bluuebunny/crossref_metadata_2025 huggingface dataset:
Hepatotoxicity Despite Early Administration of Intravenous N‐Acetylcysteine for Acute Acetaminophen Overdose
Authors: Doyon Suzanne , Klein‐Schwartz Wendy
Published: December 2008

Objectives:  The objective was to evaluate the effectiveness of intravenous N‐acetylcysteine (IV NAC; 300 mg/kg over 21 hours) in early acute acetaminophen (APAP) overdose patients.

Methods:  This observational case series included patients hospitalized between 2004 and 2007 for acute APAP overdoses and who were reported to a regional poison center. Inclusion criteria were plasma APAP concentrations on or above the treatment line on the Rumack‐Matthew nomogram, administration of IV NAC within 8 hours of ingestion, and follow‐up to known outcome. The hospital chart of each patient who received IV NAC for longer than the standard 21 hours was reviewed. Hepatotoxicity was defined as hepatic aminotransferase levels greater than 1,000 IU/L.

Results:  Seventy‐seven patients met inclusion criteria and received at least 21 hours of IV NAC for an acute APAP overdose. Seven patients received antidotal therapy for greater than 21 hours. These patients tended to have ingested combination preparations, have very high initial plasma APAP concentrations, and had persistently elevated plasma concentrations during their hospital stay. Hepatotoxicity occurred in 4 patients (5.2%, 95% confidence interval [CI] = 0.2% to 10.1%), including 1 death and 1 liver transplantation.

Conclusions:  Hepatotoxicity developed in 5.2% of cases, suggesting that the 21‐hour IV NAC regimen is suboptimal in some patients. In addition to high initial plasma APAP concentrations, APAP product formulation and persistently elevated plasma APAP concentrations were identified as factors possibly associated with developing hepatotoxicity. The authors propose a tailored approach to the discontinuation of IV NAC and point out the need for reevaluation of optimal doses and duration of therapy.
Proteolytic‐Based Method for the Identification of Human Growth Hormone*
Authors: Wisniewski Eric S. , Rees David K. , Chege Esther W.
Published: December 2008

Abstract:  Human growth hormone (HGH) is a relatively small protein consisting of 191 amino acids and has an average mass of 22,125 amu. The forensic analysis of proteins such as HGH must meet the analytical sufficiency requirements for the laboratory and consists of a binary approach. A suspected sample is analyzed as the whole protein for retention time and mass determination using high performance liquid chromatography equipped with a photodiode array and liquid chromatography mass spectrometry. Further fragmentation of the protein using a proteolytic enzyme adds another dimension to the specificity of the analysis. Porcine trypsin digests proteins in a very predictable manner and yields peptide fragments of the original protein that can be used as a means for fingerprinting the larger biomolecule. In silico, or theoretical, digestion of HGH by trypsin yields 21 peptides ranging in size from 1 to 23 amino acids in length. The larger fragments containing higher numbers of amino acids give more specificity to identifying a protein based on a fragment produced by the digestion of trypsin. Herein, the analysis of HGH using a proteolytic approach is presented that meets the Scientific Working Group for the Analysis of Seized Drugs (SWGDRUG) recommendations for the identification of unknown substances.
Comparative Analysis of the HV1 and HV2 Regions of Human Mitochondrial DNA by Denaturing High‐Performance Liquid Chromatography
Authors: Kristinsson Richard , Lewis Sarah E. , Danielson Phillip B.
Published: December 2008

Abstract:  Denaturing high‐performance liquid chromatography (DHPLC) was evaluated as a sequencing‐independent means of detecting the presence of sequence differences in pair‐wise mixtures of nonconcordant amplicons of human mitochondrial DNA (mtDNA). A total of 920 pair‐wise combinations of HV1 and HV2 mtDNA amplicons from 95 individuals were assayed by DHPLC for sequence concordance/nonconcordance. For the 72 combinations of amplicons from different individuals who shared identical DNA sequences, DHPLC assays consistently indicated sequence concordance between the samples. This was in 100% agreement with sequencing data. For the 849 combinations of amplicons which differed in sequence, DHPLC detected the presence of sequence nonconcordance in all but 13 assays to yield 98.5% concordance with sequencing. Thus, DHPLC can be used to detect a diversity of sequence differences (transitions, transversions, insertions, and deletions) in the mtDNA D‐loop. Accordingly, DHPLC may have utility as a presumptive indicator of mtDNA sequence concordance samples, as a screen for heteroplasmy/situational mixtures, and as a means for the physical fractionation of the individual contributors to an mtDNA mixture prior to sequencing.
URG11 promotes gastric cancer growth and invasion by activation of β‐catenin signalling pathway
Authors: Du Rui , Xia Lin , Sun Shiren , Lian Zhaorui , Zou Xue , Gao Juan , Xie Huahong , Fan Rui , Song Jiugang , Li Xiaohua , Liu Jie , Fan Daiming
Published: December 2008

Upregulated gene 11 (URG11), a new gene upregulated by Heptatitis B Virus X protein (HBx), was previously shown to activate β‐catenin and promote hepatocellular growth and tumourigenesis. Although the oncogenic role of URG11 in the development of hepatocellular carcinoma has been well documented, its relevance to other human malignancies and the underlying molecular mechanisms remain largely unknown. Here we reported a novel function of URG11 to promote gastric cancer growth and metastasis. URG11 was found to be highly expressed in gastric cancer tissues compared with adjacent nontumourous ones by immunohistochemical staining and western blot. Knockdown of URG11 expression by small interfering RNA (siRNA) effectively attenuated the proliferation, anchorage‐independent growth, invasiveness and metastatic potential of gastric cancer cells. URG11 inhibition led to decreased expression of β‐catenin and its nuclear accumulation in gastric cancer cells and extensive costaining between URG11 and β‐catenin was observed in gastric cancer tissues. Transient transfection assays with the β‐catenin promoter showed that it was inhibited by URG11‐specific small inhibitory RNA. Moreover, suppression of endogenous URG11 expression results in decreased activation of β‐catenin/TCF and its downstream effector genes, cyclinD1 and membrane type 1 matrix metallopeptidase (MT1‐MMP), which are known to be involved in cell proliferation and invasion, respectively. Taken together, our data suggest that URG11 contributes to gastric cancer growth and metastasis at least partially through activation of β‐catenin signalling pathway. These findings also propose a promising target for gene therapy in gastric cancer.
The α1HCa2+channel subunit is expressed in mouse jejunal interstitial cells of Cajal and myocytes
Authors: Gibbons Simon J. , Strege Peter R. , Lei Sha , Roeder Jaime L. , Mazzone Amelia , Ou Yijun , Rich Adam , Farrugia Gianrico
Published: December 2008

T‐type Ca2+currents have been detected in cells from the external muscular layers of gastrointestinal smooth muscles and appear to contribute to the generation of pacemaker potentials in interstitial cells of Cajal from those tissues. However, the Ca2+channel α subunit responsible for these currents has not been determined. We established that the α subunit of the α1HCa2+channel is expressed in single myocytes and interstitial cells of Cajal using reverse transcription and polymerase chain reaction from whole tissue, laser capture microdissected tissue and single cells isolated from the mouse jejunum. Whole‐cell voltage clamp recordings demonstrated that a nifedipine and Cd2+resistant, mibefradil‐sensitive current is present in myocytes dissociated from the jejunum. Electrical recordings from the circular muscle layer demonstrated that mibefradil reduced the frequency and initial rate of rise of the electrical slow wave. Gene targeted knockout of both alleles of thecacna1hgene, which encodes the α1HCa2+channel subunit, resulted in embryonic lethality because of death of the homozygous knockouts prior to E13.5 daysin utero. We conclude that a channel with the pharmacological and molecular characteristics of the α1HCa2+channel subunit is expressed in interstitial cells of Cajal and myocytes from the mouse jejunum, and that ionic conductances through the α1HCa2+channel contribute to the upstroke of the pacemaker potential. Furthermore, the survival of mice that do not express the α1HCa2+channel protein is dependent on the genetic background and targeting approach used to generate the knockout mice.
Minerval induces apoptosis in Jurkat and other cancer cells
Authors: Llado Victoria , Gutierrez Antonio , Martínez Jordi , Casas Jesús , Terés Silvia , Higuera Mónica , Galmés Antonio , Saus Carles , Besalduch Joan , Busquets Xavier , Escribá Pablo V.
Published: December 2008

Minerval is an oleic acid synthetic analogue that impairs lung cancer (A549) cell proliferation upon modulation of the plasma membrane lipid structure and subsequent regulation of protein kinase C localization and activity. However, this mechanism does not fully explain the regression of tumours induced by this drug in animal models of cancer. Here we show that Minerval also induced apoptosis in Jurkat T‐lymphoblastic leukaemia and other cancer cells. Minerval inhibited proliferation of Jurkat cells, concomitant with a decrease of cyclin D3 and cdk2 (cyclin‐dependent kinase2). In addition, the changes that induced on Jurkat cell membrane organization caused clustering (capping) of the death receptor Fas (CD95), caspase‐8 activation and initiation of the extrinsic apoptosis pathway, which finally resulted in programmed cell death. The present results suggest that the intrinsic pathway (associated with caspase‐9 function) was activated downstream by caspase‐8. In a xenograft model of human leukaemia, Minerval also inhibited tumour progression and induced tumour cell death. Studies carried out in a wide variety of cancer cell types demonstrated that apoptosis was the main molecular mechanism triggered by Minerval. This is the first report on the pro‐apoptotic activity of Minerval, and in part explains the effectiveness of this non‐toxic anticancer drug and its wide spectrum against different types of cancer.
In GERD patients, mucosal repair associated genes are upregulated in non‐inflamed oesophageal epithelium
Authors: De Vries D. R. , Ter Linde J. J. M. , Van Herwaarden M. A. , Schwartz M. P. , Shephard P. , Geng M. M. , Smout A. J. P. M. , Samsom M.
Published: December 2008

Previous studies addressing the effects of acid reflux and PPI therapy on gene expression in oesophageal epithelium concentrated on inflamed tissue. We aimed to determine changes in gene expression in non‐inflamed oesophageal epithelium of GERD patients. Therefore, we included 20 GERD patients with pathological total 24‐hr acid exposure of 6–12% and SAP ≥ 95%. Ten patients discontinued PPI treatment (PPI‐), 10 took pantoprazole 40 mg bid (PPI+). Ten age/sex‐matched healthy controls were recruited. Biopsies were taken from non‐inflamed mucosa 6 cm and 16 cm proximal to the squamocolumnar junction (SCJ). Gene expression profiling of biopsies from 6 cm was performed on Human Genome U133 Plus 2.0 arrays (Affymetrix). Genes exhibiting a fold change >1.4 (t‐test P‐value < 1E– 4) were considered differentially expressed. Results were confirmed by real‐time RT‐PCR. In PPI‐ patients, 92 microarray probesets were deregulated. The majority of the corresponding genes were associated with cell–cell contacts, cytoskeletal reorganization and cellular motility, suggesting facilitation of a migratory phenotype. Genes encoding proteins with anti‐apoptotic or anti‐proliferative functions or stress‐protective functions were also deregulated. No probesets were deregulated in PPI+ patients. QPCR analysis of 20 selected genes confirmed most of the deregulations in PPI‐ patients, and showed several deregulated genes in PPI+ patients as well. In the biopsies taken at 16 cm QPCR revealed no deregulations of the selected genes. We conclude that upon acid exposure, oesophageal epithelial cells activate a process globally known as epithelial restitution: up‐regulation of anti‐apoptotic, anti‐oxidant and migration associated genes. Possibly this process helps maintaining barrier function.
Pharmacological induction of vascular extracellular superoxide dismutase expression in vivo
Authors: Oppermann Marc , Balz Vera , Adams Volker , Thao‐Vi Dao Vu , Bas Murat , Suvorava Tatsiana , Kojda Georg
Published: December 2008

Pentaerythritol tetranitrate (PETN) treatment reduces progression of atherosclerosis and endothelial dysfunction and decreases oxidation of low‐density lipoprotein (LDL) in rabbits. These effects are associated with decreased vascular superoxide production, but the underlying molecular mechanisms remain unknown. Previous studies demonstrated that endogenous nitric oxide could regulate the expression of extracellular superoxide dismutase (ecSOD) in conductance vessels in vivo. We investigated the effect of PETN and overexpression of endothelial nitric oxide synthase (eNOS++) on the expression and activity of ecSOD. C57BL/6 mice were randomized to receive placebo or increasing doses of PETN for 4 weeks and eNOS++ mice with a several fold higher endothelial‐specific eNOS expression were generated. The expression of ecSOD was determined in the lung and aortic tissue by real‐time PCR and Western blot. The ecSOD activity was measured using inhibition of cytochrome C reduction. There was no effect of PETN treatment or eNOS overexpression on ecSOD mRNA in the lung tissue, whereas ecSOD protein expression increased from 2.5‐fold to 3.6‐fold (P < 0.05) by 6 mg PETN/kg body weight (BW)/day and 60 mg PETN/kg BW/day, respectively. A similar increase was found in aortic homogenates. eNOS++ lung cytosols showed an increase of ecSOD protein level of 142 ± 10.5% as compared with transgene‐negative littermates (P < 0.05), which was abolished by Nω‐nitro‐L‐arginine treatment. In each animal group, the increase of ecSOD expression was paralleled by an increase of ecSOD activity. Increased expression and activity of microvascular ecSOD are likely induced by increased bioavailability of vascular nitric oxide. Up‐regulation of vascular ecSOD may contribute to the reported antioxidative and anti‐atherosclerotic effects of PETN.
Enforced expression of protein kinase C in skeletal muscle causes physical inactivity, fatty liver and insulin resistance in the brain
Authors: Hennige Anita M. , Heni Martin , Machann Jürgen , Staiger Harald , Sartorius Tina , Hoene Miriam , Lehmann Rainer , Weigert Cora , Peter Andreas , Bornemann Antje , Kroeber Stefan , Pujol Anna , Franckhauser Sylvie , Bosch Fatima , Schick Fritz , Lammers Reiner , Häring Hans‐Ulrich
Published: December 2008

Among the multitude of dysregulated signalling mechanisms that comprise insulin resistance in divergent organs, the primary events in the development of type 2 diabetes are not well established. As protein kinase C (PKC) activation is consistently present in skeletal muscle of obese and insulin resistant subjects, we generated a transgenic mouse model that overexpresses constitutively active PKC‐β2 in skeletal muscle to test whether activation of PKC is sufficient to cause an aversive whole‐body phenotype. Upon this genetic modification, increased serine phosphorylation in Irs1 was observed and followed by impaired 3H‐deoxy‐glucose uptake and muscle glycogen content, and transgenic mice exhibited insulin and glucose intolerance as they age. Muscle histochemistry revealed an increase in lipid deposition (intramyocellular lipids), and transgenic mice displayed impaired expression of transcriptional regulators of genes involved in fatty acid oxidation (peroxisome proliferator‐activated receptor‐γ, PGC‐1β, acyl‐CoA oxidase) and lipolysis (hormone‐sensitive lipase). In this regard, muscle of transgenic mice exhibited a reduced capacity to oxidize palmitate and contained less mitochondria as determined by citrate synthase activity. Moreover, the phenotype included a profound decrease in the daily running distance, intra‐abdominal and hepatic fat accumulation and impaired insulin action in the brain. Together, our data suggest that activation of a classical PKC in skeletal muscle as present in the pre‐diabetic state is sufficient to cause disturbances in whole‐body glucose and lipid metabolism followed by profound alterations in oxidative capacity, ectopic fat deposition and physical activity.
Polymorphisms in p53 and the p53 pathway: roles in cancer susceptibility and response to treatment
Authors: Hrstka Roman , Coates Philip J. , Vojtesek Borivoj
Published: December 2008

Introduction

SNPs inTP53exons altering amino‐acid sequence

Polymorphism inTP53introns

Linkage between the polymorphisms

Silent mutations and SNPs inTP53

Polymorphisms in other members of the p53 family

Polymorphisms in p53 target genes

Polymorphisms in p53 response elements

Concluding remarks

The p53 tumour suppressor protein lies at the crossroads of multiple cellular response pathways that control the fate of the cell in response to endogenous or exogenous stresses and inactivation of the p53 tumour suppressor signalling pathway is seen in most human cancers. Such aberrant p53 activity may be caused by mutations in theTP53gene sequence producing truncated or inactive mutant proteins, or by aberrant production of other proteins that regulate p53 activity, such as gene amplification and overexpression of MDM2 or viral proteins that inhibit or degrade p53. Recent studies have also suggested that inherited genetic polymorphisms in the p53 pathway influence tumour formation, progression and/or response to therapy. In some cases, these variants are clearly associated with clinico‐pathological variables or prognosis of cancer, whereas in other cases the evidence is less conclusive. Here, we review the evidence that common polymorphisms in various aspects of p53 biology have important consequences for overall tumour susceptibility, clinico‐pathology and prognosis. We also suggest reasons for some of the reported discrepancies in the effects of common polymorphisms on tumourigenesis, which relate to the complexity of effects on tumour formation in combination with other oncogenic changes and other polymorphisms. It is likely that future studies of combinations of polymorphisms in the p53 pathway will be useful for predicting tumour susceptibility in the human population and may serve as predictive biomarkers of tumour response to standard therapies.
Integrin α1β1 is involved in the differentiation into myofibroblasts in adult reactive tissues in vivo
Authors: Rodriguez Alejandro , Karen Jakob , Gardner Humphrey , Gerdin Bengt , Rubin Kristofer , Sundberg Christian
Published: December 2008

Connective tissue cell activation is of importance during reactive conditions such as solid tumour growth, wound healing and pannus formation in rheumatoid arthritis. Here, we have compared connective tissue cells of mesenchymal origin in human tissues from these conditions and their normal counterparts using a panel of cell‐type‐specific markers. In particular, we investigated variations of integrin expression among connective tissue cell phenotypes. Connective tissue cell populations were defined based on their association with the microvasculature and their expression of activation markers. The phenotype of these cells varied according to the type of pathological connective tissue examined. Our morphological data from human tissues suggested that the α1β1 integrin, a collagen/laminin receptor, is involved in the differentiation of precursor cells into myofibroblasts. To mechanistically investigate this hypothesis, we employed experimental models for carcinoma growth and wound healing utilizing α1 integrin‐deficient mice. The data confirmed that the α1β1 integrin is of importance not only for the differentiation of mesenchymal cells into myofibroblasts but also for the neovascularization and connective tissue organization and emphasize the importance of myofibroblasts in the pathophysiology of tissue repair, inflammation and tumour growth.
Circulating endothelial cells, microparticles and progenitors: key players towards the definition of vascular competence
Authors: Sabatier F. , Camoin‐Jau L. , Anfosso F. , Sampol J. , Dignat‐George F.
Published: December 2008

Introduction

Dynamics between endothelial injury and repair: the response to injury theory ‘revisited’

Emerging biomarkers reflecting the dynamics between endothelial injury and repair: from pathophysiology to clinical testing.

Endothelial lesion versus regeneration: towards the definition of ‘vascular competence’

Conclusion

The balance between lesion and regeneration of the endothelium is critical for the maintenance of vessel integrity. Exposure to cardiovascular risk factors (CRF) alters the regulatory functions of the endothelium that progresses from a quiescent state to activation, apoptosis and death. In the last 10 years, identification of circulating endothelial cells (CEC) and endothelial‐derived microparticles (EMP) in the circulation has raised considerable interest as non‐invasive markers of vascular dysfunction. Indeed, these endothelial‐derived biomarkers were associated with most of the CRFs, were indicative of a poor clinical outcome in atherothrombotic disorders and correlated with established parameters of endothelial dysfunction. CEC and EMP also behave as potential pathogenic vectors able to accelerate endothelial dysfunction and promote disease progression. The endothelial response to injury has been enlarged by the discovery of a powerful physiological repair process based on the recruitment of circulating endothelial progenitor cells (EPC) from the bone marrow. Recent studies indicate that reduction of EPC number and function by CRF plays a critical role in the progression of cardiovascular diseases. This EPC‐mediated repair to injury response can be integrated into a clinical endothelial phenotype defining the ‘vascular competence’ of each individual. In the future, provided that standardization of available methodologies could be achieved, multimarker strategies combining CEC, EMP and EPC levels as integrative markers of ‘vascular competence’ may offer new perspectives to assess vascular risk and to monitor treatment efficacy.
Membrane‐targeted strategies for modulating APP and Aβ‐mediated toxicity
Authors: Price Katherine A. , Crouch Peter J. , Donnelly Paul S. , Masters Colin L. , White Anthony R. , Curtain Cyril C.
Published: December 2008

Introduction

Role of amyloid‐β (Aβ) and APP in AD

Cell membranes

Aβ and cell membranes

AD therapeutics targeting the membrane

Potential for metal complex delivered metal ions to alter membrane‐mediated activation pathways

Conclusions

Alzheimer's disease (AD) is a progressive neurodegenerative disease characterized by numerous pathological features including the accumulation of neurotoxic amyloid‐β (Aβ) peptide. There is currently no effective therapy for AD, but the development of therapeutic strategies that target the cell membrane is gaining increased interest. The amyloid precursor protein (APP) from which Aβ is formed is a membrane‐bound protein, and Aβ production and toxicity are both membrane mediated events. This review describes the critical role of cell membranes in AD with particular emphasis on how the composition and structure of the membrane and its specialized regions may influence toxic or benign Aβ/APP pathways in AD. The putative role of copper (Cu) in AD is also discussed, and we highlight how targeting the cell membrane with Cu complexes has therapeutic potential in AD.
Laminin isoforms in human embryonic stem cells: synthesis, receptor usage and growth support
Authors: Vuoristo Sanna , Virtanen Ismo , Takkunen Minna , Palgi Jaan , Kikkawa Yamato , Rousselle Patricia , Sekiguchi Kiyotoshi , Tuuri Timo , Otonkoski Timo
Published: December 2008

To reveal the functional intrinsic niche of human embryonic stem cells (hESC) we examined the production of basement membrane (BM) proteins and the presence of their receptors in feeder‐free cell culture conditions. In addition, we investigated binding of hESCs to purified human BM proteins and identified the receptors mediating these contacts. Also, we tested whether purified human laminin (Lm) isoforms have a role in hESC self‐renewal and growth in short‐term cultures. The results show that hESCs synthesize Lm α1 and Lm α5 chains together with Lm β1 and γ1 chains suggesting the production of Lms‐111 and ‐511 into the culture medium and deposits on cells. hESCs contain functionally important integrin (Int) subunits, Int β1, α3, α6, α5, β5 and αV, as well as the Lm α5 receptor, Lutheran (Lu) glycoprotein and its truncated form, basal cell adhesion molecule (B‐CAM). In cell adhesion experiments, Int β1 was crucial for adhesion to most of the purified human BM proteins. Lu/B‐CAM mediated adhesion to Lm‐511 together with Int α3β1, and was essential for the adhesion of hESCs to embryonic feeder cells. Adhesion to Lm‐411 was mediated by Int α6β1. Lm‐511 supported hESC growth in defined medium equally well as Matrigel. These results provide consequential information of the biological role of BM in hESCs, warranting further investigation of BM biology of human pluripotent stem cells.
Pathological findings in rats with experimental allergic encephalomyelitis
Authors: DONG MEI , LIU RUICHUN , GUO LI , LI CHUNYAN , TAN GUOJUN
Published: December 2008

The aim of this study was to establish an animal model of experimental allergic encephalomyelitis (EAE) and examine the basic pathological changes, as well as expression and distribution of MMP‐2 and MMP‐9, in Wistar rats. Tissue sections were processed for HE staining, Weil myelin staining, and modified Bielschowsky staining. Expression and distribution of glial fibrillary acidic protein (GFAP), matrix metalloproteinase‐2 (MMP‐2) and matrix metalloproteinase‐9 (MMP‐9) were detected with immunohistochemistry. We divided the EAE into five types, depending on pathological characteristics and clinical manifestations: acute EAE, relapsing‐remitting EAE, progressive EAE, benign EAE, and asymptomatic EAE. Rats with acute EAE suffered from quick, severe attacks with widespread inflammatory cells and axonal loss. No demyelination or astrocytic hyperplasia was found around the lesions. Rats with relapsing‐remitting EAE broke down twice, with many perivascular cuffs and demyelinating plaques in lesions; hyperplastic and hypertrophic astrocytes characterized old lesions and axonal loss was evident. Rats suffering from progressive EAE exhibited continuous aggravation without improvement, accompanied by perivascular cuffs, demyelination, increased gliocytes and axonal damage. Rats with benign EAE recovered to a normal state with obviously decreased inflammatory cells and almost entirely unaffected myelin and axons. Rats with asymptomatic EAE also had various pathological changes that were not coincident with their clinical manifestations. Elevated expression of MMP‐2 and MMP‐9 was concordant in different types of EAE, but the extent differed in each type of EAE. MMP‐2 and MMP‐9 can be expressed in the form of vascular endothelial cells, meninges, or accumulated inflammatory cells. Multiple clinical courses of disease were demonstrated in Wistar rat EAE, with attributes similar to multiple sclerosis (MS) in clinical and pathological characteristics. Elevated expression of MMP‐2 and MMP‐9 may play a role in some aspects of pathological changes in EAE, for example, destroying the blood‐brain barrier, degrading the myelin sheath, and damaging axons.
Intracellular replication of fusobacteria requires new actin filament formation of epithelial cells
Authors: GURSOY ULVI KAHRAMAN , KÖNÖNEN EIJA , UITTO VELI‐JUKKA
Published: December 2008

We examined survival and replication of fusobacteria inside epithelial cells. Subconfluent cultures of HaCaT keratinocytes were infected with five bacterial strains representing three Fusobacterium species: F. nucleatum, F. necrophorum, and F. mortiferum. Adhesion and invasion of the bacteria were assayed before and after antibiotic treatment that killed the adhered and extracellular bacteria. The number of live fusobacteria was examined by bacterial culturing after sonication of the epithelial cells. The role of host cell cytoskeleton functions was examined by treating the epithelial cells with cell function inhibitors. Number of viable epithelial cells was measured with the CellTiter96 kit. The tested Fusobacterium species adhered to and invaded the epithelial cells, and multiplied intracellularly for several hours. Thereafter, the intracellular number of bacteria rapidly declined. Concomitantly, viable fusobacteria were detected in the culture medium. Treatment of the infected epithelial cells with an actin formation inhibitor markedly reduced the number of living intracellular fusobacteria. Newly formed actin filaments were seen by confocal microscopy in the epithelial cells associated with the invaded bacteria. Fusobacteria infection did not reduce the number of viable epithelial cells in culture. Thus, fusobacteria are able to adhere to and invade epithelial cells, and survive under aerobic conditions. This property may enable them to survive in mucosa and participate in various disease processes of oral and pharyngeal tissues.
Serotonin content of normal and inflamed appendix: a possible role of serotonin in acute appendicitis
Authors: VASEI MOHAMMAD , ZAKERI ZEINAB , AZARPIRA NEGAR , HOSSEINI SEYED VAHID , SOLAYMANI‐DODARAN MASOUD
Published: December 2008

The appendix is lined by a mucosa which has many neuroendocrine cells containing serotonin. Local release of serotonin can act as a mediator of inflammation. In this study we explored the serotonin content of the neuroendocrine cells of the appendixes removed for clinical diagnosis of appendicitis. Appendix specimens were divided into three groups: Acute appendicitis (AA), non‐appendicitis (NA), and follicular hyperplasia (FH). Normal appendix specimens from patients undergoing elective abdominal surgery were used as the control group (NL). All sections were exposed to proteinase K, incubated with anti‐serotonin, chromogranin A, and synaptophysin antibodies, and treated with the LSAB kit. Polygonal cells were seen within the crypt epithelium (enterochromaffin cell, EC) and within the lamina propria (subepithelial neuroendocrine cell, SNC). In AA, only 16 cases (64%) showed serotonin staining in non‐destructed glands. There was a significant reduction in the number of ECs in AA compared to the FH (96%), NA (100%) and NL (100%) groups (P<0.001). Chromogranin and synaptophysin immunostaining also showed a significant reduction in the number of ECs in AA compared with the other three groups (P<0.001). SNC serotonin reactivity was lower in the AA group compared with the other groups (p<0.001). The inflamed appendix is markedly depleted of serotonin in the epithelium and lamina propria. Local serotonin release from ECs and SNCs in the appendix may act as an inflammatory mediator in appendicitis and is likely to be the source of raised blood serotonin in AA.
pS2 (TFF1) expression in prostate carcinoma: correlation with steroid receptor status
Authors: ABDOU ASMAA GABER , AIAD HAYAM ABDEL SAMIE , SULTAN SULTAN MOHAMED
Published: December 2008

pS2 or TFF1 is a member of the trefoil factor family, which is distributed throughout the gastrointestinal tract in both normal and diseased tissues. It is also considered to be one of the major estrogen‐regulated proteins and an indicator of estrogen receptor (ER) functionality. pS2 has previously been investigated in benign and malignant prostate lesions with little information about its relationship to steroid receptor status. Our purpose was to correlate pS2 expression with steroid receptor status (ER alpha and progesterone receptor (PR)) and other pathologic variables in prostate carcinoma. 15 benign prostate hyperplasia (BPH) and 47 prostate carcinoma cases were investigated by means of immunohistochemistry for pS2, ER and PR expression. 80% of BPH showed pS2 cytoplasmic immunoreactivity in hyperplastic acini and about half of these cases also exhibited nuclear staining decorating basal or both basal and luminal nuclei. pS2 was highly expressed in prostate carcinoma (91.4%) with both cytoplasmic and nuclear patterns of staining. The latter pattern was significantly associated with carcinoma having a low Gleason score (p=0.02). pS2 lacked any significant correlation with steroid receptor status, stage or grade. Univariate survival analysis revealed a significant impact of stage (p=0.03) and nodal status (p<0.0001) on patient outcome. The diagnostic value of pS2 expression in prostate carcinoma validated 74.19% accuracy, 91.48% sensitivity and 78.18% positive predictive value. The high sensitivity of pS2 expression in prostate carcinoma could make it a suitable marker for diagnosis of prostate carcinoma, especially in metastatic cases of unknown origin. The absence of correlation and dissimilarity in immunolocalization between pS2 and ER alpha leads to the assumption that ER alpha could not be the regulatory protein for pS2 and may raise questions about the functionality of ER alpha in prostate. The nuclear pattern of pS2 immunoreactivity either in benign or malignant prostatic lesions is similar to the published data on ER beta distribution and could also identify a subset of carcinoma patients with a favorable prognosis.
Expression of non‐mast cell histidine decarboxylase in tumor‐associated microvessels in human esophageal squamous cell carcinomas
Authors: LI ZHENFENG , LIU JINZHONG , TANG FUAI , LIU YIQING , WALDUM HELGE L. , CUI GUANGLIN
Published: December 2008

Histamine is produced by mast cells and many other types of cells. The role of histamine released from mast cells in promoting tumor angiogenesis has been intensively studied; however, the role of non‐mast cell histamine in regulating tumor angiogenesis has been largely ignored. In this study, tissue specimen sections from 43 patients with esophageal squamous cell carcinoma (ESCC) and normal esophageal biopsies from 17 heath individuals obtained from a high incidence area of north China were used to assess changes in microvessel density (MVD) and non‐mast cell L‐histidine decarboxylase (HDC) (the only rate‐limiting enzyme that catalyzes the formation of histamine from L‐histidine) expression in the tumor microenvironment by immunohistochemistry (IHC). In addition, the cellular characterization of non‐mast cell HDC‐positive cells in microvessels was examined by double IHC combined with HDC/CD34 and HDC/PCNA antibodies. These IHC analyses revealed a significantly increased HDC‐positive MVD in ESCC as compared with normal controls, which accounted for ∼61% of CD34‐labeled general MVD in ESCC. Furthermore, IHC in serial sections and double IHC showed that most of these HDC‐positive cells were CD34‐positive endothelial cells in microvessels with an increased proliferative capacity. Thus, our results suggest that non‐mast cell histamine expressed in endothelial cells of microvessels could be an additional cellular source and might play a role in regulating angiogenesis in ESCC.
Multiple extragenital adenomatoid tumors in the mesocolon and omentum
Authors: YEH CHI‐JU , CHUANG WEN‐YU , CHOU HUNG‐HSUEH , JUNG SHIH‐MING , HSUEH SWEI
Published: December 2008

Adenomatoid tumors are benign mesothelial neoplasms most commonly found in the male and female genital tracts. Extragenital adenomatoid tumors are rare, most of them being solitary tumors. To our knowledge, only one case of multiple extragenital adenomatoid tumors, involving the liver and peritoneum, has been reported to date. Here we report another case of multiple extragenital adenomatoid tumors involving the mesocolon and omentum. A 47‐year‐old woman presented with a delayed menstrual period. Ultrasonography revealed a left adnexal mass, and surgical resection was attempted at a local hospital. The patient was transferred to our hospital without resection due to the intraoperative finding of multiple peritoneal tumors. At our hospital, an 8.0×7.5×6.0 cm tumor at the mesocolon of the sigmoid colon and three omental nodules measuring up to 2.5×2.0×1.7 cm were resected. Grossly, they were well circumscribed, gray‐tan and elastic with small cystic spaces. Microscopically, they were composed of tubules and anastomosing channels lined by flattened or cuboidal cells with bland nuclei. Immunohistochemically, the tumor cells were positive for pan‐cytokeratin AE1/AE3, vimentin, cytokeratin 5/6 and calretinin. The postoperative course was uneventful, and the patient was well 10 months after the operation. Despite their rarity, adenomatoid tumors should be included in the differential diagnosis of multiple intra‐abdominal tumors.
Immunogenicity and protective efficacy of a DNA vaccine encoding the fusion protein of mycobacterium heat shock protein 65(Hsp65) with human interleukin‐2 against Mycobacterium tuberculosis in BALB/c mice
Authors: WANG LI‐MEI , BAI YIN‐LAN , SHI CHANG‐HONG , GAO HUI , XUE YING , JIANG HONG , XU ZHI‐KAI
Published: December 2008

Developing a new generation of vaccines is important for preventing tuberculosis (TB). DNA vaccine is one promising candidate. In this study we evaluated the immunogenicity and protective efficacy of the DNA vaccine encoding the fusion protein of Mycobacterium tuberculosis heat shock protein 65 (Hsp65) with human interleukin‐2 (hIL‐2) in BALB/c mice. We showed that the DNA vaccine pcDNA‐Hsp65‐hIL‐2 could induce high levels of antigen‐specific antibody, IFN‐γ, CD4+ and CD8+ T cell production. When the immunized mice were infected with M. tuberculosis H37Rv, the organ bacterial loads in the DNA immunized group were significantly reduced compared to those of the saline control group, but the ability to reduce bacteria was not better than for BCG. The histopathology in lungs of the DNA vaccine immunized mice was similar to that of BCG immunized mice, which was obviously ameliorated compared to that of the saline control group. Overall, the DNA vaccine could afford protection against M. tuberculosis infection, though the protection efficacy was not as great as that of conventional BCG.
Enhanced phosphorylation of the epidermal growth factor receptor at the site of tyrosine 992 in esophageal carcinomas
Authors: MIYAWAKI MICHIYO , HIJIYA NAOKI , TSUKAMOTO YOSHIYUKI , NAKADA CHISATO , KAWAHARA KATSUNOBU , MORIYAMA MASATSUGU
Published: December 2008

This study aimed to determine whether epidermal growth factor receptor (EGFR), which has been reported to be frequently overexpressed in esophageal carcinoma cells, is actually activated in the cells. Paraffin‐embedded specimens of 39 cases of esophageal carcinoma were analyzed immunohistochemically with anti‐EGFR polyclonal antibody (α‐EGFR Ab) and also an anti‐phospho‐EGFR‐specific polyclonal antibody (α‐p‐EGFRTyr992 Ab) that specifically recognizes phosphorylated tyrosine 992 of EGFR. All of the 39 cases were found to express EGFR, but the expression levels were not significantly higher than those in basal cells of the normal esophageal epithelium. In 38 of the 39 cases, α‐p‐EGFRTyr992 immunoreactivity was evident. Interestingly, the positively stained carcinoma cells were not distributed diffusely, and strongly immunostained cells tended to be localized in areas of severe dysplasia and in microinvasive foci just adjacent to the main invasive carcinoma. However, the deeply invasive front never exhibited positive immunoreactivity. The present findings suggest that phosphorylation of EGFR Tyr992 may play some specific functional role in esophageal carcinomas besides promotion of cell proliferation.
Scedosporium apiospermum: an emerging opportunistic pathogen that must be distinguished from Aspergillus and other hyalohyphomycetes
Authors: Shinohara Michi M. , George Evan
Published: December 2008

We report a case of cutaneous Scedosporium apiospermum infection in an immunocompromised host. S.apiospermum is an emerging opportunistic pathogen, especially in organ transplant recipients. Prompt identification is critical because of its resistance to most antifungal drugs. Its histopathologic features are indistinct and overlap with those of more commonly recognized hyalohyphomycetes such as Aspergillus species. Cultures from infected tissue are generally required for correct identification. Clinicians and pathologists must be familiar with this organism and recognize the need for culture studies in addition to histopathology in the evaluation of specimens from immunocompromised patients with suspected fungal infection.
Kinetic hybrid models composed of mechanistic and simplified enzymatic rate laws – a promising method for speeding up the kinetic modelling of complex metabolic networks
Authors: Bulik Sascha , Grimbs Sergio , Huthmacher Carola , Selbig Joachim , Holzhütter Hermann G.
Published: December 2008

Kinetic modelling of complex metabolic networks – a central goal of computational systems biology – is currently hampered by the lack of reliable rate equations for the majority of the underlying biochemical reactions and membrane transporters. On the basis of biochemically substantiated evidence that metabolic control is exerted by a narrow set of key regulatory enzymes, we propose here a hybrid modelling approach in which only the central regulatory enzymes are described by detailed mechanistic rate equations, and the majority of enzymes are approximated by simplified (nonmechanistic) rate equations (e.g. mass action, LinLog, Michaelis–Menten and power law) capturing only a few basic kinetic features and hence containing only a small number of parameters to be experimentally determined. To check the reliability of this approach, we have applied it to two different metabolic networks, the energy and redox metabolism of red blood cells, and the purine metabolism of hepatocytes, using in both cases available comprehensive mechanistic models as reference standards. Identification of the central regulatory enzymes was performed by employing only information on network topology and the metabolic data for a single reference state of the network [Grimbs S, Selbig J, Bulik S, Holzhutter HG & Steuer R (2007) Mol Syst Biol3, 146, doi:10.1038/msb4100186]. Calculations of stationary and temporary states under various physiological challenges demonstrate the good performance of the hybrid models. We propose the hybrid modelling approach as a means to speed up the development of reliable kinetic models for complex metabolic networks.
CHILD PROTECTION MEDIATION: A 25‐YEAR PERSPECTIVE
Authors: Edwards Leonard
Published: December 2008

This article reviews the creation, development, and growth of child protection mediation (CPM) in the United States. Starting with a few pilot projects in the 1970s, CPM has grown throughout the country. The article traces child protection's development through the publication of the Resource Guidelines and Model Courts and then discusses what the necessary ingredients for a mediation program are. Mediation is then discussed from a judicial perspective. Barriers to mediation are listed, followed by a discussion of special issues that arise when developing and maintaining CPM programs. The article concludes with the observation that CPM is now recognized as a best practice by most judges and court improvement professionals and that it continues to grow.
CHILD PROTECTION MEDIATION: THE COOK COUNTY ILLINOIS EXPERIENCE—A JUDGE'S PERSPECTIVE
Authors: Martin Patricia M.
Published: December 2008

The Child Protection Mediation Program in Cook County, Illinois is the result of a collaborative effort on the part of the court, its stakeholders, and the attorneys involved in child protection cases. Child protection mediation empowers families and includes parents in many decisions impacting their children. It also helps judges to move children's cases through the legal system more rapidly and in a more efficient and humane manner. This results in children achieving more timely permanency. The program is sustained in part due to frequent outreach to, and input from, the program's consumers. These help to ensure that the program is meeting the diverse needs of the court, the families, and the professionals involved in the cases.
SUPPORTING FAMILY STRENGTH: THE USE OF TRANSFORMATIVE MEDIATION IN A PINS MEDIATION CLINIC
Authors: Baruch Bush Robert A. , Hershman Lisa , Thaler Robert , Vitkovich Christena
Published: December 2008

This article describes, and explains the rationales for, the establishment of an innovative program in which a law school and two social services agencies are collaborating in a mediation clinic that uses the transformative model of mediation to serve clients in cases that seem ideally suited to benefit from this approach to practice—so‐called “PINS” (persons in need of supervision) cases involving conflict within families, between parents and their adolescent children. The article first describes the evolution of the PINS system and the changing views of its purpose, especially in New York State. It then offers a brief introduction to the unique features of the transformative approach to mediation and explains why that approach in particular is likely to mesh with current understandings of the needs of the families involved in these cases. Finally, the article describes the innovative “Family Solutions Program” and how it uses transformative mediation to help these families and suggests that experience of this program shows how use of the transformative model, in particular, could be valuable not only for clinical education but also for effective volunteer training and supervision in mediation programs generally.
OUT‐OF‐COURT STATEMENTS BY VICTIMS OF CHILD SEXUAL ABUSE TO MULTIDISCIPLINARY TEAMS: A CONFRONTATION CLAUSE ANALYSIS
Authors: Scher Jonathan
Published: December 2008

Acknowledging the rapid growth of child sexual abuse in the United States, this Note advocates for the recognition of a limited exception to the blanket‐hearsay ban on out‐of‐court statements made by unavailable declarants set out by the Supreme Court in Crawford v. Washington. In order to protect a criminal defendant's Sixth Amendment confrontation right, Crawford requires that hearsay evidence that is “testimonial” in nature be deemed inadmissible if the witness is unavailable and the defendant does not have a prior opportunity to cross‐examine the witness against him. However, Crawford noted that, where nontestimonial hearsay is at issue, cross‐examination may not be necessary. Accordingly, where a child sexual abuse victim makes statements during a structured or semi‐structured forensic interview to a member of a multidisciplinary team, these statements should be deemed nontestimonial and thus admitted into evidence, without requiring cross‐examination of the child. Allowing for this exception to the general hearsay ban in Crawford is not only consistent with current precedent, but it is also warranted to promote public policy and to curb the negative impact such abuse has on society.
Characters of Cysteine Endopeptidases in Wheat Endosperm during Seed Germination and Subsequent Seedling Growth
Authors: Shi Chao , Xu Lang‐Lai
Published: December 2008

The endopeptidases (EPs) in wheat endosperm during seed germination and subsequent seedling growth were characterized by gradient‐polyacrylamide gel electrophoresis with gelatin copolymerized into the gel. Four cysteine EPs (EP1, EP2, EP3 and EP4) were detected in wheat endosperm during the 7 d growth after seed imbibition. The results also showed that the activities of all of these EPs increased continuously, and EP2 first appeared and had the highest proteolytic activity among the four EPs in this experimental process. The optimum pH and temperature of all four EPs were 4.0 and 40.0 °C. All EPs were completely inhibited by 25 μmol/L E‐64 and had no good thermal stabilities, especially EP1. In addition, these EPs had different substrate specificities to albumins, globulins, gliadins and glutenins; the main storage proteins of mature wheat endosperm. Among them, EP2 had the highest proteolytic activities on globulins, gliadins and glutenins, and might be the most important and specific EP with potential to be tightly correlated with seedling development.