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Selected AbstractsScreening of urocanic acid isomers in human basal and squamous cell carcinoma tumors compared with tumor periphery and healthy skinEXPERIMENTAL DERMATOLOGY, Issue 10 2008Juan Manuel Decara Abstract:,Trans -urocanic acid is a major chromophore for ultraviolet (UV) radiation in human epidermis. The UV induces photoisomerization of trans -urocanic acid (tUCA) form to cis -urocanic acid (cUCA) and has been reported as an important mediator in the immunosuppression induced by UV. This immunomodulation has been recognized as an important factor related to skin cancer development. This is the first time that UCA isomers have been measured in epidermis of skin biopsies from patients with squamous cell carcinoma (SCC) and with basal cell carcinoma (BCC) and compared with the tumor periphery and biopsies of healthy photoexposed and non-photoexposed skin as controls. The UCA isomers were separated and quantified by high performance liquid chromatography. Analysis of UCA in healthy skin showed significant increase in total UCA content in non-photoexposed body sites compared with highly exposed skins. In contrast, the percentage of cUCA was higher in photoexposed body sites. Maximal levels of cUCA were found in cheek, forehead and forearm and lower levels in abdomen and thigh. No differences were found in total UCA concentration between the tumor samples and healthy photoexposed skin. However, differences were found in relation between isomers. Higher levels of cUCA were detected in SCC biopsies (44% of total UCA) compared with samples of BCC and that of healthy photoexposed skin (30%). These results suggest that the UV radiation exposure, a main factor in development of SCC can be mediated, apart from direct effect to cells (DNA damage), by immunosuppression pathways mediated by high production of cUCA. [source] Fractal analysis of Daphnia motion for acute toxicity bioassayENVIRONMENTAL TOXICOLOGY, Issue 5 2002Nobuaki Shimizu Abstract To quantify individual behavioral responses to toxic chemicals, the swimming motion of individual Daphniamagna was continuously monitored using a motion analysis system. The fractal dimension was introduced to compare the straightness or complexity of the swimming trajectory before and after exposure to toxic chemicals. Analysis indicated that the swimming trajectory of individual Daphnia has a fractal structure. The basal fractal dimension in the control medium was 1.35±0.01 (n = 50 Daphnia). Exposure to CuSO4 (10 ,g/L), organophosphorus (Dichlorvos; 10 ,g/L), and carbamate (Propoxur; 500 ,g/L) pesticide caused a significant increase in the fractal dimension with a latency of 60 min, reaching a maximal level of 2.26±0.34, 2.43±0.19, and 2.51±0.21, respectively, after a 120-min exposure. The magnitude of the change in the fractal dimension was related to the toxic chemical concentration and the exposure time. Threshold concentrations determined at 60 min of exposure were 10 ,g/L for CuSO4, 5 ,g/L for Dichlorvos, and 500 ,g/L for Propoxur. The toxicity index (EC50) values after 120 min of exposure were 6.31 ,g/L, 7.64 ,g/L, and 466 ,g/L for CuSO4, Dichlorvos, and Propoxur, respectively. Thus, the fractal dimension seems useful for analyzing and comparing complex trails, such as swimming trajectories, which could be used as the endpoint for an acute bioassay. © 2002 Wiley Periodicals, Inc. Environ Toxicol 17: 441,448, 2002; Published online in Wiley InterScience (www.interscience.wiley.com). DOI 10.1002/tox.10077 [source] Productive Chlamydia trachomatis lymphogranuloma venereum 434 infection in cells with augmented or inactivated autophagic activitiesFEMS MICROBIOLOGY LETTERS, Issue 2 2009Niseema Pachikara Abstract Autophagy, a eukaryotic cellular activity leading to the degradation of cellular components, serves as a defense mechanism against facultative intracellular bacteria as well as a growth niche for the obligate intracellular bacterium Coxiella burnetii. We here demonstrate that the obligate intracellular bacterial pathogen Chlamydia trachomatis lymphogranuloma venereum strongly induced autophagy in the middle of the chlamydial developmental cycle (24 h after infection), a time point with maximal level of chlamydial replication, but not during the early stages with low overall chlamydial metabolism (before 8 h). No autophagy induction was evident in cells exposed to heat- and UV-inactivated elementary bodies (EBs, the infectious form of Chlamydia) or to inocula from which EBs had been removed before inoculation. Blocking chlamydial development with chloramphenicol also prevented autophagy induction in cells infected with infectious EBs. It appears that autophagy is activated primarily in response to the metabolic stress consequent to chlamydial replication. However, autophagy-defective ATG5,/, cells supported chlamydial development as efficiently as autophagy-proficient ATG5+/+ cells. [source] In the hypoxic central nervous system, endothelial cell proliferation is followed by astrocyte activation, proliferation, and increased expression of the ,6,4 integrin and dystroglycanGLIA, Issue 10 2010Longxuan Li Abstract Cerebral hypoxia induces a profound angiogenic response in the central nervous system (CNS). Using a mouse model of chronic cerebral hypoxia, we previously demonstrated that angiogenic vessels in the hypoxic CNS show marked upregulation of the extracellular matrix (ECM) protein fibronectin, along with increased expression of its major receptor, ,5,1 integrin on brain endothelial cells (BEC). As cerebral hypoxia also leads to glial activation, the aim of the current study was to define the temporal relationship between BEC responses and glial cell activation in this model of cerebral hypoxia. This revealed that BEC fibronectin/,5,1 integrin expression and proliferation both reached maximal level after 4-day hypoxia. Interestingly, up to 4-day hypoxia, all dividing cells were BEC, but at later time-points proliferating astrocytes were also observed. GFAP staining revealed that hypoxia induced marked astrocyte activation that reached maximal level between 7- and 14-day hypoxia. As newly formed cerebral capillaries require ensheathment by astrocyte end-feet to acquire mature brain endothelium characteristics, we next examined how expression of astrocyte end-feet adhesion molecules is regulated by hypoxia. This showed that the astrocyte adhesion receptors ,6,4 integrin and dystroglycan were both markedly upregulated, with a time-course that closely resembled astrocyte activation. Taken together, this evidence shows that cerebral hypoxia promotes first an endothelial response, in which fibronectin promotes BEC proliferation. This is then followed by an astrocyte response, involving astrocyte activation, proliferation, and reorganization of astrocyte end-feet, which correlates with increased expression of astrocyte end-feet adhesion molecules. © 2010 Wiley-Liss, Inc. [source] The contribution of stone cover to biological activity in the Negev desert, IsraelLAND DEGRADATION AND DEVELOPMENT, Issue 1 2001I. Lahav (Lavian) Abstract Ancient valley agriculture in the northern Negev highlands was based on the principle of directed collection of water and eroded material from the slopes and their consequent flow towards the valleys. The stones on these slopes were therefore removed and/or collected into piles known as ,grape mounds'. The aim of this study was to understand the contribution of stone cover and slope-facing to biological activity in soil. Soil samples from a depth of 0,5,mm from the soil surface were collected during the study period (December 1994,March 1996) from northern and southern hill slopes, from under limestones and between stones. Soil moisture, organic matter, chlorophyll-a and soil respiration were determined. The results obtained in field and laboratory studies demonstrated differences between the northern and southern slopes. The stone cover on the northern slope made up 33 per cent and in the southern slope 23 per cent, stone size ranging from 15,50,cm2 and 15,35,cm2, respectively. Soil moisture content varied from 12 per cent in December 1994 on both slopes to one-quarter of the initial value during the dry period. Organic matter content reached a maximal level of 14 per cent and 16 per cent on the northern and southern slopes, respectively. Values of chlorophyll-a on both the northern and southern slopes were 0.38,,g,g,1 dry soil during the wet season, decreasing to 0.05,,g,g,1 dry soil during the dry period. Soil samples from under the stones on both slopes produced high levels of CO2, ranging between 50 and 100,,g CO2,g;,1 dry soil h,1, whereas in the control samples the levels ranged between 30 and 70,,g CO2,g,1 dry soil h,1. In conclusion, the stone cover apparently plays an important role in the maintenance of biological activity through its contribution to slope biotope stability. Copyright © 2001 John Wiley & Sons, Ltd. [source] Immunocytochemical analysis of the circadian clock protein in mouse hepatocytesMICROSCOPY RESEARCH AND TECHNIQUE, Issue 5 2003Manuela Malatesta Abstract Many biochemical, physiological, and behavioral processes in organisms ranging from prokaryotes to humans exhibit circadian rhythms, defined as cyclic oscillations of about 24 hours. The mechanism of the cellular circadian clock relies on interlocking positive and negative transcriptional/translational feedback loops based on the regulated expression of several genes. Clock is one of these genes and its transcript, CLOCK protein, is a transcription factor belonging to the bHLH-PAS family. In mammals the clock gene is expressed in several tissues, including the liver. In the present study, we analyzed by means of quali-quantitative immunoelectron microscopy the fine intracellular distribution of the CLOCK protein in mouse hepatocytes during the daily cycle. We demonstrated that CLOCK protein is mostly located in the cell nucleus, where it accumulates on perichromatin fibrils, representing the in situ form of nascent pre-mRNA, while condensed chromatin and nucleoli contain lower amounts of protein. Moreover, we found that CLOCK protein shows circadian oscillations in these nuclear compartments, peaking in late afternoon. At this time the hepatic transcriptional rate reaches the maximal level, thus suggesting an important role of CLOCK protein in the regulation of liver gene expression. Microsc. Res. Tech. 61:414,418, 2003. © 2003 Wiley-Liss, Inc. [source] Temporal patterns of genetic variation across a 9-year-old aerial seed bank of the shrub Banksia hookeriana (Proteaceae)MOLECULAR ECOLOGY, Issue 13 2005LUKE G. BARRETT Abstract The pattern of accumulation of genetic variation over time in seed banks is poorly understood. We examined the genetic structure of the aerial seed bank of Banksia hookeriana within a single 15-year-old population in fire-prone southwestern Australia, and compared genetic variation between adults and each year of a 9-year-old seed bank using amplified fragment length polymorphism (AFLP). B. hookeriana is well suited to the study of seed bank dynamics due to the canopy storage of its seeds, and because each annual crop can be identified. A total of 304 seeds from nine crop years and five maternal plants were genotyped, along with 113 plants from the adult population. Genetic variation, as assessed by the proportion of polymorphic markers (Pp) and Shannon's index (I), increased slightly within the seed bank over time, while gene diversity (Hj), did not change. Pp, I, and Hj all indicated that genetic variation within the seed bank quickly approached the maximal level detected. Analysis of molecular variance revealed that less than 4% of variation could be accounted for by variation among seeds produced in different years, whereas there was greater differentiation among maternal plants (12.7%), and among individual seeds produced by different maternal plants (83.4%). With increasing population age, offspring generated each year were slightly more outbred, as indicated by an increase in the mean number of nonmaternal markers per offspring. There were no significant differences for Hj or I between adults and the seed bank. Viability of seeds decreased with age, such that the viability of 9-year-old seeds was half that of 2-year-old seeds. These results suggest that variable fire frequencies have only limited potential to influence the amount of genetic variation stored within the seed bank of B. hookeriana. [source] Conservative treatment of L -asparaginase-associated lipid abnormalities in children with acute lymphoblastic leukemia,PEDIATRIC BLOOD & CANCER, Issue 5 2010Hofit Cohen MD Abstract Objective To determine the incidence and clinical consequences of asparaginase-associated lipid abnormalities in children with acute lymphoblastic leukemia (ALL). Methods Sixty-five newly diagnosed children and adolescents aged 0.4,21 years with ALL or lymphoblastic lymphoma were retrospectively evaluated for lipid abnormalities. They were treated according to the ALLIC-BFM 2002 protocol between 2002 and 2005. Fasting cholesterol levels were measured in all patients and triglycerides (TG) in 42/65 patients. Results Prior to treatment, mean cholesterol level was 149,±,50,mg/dl, and increased to maximal level 274,±,124,mg/dl during treatment. Mean TG level during treatment was 459,±,526,mg/dl (range 54,3,009). Twelve patients (28%) had TG levels <200,mg/dl, 18 (43%) had 200,400,mg/dl, 3 (7%) had 400,600,mg/dl, 4 (10%) between 600 and 1,000,mg/dl, and 5 (12%) patients had >1,000,mg/dl. No association was found between TG levels and age or gender. One of the 12 patients with TG >400,mg/dl developed left saggital sinus thrombosis and left frontal lobe infarct. TG level at the time of the event was 2,640,mg/dl. None of the five patients with TG levels >1,000,mg/dl developed pancreatitis. Children with TG levels between 400 and 600,mg/dl were treated by fasting. Fibrates and heparin were added to those with levels >600,mg/dl. Lipid abnormalities normalized in all children upon completion of asparaginase treatment. Conclusions Abnormalities of lipid profile in children with ALL during asparaginase therapy are relatively common. We recommend measuring TG before and during asparaginase treatment. Initiation of conservative treatment could prevent further increase of TG and decrease the risk of potential complications. Pediatr Blood Cancer 2010;54:703,706. © 2010 Wiley-Liss, Inc. [source] Xanthopterin in the Oriental Hornet (Vespa orientalis): Light Absorbance Is Increased with Maturation of Yellow Pigment GranulesPHOTOCHEMISTRY & PHOTOBIOLOGY, Issue 4 2009Marian Plotkin The Oriental hornet bears both brown and yellow colors on its cuticle. The brown component is contributed by the pigment melanin, which is dispersed in the brown cuticle and provides protection against insolation, while the yellow-colored part contains within pockets in the cuticle granules possessing a yellow pigment. These yellow granules (YG) are formed about 2 days prior to eclosion of the imago, and their production continues for about 3 days posteclosion. Xanthopterin is the main component of the granule and lends it its yellow color. Xanthopterin produces a characteristic excitation/emission maximum at 386/456 nm. Characterization by use of mass spectrometry showed the compound to have a molecular ion of 179, as expected from xanthopterin. Spectroscopic examination of the absorption of an entire stripe of yellow cuticle in the course of its metamorphosis revealed that the absorption steadily increases throughout the process to a maximal level of absorption about 3 days posteclosion. In the absence of the YG, the cuticle is permeable to the passage of all wavelengths within the visible range and to the UV range (290,750 nm) in all age groups of hornets. The newly ecloded hornets depart the nest to engage in activities requiring exposure to insolation only as the process of granule formation terminates, namely, when the layer of YG in the cuticle suffices to absorb all the harmful UV radiation. [source] Changes in mesophyll anatomy and sink,source relationships during leaf development in Quercus glauca, an evergreen tree showing delayed leaf greeningPLANT CELL & ENVIRONMENT, Issue 5 2003S.-I. MIYAZAWA ABSTRACT Changes in mesophyll anatomy, gas exchange, and the amounts of nitrogen and cell wall constituents including cellulose, hemicellulose and lignin during leaf development were studied in an evergreen broad-leaved tree, Quercus glauca, and in an annual herb, Phaseolus vulgaris. The number of chloroplasts per whole leaf in P. vulgaris increased and attained the maximal level around 10 d before full leaf area expansion (FLE), whereas it continued to increase even after FLE in Q. glauca. The increase in the number of palisade tissue cells per whole leaf continued until a few days before FLE in Q. glauca, but it had almost ceased by 10 d before FLE in P. vulgaris. The radius and height of palisade tissue cells in Q. glauca, attained their maximal levels at around FLE whereas the thickness of the mesophyll cell wall and concentrations of the cell wall constituents increased markedly after FLE. These results clearly indicated that, in Q. glauca, chloroplast development proceeded in parallel with the cell wall thickening well after completion of the mesophyll cell division and cell enlargement. The sink,source transition, defined to be the time when the increase in daily carbon exchange rate exceeds the daily increase in leaf carbon content, occurred before FLE in P. vulgaris but after FLE in Q. glauca. During leaf area expansion, the maximum daily increase in nitrogen content on a whole leaf basis (the maximum leaf areas were corrected to be identical for these species) in Q. glauca was similar to that in P. vulgaris. In Q. glauca, however, more than 70% of nitrogen in the mature leaf was invested during its sink phase, whereas in P. vulgaris it was 50%. These results suggest that Q. glauca invests nitrogen for cell division for a considerable period and for chloroplast development during the later stages. We conclude that the competition for nitrogen between cell division and chloroplast development in the area of expanding leaves can explain different greening patterns among plant species. [source] The responsive expression of heat shock protein 22 gene in zhikong scallop Chlamys farreri against a bacterial challengeAQUACULTURE RESEARCH, Issue 2 2010Lei Zhang Abstract HSP22 is a member of a small HSP subfamily contributing to the growth, transformation and apoptosis of the cell as well as acting as a molecular chaperone. In the present study, CfHSP22 cDNA was cloned from Chlamys farreri by the rapid amplification of cDNA ends technique. The full-length cDNA of CfHSP22 was of 1279 bp, consisting of a 5,-terminal untranslated region (5,UTR) of 122 bp, a 3,UTR of 581 bp with a canonical polyadenylation signal sequence AATAAA and a poly(A) tail, and an open reading frame of 576 bp encoding a polypeptide with a molecular mass of 22.21 kDa and a predicted isoelectric point of 9.69. There was an ,-crystallin domain, a hallmark of the sHSP subfamily, in the C-terminus, and the deduced amino acid sequence of CfHSP22 showed high similarity to previously identified HSP22s. CfHSP22 was constitutively expressed in the haemocyte, muscle, kidney, gonad, gill, heart and hepatopancreas, and the expression level in the hepatopancreas was higher than that in the other tissues. CfHSP22 transcription was up-regulated and reached a maximal level at 12 h after the bacterial challenge, and then declined progressively to the original level at 48 h. These results suggested that CfHSP22 perhaps play a critical role in response to the bacterial challenge in haemocytes of scallop C. farreri. [source] Expression and function of the ST2 gene in a murine model of allergic airway inflammationCLINICAL & EXPERIMENTAL ALLERGY, Issue 10 2002K. Oshikawa Summary Background We have recently reported that soluble ST2 protein levels are elevated in the sera of patients with asthma, and correlate well with the severity of asthma exacerbation. However, the role, function, and kinetics of soluble ST2 expression in asthma remain unclear. Objective The objective of the present study was to clarify the function and kinetics of soluble murine (m) ST2 expression in a murine asthma model. Methods We analyzed the kinetics of gene and protein expression of mST2 in sera or lung tissue after allergen (ovalbumin; OVA) challenge in a murine model of allergic airway inflammation, the effects of mST2 protein on OVA-induced Th2 cytokine production in vitro from splenocytes of sensitized mice, and the effects of soluble mST2 on Th2-dependent allergic airway inflammation by in vivo gene transfer of mST2. Results Serum mST2 protein levels increased to the maximal level 3 h after the allergen challenge, before serum IL-5 levels peaked. The mRNA expression of mST2 in lung tissue was induced after the allergen challenge, while that in the spleen was constitutively detected. Furthermore, pre-treatment with mST2 protein significantly inhibited the production of IL-4 and IL-5, but not IFN-,, from OVA-stimulated splenocytes in vitro, and intravenous mST2 gene transfer resulted in a drastic reduction in the number of eosinophils and in the levels of IL-4 and IL-5 in bronchoalveolar lavage fluid, compared with those in response to transfer of non-coding plasmid vector or of lipid alone. Conclusion These results suggest that increases in endogenous mST2 protein after allergen exposure may modulate Th2-mediated airway inflammation, and that in vivo gene transfer of mST2 can be applicable to use in a novel immunotherapy for allergic diseases. [source] Efficacy of the Flashlamp-Pumped Pulsed-Dye Laser in Nonsurgical Delay of Skin FlapsDERMATOLOGIC SURGERY, Issue 7 2003Ali Riza Erçöçen MD Objective. The purpose of this article was to determine the effectiveness of laser delay by use of the flashlamp-pumped pulsed-dye laser operating at a wavelength of 585 nm; to elucidate the comparable or dissimilar macroscopic, microscopic, and hemodynamic changes between laser and surgical delay methods; and to clarify the possible mechanisms underlying the delay effect of laser. Methods. A standardized caudally based random dorsal rat flap model was used in this study: Acute random skin flaps served as control subjects (group 1). Surgical delay was employed by incision of lateral longitudinal borders both without (group 2) and with (group 3) undermining, and laser delay methods were performed by laser irradiation of both lateral longitudinal borders (group 4) and the entire surface (group 5) of the proposed flap. Evaluation was done by histologic examination, India ink injection, laser Doppler perfusion imaging, and measurement of flap survival. Results. Histologically, dilation and hypertrophy of subpapillary and subdermal vessels were evident in groups 2, 3, and 4; on the other hand, degranulation of mast cells in the vicinity of occluded vessels at the 1st hour of laser delay and a striking mast cell proliferation and degranulation in association with newly formed vessels (angiogenesis) at the 14th day of laser delay were prominent in group 5. India ink injections revealed longitudinally arranged large-caliber vessels and cross-filling between the vessels of adjacent territories in groups, 2, 3, and 4, but only small-caliber vessels in group 5. Compared with the acute flaps, both surgical and laser delay significantly increased the mean flap perfusion to the maximal levels after a 14-day delay period, and all delay procedures improved flap survival; the most significant increase in surviving area was observed in group 3, whereas the less significant increase in surviving area was in group 5. Conclusion. This study demonstrates that laser delay is as effective as surgical delay and that laser delay by lasering lateral borders leads to dilation and longitudinal rearrangement of the existing vessels rather than angiogenesis, whereas laser delay by lasering the entire surface results in delay effect by inducing angiogenesis due to activation and degranulation of the mast cells. [source] Optimized green fluorescent protein variants provide improved single cell resolution of transgene expression in ascidian embryosDEVELOPMENTAL DYNAMICS, Issue 2 2006Robert W. Zeller Abstract The green fluorescent protein (GFP) is used extensively to monitor gene expression and protein localization in living cells, particularly in developing embryos from a variety of species. Several GFP mutations have been characterized that improve protein expression and alter the emission spectra to produce proteins that emit green, blue, cyan, and yellow wavelengths. DsRed and its variants encode proteins that emit in the orange to red wavelengths. Many of these commercially available fluorescent proteins have been "codon optimized" for maximal levels of expression in mammalian cells. We have generated several fluorescent protein color variants that have been codon optimized for maximal expression in the ascidian Ciona intestinalis. By analyzing quantitative time-lapse recordings of transgenic embryos, we demonstrate that, in general, our Ciona optimized variants are detected and expressed at higher levels than commercially available fluorescent proteins. We show that three of these proteins, expressed simultaneously in different spatial domains within the same transgenic embryo are easily detectable using optimized fluorescent filter sets for epifluorescent microscopy. Coupled with recently developed quantitative imaging techniques, our GFP variants should provide useful reagents for monitoring the simultaneous expression of multiple genes in transgenic ascidian embryos. Developmental Dynamics 235:456,467, 2006. © 2005 Wiley-Liss, Inc. [source] Phosphorylation status of pyruvate dehydrogenase distinguishes metabolic phenotypes of cultured rat brain astrocytes and neuronsGLIA, Issue 10 2010Nader D. Halim Abstract Glucose metabolism in nervous tissue has been proposed to occur in a compartmentalized manner with astrocytes contributing largely to glycolysis and neurons being the primary site of glucose oxidation. However, mammalian astrocytes and neurons both contain mitochondria, and it remains unclear why in culture neurons oxidize glucose, lactate, and pyruvate to a much larger extent than astrocytes. The objective of this study was to determine whether pyruvate metabolism is differentially regulated in cultured neurons versus astrocytes. Expression of all components of the pyruvate dehydrogenase complex (PDC), the rate-limiting step for pyruvate entry into the Krebs cycle, was determined in cultured astrocytes and neurons. In addition, regulation of PDC enzymatic activity in the two cell types via protein phosphorylation was examined. We show that all components of the PDC are expressed in both cell types in culture, but that PDC activity is kept strongly inhibited in astrocytes through phosphorylation of the pyruvate dehydrogenase alpha subunit (PDH,). In contrast, neuronal PDC operates close to maximal levels with much lower levels of phosphorlyated PDH,. Dephosphorylation of astrocytic PDH, restores PDC activity and lowers lactate production. Our findings suggest that the glucose metabolism of astrocytes and neurons may be far more flexible than previously believed. © 2010 Wiley-Liss, Inc. [source] Myelin transcription factor 1 (Myt1) expression in demyelinated lesions of rodent and human CNSGLIA, Issue 7 2007Adam C. Vana Abstract Myelin transcription factor 1 (Myt1) is a zinc-finger DNA binding protein that influences developing oligodendrocyte progenitor (OP) cell proliferation, differentiation, and myelin gene transcription in vitro. The potential of Myt1 to play a role in OP responses leading to remyelination was examined using murine hepatitis virus strain A59 (MHV) to induce spinal cord demyelination and potential relevance to human pathology was evaluated in multiple sclerosis (MS) lesions. In MHV-infected mice, the density of Myt1 expressing cells markedly increased in lesioned areas of spinal cord white matter. Myt1 expressing cells proliferated most extensively during active demyelination and subsequently accumulated to maximal levels during early remyelination. Cells with nuclear Myt1 immunoreactivity were mainly OP cells, identified by co-localization with platelet-derived growth factor alpha receptor, with additional phenotypes being either oligodendrocytes or neural stem cells, identified by CC1 antigen and Musashi1, respectively. The density of OP cells expressing Myt1 was significantly increased in white matter of MHV-infected mice during demyelination and early remyelination then as remyelination advanced the values returned to levels comparable to PBS-injected control mice. In MHV lesions, Myt1 was not expressed in astrocytes, lymphocytes, or macrophage/microglial cells. MS lesions demonstrated increased Myt1 expression in both the periplaque white matter adjacent to lesions and within early remyelinating lesions. These results suggesta potential role for Myt1 in the regeneration of oligodendrocyte lineage cells in response to demyelination. © 2007 Wiley-Liss, Inc. [source] p53 may positively regulate hepatocyte proliferation in ratsHEPATOLOGY, Issue 2 2002Yukiko Inoue p53, known as a tumor suppressor gene, is a transcription factor that regulates various cellular functions. Recently, several growth factor gene promoters, including that of transforming growth factor , (TGF-,), were shown to be direct targets of p53-mediated transcription. Hepatic p53 mRNA is up-regulated during liver regeneration in rats. The aim of this study is to examine the role of p53 in hepatocyte proliferation. p53 protein levels were examined in rat hepatocytes cultured in the medium containing hepatocyte growth factor (HGF). p53 levels began to increase after 6 hours of incubation, reached a maximum at 18 hours, and decreased thereafter. DNA synthesis increased at 12 hours and peaked at 30 hours. When hepatocytes were incubated with p53 antisense oligonucleotide in addition to HGF, increases of p53 and TGF-, levels were suppressed, and DNA synthesis was reduced. The increases of TGF-, levels and DNA synthesis were also suppressed by a chemical inhibitor of p53, pifithrin-,. In rats after two-thirds partial hepatectomy, hepatic p53 increased and reached maximal levels around 16 hours when hepatic HGF levels have been shown to reach a maximum followed by an increase in hepatic TGF-, levels or hepatocyte proliferation. In contrast, sham-operated rats showed minor elevations of hepatic p53 levels. In conclusion, p53 production is stimulated by HGF and may contribute to the proliferation of rat hepatocytes. Considering previous findings indicating the importance of endogenous TGF-, for the proliferation of hepatocytes stimulated by HGF, TGF-, might play a role in HGF-p53 mediated hepatocyte proliferation. [source] Induction of Oxidative DNA Damage in the Peri-Infarct Region After Permanent Focal Cerebral IschemiaJOURNAL OF NEUROCHEMISTRY, Issue 4 2000Tetsuya Nagayama Abstract: To address the role of oxidative DNA damage in focal cerebral ischemia lacking reperfusion, we investigated DNA base and strand damage in a rat model of permanent middle cerebral artery occlusion (MCAO). Contents of 8-hydroxyl-2,-deoxyguanosine (8-OHdG) and apurinic/apyrimidinic abasic sites (AP sites), hallmarks of oxidative DNA damage, were quantitatively measured in nuclear DNA extracts from brains obtained 4-72 h after MCAO. DNA single- and double-strand breaks were detected on coronal brain sections using in situ DNA polymerase I-mediated biotin-dATP nick-translation (PANT) and terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL), respectively. Levels of 8-OHdG and AP sites were markedly elevated 16-72 h following MCAO in the frontal cortex, representing the peri-infarct region, but levels did not significantly change within the ischemic core regions of the caudateputamen and parietal cortex. PANT- and TUNEL-positive cells began to be detectable 4-8 h following MCAO in the caudate-putamen and parietal cortex and reached maximal levels at 72 h. PANT- and TUNEL-positive cells were also detected 16-72 h after MCAO in the lateral frontal cortex within the infarct border, where many cells also showed colocalization of DNA single-strand breaks and DNA fragmentation. In contrast, levels of PANT-positive cells alone were transiently increased (16 h after MCAO) in the medial frontal cortex, an area distant from the infarct zone. These data suggest that within peri-infarct brain regions, oxidative injury to nuclear DNA in the form of base and strand damage may be a significant and contributory cause of secondary expansion of brain damage following permanent focal ischemia. [source] ESTABLISHMENT OF MINIMAL AND MAXIMAL TRANSCRIPT LEVELS FOR NITRATE TRANSPORTER GENES FOR DETECTING NITROGEN DEFICIENCY IN THE MARINE PHYTOPLANKTON ISOCHRYSIS GALBANA (PRYMNESIOPHYCEAE) AND THALASSIOSIRA PSEUDONANA (BACILLARIOPHYCEAE),JOURNAL OF PHYCOLOGY, Issue 4 2009Lee-Kuo Kang Nitrate transporter genes (Nrt2) encode high-affinity nitrate transporters in marine phytoplankton, and their transcript levels are potential markers of nitrogen deficiency in eukaryotic phytoplankton. For the proper interpretation of measured Nrt2 transcript abundances, a relative expression assay was proposed and tested in Isochrysis galbana Parke (Prymnesiophyceae) and Thalassiosira pseudonana (Hust.) Hasle et Heimdal (Bacillariophyceae). The minimal transcript levels of Nrt2 genes were achieved by the addition of 100 ,M ammonium, which led to a rapid decline in Nrt2 transcripts in 10,30 min. Experiments using a concentration series revealed that the effective dosage of ammonium to create a minimal transcript level of ,1 ,mol · mol,1 18S rRNA was ,25 ,M in both species. On the other hand, the addition of l -methionine sulfoximine (MSX), an inhibitor of glutamine synthetase, enhanced the Nrt2 transcript level in I. galbana but did not affect that in T. pseudonana. Nitrogen deprivation was used as an alternative means to create maximal Nrt2 transcript levels. By transferring cells into N-free medium for 24 h, Nrt2 transcript levels increased to ,90 ,mol · mol,1 18S rRNA in I. galbana, and to ,800 ,mol · mol,1 18S rRNA in T. pseudonana. The degree of nitrogen deficiency thus can be determined by comparing original Nrt2 transcript levels with the minimal and maximal levels. [source] Changes in mesophyll anatomy and sink,source relationships during leaf development in Quercus glauca, an evergreen tree showing delayed leaf greeningPLANT CELL & ENVIRONMENT, Issue 5 2003S.-I. MIYAZAWA ABSTRACT Changes in mesophyll anatomy, gas exchange, and the amounts of nitrogen and cell wall constituents including cellulose, hemicellulose and lignin during leaf development were studied in an evergreen broad-leaved tree, Quercus glauca, and in an annual herb, Phaseolus vulgaris. The number of chloroplasts per whole leaf in P. vulgaris increased and attained the maximal level around 10 d before full leaf area expansion (FLE), whereas it continued to increase even after FLE in Q. glauca. The increase in the number of palisade tissue cells per whole leaf continued until a few days before FLE in Q. glauca, but it had almost ceased by 10 d before FLE in P. vulgaris. The radius and height of palisade tissue cells in Q. glauca, attained their maximal levels at around FLE whereas the thickness of the mesophyll cell wall and concentrations of the cell wall constituents increased markedly after FLE. These results clearly indicated that, in Q. glauca, chloroplast development proceeded in parallel with the cell wall thickening well after completion of the mesophyll cell division and cell enlargement. The sink,source transition, defined to be the time when the increase in daily carbon exchange rate exceeds the daily increase in leaf carbon content, occurred before FLE in P. vulgaris but after FLE in Q. glauca. During leaf area expansion, the maximum daily increase in nitrogen content on a whole leaf basis (the maximum leaf areas were corrected to be identical for these species) in Q. glauca was similar to that in P. vulgaris. In Q. glauca, however, more than 70% of nitrogen in the mature leaf was invested during its sink phase, whereas in P. vulgaris it was 50%. These results suggest that Q. glauca invests nitrogen for cell division for a considerable period and for chloroplast development during the later stages. We conclude that the competition for nitrogen between cell division and chloroplast development in the area of expanding leaves can explain different greening patterns among plant species. [source] Dynamics of circulating concentrations of gonadotropins and ovarian hormones throughout the menstrual cycle in the bonnet monkey: role of inhibin A in the regulation of follicle-stimulating hormone secretionAMERICAN JOURNAL OF PRIMATOLOGY, Issue 10 2009P.S. Suresh Abstract In higher primates, increased circulating follicle-stimulating hormone (FSH) levels seen during late menstrual cycle and during menstruation has been suggested to be necessary for initiation of follicular growth, recruitment of follicles and eventually culminating in ovulation of a single follicle. With a view to establish the dynamics of circulating FSH secretion with that of inhibin A (INH A) and progesterone (P4) secretions during the menstrual cycle, blood was collected daily from bonnet monkeys beginning day 1 of the menstrual cycle up to 35 days. Serum INH A levels were low during early follicular phase, increased significantly coinciding with the mid cycle luteinizing hormone (LH) surge to reach maximal levels during the mid luteal phase before declining at the late luteal phase, essentially paralleling the pattern of P4 secretion seen throughout the luteal phase. Circulating FSH levels were low during early and mid luteal phases, but progressively increased during the late luteal phase and remained high for few days after the onset of menses. In another experiment, lutectomy performed during the mid luteal phase resulted in significant decrease in INH A concentration within 2,hr (58.3±2 vs. 27.3±3,pg/mL), and a 2- to 3-fold rise in circulating FSH levels by 24,hr (0.20±0.02 vs. 0.53±0.14,ng/mL) that remained high until 48,hr postlutectomy. Systemic administration of Cetrorelix (150,µg/kg body weight), a gonadotropin releasing hormone receptor antagonist, at mid luteal phase in monkeys led to suppression of serum INH A and P4 concentrations 24,hr post treatment, but circulating FSH levels did not change. Administration of exogenous LH, but not FSH, significantly increased INH A concentration. The results taken together suggest a tight coupling between LH and INH A secretion and that INH A is largely responsible for maintenance of low FSH concentration seen during the luteal phase. Am. J. Primatol. 71:817,824, 2009. © 2009 Wiley-Liss, Inc. [source] Anti-inflammatory properties of heat shock protein 70 and butyrate on Salmonella -induced interleukin-8 secretion in enterocyte-like Caco-2 cellsCLINICAL & EXPERIMENTAL IMMUNOLOGY, Issue 1 2005J. J. Malago Summary Intestinal epithelial cells secrete the chemokine interleukin (IL)-8 in the course of inflammation. Because heat shock proteins (Hsps) and butyrate confer protection to enterocytes, we investigated whether they modulate Salmonella enterica serovar Enteritidis (S. serovar Enteritidis)-induced secretion of IL-8 in enterocyte-like Caco-2 cells. Caco-2 cells incubated with or without butyrate (0,20 m M, 48 h) were infected with S. serovar Enteritidis after (1 h at 42°C, 6 h at 37°C) or without prior heat shock (37°C). Levels of Hsp70 production and IL-8 secretion were analysed using immunostaining of Western blots and enzyme-linked immunosorbent assay (ELISA), respectively. The cells secreted IL-8 in response to S. serovar Enteritidis and produced Hsp70 after heat shock or incubation with butyrate. The IL-8 secretion was inhibited by heat shock and butyrate concentrations as low as 0·2 m M for crypt-like and 1 m M for villous-like cells. In a dose-dependent manner, higher butyrate concentrations enhanced IL-8 secretion to maximal levels followed by a gradual but stable decline. This decline was associated with increasing production of Hsp70 and was more vivid in crypt-like cells. In addition, the higher concentrations abolished the heat shock inhibitory effect. Instead, they promoted the IL-8 production in heat-shocked cells even in the absence of S. serovar Enteritidis. We conclude that heat shock and low concentrations of butyrate inhibit IL-8 production by Caco-2 cells exposed to S. serovar Enteritidis. Higher butyrate concentrations stimulate the chemokine production and override the inhibitory effect of the heat shock. The IL-8 down-regulation could in part be mediated via production of Hsp70. [source] The true cause of pyloric stenosis is hyperacidityACTA PAEDIATRICA, Issue 2 2006Ian Munro Rogers Abstract Pyloric stenosis (PS) has no known cause. A testable theory of cause is proposed, based on the inheritance of a parietal cell mass (PCM) at the upper end of the normal range. It is proposed that, until 3,4 wk of age, the obligatory high fasting gastrins are at maximal levels and not able to be diminished by increasing antral acidity. Hence, rising acidity is not reduced by a lowered gastrin during this time, and very high acidity occurs. Conclusion: Persisting duodenal hyperacidity is created by an inherited high PCM and loss of gastrin control. These two factors produce pyloric stenosis through work hypertrophy from repeated pyloric contraction in response to hyperacidity. [source] | ||||||||||||||||