bluuebunny/crossref_metadata_2025 huggingface dataset:
Osteocytes inhibit osteoclastic bone resorption through transforming growth factor‐β: Enhancement by estrogen*
Authors: Heino Terhi J. , Hentunen Teuvo A. , Väänänen H. Kalervo
Published: October 2004

Osteocytes are the most abundant cells in bone and distributed throughout the bone matrix. They are connected to the each other and to the cells on the bone surface. Thus, they may also secrete some regulatory factors controlling bone remodeling. Using a newly established osteocyte‐like cell line MLO‐Y4, we have studied the interactions between osteocytes and osteoclasts. We collected the conditioned medium (CM) from MLO‐Y4 cells, and added it into the rat osteoclast cultures. The conditioned medium had no effect on osteoclast number in 24‐h cultures, but it dramatically inhibited resorption. With 5, 10, and 20% CM, there was 25, 39, and 42% inhibition of resorption, respectively. Interestingly, the inhibitory effect was even more pronounced, when MLO‐Y4 cells were pretreated with 10−8 M 17‐β‐estradiol. With 5, 10, and 20% CM, there was 46, 51, and 58% of inhibition. When the conditioned medium was treated with neutralizing antibody against transforming growth factor‐β (TGF‐β), the inhibitory effect was abolished. This suggests that osteocytes secrete significant amounts of TGF‐β, which inhibits bone resorption and is modulated by estrogen. RT–PCR and Western blot analysis show that in MLO‐Y4 cells, the prevalent TGF‐β isoform is TGF‐β3. We conclude that osteocytes have an active, inhibitory role in the regulation of bone resorption. Our results further suggest a novel role for TGF‐β in the regulation of communication between different bone cells and suggest that at least part of the antiresorptive effect of estrogen in bone could be mediated via osteocytes. J. Cell. Biochem. 85: 185–197, 2002. © 2002 Wiley‐Liss, Inc.
Nigericin inhibits insulin‐stimulated glucose transport in 3T3‐L1 adipocytes
Authors: Chu Chih‐Ying , Kao Ying‐Shun , Fong Jim C.
Published: October 2004

We used nigericin, a K+/H+ exchanger, to test whether glucose transport in 3T3‐L1 adipocytes was modulated by changes in intracellular pH. Our results showed that nigericin increased basal but decreased insulin‐stimulated glucose uptake in a time‐ and dose‐dependent manner. Whereas the basal translocation of GLUT1 was enhanced, insulin‐stimulated GLUT4 translocation was inhibited by nigericin. On the other hand, the total amount of neither transporter protein was altered. The finding that insulin‐stimulated phosphoinositide 3‐kinase (PI 3‐kinase) activity was not affected by nigericin implies that nigericin exerted its inhibition at a step downstream of PI 3‐kinase activation. At maximal dose, nigericin rapidly lowered cytosolic pH to 6.7; however, this effect was transient and cytosolic pH was back to normal in 20 min. Removal of nigericin from the incubation medium after 20 min abolished its enhancing effect on basal but had little influence on its inhibition of insulin‐stimulated glucose transport. Moreover, lowering cytosolic pH to 6.7 with an exogenously added HCl solution had no effect on glucose transport. Taken together, it appears that nigericin may inhibit insulin‐stimulated glucose transport mainly by interfering with GLUT4 translocation, probably by a mechanism not related to changes in cytosolic pH. J. Cell. Biochem. 85: 83–91, 2002. © 2002 Wiley‐Liss, Inc.
Reduced CpG methylation is associated with transcriptional activation of the bone‐specific rat osteocalcin gene in osteoblasts*
Authors: Villagra Alejandro , Gutiérrez José , Paredes Roberto , Sierra José , Puchi Marcia , Imschenetzky Maria , van Wijnen Andre , Lian Jane , Stein Gary , Stein Janet , Montecino Martin
Published: October 2004

Chromatin remodeling of the bone‐specific rat osteocalcin (OC) gene accompanies the onset and increase in OC expression during osteoblast differentiation. In osseous cells expressing OC, the promoter region contains two nuclease hypersensitive sites that encompass the elements that regulate basal tissue‐specific and vitamin D‐enhanced OC transcription. Multiple lines of evidence indicate that DNA methylation is involved in maintaining a stable and condensed chromatin organization that represses eukaryotic transcription. Here we report that DNA methylation at the OC gene locus is associated with the condensed chromatin structure found in cells not expressing OC. In addition, we find that reduced CpG methylation of the OC gene accompanies active transcription in ROS 17/2.8 rat osteosarcoma cells. Interestingly, during differentiation of primary diploid rat osteoblasts in culture, as the OC gene becomes increasingly expressed, CpG methylation of the OC promoter is significantly reduced. Inhibition of OC transcription does not occur by a direct mechanism because in vitro methylated OC promoter DNA is still recognized by the key regulators Runx/Cbfa and the vitamin D receptor complex. Furthermore, CpG methylation affects neither basal nor vitamin D‐enhanced OC promoter activity in transient expression experiments. Together, our results indicate that DNA methylation may contribute indirectly to OC transcriptional control in osteoblasts by maintaining a highly condensed and repressed chromatin structure. J. Cell. Biochem. 85: 112–122, 2002. © 2002 Wiley‐Liss, Inc.
DNA replication and nuclear architecture
Authors: Jaunin Françoise , Fakan Stanislav
Published: October 2004

The model of in situ DNA replication provided by immunofluorescence and confocal imaging is compared with observations obtained by electron microscopic studies. Discrepancies between both types of observations call into question the replication focus as a persistent nuclear structure and as a replication entity where DNA replication takes place. Most electron microscopic analyses reveal that replication sites are confined to dispersed chromatin areas at the periphery of condensed chromatin, and the distribution of replication factors exhibits the same localization pattern. Moreover, rapid migration of newly synthesized DNA from the replication sites towards the interior of condensed chromatin regions obviously takes place during S‐phase. It implies modifications of replication domains, hardly detectable by fluorescence microscopy. The confrontation of different observations carried out at light microscopic or electron microscopic levels of resolution lead to a conclusion that a combination of in vivo fluorescence analysis with a subsequent ultrastructural investigation performed on the same cells will represent an optimal approach in future studies of nuclear functions in situ. J. Cell. Biochem. 85: 1–9, 2002. © 2002 Wiley‐Liss, Inc.
Soluble fragment of P‐cadherin adhesion protein found in human milk
Authors: Soler Alejandro Peralta , Russo Jose , Russo Irma H. , Knudsen Karen A.
Published: October 2004

Classical cadherins such as E‐ and P‐cadherin are transmembrane proteins that mediate specific cell‐to‐cell adhesion and are important to tissue development and function. Cadherin function can be modulated by various means, including proteolytic cleavage of the extracellular adhesion domain from the cells' surface, yielding large soluble fragments termed (soluble) sE‐ or sP‐cadherin. In people with certain carcinomas, sE‐cadherin can be detected at elevated levels in the serum and sometimes can serve as a prognostic marker. Soluble E‐cadherin also is found in urine of patients with bladder cancer. In addition to being present in bodily fluids of cancer patients, sE‐ and sP‐cadherin are present in the serum of healthy people, suggesting that shedding of cadherins is a normal event. Here, we report high levels of 80 kDa sP‐cadherin in human milk. In the lactating mammary gland tissue, P‐cadherin appears to be a protein secreted by epithelial cells, rather than an adhesion protein. This is in contrast to the non‐lactating mammary gland where P‐cadherin is restricted to myoepithelial cells, and is present at sites of cell–cell contact. J. Cell. Biochem. 85: 180–184, 2002. © 2002 Wiley‐Liss, Inc.
1‐Rotational Steiner triple systems over arbitrary groups
Authors: Buratti Marco
Published: October 2004

Phelps and Rosa introduced the concept of 1‐rotational Steiner triple system, that is an STS(ν) admitting an automorphism consisting of a fixed point and a single cycle of length ν − 1 [Discrete Math. 33 (1981), 57–66]. They proved that such an STS(ν) exists if and only if ν ≡ 3 or 9 (mod 24). Here, we speak of a 1‐rotational STS(ν) in a more general sense. An STS(ν) is 1‐rotational over a group G when it admits G as an automorphism group, fixing one point and acting regularly on the other points. Thus the STS(ν)'s by Phelps and Rosa are 1‐rotational over the cyclic group. We denote by 𝒜1r, 𝒞1r, 𝒬1r, 𝒢1r, the spectrum of values of ν for which there exists a 1‐rotational STS(ν) over an abelian, a cyclic, a dicyclic, and an arbitrary group, respectively. In this paper, we determine 𝒜1r and find partial answers about 𝒬1r and 𝒢1r. The smallest 1‐rotational STSs have orders 9, 19, 25 and are unique up to isomorphism. In particular, the only 1‐rotational STS(25) is over SL2(3), the special linear group of dimension 2 over Z3. © 2001 John Wiley & Sons, Inc. J Combin Designs 9: 215–226, 2001
There exists no (15,5,4) RBIBD
Authors: Kaski Petteri , Östergård Patric R. J.
Published: October 2004

An (n, M, d)q code is a q‐ary code of length n, cardinality M, and minimum distance d. We show that there exists no (15,5,4) resolvable balanced incomplete block design (RBIBD) by showing that there exists no (equidistant) (14,15,10)3 code. This is accomplished by an exhaustive computer search using an orderly algorithm combined with a maximum clique algorithm. © 2001 John Wiley & Sons, Inc. J Combin Designs 9: 227–232, 2001
A hole‐size bound for incomplete t‐wise balanced designs
Authors: Kreher Donald L. , Rees Rolf S.
Published: October 2004

An incomplete t‐wise balanced design of index λ is a triple (X,H,ℬ︁) where X is a υ–element set, H is a subset of X called the hole, and B is a collection of subsets of X called blocks, such that, every t‐element subset of X is either in H or in exactly λ blocks, but not both. If H is a hole in an incomplete t‐wise balanced design of order υ and index λ, then |H| ≤ υ/2 if t is odd and |H| ≤ (υ − 1)/2 if t is even. In particular, this result establishes the validity of Kramer's conjecture that the maximal size of a block in a Steiner t‐wise balanced design is at most υ/2 if t is odd and at most (υ−1)/2 when t is even. © 2001 John Wiley & Sons, Inc. J Combin Designs 9: 269–284, 2001
The Neuropsychological Impairment Scale—Senior: A procedure for evaluating awareness disturbance in geriatric patients
Authors: O'Donnell William E.
Published: October 2004

The Neuropsychological Impairment Scale—Senior (NIS‐S) is a 30‐item, orally administered questionnaire for geriatric patients about symptoms of cognitive impairment, affective disturbance, and defensiveness. Clinical and nonclinical norms are described along with validity and reliability information. A procedure is presented for comparing a patient's self‐reported cognitive impairment with scores on performance testing, which gauges whether the patient is over‐ or under‐reporting cognitive deficits. © 2001 John Wiley & Sons, Inc. J Clin Psychol 57: 423–427, 2001.
The responses of dissociative patients on the thematic apperception test
Authors: Pica Michael , Beere Donald , Lovinger Sophie , Dush David
Published: October 2004

This study compared the responses of dissociative inpatients and general inpatient psychiatric controls on the Thematic Apperception Test (TAT; Murray, 1943). We found the stories of dissociative participants to be characterized by a greater interpersonal distance and more trauma and dissociation responses than those of the controls. No significant differences were found regarding total number of emotional references, although references to positive emotions were almost nonexistent for the dissociative group. A post hoc analysis of the data found the testing behaviors of dissociative participants to be characterized by switching, trance states, intrainterview amnesias, and affectively loaded card rejections. Questions were raised regarding the relevancy of the findings to clinical practice and how they might explain some of the controversies surrounding the diagnosis of dissociative identity disorder (DID). © 2001 John Wiley & Sons, Inc. J Clin Psychol 57: 847–864, 2001.
Prediction of dose–response relations based on patient characteristics
Authors: Lutz Wolfgang , Lowry Jenny , Kopta S. Mark , Einstein Danielle A. , Howard Kenneth I.
Published: October 2004

The recent discussion of evidence‐based, adaptive treatment planning highlights the need for models for the prediction of courses of treatment response. We combine a dose–response model with growth curve modeling to determine dose–response relations for well‐being, symptoms, and functioning. Hierarchical linear modeling was used to model each patient's expected course of improvement. The resulting predictions were cross‐validated on two samples of psychotherapy outpatients. The results give further empirical support for the dose–response model and the phase model of psychotherapy as well as for the usefulness of patient treatment response profiling for individual treatment management. © 2001 John Wiley & Sons, Inc. J Clin Psychol 57: 889–900, 2001.
Power Doppler sonography of hepatocellular carcinomas with portal‐vein blood supply
Authors: Tohara Keiji , Sakaguchi Seigo , Miyajima Yasushi , Yao Tsuneyoshi , Ono Hiroyuki
Published: October 2004

We report the power Doppler imaging (PDI) findings in 2 cases of hepatocellular carcinoma (HCC) with a portal‐vein blood supply. Gray‐scale sonography in both cases showed a well‐circumscribed nodule, hypoechoic in case 1 and hyperechoic in case 2. PDI revealed an afferent tumor vessel with constant flow in both nodules, and CT during arterial portography demonstrated a portal‐vein supply to both nodules. The nodules were diagnosed by percutaneous core biopsies as highly differentiated HCC. We also examined with PDI another 64 patients with 76 HCC nodules, and none of the nodules showed a constant‐flow afferent tumor vessel. The presence of a constant‐flow afferent tumor vessel indicates a supplying portal vein; but this is not diagnostic of HCC, and biopsy remains necessary to establish a final diagnosis. © 2001 John Wiley & Sons, Inc. J Clin Ultrasound 29:290–293, 2001.
Partially reversed intrasplenic venous blood flow detected by color Doppler sonography in two patients with hematologic diseases and splenomegaly
Authors: Restrepo‐Schäfer Isabel , Wollenberg Birgit , Riera‐Knorrenschild Jordi , Görg Christian
Published: October 2004

We report on 2 patients with hematologic diseases (1 follicular lymphoma and 1 myeloproliferative syndrome) and splenomegaly who had partial intrasplenic portosystemic shunting demonstrated by color Doppler sonography. Intrasplenic venous blood flow was in the normal direction at the hilum of the spleen but in a reversed direction at the periphery of the spleen. This type of reversed intrasplenic flow pattern results in portosystemic shunting and might be detected more frequently when careful color Doppler mapping of the entire splenic parenchyma is performed in patients with portal hypertension. The clinical significance of this phenomenon, however, is still unclear. © 2001 John Wiley & Sons, Inc. J Clin Ultrasound 29:294–297, 2001.
Endorectal sonographic appearances of rectal MALT lymphoma, its response to therapy, and local recurrence
Authors: Gavioli Margherita , Bagni Alberto , Santacroce Giovanni , Piccagli Italo , Natalini Gianni
Published: October 2004

Endorectal sonography may significantly help to evaluate rectal lymphoma. We report the sonographic findings in a case of rectal non‐Hodgkin's MALT (mucosa‐associated lymphoid tissue) lymphoma, including the monitoring of response to therapy and confirmation of recurrence, in a 45‐year‐old man. On endorectal sonography of the rectal wall, the mucosa was markedly thickened to 1.1 cm and was diffusely hypoechoic and risen into multiple polypoid folds. The submucosa and muscularis propria appeared normal. Multiple lymph nodes were visualized in the perirectal fat; they were homogeneously hypoechoic, were round or oval, and ranged from 1.0 cm to 2.6 cm. Endoscopic biopsies revealed a grade I non‐Hodgkin's MALT lymphoma. Following chemotherapy, endorectal sonography showed that the surface of the rectal mucosa had a smoother appearance and near‐normal thickness, but lymph nodes, although smaller, remained visible in the perirectal fat. Four months later, endorectal sonography demonstrated a local relapse of disease, with significant thickening of the rectal mucosa and multiple lymph nodes visible in the perirectal fat. Following high‐dose chemotherapy for the recurrence, endorectal sonography demonstrated a near‐normal appearance of the rectal mucosa. © 2001 John Wiley & Sons, Inc. J Clin Ultrasound 29:401–405, 2001.
Fungal brain abscesses in neonates: Sonographic appearances and corresponding histopathologic findings
Authors: Marcinkowski Monika , Bauer Karl , Stoltenburg‐Didinger Gisela , Versmold Hans
Published: October 2004

Extremely preterm neonates and neonates with predisposing conditions such as congenital or acquired immunodeficiency are at high risk for systemic fungal infection. Abscess formation in the brain is a severe complication that occurs in 70% of neonates with systemic fungal infection. Cerebral sonography can be used to diagnose abscesses in the brain in these patients. We report 2 sonographic presentations of fungal brain abscesses in neonates confirmed by postmortem histopathologic examination. The first patient, an extremely preterm neonate of 23 weeks' gestation with a systemic Candida albicans infection, had multiple small, round, hypoechoic lesions with echogenic rims in both brain hemispheres. The second patient, a term neonate with disseminated aspergillosis and DiGeorge syndrome, had a few large echogenic areas in the right periventricular region. Brain imaging should be considered in the diagnostic workup in neonates with suspected systemic fungal infection. Cerebral involvement can be diagnosed at the bedside with cerebral sonography. © 2001 John Wiley & Sons, Inc. J Clin Ultrasound 29:417–421, 2001.
Lymphangiogenesis and lymphangiodysplasia: From molecular to clinical lymphology
Authors: Witte Marlys H. , Bernas Michael J. , Martin Clifford P. , Witte Charles L.
Published: October 2004

The lymph vascular system parallels the blood vasculature and as one of its key functions returns liquid and solutes to the bloodstream, including macromolecules that have escaped from blood capillaries and entered the interstitium. In conjunction with interspersed lymph nodes and lymphoid organs, the lymphatic vasculature also acts as a conduit for trafficking immune cell populations. Echoing the explosion of knowledge about blood vessel angiogenesis (properly termed “hemangiogenesis”), the past two decades have also witnessed a series of significant, yet less‐noticed discoveries bearing on “lymphangiogenesis,” along with delineation of the spectrum of lymphedema–angiodysplasia syndromes. Failure of lymph transport promotes a brawny proteinaceous edema of the affected limb, organ, or serous space that is disfiguring, disabling, and on occasion even life‐threatening. Key members of the vascular endothelial growth factor (VEGF) and angiopoietin families of vascular growth factors (and their corresponding tyrosine kinase endothelial receptors) have been identified which preferentially influence lymphatic growth and, when manipulated in genetically engineered murine models, produce aberrant “lymphatic phenotypes.” Moreover, mutations in VEGF receptor and forkhead family developmental genes have now been linked and implicated in the pathogenesis of two familial lymphedema–angiodysplasia syndromes. Thus, recent advances in “molecular lymphology” are elucidating the poorly understood development, physiology, and pathophysiology of the neglected lymphatic vasculature. In combination with fresh insights and refined tools in “clinical lymphology,” these advances should lead not only to earlier detection and more rational classification of lymphatic disease but also to better therapeutic approaches, including designer drugs for lymphangiostimulation and lymphangioinhibition and gene therapy to modulate lymphatic growth. Microsc. Res. Tech. 55:122–145, 2001. © 2001 Wiley‐Liss, Inc.
Muscle regeneration in holothurians
Authors: Dolmatov Igor Yu. , Ginanova Talia T.
Published: October 2004

The muscle system of holothurians includes visceral (coelomic epithelium) and somatic (longitudinal muscle bands, retractors of aquapharyngeal complex) musculature. Visceral musculature regeneration is achieved by the transformation of myoepithelial cells via their dedifferentiation, migration, proliferation, and redifferentiation. During somatic muscle regeneration the new muscle bundles are formed due to dedifferentiation, migration, and immersion of the coelomic epithelial cells into the connective tissue. While submerging, the epithelial cells transform into myocytes and begin to produce myofibrils in their cytoplasm. Concomitantly, a basal lamina is formed around the group of myogenic cells, separating them from the surrounding extracellular matrix. The myohistogenesis is accompanied by a conspicuous DNA‐synthetic activity. Proliferation is insignificant and seems to be of no essential importance for muscle regeneration. The synthesis of DNA followed by no cytokinesis results in an increase in the amount of DNA of myocyte nuclei. Microsc. Res. Tech. 55:452–463, 2001. © 2001 Wiley‐Liss, Inc.
Small regular graphs having the same path layer matrix*
Authors: Yuansheng Yang , Jianhua Lin , Chunli Wang
Published: October 2004

The path layer matrix of graph G contains quantitative information about all paths in G. The entry (i,j) in this matrix is the number of simple paths in G having initial vertex i and length j. Some new upper bounds for r‐regular graphs with the same path layer matrix are presented for r=4, 5, 6. © 2002 Wiley Periodicals, Inc. J Graph Theory 39: 219–221, 2002; DOI 10.1002/jgt.10005
Girth and fractional chromatic number of planar graphs
Authors: Pirnazar Amir , Ullman Daniel H.
Published: October 2004

In 1959, even before the Four‐Color Theorem was proved, Grötzsch showed that planar graphs with girth at least 4 have chromatic number at the most 3. We examine the fractional analogue of this theorem and its generalizations. For any fixed girth, we ask for the largest possible fractional chromatic number of a planar graph with that girth, and we provide upper and lower bounds for this quantity. © 2002 Wiley Periodicals, Inc. J Graph Theory 39: 201–217, 2002; DOI 10.1002/jgt10024
Introduction to the Rhizocephala (Crustacea: Cirripedia)
Authors: Walker Graham
Published: October 2004

Knowledge about the Rhizocephala (parasitic barnacles) has increased exponentially over the past two decades. This introduction broadly reviews this progress, touching on rhizocephalan morphology, life‐cycles, larval biology, and the effects on the crustacean hosts. Members representing both orders, the Kentrogonida and Akentrogonida, are discussed. The recent discovery regarding the vermigon stage further highlights the intricacy of the kentrogonid life‐cycle. Because rhizocephalans are found in most world regions, occurring on their respective hosts from the deep ocean to freshwater, their importance is now being recognized. J. Morphol. 249:1–8, 2001. © 2001 Wiley‐Liss, Inc.
Ephrins stimulate neurite outgrowth during early cortical neurogenesis
Authors: Zhou Xiaofeng , Suh Junghyup , Cerretti Douglas P. , Zhou Renping , DiCicco‐Bloom Emanuel
Published: October 2004

The Eph receptor ligands, the ephrins, are membrane‐bound molecules that play important roles in establishing intercellular communication after neurogenesis by regulating cell migration, axon pathfinding, and topographic mapping. In diverse systems, such as embryonic day 17.5 (E17.5) hippocampal and cortical neurons, repulsive/inhibitory mechanisms underlie these cellular effects. However, although ligand/receptor expression occurs far earlier, during brain neurogenesis, little is known about potential ephrin functions in initial process outgrowth. We have examined ligand/receptor expression in E13.5 cortex in vivo and in culture, using alkaline phosphatase (AP)‐conjugated reagents and RNase protection assay. B ephrins are highly expressed, including B1, B2, and B3, whereas A ephrins exhibit low expression levels. In contrast, the Eph receptors demonstrate an opposite pattern, exhibiting high levels of Eph A3, A4, and A5 mRNA transcripts and low levels of the B‐class receptors. To examine effects on neurite outgrowth, soluble ephrins were incubated with antihuman IgG antibody, producing oligomeric agonist complexes, and dried onto culture dishes. Unexpectedly, both ephrin A and B complexes increased process outgrowth: Seventy to eighty percent of neuronal precursors exhibited long neurites on ephrins, whereas only 5–10% of cells had neurites on IgG control substrates, indicating that ephrins stimulated neuritogenesis by early cortical neurons. These observations suggest that ephrin ligand/receptor systems play ontogenetic roles not previously considered, activating mechanisms other than cellular repulsion. Ephrin systems may induce initial process elaboration by early cortical neurons that is restricted at later stages by well‐characterized repulsive signaling mechanisms. J Neurosci. Res. 66:1054–1063, 2001. © 2001 Wiley‐Liss, Inc.
Distribution and origin of corticotropin‐releasing factor‐immunoreactive axons in the female rat lumbosacral spinal cord
Authors: Puder Barbara A. , Papka Raymond E.
Published: October 2004

Corticotropin‐releasing factor (CRF) is a neuropeptide traditionally known for its hormonal role in the hypothalamic/pituitary/adrenal stress axis. However, CRF has been reported in axons in sites that may be considered outside of the direct stress axis, e.g., in axons in the lumbosacral spinal cord associated with the micturition response. Whether any of these CRF‐immunoreactive axons interacts with uterine‐related preganglionic autonomic neurons or projection neurons in the lumbosacral spinal cord is unknown. Thus, immunohistochemistry and retrograde tracing were employed to determine the presence, distribution, and origin of CRF‐immunoreactive axons in the L6/S1 spinal cord of the female rat and to ascertain whether these axons are associated with uterine‐related neurons. CRF‐immunoreactive axons were present in the dorsal horn, medial and lateral collateral pathways, dorsal intermediate gray, laminae VlI and X, and sacral parasympathetic nucleus of the spinal cord. Nitric oxide‐synthesizing, i.e., NADPH‐d‐positive neurons and pseudorabies virus labeled uterine‐related neurons were in the sacral parasympathetic nucleus and were closely apposed by CRF‐immunoreactive axons. Injection of retrograde tracers (fluorogold or fast blue) into the L6/S1 spinal cord labeled neurons in the hypothalamic paraventricular nucleus and pontine Barrington's nucleus, and some of these neurons were immunoreactive for CRF. This study demonstrates that CRF‐immunoreactive axons are present in the L6/S1 spinal cord of the female rat in areas associated with sensory and autonomic processing. Some of these axons originate from the paraventricular nucleus and Barrington's nucleus and are adjacent to uterine‐related neurons. These results indicate that CRF may influence neural activity related to the female reproductive system. J Neurosci. Res. 66:1217–1225, 2001. © 2001 Wiley‐Liss, Inc.
Huperzine A attenuates amyloid β‐peptide fragment 25‐35‐induced apoptosis in rat cortical neurons via inhibiting reactive oxygen species formation and caspase‐3 activation
Authors: Xiao Xiao Qiu , Zhang Hai Yan , Tang Xi Can
Published: October 2004

Huperzine A, a novel Lycopodium alkaloid originally discovered in the Chinese herb Qian Ceng Ta (Huperzia serrata), is a reversible, potent, and selective acetylcholinesterase (AChE) inhibitor and has been extensively used for the treatment of Alzheimer's disease (AD) in China. The present studies were designed to investigate effects of huperzine A on amyloid β‐peptide fragment 25‐35 (Aβ25‐35)‐induced neuronal apoptosis and potential mechanisms in primary cultured rat cortical neurons. After exposure of the cells to Aβ25‐35 (20 μM), apoptotic cell death was observed as evidenced by a significant decrease in cell viability, alteration of neuronal morphology, and DNA fragmentation. Pretreatment of the cells with huperzine A (0.01–10 μM) prior to Aβ25‐35 exposure significantly elevated the cell survival and reduced Aβ25‐35‐induced nuclei fragmentation. Reactive oxygen species (ROS)‐based fluorescence, caspase‐3‐like fluorogenic cleavage, and Western blot analysis demonstrated that huperzine A reduced Aβ25‐35‐induced ROS formation in a dose‐dependent manner, and 1 μM of huperzine A attenuated Aβ25‐35‐induced caspase‐3 activity at 6, 12, 24, and 48 hr posttreatment. Our results provide the first direct evidence that huperzine A protects neurons against Aβ25‐35‐induced apoptosis via the inhibition of ROS formation and caspase‐3 activity. © 2002 Wiley‐Liss, Inc.
Regulation of striatal neuropeptide mRNAs: Effects of the 5‐HT2 antagonist SR46349B in adult rats with a neonatal 6‐hydroxydopamine lesion
Authors: Durand C. , Mathieu‐Kia A.M. , Besson M.‐J. , Penit‐Soria J.
Published: October 2004

The intrastriatal injection of 6‐hydroxydopamine (6‐OHDA) in newborn rats produces a marked striatal dopamine (DA) depletion, accompanied by a serotonin (5‐HT) hyperinnervation and an up‐regulation of 5‐HT receptors. The aim of the present study was to investigate whether the increase in 5‐HT2 receptors could compensate for some of the DA lesion‐induced effects, such as the increase in striatal preproenkephalin (PPE) and the decrease in preprotachykinin A (PPT‐A) mRNA levels. Three months after the DA lesion, the effect of the selective 5‐HT2 antagonist SR46349B was investigated by a subacute treatment (10 mg/kg, IP, twice per day for 3.5 days). In sham‐operated rats, the blockade of 5‐HT2 receptors decreased PPE mRNA levels in the striatum and, by contrast, had no effect on PPT‐A mRNA levels. In rats with a unilateral neonatal DA lesion, SR46349B had no more effect on PPE mRNA levels in the intact striatum and was unable to modify the lesion induced‐increase in PPE mRNA. The decrease in PPT‐A mRNA levels induced by the neonatal DA lesion was not changed after SR46349B treatment in the posterior part of the lesioned striatum. Our results suggest that SR46349B indirectly decreases PPE mRNA levels in striatopallidal neurons in intact animals through a desinhibition of DA neuron activity. This is further evidenced by the lack of PPE mRNA changes in the DA lesioned striatum despite the up‐regulation of 5‐HT2 receptor transmission induced in this model. Finally, the absence of any effect of 5‐HT2 antagonist on the expression of PPT‐A mRNA in intact animals is discussed. The precise role of 5‐HT2 receptor on PPT‐A mRNA biosynthesis after a neonatal lesion should be clarified by further experiments using 5‐HT2 agonists. © 2002 Wiley‐Liss, Inc.
Abnormal reinnervation of skeletal muscle in a tenascin‐C‐deficient mouse
Authors: Cifuentes‐Diaz C. , Faille L. , Goudou D. , Schachner M. , Rieger F. , Angaut‐Petit D.
Published: October 2004

The possible involvement of tenascin‐C in the reinnervation of a skeletal muscle was investigated in the tenascin‐C‐deficient mouse (T–/–) produced by Saga et al. (1992; Genes Dev 6:1821–1831). The pattern of reinnervation, observed after denervation of the triangularis sterni muscle, differs in T–/– and wild‐type muscles in several traits. Axonal growth and stability of terminal arbors are impaired in the T–/– muscle: Some axons in mutant muscles grow beyond their original targets and reinnervate other synaptic sites, which may become dually innervated. In contrast to wild type, polyinnervation increases with time after denervation in T–/– muscles and is still present 7 months after nerve crush. The expression of a tenascin‐C mRNA product disappears between 1 and 2 months after nerve crush. Of interest is that this transcriptional regulation in T–/– muscles occurs when major alterations in the morphology of regenerating endings become obvious. These observations strongly implicate tenascin‐C in the formation, maturation, and stabilization of the neuromuscular junction. © 2002 Wiley‐Liss, Inc.
Differential regulation of GDNF, neurturin, and their receptors in primary cultures of rat glial cells
Authors: Rémy Séverine , Naveilhan Philippe , Brachet Philippe , Neveu Isabelle
Published: October 2004

Glial cell line‐derived neurotrophic factor (GDNF) and neurturin (NTN) bind to GFRα‐1 and GFRα‐2 receptors, respectively, and their neurotrophic activity is mediated by the tyrosine kinase receptor, Ret. All these molecules were found to be expressed in primary cultures of rat glial cells, which were largely composed of astrocytes and maintained in serum‐free medium. Although GDNF, NTN and Ret mRNA levels were at the limit of detection, RNase protection assays revealed relatively high amounts of GFRα‐1 and GFRα transcripts. To characterize signals controlling their expression, glial cells were exposed to serum or treated with hormones acting through nuclear receptors and by activators of the cAMP or protein kinase C (PKC)‐dependent pathways. Retinoic acid or 1,25‐dihydroxyvitamin D3 appeared ineffective. In contrast, the 5‐fold increase in GFRα‐2 mRNA after 24 hr of treatment with 10−10 M of tri‐iodothyronine, suggests a physiological role of thyroid hormone in the regulation of this receptor in vivo. The serum induced a 7‐fold increase in GFRα‐1 mRNA levels. These changes may be mediated by the cAMP or PKC pathways because both forskolin and TPA up‐regulated the GFRα‐1 gene. Interestingly, only TPA led to a coordinated increase in the levels of GDNF, GFRα‐1 and GFRα‐2 mRNAs. On the other hand, NTN transcripts remained constant, irrespective of the culture conditions. Taken together, these results indicate that GDNF family ligands and their receptors are regulated in glial cells by common or independent transductional pathways, which could modulate their specific expression during brain development or in the case of trauma. J. Neurosci. Res. 64:242–251, 2001. © 2001 Wiley‐Liss, Inc.
Differential regulation of telomerase activity and TERT expression during brain development in mice
Authors: Klapper Wolfram , Shin Thuzar , Mattson Mark P.
Published: October 2004

Telomerase is an enzyme activity consisting of a reverse transcriptase called TERT and an RNA component that adds repeats of a DNA sequence (TTAGGG) to the ends of chromosomes, thereby preventing their shortening. Associations between telomerase activity and proliferation and differentiation of neural tumor cells and neural stem cells have been reported, but the role of telomerase in brain development is unknown. We now report analyses of telomerase activity, TERT mRNA levels and levels of mRNAs encoding the telomere‐associated proteins TRF1 and TRF2 in three different brain regions (brainstem, hippocampus and cerebral cortex) and the eye of mice at increasing developmental time points. Telomerase activity is high in the brain at embryonic day 13 (E13), declines markedly between E13 and E18, remains at a low level until postnatal day 3 (P3) and becomes undetectable by P10. Surprisingly, the temporal pattern of change in telomerase activity is not paralleled by a decrease in levels of TERT mRNA that remain elevated from E13 to P5 (with fluctuations during this time window that vary among brain regions), and then decrease to a lower level that is maintained into adulthood. TRF1 and TRF2 mRNA levels are relatively constant throughout brain development. Our data are consistent with a role for telomerase activity in proliferation of neural progenitor cells, and further suggest that TERT may play roles in neuronal differentiation and survival. The dissociation between TERT expression and telomerase activity is a novel finding that suggests biological functions for TERT in addition to telomere maintenance. J. Neurosci. Res. 64:252–260, 2001. Published 2001 Wiley‐Liss, Inc.
Effect of osmotic stress on potassium accumulation around glial cells and extracellular space volume in rat spinal cord slices
Authors: Vargová Lýdia , Chvátal Alexandr , Anděrová Miroslava , Kubinová Šárka , Žiak Drahomír , Syková Eva
Published: October 2004

In rat brain and spinal cord slices, the local extracellular accumulation of K+, as indicated by K+ tail currents (Itail) after a depolarization step, is greater in the vicinity of oligodendrocytes than that of astrocytes. It has been suggested that this may reflect a smaller extracellular space (ECS) around oligodendrocytes compared to astrocytes [Chvátal et al. [1997] J. Neurosci. Res. 49:98–106; [1999] J. Neurosci. Res. 56:493–505). We therefore compared the effect of osmotic stress in spinal cord slices from 5–11‐day‐old rats on the changes in reversal potentials (Vrev) of Itail measured by the whole‐cell patch‐clamp technique and the changes in ECS volume measured by the real‐time iontophoretic method. Cell swelling induced by a 20 min perfusion of hypoosmotic solution (200 mmol/kg) decreased the ECS volume fraction from 0.21 ± 0.01 to 0.15 ± 0.02, i.e., by 29%. As calculated from Vrev of Itail using the Nernst equation, a depolarizing prepulse increased [K+]e around astrocytes from 11.0 to 44.7 mM, i.e., by 306%, and around oligodendrocytes from 26.1 to 54.9 mM, i.e., by 110%. The ECS volume fraction decrease had the same time course as the changes in Vrev of Itail. Cell shrinkage in hyperosmotic solution (400 mmol/kg) increased ECS volume fraction from 0.24 ± 0.02 to 0.32 ± 0.02, i.e., by 33%. It had no effect on [K+]e evoked by a depolarizing prepulse in astrocytes, whereas in oligodendrocytes [K+]e rapidly decreased from 52 to 26 mM, i.e., by 50%. The increase in ECS volume was slower than the changes in [K+]e. These data demonstrate that hypoosmotic solution has a larger effect on the ECS volume around astrocytes than around oligodendrocytes and that hyperosmotic solution affects the ECS volume around oligodendrocytes only. This indicates that increased K+ accumulation in the vicinity of oligodendrocytes could be due to a restricted ECS. Oligodendrocytes in the CNS are therefore most likely surrounded by clusters of “compacted” ECS, which may selectively affect the diffusion of neuroactive substances in specific areas and directions and facilitate spatial K+ buffering. J. Neurosci. Res. 65:129–138, 2001. © 2001 Wiley‐Liss, Inc.
Guanosine‐induced increase in free cytosolic calcium concentration in mouse astrocytes in primary cultures: Does it act on an A3 adenosine receptor?
Authors: Chen Ye , Rathbone Michel P. , Hertz Leif
Published: October 2004

Purinergic receptors play an important role in the regulation of free cytosolic calcium concentration ([Ca2+]i) in astrocytes. In the present study, 10 μM adenosine caused an increase in [Ca2+]i in 85% of the cultures studied, i.e., primary cultures of mouse astrocytes, differentiated by culturing in the presence of dibutyryl cyclic AMP. Antagonist sensitivity and rapid desensitization suggested that it did so by acting on A3 receptors. Another biologically important purine, guanosine, also caused an increase in astrocytic [Ca2+]i (at concentrations of 0.1–100 μM). Although this response did not show the same rapid desensitization as the response to adenosine, it may also have been exerted on an A3 receptor. It supports this idea that inosine also caused an increase in [Ca2+]i, because inosine is known to activate A3 receptors in mast cells and structurally is even more closely related to guanosine than is adenosine. J. Neurosci. Res. 65:184–189, 2001. © 2001 Wiley‐Liss, Inc.
Biologically active sequence (KDI) mediates the neurite outgrowth function of the gamma‐1 chain of laminin‐1
Authors: Liesi Päivi , Laatikainen Timo , Wright Jerry M.
Published: October 2004

A neurite outgrowth domain of the γ1‐chain of laminin‐1 (RDIAEIIKDI) promotes axon guidance of rat hippocampal neurons, regulates the nuclear movement phase of neuronal migration, and binds to the cellular prion protein (Liesi et al. [1995] J. Neurosci. Res. 134:447–486; Matsuzawa et al. [1998] J. Neurosci. Res. 53:114–124; Graner et al. [2000] Brain Res. Mol. Brain Res. 76:85–92). Using electrophysiology and neuronal culture experiments, we show that this 10 amino acid peptide or its smaller domains induces potassium currents in primary central neurons. Both these currents and the neurotoxicity of high concentrations of the 10 amino acid peptide antigen are prevented by pertussis toxin. The smallest peptide domain capable of inducing both potassium currents and promoting neurite outgrowth of human spinal cord neurons is a tri‐peptide KDI. Our results indicate that KDI may be the biologically active domain of the γ1 laminin, capable of modulating electrical activity and survival of central neurons via a G‐protein coupled mechanism. These results expand the wide variety of functions already reported for the members of the laminin‐gene family. They suggest that biologically active peptide domains of the γ1 laminin may provide tools to promote neuronal regeneration after injuries and to enhance neuronal survival during aging and neuronal degeneration. J Neurosci. Res. 66:1047–1053, 2001. © 2001 Wiley‐Liss, Inc.
Distribution of divalent metal transporter 1 and metal transport protein 1 in the normal and Belgrade rat
Authors: Burdo J.R. , Menzies S.L. , Simpson I.A. , Garrick L.M. , Garrick M.D. , Dolan K.G. , Haile D.J. , Beard J.L. , Connor J.R.
Published: October 2004

Iron accumulation in the brain occurs in a number of neurodegenerative diseases. Two new iron transport proteins have been identified that may help elucidate the mechanism of abnormal iron accumulation. The Divalent Metal Transporter 1 (DMT1), is responsible for iron uptake from the gut and transport from endosomes. The Metal Transport Protein 1 (MTP1) promotes iron export. In this study we determined the cellular and regional expression of these two transporters in the brains of normal adult and Belgrade rats. Belgrade rats have a defect in DMT1 that is associated with lower levels of iron in the brain. In the normal rat, DMT1 expression is highest in neurons in the striatum, cerebellum, thalamus, ependymal cells lining the third ventricle, and vascular cells throughout the brain. The staining in the ependymal cells and endothelial cells suggests that DMT1 has an important role in iron transport into the brain. In Belgrade rats, there is generalized decrease in immunodetectable DMT1 compared to normal rats except in the ependymal cells. This decrease in immunoreactivity, however, was absent on immunoblots. The immunoblot analysis indicates that this protein did not upregulate to compensate for the chronic defect in iron transport. MTP1 staining is found in most brain regions. MTP1 expression in the brain is robust in pyramidal neurons of the cerebral cortex but is not detected in the vascular endothelial cells and ependymal cells. MTP1 staining in Belgrade rats was decreased compared to normal, but similar to DMT1 this decrease was not corroborated by immunoblotting. These results indicate that DMT1 and MTP1 are involved in brain iron transport and this involvement is regionally and cellularly specific. J Neurosci. Res. 66:1198–1207, 2001. © 2001 Wiley‐Liss, Inc.
Iron and the role of Coxiella burnetii in heart disease
Authors: Weinberg Eugene D.
Published: October 2004

Chronic infection of iron‐loaded coronary arteries by Chlamydia pneumoniae has been proposed as a heart disease risk factor. Another bacterial pathogen associated with increased risk for development of atherosclerosis is Coxiella burnetii. Like Chl. pneumoniae, C. burnetii is inhaled, multiplies in alveolar macrophages, subsequently becomes associated with arterial disease, and is enhanced by iron loading. J. Trace Elem. Exp. Med. 14:409–410, 2001. © 2001 Wiley‐Liss, Inc.
Nanosecond, high‐intensity pulsed laser ablation of myocardium tissue at the ultraviolet, visible, and near‐infrared wavelengths: In‐vitro study
Authors: Sato Shunichi , Ogura Makoto , Ishihara Miya , Kawauchi Satoko , Arai Tsunenori , Matsui Takemi , Kurita Akira , Obara Minoru , Kikuchi Makoto , Ashida Hiroshi
Published: October 2004

Background and Objective

A large number of clinical trials of transmyocardial laser revascularization (TMLR) have been conducted to treat severe ischemic heart diseases. A variety of laser sources have been used or tested for this treatment, however, no comprehensive study has been performed to reveal the mechanism and the optimum laser irradiation condition for the myocardium tissue ablation. There have been reported limited experimental data of the high‐intensity pulsed laser ablation of myocardium tissues.

Study Design/Materials and Methods

A 1064‐nm Q‐switched Nd:YAG laser and its 2nd (532 nm), 3rd (355 nm), and 4th (266 nm) harmonics were used for ablation experiments. At each wavelength, 25 laser pulses irradiated the porcine myocardium tissue samples at a constant laser intensity (peak laser power divided by laser spot area) of ∼2 GW/cm2 and the ablation depths were measured. During ablation, laser‐induced optical and acoustic emissions were measured to investigate the ablation mechanism at each laser wavelength. For the ablated tissues, histological observation was made with a polarization optical microscope.

Results

It was shown that the ablation efficiency did not directly depend on the linear absorption coefficient of the tissue; the ablation depth was maximized at 355 and 1064 nm, and minimized at 532 nm. Strong laser‐induced optical and acoustic emissions were observed for the 266‐ and 1064‐nm laser irradiations. The histology showed that thermal denaturation of the tissue near the ablation walls decreased with decreasing wavelength for 266, 355, and 532 nm, but it was limited for 1064 nm.

Conclusion

At the laser intensity of ∼2 GW/cm2, ablation characteristics were drastically changed for the different laser wavelengths. The results indicated that for 266, 355, and 532 nm, the tissue removal was achieved mainly through a photothermal process, but for 266 nm the intense laser‐induced plasma formation would result in a reduced laser energy coupling to the tissue. For 1064 nm, a photodisruption was most probable as a dominant tissue removal process. Because of the high ablation rate and limited thermal denaturation, the 355‐ and 1064‐nm lasers could be potential laser sources for TMLR, although further investigation is needed to discuss the clinical issues. Lasers Surg. Med. 29:464–473, 2001. © 2001 Wiley‐Liss, Inc.
Laser soft palate “stiffening”: An alternative to uvulopalatopharyngoplasty*
Authors: Wang Zhi , Rebeiz Elie E. , Shapshay Stanley M.
Published: October 2004

Background and Objective

Habitual snoring is best treated surgically, with uvulopalatophayngoplasty, but the standard “cutting” procedure poses potential morbidity. In this study, a new approach, stiffening of the soft palate with a low energy laser, was studied as a less invasive alternative.

Study Design

In an in vitro study, five fresh palates of canines were used to see acute thermal effect on the palatal tissue after laser treatment. The same laser irradiation was conducted in three living canines to observe a delayed response of the palate to the laser.

Materials and Methods

With the use of a non‐contact 1.44 μm Nd:YAG laser at 0.1 J and 20 Hz, the mucosa on the marginal area of the soft palate was irradiated for 2–3 minutes.

Following Outcome were Measured

Acute shrinkage of the soft palate and thermal effect on the mucosa, and delayed stiffening and elevation of the palate after laser irradiation.

Results

There was immediate shrinkage of the palate of about 3.0 mm in the in vitro study. In the in vivo study, a delayed palatal stiffening with 6.0–7.0 mm elevated palatal arch was found at 5 weeks. There was no morbidity after the treatment.

Conclusion

Laser stiffening of soft palate is simple, safe and effective for reduction of length and fluttering of the soft palate in the canine model. Clinical studies are warranted to evaluate its efficacy as an office treatment for snoring. Lasers Surg. Med. 30:40–43, 2002. © 2002 Wiley‐Liss, Inc.