bluuebunny/crossref_metadata_2025 huggingface dataset:
Autonomous differentiation of Drosophila spermatogonia in vitro
Authors: Kawamoto Tatsuya , Kawai Keita , Kodama Tatsuji , Yokokura Tomohiro , Niki Yuzo
Published: August 2010

Spermatogenesis is a complex process that produces functional sperm by establishing male germline stem cells (mGSCs) in adult testes. To study Drosophila spermatogenesis in vitro, we examined various culture conditions of spermatogonia. Spermatogonia from larval testes began to differentiate soon after culture, whereas mGSCs did not undergo self‐renewal division. Strikingly, 16‐cell spermatogonia from early and late larval testes differentiated into motile spermatids autonomously. Furthermore, individual spermatogonia developed into motile spermatids even after mechanical dissociation from encapsulating cyst cells. This is the first study to report that spermatogonia in larval testes retain the ability to differentiate into spermatids in the absence of gonadal tissue. Our in vitro system should provide an excellent opportunity to study spermatogenesis in detail and apply genetic manipulation.
Mutation in the abcb7 gene causes abnormal iron and fatty acid metabolism in developing medaka fish
Authors: Miyake Akimitsu , Higashijima Shin‐ichi , Kobayashi Daisuke , Narita Takanori , Jindo Tomoko , Setiamarga Davin H. E. , Ohisa Satoshi , Orihara Nobuaki , Hibiya Kenta , Konno Sayaka , Sakaguchi Sae , Horie Kyohei , Imai Yoshiyuki , Naruse Kiyoshi , Kudo Akira , Takeda Hiroyuki
Published: August 2010

The medaka fish (Oryzias latipes) is an emerging model organism for which a variety of unique developmental mutants have now been generated. Our recent mutagenesis screening of the medaka isolated a unique mutant that develops a fatty liver at larval stages. Positional cloning identified the responsible gene as medaka abcb7. Abcb7, a mitochondrial ABC (ATP binding cassette) half‐transporter, has been implicated in iron metabolism. Recently, human Abcb7 was found to be mutated in X‐linked sideroblastic anemia with cerebellar ataxia (XLSA/A). The homozygous medaka mutant exhibits abnormal iron metabolism in erythrocytes and accumulation of lipid in the liver. Microarray and in situ hybridization analyses demonstrated that the expression of genes involved in iron and lipid metabolisms are both affected in the mutant liver, suggesting novel roles of Abcb7 in the development of physiologically functional liver. The medaka abcb7 mutant thus could provide insights into the pathogenesis of XLSA/A as well as the normal function of the gene.
Regulation of dermal fibroblast dedifferentiation and redifferentiation during wound healing and limb regeneration in the Axolotl
Authors: Satoh Akira , Bryant Susan V. , Gardiner David M.
Published: August 2010

Adult urodeles (salamanders) are unique in their ability to regenerate complex organs perfectly. The Accessory Limb Model (ALM) in the axolotl allows for the identification of signals from the wound epidermis, nerves and dermal fibroblasts that interact to regenerate a limb. In the present study, we have used the ALM to identity the axolotl (Ambystoma mexicanum) orthologue of Twist (AmTwist), a basic helix‐loop‐helix transcription factor that is involved in the regeneration of the dermis during limb regeneration. AmTwist is expressed during the blastema stages in regeneration, but is inhibited by signals from the nerve during the early stages when dermal fibroblasts dedifferentiate to form blastema cells. As the dermis regenerates, AmTwist is expressed in association with the synthesis of type I collagen in the proximal region of the blastema. Exogenous bone morphogenetic protein‐2 leads to an increase in AmTwist expression, and therefore may function as an endogenous regulator of AmTwist expression and dermis regeneration. The nerve appears to have a dual function in regeneration by coordinately regulating dedifferentiation and redifferentiation of dermal fibroblasts.
Reelin, radial fibers and cortical evolution: Insights from comparative analysis of the mammalian and avian telencephalon
Authors: Nomura Tadashi , Hattori Mitsuharu , Osumi Noriko
Published: August 2010

The mammalian cerebral cortex has a remarkable laminated structure, which is derived from the pallium, the dorsal part of the embryonic telencephalon. Recent studies indicate that the pallium is developed as a homologous structure in all vertebrate species. However, the cellular and molecular mechanism for making architectural diversity of the pallium is not fully understood. Here we introduce recent progress in comparative analysis of pallial development, and our data on the role of Reelin protein in the developing avian pallium. These experimental approaches to pallial development in non‐mammalian species will provide a new insight into evolution of the cerebral cortex.
Weaving the neuronal net with target‐derived fibroblast growth factors
Authors: Umemori Hisashi
Published: August 2010

Fibroblast growth factors (FGFs) are a large family of secreted growth factors that are involved in the development, regeneration and repair of various tissues. In the nervous system, FGFs have been implicated in early developmental processes, such as neural induction, proliferation and patterning. Accumulating data indicate that FGFs are also important for the formation of functional neural networks. The role of FGFs in axon guidance, target recognition and synaptic differentiation as target‐derived factors, and how they cooperate with cell adhesion molecules that are also involved in the wiring of the nervous system are the focus of this review.
Development of the larval muscle system in the mussel Mytilus trossulus (Mollusca, Bivalvia)
Authors: Dyachuk Vyacheslav , Odintsova Nelly
Published: August 2010

In the present study we examined muscle development throughout the complete larval cycle of the bivalve mollusc, Mytilus trossulus. An immunofluorescence technique and laser scanning confocal microscopy were used in order to study the organization of the muscle proteins (myosin, paramyosin, twitchin, and actin) and some neurotransmitters. The appearance of the muscle bundles lagged behind their nervous supply: the neuronal elements developed slightly earlier (by 2 h) than the muscle cells. The pioneer muscle cells forming a prototroch muscle ring were observed in a completed trochophore. We documented a well‐organized muscle system that consisted of the muscle ring transforming into three pairs of velar striated retractors in the early veliger. The striations were positive for all muscle proteins tested. Distribution of FMRFamide and serotonin (5‐HT) immunocytochemical staining relative to the muscle ring differed significantly: 5‐HT‐immunioreactive cells were situated in the center of the striated muscle ring, while Phe‐Met‐Arg‐Phe‐NH2 neuropeptide FMRFamid immunoreactive fibers were located in a distal part of this ring. Our data showed clearly that the muscle proteins and the neurotransmitters were co‐expressed in a coordinated fashion in a continuum during the early stages of the mussel development. Our study provides the first strong evidence that mussel larval metamorphosis is accompanied by a massive reorganization of striated muscles, followed by the development of smooth muscles capable of catch‐contraction.
Expression of the congenital heart disease 5/tryptophan rich basic protein homologue gene during heart development in Medaka fish, Oryzias latipes
Authors: Murata Kenji , Degmetich Sean , Kinoshita Masato , Shimada Eriko
Published: August 2010

The congenital heart disease 5 (CHD5)/tryptophan rich basic protein (WRB) is a protein containing a tryptophan‐rich carboxy‐terminal region, which was discovered in the human fetal heart. In humans, this CHD5/WRB is located between the markers ACTL5‐D21S268 within the Down syndrome (DS) Region‐2 at chromosome 21. Congenital heart disease is commonly linked to DS patients. The functions of this gene product are unknown. To identify the functions of CHD5/WRB in heart formation during embryogenesis, the medaka CHD5 cDNA (mCHD5) was isolated and its gene expression pattern and the localization of its gene product were investigated. The obtained mCHD5 belongs to the CHD5 superfamily, whose members include coiled‐coil proteins. The mCHD5 gene was found to be expressed in the developing heart after stage 28 at which the chamber (ventricle and atrium) differentiation in the heart tube is initiated in the embryo. Its gene product was also detected in the developing heart at embryonic stage 28 and 35. Knocking‐down of mCHD5 function caused severe cardiac disorder, including abnormal chamber differentiation, abnormal looping and ocular abnormality such as Cyclops. Our results provide the mCHD5 gene expression pattern as well as its physiological role during heart formation in a vertebrate model system.
Comparative molecular neuroanatomy of mammalian neocortex: What can gene expression tell us about areas and layers?
Authors: Watakabe Akiya
Published: August 2010

It is over 100 years since Brodmann proposed the homology of layer and area structure of the cerebral cortex across species. His proposal was based on the extensive comparative analyses of various mammalian brains. Although such homology is now well accepted, the recent data in our laboratory showed striking variations of gene expression patterns across areas and species. Are cortical layers and areas really homologous? If they are, to what extent and how are they similar or different? We are trying to answer these questions by identifying the homologous neuronal types common to various areas and species. Toward this goal, we started to classify the cortical pyramidal neurons by expression of particular sets of genes. By using fluorescent double in situ hybridization combined with retrograde tracers, we are characterizing the gene expression phenotypes and projection specificity of cortical excitatory neuron types. In this review, I discuss the recent findings in our laboratory in light of the past and present knowledge about cortical cell types, which provides insight to the homology (and lack thereof) of the mammalian neocortical organization.
Development of the central and peripheral nervous systems in the lamprey
Authors: Murakami Yasunori , Watanabe Aki
Published: August 2010

Vertebrate brains are highly organized structures that show remarkable diversity throughout the animal groups. Among the vertebrates, the agnathan animals, which diverged from the gnathostomes early in the evolution of the vertebrates, occupy a key phylogenetic position in order to clarify the origin and evolution of the brain. We found that the lamprey brain has the basic molecular mechanisms necessary to form brain compartments. Conversely, the telencephalon and cerebellum display gnathostome‐specific developmental mechanisms. We also propose that, in contrast to those of gnathostomes, the maxillary ramus of the trigeminal nerve and buccal ramus of the nerve on the anterior lateral line are not fused in the developing lamprey. Thus, the development of the central nervous system and the framework of the peripheral nerve around the oral region are thought to have improved in the course of the agnathan–gnathostome transition.
From brain formation to plasticity: Insights on Otx2 homeoprotein
Authors: Sugiyama Sayaka , Prochiantz Alain , Hensch Takao K.
Published: August 2010

The shaping of neuronal circuits is essential during postnatal brain development. A window of neuronal remodeling by sensory experience typically occurs during a unique time in early life. The many types of behavior and perception, like human language, birdsong, hearing and vision are refined by experience during these distinct ‘critical periods’. The onset of critical periods for vision is delayed in animals that remain in complete darkness from birth. It is then predicted that a ‘messenger’ within the visual pathway signals the amount of sensory experience that has occurred. Our recent results indicate that Otx2 homeoprotein, an essential morphogen for embryonic head formation, is reused later in life as this ‘messenger’ for critical period plasticity. The homeoprotein is stimulated by visual experience to propagate into the visual cortex, where it is internalized by GABAergic interneurons, especially Parvalbumin‐positive cells (PV‐cells). Otx2 promotes the maturation of PV‐cells, consequently activating critical period onset in the visual cortex. Here, we discuss recent data that are beginning to illuminate the physiological function of non‐cell autonomous homeoproteins, as well as the restriction of their transfer to PV‐cells in vivo.
Cerebral cortex development: From progenitors patterning to neocortical size during evolution
Authors: Pierani Alessandra , Wassef Marion
Published: August 2010

The central nervous system is composed of thousands of distinct neurons that are assembled in a highly organized structure. In order to form functional neuronal networks, distinct classes of cells have to be generated in a precise number, in a spatial and temporal hierarchy and to be positioned at specific coordinates. An exquisite coordination of appropriate growth of competent territories and their patterning is required for regionalization and neurogenesis along both the anterior‐posterior and dorso‐ventral axis of the developing nervous system. The neocortex represents the brain territory that has undergone a major increase in its relative size during the course of mammalian evolution. In this review we will discuss how the fine tuning of growth and cell fate patterning plays a crucial role in the achievement of the final size of central nervous system structures and how divergence might have contributed to the surface increase of the cerebral cortex in mammals. In particular, we will describe how lack of precision might have been instrumental to neocortical evolution.
The Sperm‐surface glycoprotein, SGP, is necessary for fertilization in the frog, Xenopus laevis
Authors: Nagai Keita , Ishida Takuya , Hashimoto Takafumi , Harada Yuichirou , Ueno Shuichi , Ueda Yasushi , Kubo Hideo , Iwao Yasuhiro
Published: August 2010

To identify a molecule involved in sperm‐egg plasma membrane binding at fertilization, a monoclonal antibody against a sperm‐surface glycoprotein (SGP) was obtained by immunizing mice with a sperm membrane fraction of the frog, Xenopus laevis, followed by screening of the culture supernatants based on their inhibitory activity against fertilization. The fertilization of both jellied and denuded eggs was effectively inhibited by pretreatment of sperm with intact anti‐SGP antibody as well as its Fab fragment, indicating that the antibody recognizes a molecule on the sperm's surface that is necessary for fertilization. On Western blots, the anti‐SGP antibody recognized large molecules, with molecular masses of 65–150 kDa and minor smaller molecules with masses of 20–28 kDa in the sperm membrane vesicles. SGP was distributed over nearly the entire surface of the sperm, probably as an integral membrane protein in close association with microfilaments. More membrane vesicles containing SGP bound to the surface were found in the animal hemisphere compared with the vegetal hemisphere in unfertilized eggs, but the vesicle‐binding was not observed in fertilized eggs. These results indicate that SGP mediates sperm‐egg membrane binding and is responsible for the establishment of fertilization in Xenopus.
Extra‐mitochondrial localization and likely reproductive function of a female‐transmitted cytochrome c oxidase subunit II protein
Authors: Chakrabarti Rumela , Shepardson Sally , Karmakar Moumita , Trdan Richard , Walker Jennifer , Shandilya Ramdas , Stewart Don , Vijayaraghavan Srinivasan , Hoeh Walter
Published: August 2010

Our previous study documented a reproductive function for the male‐transmitted mitochondrial DNA (mtDNA)‐encoded cytochrome c oxidase subunit II (MCOX2) protein in a unionoid bivalve. Here, immunoblotting, immunohistochemistry and immunoelectron microscopy analyses demonstrate that the female‐transmitted protein (FCOX2) is: (i) expressed in both male and female gonads; (ii) maximally expressed in ovaries just prior to the time of the annual fertilization event; (iii) displayed in the cytoplasm and more strongly in the plasma membrane (microvilli), vitelline matrix and vitelline envelope of mature ovarian eggs; and (iv) strongly localized to the vitelline matrix of some eggs just prior to fertilization. These findings represent evidence for the extra‐mitochondrial localization of an mtDNA‐encoded gene product and are consistent with multifunctionality for FCOX2 in eggs.
The feasibility of non‐viral gene transfer to the diaphragm in vivo
Authors: Beshay Morris , Gazdhar Amiq , Gugger Mathias , Reymond Marc , Schmid Ralph A.
Published: August 2010

Gene transfer using electroporation is an essential method for the study of developmental biology, especially to understand the internal control of degeneration and apoptosis of the muscle cells that occurs earlier and quicker than the usual degeneration process occurring by aging. Such experimental studies may have a role in developing new strategies for treating patients suffering from inherited primary myopathies such as Duchenne muscular dystrophy (DMD). The present study was designed to evaluate the feasibility of electroporation mediated transfer of reporter genes to the diaphragm in vivo. This is the first report of gene transfer of naked plasmid DNA into the diaphragm muscle in vivo using electroporation. Our results showed that in vivo gene transfer of naked plasmid DNA into the diaphragm muscle using electroporation is feasible.
Heterogeneity of mouse primordial germ cells reflecting the distinct status of their differentiation, proliferation and apoptosis can be classified by the expression of cell surface proteins integrin α6 and c‐Kit
Authors: Morita‐Fujimura Yuiko , Tokitake Yuko , Matsui Yasuhisa
Published: August 2010

Primordial germ cells (PGCs) in mouse embryos likely include heterogeneous cells having distinct cellular properties. In the present study, we found that heterogeneity of PGCs can be defined by the expression of integrin α6 and c‐Kit. The changes in integrin α6 and c‐Kit expression in PGCs were obvious as embryonic development progressed, and the PGCs became a mixture of populations consisting of cells with distinct levels of cell surface protein expression. The changes and heterogeneity of cell surface protein expression mainly reflected asynchronous differentiation of PGCs. Apoptosis of PGCs was biased in populations of c‐Kit or integrin α6 negative PGCs at particular developmental stages, suggesting possible linkage between PGC apoptosis and the levels of expression of these cell surface proteins. Histochemical analysis confirmed the heterogeneous expression of c‐Kit and integrin α6 in PGCs in embryonic gonads, and revealed that PGCs showing different levels of integrin α6 or c‐Kit expression and the apoptotic PGCs were scattered and did not show specific localization within gonads. The present study enables us to analyze and isolate populations of living PGCs showing a distinct status of differentiation, or different properties of proliferation or of cell death in individual embryos, and provides a new strategy to examine the mechanisms of PGC development.
Spatio‐temporal expression of activating transcription factor 5 in the skeletal development of mouse limb
Authors: Satake Hiroshi , Ito Kazuo , Takahara Masatoshi , Furukawa Takashi , Takagi Michiaki , Ogino Toshihiko , Shinomura Tamayuki
Published: August 2010

There are no data about the expression pattern of activating transcription factor 5 (Atf5) during limb development. The aim of the present study was to describe the expression of Atf5 throughout mouse limb development. Some specimens were examined for the expression of type II collagen α1 (Col2a1), sex‐determining region Y‐related high‐mobility‐group‐box 9 (Sox9), and activating transcription factor 2 (Atf2) by in situ hybridization. ATDC5 cells, mouse limb buds, and epiphyseal cartilage were examined for the expression of Atf5α and Atf5β by quantitative real‐time PCR. Atf5 transcripts were detected in condensed mesenchymal cells, cartilage primordia, proliferative, prehypertrophic, and articular chondrocytes, but not in hypertrophic chondrocytes. Atf5 was predominantly expressed by osteoblasts in the primary spongiosa. At the postnatal stage, Atf5 was expressed in epiphyseal chondrocytes and osteoblasts lining the bone trabeculae. Spatial distribution of Col2a1, Sox9, and Atf2 transcripts closely corresponded to that of Atf5 within developing limbs, except for bony tissues. In cartilage, Atf5α is the dominant form in the embryonic stage as well as in the postnatal stage.
Duplicated zebrafish relaxin‐3 gene shows a different expression pattern from that of the co‐orthologue gene
Authors: Donizetti Aldo , Fiengo Marcella , Minucci Sergio , Aniello Francesco
Published: August 2010

Relaxin‐3 (Rln3) is thought to function as a neurotransmitter mainly produced in the mammalian nucleus incertus and is involved in different neural processes; among them, the stress response and food intake. Here, we report the expression pattern of the duplicated zebrafish rln3b gene and compare it to the previously analyszd spatial expression pattern of the rln3a gene. Both genes, during the embryogenesis and in the adult fish, are active and show relevant differences in their expression patterns. rln3b is diffusely expressed in the brain until the pharyngula period, when, at 48 h postfertilization (hpf), the expression becomes restricted to the periaqueductal gray, where it persists also at later developmental stages. No expression was observed in the nucleus incertus cells that express the rln3a gene from 72 hpf. In the adult, both genes are expressed in brain, but only rln3b transcript is revealed in testis at the similar expression level, whereas in the other analyzed tissues the transcript levels are lower or absent. Both the putative mature protein sequences are highly conserved, this feature and their differential expression patterns might indicate a sub‐functionalization during evolution with the consequent retention of the two paralogues genes.
Mechanism of DNA replication‐dependent transcriptional activation of the acetylcholinesterase gene in the Ciona intestinalis embryo
Authors: Kataoka Yumiko , Mishina Ryo , Fujiwara Shigeki
Published: August 2010

The acetylcholinesterase‐encoding gene in the ascidian Ciona intestinalis (Ci‐AChE) is expressed in tail muscle cells from the gastrula stage. When the embryo was continuously treated with aphidicolin from the 32‐cell stage, Ci‐AChE was not expressed even when control embryos reached the tailbud stage. This result suggests that Ci‐AChE acquires the competence to be transcribed after passing through a certain number of DNA replication cycles. A lacZ reporter gene containing the 5′ flanking region of Ci‐AChE was expressed in the tail muscle cells. Aphidicolin treatment from the 32‐cell stage affected, but did not completely suppress, the expression of lacZ. A bisulfite sequencing analysis was carried out to examine the methylation status of four regions within the 5′ flanking sequence and the first exon. However, all of these regions remained unmethylated from the 16‐cell to 110‐cell stages. The results suggested that the DNA of the Ci‐AChE locus is not responsible for counting the rounds of replication. We examined the expression of the C. intestinalis MyoD (Ci‐MyoD), a transcription factor that activates Ci‐AChE. Aphidicolin treatment from the 32‐cell stage suppressed the expression of Ci‐MyoD, even when control embryos reached the gastrula stage. These results suggest that a lack of Ci‐MyoD is critical to the suppression of Ci‐AChE in aphidicolin‐treated embryos.
Hepatitis C virus prevalence and outcomes among injecting drug users on opioid replacement therapy
Authors: HALLINAN RICHARD , BYRNE ANDREW , AMIN JANAKI , DORE GREGORY J
Published: August 2010

Objectives: To determine hepatitis C virus (HCV) prevalence among injecting drug users (IDUs) receiving opioid replacement therapy in a referred office setting, and assess potential needs for hepatitis C treatment and care.

Methods: Data were collected on 178 IDUs receiving opioid replacement therapy who underwent clinical assessment between January 2002 and June 2003. Standard clinic protocols included HCV and hepatitis B virus (HBV) serology, liver biochemistry and HCV RNA analysis for patients with a positive HCV antibody and normal alanine aminotransferase (ALT) levels.

Results: HCV prevalence was 75.3%, similar for males (75.5%) and females (74.4%), and increased with age from 60.8% for 19–30 years to 93.9% above 40 years. Among patients with HCV antibodies and no prior HCV antiviral therapy (n = 130), 53.1% had normal ALT levels and 25.4% were HCV‐RNA negative. Older patients were more likely to have normal ALT levels (P = 0.02), and be HCV‐RNA negative (P = 0.02). Younger patients were more likely to have been HBV vaccinated (P < 0.001), however, were less likely to have either vaccine or natural immunity (P = 0.006). Of 97 patients with probable chronic HCV infection, 58 patients met pre‐liver biopsy criteria for HCV treatment, 34 had relative contraindications to treatment and 6 had been referred for treatment assessment.

Conclusion: Clinical characterization in a setting of high HCV prevalence has enabled the differentiation of patients into groups with no evidence of HCV viraemia, with chronic HCV infection, and those most appropriate for HCV treatment referral. These clinical assessments along with appropriate referral should be instituted in drug dependency treatment settings.
Outbreak of hepatitis C virus infection in an outpatient clinic
Authors: ISHIKAWA TAKASHI , FUKUSHIMA YASUSHI , SHIOBARA YUJIRO , KISHIMOTO TSUYOSHI , TANNO SAKIKO , SHOJI IKUO , SUZUKI TETSURO , MATSUI TAMANO , SHIMADA YASUSHI , OHYAMA TAKAAKI , NAGAI RYOZO , MIYAMURA TATSUO
Published: August 2010

Background:  From January through September 2001, seven patients were admitted to Fukaya Red Cross Hospital with typical clinical manifestations of acute hepatitis. Six were outpatients of the clinic, which is located near the hospital. An extensive survey of clinic outpatients conducted by the local health department revealed six more new acute hepatitis cases during this period.

Methods:  A case control study was carried out to identify potential risk factors for infection. In total, 1946 outpatients with clinic records were scheduled to undergo hepatitis C virus (HCV)‐antibody testing. For the HCV‐Ab positive patients, HCV‐RNA was subtyped and quantified, and sequences of HCV hypervariable region 1 were determined.

Results:  Ultimately, 12 patients with acute hepatitis and two asymptomatic subjects were found to be a part of this outbreak. HCV isolates were divided into three major groups using phylogenetic tree analysis. Only a past history of visiting the clinic was significantly associated with acute hepatitis. The timing of the parenteral medical procedure at the clinic and the onset of acute hepatitis strongly suggested association of the two events.

Conclusions:  Our findings suggest that nosocomial HCV infection can occur in an outpatient clinic, even in countries where post‐transfusion hepatitis has been almost entirely eliminated.
Hydroxyproline content of needle biopsies as an objective measure of liver fibrosis: Emphasis on sampling variability
Authors: LEE HSUAN‐SHU , SHUN CHIA‐TUNG , CHIOU LING‐LING , CHEN CHIEN‐HUNG , HUANG GUAN‐TARN , SHEU JIN‐CHUAN
Published: August 2010

Background:  Needle liver biopsy is essential for the diagnosis of liver fibrosis and cirrhosis. However, sampling variability has been reported by semiquantitative methods. The aim of the present study was to investigate quantitatively the sampling variability of needle liver biopsy on the assessment of liver fibrosis/cirrhosis.

Methods:  Liver samples of 12 patients undergoing autopsy, liver transplantation, or liver tumor resection were investigated. Nine to 10 liver biopsies at least 5 mm apart were taken from each of 12 removed human livers using Trucut needles. Hydroxyproline level of each liver core was determined. Sections >1 cm2 and stained by Sirius red were used for the reference staging of liver fibrosis in each patient according to the METAVIR scoring system.

Results:  Hydroxyproline levels for the three groups, F0‐1, F2‐3, and F4, were significantly elevated as the fibrosis score increased. There was a wide range of hydroxyproline contents for the 9–10 biopsy samples from each liver, the coefficient of variation being between 13% and 36% (mean, 23%). Several differences were noted between the amount of collagen present according to hydroxyproline measurement and the stages of fibrosis according to histology.

Conclusions:  Although increases in collagen content (by hydroxyproline) approximately paralleled increases in fibrosis stage (by histology), there was considerable overlap and some discrepancies. Much work remains to be done to clarify the significance of the apparent huge variations in fibrosis within diseased livers. It is advisable for clinicians to interpret staging diagnoses by histological staging with some caution.
Effects of somatostatin, terlipressin and somatostatin plus terlipressin on portal and systemic hemodynamics and renal sodium excretion in patients with cirrhosis
Authors: KALAMBOKIS GEORGIOS , ECONOMOU MICHALIS , PARASKEVI KOSTA , KONSTANTINOS PAPADIMITRIOU , PAPPAS CHRISTOS , KATSARAKI AFRODITI , TSIANOS EPAMEINONDAS V
Published: August 2010

Background and Aim:  Terlipressin and somatostatin are the most preferable agents for the control of variceal bleeding in cirrhotic patients. The present study evaluated the hemodynamic effects of somatostatin, terlipressin and somatostatin plus terlipressin in cirrhotic patients with portal hypertension, as well as the effect of each regimen on renal sodium excretion.

Methods:  Twenty‐four patients with esophageal varices were randomly assigned to receive either an intravenous infusion of a placebo (n = 12) or somatostatin 250 µg/h after an initial bolus of 250 µg (n = 12) for 60 min. Thereafter, each patient received an intravenous injection of terlipressin 2 mg while the intravenous infusion of either somatostatin or placebo was maintained. Portal and systemic hemodynamic parameters, assessed by Doppler sonography, and urinary sodium excretion were evaluated at baseline, 60 min after placebo or somatostatin, and 30 min after terlipressin.

Results:  Placebo had no effect on the patients studied. After terlipressin, portal vein velocity, portal flow volume and cardiac output (CO) significantly decreased (0.09 vs 0.15 m/s, 0.56 vs 1 L/min and 6.4 vs 7.6 L/min, respectively [values are medians]), while mean arterial pressure (MAP) and systemic vascular resistance significantly increased (103.3 vs 89.9 mmHg and 1541 vs 1108dyn.s/cm5, respectively). Fractional sodium excretion significantly increased in patients without ascites (0.43 vs 0.16%) while it did not change in patients with ascites. Somatostatin did not alter portal hemodynamics whereas it significantly reduced MAP, heart rate (HR) and CO (86.9 vs 98.6 mmHg, 65 vs 73 bpm and 8.4 vs 9.1 L/min, respectively) and, in patients with ascites, sodium excretion (0.13 vs 0.23%). The addition of terlipressin to somatostatin induced similar changes to those observed after terlipressin alone. The magnitude of increase in MAP was significantly higher in patients receiving terlipressin alone than in those receiving somatostatin plus terlipressin (15 vs 5.3%), while CO was conversely affected (−28.5 vs−20.9%).

Conclusions:  Combined treatment with somatostatin and terlipressin does not exert an additive portal hypotensive effect in cirrhotic patients as compared to terlipressin alone, whereas somatostatin alone may impair systemic hemodynamics. Compared with somatostatin, terlipressin exerts a more beneficial effect on renal sodium excretion in patients with or without ascites.
Aberrant expression and localization of hnRNP‐A2/B1 is a common event in human gastric adenocarcinoma
Authors: Jing Guang‐Jun , Xu Dong‐Hui , Shi Song‐Lin , Li Qi‐Fu , Wang San‐Ying , Wu Fu‐Yun , Kong Hai‐Yan
Published: August 2010

Background and Aim:  Nuclear‐matrix proteins can be proteomic markers for cancer lesions. The present study aimed to determine the roles of heterogeneous nuclear ribonucleoproteins‐A2 and B1 (hnRNP‐A2/B1) in human gastric carcinogenesis.

Methods:  Human gastric cancer and non‐cancerous tissues were collected for immunohistochemical analysis. Proteomics technique, Western blot, laser confocal microscope, and real‐time quantitative reverse transcription–polymerase chain reaction were performed to determine the aberrant expression of nuclear‐matrix proteins.

Results:  hnRNP‐A2/B1 existed in the nuclear matrix of gastric cancer cells, and its expression was enhanced in human gastric cancer and decreased by hexamethylene bisacetamide. The colocalization of hnRNP‐A2/B1 with c‐myc, c‐fos, p53, and Rb was translocated from the nucleolus to the cytoplasm during the differentiation of tumor cells.

Conclusions:  hnRNP‐A2/B1 affected tumor cell differentiation through interaction with oncogenes and tumor‐suppressor genes, and it was overexpressed in human gastric cancer. We postulate that hnRNP‐A2/B1 could serve as a biomarker for the diagnosis of human gastric cancer.
Analysis of K‐ras, BRAF, and PIK3CA mutations in laterally‐spreading tumors of the colorectum
Authors: Kaji Eisuke , Kato Jun , Suzuki Hideyuki , Akita Mitsuhiro , Horii Joichiro , Saito Shunsuke , Higashi Reiji , Ishikawa Shin , Kuriyama Motoaki , Hiraoka Sakiko , Uraoka Toshio , Yamamoto Kazuhide
Published: August 2010

Background and Aims:  Laterally‐spreading tumors (LST) are a newly‐recognized category of colorectal neoplasia, and are defined as lesions larger than 10 mm in diameter and extending circumferentially rather than vertically. However, genetic features of this new category of tumors are not fully elucidated. The aim of this study was to evaluate genetic alterations in LST.

Methods:  We examined K‐ras, BRAF, and phosphoinositide‐3‐kinase catalytic‐α polypeptide (PIK3CA) mutations in 101 LST, including 68 LST‐granular type (LST‐G) and 33 LST‐non‐granular type by direct sequencing. As controls, we examined these gene mutations in 66 protruded colon adenomas (10 mm or larger) and 44 advanced colon cancers.

Results:  K‐ras, BRAF, and PIK3CA mutations were observed in 59 (58%), zero (0%), and three (3%) LST, respectively. LST‐G morphology in the right‐sided colon was significantly correlated with the existence of K‐ras mutations, whereas a size of 20 mm or larger was the only predictor of mutations in the left‐sided colorectum. The frequency of K‐ras mutations in LST was particularly marked in the left‐sided colorectum compared to protruded adenomas or advanced cancers (LST vs protruded adenomas, P < 0.001; LST vs advanced cancers, P = 0.002), whereas in the right‐sided colon, K‐ras mutations were equally frequent. PIK3CA mutations were not familiar in either LST (3%) or advanced cancers (9%).

Conclusions:  K‐ras mutations were involved in colorectal LST in different manners according to tumor location.
Controversies of cardiac glands in the proximal stomach: A critical review
Authors: Huang Qin
Published: August 2010

Cardiac glands (CG), along with oxyntocardiac glands, in a normal human constitute cardiac mucosa (CM) that is positioned in the proximal stomach with a length of 10–30 mm, according to traditional teaching. This doctrine has been recently challenged. On the basis of studies on autopsy and biopsy materials in the esophagogastric junction region, some investigators have reported the presence of CG in only 50% of the general US population. They believed that CG were an acquired, metaplastic lesion as a result of gastroesophageal reflux disease. Subsequent recent study results from other research groups showed the presence of CG in the proximal stomach in embryos, fetuses, pediatric, and adult patients in most Europeans and Americans, and almost all Japanese and Chinese patients. These new data showed the following important findings: (i) CG are confirmed to be congenital in the proximal stomach; (ii) the length of CM is much shorter, approximately 5 mm in Caucasians in Europe and North America, and approximately 13 mm in Japanese and probably also in Chinese; (iii) CG are also present in the distal superficial esophagus underneath squamous mucosa in almost all Japanese and Chinese patients, but not so common in Caucasians in Europe, and not clear in Caucasians in North America. The recent data indicate a clear difference in the distribution of CG in the proximal stomach among different ethnic populations, and might explain different disease pathogenesis mechanisms among various ethnic patient groups.
Treatment of hepatitis C virus infection in patients with end‐stage renal disease
Authors: Liu Chen‐Hua , Kao Jia‐Horng
Published: August 2010

Hepatitis C virus (HCV) infection is a major health problem in patients with end‐stage renal disease (ESRD). The incidence of acute HCV infection during maintenance dialysis is much higher than that in the general population because of the risk of nosocomial transmission. Following acute HCV infection, most patients develop chronic HCV infection, and a significant proportion develop chronic hepatitis, cirrhosis, and even hepatocellular carcinoma. Overall, chronic hepatitis C patients on hemodialysis bear an increased risk of liver‐related morbidity and mortality, either during dialysis or after renal transplantation. Interferon (IFN) therapy is modestly effective for the treatment of HCV infection in ESRD patients. Conventional or pegylated IFN monotherapy has been used to treat acute hepatitis C in ESRD patients with excellent safety and efficacy. Regarding chronic hepatitis C, approximately one‐third of patients can achieve a sustained virological response (SVR) after conventional or pegylated IFN monotherapy. The combination of low‐dose ribavirin and conventional or pegylated IFN has further improved the SVR rate in treatment‐naïve or retreated ESRD patients in clinical trials. Similar to the treatment of patients with normal renal function, baseline and on‐treatment HCV virokinetics are useful to guide optimized therapy in ESRD patients. Of particular note, IFN‐based therapy is not recommended at the post‐renal transplantation stage because of the low SVR rate and risk of acute graft rejection. In conclusion, ESRD patients with HCV infection should be encouraged to receive antiviral therapy, and those who achieve an SVR usually have long‐term, durable, virological, biochemical, and histological responses.
Allopurinol might improve response to azathioprine and 6‐mercaptopurine by correcting an unfavorable metabolite ratio
Authors: Gardiner Sharon J , Gearry Richard B , Burt Michael J , Chalmers‐Watson Teresa , Chapman Bruce A , Ross Alison G , Stedman Catherine A M , Huelsen Alexander , Barclay Murray L
Published: August 2010

Background and Aim:  Allopurinol potentiates azathioprine and 6‐mercaptopurine (6‐MP) by increasing 6‐thioguanine nucleotide (6‐TGN) metabolite concentrations. The outcome might also be improved by adding allopurinol in individuals who preferentially produce 6‐methylmercaptopurine nucleotides (6‐MMPN), rather than 6‐TGN. The aim of the present study was to investigate the effect of allopurinol on concentrations of 6‐MMPN and 6‐TGN in individuals with a high ratio of these metabolites (>20), which is indicative of a poor thiopurine response.

Methods:  Sixteen individuals were identified who were taking azathioprine or 6‐MP, and were commenced on allopurinol to improve a high 6‐MMPN : TGN ratio. Metabolite concentrations were compared before and after commencing allopurinol, and markers of disease control were compared.

Results:  The addition of 100–300 mg allopurinol daily and thiopurine dose reduction (17–50% of the original dose) resulted in a reduction of the median (and range) 6‐MMPN concentration, from 11 643 (3 365–27 832) to 221 (55–844) pmol/8 × 108 red blood cells (RBC; P = 0.0005), increased 6‐TGN from 162 (125–300) to 332 (135–923) pmol/8 × 108 RBC (P = 0.0005), and reduced the 6‐MMPN : 6‐TGN ratio from 63 (12–199) to 1 (0.1–4.5) (P = 0.0005). There was a significant reduction in steroid dose requirements at 12 months (P = 0.04) and trends for improvement in other markers of disease control. One patient developed red cell aplasia that resolved upon stopping azathioprine and allopurinol.

Conclusions:  In those with a high 6‐MMPN : 6‐TGN ratio (>20), response to thiopurine treatment might be improved by the addition of allopurinol, together with a reduced thiopurine dose and close hematological monitoring.
PROBLEMS IN THE MANAGEMENT OF NEONATAL MENINGITIS
Authors: ROBINSON M. J.
Published: August 2010

SYNOPSIS

Undoubtedly much of the morbidity and mortality associated with neonatal meningitis is related to delay in diagnosis. Undoubtedly also, organisms such as proteus and pyocyaneus are more lethal. Despite this, there is adequate evidence that electrolyte imbalance from inappropriate A.D.H. secretion can compound the problems of the meningitis and thus disturb cerebral function. Careful monitoring of the serum sodium and also appropriate fluid restriction will prevent this. In addition, there should be an awareness of the frequency of ventriculitis in this age group. Ventriculitis can only be effectively managed by the local instillation of chemotherapeutic agents and a Rickham reservoir is a most satisfactory means of accomplishing this end.
THE INFANT BORN OF A SINGLE MOTHER PREGNANCY A STUDY OF THE RISK
Authors: JOHNS NAN
Published: August 2010

SYNOPSIS

The pregnancy and delivery histories of two groups of single mothers confined in a Melbourne public maternity hospital in 1968 were compared with those of a control group of married women. No significant incidence of factors liable to influence adversely the well‐being of the infants born to the single mothers was found.
PREDICTION OF SURVIVAL OF VERY LOW BIRTH WEIGHT INFANTS
Authors: KITCHEN W. H. , GAUDRY E.
Published: August 2010

SYNOPSIS

A method has been developed of predicting survival to 28 days of 238 consecutive births weighing between 1000 and 1500 g. Sixteen possible predictors of neonatal death were used for discriminant function and multiple regression analyses
HYPERCALCIURIA IN POTASSIUM‐LOSING NEPHROPATHY:
Authors: McCREDIE D. A. , ROTENBERG E. , WILLIAMS A. L.
Published: August 2010

SYNOPSIS

The case histories of 4 children with Bartter's syndrome are presented. All were born prematurely and pregnancy was associated with hydramnios. All failed to thrive and had a vasopressin‐insensitive urinary concentrating defect, but were of normal intelligence. Hypo‐kalaemia was not necessarily present when first seen, but serum sodium was low compared with other patients with diabetes insipidus. Salt loading caused a sharp fall in serum potassium, often associated with a paradoxical rise in plasma renin levels. Urine calcium was high in all patients and rose to very high levels with salt loading. Nephrocalcinosis was a constant finding. The basic defect is thought to be one of ion absorption in the proximal tubule, though membrane transport may also be affected in other areas.
HYPERTENSION IN CHILDHOOD OBESITY
Authors: COURT J. M. , HILL G. J. , DUNLOP MARJORIE , BOULTON T. J. C.
Published: August 2010

SYNOPSIS

Blood pressure has been measured in 209 children on first consultation when referred to the Obesity Research Clinic. Blood pressure correlated with estimates of obesity made by subscapular‐skinfold thickness, and calculations of per cent body weight as fat. Neither age nor duration of the obesity showed significant correlation with the blood pressure.
Is prescribing paracetamol ‘pro re nata’ acceptable?
Authors: PENNA A. C. , DAWSON K. P. , PENNA C. M.
Published: August 2010

Abstract The aim of this study was to assess the prescribing pattern of paracetamol in a children's department of a general hospital. A retrospective audit of the case record prescriptions revealed that 190 of a sample of 299 children had received paracetamol during their inpatient stay.

The major indications for paracetamol use were fever associated with infection in medical patients and pain following minor surgical procedures. The most common regimen was 4 hourly p.r.n. and this had the greatest potential to lead to a dose in excess of 90 mg/kg per 24h, other than the actual prescribing of high doses on a fixed basis. This dose was actually administered to 13.6% of the children, and was prescribed for 74% of the group. We conclude that a p.r.n. regimen can lead to overdose and this may be potentially hazardous for children.
Individualizing aminophylline doses in premature infants using bioelectrical impedance: A non‐invasive approach
Authors: SIDHU J. , TRIGGS E. , CHARLES B. , TUDEHOPE D. , GRAY P. , COWLEY D.
Published: August 2010

Abstract The role of bioelectrical impedance (BI) in estimating the pharmacokinetics and, therefore, individualized doses, of aminophylline in preterm infants (gestational age 26–35 weeks) was assessed in a two‐phase study. Multiple regression analysis in the first group of neonates (phase I, n= 19) identified resistance, reactance, weight and length as optimal predictors of distribution volume (adjusted R2= 0.84, coefficient of variation (CV) = 10.17%), and length2/impedance and postconceptual age as predictors of clearance (adjusted R2= 0.74, CV = 26.73%). Application of these models to an independent group (phase II, n= 20) generated doses which satisfactorily achieved target theophylline loading and steadystate (SS) ‘peaks’ of 10 μg/mL and SS ‘troughs’ of 7.7±0.6 μg/mL. A better understanding of specific criteria and limitations of the impedance technique in neonates is necessary in order to refine BI measurements.
Ophthalmic assessment of physically disabled children attending a rehabilitation centre
Authors: FLETT P. , SAUNDERS B.
Published: August 2010

Abstract A retrospective analysis was conducted of 397 children, ranging in age from 2 to 19 years, attending the Regency Park Centre for Young Disabled in Adelaide. The disorders represented included cerebral palsy, head injury, spina bifida, severe speech and/or language disorders, muscular dystrophy and a number of less common conditions. The incidence of significant ophthalmic abnormalities in this population was 51%, with the highest incidence being 69 and 62% among children with head injury and cerebral palsy, respectively. Children with severe speech and language disorders had an incidence of 24%. Overall, routine examinations revealed previously unsuspected significant eye abnormalities in 31% of the children. A complete ophthalmic examination should be part of the evaluation of all physically disabled children to ensure early identification and treatment of abnormalities, and to optimize rehabilitation.
Childhood injury mortality in New South Wales: Geographical and socio‐economic variations
Authors: CAREY V. , VIMPANI G. , TAYLOR R.
Published: August 2010

Abstract This paper reports a retrospective study of deaths due to unintentional injury in children aged between 0 and 14 years of age in New South Wales between 1985 and 1987. The aims were to determine whether the pattern of child injury mortality differed between rural and metropolitan New South Wales and to establish the relationship between socio‐economic status, based on a geographical indicator of socio‐economic status and child injury mortality.

The child injury mortality rate for New South Wales in 1985–87 was 12.3 deaths per 100000 population per year. The rate in the country area of 15.3 deaths per 100000 was significantly higher than that for the metropolitan area of 11.3 per 100000 (P<0.05). Deaths of passengers in motor vehicle traffic accidents (P<0.01) and deaths due to fires (P<0.01) were greater in the country area. A negative linear association between socio‐economic status and child injury mortality was found in the Sydney metropolitan area (P<0.01). This trend was most apparent for deaths of child pedestrians (P<0.01).