bluuebunny/crossref_metadata_2025 huggingface dataset:
A revisit of prophylactic lamivudine for chemotherapy-associated hepatitis B reactivation in non-Hodgkin's lymphoma: A randomized trial
Authors: Hsu Chiun , Hsiung Chao A. , Su Ih-Jen , Hwang Wei-Shou , Wang Ming-Chung , Lin Sheng-Fung , Lin Tseng-Hsi , Hsiao Hui-Hua , Young Ji-Hsiung , Chang Ming-Chih , Liao Yu-Min , Li Chi-Cheng , Wu Hung-Bo , Tien Hwei-Fang , Chao Tsu-Yi , Liu Tsang-Wu , Cheng Ann-Lii , Chen Pei-Jer
Published: February 2008

Lamivudine is effective to control hepatitis B virus (HBV) reactivation in HBV-carrying cancer patients who undergo chemotherapy, but the optimal treatment protocol remains undetermined. In this study, HBV carriers with newly diagnosed non-Hodgkin's lymphoma (NHL) who underwent chemotherapy were randomized to either prophylactic (P) or therapeutic (T) lamivudine treatment groups. Group P patients started lamivudine from day 1 of the first course of chemotherapy and continued treatment until 2 months after completion of chemotherapy. Group T patients received chemotherapy alone and started lamivudine treatment only if serum alanine aminotransferase (ALT) levels elevated to greater than 1.5-fold of the upper normal limit (ULN). The primary endpoint was incidence of HBV reactivation during the 12 months after starting chemotherapy. During chemotherapy, fewer group P patients had HBV reactivation (11.5% versus 56%,

P

= 0.001), HBV-related hepatitis (7.7% versus 48%,

P

= 0.001), or severe hepatitis (ALT more than 10-fold ULN) (0 versus 36%,

P

< 0.001). No hepatitis-related deaths occurred during protocol treatment. Prophylactic lamivudine use was the only independent predictor of HBV reactivation. After completion of chemotherapy, the incidence of HBV reactivation did not differ between the 2 groups. Two patients, both in group P, died of HBV reactivation–related hepatitis, 173 and 182 days, respectively, after completion of protocol treatment. When compared with an equivalent group of lamivudine-naïve lymphoma patients who underwent chemotherapy, therapeutic use of lamivudine neither reduced the severity of HBV-related hepatitis nor changed the patterns of HBV reactivation.

Conclusion:

Prophylactic lamivudine use, but not therapeutic use, reduces the incidence and severity of chemotherapy-related HBV reactivation in NHL patients.
Selective portal vein embolization and ligation trigger different regenerative responses in the rat liver
Authors: Furrer Katarzyna , Tian Yinghua , Pfammatter Thomas , Jochum Wolfram , El-Badry Ashraf Mohammad , Graf Rolf , Clavien Pierre-Alain
Published: February 2008

Two strategies are clinically available to induce selective hypertrophy of the liver: portal vein embolization (PVE) and portal vein ligation (PVL). The aim of this study was to compare the impact of PVE and PVL on liver regeneration. Rats were subjected to 70% PVL, 70% PVE, 70% partial hepatectomy (PH) (positive control), or sham operation (negative control). PVL and PVE of liver segments were validated by portography and histology, demonstrating obstruction of the involved portal branches. Liver weight and markers of regeneration were assessed at 24, 48, and 72 hours, and 7 days after surgery (n = 5). Sinusoidal perfusion was examined by intravital microscopy. The weight of the regenerating liver segments increased continuously in all groups, with the highest weight gain after PH, which also disclosed the strongest proliferative activity. In Ki-67 and PCNA stainings, hepatocyte proliferation after PVL was more pronounced than after PVE (

P

= 0.01). Volumetric blood flow and functional sinusoidal density were lower after PVE than after PVL (

P

= 0.006,

P

= 0.02, respectively). The accumulation of Kupffer cells 24 hours after the intervention was highest after PH. Transcript levels of cytokines (interleukin-1β, tumor necrosis factor-α, interleukin-6) peaked at 24 hours and were highest after PH. The embolized part of the liver after PVE showed prominent foreign body reaction in the portal triad with accumulation of macrophages.

Conclusion:

PVL is superior to PVE in inducing a regenerative response of the remnant liver. The impairment of liver regeneration after PVE may be a consequence of macrophage trapping in the occluded segment due to a foreign body reaction. Lower blood flow and lower accumulation of macrophages, particularly Kupffer cells, in the regenerating part of the liver likewise causes impaired liver regeneration after PVE.
The role of Hfe in transferrin-bound iron uptake by hepatocytes
Authors: Chua Anita C.G. , Herbison Carly E. , Drake Sarah F. , Graham Ross M. , Olynyk John K. , Trinder Debbie
Published: February 2008

HFE-related hereditary hemochromatosis results in hepatic iron overload. Hepatocytes acquire transferrin-bound iron via transferrin receptor (Tfr) 1 and Tfr1-independent pathways (possibly Tfr2-mediated). In this study, the role of Hfe in the regulation of hepatic transferrin-bound iron uptake by these pathways was investigated using

Hfe

knockout mice. Iron and transferrin uptake by hepatocytes from

Hfe

knockout, non–iron-loaded and iron-loaded wild-type mice were measured after incubation with 50 nM 125I-Tf-59Fe (Tfr1 pathway) and 5 μM 125I-Tf-59Fe (Tfr1-independent or putative Tfr2 pathway). Tfr1 and Tfr2 messenger RNA (mRNA) and protein expression were measured by real-time polymerase chain reaction and western blotting, respectively. Tfr1-mediated iron and transferrin uptake by

Hfe

knockout hepatocytes were increased by 40% to 70% compared with iron-loaded wild-type hepatocytes with similar iron levels and Tfr1 expression. Iron and transferrin uptake by the Tfr1-independent pathway was approximately 100-fold greater than by the Tfr1 pathway and was not affected by the absence of Hfe. Diferric transferrin increased hepatocyte Tfr2 protein expression, resulting in a small increase in transferrin but not iron uptake by the Tfr1-independent pathway.

Conclusion:

Tfr1-mediated iron uptake is regulated by Hfe in hepatocytes. The Tfr1-independent pathway exhibited a much greater capacity for iron uptake than the Tfr1 pathway but it was not regulated by Hfe. Diferric transferrin up-regulated hepatocyte Tfr2 protein expression but not iron uptake, suggesting that Tfr2 may have a limited role in the Tfr1-independent pathway.
Impaired autophagy: A mechanism of mitochondrial dysfunction in anoxic rat hepatocytes
Authors: Kim Jae-Sung , Nitta Takashi , Mohuczy Dagmara , O'Malley Kerri A. , Moldawer Lyle L. , Dunn William A. , Behrns Kevin E.
Published: February 2008

Autophagy selectively removes abnormal or damaged organelles such as dysfunctional mitochondria. The mitochondrial permeability transition (MPT) is a marker of impaired mitochondrial function that is evident in hepatic ischemia/reperfusion (I/R) injury. However, the relationship between mitochondrial dysfunction and autophagy in I/R injury is unknown. Cultured rat hepatocytes and mouse livers were exposed to anoxia/reoxygenation (A/R) and I/R, respectively. Expression of autophagy-related protein 7 (Atg7), Beclin-1, and Atg12, autophagy regulatory proteins, was analyzed by western blots. Some hepatocytes were incubated with calpain 2 inhibitors or infected with adenoviruses encoding green fluorescent protein (control), Atg7, and Beclin-1 to augment autophagy. To induce nutrient depletion, a condition stimulating autophagy, hepatocytes were incubated in an amino acid–free and serum-free medium for 3 hours prior to onset of anoxia. For confocal imaging, hepatocytes were coloaded with calcein and tetramethylrhodamine methyl ester to visualize onset of the MPT and mitochondrial depolarization, respectively. To further examine autophagy, hepatocytes were infected with an adenovirus expressing green fluorescent protein–microtubule-associated protein light chain 3 (GFP-LC3) and subjected to A/R. Calpain activity was fluorometrically determined with succinyl-Leu-Leu-Val-Tyr-7-amino-4-methylcoumarin. A/R markedly decreased Atg7 and Beclin-1 concomitantly with a progressive increase in calpain activity. I/R of livers also decreased both proteins. However, inhibition of calpain isoform 2, adenoviral overexpression, and nutrient depletion all substantially suppressed A/R-induced loss of autophagy proteins, prevented onset of the MPT, and decreased cell death after reoxygenation. Confocal imaging of GFP-LC3 confirmed A/R-induced depletion of autophagosomes, which was reversed by nutrient depletion and adenoviral overexpression.

Conclusion:

Calpain 2–mediated degradation of Atg7 and Beclin-1 impairs mitochondrial autophagy, and this subsequently leads to MPT-dependent hepatocyte death after A/R.
Granulin-epithelin precursor as a therapeutic target for hepatocellular carcinoma
Authors: Ho Jenny C. , Ip Ying Chi , Cheung Siu Tim , Lee Yuk Ting , Chan Kui Fat , Wong San Yu , Fan Sheung Tat
Published: February 2008

Primary liver cancer, hepatocellular carcinoma (HCC), is the fifth most common cancer and the third leading cancer killer in the world. There is no effective therapeutic option for most HCC patients. A new therapeutic strategy is essential. Granulin-epithelin precursor (GEP, also called progranulin, acrogranin, or PC-derived growth factor) was identified as a potential therapeutic target for HCC from our earlier genome-wide expression profiles. We aimed to conduct a detailed investigation with

in vitro

and animal experiments. We developed the anti-GEP monoclonal antibody (mAb), and examined its effect on hepatoma cells and normal liver cells

in vitro

. A nude mice model transplanted with human HCC was used to investigate if anti-GEP mAb can inhibit tumor growth

in vivo

. We demonstrated that anti-GEP mAb inhibited the growth of hepatoma cells but revealed no significant effect on normal liver cells. In the nude mice model transplanted with human HCC, anti-GEP mAb decreased the serum GEP level and inhibited the growth of established tumors in a dose-dependent manner. The anti-GEP mAb reduced tumor cell proliferation via the p44/42 MAPK and Akt pathways, and reduced tumor angiogenesis to deprive the nutrient supply with reduced microvessel density and tumor vascular endothelial growth factor level.

Conclusion:

We have shown that anti-GEP antibody can inhibit HCC growth, providing evidence that GEP is a therapeutic target for HCC treatment.
Comprehensive evaluation of hepatitis B virus reverse transcriptase substitutions associated with entecavir resistance
Authors: Baldick Carl J. , Tenney Daniel J. , Mazzucco Charles E. , Eggers Betsy J. , Rose Ronald E. , Pokornowski Kevin A. , Yu Cheng F. , Colonno Richard J.
Published: February 2008

Virologic resistance emerging during entecavir (ETV) therapy for hepatitis B virus (HBV) requires three substitutions in the viral reverse transcriptase (RT), signifying a high barrier to resistance. Two of these substitutions are associated with lamivudine resistance (LVDr) in the tyrosine-methionine-aspartate-aspartate (YMDD) motif (rtM204V and rtL180M), whereas the other occurs at one or more positions specifically associated with ETV resistance (ETVr): rtT184, rtS202, or rtM250. Although a variety of substitutions at these primary ETVr positions arise during ETV therapy, only a subset give rise to clinical virologic breakthrough. To determine the phenotypic impact of observed clinical and potential new ETVr substitutions, a comprehensive panel of clones containing every possible amino acid at the three primary ETVr positions in LVDr HBV was constructed and analyzed

in vitro

. A range of replication capacities was observed for the panel, but none of the mutations rescued replication of the LVDr mutant to the wild-type level. More clones with residue rtS202 substitutions were severely impaired than those at rtT184 or rtM250. A wide variety of ETV susceptibilities was observed, ranging from approximately eight-fold (no increase over the LVDr parent) to greater than 400-fold over the wild-type. A correlation was identified between clinically observed substitutions and those displaying higher

in vitro

replication and resistance, especially those from virologic breakthrough patients.

Conclusion:

The high number of tolerated and resistant ETVr substitutions is consistent with models predicting that the mechanism for ETVr is through enhancement of LVDr changes in the RT deoxyribonucleotide triphosphate (dNTP)-binding pocket.
The hepatic stem cell niche: Identification by label-retaining cell assay
Authors: Kuwahara Reiichiro , Kofman Alexander V. , Landis Charles S. , Swenson Scott E. , Barendswaard Els , Theise Neil D.
Published: February 2008

Label retention assays remain the state-of-the-art approach to identify the location of intraorgan epithelial stem cell niches,

in situ

and

in vivo

. They are commonly used in organs with rapid cell turnover but have not been applied to the liver, where cell turnover is very slow. We used a sublethal dose of acetaminophen administered coincident with bromodeoxyuridine to load possible hepatic stem cells in mice with label and then administered a second, sublethal chase of acetaminophen to accomplish “washout” of label from transit amplifying cell populations.

Conclusion:

Four possible hepatic stem cell niches are identified by this approach: the canal of Hering (proximal biliary tree), intralobular bile ducts, periductal “null” mononuclear cells, and peribiliary hepatocytes. These results confirm several different and often contradictory lines of investigation regarding the intrahepatic location of stem/progenitor cells and suggest that the liver has a multi-tiered, flexible system of regeneration rather than a single stem/progenitor cell location.
Hepatocytes can induce death of contacted cells via perforin-dependent mechanism
Authors: Guy Clifford S. , Rankin Sherri L. , Wang Jinguo , Michalak Tomasz I.
Published: February 2008

The liver displays unique immunological properties including the ability to remove aberrant cells and pathogens and to induce peripheral immunotolerance. We have previously demonstrated that hepatocytes can cause cell death by a CD95 ligand–mediated mechanism. Here, we provide evidence that hepatocytes can kill other cells via a perforin-dependent pathway. Using cultured woodchuck hepatocytes and human liver cells as well as freshly isolated woodchuck, mouse, and human hepatocytes, we show that hepatocyte-mediated death of CD95-deficient target cells requires microtubule polymerization, a feature of the granule exocytosis–mediated cytotoxicity. Neutralizing anti-perforin antibodies and short-hairpin RNA directed against perforin messenger RNA confirmed the involvement of perforin in hepatocyte-mediated cell killing.

Conclusion:

This study shows that hepatocytes express biologically competent perforin capable of killing susceptible cells and emphasizes the role of hepatocytes as cytotoxic effectors. This also is the first demonstration of perforin in a non-lymphoid cell type.
Virological and immunological determinants of intrahepatic virus-specific CD8+ T-cell failure in chronic hepatitis C virus infection
Authors: Neumann-Haefelin Christoph , Timm Jörg , Spangenberg Hans Christian , Wischniowski Natalie , Nazarova Natalja , Kersting Nadine , Roggendorf Michael , Allen Todd M. , Blum Hubert E. , Thimme Robert
Published: February 2008

Virus-specific CD8+ T-cells play an important role in the outcome of acute hepatitis C virus (HCV) infection. In the chronic phase, however, HCV can persist despite the presence of virus-specific T-cell responses. Therefore, we set out to perform a full-breadth analysis of the intrahepatic virus-specific CD8+ T-cell response, its relation to the peripheral T-cell response, and the overall influence of viral escape and the genetic restriction on intrahepatic CD8+ T-cell failure. Intrahepatic and peripheral CD8+ T-cells from 20 chronically HCV infected patients (genotype 1) were comprehensively analyzed using overlapping peptides spanning the entire HCV polyprotein in concert with autologous viral sequences that were obtained for all targeted regions. HCV-specific CD8+ T-cell responses were detectable in most (90%) chronically HCV-infected patients, and two thirds of these responses targeted novel previously undescribed epitopes. Most of the responses were detectable only in the liver but not in the peripheral blood, indicating accumulation and enrichment at the site of disease. Of note, only approximately half of the responses were associated with viral sequence variations supported by functional analysis as viral escape mutations. Escape mutations were more often associated with HLA-B alleles.

Conclusion:

Our results show an unexpected high frequency of intrahepatic virus-specific CD8+ T-cells, a large part of which continue to target the present viral antigens. Thus, our results suggest that factors other than mutational escape contribute to the failure of intrahepatic virus-specific CD8+ T-cells.
Hepatitis C virus infection: Molecular pathways to metabolic syndrome
Authors: Sheikh Muhammad Y. , Choi Jinah , Qadri Ishtiaq , Friedman Jacob E. , Sanyal Arun J.
Published: February 2008

Chronic infection with hepatitis C virus (HCV) can induce insulin resistance (IR) in a genotype-dependent fashion, thus contributing to steatosis, progression of fibrosis and resistance to interferon therapy. The molecular mechanisms in genotype 1 patients that lead to metabolic syndrome are still ambiguous. Based on our current understanding, HCV proteins associate with mitochondria and endoplasmic reticulum and promote oxidative stress. The latter mediates signals involving the p38 mitogen-activated protein kinase and activates nuclear factor kappa B. This transcription factor plays a key role in the expression of cytokines, tumor necrosis factor alpha (TNF-α), interleukin 6, interleukin 8, tumor growth factor beta, and Fas ligand. TNF-α inhibits the function of insulin receptor substrates and decreases the expression of the glucose transporter and lipoprotein lipase in peripheral tissues, which is responsible for the promotion of insulin resistance. Furthermore, reduced adiponectin levels, loss of adiponectin receptors, and decreased anti-inflammatory peroxisome proliferator-activated receptor alpha in the liver of HCV patients may contribute to reduced fatty acid oxidation, inflammation, and eventually lipotoxicity. This chain of events may be initiated by HCV-associated IR and provides a direction for future research in the areas of therapeutic intervention.
Reactive polyurethane carbon nanotube foams and their interactions with osteoblasts
Authors: Verdejo Raquel , Jell Gavin , Safinia Laleh , Bismarck Alexander , Stevens Molly M. , Shaffer Milo S.P.
Published: February 2008

The remarkable intrinsic properties of carbon nanotubes, including their high mechanical strength, electrical conductivity, and nanoscale 3D architecture, create promising opportunities for the use of nanotube composites in a number of fields, particularly for composites in which conventional fillers cannot be accommodated. In the current study, 3D polyurethane (PU) nanocomposite foams were developed, and their potential biomedical applications were investigated. Multiwalled carbon nanotubes (CNTs) were synthesized by chemical vapor deposition and, following suitable chemical modification, uniformly distributed within the walls of PU foams produced by direct reaction. Although the loading fraction was too low to observe significant mechanical effects, CNT incorporation improved the wettability of the nanocomposite surfaces in a concentration‐dependent manner, supporting the claim that the nanotubes are active at the pore surface. Studies of bone cell interactions with the nanocomposite foams revealed that increasing CNT loading fraction did not cause osteoblast cytotoxicity nor have any detrimental effects on osteoblast differentiation or mineralization. The application of “fixed” or embedded CNTs in nondegradable scaffolds is likely advantageous over “loose” or unattached CNTs from a toxicological point of view. © 2008 Wiley Periodicals, Inc. J Biomed Mater Res, 2009
Effects of bone morphogenetic protein on neonatal rat calvarial osteoblast‐like cells: An in vitro study
Authors: Aybar Buket , Emes Yusuf , Atalay Belir , Vural Pervin , Kaya A. Selhan , Eren Selame N. , İşsever Halim , Bilir Ayhan
Published: February 2008

Bone morphogenetic proteins (BMPs) are factors that promote osteoblastic differentiation and osteogenesis. The aim of this study was to examine the behavior of neonatal rat calvarial osteoblast cells cultured on different concentrations of BMP graft materials. Fifty thousand cells per milliliter were seeded and cultured on graft materials for 24 and 48 h. Different concentrations of BMPs (combination of BMPs numbered from 1 to 14) were supplemented to the medium. To evaluate cellular proliferation and differentiation, specimens were examined for DNA synthesis, alkaline phosphatase (ALP) activity, cell numbers, and viability of the cells. Further, transforming growth factor‐β1 (TGF‐β1) and lactate dehydrogenase (LDH) levels were investigated. Morphological appearance of the specimens at 24 and 48 h of incubation was evaluated using scanning electron microcopy. Evaluations of DNA synthesis, cell count, and cell viability data revealed that a significant difference existed at 24 and 48 h (p < 0.05). The TGF‐β1 and ALP analysis showed only a significant difference between the groups at the end of 24 h (p < 0.05). Regarding the lactate dehydrogenase activity there was not any significant difference at 24 and 48 h (p > 0.05). No morphological differences were observed in cell morphology on BMP graft material and the control group. These results indicate that BMPs have an inductive effect on osteoblast differentiation and a possible inhibitory effect in the early phases of cell proliferation. © 2008 Wiley Periodicals, Inc. J Biomed Mater Res, 2008
APT102, a novel adpase, cooperates with aspirin to disrupt bone metastasis in mice
Authors: Uluçkan Özge , Eagleton Mark C. , Floyd Desiree H. , Morgan Elizabeth A. , Hirbe Angela C. , Kramer Matthew , Dowland Nikki , Prior Julie L. , Piwnica‐Worms David , Jeong Soon Seog , Chen Ridong , Weilbaecher Katherine
Published: February 2008

Platelets contribute to the development of metastasis, the most common cause of mortality in cancer patients, but the precise role that anti‐platelet drugs play in cancer treatment is not defined. Metastatic tumor cells can produce platelet αIIbβ3 activators, such as ADP and thromboxane A2 (TXA2). Inhibitors of platelet β3 integrins decrease bone metastases in mice but are associated with significant bleeding. We examined the role of a novel soluble apyrase/ADPase, APT102, and an inhibitor of TXA2 synthesis, acetylsalicylic acid (aspirin or ASA), in mouse models of experimental bone metastases. We found that treatment with ASA and APT102 in combination (ASA + APT102), but not either drug alone, significantly decreased breast cancer and melanoma bone metastases in mice with fewer bleeding complications than observed with αIIbβ3 inhibition. ASA + APT102 diminished tumor cell induced platelet aggregation but did not directly alter tumor cell viability. Notably, APT102 + ASA treatment did not affect initial tumor cell distribution and similar results were observed in β3−/− mice. These results show that treatment with ASA + APT102 decreases bone metastases without significant bleeding complications. Anti‐platelet drugs such as ASA + APT102 could be valuable experimental tools for studying the role of platelet activation in metastasis as well as a therapeutic option for the prevention of bone metastases. J. Cell. Biochem. 104: 1311–1323, 2008. © 2008 Wiley‐Liss, Inc.
Cold‐triggered/heat‐destroyed emulsions composed of phospholipids and triacylglycerols as thermal history indicators for cold‐chain distribution systems
Authors: Nagata Kohdai , Mizoguchi Tatsuya , Kitsunai Makoto , Hirose Kumi , Hirasawa Akira , Chiba Kazuhiro
Published: February 2008

BACKGROUND: Recently, cold‐chain distribution systems have come to play important roles in worldwide food processing/storage/transportation networks. To ensure the maintenance of the quality and safety of foods, it is necessary to develop thermal history indicators for products involved in cold‐chain distribution systems. To provide a record of the occurrence of a high‐temperature event during a cold‐chain system, a temperature‐related phase change in a material within the indicator is needed to indicate that the high‐temperature event occurred. Preferentially, for safety, the materials of the indicators should be edible and easy to handle. It should be possible to store the indicators at ambient temperature before use, and they should be triggered automatically just by cooling at the start of a cold‐chain system. Furthermore, if the indicator is heated even once during the cold‐chain distribution system, its appearance must be irreversibly altered to provide evidence of the high‐temperature experience.

RESULTS: Based on the edible materials composed of lyso‐lecithin (15 g), lecithin (2 g), triacylglycerol (150 g), and water, we successfully constructed a stable emulsion that could be triggered just by cooling to 4 °C or lower for more than 12 h. After triggering, it was immediately destroyed by heating up to 20 °C or higher. Furthermore, the mechanism of cold‐triggering and heat‐destruction has been studied by nuclear magnetic resonance.

CONCLUSION: The cold‐triggered/heat‐destroyed emulsion should be applied as a new thermal history indicator that can be automatically triggered just by cooling down and irreversibly change its appearance after a high‐temperature experience in cold‐chain distribution systems. Copyright © 2008 Society of Chemical Industry
Endotoxemia does not limit energy supply in exercising rat skeletal muscle
Authors: Giannesini Benoit , Izquierdo Marguerite , Dalmasso Christiane , Fur Yann Le , Cozzone Patrick J. , Verleye Marc , Guern Marie‐Emmanuelle Le , Gillardin Jean‐Marie , Bendahan David
Published: February 2008

Although depletion in high‐energy phosphorylated compounds and mitochondrial impairment have been reported in septic skeletal muscle at rest, their impact on energy metabolism has not been documented during exercise. In this study we aimed to investigate strictly gastrocnemius muscle function non‐invasively, using magnetic resonance techniques in endotoxemic rats. Endotoxemia was induced by injecting animals intraperitoneally at t0 and t0 + 24 h with Klebsiella pneumoniae lipopolysaccharides (at 3 mg kg−1). Investigations were performed at t0 + 48 h during a transcutaneous electrical stimulation protocol consisting of 5.7 min of repeated isometric contractions at a frequency of 3.3 HZ. Endotoxin treatment produced a depletion in basal phosphocreatine content and a pronounced reduction in oxidative adenosine triphosphate (ATP) synthesis capacity, whereas the resting ATP concentration remained unchanged. During the stimulation period, endotoxemia caused a decrease in force‐generating capacity that was fully accounted for by the loss of muscle mass. It further induced an acceleration of glycolytic ATP production and an increased accumulation of adenosine diphosphate (ADP, an important mitochondrial regulator) that allowed a near‐normal rate of oxidative ATP synthesis. Finally, endotoxemia did not affect the total rate of ATP production or the ATP cost of contraction throughout the whole stimulation period. These data demonstrate that, in an acute septic phase, metabolic alterations in resting muscle do not impact energy supply in exercising muscle, likely as a result of adaptive mechanisms. Muscle Nerve, 2008
A comprehensive genetic profile of phyllodes tumours of the breast detects important mutations, intra‐tumoral genetic heterogeneity and new genetic changes on recurrence
Authors: Jones AM , Mitter R , Springall R , Graham T , Winter E , Gillett C , Hanby AM , Tomlinson IPM , Sawyer EJ
Published: February 2008

The aims of this study were to identify genetic changes associated with malignant progression of the fibroepithelial neoplasms, phyllodes tumours of the breast (PTs), and to ascertain whether genetic progression occurs when PTs recur locally. A further aim was to assess whether the genetic data support the classification of these tumours into three subtypes, benign, borderline and malignant. 126 PTs (37 benign, 41 borderline, 48 malignant) were analysed by either array‐CGH or the Illumina Goldengate assay. The large‐scale genetic changes associated with malignant/borderline phenotypes were + 1q, + 5p, + 7, + 8, − 6, − 9p, − 10p and − 13. Cluster analysis of the array‐CGH data supported the division of malignant and borderline PTs into two separate groups, one comprising almost all malignant lesions and the other, benign and borderline tumours. Interstitial deletions of 9p21 that involved the p16INK4a locus were present in many malignant/borderline PTs, and some of these appeared to cause homozygous loss. Loss of expression of p16INK4a was found frequently and this was associated with 9p deletion; we also identified one p16INK4a mutation and evidence of methylation of p16INK4a in malignant PTs. Our evidence shows that inactivation of this gene is important in the development of malignant PTs. In selected PTs, multiple areas of stroma were isolated and analysed separately by array‐CGH. We found considerable intra‐tumoral genetic heterogeneity. Analysis of paired primary and recurrent tumours showed that recurrent tumours often acquired new genetic changes; in particular, benign tumours tended to acquire changes characteristic of the malignant/borderline phenotype. We believe it likely that unfavourable sub‐clones not easily identified by histology account for the unpredictable clinical behaviour of these tumours. Copyright © 2008 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.
The RNA‐binding and adaptor protein Sam68 modulates signal‐dependent splicing and transcriptional activity of the androgen receptor
Authors: Rajan P , Gaughan L , Dalgliesh C , El‐Sherif A , Robson CN , Leung HY , Elliott DJ
Published: February 2008

The RNA‐binding protein Sam68 has been reported to be up‐regulated in clinical cases of prostate cancer (PCa), where it is thought to contribute to cell proliferation and survival. Consistent with this, we observed over‐expression of Sam68 in a panel of clinical prostate tumours as compared with benign controls. Since Sam68 is implicated in a number of signalling pathways, we reasoned that its role in PCa may involve modulation of the androgen receptor (AR) signalling cascade, which drives the onset and progression of PCa. We found that Sam68 interacts with the AR in vivo in LNCaP cells, and is dynamically recruited to androgen response elements within the promoter region of the prostate‐specific antigen (PSA) gene. Based on its known functions and nuclear location, Sam68 might either: (a) co‐regulate AR‐dependent transcription positively or negatively; or (b) modulate AR‐dependent alternative splicing by enhancing incorporation of a Sam68‐responsive exon transcribed under the control of an androgen‐responsive promoter. We tested these possibilities using functional assays. Both wild‐type Sam68 protein and the Sam68V229F mutant, which is impaired in RNA binding, functioned as a ligand‐dependent AR co‐activator on an androgen‐regulated reporter gene. In contrast, splicing of a Sam68‐responsive variable exon, transcribed under control of an androgen‐responsive promoter, was strongly repressed in the presence of AR and androgens. This splicing inhibition was reversed by ectopic expression of Sam68 but enhanced by Sam68V229F. These results demonstrate that Sam68 has separable effects on AR‐regulated transcriptional activity and alternative splicing, both of which may affect PCa phenotypes. Copyright © 2008 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.
Functional and phenotypic characteristics of testicular macrophages in experimental autoimmune orchitis
Authors: Rival C , Theas MS , Suescun MO , Jacobo P , Guazzone V , van Rooijen N , Lustig L
Published: February 2008

Testicular inflammation with compromised fertility can occur despite the fact that the testis is considered an immunoprivileged organ. Testicular macrophages have been described as cells with an immunosuppressor profile, thus contributing to the immunoprivilege of the testis. Experimental autoimmune orchitis (EAO) is a model of organ‐specific autoimmunity and testicular inflammation. EAO is characterized by an interstitial inflammatory mononuclear cell infiltration, damage of the seminiferous tubules and germ cell apoptosis. Here we studied the phenotype and functions of testicular macrophages during the development of EAO. By stereological analysis, we detected an increased number of resident (ED2+) and non‐resident (ED1+) macrophages in the testicular interstitium of rats with orchitis. We showed that this increase was mainly due to monocyte recruitment. The in vivo administration of liposomes containing clodronate in rats undergoing EAO led to a reduction in the number of testicular macrophages, which correlated with a decreased incidence and severity of the testicular damage and suggests a pathogenic role of macrophages in EAO. By immunohistochemistry and flow cytometry we detected an increased number of testicular macrophages expressing MHC class II, CD80 and CD86 costimulatory molecules in rats with orchitis. Also, testicular macrophages from rats with EAO showed a higher production of IFNγ (ELISA). We conclude that testicular macrophages participate in EAO development, and the ED1+ macrophage subset is the main pathogenic subpopulation. They stimulate the immune response through the production of pro‐inflammatory cytokines and antigen presentation and thus activation of T cells in the target organ. Copyright © 2008 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.
The androgen‐regulated type II serine protease TMPRSS2 is differentially expressed and mislocalized in prostate adenocarcinoma
Authors: Lucas JM , True L , Hawley S , Matsumura M , Morrissey C , Vessella R , Nelson PS
Published: February 2008

Transmembrane serine protease 2 (TMPRSS2) is an androgen‐regulated member of the type two transmembrane protease (TTSP) family. Two other members of the TTSP family, matriptase and hepsin, are over‐expressed in prostate adenocarcinoma and mechanistically influence cancer cell invasion and metastasis. This study was performed to determine TMPRSS2 protein expression in primary and metastatic prostate cancers. We developed a monoclonal antibody capable of the sensitive and specific detection of TMPRSS2 protein. TMPRSS2 regulation by androgen and presence in seminal fluid was measured. TMPRSS2 localization and expression was evaluated in 415 cases of primary prostate cancer and 144 prostate cancer metastases by immunohistochemistry. We determined that TMPRSS2 protein expression is regulated by androgens and that TMPRSS2 is a component of the normal seminal fluid proteome. TMPRSS2 protein is abundantly expressed in the prostate, with low levels in the epithelia of the colon, stomach, epididymis and breast. Pancreatic acini, hepatic bile ducts, testicular Leydig cells and the kidney also express TMPRSS2. In the prostate, TMPRSS2 protein is specifically localized to the secretory epithelium, with enhanced expression in the plasma membrane orientated towards the ductal lumen. TMPRSS2 expression was significantly higher in both neoplastic prostate and in the epithelium of prostatic hyperplasia compared to normal epithelium (p < 0.01). TMPRSS2 expression was further elevated in higher Gleason grade cancers (patterns 4 and 5) compared to pattern 3 (p = 0.04). Furthermore, in most high‐grade cancers, TMPRSS2 was mislocalized, being expressed in the cytoplasm as well as in the cell membrane. Prostate cancer metastases also generally expressed high levels of TMPRSS2. In summary, the TMPRSS2 protease is expressed highly in primary and metastatic prostate cancers and is associated with tumour cell differentiation. Based on studies with the related proteins matriptase and hepsin, TMPRSS2 should be investigated for causal roles in prostate carcinogenesis. Copyright © 2008 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.
Initial testing (stage 1) of a monoclonal antibody (SCH 717454) against the IGF‐1 receptor by the pediatric preclinical testing program
Authors: Kolb E. Anders , Gorlick Richard , Houghton Peter J. , Morton Christopher L. , Lock Richard , Carol Hernan , Reynolds C. Patrick , Maris John M. , Keir Stephen T. , Billups Catherine A. , Smith Malcolm A.
Published: February 2008

Background

SCH 717454 (19D12) is a fully human antibody directed against the insulin‐like growth factor 1 receptor (IGF‐1R), which is implicated in the growth and metastatic phenotype of a broad range of malignancies. The activity of SCH 717454 was evaluated against the in vitro and in vivo panels of the Pediatric Preclinical Testing Program (PPTP).

Procedures

SCH 717454 was tested against the PPTP in vitro panel at concentrations ranging from 0.01 to 100 nM and was tested against the PPTP in vivo panel at a dose of 0.5 mg per mouse administered twice weekly for 4 weeks via intraperitoneal injection.

Results

SCH 717454 was ineffective at retarding growth of cell lines in the in vitro panel. In vivo, SCH 717454 significantly increased event‐free survival in 20 of 35 (57%) solid tumor xenograft models with tumor regressions in one Ewing sarcoma model (complete response) and 2 osteosarcoma models (maintained complete responses). Using the time to event activity measure, SCH 717454 had intermediate (n = 9) or high (n = 1) activity against 31 evaluable solid tumor xenografts, including xenografts from the rhabdoid tumor, Ewing, rhabdomyosarcoma, glioblastoma, neuroblastoma, and osteosarcoma panels. SCH 717454 showed little activity against the 8 xenografts of the acute lymphoblastic leukemia panel.

Conclusions

SCH 717454 demonstrated broad antitumor activity against the PPTP's in vivo solid tumor panels. Further characterization of the molecular predictors of response and of the activity of combinations of SCH 717454 with other anticancer agents are anticipated. Pediatr Blood Cancer 2008;50:1190–1197. © 2008 Wiley‐Liss, Inc.
Mycoplasma pneumoniae and atypical Stevens–Johnson syndrome in a hematopoietic stem cell transplant recipient
Authors: Birch Jennifer , Chamlin Sarah , Duerst Reggie , Jacobsohn David
Published: February 2008

Stevens–Johnson syndrome (SJS) is not typically reported following hematopoietic stem cell transplant (HSCT). The most severe form of SJS, which is toxic epidermal necrolysis (TEN) has been reported following HSCT, albeit very rarely. We describe a case of Mycoplasma‐associated SJS following HSCT. While this association is commonly reported in previously healthy children, it has not been reported in patients following HSCT. Pediatr Blood Cancer 2008;50:1278–1279. © 2008 Wiley‐Liss, Inc.
Descriptive epidemiology of childhood Langerhans cell histiocytosis in France, 2000–2004
Authors: Guyot‐Goubin Aurélie , Donadieu Jean , Barkaoui Mohamed , Bellec Stéphanie , Thomas Caroline , Clavel Jacqueline
Published: February 2008

Introduction

Childhood Langerhans cell histiocytosis (LCH) is a rare and poorly understood multisystemic disease. The French National Registry of Childhood Hematopoietic Malignancies (NRCH) has recorded LCH cases of all subtypes since 2000. The present study describes the data on LCH collected on a national scale over a 5‐year period.

Materials and Methods

The cases were children aged less than 15 years, diagnosed with LCH of any type between 2000 and 2004, and residing in mainland France at the time of diagnosis. Completeness was evaluated by capture–recapture after cross‐checking against the database compiled by the French Langerhans Cell Histiocytosis Study Group.

Results

Two hundred fifty‐eight cases of LCH were registered. The completeness of the NRCH was estimated to be 97%. The annual incidence rate was 4.6/106 children aged less than 15 years and the sex ratio was 1.2. Bone and skin were the most commonly involved organs at diagnosis. The incidence rate decreased with age from 15.3/106 before 1 year to 2.0/106 after 10 years. The disease was mainly unifocal (2.6/106) and rarely disseminated (0.6/106), but disseminated forms predominated in infants. The overall 2‐year survival rate was 99% (95%CI: [97; 100]). About 30% of the LCH cases were enrolled in a clinical trial at first onset. No case was treated by radiotherapy.

Conclusion

This study evidenced the main features of LCH incidence in the overall population and was consistent with previous studies. The NRCH thus appears to be a very promising tool for further elucidation of LCH. Pediatr Blood Cancer 2008;51:71–75. © 2008 Wiley‐Liss, Inc.
A New Concept to Characterize Nonspherical Particles from Multi‐wavelength Depolarization Ratios Based on T‐matrix Computation
Authors: Somekawa Toshihiro , Yamanaka Chihiro , Fujita Masayuki , Galvez Maria Cecillia
Published: February 2008

A new concept to derive particle information from depolarization ratios obtained by a simultaneous three‐wavelength depolarization lidar was presented. This new data treatment utilized the wavelength dependence of the ratio of the polarization components in the scattering light based on T‐matrix computation. The results showed that calculated three‐wavelength depolarization ratios can be used to characterize particle sizes of ice particles. Assessing the mixing of spherical and nonspherical aerosols, the presented method successfully reproduced the lidar data in real.
Influence of the germline sequence on the thermodynamic stability and fibrillogenicity of human lambda 6 light chains
Authors: del Pozo Yauner Luis , Ortiz Ernesto , Sánchez Rosalba , Sánchez‐López Rosana , Güereca Leopoldo , Murphy Charles L. , Allen Amy , Wall Jonathan S. , Fernández‐Velasco D. Alejandro , Solomon Alan , Becerril Baltazar
Published: February 2008

Light chain‐associated amyloidosis is a fatal disease characterized by the aggregation and pathologic deposition of monoclonal light chain‐related fragments as amyloid fibrils in organs or tissues throughout the body. Notably, it has been observed that proteins encoded by the λ variable light chain (VL) gene segment 6a are invariably associated with amyloid deposition; however, the contribution of the gene to this phenomenon has not been established. In this regard, we have determined the thermodynamic stability and kinetics of in vitro fibrillogenesis of a recombinant (r) VLprotein, designated 6aJL2, which contains the predicted sequences encoded by the 6a and JL2 germline genes. Additionally, we studied a 6a mutant (6aJL2‐Arg25Gly), that is present in ∼25% of all amyloid‐associated λ6 light chains. Remarkably, the wild‐type 6aJL2 protein was more stable than were all known amyloidogenic κ and λ light chains for which stability parameters are available; more importantly, it was even more so (and less fibrillogenic) than the only clinically proven nonamyloidogenic λ6 protein, Jto. Conversely, the mutated 6aJL2‐R25G molecule was considerably less stable and more fibrillogenic than was the native 6aJL2. Our data indicate that the propensity of λ6 light chains to form amyloid can not be attributed to thermodynamic instability of the germline‐encoded Vλ6 domain, but rather, is dependent on sequence alterations that render such proteins amyloidogenic. Proteins 2008. © 2008 Wiley‐Liss, Inc.
PSLDoc: Protein subcellular localization prediction based on gapped‐dipeptides and probabilistic latent semantic analysis
Authors: Chang Jia‐Ming , Su Emily Chia‐Yu , Lo Allan , Chiu Hua‐Sheng , Sung Ting‐Yi , Hsu Wen‐Lian
Published: February 2008

Prediction of protein subcellular localization (PSL) is important for genome annotation, protein function prediction, and drug discovery. Many computational approaches for PSL prediction based on protein sequences have been proposed in recent years for Gram‐negative bacteria. We present PSLDoc, a method based on gapped‐dipeptides and probabilistic latent semantic analysis (PLSA) to solve this problem. A protein is considered as a term string composed by gapped‐dipeptides, which are defined as any two residues separated by one or more positions. The weighting scheme of gapped‐dipeptides is calculated according to a position specific score matrix, which includes sequence evolutionary information. Then, PLSA is applied for feature reduction, and reduced vectors are input to five one‐versus‐rest support vector machine classifiers. The localization site with the highest probability is assigned as the final prediction. It has been reported that there is a strong correlation between sequence homology and subcellular localization (Nair and Rost, Protein Sci 2002;11:2836–2847; Yu et al., Proteins 2006;64:643–651). To properly evaluate the performance of PSLDoc, a target protein can be classified into low‐ or high‐homology data sets. PSLDoc's overall accuracy of low‐ and high‐homology data sets reaches 86.84% and 98.21%, respectively, and it compares favorably with that of CELLO II (Yu et al., Proteins 2006;64:643–651). In addition, we set a confidence threshold to achieve a high precision at specified levels of recall rates. When the confidence threshold is set at 0.7, PSLDoc achieves 97.89% in precision which is considerably better than that of PSORTb v.2.0 (Gardy et al., Bioinformatics 2005;21:617–623). Our approach demonstrates that the specific feature representation for proteins can be successfully applied to the prediction of protein subcellular localization and improves prediction accuracy. Besides, because of the generality of the representation, our method can be extended to eukaryotic proteomes in the future. The web server of PSLDoc is publicly available at http://bio‐cluster.iis.sinica.edu.tw/∼bioapp/PSLDoc/. Proteins 2008. © 2008 Wiley‐Liss, Inc.
Mutant bovine odorant‐binding protein: Temperature affects the protein stability and dynamics as revealed by infrared spectroscopy and molecular dynamics simulations
Authors: Marabotti Anna , Lefèvre Thierry , Staiano Maria , Crescenzo Roberta , Varriale Antonio , Rossi Mosè , Pézolet Michel , D'Auria Sabato
Published: February 2008

Bovine odorant‐binding protein (bOBP), a member of the lipocalin family, presents the so‐called 3D “domain‐swapped” protein structure. In fact, in solution, it appears as a dimer in which each monomer is composed by the classical lipocalin fold, with a central β‐barrel followed by a stretch of residues and the α‐helix domain protruding out of the barrel and crossing the dimer interface. Recently, a deswapped mutant form of bOBP was obtained, in which a Gly residue was inserted after position 121 and the two residues in position 64 and 156 were replaced by Cys residues for restoring the disulfide bridge common to the lipocalin family. In this work, we used Fourier transform infrared spectroscopy and molecular dynamics simulations to investigate the effect of temperature on the structural stability and conformational dynamics of the mutant bOBP. The spectroscopic and molecular simulation data pointed out that the hydrophobic regions of the protein matrix appear to be an important factor for the protein stability and integrity. In addition, it was also found that the mutant bOBP is significantly stabilized by the binding of the ligand, which may have an impact on the biological function of bOBP. The obtained results will allow for a better use of this protein as probe for the design of advanced protein‐based biosensors for the detection of compounds used in the fabrication of explosive powders. Proteins 2008. © 2008 Wiley‐Liss, Inc.