bluuebunny/crossref_metadata_2025 huggingface dataset:
Nitric oxide increases toxicity of hydrogen peroxide against rat liver endothelial cells and hepatocytes by inhibition of hydrogen peroxide degradation
Authors: Rauen Ursula , Li Tongju , Ioannidis Iosif , de Groot Herbert
Published: December 2006

Nitric oxide (NO) and hydrogen peroxide (H2O2) show cooperativity in their cytotoxic action. The present study was performed to decipher the mechanisms underlying this phenomenon. In cultured liver endothelial cells and in cultured, glutathione-depleted hepatocytes, the combined exposure to NO (released by spermine NONOate, 1 mM) and H2O2 (released by glucose oxidase) induced cell injury that was far higher than the injury elicited by NO or H2O2 alone. In both cell types, the addition of the NO donor increased H2O2 steady-state levels, although with different kinetics: in hepatocytes, the increase in H2O2 levels was already evident at early time points while in liver endothelial cells it became evident after ≥2 h of incubation. NO exposure inhibited H2O2 degradation, assessed after addition of 50 μM, 200 μM, or 4 mM authentic H2O2, significantly in both cell types. However, again, early and delayed inhibition was observed. The late inhibition of H2O2 degradation in endothelial cells was paralleled by a decrease in glutathione peroxidase activity. Glutathione peroxidase inactivation was prevented by hypoxia or by ascorbate, suggesting inactivation by reactive nitrogen oxide species (NOx). Early inhibition of H2O2 degradation by NO, in contrast, could be mimicked by the catalase inhibitor azide. Together, these results suggest that the cooperative effect of NO and H2O2 is due to inhibition of H2O2 degradation by NO, namely to inhibition of catalase by NO itself (predominant in hepatocytes) and/or to inhibition of glutathione peroxidase by NOx (prevailing in endothelial cells).
Upregulation of Na+/Ca2+ exchanger contributes to the enhanced Ca2+ entry in pulmonary artery smooth muscle cells from patients with idiopathic pulmonary arterial hypertension
Authors: Zhang Shen , Dong Hui , Rubin Lewis J. , Yuan Jason X.-J.
Published: December 2006

A rise in cytosolic Ca2+ concentration ([Ca2+]cyt) in pulmonary artery smooth muscle cells (PASMC) is a trigger for pulmonary vasoconstriction and a stimulus for PASMC proliferation and migration. Multiple mechanisms are involved in regulating [Ca2+]cyt in human PASMC. The resting [Ca2+]cyt and Ca2+ entry are both increased in PASMC from patients with idiopathic pulmonary arterial hypertension (IPAH), which is believed to be a critical mechanism for sustained pulmonary vasoconstriction and excessive pulmonary vascular remodeling in these patients. Here we report that protein expression of NCX1, an NCX family member of Na+/Ca2+ exchanger proteins is upregulated in PASMC from IPAH patients compared with PASMC from normal subjects and patients with other cardiopulmonary diseases. The Na+/Ca2+ exchanger operates in a forward (Ca2+ exit) and reverse (Ca2+ entry) mode. By activating the reverse mode of Na+/Ca2+ exchange, removal of extracellular Na+ caused a rapid increase in [Ca2+]cyt, which was significantly enhanced in IPAH PASMC compared with normal PASMC. Furthermore, passive depletion of intracellular Ca2+ stores using cyclopiazonic acid (10 μM) not only caused a rise in [Ca2+]cyt due to Ca2+ influx through store-operated Ca2+ channels but also mediated a rise in [Ca2+]cyt via the reverse mode of Na+/Ca2+ exchange. The upregulated NCX1 in IPAH PASMC led to an enhanced Ca2+ entry via the reverse mode of Na+/Ca2+ exchange, but did not accelerate Ca2+ extrusion via the forward mode of Na+/Ca2+ exchange. These observations indicate that the upregulated NCX1 and enhanced Ca2+ entry via the reverse mode of Na+/Ca2+ exchange are an additional mechanism responsible for the elevated [Ca2+]cyt in PASMC from IPAH patients.
Class A scavenger receptor-mediated cell adhesion requires the sequential activation of Lyn and PI3-kinase
Authors: Nikolic Dejan M. , Cholewa Jill , Gass Cecelia , Gong Ming C. , Post Steven R.
Published: December 2006

Class A scavenger receptors (SR-A) participate in multiple macrophage functions including macrophage adhesion to modified proteins. SR-A-mediated adhesion may therefore contribute to chronic inflammation by promoting macrophage accumulation at sites of protein modification. The mechanisms that couple SR-A binding to modified proteins with increased cell adhesion have not been defined. In this study, SR-A expressing HEK cells and SR-A+/+ or SR-A−/− macrophages were used to delineate the signaling pathways required for SR-A-mediated adhesion to modified protein. Inhibiting Gi/o activation, which decreases initial SR-A-mediated cell attachment, did not prevent the subsequent spreading of attached cells. In contrast, inhibition of Src kinases or PI3-kinase abolished SR-A-dependent cell spreading without affecting SR-A-mediated cell attachment. Consistent with these results, the Src kinase Lyn and PI3-kinase were sequentially activated during SR-A-mediated cell spreading. Furthermore, activation of both Lyn and PI3-kinase was required for enhancing paxillin phosphorylation. Activation of a Src kinase-PI3-kinase-Akt pathway was also observed in cells expressing a truncated SR-A protein that does not internalize indicating that SR-A-mediated activation of intracellular signaling cascades following adhesion to MDA-BSA is independent of receptor internalization. Thus SR-A binding to modified protein activates signaling cascades that have distinct roles in regulating initial cell attachment and subsequent cell spreading.
Identification of a novel interaction between the Ca2+-binding protein S100A11 and the Ca2+- and phospholipid-binding protein annexin A6
Authors: Chang Ning , Sutherland Cindy , Hesse Eva , Winkfein Robert , Wiehler William B. , Pho Mark , Veillette Claude , Li Susan , Wilson David P. , Kiss Enikõ , Walsh Michael P.
Published: December 2006

S100A11 is a member of the S100 family of EF-hand Ca2+-binding proteins, which is expressed in smooth muscle and other tissues. Ca2+ binding to S100A11 induces a conformational change that exposes a hydrophobic surface for interaction with target proteins. Affinity chromatography with immobilized S100A11 was used to isolate a 70-kDa protein from smooth muscle that bound to S100A11 in a Ca2+-dependent manner and was identified by mass spectrometry as annexin A6. Direct Ca2+-dependent interaction between S100A11 and annexin A6 was confirmed by affinity chromatography of the purified bacterially expressed proteins, by gel overlay of annexin A6 with purified S100A11, by chemical cross-linking, and by coprecipitation of S100A11 with annexin A6 bound to liposomes. The expression of S100A11 and annexin A6 in the same cell type was verified by RT-PCR and immunocytochemistry of isolated vascular smooth muscle cells. The site of binding of S100A11 on annexin A6 was investigated by partial tryptic digestion and deletion mutagenesis. The unique NH2 terminal head region of annexin A6 was not required for S100A11 binding, but binding sites were identified in both NH2- and COOH-terminal halves of the molecule. We hypothesize that an agonist-induced increase in cytosolic free [Ca2+] leads to formation of a complex of S100A11 and annexin A6, which forms a physical connection between the plasma membrane and the cytoskeleton, or plays a role in the formation of signaling complexes at the level of the sarcolemma.
Intracellular cAMP: the “switch” that triggers on “spontaneous transient outward currents” generation in freshly isolated myocytes from thoracic aorta
Authors: Hayoz Sébastien , Bény Jean-Louis , Bychkov Rostislav
Published: December 2006

Spontaneous transient outward currents (STOCs) have been reported in resistance and small arteries but have not yet been found in thoracic aorta. Do thoracic aorta myocytes possess cellular machinery that generates STOCs? It was found that the majority of aortic myocytes do not generate STOCs. STOCs were generated in 8.7% of freshly isolated aortic myocytes. Myocytes that did not generate STOCs we have called “silent” myocytes and myocytes with STOCs have been called “active.” STOCs recorded in active myocytes were voltage dependent and were inhibited by ryanodine, caffeine, and charybdotoxin. Forskolin was reported to increase STOCs frequency in myocytes isolated from resistance arteries. Forskolin (10 μM) triggered STOCs generation in 35.1% of silent aortic myocytes. In 36.8% percent of silent myocytes, forskolin did not trigger STOCs but increased the amplitude of charybdotoxin-sensitive outward net current to 136.1 ± 8.5% at 0 mV. Membrane-permeable 8BrcAMP triggered STOCs generation in 38.7% of silent myocytes. Forskolin- or 8BrcAMP-triggered STOCs were inhibited by charybdotoxin. 8BrcAMP also increased open probability of BKCachannels in BAPTA-AM-pretreated cells. Our data demonstrate that, in contrast to resistance arteries, STOCs are present just in the minority of myocytes in the thoracic aorta. However, cellular machinery that generates STOCs can be “switched” on by cAMP. Such an inactive cellular mechanism could modulate the contractility of the thoracic aorta in response to physiological demand.
Hypertonicity triggers RhoA-dependent assembly of myosin-containing striated polygonal actin networks in endothelial cells
Authors: Malek Adel M. , Xu Chang , Kim Edward S. , Alper Seth L.
Published: December 2006

Endothelial cells respond to mechanical stresses of the circulation with cytoskeletal rearrangements such as F-actin stress fiber alignment along the axis of fluid flow. Endothelial cells are exposed to hypertonic stress in the renal medulla or during mannitol treatment of cerebral edema. We report here that arterial endothelial cells exposed to hypertonic stress rearranged F-actin into novel actin-myosin II fibers with regular 0.5-μm striations, in which α-actinin colocalizes with actin. These striated fibers assembled over hours into three-dimensional, irregular, polygonal actin networks most prominent at the cell base, and occasionally surrounding the nucleus in a geodesic-like structure. Hypertonicity-induced assembly of striated polygonal actin networks was inhibited by cytochalasin D, blebbistatin, cell ATP depletion, and intracellular Ca2+chelation but did not require intact microtubules, regulatory volume increase, or de novo RNA or protein synthesis. Striated polygonal actin network assembly was insensitive to inhibitors of MAP kinases, tyrosine kinases, or phosphatidylinositol 3-kinase, but was prevented by C3 exotoxin, by the RhoA kinase inhibitor Y-27632, and by overexpressed dominant-negative RhoA. In contrast, overexpression of dominant-negative Rac or of dominant-negative cdc42 cDNAs did not prevent striated polygonal actin network assembly. The actin networks described here are novel in structure, as striated actin-myosin structures in nonmuscle cells, as a cellular response to hypertonicity, and as a cytoskeletal regulatory function of RhoA. Endothelial cells may use RhoA-dependent striated polygonal actin networks, possibly in concert with cytoskeletal load-bearing elements, as a contractile, tension-generating component of their defense against isotropic compressive forces.
Thyroid hormone inhibits biliary growth in bile duct-ligated rats by PLC/IP3/Ca2+-dependent downregulation of SRC/ERK1/2
Authors: Fava Giammarco , Ueno Yoshiyuki , Glaser Shannon , Francis Heather , DeMorrow Sharon , Marucci Luca , Marzioni Marco , Benedetti Antonio , Venter Julie , Vaculin Bradley , Vaculin Shelley , Alpini Gianfranco
Published: December 2006

The role of the thyroid hormone agonist 3,3′,5 l-tri-iodothyronine (T3) on cholangiocytes is unknown. We evaluated the in vivo and in vitro effects of T3 on cholangiocyte proliferation of bile duct-ligated (BDL) rats. We assessed the expression of α1-, α2-, β1-, and β2-thyroid hormone receptors (THRs) by immunohistochemistry in liver sections from normal and BDL rats. BDL rats were treated with T3 (38.4 μg/day) or vehicle for 1 wk. We evaluated 1) biliary mass and apoptosis in liver sections and 2) proliferation in cholangiocytes. Serum-free T3 levels were measured by chemiluminescence. Purified BDL cholangiocytes were treated with 0.2% BSA or T3 (1 μM) in the absence/presence of U-73122 (PLC inhibitor) or BAPTA/AM (intracellular Ca2+ chelator) before measurement of PCNA protein expression by immunoblots. The in vitro effects of T3 (1 μM) on 1) cAMP, IP3, and Ca2+ levels and 2) the phosphorylation of Src Tyr139 and Tyr530 (that, together, regulate Src activity) and ERK1/2 of BDL cholangiocytes were also evaluated. α1-, α2-, β1-, and β2-THRs were expressed by bile ducts of normal and BDL rats. In vivo, T3 decreased cholangiocyte proliferation of BDL rats. In vitro, T3 inhibition of PCNA protein expression was blocked by U-73122 and BAPTA/AM. Furthermore, T3 1) increased IP3 and Ca2+ levels and 2) decreased Src and ERK1/2 phosphorylation of BDL cholangiocytes. T3 inhibits cholangiocyte proliferation of BDL rats by PLC/IP3/Ca2+-dependent decreased phosphorylation of Src/ERK1/2. Activation of the intracellular signals triggered by T3 may modulate the excess of cholangiocyte proliferation in liver diseases.
Long-term feeding of various fat diets modulates azoxymethane-induced colon carcinogenesis through Wnt/β-catenin signaling in rats
Authors: Fujise Takehiro , Iwakiri Ryuichi , Kakimoto Takashi , Shiraishi Ryosuke , Sakata Yasuhisa , Wu Bin , Tsunada Seiji , Ootani Akifumi , Fujimoto Kazuma
Published: December 2006

The Wnt signaling pathway plays an essential role in carcinogenesis, and the amount of fat intake and composition of dietary fatty acids are crucial factors for colon carcinogenesis. We investigated whether various dietary fats affected the Wnt signaling pathway of colon tumorigenesis in azoxymethane (AOM)-treated rats. Male Sprague-Dawley rats were given intraperitoneal injections of AOM and supplemented with 10% corn, olive, beef, and fish oil for 44 wk. Aberrant crypt foci (ACF) and tumors were examined at 12 and 44 wk. Normal appearing colon mucosal proliferation and apoptosis were evaluated by 5-bromo-2′-deoxyuridine (BrdU) incorporation and percentages of fragmented DNA, respectively. Expressions of β-catenin, cyclin D1, Wnt2, Wnt3, and Wnt5a of normal appearing colon mucosa were analyzed by Western blot analysis. Long-term dietary corn oil and beef tallow increased ACF, tumor incidence, and tumor numbers in AOM-treated rats. In contrast, both olive and fish oil inhibited them. Dietary corn oil and beef tallow increased BrdU incorporation and the expression of cytosolic β-catenin and cyclin D1 and decreased apoptosis in the colon mucosa. Expressions of Wnt2 and Wnt3 in rats fed with beef tallow and Wnt5a in rats fed with corn oil increased with or without AOM-treatment. BrdU-incorporated cells were often observed at the tops of crypts in rats fed with beef tallow, whereas this was not observed in rats fed with the other diet. Long-term high intake of corn oil and beef tallow enhanced cell proliferation through Wnt signaling and modulated the distribution of proliferating cells, which might contribute to promoting effects in colon tumorigenesis.
Regulation of Triglyceride Metabolism. III. Emerging role of lipid droplet protein ADFP in health and disease
Authors: Chang Benny Hung-Junn , Chan Lawrence
Published: December 2006

ADFP is the major lipid droplet protein present in all cells that accumulate lipids either normally or abnormally. Although discovered about 15 years ago, it was only in the last few years that we began to have a working knowledge of its possible role in lipid droplet homeostasis at the whole organism level. In this perspective, we concentrate on the potential function of ADFP in various tissues. Space limitation has precluded a complete cataloging of all publications on the topic. We instead highlight some of the salient developments in the last few years.
Enhanced pelvic responses to stressors in female CRF-overexpressing mice
Authors: Million M. , Wang L. , Stenzel-Poore M. P. , Coste S. C. , Yuan P. Q. , Lamy C. , Rivier J. , Buffington T. , Taché Y.
Published: December 2006

Acute stress affects gut functions through the activation of corticotropin-releasing factor (CRF) receptors. The impact of acute stress on pelvic viscera in the context of chronic stress is not well characterized. We investigated the colonic, urinary, and locomotor responses monitored as fecal pellet output (FPO), urine voiding, and ambulatory activity, respectively, in female and male CRF-overexpressing (CRF-OE) mice, a chronic stress model, and their wild-type littermates (WTL). Female CRF-OE mice, compared with WTL, had enhanced FPO to 2-min handling (150%) and 60-min novel environment (155%) but displayed a similar response to a 60-min partial restraint stress. Female CRF-OE mice, compared with WTL, also had a significantly increased number of urine spots (7.3 ± 1.4 vs. 1.3 ± 0.8 spots/h) and lower locomotor activity (246.8 ± 47.8 vs. 388.2 ± 31.9 entries/h) to a novel environment. Male CRF-OE mice and WTL both responded to a novel environment but failed to show differences between them in colonic and locomotor responses. Male WTL, compared with female WTL, had higher FPO (113%). In female CRF-OE mice, the CRF1/CRF2 receptor antagonist astressin B and the selective CRF2 receptor agonist mouse urocortin 2 (injected peripherally) prevented the enhanced defecation without affecting urine or locomotor responses to novel environment. RT-PCR showed that CRF1 and CRF2 receptors are expressed in the mouse colonic tissues. The data show that chronic stress, due to continuous central CRF overdrive, renders female CRF-OE mice to have enhanced pelvic and altered behavioral responses to superimposed mild stressors and that CRF1-initiated colonic response is counteracted by selective activation of CRF2 receptor.
Physiologically high concentrations of 17β-estradiol enhance NF-κB activity in human T cells
Authors: Hirano Suzue , Furutama Daisuke , Hanafusa Toshiaki
Published: December 2006

Estrogen has diverse effects on inflammation and immune responses. That pregnancy is associated with remission of some autoimmune diseases and exacerbation of others suggests that physiological fluctuation in estrogen levels could affect the immune responses in humans. However, the molecular basis for these phenomena is poorly understood. We hypothesized that fluctuations of estrogen levels modulate intracellular signaling for immune responses via estrogen receptors (ERs). In reporter assays, 17β-estradiol (E2) at a physiologically high concentration increased the activity of NF-κB in Jurkat cells stimulated by PMA/ionomycin or TNF-α. Overexpression and RNA interference experiments suggested that the effects were mediated through ERβ. Immunoprecipitation assay showed that both ERα and ERβ are directly associated with NF-κB in the cell nucleus. Using chromatin immunoprecipitation assay, we confirmed that ERα and ERβ associated with NF-κB and steroid hormone coactivators at the promoter region of NF-κB regulated gene. Considering that NF-κB regulates the expression of various genes essential for cell growth and death, estrogen could regulate the fate of T cells by affecting the activity of NF-κB. To determine whether E2 alters the fate of T cells, we investigated E2 actions on T cell apoptosis, a well-known NF-κB-mediated phenomenon. E2 increased apoptosis of Jurkat cells and decreased that of human peripheral blood T cells. Our results indicate that E2 at a physiologically high concentration modulates NF-κB signaling in human T cells via ERβ and affects T cell survival, suggesting that these actions may underlie the gender differences in autoimmune diseases.
The melting of pulmonary surfactant monolayers
Authors: Yan Wenfei , Biswas Samares C. , Laderas Ted G. , Hall Stephen B.
Published: December 2006

Monomolecular films of phospholipids in the liquid-expanded (LE) phase after supercompression to high surface pressures (π), well above the equilibrium surface pressure (πe) at which fluid films collapse from the interface to form a three-dimensional bulk phase, and in the tilted-condensed (TC) phase both replicate the resistance to collapse that is characteristic of alveolar films in the lungs. To provide the basis for determining which film is present in the alveolus, we measured the melting characteristics of monolayers containing TC dipalmitoyl phosphatidylcholine (DPPC), as well as supercompressed 1-palmitoyl-2-oleoyl phosphatidylcholine and calf lung surfactant extract (CLSE). Films generated by appropriate manipulations on a captive bubble were heated from ≤27°C to ≥60°C at different constant π above πe. DPPC showed the abrupt expansion expected for the TC-LE phase transition, followed by the contraction produced by collapse. Supercompressed CLSE showed no evidence of the TC-LE expansion, arguing that supercompression did not simply convert the mixed lipid film to TC DPPC. For both DPPC and CLSE, the melting point, taken as the temperature at which collapse began, increased at higher π, in contrast to 1-palmitoyl-2-oleoyl phosphatidylcholine, for which higher π produced collapse at lower temperatures. For π between 50 and 65 mN/m, DPPC melted at 48–55°C, well above the main transition for bilayers at 41°C. At each π, CLSE melted at temperatures >10°C lower. The distinct melting points for TC DPPC and supercompressed CLSE provide the basis by which the nature of the alveolar film might be determined from the temperature-dependence of pulmonary mechanics.
It May Take Inflammation, Phosphorylation and Ubiquitination to ‘Tangle’ in Alzheimer’s Disease
Authors: Arnaud Lisette , Robakis Nikolaos K. , Figueiredo-Pereira Maria E.
Published: December 2006

Neurofibrillary tangles (NFT) are one of the pathologic hallmarks of Alzheimer’s disease (AD). Their major component is tau, a protein that becomes hyperphosphorylated and accumulates into insoluble paired helical filaments. During the course of the disease such filaments aggregate into bulky NFT that get ubiquitinated. What triggers their formation is not known, but neuroinflammation could play a role. Neuroinflammation is an active process detectable in the earliest stages of AD. The neuronal toxicity associated with inflammation makes it a potential risk factor in the pathogenesis of chronic neurodegenerative diseases, such as AD. Determining the sequence of events that lead to this devastating disease has become one of the most important goals for AD prevention and treatment. In this review we focus on three topics relevant to AD pathology and to NFT formation: (1) what triggers CNS inflammation resulting in glia activation and neuronal toxicity; (2) how products of inflammation might change the substrate specificity of kinases/phosphatases leading to tau phosphorylation at pathological sites; (3) the relationship between the ubiquitin/proteasome pathway and tau ubiquitination and accumulation in NFT. The overall aim of this review is to provide a challenging and sometimes provocative survey of important contributions supporting the view that CNS inflammation might be a critical contributor to AD pathology. Neuronal cell death resulting from neuroinflammatory processes may have devastating effects as, in the vast majority of cases, neurons lost to disease cannot be replaced. In order to design therapies that will prevent endangered neurons from dying, it is critical that we learn more about the effects of neuroinflammation and its products.
Successful Microsurgical Penile Replantation after a Workplace Injury
Authors: Fuller Andrew , Bolt John , Carney Bernard
Published: December 2006

Microsurgical techniques offer the opportunity for successful replantation of the amputated penis. We report the case of a patient who sustained a traumatic penile amputation as a result of a workplace accident. Microsurgical replantation was achieved with combined urological and plastic surgical input.
Role of Nitric Oxide in Urinary Bladder Function: Effect of L-Arginine
Authors: Whitbeck Catherine , Chichester Paul , Sokol Rebekah , Levin Robert M.
Published: December 2006

Background: Evidence indicates that decreased blood flow to the bladder plays a major role in obstructive bladder dysfunction in the rabbit model of partial bladder outlet obstruction (PBOO), and that nitric oxide (NO) regulation of blood flow may be important in modulating the degree of obstructive bladder dysfunction. The specific aim of our study is to determine the effect of feeding rabbits a diet high in L-arginine on the response to PBOO. Materials and Methods: Sixteen male NZ White rabbits were separated into 4 groups of 4 each. The rabbits in groups 1 and 3 underwent PBOO. The rabbits in groups 2 and 4 were sham-operated. For 1 week prior to surgery, and 2 weeks postoperatively, each rabbit in groups 1 and 2 was put on a diet containing 7% arginine. Rabbits in groups 3 and 4 were on a normal diet (0.76% arginine). Results: PBOO resulted in a greater increase in bladder weight in the control group than the arginine group. PBOO resulted in a greater decrease in compliance in the control group than the arginine group. The contractile responses to all agents in the arginine control group were greater than in the control normal diet group. PBOO resulted in a greater decrease in the response to field stimulation in the control group than in the arginine group. Conclusions: These studies clearly demonstrate that feeding rabbits a diet high in L-arginine was beneficial for the control rabbits, and reduced the level of dysfunctions following PBOO.
Impact of Self-Perceived Bothersomeness on Health-Related Quality of Life in Women with Overactive Bladder Symptoms
Authors: Oh Seung-June , Ku Ja Hyeon
Published: December 2006

Introduction: The objective of this study was to examine the impact of self-perceived bothersomeness of overactive bladder (OAB) symptoms on the health-related quality of life (QOL). Patients andMethods: A total of 92 women with a mean age of 53.3 (range 23–79) years suffering from OAB were included in the study. The Medical Outcomes Study 36-Item Short-Form Health Survey (SF-36) and King’s Health Questionnaire (KHQ) were used to assess the QOL. The patients were divided into two groups according to the bothersomeness degree of OAB: ‘low’ and ‘moderate’ to ‘high’. Results: The SF-36 and the KHQ scores of the patients were significantly different from those of the controls (p <0.05), except for three domains of the SF-36. No difference in the results of the frequency-volume charts was observed between the two patient groups. The scores of the SF-36 and the KHQ domains did not correlate with the data of the frequency-volume charts. Significant differences were found between the two patient groups for most domains of the SF-36 (p <0.05). Significant differences were also detected in most domains of the KHQ (p <0.05). Conclusions: Objective data are not a sensitive tool for measuring the QOL in women with OAB symptoms. Our findings suggest that patient-perceived bothersomeness significantly influences QOL and that strategies for assessing bothersomeness should be developed to evaluate the QOL in these patients.
Aggressive Fibromatosis in the Urological System
Authors: Shi Benkang , Zhu Yaofeng , Xu Zhishun , Liu Yuqiang , Zheng Baozhong , Qi Tianwei
Published: December 2006

Aggressive fibromatoses (AF) are locally aggressive neoplasms that do not metastasize but are frequently associated with one or more recurrences and subsequent associated morbidity. AF in the urological system is quite rare and has mainly been described in single case reports or as isolated cases in a large series of extra-abdominal desmoid tumors. We report 2 cases of AF in the bladder and scrotum, and provide a review of similar published cases.
Estrogen Differentially Modulates the Cannabinoid- Induced Presynaptic Inhibition of Amino Acid Neurotransmission in Proopiomelanocortin Neurons of the Arcuate Nucleus
Authors: Nguyen Que Huong , Wagner Edward J.
Published: December 2006

The present study sought to determine whether cannabinoids inhibit glutamatergic and GABAergic synaptic input onto neurons of the hypothalamic arcuate nucleus (ARC), and whether estrogen modulates this process. Whole-cell patch clamp recordings were performed in hypothalamic slices prepared from ovariectomized female guinea pigs. CB1 receptor activation reduced the amplitude of excitatory postsynaptic currents (EPSCs) evoked by electrical stimulation that were sensitive to ionotropic glutamate receptor antagonists. The CB1 receptor antagonist AM251 increased evoked EPSC (eEPSC) amplitude, and reversed the agonist-induced decrease. CB1 receptor activation similarly decreased the amplitude of evoked inhibitory postsynaptic currents (eIPSCs). The cannabinoid-induced reduction in eEPSC and eIPSC amplitude correlated with a decrease in the frequency of miniature EPSCs (mEPSCs) and IPSCs (mIPSCs) that were abolished by ionotropic glutamate and GABAA receptor antagonists, respectively. AM251 increased mEPSC frequency, and antagonized the agonist-induced decrease. Compared to neurons obtained from vehicle-treated controls, estradiol benzoate (25 µg; s.c.) given 24 h prior to experimentation increased mEPSC frequency, and markedly decreased the potency of CB1 receptor agonists to decrease mEPSC frequency. Conversely, the steroid potentiated the cannabinoid-induced decrease in mIPSC frequency. These effects were observed in neurons subsequently identified as proopiomelanocortin (POMC) neurons. These data reveal that ARC neurons, including POMC neurons, receive glutamatergic and GABAergic synaptic inputs that are presynaptically inhibited by cannabinoids, and differentially modulated by estrogen. These opposing effects of estrogen on the cannabinoid regulation of amino acid neurotransmission excite POMC neurons, and lend additional insight into the mechanisms underlying estrogen-induced anorexia and negative feedback of the reproductive axis.
Evolution of the Diffuse Neuroendocrine System – Clear Cells and Cloudy Origins
Authors: Modlin Irvin M. , Champaneria Manish C. , Bornschein Jan , Kidd Mark
Published: December 2006

As early as the 2nd century, Galen proposed that ‘vital spirits’ in the blood regulated human bodily functions. However, the concept of hormonal activity required a further 18 centuries to develop and relied upon the identification of ‘ductless glands’, Schwann’s cell and the recognition by Bayliss and Starling of chemical messengers. Bernard’s introduction of ‘internal secretion’ and its role in homeostasis laid a physiological basis for the development of endocrinology. Kocher and Addison recognized the consequences of ablation of glands by disease or surgery and identified their necessary role in life. Detailed descriptions of the endocrine cells of the gut and pancreas and their putative function were provided by Heidenhain, Langerhans, Laguesse and Sharpey-Schäfer. Despite the dominant 19th century concept of nervism (Pavlov), in 1902, Starling and Bayliss using Hardy’s term ‘hormonos’ described secretin and in so doing, established the gut as an endocrine organ. Thus, nervism was supplanted by hormonal regulation of function and thereafter numerous bioactive gut peptides and amines were identified. At virtually the same time (1892), Ramón y Cajal of Madrid reported the existence of a group of specialized intestinal cells that he referred to as ‘interstitial cells’. Cajal postulated that they might function as an interface between the neural system and the smooth muscles of the gut. Some 22 years later, Keith suggested that their function might be analogous to the electroconductive system of the heart and proposed their role as components of an intestinal pacemaker system. This prescient hypothesis was subsequently confirmed in 1982 by Thuneberg and a decade later Maede identified c-Kit as a critical molecular regulator in the development and function of the interstitial cells of Cajal and further confirmed the commonality of neural and endocrine cells. The additional characterization of the endocrine regulatory system of the GI tract was implemented when Feyrter (1938) using Masson’s staining techniques, identified ‘helle Zellen’ within the pancreatic ductal system and the intestinal epithelium and proposed the concept of a diffuse neuroendocrine system. Pearse subsequently grouped the various cells belonging to that system under the rubric of a unifying APUD series. Currently, the gut neuroendocrine system is viewed as a syncytium of neural and endocrine cells sharing a common cell lineage whose phenotypic regulation is as yet unclear. Their key role in the regulation of gastrointestinal function is, however, indubitable.
Testosterone Inhibition of Growth Hormone Release Stimulated by a Growth Hormone Secretagogue
Authors: Rigamonti Antonello E. , Cella Silvano G. , Giordani Claudio , Bonomo Sara M. , Giunta Marialuisa , Sartorio Alessandro , Müller Eugenio
Published: December 2006

Anabolic steroids are frequently taken by athletes and bodybuilders together with recombinant human GH (rhGH), though there is some scientific evidence that the use of anabolic steroids reverses the rhGH-induced effects. Recently, we have shown that treatment with rhGH (0.2 IU/kg s.c., daily × 12 days) in the dog markedly reduced the canine GH (cGH) responses stimulated by EP51216, a GH secretagogue (GHS), evaluated after 3 and 5 daily rhGH injections, and that the inhibition was still present a few days after rhGH discontinuation. The aim of the present study was to evaluate in the dog the GH response to EP51216 (125 µg/kg i.v.) in a condition of enhanced androgenic function (i.e. acute injection or 15-day treatment with testosterone at the dose of 2 mg/kg i.m. on alternate days), and in the hypophysectomized rat the hypothalamic and hippocampal expression of ghrelin, the receptor of GHSs (GHS-R), GH-releasing hormone (GHRH) and somatostatin (SS) after specific hormonal replacement therapies (testosterone, 1 mg/kg/day s.c.; hydrocortisone, 500 µg/kg/day s.c.; rhGH, 400 µg/kg/day s.c.; 0.9% saline 0.1 ml/kg/day s.c.; ×11 days). In the dog experiments, under baseline conditions, a single injection of EP51216 elicited an abrupt rise of plasma cGH. Twenty-four hours from the acute bolus injection of testosterone, Cmax and AUC0–90 of the GHS-stimulated cGH response were significantly lower than baseline cGH response; 5 days later, there was still a significant decrease of either parameter versus the original values. Short-term treatment with testosterone markedly reduced the GHS-stimulated cGH responses evaluated during (5th bolus) and at the end (8th bolus) of testosterone treatment. Four and 8 days after testosterone withdrawal, the EP51216-stimulated cGH response was still significantly reduced when compared with that under baseline conditions. Plasma concentrations of insulin-like growth factor 1 (IGF-1) were stable until the 5th bolus of testosterone and decreased progressively in the remaining time of the testosterone treatment; 4 and 8 days from treatment withdrawal, IGF-1 levels were still suppressed. In rat studies, hypothalamic mRNA levels of GHS-R were significantly reduced by treatments with testosterone and hydrocortisone, whereas hippocampal expressions of ghrelin, GHRH and SS were reduced by rhGH replacement therapy. In conclusion, these studies show that a single administration of testosterone can abrogate the cGH response ensuing acute stimulation by a GHS; the inhibitory effect of testosterone on the cGH response to GHS is present during and even 8 days after termination of a short-lived treatment with testosterone; these events occur via a downregulation of hypothalamic GHS-R.
Insulin-Degrading Enzyme Haplotypes Affect Insulin Levels but Not Dementia Risk
Authors: Marlowe Lauren , Peila Rita , Benke Kelly Suzanne , Hardy John , White Lon R. , Launer Lenore J. , Myers Amanda
Published: December 2006

Background: Insulin-degrading enzyme (IDE) polymorphism is hypothesized to regulate insulin levels as well as processes involved in neuronal compromise found in dementia. Methods: We examined the association of IDE haplotypes with dementia and insulin levels in a single well-characterized cohort of Japanese-American men born between 1900 and 1919 and followed since 1965. In 1991, a fasting insulin was obtained; dementia cases were ascertained in 1991 and 1994 in a multi-stage procedure, diagnoses were made according to international guidelines. Five single-nucleotide polymorphisms were genotyped and used for haplotype analysis in a sample of 179 Alzheimer’s disease cases, 104 vascular dementia cases and 516 controls nested in the total cohort. Results: The global test for the haplotype effect on insulin levels was significant (p < 0.0001), adjusting for age, education, apolipoprotein Ε4 status and fasting glucose. Conclusion: There was no association of IDE haplotypes with the risk of dementia. This study suggests IDE may be indirectly related to dementia via its regulation of insulin levels, but it is not a major gene for Alzheimer’s.
Association of Tau Haplotype-Tagging Polymorphisms with Parkinson’s Disease in Diverse Ethnic Parkinson’s Disease Cohorts
Authors: Fung H.C. , Xiromerisiou G. , Gibbs J.R. , Wu Y.R. , Eerola J. , Gourbali V. , Hellström O. , Chen C.M. , Duckworth J. , Papadimitriou A. , Tienari P.J. , Hadjigeorgiou G.M. , Hardy J. , Singleton A.B.
Published: December 2006

Background: The overlap in the clinical and pathological features of tauopathies and synucleinopathies raises the possibility that the tau protein may be important in Parkinson’s disease (PD) pathogenesis. Several MAPT polymorphisms that define the tau H1 haplotype have been investigated for an association with PD with conflicting results; however, two meta-analyses support an association between haplotype H1 and PD. Methods: In this study, we recruited 508 patients and 611 healthy controls from Greek, Finnish and Taiwanese populations. We examined the possible genetic role of variation within MAPT in PD using haplotype-tagging single polymorphisms (SNPs) in these ethnically different PD populations. Results: We identified a moderate association at SNP rs3785883 in the Greek cohort for both allele and genotype frequency (p = 0.01, p = 0.05, respectively) as well as for SNP rs7521 (genotype p = 0.02) and rs242557 (p = 0.01 genotypic, p = 0.04 allelic) in the Finnish population. There were no significant differences in genotype or allele distribution between cases and controls in the Taiwanese cohort. Conclusion: We failed to demonstrate a consistent association between the MAPT H1 haplotype (delineated by intron 9 ins/del) and PD in three ethnically diverse populations. However, the data presented here suggest that subhaplotypes of haplotype H1 may confer susceptibility to PD, and that either allelic heterogeneity or different haplotype composition explain the divergent haplotype results.
Genetic Association Study on Colony-Stimulating Factor 1 in Alzheimer’s Disease
Authors: Wollmer M. Axel , Nitsch Roger M. , Hock Christoph , Papassotiropoulos Andreas
Published: December 2006

Background: Colony-stimulating factor 1 (CSF1) regulates the proliferation and differentiation of myelomonocytic cells. Microglial cells of CSF1-deficient mice are reduced in number and are functionally impaired. CSF1-deficient mice exhibit subtle neurodevelopmental defects, enhanced neuronal vulnerability. Moreover, it has been reported that these mice may have amyloid-plaque-like depositions in the brain at an early age. The human CSF1 gene maps to chromosome 1p21–p13, a region previously linked to Alzheimer’s disease (AD). Thus, CSF1 is a functional and positional candidate gene for AD. Objective: We assessed if genetic variability of CSF1 may influence the risk for AD. Methods: We conducted a population-based case-control association study with 3 single nucleotide polymorphisms (SNPs) across the CSF1 locus in a sample of n = 185 (rs3093054, rs756325) and n = 327 (rs1058885) individuals. Results: None of the 3 investigated SNPs was associated with the risk for AD in our sample. Conclusion: These data do not support the hypothesis that genetic variability of CSF1 influences the risk for AD.
Evidence for the Presence of Molecules Related to the Neuropeptide CGRP in Two Cephalopods, Sepia officinalis and Nautilus macromphalus: Comparison with Its Target Organ Distribution
Authors: Lafont Anne-Gaëlle , Dufour Sylvie , Fouchereau-Peron Martine
Published: December 2006

Calcitonin gene-related peptide (CGRP) is a neuropeptide mainly involved in brain and cardiovascular functions in mammals. We investigated its presence and potential roles in two cephalopods, Sepia officinalis and Nautilus macromphalus. CGRP-like, but not calcitonin (CT)-like, molecules were detected by specific radioimmuno- and radioreceptor assays in the brain, optic lobes, branchial heart or afferent branchial vein and kidney. Gel exclusion chromatography of cephalopod brain extracts, followed by SDS-PAGE, indicated that CGRP-like molecules had a molecular weight of around 3 kDa, close to that of human CGRP. The distribution of CGRP target organs was characterized by binding studies in cuttlefish. Specific CGRP binding sites were detected in the brain, optic lobes, and kidney, indicating potential autocrine/paracrine roles of CGRP. Specific CGRP binding sites were also detected in the gills and shell sac that do not contain the peptide itself, indicating potential endocrine roles of CGRP. Accordingly, high circulating levels of CGRP-like molecules were detected in hemolymph of both cuttlefish and nautilus, unlike the situation in mammals. CGRP binding sites were further characterized in the cuttlefish gills by the Scatchard method. Our study indicates that the brain neurotransmitter role of CGRP could represent an ancient role in metazoa, already present in cephalopods and conserved among vertebrates. In contrast, the endocrine role of CGRP, which was suggested in cephalopods and also present in teleosts, may have been lost during the evolution of the tetrapod lineage. Our data support the hypothesis that CGRP represents the ancestral molecule of the CT/CGRP family appeared in metazoa before the vertebrate emergence.
Effect of Fasting on Cocaine-Amphetamine-Regulated Transcript, Neuropeptide Y, and Leptin Receptor Expression in the Non-Human Primate Hypothalamus
Authors: Van Vugt Dean A. , Lujan Marla E. , Froats Mark , Krzemien Alicja , Couceyro Pastor R. , Reid Robert L.
Published: December 2006

Leptin is a cytokine produced by white adipose tissue that circulates in direct proportion to adiposity and is an important signal of energy balance. Leptin inhibits food intake in rodents by inhibiting the orexigenic neuropetides neuropeptide Y (NPY) and agouti regulated peptide (AgRP) and stimulating the anorexigenic neuropeptides α-melanocyte-stimulating hormone (α-MSH) and cocaine-amphetamine-regulated transcript (CART). In order to extend our understanding of neuroendocrine regulation of appetite in the primate, we determined the effect of a metabolic challenge on CART, NPY, and leptin receptor (Ob-R) messenger ribonucleic acid (mRNA) in the nonhuman primate (NHP) hypothalamus. Ten adult female rhesus monkeys were either maintained on a regular diet or fasted for two days before euthanasia. CART, NPY, and Ob-R mRNA were measured by in situ hybridization histochemistry (ISHH). A 2-day fast decreased CART expression in the ARC, increased NPY gene expression in the supraoptic nucleus (SON) and paraventricular nucleus (PVN), and increased Ob-R expression in the ventromedial nucleus (VMN). This is the first report that fasting inhibits CART expression and stimulates Ob-R expression in monkeys. Increased NPY expression in the SON and PVN, but not the ARC of fasted monkeys also is novel. With some exceptions, our observations are confirmatory of findings in rodent studies. Similarities in the neuroendocrine responses to a metabolic challenge in monkeys and rodents support extending existing hypotheses of neuroendocrine control of energy homeostasis to primates.
Effects of Diabetes Mellitus, Chronic Renal Failure and Hemodialysis on Serum and Salivary Antioxidant Status
Authors: Ben-Zvi Ilan , Green Yaakov , Nakhoul Farid , Kanter Yoram , Nagler Rafael M.
Published: December 2006

Aim: To analyze various oxidative stress parameters in the saliva and serum of patients with chronic renal failure (CRF) and/or diabetes mellitus (DM), and to compare them in dialytic vs. pre-dialytic patients. Method: 50 consenting patients were divided into five subgroups of patients: severe CRF (dialytic) without DM, severe CRF (dialytic) with DM, mild CRF (pre-dialytic) without DM, mild CRF (pre-dialytic) with DM, and with DM but without CRF (controls). Uric acid (UA), peroxidase and total antioxidant status (TAS) were studied in both saliva and serum; superoxide dismutase (SOD) was evaluated only in saliva. Both saliva collection and serum harvesting were done simultaneously. Results: In severe-CRF patients without DM, median TAS, UA and SOD levels decreased following dialysis (54, 85, 48%, respectively), and peroxidase levels increased slightly (9%). In severe-CRF patients with DM, median TAS and SOD levels increased following dialysis (33 and 54%, respectively) while median UA and peroxidase levels decreased (68 and 10%, respectively). Conclusions: DM, CRF and hemodialysis were found to increase the oxidative stress burden in both serum and saliva. Therefore, antioxidant assessment may be used to monitor baseline oxidative status in these situations though larger randomized studies are in order.
Microembolic Signals within 24 Hours of Stroke Onset and Diffusion-Weighted MRI Abnormalities
Authors: Nakajima Makoto , Kimura Kazumi , Shimode Atsuko , Miyashita Fumio , Uchino Makoto , Naritomi Hiroaki , Minematsu Kazuo
Published: December 2006

Background: The clinical relevance of the microembolic signals (MES) detected by transcranial Doppler sonography (TCD) in acute stroke remains unclear. In a prospective study the authors analyzed the relationship between MES and the findings on diffusion-weighted magnetic resonance imaging (DWI) in acute stroke patients. Methods: We performed TCD for a period of 30 min to detect MES in patients within 24 h of stroke onset, and DWI was done within the initial 7 days. MES were assessed from Doppler waves obtained from the middle cerebral artery contralateral to the side of the neurological deficits. The acute ischemic lesions observed on DWI were classified by their diameter (small, medium or large) and by their site (cortical, superficial perforator territory, internal borderzone or deep perforator territory). Results: We obtained Doppler waves from 39 vessels in 37 patients; 2 patients had bilateral deficits. MES were detected in 12 vessels (MES-positive group) and not detected in 27 vessels (MES-negative group). No significant differences in clinical features were observed between the 2 groups. The number of small lesions was significantly higher in the MES-positive group than in the MES-negative group (p = 0.02). The numbers of cortical and superficial perforator infarcts were significantly higher in the MES-positive group than in the MES-negative group (p = 0.002 and 0.02, respectively). Conclusion: In acute ischemic stroke, MES detected by TCD in the acute phase may produce small cortical and subcortical lesions found on DWI.