypoxia-Inducible Factor-1 Activity on Angiogenic Cell Mobilization and Recovery of Perfusion After Limb Ischemia
Authors: Bosch-Marce Marta , Okuyama Hiroaki , Wesley Jacob B. , Sarkar Kakali , Kimura Hideo , Liu Ye V. , Zhang Huafeng , Strazza Marianne , Rey Sergio , Savino Lindsey , Zhou Yi Fu , McDonald Karin R. , Na Youn , Vandiver Scott , Rabi Alireza , Shaked Yuval , Kerbel Robert , LaVallee Theresa , Semenza Gregg L.
Published: October 2007

Ischemia is a stimulus for production of angiogenic cytokines that activate local vascular cells and mobilize angiogenic cells to the circulation. These responses are impaired in elderly patients with peripheral arterial disease. Hypoxia-inducible factor (HIF)-1 mediates adaptive responses to ischemia, including production of angiogenic cytokines. In this study, we demonstrate that aging and HIF-1 loss-of-function impair the expression of multiple angiogenic cytokines, mobilization of angiogenic cells, maintenance of tissue viability, and recovery of limb perfusion following femoral artery ligation. We show that HIF-1 directly activates transcription of the gene encoding stem cell factor and that mice lacking the cognate receptor C-KIT have impaired recovery from ischemia. Administration of AdCA5, an adenovirus encoding a constitutively active form of HIF-1α, improved the recovery of perfusion in older mice to levels similar to those in young mice. Injection of AdCA5 into nonischemic limb was sufficient to increase the number of circulating angiogenic cells. These results indicate that HIF-1 activity is necessary and sufficient for the mobilization of angiogenic cells and that HIF-1α gene therapy can counteract the pathological effects of aging in a mouse model of limb ischemia.