bluuebunny/crossref_metadata_2025 huggingface dataset:
T17b murine embryonal endothelial progenitor cells can be induced towards both proliferation and differentiation in a fibrin matrix
Authors: Bleiziffer Oliver , Horch Raymund E. , Hammon Matthias , Arkudas Andreas , Naschberger Elisabeth , Rath Subha , Pryymachuk Galyna , Beier Justus P. , Hatzopoulos Antonis K. , Stürzl Michael , Kneser Ulrich
Published: October 2008

Endothelial progenitor cells (EPC) may enhance blood vessel formation in a variety of clinical settings such as ischaemia and tumour angiogenesis as well as in tissue‐engineered matrices. In the present study, we cultured a murine endothelial progenitor cell line, T17b, in vitro in cell culture as well as in an FDA‐approved fibrin matrix and investigated cell proliferation, differentiation and secretion patterns of the angiogenic growth factor VEGF under hypoxia and differentiation. We show that T17b EPC remain viable for at least 8 days in the fibrin matrix where they proliferate and form clusters including lumen‐like structures. Proliferation in fibrin clots overlayed with basal medium (BM) was confirmed morphologically and immunohistochemically by positive Ki67 staining, indicating mitotic activity. Significant cell proliferation and Ki‐67 expression were absent when cells were incubated with dibutyryl‐cAMP and retinoic acid (RA). Incubation with dibutyryl‐cAMP and RA stimulated the expression of the EPC differentiation markers von Willebrand Factor (vWF) and VEGF receptor 2 (VEGFR‐2), indicating successful differentiation in the fibrin clot. EPC differentiation induced by dibutyryl‐cAMP and RA was confirmed in 2‐D chamber slide cultures by positive vWF immunostaining, which was absent in BM controls. EPC chamber slides also displayed positive vWF staining when exposed to hypoxia under BM conditions, indicating EPC activation and differentiation could also be induced by hypoxia. Taken together, T17b EPC secrete increased levels of VEGF when submitted to either hypoxia or differentiation and can be differentiated into mature endothelial cells not only in cell and matrigel cultures but also in a fibrin matrix that is FDA approved for clinical application.
Reduced ligand affinity leads to an impaired function of the adenosine A2A receptor of human granulocytes in sepsis
Authors: Kreth Simone , Kaufmann Ines , Ledderose Carola , Luchting Benjamin , Thiel Manfred
Published: October 2008

The enhanced release of reactive oxygen species by excessively activated polymorphonuclear leucocytes (PMN) is a key step in the pathogenesis of sepsis. Potent action of adenosine in inhibiting cytotoxic PMN functions has been documented. Recent data, however provide evidence that in sepsis a diminished capability of adenosine to inhibit the generation of oxygen radicals by PMN occurs. Here, we investigated the underlying mechanisms in an in vitro sepsis model and in PMN of sepsis patients. We report that lipopolysaccharide (LPS)‐incubation of human PMN elicited the same increase in the half‐maximal inhibitory concentration (IC50) of adenosine as observed in patients with septic shock. Coupling to adenylyl cyclase was impaired as well, as indicated by a decreased potency of adenosine to stimulate cyclic adenosine monophosphate (cAMP) accumulation. Ligand‐binding studies conducted with native, LPS‐stimulated PMN, and with PMN of sepsis patients revealed that, despite an increased adenosine A2A receptor (A2AR) expression, the receptor function declines due to a diminished ligand‐binding affinity most likely caused by allosteric modulators within the inflammatory environment. A2AR function obviously is highly dependent upon the cellular environment and thus, further functional characterization of A2AR responses in sepsis may be a promising approach to develop new adenosine or A2AR agonists based therapeutic strategies.
A HCMV pp65 polypeptide promotes the expansion of CD4+ and CD8+ T cells across a wide range of HLA specificities
Authors: Provenzano Maurizio , Sais Giovanni , Bracci Laura , Egli Adrian , Anselmi Maurizio , Viehl Carsten T , Schaub Stefan , Hirsch Hans H , Stroncek David F , Marincola Francesco M. , Spagnoli Giulio C
Published: October 2008

Human cytomegalovirus (HCMV) can cause life‐threatening disease in infected hosts. Immunization with human leukocyte antigen (HLA)‐restricted immunodominant synthetic peptides and adoptive transfer of epitope‐specific T cells have been envisaged to generate or boost HCMV‐specific cellular immunity, thereby preventing HCMV infection or reactivation. However, induction or expansion of T cells effective against HCMV are limited by the need of utilizing peptides with defined HLA restrictions. We took advantage of a combination of seven predictive algorithms to identify immunogenic peptides of potential use in the prevention or treatment of HCMV infection or reactivation. Here we describe a pp65‐derived peptide (pp65340–355, RQYDPVAALFFFDIDL: RQY16‐mer), characterized by peculiar features. First, RQY‐16mer is able to stimulate HCMV pp65 specific responses in both CD4+ and CD8+ T cells, restricted by a wide range of HLA class I and II determinants. Second, RQY‐16mer is able to induce an unusually wide range of effector functions in CD4+ T cells, including proliferation, killing of autologous HCMV‐infected target cells and cytokine production. Third, and most importantly, the RQY‐16mer is able to stimulate CD4+ and CD8+ T‐cell responses in pharmacologically immunosuppressed patients. These data suggest that a single reagent might qualify as synthetic immunogen for potentially large populations exposed to HCMV infection or reactivation.
Drug releasing systems in cardiovascular tissue engineering
Authors: Spadaccio Cristiano , Chello Massimo , Trombetta Marcella , Rainer Alberto , Toyoda Yoshiya , Genovese Jorge A.
Published: October 2008

Introduction

Drug and molecules released

Conclusion

Acknowledgements

Heart disease and atherosclerosis are the leading causes of morbidity and mortality worldwide. The lack of suitable autologous grafts has produced a need for artificial grafts; however, current artificial grafts carry significant limitations, including thrombosis, infection, limited durability and the inability to grow. Tissue engineering of blood vessels, cardiovascular structures and whole organs is a promising approach for creating replacement tissues to repair congenital defects and/or diseased tissues. In an attempt to surmount the shortcomings of artificial grafts, tissue‐engineered cardiovascular graft (TECVG), constructs obtained using cultured autologous vascular cells seeded onto a synthetic biodegradable polymer scaffold, have been developed. Autologous TECVGs have the potential advantages of growth, durability, resistance to infection, and freedom from problems of rejection, thrombogenicity and donor scarcity. Moreover polymers engrafted with growth factors, cytokines, drugs have been developed allowing drug‐releasing systems capable of focused and localized delivery of molecules depending on the environmental requirements and the milieu in which the scaffold is placed. A broad range of applications for compound‐releasing, tissue‐engineered grafts have been suggested ranging from drug delivery to gene therapy. This review will describe advances in the development of drug‐delivery systems for cardiovascular applications focusing on the manufacturing techniques and on the compounds delivered by these systems to date.
Advances in microRNAs: implications for immunity and inflammatory diseases
Authors: Sonkoly E. , Pivarcsi A.
Published: October 2008

Introduction

Identification of microRNA targets

Multi‐level control of microRNA expression

How can microRNAs contribute to diseases?

MiRNAs in immunological and inflammatory disorders

MiRNAs in antiviral immunity and viral immune escape

Perspectives

Since their discovery in 1993 and the introduction of the termmicroRNAin 2001, it has become evident that microRNAs (miRNAs) involved in many biological processes, including development, differentiation, proliferation and apoptosis. The function of miRNA the control of protein production in cells by sequence‐specific targeting of mRNAs for translational repression or mRNA degradati Interestingly, immune genes are apparently preferentially targeted by miRNAs compared to the average of the human genome, indicat the significance of miRNA‐mediated regulation for normal immune responses. Here, we review what is known about the role of miRN in the pathogenesis of immune‐related diseases such as chronic inflammatory skin diseases, autoimmunity and viral infections.
Epigenetic programming of μ‐opioid receptor gene in mouse brain is regulated by MeCP2 and brg1 chromatin remodelling factor
Authors: Hwang Cheol Kyu , Song Kyu Young , Kim Chun Sung , Choi Hack Sun , Guo Xiao‐Hong , Law Ping‐Yee , Wei Li‐Na , Loh Horace H.
Published: October 2008

The pharmacological action of morphine as a pain medication is mediated primarily through the μ‐opioid receptor (MOR). With few exceptions, MOR is expressed in brain regions where opioid actions take place. The basis for this unique spatial expression of MOR remains undetermined. Recently, we reported that DNA methylation of the MOR promoter plays an important role in regulating MOR in P19 cells. In this study, we show that the differential expression of MOR in microdissected mouse brain regions coincides with DNA methylation and histone modifications. MOR expression could be induced by a demethylating agent or a histone deacetylase inhibitor in MOR‐negative cells, suggesting that the MOR gene can be silenced under epigenetic control. Increases in the in vivo interaction of methyl‐CpG‐binding protein 2 (MeCP2) were observed in the cerebellum, in which the MOR promoter was hypermethylated and MOR expression was the lowest among all brain regions tested. MeCP2 is associated closely with Rett syndrome, a neurodevelopmental disorder. We also established novel evidence for a functional role for MeCP2’s association with the chromatin‐remodelling factor Brg1 and DNA methyltransferase Dnmt1, suggesting a possible role for MeCP2 in chromatin remodelling during MOR gene regulation. We conclude that MOR gene expression is epigenetically programmed in various brain regions and that MeCP2 assists the epigenetic program during DNA methylation and chromatin remodelling of the MOR promoter.
The potential role of MLC phosphatase and MAPK signalling in the pathogenesis of vascular dysfunction in heart failure
Authors: Ogut Ozgur , Brozovich Frank V.
Published: October 2008

Clinical syndrome of heart failure

Regulation of smooth muscle contractility

Ca2+ sensitization

Ca2+ desensitization

MYPT1 isoforms and the sensitivity to cGMP

cGMP and MYPT1 phosphorylation

Vascular function and cGMP signalling

Captopril therapy and MYPT1 Expression in HF

Captopril therapy and gene expression

Conclusions

The clinical syndrome of heart failure is associated with both a resting vasoconstriction and reduced sensitivity to nitric oxide mediated vasodilatation, and this review will focus on the role of myosin light chain (MLC) phosphatase in the pathogenesis of the vascular abnormalities of heart failure. Nitric oxide mediates vasodilatation by an activation of guanylate cyclase and an increase in the production of cGMP, which leads to the activation of the type I cGMP‐dependent protein kinase (PKGI). PKGI then activates a number of targets that produce smooth muscle relaxation including MLC phosphatase. MLC phosphatase is a holoenzyme consisting of three subunits; a 20 kD subunit of unknown function, an ∼38‐kD catalytic subunit and a myosin targeting subunit (MYPT1). Alternative splicing of a 31 bp 3' exon generates MYPT1 isoforms, which differ by a COOH‐terminus leucine zipper (LZ). Further, PKGI‐mediated activation of MLC phosphatase requires the expression of a LZ+ MYPT1. Congestive heart failure is associated with a decrease in LZ+ MYPT1 expression, which results in a decrease in the sensitivity to cGMP‐mediated smooth muscle relaxation. Beyond their ability to reduce afterload, angiotensin converting enzyme (ACE) inhibitors have a number of beneficial effects that include maintaining the expression of the LZ+ MYPT1 isoform, thereby conserving normal sensitivity to cGMP‐mediated vasodilatation, as well as differentially regulating genes associated with mitogen activated protein kinase (MAPK) signalling. ACE inhibition reduces circulating angiotensin II and thus limits the downstream activation of MAPK signalling pathways, possibly preventing the alteration of the vascular phenotype to preserve normal vascular function.
Intraplaque injection of Ad5‐CMV.p53 aggravates local inflammation and leads to plaque instability in rabbits
Authors: Zhang Lei , Liu Yan , Lu Xiao Ting , Xu Xin Sheng , Zhao Yu Xia , Ji Xiao Ping , Zhang Peng Fei , Liu Chun Xi , Tang Meng Xiong , Chen Wen Qiang , Zhang Yun
Published: October 2008

This study aims to develop a new animal model of vulnerable plaques and investigate the potential mechanisms of exogenous p53‐induced plaque instability. Forty rabbits underwent aortic balloon injury, were fed a 1% cholesterol diet for 10 weeks and then normal chow for 6 weeks. Rabbits were divided into Ad5‐CMV.p53‐treated group (n= 16), Ad5‐CMV.lac Z‐treated group (n= 16) and blank control group (n= 8). Under the guidance of intravascular ultrasound, a 50‐μl suspension of adenovirus containing p53 or lac Z was injected into the largest plaque of the first two groups, respectively, and these rabbits received pharmacological triggering 2 weeks later. In 76.9% of rabbits with p53 transfection, plaque rupture was found, which was significantly (P < 0.05) higher than that in the Ad5‐CMV.lac Z‐treated plaques (23.1%), or blank controls plaques (0%). Increased apoptotic cells, and subsequently, decreased vascular smooth muscle cells and collagen content, enhanced intima macrophage accumulation, increased C‐reactive protein (CRP) and matrix metalloproteinases staining and high serum levels of high sensitive CRP (hs‐CRP) and monocyte chemoattractant protein‐1 (MCP‐1) were observed in Ad5‐CMV.p53‐treated rabbits. However, a binary logistic regression model revealed that hs‐CRP concentration rather than apoptosis rate played an independent role in plaque rupture with an odds ratio as 1.314 (95% CI: 1.041–1.657, P= 0.021), and there were high positive correlations between inflammatory biomarkers (hs‐CRP or MCP‐1) and apoptosis (R2= 0.761, and R2= 0.557, respectively, both P < 0.01). Intraplaque injection of p53 gene provides a safe and effective method for inducing plaque vulnerability in rabbits. The destabilizing effect of p53 overexpression is mediated mainly through apoptosis‐enhanced inflammation rather than cell apoptosis itself.
Combination of taxol and Bcl‐2 siRNA induces apoptosis in human glioblastoma cells and inhibits invasion, angiogenesis and tumour growth
Authors: George Joseph , Banik Naren L. , Ray Swapan K.
Published: October 2008

Taxol is a powerful chemotherapeutic agent that binds to microtubules to prevent tumour cell division. However, a traditional high dose of taxol may also induce apoptosis in normal cells. The anti‐apoptotic molecule Bcl‐2 is up‐regulated in tumour cells to prevent apoptosis. We designed this study to determine whether use of a low dose of taxol and anti‐apoptotic Bcl‐2 gene silencing would effectively induce apoptosis in human glioblastoma U251MG cells and also inhibit invasion, angiogenesis and intracranial as well as subcutaneous tumour growth. We treated the cells with either 100 nM taxol or transfected with a plasmid vector expressing Bcl‐2 siRNA or both agents together for 72 h. Knockdown of Bcl‐2 potentiated efficacy of taxol for cell death. Fluorescence‐activated cell sorting analysis, double immunofluorescent staining and TUNEL assay demonstrated apoptosis in about 70% of the cells after treatment with the combination of taxol and Bcl‐2 siRNA. In vitro Matrigel invasion assay demonstrated dramatic decrease in glioblastoma cell invasion and in vivo angiogenesis assay showed complete inhibition of neovascularization in athymic nude mice after treatment with the combination. Further, treatment with the combination of taxol and Bcl‐2 siRNA caused suppression of intracranial tumour growth and subcutaneous solid tumour development. In conclusion, our results indicate that the combination of taxol and Bcl‐2 siRNA effectively induces apoptosis and inhibits glioblastoma cell invasion, angiogenesis and intracranial as well as subcutaneous tumour growth. Therefore, the combination of a low dose of taxol and Bcl‐2 siRNA is a promising therapeutic strategy for controlling the aggressive growth of human glioblastoma.
Let‐7 repression leads to HMGA2 overexpression in uterine leiomyosarcoma
Authors: Shi Guizhi , Perle Mary Ann , Mittal Khush , Chen Hua , Zou Xuanyi , Narita Masashi , Hernando Eva , Lee Peng , Wei Jian‐Jun
Published: October 2008

Overexpression of HMGA2 is common in uterine leiomyomas (ULM). The expression of HMGA2 in its malignant counterpart – uterine leiomyosarcomas (ULMS) remains undetermined. Recently it has been shown that repression of HMGA2 by microRNA let‐7s is a critical molecular regulatory mechanism associated with tumour growth in many tumours and cell types, including leiomyomas. To test whether HMGA2 and let‐7s play a role in ULMS, we examined the levels of endogenous HMGA2 and let‐7 expression and found a significant correlation between these two molecules in a case‐matched cohort of human ULMS. We found that overexpression of HMGA2 and let‐7‐mediated HMGA2 repression is a relevant molecular alteration in ULMS. Disrupting the control of HMGA2 and let‐7 pairs promotes ULMS cell growth in vitro.
The missing link between indoleamine 2,3‐dioxygenase mediated antibacterial and immunoregulatory effects
Authors: Müller Anika , Heseler Kathrin , Schmidt Silvia K. , Spekker Katrin , MacKenzie Colin R. , Däubener Walter
Published: October 2008

The interferon (IFN)–γ‐inducible tryptophan degrading enzyme indoleamine 2,3‐dioxygenase (IDO) has not only been recognized as a potent antimicrobial effector molecule for the last 25 years but was recently found also to have potent immunoregulatory properties. In this study, we provide evidence that both tryptophan starvation and production of toxic tryptophan metabolites are involved in the immunoregulation mediated by IDO, whereas tryptophan starvation seems to be the only antibacterial effector mechanism. A long‐studied controversy in the IDO research field is the seemingly contradictory effect of IDO in the defence against infectious diseases. On the one hand, IFN‐γ‐induced IDO activity mediates an antimicrobial effect, while at the same time IDO inhibits T‐cell proliferation and IFN–γ production. Here, we suggest that both effects, dependent on the threshold for tryptophan, cooperate in a reasonable coherence. We found that the minimum concentration of tryptophan required for bacterial growth is 10‐40‐fold higher than the minimum concentration necessary for T‐cell activation. Therefore, we suggest that during the first phase of infection the IDO‐mediated tryptophan depletion has a predominantly antimicrobial effect whereas in the next stage, and with ongoing tryptophan degradation, the minimum threshold concentration of tryptophan for T‐cell activation is undercut, resulting in an inhibition of T‐cell growth and subsequent IDO activation.
Statin‐induced calcification in human mesenchymal stem cells is cell death related
Authors: Kupcsik Laszlo , Meurya Thomas , Flury Matthias , Stoddart Martin , Alini Mauro
Published: October 2008

Statins are widely used in clinics to lower cholesterol levels. Recently, they have been shown to positively affect bone formation and bone mass in a rat model. The aim of this study was to investigate the effect of pravastatin, simvastatin and lovastatin on the osteoblastic differentiation of human mesenchymal stem cells (MSCs)in vitro. Cell number, alkaline phosphatase (ALP) activity, matrix mineralization and gene expression pattern were determined. Pravastatin did not affect cell differentiation. Simvastatin and lovastatin enhanced bone morphogenetic protein 2 (BMP‐2) mRNA levels. In contrast, ALP activity and mRNA levels were suppressed by statins, as well as the DNA content and cell activity (MTT). An increase in apoptotic events was observed at high concentrations of statins, along with high Ca‐45 incorporation. Lower concentrations of statins did not increase apoptotic staining, but also failed to induce calcification. When statin‐induced calcification did occur, the morphology of the deposits was very different from the conventional nodule formation; the calcium was laid down along the membranes of the rounded cells suggesting it was as a result of cell death. Our results indicate that statins are not able to differentiate human MSCs into osteoblasts and that high concentrations of statins (>1 μM) have a cytotoxic effect.
Retracted:Advanced glycation endproduct (AGE) accumulation and AGE receptor (RAGE) up‐regulation contribute to the onset of diabetic cardiomyopathy
Authors: Ma Heng , Li Shi‐Yan , Xu Peisheng , Babcock Sara A. , Dolence E. Kurt , Brownlee Michael , Li Ji , Ren Jun
Published: October 2008

Diabetic cardiomyopathy is manifested by compromised systolic and diastolic function. This study was designed to examine the role of advanced glycation endproduct (AGE) and AGE receptor (RAGE) in diabetic cardiomyopathy. Heart function was assessed in isolated control and streptozotocin‐induced diabetic hearts followingin vivoRAGE gene knockdown using RNA interference. Cardiomyocyte mechanical properties were evaluated including peak shortening (PS), time‐to‐PS (TPS) and time‐to‐90% relengthening (TR90). RAGE was assayed by RT‐PCR and immunoblot. Diabetes significantly enhanced cardiac MG, AGE and RAGE levels accompanied with colocalization of AGE and RAGE in cardiomyocytes. Diabetes‐elicited increase in RAGE was inhibited byin vivosiRNA interference. The AGE formation inhibitor benfotiamine significantly attenuated diabetes‐induced elevation in MG, AGE, RAGE and collagen cross‐linking without affecting hypertriglyceridaemia and hypercholesterolaemia in diabetes. Diabetes markedly decreased LV contractility, as evidenced by reduced ±dP/dt and LV developed pressure (LVDP), which were protected by RAGE gene knockdown. In addition, MG‐derived AGE (MG‐AGE) up‐regulated cardiac RAGE mRNA and triggered cardiomyocyte contractile dysfunction reminiscent of diabetic cardiomyopathy. The MG‐AGE‐elicited prolongation of TPS and TR90was ablated by an anti‐RAGE antibody in cardiomyocytes. Interestingly, MG‐AGE‐induced cardiomyocyte dysfunction was associated with mitochondrial membrane potential (MMP) depolarization and reduced GSK‐3β inactivation in control cardiomyocytes, similar to those fromin vivodiabetes. Treatment with siRNA‐RAGE ablated diabetes‐induced MMP depolarization and GSK‐3β inactivation. Collectively, our result implicated a role of AGE‐RAGE in the pathogenesis of diabetic cardiomyopathy.
E10A, an adenovirus carrying human endostatin gene, in combination with docetaxel treatment inhibits prostate cancer growth and metastases
Authors: Zhao Peng , Luo Rongcheng , Wu Jiangxue , Xie Fajun , Li Hongli , Xiao Xia , Fu Liwu , Zhu Xiaofeng , Liu Ranyi , Zhu Yinghui , Liang Zhihui , Huang Wenlin
Published: October 2008

E10A, a replication‐defective adenovirus carrying human endostatin gene, has finished Phase I clinical trials for solid cancers. We assessed whether the combination of E10A with docetaxel would enhance antiangiogenic activities and inhibit prostate cancer growth and metastases. Combination use of conditioned medium from prostate cancer cells infected by E10A and docetaxel exerted synergistic inhibition of HUVECs proliferation, migration and tube formation, compared with either agent alone. In prostate cancer s.c. xenograft models, combined therapy resulted in significant tumour growth inhibition and survival improvement. The antitumoural effect was tightly correlated with a remarkable decrease in tumour cell proliferation, microvessel, especially immature vasculature and significant increase in apoptosis induction. Systemic administration of E10A and docetaxel also effectively inhibited orthotopic growth and metastases of prostate cancer and achieved better in vivo antiangiogenic effects than either agent alone. Our data indicate that E10A in combination with docetaxel exert enhanced antiangiogenic activities and inhibit prostate cancer growth and metastases. Therefore, this approach may be an effective treatment for advanced prostate cancer and deserves more extensive investigation.
Transcriptional regulation of survivin by c‐Myc in BCR/ABL‐transformed cells: implications in anti‐leukaemic strategy
Authors: Fang Zhi Hong , Dong Chun Lan , Chen Zhong , Zhou Bin , Liu Na , Lan Hai Feng , Liang Lu , Liao Wen Bin , Zhang Lei , Han Zhong Chao
Published: October 2008

BCR/ABL can cause chronic myelogenous leukaemia (CML) in part by altering the transcription of specific genes with growth‐ and/or survival‐promoting functions. Recently, BCR/ABL has been shown to activate survivin, an important regulator of cell growth and survival, but the precise molecular mechanisms behind its expression and consequences thereof in CML cells remain unclear. Here, we reported that BCR/ABL promotes survivin expression and its cytoplasmic accumulation. The increase of survivin was largely controlled at the transcriptional level through a mechanism mediated by JAK2/PI3K signal pathways that activated c‐Myc, leading to transactivation of survivin promoter. Dynamic down‐regulation of survivin was a key event involved in imatinib‐induced cell death while forced expression of survivin partially counteracted imatinib's effect on cell survival. Additionally, shRNA‐mediated silencing of survivin or c‐Myc eradicated colony formation of K562 cells in semi‐solid culture system, implying an essential role for this transcriptional network in BCR/ABL‐mediated cell transformation and survival. Finally, interruption of c‐Myc activity by 10058‐F4 exerted an anti‐leukaemia effect with a synergistic interaction with imatinib and overcame the anti‐apoptosis rescued by IL‐3 supplement. In conclusion, we have identified JAK2/PI3K‐mediated and c‐Myc‐dependent transactivation of survivin as a novel pathway in the transcriptional network orchestrated by BCR/ABL. These results suggest that the interference with this circuitry might be a potential utility for CML treatment.
Ginkgolide B preconditioning protects neurons against ischaemia‐induced apoptosis
Authors: Wu Xiaomei , Qian Zhongming , Ke Ya , Du Fang , Zhu Li
Published: October 2008

Ischaemic preconditioning (IP) has been reported to protect the brain against subsequent lethal ischaemia, but it has not been used clinically to prevent ischaemic injury because of safety concerns. The aim of the present study was to see whether Ginkgolide B (GB) is capable of preconditioning as IP to protect neurons against ischaemic injury; if so, which mechanism is involved. Cultured mouse cortical neurons at day 8 were pre‐treated with GB (120 μmol/l) for 24 hrs or exposed to short‐term ischaemia (1 hr) followed by 24‐hr normal culture to induce IP before being treated with severe ischaemia (5 hrs). GB and IP significantly increased cell viability, expression of hypoxia‐inducible factor‐1 alpha (HIF‐1α), erythropoietin (EPO), phosphorylated Bad at serine 136 (136p‐Bad) and phosphorylated glycogen synthase kinase‐ 3β at serine 9 (p‐GSK‐3β), and decreased the percentage of apoptotic cells and the level of active caspase‐3 in severely ischaemic neurons. Moreover, LY294002 that is a specific inhibitor of phosphatidylinositol 3‐kinase (PI3K) significantly reduced the enhanced expression of HIF‐1α, EPO and 136p‐Bad induced by GB and IP. These results suggest that GB, like IP in neurons, is capable of preconditioning against ischaemia‐induced apoptosis, the mechanism of which may involve the PI3K signalling pathway.
FAY‐NE15 — Another Neolithic graveyard in the Central Region of the Sharjah Emirate?
Authors: Kutterer Adelina U. , De Beauclair Roland
Published: October 2008

The paper describes indications for the existence of another substantial Neolithic graveyard of the fifth millennium BC besides al‐Buhais 18 (BHS18) in the Central Region of the Sharjah Emirate. Archaeological explorations in 2006 recovered skeletal remains of three individuals from a primary burial. Deposition of the deceased was similar to BHS18. All three were richly adorned with different kinds of beads found in the head and neck areas. A total of 949 finds of beads from FAY‐NE15 are compared to the large complex of personal adornments from BHS18. There are slight deviations that might indicate that the two graveyards were used by different groups. Implications for the relations between the groups are discussed.
An Umm an‐Nar‐type compartmented soft‐stone vessel from Gonur Depe, Turkmenistan
Authors: Potts D.T.
Published: October 2008

Recent excavations by V.I. Sarianidi in the so‐called ‘royal necropolis’ of Gonur Depe North, in southern Turkmenistan, have brought to light the first unequivocal example of an Umm an‐Nar‐type soft‐stone compartmented vessel ever found in Central Asia. The find is discussed in light of other evidence of links between Central Asia, south‐eastern Arabia and Bahrain.
Late third‐millennium elite burials in Bahrain
Authors: Højlund Flemming , Hilton Anna S. , Juel Christian , Kirkeby Nanna , Laursen Steffen T. , Nielsen Lars E.
Published: October 2008

Studies of old aerial photographs of the Bahrain burial mound fields have revealed that a small number of both Early Type (c. 2200–2050 BC) and Late Type (c. 2050–1750 BC) mounds are encircled by an outer ring wall, apparently marking out the mound as belonging to an elite. Four of these mounds have been excavated, and the results are presented. The geological differences between the Early Type and the Late Type mound landscapes are discussed.
Kinship terms in the Nabataean inscriptions
Authors: Al‐Qudrah Hussein , Abdelaziz Mahdi
Published: October 2008

This paper examines one of the most important aspects of Nabataean social structure, namely kinship terminology. It examines the linguistic derivations, meanings and social significance of kinship terms, and compares them with those found in other Semitic languages.
Gut hormones and appetite control
Authors: Hameed S , Dhillo WS , Bloom SR
Published: October 2008

The gastrointestinal tract is the largest endocrine organ in the body. It secretes more than 20 different peptide hormones, which serve both a local regulatory function and provide a means by which the gut can regulate appetite and satiety. As the worldwide prevalence of obesity reaches epidemic proportions, the importance of delineating the mechanisms which regulate food intake becomes even more urgent. There is now a substantial body of work in both rodent and human models demonstrating the effects of these peptides on appetite and work is underway to therapeutically manipulate the gut‐brain axis for the treatment of obesity. In addition, it may also be possible to use our understanding of the entero‐endocrine system to treat calorie‐deficient states.
Recent aspects of medical care of malignant melanoma
Authors: Terheyden Patrick , Tilgen Wolfgang , Hauschild Axel
Published: October 2008

Recent developments in the epidemiology, diagnosis and therapy of malignant melanoma are reviewed, with particular attention paid to established standards of care. When melanoma metastases are inoperable, they respond poorly to the various chemotherapy strategies, so that additional improvements are critically needed. Cytotoxic T‐lymphocyte antigen‐4 antibodies, multikinase inhibitors, anti‐apoptotic strategies and several other approaches are in progress in Phase III trials both as monotherapy as well as in combination with standard chemotherapy.
The effect of fermented apple diet supplementation on the growth performance and meat quality in finishing pigs
Authors: LEE Sung Dae , KIM Hoi Yun , JUNG Hyun Jung , JI Sang Yun , CHOWDAPPA Rekha , HA Ji Hee , SONG Young Min , PARK Jun Cheol , MOON Hong Kil , KIM In Cheul
Published: October 2008

The objective of the present study was to investigate the effect of fermented apple diet (FAD) supplementation on the growth performance and meat quality in finishing Berkshires. The FAD was made from dropped apple mixed with rice bran and barley bran. Until 81 ± 1 kg live weight at 133 ± 1 days, the animals were fed a growing diet, after which experimental samples were fixed at 0, 2, 4 and 6% FAD as C, T1, T2 and T3 in the finishing diets. Growth performance, ADG, ADFI and feed efficiency were improved in T1 than other groups. In carcass parameters, carcass weight was higher (P < 0.05) in T1 than in other groups. In meat quality, moisture and crude protein contents decreased (P < 0.05) by addition of FAD. pH24 and WHC were higher (P < 0.05) in T1 than other groups. In sensory evaluation, marbling of fresh meat and tenderness, juiciness, flavor and overall acceptability of cooked meat were improved by the addition of FAD. According to the results of our experiment, FAD can be used for improvement of meat quality parameters.
Relationships between physical and structural properties of intramuscular connective tissue and toughness of raw pork
Authors: NISHIMURA Takanori , FANG Suhong , WAKAMATSU Jun‐ichi , TAKAHASHI Koui
Published: October 2008

We studied the relationships between the shear‐force value and physical and structural properties of the intramuscular connective tissue (IMCT) in six classes of porcine skeletal muscle to elucidate the contribution of IMCT to toughness of raw pork. The shear‐force value of raw pork correlated significantly with that of the IMCT model prepared from each class of skeletal muscle (P < 0.05). The correlation suggested that the variable toughness of pork was caused by the mechanical strength of the endomysium and perimysium. The thickness of the secondary perimysium correlated significantly with the shear‐force value of raw pork (P < 0.05) and with that of the IMCT model (P < 0.05). The shear‐force value of raw pork correlated significantly with the total amount of collagen (P < 0.05) but not with the heat‐solubility of collagen. We concluded therefore that the thickness of the secondary perimysium determines the mechanical strength of IMCT and contributes to toughness in raw pork.
The evaluation of the curd forming ability of milk replacers
Authors: OKADA Keiji , KATO Junro , MIYAZAKI Tamako , IKUTA Kentaro , NAITO Yoshihisa , YASUDA Jun
Published: October 2008

Inadequate milk curd formation in the abomasum of newborn calves causes malnutrition and diarrhea. In order to define the factors of inadequate curd formation, we compared the curd forming ability among 9 kinds of milk replacers, bulk milk (raw milk), and skim milk both in vitro and in vivo. When rennet was added, the raw milk and one milk replacer formed firm curds. The rest of the milk replacers and skim milk did not form any curd. When a solution of HCl was added, raw milk, three milk replacers and skim milk formed the curd at pH 4.5, but the other milk replacers did not. When HCl was added following the rennet, raw milk, one milk replacer and skim milk formed the curd. In vivo, raw milk, two milk replacers and skim milk showed good curd formation whereas the other milk replacers showed poor curd formation inside the abomasums of the calves. This study showed that most of the milk replacers sold in Japan could not form the curd with rennet.
Energetic and metabolic transient response of Saccharomyces cerevisiae to benzoic acid
Authors: Kresnowati M. T. A. P. , van Winden W. A. , van Gulik W. M. , Heijnen J. J.
Published: October 2008

Saccharomyces cerevisiae is known to be able to adapt to the presence of the commonly used food preservative benzoic acid with a large energy expenditure. Some mechanisms for the adaptation process have been suggested, but its quantitative energetic and metabolic aspects have rarely been discussed. This study discusses use of the stimulus response approach to quantitatively study the energetic and metabolic aspects of the transient adaptation of S. cerevisiae to a shift in benzoic acid concentration, from 0 to 0.8 mm. The information obtained also serves as the basis for further utilization of benzoic acid as a tool for targeted perturbation of the energy system, which is important in studying the kinetics and regulation of central carbon metabolism in S. cerevisiae. Using this experimental set‐up, we found significant fast‐transient (< 3000 s) increases in O2 consumption and CO2 production rates, of ∼ 50%, which reflect a high energy requirement for the adaptation process. We also found that with a longer exposure time to benzoic acid, S. cerevisiae decreases the cell membrane permeability for this weak acid by a factor of 10 and decreases the cell size to ∼ 80% of the initial value. The intracellular metabolite profile in the new steady‐state indicates increases in the glycolytic and tricarboxylic acid cycle fluxes, which are in agreement with the observed increases in specific glucose and O2 uptake rates.
Microarray analyses of hypoxia‐regulated genes in an aryl hydrocarbon receptor nuclear translocator (Arnt)‐dependent manner
Authors: Choi Su Mi , Oh Hookeun , Park Hyunsung
Published: October 2008

We investigated hypoxia‐inducible factor (HIF)‐dependent changes in the expression of 5592 genes in response to hypoxia (0.1% O2, 16 h) by performing cDNA microarray analyses of mouse hepa1c1c7 and BpRc1 cells. BpRc1 cells are a hepa1c1c7 variant defective in HIF‐β/aryl hydrocarbon receptor nuclear translocator (Arnt), and are therefore unable to induce HIF target genes in response to hypoxia. By comparing hepa1c1c7 cells with BpRc1 cells, we were able to investigate hypoxia‐regulated gene expression as well as the role played by HIF in regulating the hypoxic‐dependent response of gene expression. This study identified 50 hypoxia‐induced genes and 36 hypoxia‐repressed genes. Quantitative PCR analysis of nine genes confirmed our ability to accurately analyze changes in hypoxia‐induced gene expression by microarray analysis. By comparing quantitative PCR analyses of these nine genes in BpRc1 and hepa1c1c7 cells, we determined that eight of the nine hypoxia‐induced genes are Arnt dependent. Additional quantitative PCR analyses of eight hypoxia‐repressed genes confirmed, with a 50% probability, that microarray analysis was able to predict hypoxia‐repressed gene expression. Only two of the four confirmed genes were found to be repressed in an Arnt‐dependent manner. Collectively, six of these 13 genes (46.2% probability) showed a pattern of expression consistent with the microarray analysis with regard to Arnt dependence. Finally, we investigated the HIF‐1α dependence of these 13 genes by quantitative PCR analysis in HIF‐1α knockdown 3T3‐L1 cells. These analyses identified novel hypoxia‐regulated genes and confirmed the role of Arnt and HIF‐1α in regulating their expression. These results identify additional HIF target genes and provide a more complete understanding of hypoxia signaling.
Identification of preferred substrate sequences for transglutaminase 1 – development of a novel peptide that can efficiently detect cross‐linking enzyme activity in the skin
Authors: Sugimura Yoshiaki , Hosono Masayo , Kitamura Miyako , Tsuda Tatsuya , Yamanishi Kiyofumi , Maki Masatoshi , Hitomi Kiyotaka
Published: October 2008

Transglutaminase 1 (TGase 1) is an essential enzyme for cornified envelope formation in stratified squamous epithelia. This enzyme catalyzes the cross‐linking of glutamine and lysine residues in structural proteins in differentiating keratinocytes. To gain insight into the preferred substrate structure of TGase 1, we used a phage‐displayed random peptide library to screen primary amino acid sequences that are preferentially selected by human TGase 1. The peptides selected as glutamine donor substrate exhibited a marked tendency in primary structure, conforming to the sequence: QxK/RψxxxWP (where x and ψ represent non‐conserved and hydrophobic amino acids, respectively). Using glutathione S‐transferase (GST) fusion proteins of the selected peptides, we identified several sequences as preferred substrates and confirmed that they were isozyme‐specific. We generated GST‐fused alanine mutants of the most reactive sequence (K5) to determine the residues that were critical for reactivity. Even in peptide form, K5 appeared to have high and specific reactivity as substrate. In situ analysis of mouse skin sections using fluorescence‐conjugated K5 peptide resulted in detection of TGase 1 activity with high sensitivity, but no signal was detected in a TGase 1‐null mouse. In conclusion, we were successful in generating a novel substrate peptide for sensitive detection of endogenous TGase 1 activity in the skin.
Chronic Administration of Ketamine Elicits Antidepressant‐Like Effects in Rats without Affecting Hippocampal Brain‐Derived Neurotrophic Factor Protein Levels
Authors: Garcia Lêda S. , Comim Clarissa M. , Valvassori Samira S. , Réus Gislaine Z. , Andreazza Ana Cristina , Stertz Laura , Fries Gabriel R. , Gavioli Elaine Cristina , Kapczinski Flavio , Quevedo João
Published: October 2008

Abstract:  A growing body of evidence has pointed to the blockade of the N‐methyl‐d‐aspartate (NMDA) receptor signaling as a potential therapeutic target for the treatment of major depression. The present study was aimed to evaluate behavioural and molecular effects of the chronic treatment with ketamine and imipramine in rats. To this aim, rats were 14 days treated once a day with ketamine (5, 10 and 15 mg/kg) and imipramine (10, 20 and 30 mg/kg) and then subjected to the forced swimming and open‐field tests. Ketamine and imipramine, at the all doses tested, reduced immobility time, and increased both climbing and swimming time of rats compared to the saline group, without affecting spontaneous locomotor activity. Brain‐derived neurotrophic factor (BDNF) hippocampal levels were assessed in imipramine‐ and ketamine‐treated rats by ELISA sandwich assay. Chronic administration of both drugs, ketamine and imipramine, did not modify BDNF protein levels in the rat hippocampus. In conclusion, our findings demonstrate for the first time that chronic administration of acute inactive doses of ketamine (5 mg/kg) becomes active after chronic treatment, while no signs of tolerance to the behavioural effects of ketamine were observed after chronic administration of acute active doses (10 and 15 mg/kg). Finally, these findings further support the hypothesis that NMDA receptor could be a new pharmacological target for the treatment of mood disorders.
Increased Hepatotoxicity and Cardiac Fibrosis in Cocaine‐Treated Butyrylcholinesterase Knockout Mice
Authors: Duysen Ellen G. , Li Bin , Carlson Michaela , Li Yi‐Fan , Wieseler Stacy , Hinrichs Steven H. , Lockridge Oksana
Published: October 2008

Abstract: In mice, cocaine is detoxified to inactive products by butyrylcholinesterase (BChE) and carboxylesterase. In human beings, cocaine detoxification is primarily by BChE. The focus of this investigation was to elucidate the importance of BChE in reducing pathophysiological effects following cocaine exposure. Previous studies examining the effects of cocaine on BChE deficient animals relied on chemical inhibition of BChE with tetraisopropyl pyrophosphoramide (iso‐OMPA). The creation of the BChE knockout mouse has provided a model for studying pathological effects of cocaine in mice free of chemical confounders. We hypothesized that mice with low or no BChE activity would have reduced cocaine metabolism, leading to hepatotoxicity and cardiomyopathy. A high‐resolutionin vivoimaging system recorded cardiac and respiratory function following treatment with a carboxylesterase inhibitor and a high dose of cocaine (100 mg/kg, intraperitoneally). The BChE−/− mice demonstrated depressed respiration through 12 hr after dosing and abnormal respiratory patterns consisting of a pause at full inspiration (apneusis), whereas BChE+/+ mice had recovered normal respiration rates by 30 min. after dosing and exhibited no apneusis. Liver and cardiac histology sections were analysed following a 20 mg/kg intraperitoneally dose of cocaine administered daily for 7 days. BChE–/– mice treated for 7 days with the chronic low dose showed significant hepatotoxicity and cardiac perivascular fibrosis compared to BChE+/+ mice. The observed functional changes following acute high‐dose and chronic low‐dose cocaine in BChE−/− and +/− mice warrants further investigation into the possibility of increased cocaine toxicity in human beings with BChE deficiency.
Antihyperlipidaemic Effect of Gymnema montanum: A Study on Lipid Profile and Fatty Acid Composition in Experimental Diabetes
Authors: Ramkumar Kunga Mohan , Vijayakumar Ramasamy Subramaniam , Ponmanickam Ponnirul , Velayuthaprabhu Shanmugan , Archunan Govindaraju , Rajaguru Palanisamy
Published: October 2008

Abstract:   In the present study, the antihyperlipidaemic efficacy of ethanol extract of Gymnema montanum leaves was investigated in alloxan‐induced diabetic rats and the effect was compared to standard hypoglycaemic drug, glibenclamide. Male adult albino Wistar rats were injected with freshly prepared solution of alloxan monohydrate (150 mg/kg body weight) to induce diabetes. After 2 weeks, the rats with moderate diabetes were administered G. montanum leaves (200 mg/kg body weight) for 21 days by gastric lavage, after which serum, liver and kidney samples were analysed for lipid profile, lipoprotein changes and fatty acid composition. While the alloxan‐induced diabetic rats showed a significant increase in the levels of cholesterol, triglycerides and free fatty acids, the levels in the animals treated with G. montanum leaves were considerably reduced and restored to near normal values. Antihyperlipidaemic effects of G. montanum leaves were found to be comparable with that of glibenclamide. Similarly, G. montanum leaves treatment resulted in reversal of alterations observed in the plasma lipoproteins (high‐density lipoprotein, low‐density lipoprotein and very high‐density lipoprotein‐cholesterol) and fatty acid composition in serum, liver and kidney of alloxan‐induced rats. Our study suggests that phytochemicals present in G. montanum may play an important role in suppressing the elevated lipid profile in diabetes and may be useful for the prevention and/or early treatment of diabetes‐associated hyperlipidaemia.