bluuebunny/crossref_metadata_2025 huggingface dataset:
Enhanced sodium-dependent extrusion of magnesium in mutant cells established from a mouse renal tubular cell line
Authors: Watanabe Masaru , Konishi Masato , Ohkido Ichiro , Matsufuji Senya
Published: May 2005

To study the regulatory mechanisms of intracellular Mg2+ concentration ([Mg2+]i) in renal tubular cells as well as in other cell types, we established a mutant strain of mouse renal cortical tubular cells that can grow in culture media with very high extracellular Mg2+ concentrations ([Mg2+]o > 100 mM: 101Mg-tolerant cells). [Mg2+]i was measured with a fluorescent indicator furaptra (mag-fura 2) in wild-type and 101Mg-tolerant cells. The average level of [Mg2+]i in the 101Mg-tolerant cells was kept lower than that in the wild-type cells either at 51 mM or 1 mM [Mg2+]o. When [Mg2+]o was lowered from 51 to 1 mM, the decrease in [Mg2+]i was significantly faster in the 101Mg-tolerant cells than in the wild-type cells. These differences between the 101Mg-tolerant cells and the wild-type cells were abolished in the absence of extracellular Na+ or in the presence of imipramine, a known inhibitor of Na+/Mg2+ exchange. We conclude that Na+-dependent Mg2+ transport activity is enhanced in the 101Mg-tolerant cells. The enhanced Mg2+ extrusion may prevent [Mg2+]i increase to higher levels and may be responsible for the Mg2+ tolerance.
PPARγ agonists exert antifibrotic effects in renal tubular cells exposed to high glucose
Authors: Panchapakesan U. , Sumual S. , Pollock C. A. , Chen X.
Published: May 2005

Peroxisome proliferator-activated receptor-γ (PPARγ) are ligand-activated transcription factors that regulate cell growth, inflammation, lipid metabolism, and insulin sensitivity. We recently demonstrated that PPARγ agonists limit high glucose-induced inflammation in a model of proximal tubular cells (PTC; Panchapakesan U, Pollock CA, and Chen XM. Am J Physiol Renal Physiol 287: F528–F534, 2004). However, the role of PPARγ in the excess extracellular matrix production is largely unknown. We evaluated the effect of 24- to 48-h 8 μM l-805645 or 10 μM pioglitazone on 25 mM d-glucose-induced markers of fibrosis in HK-2 cells. High d-glucose induced nuclear binding of activator protein-1 (AP-1) to 140.8 ± 10.9% ( P < 0.05), which was attenuated with L-805645 and pioglitazone to 82.3 ± 14.4 ( P < 0.01 vs. high d-glucose) and 99.3 ± 12.2% ( P < 0.05 vs. high d-glucose), respectively. High d-glucose increased total production of transforming growth factor (TGF)-β1139.6 ± 6.5% ( P < 0.05), which was reversed with L-805645 and pioglitazone to 68.73 ± 5.7 ( P < 0.01 vs. high d-glucose) and 112 ± 13.6% ( P < 0.05 vs. high d-glucose). L-805645 and pioglitazone reduced high d-glucose-induced fibronectin from 156.0 ± 24.9 ( P < 0.05) to 81.9 ± 16.0 and 57.4 ± 12.7%, respectively (both P < 0.01 vs. high d-glucose). Collagen IV was not induced by high d-glucose. L-805645 and pioglitazone suppressed collagen IV to 68.0 ± 14.5 ( P < 0.05) and 46.5 ± 11.6% ( P < 0.01) vs. high d-glucose, respectively. High d-glucose increased the nuclear binding of NF-κB to 167 ± 22.4% ( P < 0.05), which was not modified with PPARγ agonists. In conclusion, PPARγ agonists exert antifibrotic effects in human PTC in high glucose by attenuating the increase in AP-1, TGF-β1, and the downstream production of the extracellular matrix protein fibronectin.
Kallikrein gene transfer reduces renal fibrosis, hypertrophy, and proliferation in DOCA-salt hypertensive rats
Authors: Xia Chun-Fang , Bledsoe Grant , Chao Lee , Chao Julie
Published: May 2005

In DOCA-salt hypertension, renal kallikrein levels are increased and may play a protective role in renal injury. We investigated the effect of enhanced kallikrein levels on kidney remodeling of DOCA-salt hypertensive rats by systemic delivery of adenovirus containing human tissue kallikrein gene. Recombinant human kallikrein was detected in the urine and serum of rats after gene delivery. Kallikrein gene transfer significantly decreased DOCA- and salt-induced proteinuria, glomerular sclerosis, tubular dilatation, and luminal protein casts. Sirius red staining showed that kallikrein gene transfer reduced renal fibrosis, which was confirmed by decreased collagen I and fibronectin levels. Furthermore, kallikrein gene delivery diminished myofibroblast accumulation in the interstitium of the cortex and medulla, as well as transforming growth factor (TGF)-β1 immunostaining in glomeruli. Western blot analysis and ELISA verified the decrease in immunoreactive TGF-β1 levels. Kallikrein gene transfer also significantly reduced kidney weight, glomerular size, proliferating tubular epithelial cells, and macrophages/monocytes. Reduction of proliferation and hypertrophy was associated with reduced levels of the cyclin-dependent kinase inhibitor p27Kip1, and the phosphorylation of c-Jun NH2-terminal kinase (JNK) and extracellular signal-regulated kinase (ERK). The protective effects of kallikrein were accompanied by increased urinary nitrate/nitrite and cGMP levels, and suppression of superoxide formation. These results indicate that kallikrein protects against mineralocorticoid-induced renal fibrosis glomerular hypertrophy, and renal cell proliferation via inhibition of oxidative stress, JNK/ERK activation, and p27Kip1and TGF-β1 expression.
Adapting Locomotion to Different Surface Compliances: Neuromuscular Responses and Changes in Movement Dynamics
Authors: Marigold Daniel S. , Patla Aftab E.
Published: May 2005

Knowledge of how the nervous system deals with surfaces with different physical properties such as compliance that challenge balance during locomotion is of importance as we are constantly faced with these situations every day. The purpose of this study was to examine the control of center of mass (COM) and lower limb dynamics and recovery response modulation of muscle activity during locomotion across an unexpected compliant surface and in particular, scaling behavior across different levels of compliance. Eight young adults walked along a walkway and stepped on an unexpected compliant surface in the middle of the travel path. There were three different levels of surface compliance, and participants experienced either no compliant surface or one of the three compliant surfaces during each trial that were presented in a blocked or random fashion. Whole body kinematics were collected along with surface electromyography (EMG) of selected bilateral lower limb and trunk muscles. The recovery response to the first compliant-surface trial demonstrated muscle onset latencies between 97 and 175 ms, and activity was modulated while on the compliant surface. Vertical COM trajectory was not preserved after contact with the compliant surface: peak vertical COM, while on the compliant surface was lower than when on stable ground. Perturbed-limb knee flexion after toe-off increased with increased surface compliance, which enabled toe clearance with the ground to be similar to control trials. The results suggest that stepping off of a compliant surface is actively modulated by the CNS and is geared toward maintaining dynamic stability.
Inhibition of Perforant Path Input to the CA1 Region by Serotonin and Noradrenaline
Authors: Otmakhova Nonna A. , Lewey Jennifer , Asrican Brent , Lisman John E.
Published: May 2005

Bath-applied monoamines—dopamine (DA), serotonin (5-HT), and noradrenaline (NE)—strongly suppress the perforant path (PP) input to CA1 hippocampal region with very little effect on the Schaffer collaterals (SC) input. The effect of DA action on PP field excitatory postsynaptic potential (fEPSP) has been characterized in detail, but relatively little is known about the NE and 5-HT effects. Here we show that the maximal inhibition of the PP fEPSP by NE is ∼55%, whereas 5-HT inhibition is weaker (∼35%). The half-maximal inhibitory concentration of both 5-HT and NE is ∼1 μM. Neither NE nor 5-HT affected paired-pulse facilitation, suggesting that the effect is not presynaptic. This is in contrast to DA, which does have a presynaptic effect. The NE effect was blocked by α2 antagonists, whereas the α1 antagonist corynanthine and β-antagonist propranolol were ineffective. The effect of 5-HT was mimicked by the agonist, 5-carboxamidotryptamine maleate (5-CT), and not affected by adrenergic and dopaminergic antagonists. To determine the 5-HT receptors involved, we tested a number of 5-HT antagonists, but none produced a complete suppression of the 5-HT effect. Of these, only the 5-HT7 and 5-HT2 antagonists produced weak but significant inhibition of 5-HT effect. We conclude that NE inhibits the PP fEPSP through postsynaptic action on α2-adrenoceptors and that 5-HT7, 5-HT2, and some other receptor may be involved in 5-HT action in PP.
Postnatal Development Differentially Affects Voltage-Activated Calcium Currents in Respiratory Rhythmic Versus Nonrhythmic Neurons of the Pre-Bötzinger Complex
Authors: Elsen Frank P. , Ramirez Jan-Marino
Published: May 2005

The mammalian respiratory network reorganizes during early postnatal life. We characterized the postnatal developmental changes of calcium currents in neurons of the pre-Bötzinger complex (pBC), the presumed site for respiratory rhythm generation. The pBC contains not only respiratory rhythmic (R) but also nonrhythmic neurons (nR). Both types of neurons express low- and high-voltage-activated (LVA and HVA) calcium currents. This raises the interesting issue: do calcium currents of the two co-localized neuron types have similar developmental profiles? To address this issue, we used the whole cell patch-clamp technique to compare in transverse slices of mice LVA and HVA calcium current amplitudes of the two neuron populations (R and nR) during the first and second postnatal week (P0–P16). The amplitude of HVA currents did not significantly change in R pBC-neurons (P0–P16), but it significantly increased in nR pBC-neurons during P8–P16. The dehydropyridine (DHP)-sensitive current amplitudes did not significantly change during the early postnatal development, suggesting that the observed amplitude changes in nR pBC-neurons are caused by (DHP) insensitive calcium currents. The ratio between HVA calcium current amplitudes dramatically changed during early postnatal development: At P0–P3, current amplitudes were significantly larger in R pBC-neurons, whereas at P8–P16, current amplitudes were significantly larger in nR pBC-neurons. Our results suggest that calcium currents in pBC neurons are differentially altered during postnatal development and that R pBC-neurons have fully expressed calcium currents early during postnatal development. This may be critical for stable respiratory rhythm generation in the underlying rhythm generating network.
Motor Cortex Neural Correlates of Output Kinematics and Kinetics During Isometric-Force and Arm-Reaching Tasks
Authors: Sergio Lauren E. , Hamel-Pâquet Catherine , Kalaska John F.
Published: May 2005

We recorded the activity of 132 proximal-arm-related neurons in caudal primary motor cortex (M1) of two monkeys while they generated either isometric forces against a rigid handle or arm movements with a heavy movable handle, in the same eight directions in a horizontal plane. The isometric forces increased in monotonic fashion in the direction of the force target. The forces exerted against the handle in the movement task were more complex, including an initial accelerating force in the direction of movement followed by a transient decelerating force opposite to the direction of movement as the hand approached the target. EMG activity of proximal-arm muscles reflected the difference in task dynamics, showing directional ramplike activity changes in the isometric task and reciprocally tuned “triphasic” patterns in the movement task. The apparent instantaneous directionality of muscle activity, when expressed in hand-centered spatial coordinates, remained relatively stable during the isometric ramps but often showed a large transient shift during deceleration of the arm movements. Single-neuron and population-level activity in M1 showed similar task-dependent changes in temporal pattern and instantaneous directionality. The momentary dissociation of the directionality of neuronal discharge and movement kinematics during deceleration indicated that the activity of many arm-related M1 neurons is not coupled only to the direction and speed of hand motion. These results also demonstrate that population-level signals reflecting the dynamics of motor tasks and of interactions with objects in the environment are available in caudal M1. This task-dynamics signal could greatly enhance the performance capabilities of neuroprosthetic controllers.
Identity Deception: Not a Crime for a Stem Cell
Authors: Leri Annarosa , Kajstura Jan , Anversa Piero
Published: May 2005

Stem cell transdifferentiation in the adult organism is the most common and questioned mechanism of growth and repair. Recent data suggest that adult stem cells are capable of generating mature cells beyond their own tissue boundaries, a process called developmental plasticity. To date, the most versatile cell discovered is the bone marrow progenitor cell.
Intracellular Signaling: Spatial and Temporal Control
Authors: Moraru Ion I. , Loew Leslie M.
Published: May 2005

Cells integrate many inputs through complex networks of interacting signaling pathways. Systems approaches as well as computer-aided reductionist approaches attempt to “untangle the wires” and gain an intimate understanding of cells. But “understanding” any system is just the way that the human mind gains the ability to predict behavior. Computer simulations are an alternative way to achieve this goal—quite possibly the only way for complex systems. We have new tools to probe large sets of unknown interactions, and we have amassed enough detailed information to quantitatively describe many functional modules. Cell physiology has passed the threshold: the time to begin modeling is now.
Uncoupling protein 1 in fish uncovers an ancient evolutionary history of mammalian nonshivering thermogenesis
Authors: Jastroch Martin , Wuertz Sven , Kloas Werner , Klingenspor Martin
Published: May 2005

Uncoupling proteins (UCPs) increase proton leakage across the inner mitochondrial membrane. Thereby, UCP1 in brown adipose tissue dissipates proton motive force as heat. This mechanism of nonshivering thermogenesis is considered as a monophyletic trait of endothermic placental mammals that emerged about 140 million years ago and provided a crucial advantage for life in the cold. The paralogues UCP2 and UCP3 are probably not thermogenic proteins but convey mild uncoupling, which may serve to reduce the rate of mitochondrial reactive oxygen species production. Both are present in endotherms (mammals and birds), but so far only UCP2 has been identified in ectothermic vertebrates (fish and amphibia). The evolution of UCPs is of general interest in the search for the origin of mammalian UCP1-mediated nonshivering thermogenesis. We here show the presence of UCP1 and UCP3 in ectothermic teleost fish species using comparative genomics, phylogenetic inference, and gene expression analysis. In the common carp ( Cyprinus carpio), UCP1 is predominantly expressed in the liver and strongly diminished in response to cold exposure, thus contrasting the cold-induced expression of mammalian UCP1 in brown adipose tissue. UCP3 mRNA is only found in carp skeletal muscle with expression levels increased fivefold in response to fasting. Our findings disprove the monophyletic nature of UCP1 in placental mammals and demonstrate that all three members of the core UCP family were already present before the divergence of ray-finned and lobe-finned vertebrate lineages about 420 million years ago.
Muscles of mice deficient in α-sarcoglycan maintain large masses and near control force values throughout the life span
Authors: Consolino Christina M. , Duclos Franck , Lee Jane , Williamson Roger A. , Campbell Kevin P. , Brooks Susan V.
Published: May 2005

α-Sarcoglycan-deficient ( Sgca-null) mice provide potential for elucidating the pathogenesis of limb girdle muscular dystrophy type 2D (LGMD 2D) as well as for studying the effectiveness of therapeutic strategies. Skeletal muscles of Sgca-null mice demonstrate an early onset of extensive fiber necrosis, degeneration, and regeneration, but the progression of the pathology and the effects on muscle structure and function throughout the life span are not known. Thus the phenotypic accuracy of the Sgca-null mouse as a model of LGMD 2D has not been fully established. To investigate skeletal muscle structure and function in the absence of α-sarcoglycan throughout the life span, we analyzed extensor digitorum longus and soleus muscles of male and female Sgca-null and wild-type mice at 3, 6, 12, and 18 mo of age. Maximum isometric forces and powers were measured in vitro at 25°C. Also determined were individual myofiber cross-sectional areas and numbers, water content, and the proportion of the cross section occupied by connective tissue. Muscle masses were 40–100% larger for Sgca-null compared with age- and gender-matched wild-type mice, with the majority of the increased muscle mass for Sgca-null mice attributable to greater connective tissue and water contents. Although the greater mass of muscles in Sgca-null mice was primarily noncontractile material, absolute forces and powers were maintained near control levels at all ages, indicating a successful adaptation to the deficiency in α-sarcoglycan not observed at any age in LGMD 2D patients.
Glucocorticoids for Human Skin: New Aspects of the Mechanism of Action
Authors: Schäfer-Korting M. , Kleuser B. , Ahmed M. , Höltje H.-D. , Korting H.C.
Published: May 2005

Topical glucocorticoids have always been considered first-line drugs for inflammatory diseases of the skin and bronchial system. Applied systemically, glucocorticoids are used for severe inflammatory and immunological diseases and the inhibition of transplant rejection. Owing to the progress in molecular pharmacology, the knowledge of the mechanism of action has increased during the last years. Besides distinct genomic targets, which are due to the activation of specific cytoplasmatic receptors resulting in the (trans-) activation or (trans-) repression of target genes, there are non-genomic effects on the basis of the interference with membrane-associated receptors as well as with membrane lipids. In fact, various glucocorticoids appear to differ with respect to the relative influence on these targets. Thus, the extended knowledge of glucocorticoid-induced cellular signalling should allow the design and development of even more specifically acting drugs – as it has been obtained with other steroids, e.g. estrogens for osteoporosis prevention.
Characterization of Large-Conductance Ca2+-Dependent and -Independent K+ Channels in HaCaT Keratinocytes
Authors: Ivanchenko E. , Markwardt F.
Published: May 2005

To characterize ion channels expressed in cell membrane of human keratinocytes, patch-clamp recordings were carried out in HaCaT cells. Two types of large-conductance K+ channels (about 250 pS) were measured. One type was activated by micromolar concentrations of intracellular Ca2+ ions ([Ca2+]i) and membrane depolarization, the other was [Ca2+]i independent. The channels were neither dependent on intracellular ATP nor Mg2+ nor on membrane stretch. We conclude that HaCaT keratinocytes express Ca2+-dependent maxi K+ channels and still unknown large Ca2+-independent K+ channels. These K+ channels may affect the proliferation and differentiation of human keratinocytes by the influence on the resting potential, which may control the Ca2+ influx across the cell membrane.
Antagonism of Epidermal Growth Factor Receptor Tyrosine Kinase Ameliorates the Psoriatic Phenotype in Organ-Cultured Skin
Authors: Varani J. , Lateef H. , Fay K. , Elder J.T.
Published: May 2005

Psoriatic plaque skin incubated for eight days in organ culture in the presence of a potent epidermal growth factor (EGF) receptor tyrosine kinase (RTK) antagonist reverted to a more normal histological appearance, while untreated psoriatic plaque skin retained histological features associated with the psoriatic phenotype. In concomitant studies it was shown that the EGF-RTK antagonist had no significant effect on histological features of non-psoriatic skin and no effect on dermal function, i.e. elaboration of both type I procollagen and matrix metalloproteinase-1 (MMP-1; interstitial collagenase). When human epidermal keratinocytes were treated with the EGF-RTK antagonist in monolayer culture, growth inhibition was seen (ED50 = approximately 0.06 µM). When dermal fibroblasts were exposed to the EGF-RTK antagonist in monolayer culture, proliferation, MMP-1 and type I procollagen production were essentially unaffected at concentrations which interfered with keratinocyte growth (up to 1 µM). The capacity of the EGF-RTK antagonist to modulate the histological features of psoriatic skin in organ culture under conditions in which normal skin architecture and dermal function are largely unaffected suggests a potential for anti-psoriatic therapy.
Transepidermal Water Loss Dynamics of Human Vulvar and Thigh Skin
Authors: Warren R. , Bauer A. , Greif C. , Wigger-Alberti W. , Jones M.B. , Roddy M.T. , Seymour J.L. , Hansmann M.A. , Elsner P.
Published: May 2005

Refinement in procedures to assess skin surface water loss (SSWL) dynamics of the vulvar skin on a large sample of subjects (60) is described and compared to another semi-occluded skin site, the inner thigh. Vulvar SSWL significantly decreased over a 30-min period from 46.2 ± 2.6 (SE) to 24.7 ± 1.6 g m–2 h (p < 0.001). The inner thigh, another semi-occluded region, showed no similar pattern for SSWL (6.2 ± 0.3 to 6.6 ± 0.5 g m–2 h), and the values were significantly less than those for vulvar skin. There was no significant effect of age, body mass index or atopic status on vulvar SSWL.
Phase I and Pharmacokinetic Evaluation of Intravenous Hyaluronic Acid in Combination with Doxorubicin or 5-Fluorouracil
Authors: Rosenthal Mark A. , Gibbs Peter , Brown Tracey J. , Wong Shirley , Uren Shannon , Ellis Andrew , Li Lingli , Heldin Paraskevi , Fox Richard M.
Published: May 2005

Background: Pre-clinically, hyaluronan (HA) has been demonstrated to systemically target chemotherapeutic drugs to tumours while ameliorating treatment toxicities. This study is a preliminary clinical investigation to determine if HA could be safely used in combination with 5-fluorouracil (5-FU) and doxorubicin (DOX). Methods: Thirty patients with metastatic cancer were intravenously administered 500 mg/m2 HA in combination with escalating doses of DOX (30–60 mg/m2) or 5-FU (cumulative dose of 1,350–2,250 mg/m2 per cycle). The effect of pre-administration of 20 mg/m2 of folinic acid on HA/5-FU chemotherapy was also investigated. Patients were randomized to receive either HA/chemotherapy or chemotherapy alone in their first treatment cycle and vice versa for the second cycle. Patients received HA and chemotherapy in all subsequent cycles. Results:Treatment was well tolerated, tumour responses were observed and the co-administration of HA did not alter the pharmacokinetics of clinically relevant doses of 5-FU or DOX. Conclusion:High doses of intravenous high-molecular-weight HA can be safely co-administered with clinical doses of chemotherapy without significantly altering the toxicity or pharmacokinetics of the drugs or HA.
Fosfomycin Tromethamine in Uncomplicated Urinary Tract Infections: A Clinical Study
Authors: Bonfiglio Giovanni , Mattina Roberto , Lanzafame Antonina , Cammarata Esmeralda , Tempera Gianna
Published: May 2005

The aim of our study was to verify if the empiric therapy with a single dose of 3 g fosfomycin tromethamine in patients with uncomplicated urinary tract infections (UTIs) was able to clinically resolveand to microbiologically eradicate the infection. A total of 387 out of the 400 patients (274 cases with acute and 113 cases with recurrent uncomplicated UTIs) were enrolled in the clinical study. Clinical and microbiological assessments were performed before and at 8–10 days after the administration. At follow-up high clinical recovery (88.9%) and bacteriological (94.9%) eradication rates were achieved. Gastrointestinal side effects were found in only 4.3% of patients. In conclusion, a single-dose administration regimen of fosfomycin tromethamine should be encouraged as a first choice of drug therapy for uncomplicated UTIs.
Exacerbation of Psoriasis due to Peginterferon alpha-2b plus Ribavirin Treatment of Chronic Active Hepatitis C
Authors: Kartal Elif Doyuk , Colak Hasan , Özgünes Ilhan , Usluer Gaye
Published: May 2005

A 45-year-old woman with chronic hepatitis C was treated with peginterferon alpha-2b in combination with ribavirin, which activated psoriasis. In the patient, psoriasis reoccurred after being inactive for 10 years. One week after institution of therapy, she presented with plaques of psoriasis at her injection site having been administered a first dose of peginterferon alpha-2b. Psoriatic lesions of the patient worsened dramatically during therapy. Cutaneous lesions appeared at the injection sites, in the face, the scalp hair, at the back of the ears, under the breasts and armpits, and in the regions of the anus and elbows. In the second week of treatment, transaminase levels of the patient regressed to normal. Detectable hepatitis C virus RNA in the serum was eliminated at week 14 of treatment. Because of the severity of psoriatic disease activity, therapy was discontinued at the end of 14 weeks after treatment onset. This side effect of peginterferon alpha-2b plus ribavirin treatment should be kept in mind in the treatment of patients with a history of psoriasis combined with active hepatitis C.
Synergic Antinociceptive Interaction between Tramadol and Gabapentin after Local, Spinal and Systemic Administration
Authors: Granados-Soto Vinicio , Argüelles Carlos F.
Published: May 2005

The possible interaction between tramadol and gabapentin on formalin-induced nociception in the rat was assessed. Tramadol, gabapentin or a fixed-dose ratio combination of gabapentin and tramadol were administered peripherally, spinally and orally to rats, and the antinociceptive effect was determined in the 1% formalin test. Isobolographic analyses were used to define the nature of the interactions between drugs. Tramadol, gabapentin and tramadol-gabapentin combinations produced a dose-dependent antinociceptive effect when administered locally, spinally or orally. ED30 values were estimated for the individual drugs and isobolograms were constructed. Theoretical ED30 values for the combination estimated from the isobolograms were 126.8 ± 11.1 µg/paw, 23.1 ± 2.6 µg/rat, and 2.23 ± 0.32 mg/kg for the local, intrathecal and oral routes, respectively. These values were significantly higher than the actually observed ED30 values which were 13.3 ± 2.1 µg/paw, 8.1 ± 0.6 µg/rat and 0.71 ± 0.10 mg/kg, indicating a synergistic interaction. Although efficacy was not improved, local peripheral administration resulted in the highest increase in potency, being about tenfold. Spinal and systemic administration increased potency threefold. Data indicate that low doses of the tramadol-gabapentin combination can interact synergistically to reverse formalin-induced nociception and may represent a therapeutic advantage for clinical treatment of inflammatory pain.
Papillary Microcarcinomas of the Thyroid with Preoperatively Detectable Lymph Node Metastasis Show Significantly Higher Aggressive Characteristics on Immunohistochemical Examination
Authors: Ito Yasuhiro , Uruno Takashi , Takamura Yuuki , Miya Akihiro , Kobayashi Kaoru , Matsuzuka Fumio , Kuma Kanji , Miyauchi Akira
Published: May 2005

Objectives: We recently demonstrated that papillary microcarcinomas with preoperatively detectable node metastasis in the lateral compartment on ultrasonography (clinically apparent metastasis) show worse postoperative relapse-free survival than those with no metastasis or metastasis that could not be detected preoperatively, but was confirmed by pathological examination after surgery (occult metastasis). In this study, we investigated difference in the aggressive characteristics of microcarcinoma of this type from various perspectives. Materials and Methods: We immunohistochemically examined the expression of cell proliferating markers, Ki-67, cyclin D1, p27, and retinoblastoma gene product (pRb), apoptotic markers, single-strand DNA (ssDNA), and metastatic suppressor, kangai-1 (KAI-1) for 19 microcarcinoma patients with clinically apparent metastasis, 14 patents with occult metastasis, and 22 patients without metastasis. Results: Cases of clinically apparent metastasis showed increased cyclin D1 expression together with decreased p27 expression and higher levels of pRb and Ki-67 expression. Furthermore, ssDNA expression was higher and bcl-2 expression was lower in these cases, while KAI-1 expression was significantly reduced. There was no significant difference in the expression of these proteins between cases demonstrating no and occult metastases. Conclusion: These findings suggest that cases of clinically apparent metastasis show significantly higher growth based on cell proliferating activity, apoptosis, and expression of metastatic suppressor than those demonstrating no or occult metastases.
Syndecan-1 Expression – A Novel Prognostic Marker in Pancreatic Cancer
Authors: Juuti Anne , Nordling Stig , Lundin Johan , Louhimo Johanna , Haglund Caj
Published: May 2005

Objective: Syndecan-1 is a transmembrane receptor that participates in cell-cell and cell-matrix interactions, cell proliferation and cell migration. Expression of syndecan-1 is downregulated in many cancers, but in pancreatic ductal adenocarcinoma it is upregulated. Method: We studied the immunohistochemical expression of syndecan-1 in 144 pancreatic adenocarcinomas and evaluated the prognostic value of syndecan-1 expression. Formalin-fixed, paraffin-embedded specimens were stained with mouse monoclonal antibody B-B4 against human syndecan-1. The epithelial and stromal staining was separately evaluated and compared with patient survival, clinical stage and histological grade. Result: Epithelial immunoreactivity was observed in most of the pancreatic carcinoma samples: in 70 (49%) of the samples the epithelial staining was weak, in 48 (33%) moderate, in 18 (12%)strong and in only 8 (6%) of the samples the epithelial staining was negative. Stromal staining was weak in 24 (17%), moderate in 31 (22%), strong in 11 (8%) and negative in 78 (54%) of the pancreatic carcinoma samples. Lack of stromal expression predicted a better prognosis (p = 0.002; HR 1.7) and it was independent of stage (p = 0.01; HR = 1.5) and grade (p = 0.0004; HR 2.1) in multivariate analysis. Epithelial expression predicted better prognosis for patients that underwent surgery for cure (n = 94, p = 0.03). Conclusion: Stromal syndecan-1 expression is an independent prognostic marker in pancreatic cancer, whereas epithelial syndecan-1 expression predicts better prognosis only in resectable disease.
Is There an Indication for High-Dose Chemotherapy in the Treatment of Bone and Soft-Tissue Sarcoma?
Authors: Kasper Bernd , Ho Anthony D. , Egerer Gerlinde
Published: May 2005

Sarcomas represent a rare and heterogeneous disease, and the prognosis of patients with unresectable or advanced metastatic bone and especially soft-tissue sarcomas remains poor, with a disease-free survival of less than 10% at 5 years. Only few chemotherapeutic agents have been identified to be active, with reported response rates for doxorubicin, epirubicin and ifosfamide above 20%. Although combination therapy with different chemotherapeutic substances results in higher response rates, superiority against single-agent chemotherapy in terms of survival has not been demonstrated yet. Regarding anthracyclines, and ifosfamide in particular, a dose-response relationship has been shown, and high-dose chemotherapy with stem cell support has also been evaluated by several investigators. However, all studies comprised small patient cohorts and included very heterogeneous histological subtypes of soft-tissue sarcoma. Moreover, randomized trials are missing. Nevertheless, higher doses of chemotherapy result in higher response rates possibly correlating with longer survival. Finally, well-designed randomized trials should be performed, preferably in younger patients and in the context of an interdisciplinary treatment approach. In this review, we provide an overview of the literature concerning high-dose chemotherapy with hematopoietic stem cell support in the treatment of bone and soft-tissue sarcomas. On the basis of our own data, we would like to emphasize the importance of high-dose chemotherapy in the treatment of sarcomas.
Secretion of Parathyroid Hormone Oscillates Depending on the Change in Serum Ionized Calcium during Hemodialysis and May Affect Bone Metabolism
Authors: Kitahara Tokuyuki , Ueki Kazue , Kuroiwa Takashi , Kaneko Yoriaki , Hiromura Keiju , Nojima Yoshihisa
Published: May 2005

Background: The current study was undertaken to clarify the dynamic response of parathyroid hormone (PTH) during hemodialysis and to determine whether or not such dynamic change of PTH affects bone turnover. Methods: Serum ionized calcium (iCa) and intact PTH (iPTH) were measured in 66 dialysis patients before (basal) and after each hemodialysis. The changes of iCa (ΔiCa) and iPTH (ΔiPTH) were defined as [postdialysis iCa – basal iCa] mM, and [(postdialysis iPTH – basal iPTH)/basal iPTH] × 100%, respectively. We also investigated the data of the patients divided into four groups based on their basal iPTH levels; group 1 (iPTH <60 pg/ml, n = 17), 2 (≧60 to <150, n = 20), 3 (≧150 to <300, n = 15), and 4 (≧300, n = 14). Results: While iCa and iPTH changed variably during each hemodialysis procedure, there was a highly significant inverse correlation between ΔiCa and ΔiPTH (r = –0.761, p < 0.0001). Regression coefficients between ΔiPTH and ΔiCa were –519.1, –311.2, –268.1, and –194.6%/mM in groups 1–4, respectively. The difference in the regression coefficient was statistically significant between group 1 and 2 (F = 3.69, p < 0.05, ANCOVA), group 1 and 3 (F = 5.599, p < 0.05), and group 1 and 4 (F = 10.853, p < 0.005). This suggested that patients with higher basal iPTH responded poorly to modulate serum PTH levels by sensing the change of iCa. However, after an intensified oral calcitriol therapy to reduce iPTH, the PTH response in group 4 patients was restored to levels comparable with those observed in patients having lower basal iPTH. We also demonstrate that the ΔiPTH of patients of group 1 but not of other groups was significantly correlated with serum markers for bone metabolism; osteocalcin (r = 0.535, p < 0.05) and collagen type I C-terminal telopeptide (r = –0.575, p < 0.05). Conclusions: Our findings suggest that secretion of PTH is dynamically regulated by ΔiCa during hemodialysis and such oscillated PTH secretion may affect bone metabolism in a subset of dialysis patients.
Reciprocal Association of Plasma Adiponectin and Serum C-Reactive Protein Concentration in Haemodialysis Patients with End-Stage Kidney Disease – A Follow-Up Study
Authors: Ignacy Witold , Chudek Jerzy , Adamczak Marcin , Funahashi Tohru , Matsuzawa Yuji , Kokot Franciszek , Więcek Andrzej
Published: May 2005

Aim: Malnutrition and inflammation are involved in the pathogenesis of atherosclerosis and increased risk of cardiovascular diseases in end-stage renal disease. The aim of this study was to assess the relationship between concentrations of plasma adiponectin, serum C-reactive protein (CRP), carotid intima-media thickness (IMT) and duration of haemodialysis (HD) treatment in prevalent HD patients. Methods: Plasma adiponectin and serum CRP concentrations were estimated in 80 HD patients and 22 healthy controls. Carotid IMT was measured by ultrasound technique. HD patients were followed up for 23 ± 16 months. During this period, 24 of them died. Results: In HD patients, plasma adiponectin concentration was over 3 times higher than in controls (29.0 ± 2.1 vs. 8.7 ± 2.6 µg/ml; p < 0.001). HD patients with serum CRP concentrations ≧5 mg/l were characterized by a lower plasma adiponectin concentration than HD patients with the CRP <5 mg/l (23.9 ± 3.5 vs. 33.0 ± 3.1 µg/ml; p = 0.03). Plasma adiponectin and serum CRP concentrations were inversely related in HD (τ = –0.181; p = 0.02). No relationship between adiponectinaemia and IMT was observed. Survival (Kaplan-Meier analysis) within the lowest plasma adiponectin tertile tended (p = 0.06) to be the worse. Conclusions: (1) Inflammatory processes are associated with an inadequate low plasma adiponetin concentration in HD patients, and (2) a lower plasma adiponectin concentration seems to be a new predictor of poor outcome in HD patients.
Treatment with Cyclophosphamide in Elderly-Onset Nephrotic Syndrome
Authors: Mak Siu-ka , Lo Kin-yee , Wong Calvin Y. , Tong Gensy M.W. , Wong Ping-nam , Wong Andrew K.M.
Published: May 2005

Background:The best treatment of elderly-onset nephrotic syndrome has not been well defined. The use of corticosteroids or combination immunosuppressants may be associated with a significant incidence of side effects in the elderly. There is little data on the use of cyclophosphamide alone. Methods:We retrospectively reviewed 30 patients with idiopathic elderly-onset nephrotic syndrome treated with cyclophosphamide. Results:Male to female ratio was 2:1, mean age at diagnosis was 72.7 ± 5.9 years and average length of follow-up was 41.4 ± 21.3 months. Significant co-morbidities, including hypertension, were present in 57%. A raised serum creatinine level was found in 57%. Biopsy revealed 15 membranous nephropathy, 4 mesangial proliferative Gn, 5 IgA nephropathy, 3 minimal change nephropathy, 2 focal segmental glomerulosclerosis and 1 C1q nephropathy. Cyclophosphamide was given for 32.0 ± 16.2 weeks with an averaged cumulative dose per patient 177 ± 84 mg/kg BW. Remission (complete or partial) was attained by 40, 63, 80 and 87% of patients within 12, 24, 36 and 48 weeks of treatment, respectively. Eighteen patients attained complete remission and 9 partial remission after treatment. The mean interval to attain first remission (complete or partial) was 18.9 ± 14.6 weeks. This was not affected by age (p = NS) or initial albumin level (p = NS). At the time of last follow-up, all but 2 patients were in complete or partial remission with raised serum creatinine levels in 40%. Conclusions:Cyclophosphamide was effective and well tolerated in the treatment of elderly-onset nephrotic syndrome, with sustained remission and preserved renal function.
Risk Factors for Early Failure of Native Arteriovenous Fistulas
Authors: Ernandez T. , Saudan P. , Berney T. , Merminod T. , Bednarkiewicz M. , Martin P.-Y.
Published: May 2005

Introduction: Current guidelines recommend native arteriovenous fistulas (AVF) as the vascular access of choice for hemodialysis on account of the lower incidence of complications. However, this kind of vascular access has a high rate of early failure (early thrombosis or non-maturation). Our aim was to examine whether clear risk factors for early AVF failure could be identified in our patients. Subjects and Methods: Data of all patients who underwent creation of an AVF at the Geneva University Hospital from January 1998 to December 2002 were reviewed. Early failure was defined as a non-functioning fistula (thrombosis or absence of fistula maturation). Results: 119 patients underwent the creation of 148 native AVF, 88 (59.5%) in the forearm and 60 (40.5%) in the upper arm. 48 (32.4%) fistulae were created in diabetic patients. In a multiple logistic regression analysis, significant predictive factors of early failure were a distal location (adjusted odds ratio (aOR) = 8.21, 95% CI = 2.63–25.63, p < 0.001), female gender (aOR = 4.04, 95% CI = 1.44–11.30, p = 0.008), level of surgical expertise (aOR = 3.97, 95% CI = 1.39–11.32, p = 0.010) and diabetes mellitus (aOR = 3.19, 95% CI = 1.17–8.71, p = 0.024). Conclusion: Early failure of AVF occurs mainly in forearm sites among women and diabetic patients. Surgical expertise has also a significant influence. These results suggest that selection of a distal site for a native AVF has to be rigorously made for women and diabetic patients and that surgeon’s dedication is of primary importance to avoid early AVF failure occurrence.
Effects of Cisplatin, 5-Fluorouracil, and Radiation on Cell Cycle Regulation and Apoptosis in the Hypopharyngeal Carcinoma Cell Line
Authors: Lee Byung-Joo , Chon Kyong-Myong , Kim You-Sun , An Won G. , Roh Hwan-Jung , Goh Eui-Kyung , Wang Soo-Geun
Published: May 2005

In head and neck cancer including hypopharyngeal cancer, cisplatin and 5-fluorouracil (5-FU) usually have been used as neoadjuvant chemotherapeutic agents. We investigated the effects of cisplatin, 5-FU and radiation on p53 protein expression and cell responses (cell cycle arrest and/or apoptosis) in the hypopharyngeal carcinoma cell line (PNUH-12; mutant type p53). PNUH-12 cells were treated with cisplatin, 5-FU and radiation. The changes in the cells were assessed by a cell cytotoxicity assay, Western blotting (p53 and p21WAF1/CIP1 proteins), a DNA fragmentation assay, propidium iodide (PI) staining and DNA flow cytometry. The expression of p53 protein was increased after treatment with cisplatin and 5-FU, but not radiation. The expression of p21WAF1/CIP1 protein was increased only after treatment with 5-FU, not cisplatin or radiation. With cisplatin and radiation, we observed apoptosis both by DNA fragmentation and PI staining and increased S phase in cisplatin and G2 phase in radiation by DNA flow cytometry. But, with 5-FU, we could not observe apoptosis by DNA fragmentation and PI stain but only an increased G1 phase by DNA flow cytometry. In PNUH-12, radiation induced p53-independent apoptosis and p21WAF1/CIP1-independent G2-phase cell cycle arrest. Cisplatin induced p53-dependent apoptosis and p21WAF1/CIP1-independent S-phase cell cycle arrest and 5-FU induced p53 and p21WAF1/CIP1-dependent G1-phase cell cycle arrest, not apoptosis. Cisplatin and 5-FU induced p53-dependent pathways, but radiation p53-independent pathway. The cell responses by cisplatin, 5-FU and radiation were all different pathways.
Endothelial nitric oxide synthase plays a minor role in inhibition of arterial thrombus formation
Authors: Özüyaman Burcin , Küsters Susanne , Kirchhoff Elisabeth , Scharf Rüdiger , Schrader Jürgen , Gödecke Axel
Published: May 2005

Endothelial NO synthase (eNOS) expressed in the vascular en-dothelium or formed within platelets was postulated to inhibit platelet activation and aggregation. We have assessed the role of eNOS in platelet aggregation in vitro and in vivo by comparison of WT and eNOS-/- mice. Aggregometer studies revealed that collagen over a concentration range of 0.36–10 µg aggregated WT and eNOS-/- platelets to the same extent (10 µg: WT 86.7±4.7%, eNOS-/- 91±12%, n=6). Collagen treatment did not result in a significant increase in cGMP formation and VASP phosphorylation. Thrombin-induced P-selectin surface expression was unchanged in eNOS-/- platelets. In line with these findings no eNOS protein was detectable within the platelets of WT mice. In vivo, bleeding time after tail tip resection tended to be shorter in eNOS/- mice (WT: 116±35 s; eNOS-/- 109±37 s, n.s). Similarly, time to occlusion of the A.carotis after focal induction of thrombosis was 501±76 s (WT) and 457±95 s (eNOS-/-) (n.s.). These data demonstrate that eNOS-deficiency minimally affects platelet aggregation and is not associated with accelerated arterial thrombosis in vivo. Thus, in the mouse endothelial NO synthase does not play a major role in the autocrine modulation of platelet function and in thrombosis of conduit vessels in vivo.
Novel aberrant splicings caused by a splice site mutation (IVS1a+5g>a) in F7 gene
Authors: Fu Qihua , Wang Xuefeng , Hu Yiqun , Wang Zhenyi , Ding Qiulan , Wu Wenman , Wang Hongli
Published: May 2005

Low FVII coagulant activity (FVII:C 8.2%) and antigen level (FVII:Ag 34.1%) in a 46-year-old Chinese male led to a diagnosis of coagulation factor VII (FVII) deficiency. Compound heterozygous mutations were identified in his F7 gene:a G to A transition in the 5’ donor splice site of intron 1a (IVS1a+5g>a) and a T to G transition at the nucleotide position 10961 in exon 8, resulting in a His to Gln substitution at amino acid residue 348. An analysis of ectopic transcripts of F7 in the leukocytes of the patient reveals that the mutation (IVS1a+5g>a) is associated with two novel aberrant patterns of splicing. The predominant alternative transcript removes exon 2, but retains intron 3, which shifts the reading frame and predicts a premature translation termination at the nucleotide positions 2–4 in intron 3. The minor alternative transcript skips both exon 2 and exon 3 (FVII Δ2, 3), leading to an in-frame deletion of the propeptide and γ-carboxylated glutamic acid (Gla) domains of mature FVII protein. In vitro expression studies of the alternative transcript FVII Δ2, 3 by transient transfection of HEK 293 cells with PcDNA 3.1(-) expression vector showed that although the mutant protein could be secreted, no pro-coagulation activity was detected. The coexistence of the two abnormal transcripts and a heterozygous mutation His348Gln, explained the patient’s phenotype.
Baseline associations between postmenopausal hormone therapy and inflammatory, haemostatic, and lipid biomarkers of coronary heart disease
Authors: Pradhan Aruna , Lewis Cora , Manson JoAnn , Rossouw Jacques , Hendrix Susan , LaCroix Andrea , Ridker Paul , Langer Robert
Published: May 2005

Clinical trials of postmenopausal hormone therapy (PHT) have found an early increase in cardiovascular events, and have not demonstrated the reduction in coronary heart disease (CHD) predicted from changes in conventional risk factors or found in observational studies, suggesting that PHT may increase coronary risk through other pathways. We compared baseline levels of C-reactive protein (CRP), interleukin-6 (IL-6), sICAM-1, tissue plasminogen activator antigen (tPA-antigen), D-dimer, homocysteine, triglycerides, total-, HDL- and LDL- cholesterol in 304 cases with incident CHD and 304 controls, according to self-reported use of PHT. Subjects were selected from the 75,343 participants in the WHI Observational Study without baseline cardiovascular disease or cancer. PHT was associated with higher CRP, HDL and triglycerides, and lower tPA-antigen and homo-cysteine. CRP was highest in users of unopposed conjugated equine estrogen. Levels of IL-6, sICAM-1, D-dimer and total cholesterol did not differ between PHT users and non-users. Trans-dermal estrogen users had low levels of D-dimer and CRP. Among users of estrogen plus progestin (EP), CRP, IL-6, tPA-antigen, D-dimer, total cholesterol and triglycerides were higher in women with incident coronary events than controls. Estrogen alone (E) controls shared only the tPA-antigen association, but had higher HDL and lower LDL than E cases. In non-users CRP, tPA-antigen and D-dimer were associated with incident CHD. In summary, risk markers differed by PHT category. Some associations differed between women with and without incident CHD, especially for EP, where inflammatory and thrombotic markers were higher in cases. These associations remain speculative pending confirmation in randomized trials.