bluuebunny/crossref_metadata_2025 huggingface dataset:
Should Heart Failure Be Treated with Antithrombotic Drugs?
Authors: Sosin Michael , Bhatia Gurbir , Lip Gregory Y.H.
Published: April 2003

Congestive heart failure (CHF) is increasing in prevalence in Western society and is associated with considerable morbidity and mortality, including a high risk of venous thromboembolism and stroke. In addition, up to 50% of CHF sufferers die of sudden cardiac death, of which a major mechanism may be thrombotic coronary occlusion leading to myocardial infarction. Thus, antithrombotic therapy would be expected to provide a substantial morbidity and mortality benefit to these patients. However, clinical evidence to support this hypothesis is limited and inconsistent. Aspirin is routinely used in patients with CHF secondary to ischaemic heart disease; however, as yet, no randomised trials of long-term aspirin therapy in CHF have been reported. Since warfarin therapy is associated with a definite risk of bleeding, anticoagulation should be considered only on a case-by-case basis for patients with CHF and additional risk factors (including atrial fibrillation, previous thromboembolic stroke and mural thrombus formation) until more definitive evidence is available.
Acrylamide: Increased Concentrations in Homemade Food and First Evidence of Its Variable Absorption from Food, Variable Metabolism and Placental and Breast Milk Transfer in Humans
Authors: Sörgel Fritz , Weissenbacher Rainer , Kinzig-Schippers Martina , Hofmann Annette , Illauer Michael , Skott Andreas , Landersdorfer Cornelia
Published: April 2003

We have developed a liquid chromatography/mass spectrometry (LC-MS/MS) assay to determine acrylamide in various body fluids. The assay also allows the reliable quantitation of acrylamide in food. In a total of 11 healthy male and female subjects, we were able to show that acrylamide from food given to humans is in fact absorbed from the gut. The half-lives determined in two male subjects were 2.2 and 7 h. Acrylamide was found in human breast milk and penetrated the human placenta (n = 3). The variability of acrylamide concentrations found in this investigation is most likely caused by variable intersubject bioavailability and metabolism. This may be an important indication that the assessment of the risk from acrylamide for the individual may be very difficult without knowing the concentrations of acrylamide in the body. This should be considered in the design of any risk assessment study or post hoc analysis of earlier studies. At this time, we suggest that pregnant women and breast-feeding mothers avoid acrylamide-containing food.
Expression of HCV E1 Protein in Baculovirus-Infected Cells: Effects on Cell Viability and Apoptosis Induction
Authors: Ciccaglione Anna Rita , Marcantonio Cinzia , Costantino Angela , Equestre Michele , Rapicetta Maria
Published: April 2003

The molecular mechanisms of pathogenesis in hepatitis C virus (HCV) infection are not yet understood. Recently, we reported that the expression of the envelope protein E1 is toxic for Escherichia coli cells. The toxicity is related to the ability of C-terminal transmembrane (TM) domain of E1 to modify membrane permeability. In this study we expressed the E1 protein, complete (a.a. 192–383) or deleted (a.a. 192–340) of the TM region, fused to the C-terminus of glutathione-S-transferase by two recombinant baculoviruses. Infection of Sf9 insect cells by E1 baculovirus induced a rapid decrease in cell viability in the first 18–24 h postinfection. Premature cytopathic changes and low level of E1 protein expression were also reported. The analysis of DNA isolated from cells revealed a typical internucleosomal ladder pattern characteristic of apoptosis. The DNA degradation was first detected at 18 h postinfection by ethidium bromide gel electrophoresis and was confirmed by TUNEL assay. The results indicated that the C-terminal domain of E1 is essential for apoptosis induction as neither cell death nor DNA degradation were observed following infection with the recombinant baculovirus expressing the C-terminal-deleted E1. These findings support the hypothesis that the TM domain of E1 may play a role in viral pathogenesis.
Evaluation and Characterization of Mouse Scratching Behavior by a New Apparatus, MicroAct
Authors: Inagaki N. , Igeta K. , Shiraishi N. , Kim J.F. , Nagao M. , Nakamura N. , Nagai H.
Published: April 2003

We evaluated and characterized the mouse scratching behavior using a new apparatus, MicroAct. Scratching behavior was evoked in ICR and BALB/c mice by compound 48/80, passive cutaneous anaphylaxis or repeated hapten application. Under the present experimental condition, MicroAct detected consecutive scratching behavior (events) consisting of 3 or more beats. Although the detecting standard of MicroAct was not identical to that of an observer, the number of events detected by MicroAct and by an observer were almost comparable with each other. Frequency of events, total scratching time and total number of beats detected by MicroAct increased depending on the intensity of the causing stimuli for scratching. In contrast, the duration of each event and the number of beats in each event increased only slightly, but the scratching speed was almost constant. The present results demonstrate that MicroAct is a useful tool for evaluating mouse scratching behavior. Mouse scratching behavior seems to have a relatively fixed pattern and the causing stimulus increases mainly in the frequency of event without affecting the scratching speed.
EEMCO Guidance for the in vivo Assessment of Skin Surface pH
Authors: Parra J.L. , Paye M.
Published: April 2003

The pH of the skin follows a sharp gradient across the stratum corneum (SC), which is suspected to play an important role in controlling the enzymatic activities involved in cellular metabolism and renewal. This gradient is maintained by several systems, such as sweat and sebum secretion and degradation as well as cellular metabolism. At the surface of the skin, what is measured is in fact an apparent skin pH due to extracted material from the SC diffusing into water applied at the surface. pH values recorded at the surface of a semi hydrophobic milieu such as the SC should be interpreted with great caution because it is obvious that hydrogen ions are not in a pure solution at the surface of the skin. For a correct measurement of skin surface pH, it is recommended to follow all practical operating conditions. Care must be taken in identifying the skin site, healthy controls (age, gender, skin type), the time of day of the measurement and the environmental conditions. Also, subjects should receive precise instructions before the test, mainly in terms of hygiene procedure or use of topical products. The interpretation of data should not overlook the fact that even small differences in pH may reflect significant modifications at the molecular level. Although it is usually agreed that the pH of the skin surface may influence the cutaneous microflora, much remains to be learnt about the role of the acid mantle of the skin with regard to defensins and other protective mechanisms.
Decrease in Toxic Potential of Mixed Tensides Maintained below the Critical Micelle Concentration: An in vitro Study
Authors: Benassi L. , Bertazzoni G. , Magnoni C. , Rinaldi M. , Fontanesi C. , Seidenari S.
Published: April 2003

Sodium lauryl sulphate (SLS) is an anionic tenside widely utilized in commercial topical preparations that may cause skin irritation. It has been shown that the barrier damage caused by SLS in vivo is lower when SLS is used in combination with other tensides which are able to reduce the critical micelle concentration (CMC). The aim of our study was to evaluate if the cytotoxic effect of SLS is reduced by the association with different tensides also at concentrations well below the CMC. Normal human keratinocytes from plastic surgery were grown in serum-free medium. At subconfluency, the cells were treated with SLS at a dose of 0.0025% in combination with cocamidopropyl betaine, Tween 20 and Tween 80 at the minimum toxic dose. Following tenside treatment, the culture medium was changed, and after 24 h the cells were collected for 3H-thymidine incorporation, the MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide) assay and neutral red (NR) uptake. The cytotoxic effect on normal human keratinocytes, as evaluated by 3H-thymidine incorporation, MTT assay and NR uptake, was significantly decreased by the combination with all the tested tensides. The correlation between cytotoxicity and physical properties was also studied by a conductimetric assay to investigate the mechanism involved in this toxicity reduction.
Skin Penetration of Topical Formulations of Ibuprofen 5%: An in vitro Comparative Study
Authors: Hadgraft J. , Whitefield M. , Rosher P.H.
Published: April 2003

An in vitro isolated human skin technique with known reliable predictive value for in vivo performance was used to compare the skin penetration of the proprietary ibuprofen gel formulation, IbugelTM, with five other commercially available topical formulations containing ibuprofen 5%: IbusprayTM, IbumousseTM, Proflex CreamTM, Fenbid GelTM and Deep Relief GelTM. There was a marked difference between some formulations in the percentage of applied ibuprofen penetrating the skin samples, with IbusprayTM, IbugelTM and IbumousseTM showing the most efficient penetration. The percentage of applied ibuprofen penetrating the skin samples from these formulations was significantly greater (p < 0.05) at all sampling intervals when compared with Proflex CreamTM, Fenbid GelTM or Deep Relief GelTM. By 48 h, the percentage of applied ibuprofen that had penetrated through the skin samples from IbusprayTM, IbugelTM and IbumousseTM was approximately 2.5 times greater than that from Deep Relief GelTM, 3 times greater than that from Proflex CreamTM and 5 times greater than that from Fenbid GelTM. The data demonstrate that, with topically applied preparations, the composition of the vehicle can have a significant impact on the percutaneous penetration of the active medicament. The possible reasons for this are discussed in terms of partition and diffusion phenomena. Different topical presentations of the same drug substance – especially agents like ibuprofen which are intended for subcutaneous action – cannot be assumed to be pharmaceutically and clinically equivalent or indeed interchangeable.
Bioengineering Analysis of Water Hydration: An Overview
Authors: Lee Christine M. , Maibach Howard I.
Published: April 2003

Skin hydration depends on the water-holding capacities of the stratum corneum. One of the techniques used to evaluate skin water content is the water sorption-desorption test. This method involves hydrating the skin with water and then observing the subsequent dehydration activity by means of serial recording with electrical instruments. Currently, three instruments are available to perform such a test: the Skicon® 200, the Corneometer® CM820 and the Nova DPM® 9003. However, each device measures the skin hydration in its own unique parameter, making interexperimental comparisons difficult. Despite this shortcoming, the water sorption-desorption test is a useful tool to measure the dynamic hydration of the skin and the stratum corneum water-holding properties. Pathological skin conditions like atopic dermatitis affect corneum integrity and reduce its ability to hold water. Irritants can also damage the cutaneous barrier and result in decreased hydration. On the contrary, delipidization of the newborn skin with organic solvents increases the water-holding capacity of the stratum corneum. Since the superficial lipid layer covering the skin surface of the newborn limits the sorption of water, its removal allows more water to be absorbed, resulting in a higher water-holding capacity than before. These results have all been determined using the water sorption-desorption test. While the studies produce significant information on skin hydration kinetics, the data are reported in different units. As such, it is difficult to make comparisons between sets of experiments where different electrical instruments are used. Finding a standard parameter unit for measuring skin hydration will benefit dermatological research greatly and allow the results obtained from one instrument to aid in the interpretation of those from another instrument. The high degrees of correlation that exist among the Skicon 200, Corneometer CM820 and Nova DPM 9003 may be the key to future developments of a standardized parameter unit.
Changes of Cutaneous Antioxidant Enzyme Activity in Allergic Contact Dermatitis in Young and Old Individuals
Authors: Bang H.D. , Shin M.H. , Chung J.H. , Cho K.H. , Kim K.H. , Eun H.C.
Published: April 2003

Background: Reactive oxygen species, which are believed to play an important role in the aging process, are also found to be related to allergic skin diseases. Skin reactions to allergens are known to affect the antioxidant defense system such as antioxidant enzymes. Objectives: This study was conducted in order to investigate changes of the enzymatic antioxidant defense system in the lesional skin of allergic contact dermatitis (ACD) in vivoand the influence of skin aging on the changes in the enzymatic antioxidant defense system. Methods: Diphenylcyclopropenone (DPCP) application was used to induce ACD in 10 volunteers under 30 years old and in 10 over 70 years old. Antioxidant enzyme assays were performed in 5 young volunteers and 6 elderly volunteers who showed a weakly positive reaction after DPCP application. The activities of catalase, superoxide dismutase (SOD) and glutathione peroxidase (GPx) were measured in the lesional skin of ACD and in normal control skin. The differences between the two age-distinct groups were also compared and analyzed. Results: Compared with the control site, the DPCP-affected skin revealed a significantly lower catalase activity in both the young and the old groups. Reductions in cutaneous catalase activity were not different in the young and old skins. In contrast, SOD and GPx did not show any significant difference with respect to the presence of contact dermatitis or age. Conclusion: The catalase activity of human skin with ACD decreased in both young and old skins. These findings suggest that cutaneous catalase may play an important role in the pathophysiology of ACD.
Immunologic Effects of Allogeneic versus Autologous Blood Transfusion in Patients Undergoing Radical Oesophagectomy
Authors: Takemura M. , Osugi H. , Takada N. , Kinoshita H. , Higashino M.
Published: April 2003

Blood transfusion is often required in patients undergoing radical oesophagectomy, and is associated with immunosuppression that may worsen postoperative and long-term outcomes. However, the immunologic effects of allogeneic versus autologous transfusion have not been studied in this group of patients. We analyzed 103 patients who underwent radical oesophagectomy for oesophageal cancer, including 45 patients who received allogeneic transfusions (Allo), 16 patients who donated autologous blood but were not transfused (Auto-1) and 42 patients who received autologous transfusions (Auto-2). Peripheral blood lymphocyte subsets and natural killer (NK) cell activity were analyzed for 2 weeks postoperatively. Furthermore, the rate of infectious complications such as pneumonia and wound infection was compared. Patients receiving blood transfusion had decreased CD4+ lymphocyte counts and NK cell activity postoperatively, compared to Auto-1 patients. However, these abnormalities were corrected by day 14 in the Auto-2 group, but not in the Allo group. CD8+ lymphocyte counts were decreased in all groups postoperatively, returning to normal by 14 days in the Auto-1 group only. The rate of infectious complications was significantly higher in the Allo than in the Auto group. Blood transfusion is associated with adverse immunologic effects in patients undergoing radical oesophagectomy. However, autologous blood transfusion is favourable compared to allogeneic transfusion. Autologous transfusion programs should be employed when possible in this group of patients.
Characteristics of Long-Term Live-Donor Renal Allograft Survivors
Authors: El-Agroudy Amgad E. , Bakr Mohamed A. , Hassan Nabil A. , Ismail Amani M. , Ali-El-Dein Bedeir , Shehab El-Dein Ahmed B. , Ghoneim Mohamed A.
Published: April 2003

Background/Aims: Despite the high rate of rejection, allograft failure and patient mortality in the early years of renal allotransplantation, some patients have done-remarkably well. We report here on 62 renal transplant recipients out of 144 patients (43%) who had functioning grafts for more than 15 years (range 15–24 years). Materials: Demographic and follow-up data for patients fulfilling the criteria were reviewed. These patients include 43 males and 19 females, with a mean age at transplantation of 27.5 ± 6.6 years (range 9–43 years), and mean donor age of 30 ± 8.6 years. The donor source was 8 parents, 49 siblings and 5 unrelated. The main causes of end-stage renal disease were chronic pyelonephritis and chronic glomerulonephritis. Twenty-nine patients were treated with cyclosporine (CsA) while 33 patients were primarily immunosuppressed by steroids and azathioprine. Results: Acute rejection episodes occurred in 40 patients (64.3%), out of them 19 patients experienced two or more acute rejection episodes. Univariate analysis showed that recipient and donor age, HLA-DR matching, pre- and post-transplant hypertension, ATN, delayed diuresis and chronic allograft nephropathy are significant risk factors; while recipient age, delayed diuresis and post-transplant hypertension were still significant by multivariate analysis. Conclusions: We concluded that renal transplantation, even in its earliest years and despite the numerous complications, has provided 15 or more years of near-normal life to patients with end-stage renal disease. Certain characteristics of long-term renal allograft survivors include young donor/recipient pairs, good DR matching with less pre- and post-transplantation prevalence of hypertension.
Genetic Evidence That Nitric Oxide Modulates Homocysteine
Authors: Brown Karen S. , Kluijtmans Leo A.J. , Young Ian S. , Woodside Jayne , Yarnell John W.G. , McMaster Dorothy , Murray Liam , Evans Alun E. , Boreham Colin A. , McNulty Helene , Strain J.J. , Mitchell Laura E. , Whitehead Alexander S.
Published: April 2003

Objective—

Mild hyperhomocystenemia is an independent, graded risk factor for cardiovascular disease. Genetic determinants of hyperhomocystenemia include functional polymorphisms in several folate/homocysteine metabolic enzymes. Nitric oxide may also modulate plasma homocysteine (tHcy) concentrations, either by direct inhibition of methionine synthase or via an indirect effect on folate catabolism.

Methods and Results—

The hypothesis that the endothelial nitric oxide synthase (
NOS3
) G894T polymorphism is a genetic determinant of tHcy concentrations was tested in 2 independent healthy adult populations. In both populations,
NOS3
genotype was significantly associated with tHcy concentrations in nonsmokers with low folate (
P
=0.03 for each). Models were constructed to adjust for known determinants of tHcy concentrations and test for interactions between
NOS3
genotype and these determinants in nonsmokers from each population.
NOS3
genotype remained a significant determinant of tHcy concentrations after adjustment. Interactions between
NOS3
genotype and serum folate were significant in both populations, and the interaction between
NOS3
genotype and
MTHFR
C677T genotype was significant in the larger population.

Conclusions—

These data indicate that the
NOS3
894TT genotype is a risk factor for elevated tHcy in healthy nonsmoking adults with low serum folate and supports the hypothesis that nitric oxide modulates homocysteine through an effect on folate catabolism.
Neuropeptide Y–Induced Acceleration of Postangioplasty Occlusion of Rat Carotid Artery
Authors: Li Lijun , Lee Edward W. , Ji Hong , Zukowska Zofia
Published: April 2003

Objective—

Attempts to restore blood flow through atherosclerotic vessels by angioplasty often result in restenosis. Because the role of nerves in this process is unclear, we investigated whether neuropeptide Y (NPY), a sympathetic cotransmitter with vascular mitogenic activities, contributes to postangioplasty restenosis.

Methods and Results—

Carotid artery balloon angioplasty upregulated vascular expression of NPY and its processing enzyme (DPPIV/cd26) and receptors (Y1, Y2, Y5 mRNA and protein) within 6 to 24 hours and stimulated neointima formation and accumulation of NPY in platelets after 14 days. NPY pellets (1 to 10 μg/pellet for 14 days) inserted next to the injured artery elevated platelet and vascular NPY immunoreactivity to stress-like levels and dose-dependently augmented angioplasty-induced neointima. Strikingly, 10 μg NPY for 14 days led to vessel occlusion with an atherosclerotic-like lesion, with thrombus and neointima containing neovessels, macrophages, matrix, and lipids. Y1 or Y5 receptor antagonist completely prevented the effect of NPY and reduced angioplasty-induced neointima by 50%.

Conclusions—

Angioplasty upregulates platelet and vascular NPY systems, which then contribute to neointima formation via Y1 and Y5 receptor activation. Increasing NPY to high stress levels triggers formation of a thrombotic atherosclerotic-like lesion and vessel occlusion. Thus, NPY may be a risk factor for accelerated atherosclerosis, and NPY receptor antagonists may be a possible new treatment for restenosis.
Effect of antiviral treatment in patients with chronic HCV infection and t(14;18) translocation
Authors: Giannelli Francesca , Moscarella Stefania , Giannini Carlo , Caini Patrizio , Monti Monica , Gragnani Laura , Romanelli Roberto Giulio , Solazzo Vera , Laffi Giacomo , La Villa Giorgio , Gentilini Paolo , Zignego Anna Linda
Published: April 2003

Hepatitis C virus (HCV) may be associated with the mixed cryoglobulinemia syndrome and other B-cell lymphoproliferative disorders (LPDs). The t(14;18) translocation may play a pathogenetic role. Limited data are available regarding the effects of antiviral therapy on rearranged B-cell clones. We evaluated the effects of interferon and ribavirin on serum, B-lymphocyte HCV RNA, and t(14; 18) in 30 HCV+, t(14;18)+ patients without either mixed cryoglobulinemia syndrome or other LPDs. The t(14;18) translocation was analyzed by both bcl-2/JH polymerase chain reaction and bcl-2/JH junction sequencing in peripheral blood mononuclear cells in all patients. Fifteen untreated patients with comparable characteristics served as controls. Throughout the study, the presence or absence of both t(14;18) and HCV RNA sequences were, in most cases, associated in the same cell samples. At the end of treatment, t(14;18) was no longer detected in 15 patients (50%) with complete or partial virologic response, whereas it was persistently detected in nonresponders (P < .05), as well as in 14 of 15 control patients. In 4 responder patients, t(14;18) and HCV RNA sequences were no longer detected in blood cells after treatment, but were again detected after viral relapse; the same B-cell clones were involved in the pretreatment and posttreatment periods. In conclusion, this study suggests that antiviral therapy may induce regression of t(14;18)–bearing B-cell clones in HCV+ patients and that this phenomenon may be related, at least in part, to the antiviral effect of therapy. This in turn suggests that antiviral treatment may help prevent or treat HCV-related LPDs.
Telomerase levels control the lifespan of human T lymphocytes
Authors: Röth Alexander , Yssel Hans , Pène Jérôme , Chavez Elizabeth A. , Schertzer Mike , Lansdorp Peter M. , Spits Hergen , Luiten Rosalie M.
Published: April 2003

The loss of telomeric DNA with each cell division contributes to the limited replicative lifespan of human T lymphocytes. Although telomerase is transiently expressed in T lymphocytes upon activation, it is insufficient to confer immortality. We have previously shown that immortalization of human CD8+ T lymphocytes can be achieved by ectopic expression of the human telomerase reverse transcriptase (hTERT) gene, which encodes for the catalytic component of the telomerase complex. To study the role of endogenous hTERT in the lifespan of human T cells, we blocked endogenous hTERT expression by ectopic expression of dominant-negative (DN) hTERT. Cells expressing DN-hTERT had a decreased lifespan and showed cytogenetic abnormalities, including chromosome ends without detectable telomeric DNA as well as chromosome fusions. These results indicate that while endogenous hTERT cannot prevent overall telomere shortening, it has a major influence on the longevity of human T cells. Furthermore, we show that up-regulation of hTERT in T cells upon activation decreases over time in culture. Long-term–cultured T cells also show a decreased expression of c-myc upon activation, resulting in less c-myc–induced transcription of hTERT. Moreover, memory T cells, which have expanded in vivo upon antigen encounter, expressed a lower level of hTERT upon activation than naive cells from the same donor. The observed inverse correlation between telomerase levels and replicative history suggests that telomerase levels in T cells are limiting and increasingly insufficient to sustain their proliferation.
Eotaxin-3 is a natural antagonist for CCR2 and exerts a repulsive effect on human monocytes
Authors: Ogilvie Patricia , Paoletti Samantha , Clark-Lewis Ian , Uguccioni Mariagrazia
Published: April 2003

Eotaxin-3 (CCL26) belongs to the group of CC chemokines that attract eosinophils, basophils, and Th2 lymphocytes. Like eotaxin (CCL11) and eotaxin-2 (CCL24), eotaxin-3 mediates its activity through CCR3. Here we show that eotaxin-3 also binds to CCR2 on monocytes and CCR2-transfected cells. In contrast to monocyte chemotactic protein 1 (MCP-1; CCL2), eotaxin-3 does not trigger intracellular calcium mobilization, enzyme release, or phosphorylation of the mitogen-activated protein (MAP) kinase ERK and induces a weak chemotaxis in monocytes. Instead, eotaxin-3 inhibits MCP-1–mediated responses, thus acting as a natural antagonist for CCR2. This study also demonstrates that eotaxin-3 promotes active movement of monocytes away from a gradient of eotaxin-3 in vitro. This repellent effect is amplified when an additional gradient of MCP-1 is applied, demonstrating that the 2 mechanisms are synergistic. Eotaxin-3 effects on monocytes are largely abolished when cells are pretreated with MCP-1 or CCR2 antagonists. Like MCP-1–mediated migration, repulsion is sensitive to Bordetella pertussis toxin, indicating the involvement of Gi protein–coupled receptors. However, using transfected cells expressing CCR2 we could not detect F-actin formation or an active movement away induced by eotaxin-3, suggesting that either expression of a single receptor type is not sufficient to mediate cell repulsion or that the used transfected cell lines lack additional interaction molecules that are required for reverse migration. Eotaxin-3 was expressed by vascular endothelial cells and was essential for endothelial transmigration of eosinophils. Our data provide a mechanism by which 2 chemokine gradients that are oriented in opposite directions could cooperate in efficiently driving out monocytes from blood vessels into tissue.
The toll-like receptor repertoire of human B lymphocytes: inducible and selective expression of TLR9 and TLR10 in normal and transformed cells
Authors: Bourke Emer , Bosisio Daniela , Golay Josée , Polentarutti Nadia , Mantovani Alberto
Published: April 2003

The present study was designed to investigate the expression of members of the toll-like receptor (TLR) family in human B cells. High-density, resting, and low-density activated tonsillar B cells expressed TLR9 and TLR10 mRNA transcripts at the highest levels. Expression was higher in activated B cells than in resting cells. Analysis of a range of resting and activated human leukocyte populations revealed that mRNA expression of TLR10 was restricted to B cells. Stimulation of resting B cells with anti-μ and anti-CD40 antibodies or with Staphylococcus aureus Cowan I bacteria (SAC) increased expression of TLR9 and TLR10. TLR1 and TLR4 expression were not significantly induced by B-cell activation. Interestingly, a CpG oligonucleotide, a TLR9 agonist, also stimulated TLR9 expression in B cells. Exposure to anti-μ antibodies augmented TLR9 expression, concomitantly and dramatically increasing the responsiveness of B cells to CpG oligonucleotides in terms of proliferation and chemokine (CC chemokine ligand 3 [CCL3] and CCL22) production. Epstein-Barr virus (EBV)–transformed cell lines and other cell lines representative of mature B-cell neoplasias (Burkitt lymphoma, follicular lymphoma, multiple myeloma) expressed TLR9 and/or TLR10, whereas pre-B cell lines were negative. These results show that normal and neoplastic human B lymphocytes express a distinct TLR repertoire including TLR9 and TLR10 and that expression is increased upon engagement of the antigen receptor complex or TLR9 itself. Regulated expression of selected TLRs in B cells is likely to play an important role in linking innate and adaptive immune responses in normal and pathologic conditions.
Risks and outcomes of invasive fungal infections in recipients of allogeneic hematopoietic stem cell transplants after nonmyeloablative conditioning
Authors: Fukuda Takahiro , Boeckh Michael , Carter Rachel A. , Sandmaier Brenda M. , Maris Michael B. , Maloney David G. , Martin Paul J. , Storb Rainer F. , Marr Kieren A.
Published: April 2003

The incidence of invasive mold infections has increased during the 1990s among patients undergoing allogeneic hematopoietic stem cell transplantation (HCT) after myeloablative conditioning. In this study, we determined risk factors for invasive mold infection and mold infection-related death among 163 patients undergoing allogeneic HCT with nonmyeloablative conditioning. The cumulative incidence rates of proven or probable invasive fungal infections, invasive mold infections, invasive aspergillosis, and invasive candidiasis during the first year after allogeneic HCT with nonmyeloablative conditioning were 19%, 15%, 14%, and 5%, respectively, which were similar to those after conventional myeloablative HCT. Invasive mold infections occurred late after nonmyeloablative conditioning (median, day 107), with primary risk factors including severe acute graft-versus-host disease (GVHD), chronic extensive GVHD, and cytomegalovirus (CMV) disease. The 1-year survival after diagnosis of mold infections was 32%. High-dose corticosteroid therapy at diagnosis of mold infection was associated with an increased risk for mold infection–related death. Overall, nonrelapse mortality was estimated at 22% (36 patients) after nonmyeloablative conditioning, of which 39% (14 patients) were mold infection-related (9% of the overall mortality). More effective strategies are needed to prevent invasive mold infections, which currently account for a notable proportion of nonrelapse mortality after nonmyeloablative allogeneic HCT.
Cord blood transplantation provides better reconstitution of hematopoietic reservoir compared with bone marrow transplantation
Authors: Frassoni Francesco , Podestà Marina , Maccario Rita , Giorgiani Giovanna , Rossi Gabriele , Zecca Marco , Bacigalupo Andrea , Piaggio Giovanna , Locatelli Franco
Published: April 2003

Delayed hematopoietic recovery is the main factor precluding a wider use of cord blood (CB) transplants. We hypothesized that this delayed engraftment might not be related to an insufficient number of stem cells in the graft, but to an intrinsic difficulty of these cells to undergo differentiation. To test our hypothesis, 2 groups of children were compared; 12 received a CB transplant and 12 an adult bone marrow (BM) transplant. We studied neutrophil and platelet recovery and, at a median time of approximately 1 year after transplantation, the frequency of colony-forming cells (CFCs) and long-term culture initiating cells (LTC-ICs) in the BM of the 2 groups. Recipients of BM transplants received 1-log more cells and had significantly faster neutrophil and platelet recovery. Conversely, the frequency of committed and early progenitors was significantly higher in the BM of children given CB cells compared with BM transplant recipients (median count of CFC/2 × 104 BM mononuclear cells, 20 versus 11, P = .007; median count of LTC-IC/106 BM mononuclear cells, 8.2 versus 0.2 P = .001). CB, but not adult BM stem cells, can better restore the host hematopoietic progenitor cell reservoir; the delayed engraftment after CB transplantation may reflect the difficulty of CB progenitors to reprogram themselves toward differentiation.
Cardiovascular Disease as a Long-Term Complication of Treatment for Testicular Cancer
Authors: Huddart R.A. , Norman A. , Shahidi M. , Horwich A. , Coward D. , Nicholls J. , Dearnaley D.P.
Published: April 2003

Purpose: To assess the risk of cardiovascular morbidity and cardiac risk factors in long-term survivors of testicular cancer according to treatment received.

Patients and Methods: All resident male patients registered in the United Kingdom between 1982 and 1992 attending for follow-up were eligible for recruitment. Patients completed a current health questionnaire and underwent clinical review, along with hematologic, biochemical, and hormonal profiles. For patients not under routine review, follow-up information was sought from their general practitioner and mortality data were sought from the Office of National Statistics. Descriptive analysis was performed on all variables and comparisons were made among patients treated by orchidectomy and follow-up only, chemotherapy alone (C), radiotherapy alone (RT), and radiotherapy and chemotherapy (C/RT).

Results: Data on cardiovascular events were available on 992 patients. After a median follow-up of 10.2 years, 68 events had been reported, including 18 deaths. After adjusting for age, increased risk for cardiac events was seen after C (relative risk [RR] = 2.59; 95% confidence interval [CI], 1.15 to 5.84; P = .022), RT (RR = 2.40; 95% CI, 1.04 to 5.45; P = .036), and C/RT (RR = 2.78; 95% CI, 1.09 to 7.07; P = .032). There were no significant differences in cardiac risk factors. On multivariate analysis, age, treatment group, free thyroxine, protein, and magnesium levels were associated with cardiovascular disease.

Conclusion: In long-term survivors of testicular cancer, we observed a two-fold or greater risk of developing cardiovascular disease. This was not due to increases in cardiac risk factors, which suggests a direct or indirect treatment effect. These data support the continued research into the minimization of treatment in good-prognosis testicular cancer.
Practical Management of Patients With Chronic Myeloid Leukemia Receiving Imatinib
Authors: Deininger Michael W.N. , O’Brien Stephen G. , Ford John M. , Druker Brian J.
Published: April 2003

The introduction of imatinib, a specific inhibitor of the Bcr-Abl tyrosine kinase, has dramatically changed the management of chronic myeloid leukemia (CML). More than 10,000 patients worldwide have been treated with imatinib in clinical trials, and a large body of information has accumulated about the use of this drug. The purpose of this article is to review practical guidelines in regard to optimal dosing, monitoring, managing common side effects such as myelosuppression, and potential drug interactions. The treatment recommendations are intended to optimize therapy with imatinib while taking into account a patient’s specific circumstances.
Aberrant Regulation of Human Intestinal Proglucagon Gene Expression in the NCI-H716 Cell Line
Authors: Cao Xiemin , Flock Grace , Choi Caroline , Irwin David M. , Drucker Daniel J.
Published: April 2003

Despite interest in understanding glucagon-like peptide-1 (GLP-1) production, the factors important for GLP-1 biosynthesis remain poorly understood. We examined control of human proglucagon gene expression in NCI-H716 cells, a cell line that secretes GLP-1 in a regulated manner. Insulin, phorbol myristate acetate, or forskolin, known regulators of rodent proglucagon gene expression, had no effect, whereas sodium butyrate decreased levels of NCI-H716 proglucagon mRNA transcripts. The inhibitory effect of sodium butyrate was mimicked by trichostatin A but was not detected with sodium acetate or isobutyrate. The actions of butyrate were not diminished by the ERK1/2 inhibitor PD98059, p38 inhibitor SB203580, or soluble guanylate cyclase inhibitor LY83583 or following treatment of cells with KT5823, a selective inhibitor of cGMP-dependent protein kinase. NCI-H716 cells expressed multiple proglucagon gene transcription factors including isl-1, pax-6, pax-2, cdx-2/3, pax-4, hepatocyte nuclear factor (HNF)-3α, HNF-3β, HNF-3γ, and Nkx2.2. Nevertheless, the butyrate-dependent inhibition of proglucagon gene expression was not associated with coordinate changes in transcription factor expression and both the human and rat transfected proglucagon promoters were transcriptionally inactive in NCI-H716 cells. Hence, NCI-H716 cells may not be a physiologically optimal model for studies of human enteroendocrine proglucagon gene transcription.
Estrogen Receptor-α Dependency of Estrogen’s Stimulatory Action on Cancellous Bone Formation in Male Mice
Authors: McDougall Kathleen E. , Perry Mark J. , Gibson Rachel L. , Colley Shane M. , Korach Kenneth S. , Tobias Jonathan H.
Published: April 2003

We examined whether estrogen receptor (ER)α is required for estrogen to stimulate cancellous bone formation in long bones of male mice. 17β-Estradiol (E2) was administered to ERα−/− male mice or wild-type (WT) littermate controls at 40, 400, or 4000 μg/kg by daily sc injection for 28 d and histomorphometric analysis performed at the distal femoral metaphysis. In WT mice, treatment with E2 (40 μg/kg·d) increased the proportion of cancellous bone surfaces undergoing mineralization and stimulated mineral apposition rate. In addition, higher doses of E2 induced the formation of new cancellous bone formation surfaces in WT mice. In contrast, E2 had little effect on any of these parameters in ERα−/− mice. Immunohistochemistry was subsequently performed using an ERα-specific C-terminal polyclonal antibody. In WT mice, ERα was expressed both by cancellous osteoblasts and a significant proportion of mononuclear bone marrow cells. Immunoreactivity was also observed in cancellous osteoblasts of ERα−/− mice, resulting from expression of the activation function-1-deficient 46-kDa ERα isoform previously reported to be expressed in normal osteoblasts and bones of ERα−/− mice. Taken together, our results suggest that estrogen stimulates bone formation in mouse long bones via a mechanism that requires the presence of full-length ERα possessing activation function-1.
Cloning of Transforming Growth Factor-β1 (TGF-β1) and Its Type II Receptor from Zebrafish Ovary and Role of TGF-β1 in Oocyte Maturation
Authors: Kohli Gurneet , Hu Siqin , Clelland Eric , Di Muccio Tamara , Rothenstein Jeffrey , Peng Chun
Published: April 2003

TGF-β is a multifunctional factor involved in regulating a variety of cellular activities. In mammals, TGF-β is known to regulate reproduction, including ovarian functions. The role of TGF-β in lower vertebrates, such as fish, is poorly understood. To examine the role of TGF-β in fish reproduction, cDNAs encoding TGF-β1 and the type II TGF-β receptor (TβRII) were cloned from the zebrafish ovary using PCR- based strategies. The mature peptide region of the zebrafish TGF-β1 shows 70–85% identity with TGF-β1 from other species. The zebrafish TβRII cDNA sequence is the first to be reported from a fish species, and it shows a high level of conservation at the kinase domain. Using RT-PCR, we have detected mRNA expression of TGF-β1, TβRII, as well as its downstream signaling molecules Smad2, 3, and 4 in ovarian follicles at different stages of development. In addition, we have examined the effect of TGF-β1 on oocyte maturation. TGF-β1 significantly inhibited both gonadotropin- and 17α, 20β-dihydroxyprogesterone-induced oocyte maturation in a dose- and time-dependent manner. These findings demonstrate, for the first time, that TGF-β1 plays a role in regulating oocyte maturation in fish and suggest that a TGF-β/Smad signaling pathway is present in the zebrafish ovary.
Abnormal Response of the Neuropeptide Y-Deficient Mouse Reproductive Axis to Food Deprivation But Not Lactation
Authors: Hill Jennifer W. , Levine Jon E.
Published: April 2003

Neuropeptide Y (NPY) plays a key role in both food intake and GnRH secretion. Food deprivation elevates hypothalamic NPY activity and suppresses LH and gonadal steroid secretion. Similarly, lactation up-regulates NPY expression as food consumption increases and estrous cycles cease. These observations suggest that NPY coordinates reproductive suppression in response to energy deficiency; if so, the reproductive axis of NPY knockout (KO) mice should be impervious to lactation and food deprivation. We monitored food consumption, body weight, and estrous cyclicity during lactation in NPY KO mice with large and small litters. NPY KO mice with either litter size resembled wild types (WTs) in weight regulation and food consumption. Large-litter mothers had longer anestrous periods and smaller pups at weaning, but NPY KOs and WTs did not differ in either respect. We also examined the LH response of NPY KO mice to 48 h without food. Basal levels of LH in ovariectomized NPY KO animals decreased in response to fasting, but LH levels in intact and estrogen-treated ovariectomized NPY KO animals did not. In contrast, WTs consistently showed fasting-induced suppression of LH. Our findings suggest that other systems can sustain the hyperphagia of lactation and NPY alone is not responsible for suppressing cyclicity during lactation. Nevertheless, the suppression of basal LH release that accompanies food deprivation in normal female mice appears to require the steroid-dependent actions of NPY.
Role of Signal Transducer and Activator of Transcription 3 in Regulation of Hypothalamic Proopiomelanocortin Gene Expression by Leptin
Authors: Münzberg Heike , Huo Lihong , Nillni Eduardo A. , Hollenberg Anthony N. , Bjørbæk Christian
Published: April 2003

Leptin acts on the brain to regulate body weight and neuroendocrine function. Proopiomelanocortin (POMC) neurons in the hypothalamus are important targets of leptin. These cells express the leptin receptor ObRb, and leptin can regulate POMC mRNA levels, but the cellular mechanisms by which this occurs is unknown. Here we show evidence that leptin stimulates pomc gene transcription via activation of intracellular signal transducer and activator of transcription 3 (STAT3) proteins. In pomc-promoter assays using transfected cells, leptin induces pomc promoter activity. Expression of dominant negative STAT3 strongly suppresses this effect. Furthermore, maximal activation requires the presence of the STAT3-binding site, tyrosine 1138, of ObRb. Mutational analysis identifies a 30-bp promoter element that is required for regulation by leptin. In rats, robust leptin-dependent induction of STAT3 phosphorylation is demonstrated in hypothalamic POMC neurons using double immunohistochemistry. In total, approximately 37% of POMC cells are positive for phospho-STAT3 after leptin treatment. Furthermore, leptin-responsive POMC neurons are concentrated in the rostral region of the hypothalamus. Combined, our data show that a subpopulation of POMC neurons is leptin-responsive and suggest that stimulation of hypothalamic pomc gene expression in these cells requires STAT3 activation. We speculate that STAT3 is critical for leptin-dependent effects on energy homeostasis that are mediated by the central melanocortin system.
Urotensin II is an Autocrine/Paracrine Growth Factor for the Porcine Renal Epithelial Cell Line, LLCPK1
Authors: Matsushita Mika , Shichiri Masayoshi , Fukai Nozomi , Ozawa Naoko , Yoshimoto Takanobu , Takasu Nobuyuki , Hirata Yukio
Published: April 2003

Urotensin-II (UII), a cyclic dodecapeptide with potent cardiovascular effects, has recently been shown to be abundantly expressed in the human kidney and excreted in human urine. To investigate whether UII acts as an autocrine/paracrine growth factor for renal epithelial cells, we have studied the effects of human UII (hUII) on DNA synthesis, cytosolic free Ca2+ concentration ([Ca2+]i), ERK activation, and protooncogene (c-myc) expression in a porcine renal epithelial cell line (LLCPK1). hUII stimulated [3H]thymidine uptake into quiescent cells in a dose-dependent manner (10−9 to 10−7m); this effect was inhibited by a protein kinase C inhibitor (GF109203X), a MAPK kinase inhibitor (PD98059), and a calcium channel blocker (nicardipine). Neither phosphatidyl inositol-3 kinase inhibitors (LY294002, wortmannin) nor p38 kinase inhibitor (SB203580) affected the hUII-induced DNA syntheses. hUII rapidly (within 5 min) and dose-dependently (10−9 to 10−7m) increased [Ca2+]i in fura-2-loaded cells. hUII also caused a rapid and transient activation of ERK1/2 and induction of c-myc. LLCPK1 cells expressed UII mRNA and its receptor GPR14 mRNA, as determined by RT-PCR, and released UII-like immunoreactivity into media. Neutralization of endogenous UII by anti-hUII antibody, but not nonimmune serum, significantly suppressed DNA synthesis. These data suggest that hUII is an autocrine/paracrine growth factor for renal epithelial cells via activation of both protein kinase C and ERK1/2 pathways as well as Ca2+ influx via voltage-dependent Ca2+ channels.
Crystal Structure, Hydrogen Bonding, and 81Br NQR of Low-Temperature Phase of 4-Aminopyridinium Tetrabromoantimonate(III)
Authors: Hashimoto Masao , Terao Hiromitsu , Fuess Hartmut , Svoboda Ingrid , Ehrenberg Helmut
Published: April 2003

The crystal structure of the low-temperature phase (LTP) of the title compound was determined at 220 K (monoclinic, P21⁄c). The 4-aminopyridinium cations (4-NH2C5H4NH+) were found to be ordered in LTP, while being severely disordered in the room-temperature phase (monoclinic, C2⁄c). The tetrabromoantimonate anions (SbBr4−) were incorporated into the infinite polyanion chains of irregular SbBr6 octahedra with two-edges sharing. The trans-Br–Sb–Br moiety in the SbBr4− anion was approximately symmetric differing from the asymmetric Br–Sb···Br moiety found in LTP of pyridinium tetrabromoantimonate(III). The N–H moieties in both of the pyridine ring and the amino (–NH2) group participate in the formation of N–H···Br hydrogen bonds. It was shown that the 81Br NQR spectrum of LTP is closely related to the anion structure and the hydrogen bonds. The distinctive anion structures, as well as the different types of phase transitions, exhibited by the PyH and 4-APyH salts, seemed to be attributable to the difference in the hydrogen bond scheme between them.
Dynamic Quantification of the Atom Rearrangement Behavior with Al-Mg 5052 Undercooling Liquid Film
Authors: Tokuchi Eisaku , Asai Kimioku
Published: April 2003

The atom rearrangement behaviors along with the rapid solidification and high temperature oxidation of an interdendritic liquid (liquid film) with an arc spot-fused A5052 sheet were experimentally investigated in comprehensive terms of the solidification progress, concentration, and temperature by using high-speed breaking and breakage techniques. The latter is a new quenching method with an aqueous solution of surface-active agents to prevent the fractured section from the selective diffusion of Mg coupled with high temperature oxidation. Continuous liquid films of the fractured-section (solidification) layers were analyzed by XPS as well as fractography with FE-SEM . In a surfactant experiment, the results verified that the characteristic diffusion during oxidation occurred with the driving force being electrostatic attraction. Concerning the solidification behavior of rapidly solidifying columnar dendrite, the results of XPS measurements indicated that after a tip undercooling of about 25 °C the liquid solute concentration increased exponentially with decreasing temperature, while taking a solute concentration path below the T0 curve (in binary system analysis). The fractography was in qualitative agreement with the solute concentration result in terms of the mass balance by showing restricted lateral solidification progress.