bluuebunny/crossref_metadata_2025 huggingface dataset:
Astrocyte cultures from conditional connexin43‐deficient mice
Authors: Theis Martin , Speidel Dina , Willecke Klaus
Published: March 2004

Connexin43 (Cx43) mainly provides the molecular basis for astrocytic gap junctions. Interastrocytic coupling is thought to mediate extracellular ion homeostasis, long‐range signaling, and neuroprotection in the brain. Cx43 has been implicated in astrocytic growth control and is also expressed in other cell types in the brain, such as leptomeningeal and vascular cells. Cx43 function has been studied in astrocyte cultures of Cx43‐deficient mice, which lack Cx43 in all cell types. We have generated conditionally deficient mice with an astrocyte‐directed inactivation of Cx43, which leaves expression in other cell types unaffected. Other connexins have been detected in astrocytes. For the study of astrocytes lacking Cx45 and Cx26 in vitro, which deficiencies are embryonic lethal, conditionally deficient astrocyte cultures are essential. In the present study, we describe the developmental kinetics of Cx43 inactivation and loss of intercellular communication in astrocyte cultures derived from conditional Cx43‐deficient mice. Conditional ablation of Cx43 is efficient, reaches a plateau at 4 weeks in culture, but retains Cx43 expression in contaminating nonastrocytic cells. Our findings indicate that conditional knockout astrocytes are a promising tool for the study of embryonic lethal genes in astrocyte cultures. © 2004 Wiley‐Liss, Inc.
Microglia enhance dorsal root ganglion outgrowth in Schwann cell cultures
Authors: Hynds Dianna L. , Rangappa Nagarathnamma , Beest Julia Ter , Snow Diane M. , Rabchevsky Alexander G.
Published: March 2004

Transplantation of cellular populations to facilitate regrowth of damaged axons is a common experimental therapy for spinal cord injury. Schwann cells (SC) or microglia grafted into injury sites can promote axonal regrowth of central projections of dorsal root ganglion (DRG) sensory neurons. We sought to determine whether the addition of microglia or microglia‐derived secretory products alters DRG axon regrowth upon cultures of SC. Rat DRG explants were grown on monolayers consisting of either SC, microglia, SC exposed to microglia‐conditioned medium (MCM), or co‐cultures with different relative concentrations of microglia. Image analysis revealed that, compared to SC alone, the extent of neurite outgrowth was significantly greater on SC‐microglia co‐cultures. Immunocytochemistry for extracellular matrix molecules showed that microglial cells stained positively for growth‐promoting thrombospondin, whereas laminin and the inhibitory chondroitin sulfate proteoglycans (CSPGs) were localized primarily to SC. Notably, immunoreactivity for CSPGs appeared reduced in areas associated with DRG outgrowth in co‐cultures and SC exposed to MCM. These results show that microglia or their secreted products can augment SC‐mediated DRG regrowth in vitro, indicating that co‐grafting SC with microglia provides a novel approach to augment sensory fiber regeneration after spinal cord injury. © 2004 Wiley‐Liss, Inc.
Translocation of protein kinase C‐βII in astrocytes requires organized actin cytoskeleton and is not accompanied by synchronous RACK1 relocation
Authors: Pascale Alessia , Alkon Daniel L. , Grimaldi Maurizio
Published: March 2004

Protein kinase C (PKC)‐βII is the most abundant PKC isoform in astrocytes. Upon activation, this isoform of PKC translocates from the cytosol to the plasma membrane (PM). In this study, we investigated in astrocytes the modality of PKC‐βII translocation as far as the participation of the receptor for activated C kinase‐1 (RACK1) and the requirement for intact cytoskeleton in the process. In astrocytes, Western blots and immunocytochemistry coupled to confocal microscopic quantitative analysis showed that after 5 min of phorbol‐12‐myristate‐13‐acetate (PMA) exposure, native PKC‐βII, but not PKC‐βI, is relocated efficiently from the cytosol to the PM. Translocation of PKC‐βII was not associated with synchronous RACK1 relocation. Furthermore, the quantity of PM‐associated PKC‐βII that co‐immunoprecipitated with PM‐bound RACK1 increased following PMA exposure, indicating a post activation binding of the two proteins in the PM. Because RACK1 and PKC‐βII relocation seemed not to be synchronous, we hypothesized that an intermediate interaction with the cytoskeleton was taking place. In fact, we were able to show that pharmacological disruption of actin‐based cytoskeleton greatly deranged PKC‐βII translocation to the PM. The requirement for intact actin cytoskeleton was specific, because depolymerization of tubulin had no effect on the ability of the kinase to translocate to the PM. These results indicate that in astrocytes, RACK1 and PKC‐βII synchronous relocation is not essential for relocation of PKC‐βII to the PM. In addition, we show for the first time that the integrity of the actin cytoskeleton plays a specific role in PKC‐βII movements in these cells. We hypothesize that in glial cells, rapidly occurring changes of actin cytoskeleton arrangement may be involved in the fast reprogramming of PKC targeting to specific PM location to phosphorylate substrates in different cellular locations. Published 2003 Wiley‐Liss, Inc.
Apotransferrin promotes the differentiation of two oligodendroglial cell lines
Authors: Paez Pablo M. , García Corina I. , Davio Carlos , Campagnoni Anthony T. , Soto Eduardo F. , Pasquini Juana M.
Published: March 2004

We have previously shown that addition of apotransferrin (aTf) accelerates maturation of oligodendroglial cells (OLGcs) in primary cultures. In this work, we examined the effect of aTf on two conditionally immortalized cell lines: N19 and N20.1. These cells proliferate at 34°C and differentiate into mature OLGcs at 39°C. In vitro addition of aTf to both cell lines at the differentiation temperature for 7 days showed increased expression of galactocerebroside, O4, and myelin basic protein (MBP) and a drop in the percentage of BrdU+ cells. The effect on MBP expression was particularly interesting in the less mature N19 cells. These cells do not express either MBP mRNAs or proteins, so aTf induced, rather than modulated, MBP expression in this cell line. In addition, even though MBP mRNAs for all four isoforms were induced, only the 17 and 21.5 kDa appeared to be translated. OLGc differentiation has been shown to be stimulated by the cAMP‐CREB pathway. In N19 cells, following a pulse of aTf, there was a 10‐fold increase in cAMP levels accompanied by elevated levels of pCREB. In the more mature N20.1 cells, there were no changes in cAMP levels. We conclude that addition of aTf to immature OLGc lines can enhance their expression of differentiated markers, such as MBP. The action of aTf on MBP gene expression in the least mature line is likely to be mediated by the cAMP pathway. In the N20.1 cells, it appears that different signals and/or mechanisms are involved in modulating myelin lipid and MBP expression. The results suggest that aTf can influence OLGc gene expression and differentiation through multiple mechanisms depending on the maturation of the cell. © 2004 Wiley‐Liss, Inc.
GABAA receptors in Müller glial cells of the human retina
Authors: Biedermann Bernd , Bringmann Andreas , Franze Kristian , Faude Frank , Wiedemann Peter , Reichenbach Andreas
Published: March 2004

The present study was aimed at characterizing the GABAA receptor‐mediated currents in acutely isolated glial (Müller) cells of the human retina and investigating their subcellular localization across the Müller cell membrane. Extracellular application of GABA evoked two current responses in human Müller cells: a fast transient GABAA receptor‐mediated current that inactivated within 10 s and that was independent of extracellular Na+, and a sustained current that was dependent on extracellular Na+ and that was mediated by high‐affinity GABA transporters. The receptor current was half‐maximally activated at a GABA concentration of 32 μM, while the transporter current showed an affinity constant of 7.9 μM GABA. The receptor currents were blocked by bicuculline and picrotoxin and were also activated by muscimol or by other amino acids. The receptor currents are Cl− currents, as indicated by the close relationship between the reversal potential of these currents and the Cl− equilibrium potential. Using perforated‐patch recordings, a mean intracellular Cl− concentration of 37 ± 12 mM was determined in human Müller cells. Using electrophysiological and fluorescence imaging methods, it was revealed that GABAA receptors are unevenly distributed across the Müller cell membrane, with higher densities at the endfoot, at the soma, and at the distal sclerad end of the cells. It is concluded that GABAA receptor expression may allow a sensing of retinal GABAergic neuronal signal transmission by Müller cells. © 2004 Wiley‐Liss, Inc.
Astrocyte modulation of in vitro β‐amyloid neurotoxicity
Authors: Paradisi Silvia , Sacchetti Benedetto , Balduzzi Maria , Gaudi Simona , Malchiodi‐Albedi Fiorella
Published: March 2004

In Alzheimer's disease brain, β‐amyloid (Aβ) deposition is accompanied by astrocyte activation, whose role in the pathogenesis of the disease is still unclear. To explore the subject, we compared Aβ neurotoxicity in pure hippocampal cultures and neuronal‐astrocytic cocultures, where astrocytes conditioned neurons but were not in contact with them or Aβ. In the presence of astrocytes, neurons were protected from Aβ neurotoxicity. Neuritic dystrophy was reduced, synapses were partially preserved, and apoptosis was contrasted. The protection disappeared when astrocytes were also treated with Aβ, suggesting that Aβ‐astrocyte interaction is deleterious for neurons. This was supported by comparing Aβ neurotoxicity in pure neurons and neurons grown on astrocytes. In this case, where astrocytes were also in contact with Aβ, neuritic damage was enhanced and expression of synaptic vesicle proteins decreased. Our results suggest that astrocytes can protect neurons from Aβ neurotoxicity, but when they interact with Aβ, the protection is undermined and neurotoxicity enhanced. © 2004 Wiley‐Liss, Inc.
Radial migration of developing microglial cells in quail retina: A confocal microscopy study
Authors: Sánchez‐López Ana , Cuadros Miguel A. , Calvente Ruth , Tassi Mohamed , Marín‐Teva José L. , Navascués Julio
Published: March 2004

Microglial cells spread within the nervous system by tangential and radial migration. The cellular mechanism of tangential migration of microglia has been described in the quail retina but the mechanism of their radial migration has not been studied. In this work, we clarify some aspects of this mechanism by analyzing morphological features of microglial cells at different steps of their radial migration in the quail retina. Microglial cells migrate in the vitreal half of the retina by successive jumps from the vitreal border to progressively more scleral levels located at the vitreal border, intermediate regions, and scleral border of the inner plexiform layer (IPL). The cellular mechanism used for each jump consists of the emission of a leading thin radial process that ramifies at a more scleral level before retraction of the rear of the cell. Hence, radial migration and ramification of microglial cells are simultaneous events. Once at the scleral border of the IPL, microglial cells migrate through the inner nuclear layer to the outer plexiform layer by another mechanism: they retract cell processes, become round, and squeeze through neuronal bodies. Microglial cells use radial processes of s‐laminin‐expressing Müller cells as substratum for radial migration. Levels where microglial cells stop and ramify at each jump are always interfaces between retinal strata with strong tenascin immunostaining and strata showing weak or no tenascin immunoreactivity. When microglial cell radial migration ends, tenascin immunostaining is no longer present in the retina. These findings suggest that tenascin plays a role in the stopping and ramification of radially migrating microglial cells. © 2004 Wiley‐Liss, Inc.
Paranodal axoglial junction is required for the maintenance of the Nav1.6‐type sodium channel in the node of Ranvier in the optic nerves but not in peripheral nerve fibers in the sulfatide‐deficient mice
Authors: Suzuki Ayaka , Hoshi Tomiko , Ishibashi Tomoko , Hayashi Akiko , Yamaguchi Yoshihide , Baba Hiroko
Published: March 2004

In myelinated axons, voltage‐gated sodium channels specifically cluster at the nodes of Ranvier, while voltage‐gated potassium channels are located at the juxtaparanodes. These characteristic localizations are influenced by myelination. During development, Nav1.2 first appears in the predicted nodes during myelination, and Nav1.6 replaces it in the mature nodes. Such replacements may be important physiologically. We examined the influence of the paranodal junction on switching of sodium channel subunits using the sulfatide‐deficient mouse. This mutant displayed disruption of paranodal axoglial junctions and altered nodal lengths and channel distributions. The initial switching of Nav1.2 to Nav1.6 occurred in the mutant optic nerves; however, the number of Nav1.2‐positive clusters was significantly higher than in wild‐type mice. Although no signs of demyelination were observed at least up to 36 weeks of age, sodium channel clusters decreased markedly with age. Interestingly, Nav1.2 stayed in some of the nodal regions, especially where the nodal lengths were elongated, while Nav1.6 tended to remain in the normal‐length nodes. The results in the mutant optic nerves suggested that paranodal junction formation may be necessary for complete replacement of nodal Nav1.2 to Nav1.6 during development as well as maintenance of Nav1.6 clusters at the nodes. Such subtype abnormality was not observed in the sciatic nerve, where paranodal disruption was observed. Thus, the paranodal junction significantly influences the retention of Nav1.6 in the node, which is followed by disorganization of nodal structures. However, its importance may differ between the central and peripheral nervous system. © 2004 Wiley‐Liss, Inc.
Rate‐distortion optimized intra update for error resilience in MPEG‐4 video coding
Authors: Kim Woo‐Shik , Park Rae‐Hong
Published: March 2004

Motion compensation is a powerful method to compress a video sequence. However, once an error occurs, the error propagates through the frames. Recently, the intra update method was proposed to stop the error propagation at the cost of the compression efficiency. This paper proposes an intra update method based on rate‐distortion optimization in error prone environments. The rate and the distortion are estimated using the Lagrangian optimization method to simultaneously select the coding mode and the quantization step size. The proposed method is applied to moving picture experts group (MPEG)‐4 codec, and experimental results show that it is robuster to the packet loss than the conventional ones. © 2004 Wiley Periodicals, Inc. Int J Imaging Syst Technol 13, 341–347, 2003; Published online in Wiley InterScience (www.interscience.wiley.com). DOI 10.1002/ima.10064
Block‐based fine granularity scalable video coding with optimized rate allocation for content‐aware streaming
Authors: He Yuwen , Yang Shiqiang , Zhong Yuzhuo
Published: March 2004

This article proposes a novel block‐based fine granularity scalable (FGS) video coding structure with optimized rate allocation for content‐aware streaming, which is more flexible than MPEG‐4 FGS specified in the streaming profile of MPEG‐4 standard (IS‐14496). In MPEG‐4 FGS streaming, rate allocation can only be based on frame because MBs' compressed data are interlaced between bit‐planes. However in the proposed coding structure, every MB's data are independent; therefore, streaming server can allocate bits according to content's importance, and the quality of those regions of interest is enhanced selectively with more bits allocated. In this way, the streaming server can have a differentiated delivery strategy to support a personalized streaming service considering user's preference. But the uniform rate allocation of proposed block‐based FGS coding will result in a greater loss of coding efficiency than that of MPEG‐4 FGS within a quite wide bit‐rate range. An optimized and fast rate allocation method according to block's rate‐distortion is proposed to solve this critical issue. And the coding efficiency is improved successfully, which can be comparable with MPEG‐4 FGS coding and is even better up to 0.5 dB, with some sequences at medium bit‐rates. © 2004 Wiley Periodicals, Inc. Int J Imaging Syst Technol 13, 322–330, 2003; Published online in Wiley InterScience (www.interscience.wiley.com). DOI 10.1002/ima.10066
Decompositions of complete graphs into triangles and Hamilton cycles
Authors: Bryant Darryn , Maenhaut Barbara
Published: March 2004

For all odd integersn ≥ 1, letGndenote the complete graph of ordern, and for all even integersn ≥ 2 letGndenote the complete graph of ordernwith the edges of a 1‐factor removed. It is shown that for all non‐negative integershandtand all positive integersn,Gncan be decomposed intohHamilton cycles andttriangles if and only ifnh + 3tis the number of edges inGn. © 2004 Wiley Periodicals, Inc.
The nucleocytoplasmic shuttling of E2F4 is involved in the regulation of human intestinal epithelial cell proliferation and differentiation
Authors: Deschênes Claude , Alvarez Laetitia , Lizotte Marie‐Ève , Vézina Anne , Rivard Nathalie
Published: March 2004

The specific mechanisms controlling the transition from proliferation to terminal differentiation in human intestinal epithelial cells (HIEC) remain largely undefined. Herein, we analyzed the expression and localization of Rb and E2F proteins in well‐established normal intestinal epithelial cell models which allow for the re‐enactment of the crypt‐villus axis in vitro as well as in intact epithelium and in colon cancer cells. We report that (1) expression of E2F1 is down‐regulated while E2F4 protein is sequestered in the cytoplasm during G0 arrest associated with serum deprivation, confluency, and terminal differentiation of intestinal cells; (2) concurrently, there is an accumulation of the hypophosphorylated form of the pocket proteins into the nucleus with an increased association of E2F4 with pRb and p130; (3) cells which expressed high levels of nuclear E2F4 are all positive for Ki67 staining in human fetal intestine; (4) activation of HIEC crypt cells by growth factors leads to an increase in the nuclear localization of E2F4 which may be attributable to a decrease in the serine/threonine phosphorylation of this transcription factor; (5) inhibition of p38 MAP kinase with α/β inhibitor SB203580 induces E2F4 translocation into the nucleus and its transcriptional activity. In conclusion, our data suggest a key role for E2F4 in proliferation of human intestinal crypt cells and that its cytoplasmic retention as well as its sequestration by Rb proteins may represent a critical step in initiating cell‐cycle exit. J. Cell. Physiol. 199: 262–273, 2004© 2003 Wiley‐Liss, Inc.
Ultrastructural changes in the interstitial cells of Cajal and gastric dysrhythmias in mice lacking full‐length dystrophin (mdx mice)
Authors: Vannucchi Maria‐Giuliana , Zizzo Maria‐Grazia , Zardo Claudio , Pieri Laura , Serio Rosa , Mulè Flavia , Faussone‐Pellegrini Maria‐Simonetta
Published: March 2004

At least two populations of c‐kit positive interstitial cells of Cajal (ICC) lie in the gastric wall, one located at the myenteric plexus level has a pace‐making function and the other located intramuscularly is intermediary in the neurotransmission and regenerates the slow waves. Both of these ICC sub‐types express full‐length dystrophin. Mdx mice, an animal model lacking in full‐length dystrophin and used to study Duchenne muscular dystrophy (DMD), show gastric dismotilities. The aim of the present study was to verify in mdx mice whether: (i) gastric ICC undergo morphological changes, through immunohistochemical and ultrastructural analyses; and (ii) there are alterations in the electrical activity, using intracellular recording technique. In control mice, ICC sub‐types showed heterogeneous ultrastructural features, either intramuscularly or at the myenteric plexus level. In mdx mice, all of the ICC sub‐types underwent important changes: coated vesicles were significantly more numerous and caveolae significantly fewer than in control; moreover, cytoskeleton and smooth endoplasmic reticulum were reduced and mitochondria enlarged. c‐Kit‐positivity and integrity of the ICC networks were maintained. In the circular muscle of normal mice slow waves, which consisted of initial and secondary components, occurred with a regular frequency. In mdx mice, slow waves occurred in a highly dysrhythmic fashion and they lacked a secondary component. We conclude that the lack of the full‐length dystrophin is associated with ultrastructural modifications of gastric ICC, most of which can be interpreted as signs of new membrane formation and altered Ca2+ handling, and with defective generation and regeneration of slow wave activity. J. Cell. Physiol. 199: 293–309, 2004© 2003 Wiley‐Liss, Inc.
Recovery of cadmium from a zinc hydrometallurgical leachate using reactive emulsion liquid membrane technology
Authors: Fang Jianzhang , Tang Bin , Li Minyu , Xu Zhenghe
Published: March 2004

A new emulsion liquid membrane process using 3,5‐diisopropylsalicylic acid (DIPSA) and triisobutylphosphine sulfide (TIBPS) as carriers, and ammonium sulfide (NH4)2S as precipitant is described. The reactive nature of sulfide ions with extracted cadmium ions in the internal aqueous phase significantly increases cadmium recovery and minimizes zinc impurities. The new process is applied to the enrichment of a low concentration of cadmium ions from a solution containing a high concentration of zinc ions. Under optimum operating conditions, a single stage process produced a cadmium recovery of 98% at a cadmium sulfide content of 99.6%. The results are encouraging for potential applications in zinc hydrometallurgy for recovery of cadmium from sulfuric acid leaching solution of zinc ores. Copyright © 2004 Society of Chemical Industry
Thermal deactivation and inhibition of D‐Amino acid oxidase in permeabilized cells of the yeast Trigonopsis variabilis
Authors: Moreno José A , Ruiz César A , Catalán Jacinto , Galán Miguel A
Published: March 2004

The inhibition of D‐amino acid oxidase contained in permeabilized cells of the yeast Trigonopsis variabilis by α‐keto acids (pyruvic acid, phenylpyruvic acid and 4‐methylthio‐2‐oxobutanoic acid), products of the transformation of the corresponding D‐amino acids, was studied. In all cases, inhibition was of the mixed type and significant differences with respect to the inhibition shown by the enzyme from other sources such as pig kidney or the yeast Rhodotorula gracilis were observed. A study was also made of the thermal deactivation of the enzyme contained in permeabilized cells of T variabilis in the temperature range 30–50 °C in sodium phosphate and Tris hydroxylmethyl aminomethane + CaCl2 buffers. A deactivation mechanism with two steps in series is proposed to account for the variation in activity with time. The results suggest that the enzyme shows greater stability in phosphate buffer, with half‐lives between 7.6 days at 30 °C and 8.6 h at 50 °C. Copyright © 2004 Society of Chemical Industry
Recovery of 2,4‐dichlorophenol from acidic aqueous streams by Membrane Aromatic Recovery System (MARS)
Authors: Dastgir Muhammad G , Ferreira Frederico C , Peeva Ludmila G , Livingston Andrew G
Published: March 2004

This study describes the successful recovery of 2,4‐dichlorophenol (DCP) from wastewater using the Membrane Aromatic Recovery System (MARS). In the MARS process a non‐porous membrane separates a wastewater stream and a stripping solution. DCP is extracted from the wastewater and concentrated in its ionic form in the stripping solution, with pH ≫ pKa DCP. The MARS extraction stage was operated in batch mode with the stripping solution placed inside, and the wastewater stream outside, the membrane tubes. Advantages of this configuration are avoidance of membrane blockage, reduction of stripping solution volume and operational flexibility. The stability and mass‐transfer characteristics of two different membrane materials, poly(dimethylsiloxane) (PDMS) and ethylene–propylene diene terpolymer (EPDM), were tested in DCP solutions with different acidities in order to simulate real industrial waste streams. EPDM exhibits one order of magnitude lower mass‐transfer rates than PDMS (1.4 × 10−7 m s−1 vs 20 × 10−7 m s−1 at 30 °C and 2.4 × 10−7 m s−1 vs 39 × 10−7 m s−1 at 60 °C), however its higher resistance to acid attack provides higher membrane lifetimes. This can be crucial for MARS processes treating real acidic industrial wastewater. A 97% recovery of DCP with a water content of 15 wt% was obtained upon neutralisation of the stripping solution. Copyright © 2004 Society of Chemical Industry
Use of the wings in manipulative and suspensory behaviors during feeding by frugivorous bats
Authors: Vandoros Jason Demetri , Dumont Elizabeth Rachel
Published: March 2004

Frugivory evolved independently in Old and New World fruit bats (Families Pteropodidae and Phyllostomidae, respectively) and anecdotal reports state that these bats use their wings in different ways for manipulating food items and postural support during feeding. However, these often‐cited behavioral differences have not been documented systematically. Here we report observations of manipulative and suspensory behavior collected from 41 individuals representing five phyllostomid and six pteropodid species. During feeding, phyllostomids used both feet to suspend themselves and invariably manipulated food with the wrists and thumbs of both wings. Most pteropodids in our sample used their thumbs for suspension during feeding and none manipulated fruit with their wings. The suspensory and feeding behaviors of pteropodids varied widely and there were significant differences between species. Discrepancies between phyllostomids and pteropodids in the use of the wings during feeding are associated with previously reported differences in wrist morphology. Based on examination of manipulative and suspensory behaviors in a phylogenetic context, we suggest that differences between pteropodids and phyllostomids reflect the distinct ancestral conditions from which these bats evolved. J. Exp. Zool. 301A:361–366, 2004. © 2004 Wiley‐Liss, Inc.
Histology and ultrastructure of male mullerian gland of Uraeotyphlus narayani (Amphibia: Gymnophiona)
Authors: George Jancy M. , Smita Mathew , Oommen Oommen V. , Akbarsha M.A.
Published: March 2004

This study reports the anatomy, histology, and ultrastructure of the male Mullerian gland of the caecilian Uraeotyphlus narayani, based on dissections, light microscopic histological and histochemical preparations, and transmission electron microscopic observations. The posterior end of the Mullerian duct and the urinogenital duct of this caecilian join to form a common duct before opening into the cloaca. The boundary of the entire gland has a pleuroperitoneum, followed by smooth muscle fibers and connective tissue. The Mullerian gland is composed of numerous individual tubular glands separated from each other by connective tissue. Each gland has a duct, which joins the central Mullerian duct. The ducts of the tubular glands are also surrounded by abundant connective tissue. The tubular glands differ between the column and the base in regard to the outer boundary and the epithelial organization. The basement membrane of the column is so thick that amoeboid cells may not penetrate it, whereas that around the base of the gland is thin and appears to allow migration of amoeboid cells into and out of the basal aspect of the gland. The epithelium of the column has nonciliated secretory cells with basal nuclei and ciliated nonsecretory cells with apical nuclei. In the epithelium of the base there are secretory cells, ciliated cells, and amoeboid cells. The epithelium of ducts of the tubular glands is formed of ciliated dark cells and microvillated light cells. The epithelium of the central duct is formed of ciliated dark cells also possessing microvilli, ciliated light cells also possessing microvilli, and microvillated light cells that lack cilia. It is regressed during March to June when the testis lobes are in a state of quiescence. The Mullerian gland is active in secretion during July to February when the testis is active in spermatogenesis. J. Morphol. 260:33–56, 2004. © 2004 Wiley‐Liss, Inc.
Morphology and function of male genitalia (spermatophores) in Euscorpius italicus (Euscorpiidae, Scorpiones): Complex spermatophore structures enable safe sperm transfer
Authors: Jacob Alain , Gantenbein Iris , Braunwalder Matt E. , Nentwig Wolfgang , Kropf Christian
Published: March 2004

The structure and function of the spermatophore of Euscorpius italicus are analyzed. We show how the spermatophore gets shaped from two hemispermatophores and for the first time the sperm transfer mechanism is shown in detail, illustrating function and importance of all complex lobe structures of an euscorpiid spermatophore. A detailed description of the interaction of spermatophore and female genitalia is given. The capsular region of the spermatophore bears different lobes: The distal and basal lobes hook into two cavities on the inner side of the female's genital operculum. A so‐called “crown‐like structure” hooks into a membranous area in the genital atrium. During sperm transfer, these crown‐like structures move backwards, in this way widening the female's genital opening. The sperm duct of the spermatophore is coated with numerous spicules on its outer side, which could serve as a sealing mechanism and/or may stimulate the female. Furthermore, we conclude that “safeguarding of sperm transfer” is one driving force for evolution of male genital complexity in scorpions, but also sexual selection by cryptic female choice could partly play a role. J. Morphol. 260:72–84, 2004. © 2004 Wiley‐Liss, Inc.
Distribution of serotonin in the central nervous system of the blood‐feeding heteropteran, Triatoma infestans (Heteroptera: Reduviidae)
Authors: Settembrini Beatriz P. , Villar Marcelo J.
Published: March 2004

The distribution of serotonin was studied in the Triatoma infestans central nervous system by using immunocytochemistry. Serotonin immunoreactive cell bodies and fibers were observed in the brain, subesophageal ganglion, and thoracic ganglia. In the brain, serotonin‐like immunoreactivity was detected in a limited number of somata, which gave rise to an extensive network of labeled neurites in patterned as well as in nonglomerular neuropils. Immunolabeled perikarya were observed in the optic lobe and in the anteromedial and caudolateral soma rinds of the protocerebrum. Deutocerebral immunoreactive somata were mainly found in the medial layer surrounding the antennal lobe glomeruli, as well as in relationship to the antennal mechanosensory and motor center. The subesophageal ganglion contained serotonin immunoreactive perikarya of variable sizes and moderate to low density of positive fibers. In the prothoracic ganglion, immunoreactive somata were detected near the cephalic connectives as well as in its caudal end. Serotonin immunoreactive somata and fibers were observed in the posterior ganglion of the thorax, with the abdominal neuromeres harboring the highest number of immunolabeled perikarya. These results show that there is a widespread unique serotonergic system in the CNS of Triatoma infestans and suggest that the indolamine could act as a neuromodulator or as a neurohormone. J. Morphol. 260:21–32, 2004. © 2004 Wiley‐Liss, Inc.
Modifications of the falciform process in the eye of beloniformes (Teleostei: Atherinomorpha): Evolution of a curtain‐like septum in the eye
Authors: Reckel Frank , Melzer Roland R.
Published: March 2004

In order to comparatively analyze curtain‐like septa in the eyes of visually orientated “close‐to‐surface‐predators” among atherinomorph teleosts, we examined the eyes of 24 atherinomorph species under a binocular microscope with regard to the falciform process and related structures in the vitreous cavity. Additionally, falciform process samples were analyzed by transmission electron microscopy. All the studied representatives of the Cyprinodontiformes and Atheriniformes, and of one of the beloniform suborder, Adrianichthyioidei, possess a “typical” processus falciformis. In the eyes of the representatives of the other beloniform suborder, Belonoidei, however, pigmented structures that originate in the region of the optic disc and protrude into the vitreous cavity were noted. In the Hemiramphidae (halfbeaks) and Exocoetidae (flying fishes) these pigmented structures have a more cone‐like shape, whereas in the Belonidae (needlefishes) and Scomberesocidae (sauries) horizontally oriented heavily pigmented curtain‐like septa occur that divide the vitreous cavity dorsoventrally. It is suggested that the “typical” processus falciformis represents a plesiomorphic feature within the Atherinomorpha, whereas the pigmented modifications of the falciform process must be seen as a synapomorphic character state of the Belonoidei. The curtain‐like septum of the Belonidae and Scomberesocidae might have evolved from the cone‐like structures that are found in the Exocoetoidea. The functional significance of the pigmented structures in the eye is as yet not clear, except for the curtain‐like septum found in Belonidae. It might play a role in visual orientation near the water surface at Snell's window. J. Morphol. 260:13–20, 2004. © 2004 Wiley‐Liss, Inc.
Regional outbreak of mumps due to genotype H in Korea in 1999
Authors: Lee Joo‐Yeon , Na Byoung‐Kuk , Kim Jee‐Hee , Lee Jin‐Soo , Park Jong‐Won , Shin Gu‐Choul , Cho Hae‐Wol , Lee Ho‐Dong , Gou Un‐Yeong , Yang Byoung‐Kuk , Kim Joon , Kang Chun , Kim Woo‐Joo
Published: March 2004

Serological and virological studies were carried out of a mumps outbreak which occurred in one region, Yoeju County, Southeast of Seoul in Korea from September to December, 1999. Sera from 736 children at 8–13 years of age of patients with mumps and healthy children were tested for mumps‐specific antibodies by enzyme immunoassay. The overall IgM positive rate was 7.6% (56/736), compared with 69.8% (514/736) for IgG. Of the 49 children with both IgG and IgM, 32 were also confirmed by both clinical and serological diagnosis. IgM antibodies were detected even in the samples collected up to 3 months after the onset of symptoms. Although 436 children had been vaccinated before the outbreak, 27 (6.2%) were found to be IgM positive, particularly 6 (4.4%) of 136 were positive serologically despite a second‐dose vaccinees. Sequence analysis of the small hydrophobic (SH) gene of 4 mumps viruses isolated from 42 saliva specimens revealed that these were related to the genotype H, but distinguishable from European strains. This is the first study on the outbreak due to mumps virus genotype H and provides information to assess the understanding of recent outbreaks of mumps in Korea. J. Med. Virol. 73:85–90, 2004. © 2004 Wiley‐Liss, Inc.
Genetic variation in clinical varicella‐zoster virus isolates collected in Ireland between 2002 and 2003
Authors: Carr Michael J. , McCormack Grace P. , Crowley Brendan
Published: March 2004

Analysis of genetic variation in 16 varicella‐zoster virus (VZV) isolates selected at random and circulating in the Irish population between March 2002 and February 2003 was carried out. A 919 bp fragment of the glycoprotein E gene (open reading frame 68) encompassing codon 150, at which a non‐synonymous mutation defines the escape mutant VZV‐MSP, and including two other epitope regions e1 and c1, was sequenced. No new single nucleotide polymorphisms (SNPs) were detected, indicating stability of these epitopes in clinical isolates of VZV. However, when four informative polymorphic markers consisting of defined regions from genes 1, 21, 50, and 54 were sequenced 14 variable nucleotide positions were identified. Phylogenetic analysis showed the presence of three highly supported clades A, B, and C circulating in the Irish population. Approximately one third (6/16; 37.5%) of the Irish VZV isolates in this study belonged to genotype C, 4/16 (25%) to genotype A, and 4/16 (25%) to genotype B. A smaller number 2/16 (12.5%) belonged to genotype J1. This indicates remarkable heterogeneity in the Irish population given the small sample size. No evidence was found to suggest any of the 16 isolates was a recombinant. These findings have implications for the model of geographic isolation of VZV clades to certain regions as the circulating Irish VZV population appears to comprise approximately equal numbers of each of the main genotypes. This data is inconsistent with a model of strict geographical separation of VZV genotypes and suggests that VZV diversity is more pronounced in certain areas than had been thought previously. J. Med. Virol. 73:131–136, 2004. © 2004 Wiley‐Liss, Inc.
Comparative evaluation of measles virus specific TaqMan PCR and conventional PCR using synthetic and natural RNA templates
Authors: Ozoemena L.C. , Minor P.D. , Afzal M.A.
Published: March 2004

Comparative evaluation of TaqMan RT‐polymerase chain reaction (PCR) methodology developed during this study with the conventional RT‐PCR‐nested PCR methodology developed earlier, using measles virus RNA templates derived from synthetic and natural sources against a number of primer sets belonging to various regions of the genome, revealed the existence of similar assay thresholds for both methods. An exception to this finding was, however, noted using primer sets of the N and M genes regions with RNA templates extracted from the wild type measles virus strain where the nested PCR method proved to be 10‐ to 100‐fold more sensitive than the end points established with the N gene specific TaqMan RT‐PCR method with synthetic RNA templates. These differences were not evident when the same primer sets were evaluated with RNA templates extracted from a brain sample of SSPE patient. These findings indicate that the genetic make up of measles virus strain in any given clinical specimen, in relation to the amplifying primers/probe sequences, can have impact on the overall sensitivity and specificity of the methodology applied. Both methods are equally suitable for the molecular detection of measles virus sequences in clinical specimens, although the TaqMan RT‐PCR method may be preferred due to its advantages of contamination control, automation, and real‐time product quantitation. J. Med. Virol. 73:79–84, 2004. © 2004 Wiley‐Liss, Inc.
Trends in HIV prevalence among drug‐users attending public drug‐treatment centres in Italy: 1990–2000
Authors: Suligoi Barbara , Magliochetti Natalia , Nicoletti Giovanni , Pezzotti Patrizio , Rezza Giovanni
Published: March 2004

The occurrence of HIV infection among drug‐users in Italy between 1990 and 2000 is described; the data concern drug‐users attending a nation‐wide network consisting of 510 public drug‐treatment centres. We included only individuals with written documentation of an HIV‐test result. Of the 1,299,972 attendees, 54.5% had a documented HIV‐test result. The overall HIV prevalence among those with a test result was 19.8%. The annual prevalence decreased from 30.8% in 1990 to 15.8% in 2000 (chi‐square for linear trend P < 0.0001); the decrease was more evident between 1990 and 1996, whereas the prevalence apparently levelled‐off after 1996. The annual prevalence was significantly higher among females compared to males throughout the study period. The annual prevalence was consistently three to four times higher for prior attendees, compared to new entrants. Although information on whether or not the attendees injected drugs was not available for the entire study period, the proportion of attendees with a documented HIV‐test result can be considered as an approximation of the proportion who inject drugs. Although HIV infection was widespread among injecting drug‐users in Italy beginning in the mid‐1980s, since the early 1990s it has been somewhat contained. However, the levelling‐off of the prevalence after 1996 may conceal an increase in the number of new infections and should be more thoroughly investigated. Aggregated national‐level data, despite their limitations, represent a reliable tool for monitoring the temporal trend of HIV prevalence among injecting drug‐users in Italy. J. Med. Virol. 73:1–6, 2004. © 2004 Wiley‐Liss, Inc.
Anti‐envelope 1 and 2 immune response in chronic hepatitis C patients: Effects of hepatitis B virus co‐infection and interferon treatment
Authors: Zampino Rosa , Marrone Aldo , Mangoni Emanuele Durante , Santarpia Lucio , Sica Antonello , Tripodi Marie‐Francoise , Utili Riccardo , Ruggiero Giuseppe , Adinolfi Luigi Elio
Published: March 2004

Antibodies against envelope glycoprotein 1 and 2 (anti‐E1/E2) have been suggested to influence HCV replication levels. Hepatitis B virus (HBV) may interfere with hepatitis C virus (HCV) replication. At present there are no data on anti‐E1/E2 antibody responses or on the effect of interferon (IFN) treatment in HBV–HCV co‐infection. Accordingly, we evaluated serum anti‐E1/E2 antibodies in 50 patients (median age, 26.5; males, 30) with chronic hepatitis, 38 with HCV and 12 with HBV–HCV co‐infection, who had undergone α‐IFN treatment. Before starting IFN, the HCV group showed higher HCV‐RNA levels (bDNA assay) than the HBV–HCV group (median 3.75 vs. 0.64 × 106 Eq/ml, respectively; P < 0.05). Similarly, the anti‐E2 levels (EIA assay) were higher in the HCV group than in the HBV–HCV (mean ± SD, 53.8 ± 54.58 vs. 24.5 ± 41.50 U/ml, respectively; P < 0.02), and the prevalence of anti‐E2 was also higher in the HCV group (94 vs. 58%, respectively; P < 0.007). No correlation was found between anti‐E1/E2 antibodies and the HCV‐RNA levels. The prevalence of E1/E2 antibodies was similar in the different HCV genotypes. Higher baseline levels of anti‐E2 antibodies and a decrease or disappearance of anti‐E2 antibodies during IFN were associated with IFN sustained response in both groups, whereas no reduction in the anti‐E1/E2 levels was observed in non‐responders. The data show that HBV co‐infection influences both HCV replication and the anti‐E1/E2 antibody production. High pre‐treatment levels of anti‐E2 antibodies and their decrease or disappearance during interferon treatment are often associated with HCV clearance in sustained responders, irrespective of the HCV genotype. J. Med. Virol. 73:33–37, 2004. © 2004 Wiley‐Liss, Inc.
Characterisation of penciclovir resistant acyclovir sensitive herpes simplex virus type 2 isolated from an AIDS patient
Authors: Pardeiro Mercedes , Cuenca‐Estrella Manuel , Fernández‐Clúa Miguel A. , Santos‐O'Connor Francisco , Tabarés Enrique , Gadea Ignacio
Published: March 2004

A heterogeneous herpes simplex virus type 2 (HSV‐2) population was characterised from an AIDS patient with relapsing genital ulcer. The isolate had an unusual antiviral spectrum, showing resistance to penciclovir and susceptibility to acyclovir. Two viral populations were plaque purified, one resistant and the other susceptible to both antiviral drugs. The resistant clone was deficient in thymidine kinase (TK) activity and a nucleotide substitution, thymine for cytosine, at position 153 was identified in its TK gene. This mutation resulted in an amino acid change, arginine to tryptophan, in the ATP binding site. In the deficient mutant, a loss of virulence was observed in mice. J. Med. Virol. 73:60–64, 2004. © 2004 Wiley‐Liss, Inc.
High rate and changing molecular epidemiology pattern of norovirus infections in sporadic cases and outbreaks of gastroenteritis in Hong Kong
Authors: Lau Chi‐Shan , Wong Derek A. , Tong Louis K.L. , Lo Janice Y.C. , Ma Agnes M.C. , Cheng Peter K.C. , Lim Wilina W.L.
Published: March 2004

Noroviruses (Norwalk‐like viruses (NLV)) are recognised as major causes of acute gastroenteritis worldwide. Numerous studies had been carried out on the molecular epidemiology of norovirus in outbreaks but relatively few on sporadic cases. In this study, the molecular epidemiology of noroviruses in sporadic and outbreak cases of acute gastroenteritis in Hong Kong was examined over a 12‐month period from July 2001 to June 2002. Specimens from three groups of patients were used in this study. Nine hundred ninety‐five specimens from patients enrolled in the Acute Diarrhoeal Diseases Surveillance Programme of the Department of Health, Hong Kong Government; 735 clinical specimens from hospital patients with acute gastroenteritis, and 122 specimens from 44 norovirus outbreaks. Ninety‐two (9.2%) surveillance specimens were positive for norovirus RNA by reverse transcription‐polymerase chain reaction (RT‐PCR), compared to 123 (16.7%) clinical and 101 (82.8%) outbreak specimens. For the first 6 months of the study period, the predominant strain was the Bristol strain that belongs to genogroup II (GII). In the latter 6 months of the study, genogroup I (GI) and strains belonging to other clusters of GII were seen more commonly. The vast majority of strains belonging to the Bristol virus cluster were closely related to the 95/96‐US subset that was associated with pandemic infection from 1995 onwards. This study clearly establishes the importance of norovirus as a cause of sporadic cases of acute gastroenteritis in all age groups in Hong Kong. J. Med. Virol. 73:113–117, 2004. © 2004 Wiley‐Liss, Inc.
PCR‐probe capture hybridization assay and statistical model for SEN virus prevalence estimation
Authors: Tanaka Yasuhito , Pfeiffer Ruth , Yeo Anthony E.T. , Mizokami Masashi , Edlin Brian R. , Castle Philip , O'Brien Thomas R. , Alter Harvey J. , Shih J. Wai‐Kuo
Published: March 2004

SEN viruses (SENV) are newly discovered blood‐borne single‐stranded circular DNA viruses that may play a role in liver disease. To date, no serologic assays are available for the detection of SENV antigens or antibodies. We report on a rapid and sensitive molecular assay for the detection of four SENV strains (SENV‐A, ‐C, ‐D, ‐H). This method uses PCR with universal primers and microwell capture hybridization with type‐specific probes. Cut‐off points to define “infected” based on chemiluminescence readings were determined from a statistical mixture model applied to samples from 300 injection drug users (IDUs) in San Francisco. Based on the estimated cut‐off points, we examined the prevalence of SENV infection among 232 healthy US blood donors and assessed sensitivity and specificity of the assay in a small validation sample of infected individuals with partial sequence information. J. Med. Virol. 73:123–130, 2004. © 2004 Wiley‐Liss, Inc.
Insulin‐like growth factor 1 prevents neuronal cell death induced by corticosterone through activation of the PI3k/Akt pathway
Authors: Nitta Atsumi , Zheng Wen‐Hua , Quirion Rémi
Published: March 2004

Corticosterone (CORT) is well known to induce neuronal damage in various brain regions including the hippocampus, but the precise mechanism(s) of action underlying these effects has yet to be fully established. Insulin‐like growth factor‐1 (IGF‐1) is a trophic factor promoting cell survival by the activation of the phosphatidylinositide 3‐kinase (PI3K)/Akt kinase pathway. We report that IGF‐1 prevents neuronal cell death induced by CORT, likely via the stimulation of the PI3K/Akt pathway in primary hippocampal cultured neurons. CORT induced neuronal cell death at a minimal concentration of 50 nM. IGF‐1 (10 nM) prevented cell death induced by CORT under serum‐free conditions. The neuroprotective effect of IGF‐1 was accompanied by reversal of the Akt pathway inhibition induced by CORT. The PI3 kinase inhibitor, LY29004, inhibited the neuroprotective effect of IGF‐1 whereas the MEK (MAPK kinase) inhibitor PD98059, an upstream blocker of mitogen‐activated protein (MAP) kinase, had no effect. These results suggest that IGF‐1 can prevent neuronal cell death induced by CORT in hippocampal neurons by modulating the activity of the PI3K/Akt pathway. © 2004 Wiley‐Liss, Inc.