bluuebunny/crossref_metadata_2025_split huggingface dataset:
Thrips‐induced damage of chrysanthemum inflorescences: evidence for enhanced leakage of carotenoid pigments
Authors: Rhainds Marc , Doyon Josée , Rivoal Jean , Brodeur Jacques 
Published: March 2007

AbstractThe present study quantifies the leakage of carotenoid pigments in senescing inflorescences of chrysanthemum, Dendranthema grandiflora Tzelev (Asteraceae), infested or not with western flower thrips, Frankliniella occidentalis Pergande (Thysanoptera: Thripidae). The extractability of pigments was evaluated at repeated intervals by measuring the absorbance of immersion and crude extracts of inflorescences, using 447 nm as a diagnostic wavelength (absorption peak of lutein, a predominant carotenoid pigment in chrysanthemum with yellow inflorescences). The absorbance of immersion extracts increased over time and was higher for inflorescences infested with thrips than control inflorescences; the opposite trends were observed for crude extracts of inflorescences. These results suggest that both natural and thrips‐induced senescence of chrysanthemum inflorescences are characterized by enhanced extractability of pigments in organic solvents, most likely resulting from a disruption of cellular membranes. Thin layer chromatography analysis confirmed the presence of lutein in immersion extracts of thrips‐infested inflorescences, providing further support to the hypothesis that the mechanical damage caused by thrips is linked with a leakage of cellular content. An enhanced extractability of pigments in organic solvents may be a widespread phenomenon characterizing floral senescence in non‐climacteric plants.
Relationships between neurokinin receptor‐expressing interstitial cells of Cajal and tachykininergic nerves in the gut
Authors: Faussone‐Pellegrini Maria‐Simonetta 
Published: March 2007

AbstractThe so‐called interstitial cells of Cajal (ICC) are distributed throughout the muscle coat of the alimentary tract with characteristic intramual location and species‐variations in structure and staining. Several ICC sub‐types have been identified: ICC‐DMP, ICC‐MP, ICC‐IM, ICC‐SM. Gut motility is regulated by ICC and each sub‐type is responsible for the electrical activities typical of each gut region and/or muscle layer. The interstitial position of the ICC between nerve endings contain tachykinins. Three distinct tachykinin receptors (NK1r, NK2r and NK3r) have been demonstrated by molecular biology. Each of them binds with different affinities to a series of tachykinins (SP, NKA and NKB). In the ileum, SP‐immunoreactive (SP‐IR) nerve fibers form a rich plexus at the deep muscular plexus (DMP), distributed around SP‐negative cells, and ICC‐DMP intensely express the SP‐preffered receptor NK1r; conversely a faint NK1r‐IR is detected on the ICC‐MP and mainly after receptor internalization was induced by agonists. ICC‐IM are never stained in laboratory mammals, while those of the human, antrum are NK1r‐IR. RT‐PCR conducted on isolated idleal ICC‐MP and gastric ICC‐IM showed that these cells express NK1r and NK3r. Colonic ICC, except those in humans, do not express NK1r‐IR, at least in resting conditions. Outside the gut, NK1r‐IR cells were seen in the arterial wall and exocrine pancreas. In the mouse gut only, NK1r‐IR is present in non‐neuronal cells located within the intestinal villi, so‐called myoid cells, which are c‐kit‐negative and α‐smooth muscle actin‐positive. Immunohistochemistry and functional studies confirmed that ICC receive input from SP‐IR terminals, with differences between ICC sub‐types. In the rat, very early after birth, NK1r is expressed by the ICC‐DMP and SP by the related nerve varicosities. Studies on pathological conditions are few and those on mutant strains practically absent. It has only been reported that in the inflamed ileum of rats the NK1r‐IR ICC‐DMP disappear and that at the peak of inflammatory conditions ICC‐MP are NK1r‐IR. In ileum of mice with a mutation in the W locus, ICC‐DMP were seen to express c‐kit‐IR but not NK1‐IR, and SP‐IR innervation seems unchanged. In summary, there are distinct ICC populations, each of them under a different tachkininergic control and, likely, having different functions. Further studies are recommended at the aim of understanding ICC involvement in modulating/transmitting tachykininergic inputs.
Neuroglia in the inferior olivary nucleus during normal aging and Alzheimer's disease
Authors: Lasn H. , Winblad B. , Bogdanovic N. 
Published: March 2007

AbstractIt is likely that neuronal loss occurs in certain brain regions in Alzheimer's Disease (AD) without any neurofibrillary pathology. In the human principle inferior olivary nucleus (PO), we have shown that neuronal loss is about 34% (Lasn et al. Journal of Alzheimer Disease, 2001; 3: 159–168), but the fate of the neuroglial cells is unknown. Since the unique network of neurons and neuroglial cells and their cohabitation are essential for normal functioning of CNS, we designed a study to estimate the total number of oligodendrocytes and astrocytes in normally aged and AD brains. The study is based on 10 control and 11 AD post‐mortem human brains. An unbiased stereological fractionator method was used. We found significant oligodendroglial cell loss (46%) in AD as compared to control brains, while the total number of astrocytes showed a tendency to decrease. It is likely that the ratio of oligodendroglial cells to neurons remains unchanged even in degenerative states, indicating that oligodendroglial cells parallel neuronal loss. Astroglial cells did not increase in total number, but the ratio to neurons was significantly increased due to the neuronal loss. Using a novel unbiased quantitative method, we were able to describe significant oligodendroglial loss in the PO but the pathogenic mechanism behind remains unknown.
Caveolin‐3 inhibits growth signal in cardiac myoblasts in a Ca2+‐dependent manner
Authors: Fujita Takayuki , Otsu Kouji , Oshikawa Jin , Hori Hideaki , Kitamura Hitoshi , Ito Takaaki , Umemura Satoshi , Minamisawa Susumu , Ishikawa Yoshihiro 
Published: March 2007

AbstractCaveolin, a major protein component of caveolae, directly interacts with multiple signaling molecules, such as Ras and growth factor receptors, and inhibits their function. However, the role of the second messenger system in mediating this inhibition by caveolin remains poorly understood. We examined the role of Ca2+ ‐dependent signal in caveloin‐mediated growth inhibition using a rat cardiac myoblast cell line (H9C2), in which the expression of caveolin‐3, the muscle specific subtype, can be induced using the LacSwitch system. Upon induction with IPTG and serum‐starvation, the expression of caveolin‐3 was increased by 3.3‐fold relative to that of mock‐induced cells. The recombinant caveolin‐3 was localized to the same subcellular fraction as endogenous caveolin‐3 after sucrose gradient purification. Angiotensin II enhanced ERK phosphorylation, but this enhancement was significantly decreased in caveolin‐3‐induced cells in comparison to that in mock‐induced cells. Similarly, when cells were stimulated with fetal calf serum, DNA synthesis, as determined by [3H]‐thymidine incorporation, was significantly decreased in caveolin‐3‐induced cells. When cells were treated with Ca2+ chelator (BAPTA and EGTA), however, this attenuation was blunted. Calphostin (PKC inhibitor), but not cyclosporine A treatment (calcineurin inhibitor), blunted this attenuation in caveolin‐3 induced cells. Our findings suggest that caveolin exhibits growth inhibition in a Ca2+‐dependent manner, most likely through PKC, in cardiac myoblasts.
Interstitial Cajal‐like cells (ICLC) in atrial myocardium: ultrastructural and immunohistochemical characterization
Authors: Hinescu M. E. , Gherghiceanu Mihaela , Mandache E. , Ciontea Sanda M. , Popescu L. M. 
Published: March 2007

AbstractWe have previously reported (Hinescu & Popescu, 2005) the existence of interstitial Cajal‐like cells (ICLC), by transmission electron microscopy, in human atrial myocardium. In the present study, ICLC were identified with non‐conventional light microscopy (NCLM) on semi‐thin sections stained with toluidine blue and immunohistochemistry (IHC) for CD117/c‐kit, CD34, vimentin and other additional antigens for differential diagnosis. Quantitatively, on semi‐thin sections, ICLC represent about 1–1.5% of the atrial myocardial volume (vs.±45% working myocytes, ˜2% endothelial cells, 3–4% for other interstitial cells, and the remaining percentage: extracellular matrix). Roughly, there is one ICLC for 8–10 working atrial myocytes in the intercellular space, beneath the epicardium, with a characteristic (pyriform, spindle or triangular) shape. These ICLC usually have 2–3 definitory processes, emerging from cell body, which usually embrace atrial myocytes (260 nm average distance plasmalemma/sarcolemma) or establish close contact with nerve fibers or capillaries (˜420 nm average distance to endothelial cells). Cell prolongations are characteristic: very thin (mean thickness = 0.150±0.1 μm), very long for a non‐nervous cell (several tens of μm) and moniliform (uneven caliber). Stromal synapses between ICLC and other interstitial cells (macrophages) were found (e.g. in a multicontact type synapse, the average synaptic cleft was ˜65 nm). Naturally, the usual cell organelles (mitochondria, smooth and rough endoplasmic reticulum, intermediate filaments) are relatively well developed. Caveolae were also visible on cell prolongations. No thick filaments were detected. IHC showed that ICLC were slightly and inconsistently positive for CD117/c‐kit, variously co‐expressed CD34 and EGF receptor, but appeared strongly positive for vimentin, along their prolongations. Some ICLC seemed positive for α‐smooth muscle actin and tau protein, but were negative for nestin, desmin, CD13 and S‐100.In conclusion, we provide further evidence of the existence of ICLC in human atrial myocardium, supporting the possible ICLC role in pacemaking, secretion (juxta‐ and/or paracrine), intercellular signaling (neurons and myocytes). For pathology, ICLC might as well be ‘players’ in arrhythmogenesis and atrial remodeling.
Blocking translocation of cell surface molecules from the ER to the cell surface by intracellular antibodies targeted to the ER
Authors: Böldicke Thomas 
Published: March 2007

Introduction
Generation of ER‐targeted intrabodies
Blocking translocation of growth factor receptors
ER intrabodies against infectious diseases
Abrogation of MHC I molecules
ER intrabodies targeting integrins
ER intrabodies targeting secretory proteins involved in Alzheimer's and Prion disease
Intrabody gene delivery
Conclusions

AbstractIntracellular antibodies (intrabodies) constitute a potent tool to neutralize the function of target proteins inside specific cell compartments (cytosol, nucleus, mitochondria and ER). The intrabody technology is an attractive alternative to the generation of gene‐targeted knockout animals and complements or replaces knockdown techniques such as antisense‐RNA, RNAi and RNA aptamers. This article focuses on intrabodies targeted to the ER. Intracellular anti‐bodies expressed and retained inside the ER (ER intrabodies) are shown to be highly efficient in blocking the translocation of secreted and cell surface molecules from the ER to the cell surface.The advantage of ER intrabodies over cytoplasmic intrabodies is that they are correctly folded and easier to select. A particular advantage of the intrabody technology over existing ones is the possibility of inhibiting selectively post‐translational modifications of proteins.The main applications of ER intrabodies so far have been (i) inactivation of oncogenic receptors and (ii) functional inhibition of virus envelope proteins and virus‐receptor molecules on the surface of host cells.In cancer research, the number of in vivo mouse models for evaluation of the therapeutic potential of intrabodies is increasing.In the future, endosomal localized receptors involved in bacterial and viral infections, intracellular oncogenic receptors and enzymes involved in glycosylation of tumour antigens might be new targets for ER intrabodies.
Correlation between cardiac oxidative stress and myocardial pathology due to acute and chronic norepinephrine administration in rats
Authors: Neri Margherita , Cerretani Daniela , Fiaschi Anna Ida , Laghi Pasini Franco , Lazzerini Pietro Euea , Maffione Angela Bruna , Micheli Lucia , Bruni Giancarlo , Nencini Cristina , Giorgi Giorgio , D'Errico Stefano , Fiore Carmela , Pomara Cristoforo , Riezzo Irene , Turillazzi Emanuela , Fineschi Vittorio 
Published: March 2007

Abstract Background: To investigate the cardiotoxic role of reactive oxygen species (ROS) and of products derived from catecholamines auto‐oxidation, we studied: (1) the response of antioxidant cardiac cellular defence systems to oxidative stress induced by norepinephrine (NE) administration, (2) the effect of NE administration on cardiac β1‐adrenergic receptors by means of receptor binding assay, (3) the cellular morphological alterations related to the biologically cross‐talk between the NE administration and cytokines [tumor necrosis factor‐alpha (TNF‐α), monocyte chemotactic protein‐1 (MCP‐1), interleukins IL6, IL8, IL10]Methods and Results: A total of 195 male rats was used in the experiment. All animals underwent electrocardiogram (EKG) before being sacrificed. The results obtained show that NE administration influences the antioxidant cellular defence system significantly increasing glutathione peroxidase (GPx) activity, glutathione reductase (GR) and superoxide dismutase (SOD). The oxidized glutathione (GSH/GSSG) ratio significantly decreases and malondialdehyde (MDA) levels increase showing a state of lipoperoxidation of cardiac tissue. We describe a significant apoptotic process randomly sparse in the damaged myocardium and the effect of ROS on the NE‐mediated TNF‐α, MCP‐1, and IL6, IL8, IL10 production. Conclusions: Our results support the hypothesis that catecholamines may induce oxidative damage through reactive intermediates resulting from their auto‐oxidation, irrespective of their interaction with adrenergic receptors, thus representing an important factor in the pathogenesis of catecholamines‐induced cardiotoxicity. The rise of the cardioinhibitory cytokines may be interpreted as the adaptive response of jeopardized myocardium with respect to the cardiac dysfunction resulting from NE injection.
Autonomously vascularized cellular constructs in tissue engineering: opening a new perspective for biomedical science
Authors: Polykandriotis E. , Arkudas A. , Horch R.E. , Stürzl M. , Kneser U. 
Published: March 2007

Introduction
Cellular homeostasis and vascularization
Importance of vascularization: historical review
Multicellular implants in vivo
Models of axial vascular induction
Development of the AV loop model
Angiogenesis in multicellular implants in vivo: authors' experience
Physiology of angiogenesis in the AV loop
Axially vascularized constructs as tools for research
Conclusion

AbstractIn tissue engineering cell cultures play a crucial role besides the matrix materials for the end of substituting lost tissue functions. The cell itself is situated at the cross‐roads leading to different orders of scale, from molecule to organism and different levels of function, from biochemistry to macrophysiology. Extensive in vitro investigations have dissected a vast amount of cellular phenomena and the role of a number of bioactive substances has been elucidated in the past. Further, recombinant DNA technologies allow modulation of the expression profiles of virtually all kinds of cells. However, issues of vascularization in vivo limit transferability of these observations and restrict upscaling into clinical applications. Novel in vivo models of vascularization have evolved inspired from reconstructive microsurgical concepts and they encompass axial neovascularization by means of vascular induction. This work represents a brief description of latest developments and potential applications of neovascularization and angiogenesis in tissue engineering.
PIKE GTPase are phosphoinositide‐3‐kinase enhancers, suppressing programmed cell deathPIKE GTPase are phosphoinositide‐3‐kinase enhancers, suppressing programmed cell death
Authors: Chan Chi Bun , Ye Keqiang , Chan Chi Bun , Ye Keqiang 
Published: March 2007

Introduction
The structure, tissue distribution and cellular localization of PIKEs
Mitogenic PIKE‐S signaling in the nucleus
Anti‐apoptotic function of PIKE‐L in neurons
Role of PIKE‐L in merlin inhibited growth suppression
Phosphoinositol lipids as a feedback regulator to PIKE‐L activation and translocation
Anti‐apoptotic activity of PIKE‐A in cancers
PIKE‐A as the physiological substrate of Fyn
Perspective remarks

AbstractPhosphoinositide‐3‐kinase enhancers (PIKE) are GTP‐binding proteins that posses anti‐apoptotic functions. The PIKE family includes three members, PIKE‐L, PIKE‐S and PIKE‐A, which are originated from a single gene (CENTG1) through alternative splicing or differential transcription initiation. Both PIKE‐S and PIKE‐L bind to phosphoinositide‐3‐kinase (PI3K) and enhance its activity. PIKE‐A does not interplay with PI3K. Instead, it interacts with the downstream effector Akt and promotes its activity. These actions are mediated by their GTPase activity. Because both PI3K and Akt are important effectors in the growth factor‐mediated signaling which triggers cellular growth and acts against apoptosis, PIKEs therefore serve as the molecular switch that their activation are crucial for growth factors to exert their physiological functions. In this review, the current understanding of different PIKE isoforms in growth factors‐induced anti‐apoptotic function will be discussed. Moreover, the role of PIKE in the survival and invasion activity of cancer cells will also be introduced.
Reversible transfection of human melanocytes mediated by Cre/loxP site‐specific recombination system and SV40 large T antigen
Authors: Ying Wang , Fei Hao , Jun Deng , Xi‐chuan Yang , Bai‐yu Zhong , Qing‐yi Ye 
Published: March 2007

Abstract:  Objective:  To study the reversible transfection of human melanocytes mediated by simian virus 40 large T antigen (SV40LTAg) and Cre/loxP site‐specific recombination system.Methods:  The reconstructed SV40LTAg‐EGFP‐neo‐loxP vector was transfected into primary cultured human melanocytes with SofastTM transfection reagent and the positive cells were selected using G418. After expanding culture of these positive cell clones, the expression of SV40LTAg was detected by polymerase chain reaction (PCR), reverse transcription‐polymerase chain reaction (RT‐PCR) and immunofluorescent method. After that, these positive cells were infected by virus supernatant of Cre‐ERT2 retrovirus vector and Cre recombinase was induced to act by tamoxifen. On the 6th and 10th day after Cre recombinase acting, the expression of SV40LTAg was detected using the same methods as above, and cell tumorigenicity was studied using soft agar assay, athymic mouse study and karyotype analysis. On 10th day after tamoxifen treatment, cell biological characters were identified with immunofluorescent staining and transmission electron microscopy. Then these cells were transplanted into vitiligo animal model to observe their melanogenesis ability in vivo.Results:  The genome DNA and total RNA were isolated from the positive cells transfected by SV40LTAg (designated as MCT) and specific 288 bp fragment was amplificated using PCR and RT‐PCR methods. The results of immunofluorescence confirmed the expression of SV40LTAg in cell nucleus. On the 6th day after tamoxifen treatment in infected cells by Cre‐ERT2 retrovirus vector (designated as MCT‐Cre), there could be detected SV40LTAg expression, but on 10th day, there could not be detected SV40LTAg expression in cells. These results showed that the excised efficiency of Cre recombinase increased along with time prolongation, and would obtain complete recombination efficiency. The identification of MCT‐Cre cell biological characters showed that these cells had normal parent‐cell‐like cell phenotype and no tumorigenicity in vitro. The pigmentation started in 4 weeks and formed black macula in 3 months after grafting. The pathological results showed that there had been significant melanocytes and melanin accumulation in epidermis and some hair follicle in transplanted area, which confirmed that MCT‐Cre had melanogenesis function in vivo.Conclusion:  Human melanocytes could be mediated by reversible transfection by SV40LTAg and Cre/loxP site‐specific recombination system, which had stable parent‐cell‐like phenotypic characters and no tumorigenicity in vitro; moreover, these cells still had melanogenesis function in vivo.
Immunological responses and viral modulatory effects of vaccination with recombinant modified vaccinia virus Ankara (rMVA) expressing structural and regulatory transgenes of simian immunodeficiency virus (SIVmac32H/J5M)
Authors: Berry N. , Stebbings R. , Brown S. , Christian P. , Thorstensson R. , Ahmed R.K. , Davis L. , Ferguson D. , D'Arcy N. , Elsley W. , Hull R. , Lines J. , Wade‐Evans A. , Stott J. , Almond N. 
Published: March 2007

AbstractBackground  The immunogenicity and protective efficacy of recombinant modified vaccinia virus Ankara (rMVA) vectors expressing structural (gag/pol, env) and regulatory (tat, rev, nef) genes of SIVmac251/32H‐J5 (rMVA‐J5) were assessed.Methods  Immunization with rMVA constructs (2.5 × 107 IU) 32, 20 and 8 weeks pre‐challenge was compared with 32 and 20 weeks but with a final boost 8 weeks pre‐challenge with 2 × 106 fixed‐inactivated HSC‐F4 cells infected with SIVmac32H. Controls received rMVA vectors expressing an irrelevant transgene or were naïve challenge controls. All received 10 MID50 SIVmac32H/J5 intravenously.Results  Vaccinates immunized with rMVA‐J5 exhibited significant, albeit transient, control of peak primary viraemia despite inconsistent and variable immune responses elicted by vaccination. Humoral and cellular responses to Env were most consistent, with lower responses to Nef, Rev and Tat. Increasing titres of anti‐vaccinia neutralizing antibodies reflected the number and dose of rMVA inoculations.Conclusions  Improved combinations of viral vectors are required to elicit appropriate immune responses to control viral replication.
Induction of Apoptosis by Ginger in HEp‐2 Cell Line Is Mediated by Reactive Oxygen Species
Authors: Vijaya Padma Viswanadha , Arul Diana Christie Swamidurai , Ramkuma Kunga Mohan 
Published: March 2007

Abstract:   Ginger (Zingiber officinale Roscoe, Zingiberaceae) is a commonly used medicinal herb throughout the world. Although some studies have demonstrated its antitumour activities on cancer cells in vitro and in vivo, the exact mechanism is not fully elucidated. Hence, the present study was designed to examine the in vitro cytotoxic activities of saline extract prepared from ginger extract on HEp‐2 cell line. The cytotoxic effect of the drug was confirmed by 3‐(4,5‐dimethyl thiazol‐2‐yl)‐2,5‐diphenyl tetrazolium bromide assay and cell counting and estimation of protein, DNA and RNA. Meanwhile, propidium iodide staining and agarose gel electrophoresis were performed for determining the induction of apoptosis. In addition, superoxide radical generation, nitrite formation and glutathione studies show involvement of free radicals. The present results show that the extract exerts dose‐dependent suppression of cell proliferation; the IC50 value was found to be 900 µg/ml. At a dose of 250 µg/ml, marked morphological changes including cell shrinkage and condensation of chromosomes were observed. Agarose gel electrophoresis of DNA from HEp‐2 cells treated with 250 µg/ml ginger powder for 24 hr showed marked DNA ladder pattern. The involvement of free radicals was confirmed by increased superoxide production, decreased nitrate formation and depletion of glutathione in ginger‐treated cells. Further screening of active components using gas chromatography–mass spectrometry analyses showed the presence of clavatol, geraniol and pinostrobin in the extract. The results of the present study suggest that ginger might be useful as a potential antitumour agent.