bluuebunny/crossref_metadata_2025 huggingface dataset:
Internalization and Propagation of the Dengue Virus in Human Hepatoma (HepG2) Cells
Authors: Thepparit Chutima , Phoolcharoen Waranyoo , Suksanpaisan Lukkana , Smith Duncan R.
Published: June 2004

Objectives: This study sought to undertake a comparative analysis of the internalization and propagation of all four dengue serotypes in a single cell line of human liver origin, HepG2. Methods: Virus production after infection was determined by the plaque assay technique. Internalization profiles were determined by incubating virus and cells on ice and then raising the temperature for various times. The contribution of extracellular matrix components to internalization was determined by pretreatment of cells with either trypsin or heparinase III. Results: HepG2 cells were able to support the propagation of all four serotypes with mature viruses being produced by 12 h for dengue serotype 4 and by 17–18 h for the remaining serotypes. Virus internalization showed a plateau for serotypes 1, 2 and 4 entry while serotype 3 showed a constant increase in internalization for up to 5 h. Pretreatment of HepG2 cells with heparinase III or trypsin both resulted in a reduction in viral production, with the smallest effect being noted for dengue serotype 3. Conclusion: These results suggest that the interaction between the dengue virus and liver cells is a complex one that requires both protein and nonprotein elements, and has a significant serotype/strain element.
Establishing the Reactivity of Monoclonal Antibodies against Porcine Endogenous Retrovirus Envelope Protein
Authors: Wang Shih-Rong , Chang Jen-Ting , Lin Meng-Shiue , Chiang Chen-Yi , Chang Hwan-You
Published: June 2004

Xenotransplantation of pig organs may be associated with a risk of transmission of microorganisms. Porcine endogenous retroviruses (PERV) are of particular concern since in vitro experiments have demonstrated that human cells are susceptible to such microorganisms. To monitor the transmission of PERV, highly sensitive and specific immunoassays must be developed for clinical surveillance. This report describes the production, preliminary characterization and application of a monoclonal antibody (mAb) against a recombinant PERV envelope (Env) protein. The generated mAb was tested using recombinant PERV Env protein expressed in Escherichia coli, purified PERV virus particles and human 293 cell line infected with PERV. PERV-translated proteins of 15, 70 and 85 kD were recognized specifically using PERV-8E10 mAb and Western blotting. No cross-reactivity was demonstrated with exogenous viral protein (HIV, HTLV and MuLV). Moreover, PERV-8E10 mAb can be applied to localize PERV proteins using an immunoperoxidase assay. This work reveals that recombinant PERV Env protein and mAb may be effective in detecting antibodies against PERV in xenotransplanted patients, or for butchers who have extensive contact with pigs.
A Comparison of the Exponential Decay Slope between PEG-IFN alfa-2b/Ribavirin and IFN alfa-2b/Ribavirin Combination Therapy in Patients with Chronic Hepatitis C Genotype 1b Infection and a High Viral Load
Authors: Izumi Namiki , Asahina Yasuhiro , Kurosaki Masayuki , Uchihara Masakatsu , Nishimura Yuki , Inoue Kazunari , Ueda Ken , Tsuchiya Kaoru , Hamano Kousei , Itakura Jun , Miyake Shozo
Published: June 2004

Objectives: A high virological response rate can often be shown to be obtained with PEG-IFN α-2b and ribavirin combination therapy in chronic hepatitis C patients. Viral dynamics have been utilized for the evaluation of antiviral effects, especially the exponential second decay slope, which represents the elimination of infected cells. Methods: Forty-nine patients were randomly assigned to the IFN α-2b group (n = 26) or the PEG-IFN α-2b group (n = 23). Ribavirin was administered equally to both groups. Measuring the serum concentration of HCVRNA, the exponential viral decay during phase 1 and 2 was calculated. Results: The exponential decay slope in phase 2 during the first 2 weeks was greater in the IFN α-2b group than in the PEG-IFN α-2b group; however, from weeks 3 to 4, it was greater in the PEG-IFN α-2b group than in the IFN α-2b group. Interestingly, in the PEG-IFN α-2b group, the exponential decay slope was greater from weeks 3 to 4 after initiating combination therapy than during the weeks 1–2 (p < 0.01), despite administration of the same PEG-IFN α-2b dose (1.5 µg/kg once weekly). Conclusions: In PEG-IFN α-2b and ribavirin combination therapy, elimination of infected cells may be pronounced following an increase in serum ribavirin concentration in chronic hepatitis C patients with genotype 1b infection and a high viral load.
Abklärung eines auffälligen Zervixabstrichs – Braucht es dazu Dysplasiezentren?
Authors: Heinzl S.
Published: June 2004

Die Spezialisierung in der Medizin hat in den letzten Jahren zugenommen. Es werden mittlerweile verschiedene Zentren (Mamma, Fertilität etc.) eröffnet. Auch Dysplasie- (Kolposkopie-)zentren und -sprechstunden stehen zur Diskussion. Als Gründe werden das neu etablierte differenzierte Vorgehen bei CIN/SIL, VAIN und VIN, die geringere Patientinnenzahl pro Arzt und die mangelhafte Qualität angegeben. Die Konzentration auf Spezialisten würde helfen, die Qualität zu verbessern, und könnte auch helfen, Kosten zu sparen. Eine Dysplasiesprechstunde oder -zentrum kann unter bestimmten Bedingungen eine optimale Einrichtung für Patientinnen mit Erkrankungen des unteren Genitaltraktes sein.
Die Frauenärztin, der Frauenarzt als «Hausarzt der Frau» – ein Zukunftsmodell?
Authors: Kreienberg Rolf
Published: June 2004

Gynäkologie und Geburtshilfe bilden ein Fachgebiet, das Präventivmedizin, Primärversorgung der Frau und hochqualifizierte Beckenchirurgie sowie Massnahmen der Geburtshilfe und der Endokrinologie vereinigt. Die Notwendigkeit, Ressourcen zu bündeln, insbesondere bei begrenzten Budgets, erfordert ein Umdenken in der Politik, bei den Anbietern im Gesundheitssystem und bei den Gynäkologen und Geburtshelfern. Der Artikel beschreibt, wie eine Primärversorgung der Patientin im Sinne eines Hausarztes der Frau durch die niedergelassenen Frauenärztinnen und Frauenärzte erfolgen könnte und welche Anforderungen bzw. welches Leistungs- und Ausbildungsspektrum dafür erforderlich sind.
Reduction of Pleural Effusion by OK-432 in a Fetus Complicated with Congenital Hydrothorax
Authors: Tsukihara Ayumi , Tanemura Mitsuyo , Suzuki Yoshikatsu , Sato Takeshi , Tanaka Taihei , Suzumori Kaoru
Published: June 2004

A 29-year-old, primiparous woman was referred to our hospital at 21 weeks of gestation because of right pleural effusion in the fetus shown by routine ultrasonographic examination. Cytology revealed abundant lymphocytes, suggesting chylothorax. We removed the pleural effusion and injected OK-432 into the chest cavity at 24 and 25 weeks of gestation. Pleural effusion declined and an adhesion between the lung surface and the pleural membrane seemed to form. At 33 weeks of gestation, a female infant was born by cesarean section (1,090 g and Apgar score 6/8). Although she demonstrated slight retraction and tachypnea, management could be achieved by administration of oxygen alone without mechanical ventilation. Later, the baby was diagnosed as suffering from the Cornelia de Lange syndrome with characteristic features. OK-432 injections could thus prevent complications of chylothorax and hypoplastic lungs, without injury to either the baby or the mother.
Dandy-Walker Malformation: A Review of 78 Cases Diagnosed by Prenatal Sonography
Authors: Has Recep , Ermiş Hayri , Yüksel Atıl , Ibrahimoğlu Lem’i , Yildirim Alkan , Sezer Hande Delier , Başaran Seher
Published: June 2004

Objective: The purpose of this study was to determine the associated abnormalities and clinical outcomes of fetuses with Dandy-Walker malformations. Methods: Sonograms and medical reports of 78 cases were reviewed and information regarding each outcome was collected from autopsy records, hospital charts and specialists caring for the surviving infants. Results: We identified 64 fetuses with classic Dandy-Walker malformation (DW) and 14 fetuses with Dandy-Walker variant (DWV). A high proportion (44.8%) of the parents were consanguineous. The spectrum and proportion of central nervous system (67.1 vs. 71.4%; p = 1.0) and other malformations (43.7 vs. 64.2%; p = 0.57) associated with DW and DWV were similar. Chromosome abnormalities were found in 9 of the 51 (17.6%) fetuses that underwent karyotype analysis. Only 4 of 64 (6.2%) DW and 3 of 14 (21.4%) DWV infants survived (p 0.14), and all surviving infants with DW or DWV had neurological disorders. Conclusions: DW and DWV cases show so many similarities that a clear-cut distinction is difficult. There was no significant difference in the spectrum of associated anomalies and postnatal prognosis between DW and DWV cases.
Importance of Ductus venosus Doppler Assessment for Fetal Outcome in Cases of Intrauterine Growth Restriction
Authors: Ritter Stefan , Jörn Hendrik , Weiss Claudia , Rath Werner
Published: June 2004

Objective: The measurement and evaluation of ductus venosus (DV) blood flow velocity waveform in high-risk pregnancies has been studied intensively in recent years in order to find a more intermediate signal of fetal compromise. Our objective was to study the fetal outcome of pregnancies with intrauterine growth retardation (IUGR) and normal pulsatility of DV compared to an IUGR group with increased DV pulsatility. Methods: The outcome of 42 fetuses before 32 weeks of gestational age without chromosomal or structural aberrations was analyzed. All fetuses showed IUGR <5th percentile based on placental insufficiency diagnosed by pathologic RI >90th percentile of both maternal uterine arteries. One group (30 fetuses; mean weight 730 g/SD 190 g; mean gestational age 197 days/SD 12 days) showed normal, the other (12 fetuses, mean weight 675 g/SD 179 g; mean gestational age 198 days/SD 12 days) reduced, but neither absent nor reverse DV flow during atrial contraction. All 42 fetuses were delivered by cesarean section because of severe variable or prolonged decelerations. We measured blood flow velocities of the DV in every fetus on an average 3.7 days (range 1–5 days) before cesarean section. Fetal outcome was determined by Apgar scores after 5 and 10 min, arterial pH and base excess; neonatal morbidity was recorded by intensive follow-up. Results: There were no significant differences of pH, umbilical artery base excess, Apgar scores and severe neonatal complications between the two groups. Conclusions: Our own data show no correlation between increased pulsatility in the DV (without absent or reverse flow during atrial contraction) and fetal outcome before 32 gestational weeks, even in cases of severe growth restriction based on placental insufficiency. Therefore in these cases reduced DV flow during atrial contraction should cautiously be interpreted regarding obstetrical decisions.
Prenatal Diagnosis of a Fetus Affected with Down Syndrome and Deletion 1p36 Syndrome by Fluorescence in situ Hybridization and Spectral Karyotyping
Authors: Hsieh Lie-Jiau , Hsieh Tsung-Che , Yeh Guang-Perng , Lin Meng-I , Chen Ming , Wang Boris Bao-Tyan
Published: June 2004

Objective: A fetus having partial trisomy of the distal part of chromosome 21q due to a de novo translocation is reported here. Method: A 29-year-old woman received amniocentesis at 18 weeks of gestation because of abnormal ultrasound findings including bilateral choroid plexus cysts, atrioventricular septal defects, rocker-bottom feet, and possible hydrocephalus. Results: Cytogenetic analysis revealed 46,XY, add(1)(p36.3), in which an additional material of unknown origin was attached to one of the terminal short arms of chromosome 1. Parental blood studies showed normal karyotypes in both parents. Spectral karyotyping was then performed and the origin of the additional material locating at chromosome 1p was found to be from chromosome 21. Conventional fluorescence in situ hybridization analysis was also used and confirmed the spectral karyotyping findings by use of a chromosome 21 specific painting probe, a locus specific probe localized within bands 21q22.13–q22.2 and a 21q subtelomeric probe. A hidden Down syndrome caused by a de novo translocation in this fetus was therefore diagnosed and the karyotype was designated as 46,XY, der(1)t(1;21)(p36.3;q22.1).ish der(1)(WCP21+, LSI 21+, 1pTEL–, 21q TEL+) de novo. Clinical features of the 1p36 deletion syndrome are also reviewed and may contribute to some features of this fetus. Termination of pregnancy was performed at 20 weeks of gestation. Conclusion: To our knowledge, our case appears to be the first to have partial monosomy 1p and partial trisomy 21q caused by de novo translocation being diagnosed prenatally.
Antenatal Necrotic Injury in Severe Twin-to-Twin Transfusion Syndrome
Authors: Carr Stephen R. , Luks François , Tracy Thomas , Plevyak Michael
Published: June 2004

A case of twin-to-twin transfusion syndrome (TTTS) was associated with ischemic injury of the left leg of the recipient twin. Five reduction amniocenteses were performed between 17 6/7 and 23 3/7 weeks followed by fetoscopic laser ablation of chorioangiopagus vessels at 23 6/7 weeks of gestation. Anatomic assessment of the affected limb indicated timing of the injury at 19 4/7 weeks gestational age. The published literature suggests that ischemic fetal injuries in severe TTTS are associated with blood hyperviscosity in the affected twin.
Congenital Laryngeal Cyst: Benefits of Prenatal Diagnosis and Multidisciplinary Perinatal Management
Authors: Busuttil M. , Einaudi M.-A. , Hassid S. , Chau C. , Shojai R. , Thomachot L. , D’Ercole C. , Gire C.
Published: June 2004

Congenital laryngeal cysts are rare and can cause upper airway obstruction if they are misdiagnosed. We describe a case of large vallecular cyst diagnosed at 33 weeks of gestation. The purpose of an early diagnosis is to establish a careful perinatal management. If the airway is compromised, a multidisciplinary medical team is necessary to perform emergency immediate tracheostomy. Elective delivery should be always carried out in a tertiary referral center.
Prospective Evaluation of Uterine Artery Flow Velocity Waveforms at 12–14 and 22–24 Weeks of Gestation in Relation to Pregnancy Outcome and Birth Weight
Authors: Carbillon L. , Uzan M. , Largillière C. , Perrot N. , Tigaizin A. , Paries J. , Pharizien I. , Uzan S.
Published: June 2004

Objective: Uterine artery flow velocity was prospectively assessed using Doppler ultrasound at 12–14 and 22–24 weeks of gestation in the prediction of subsequent complications related to uteroplacental insufficiency: preeclampsia, pregnancy-induced hypertension, fetal growth restriction, fetal death and placental abruption, and to elucidate its relationship with birth weight. Methods: Uterine artery Doppler assessment was obtained during routine ultrasound screening in 263 unselected women. Flow velocity waveforms were coded according to the number of notches present at each scanning, respectively: none (0, 0), uni-/bilateral notches that disappeared (1, 0) or (2, 0), uni-/bilateral notches that persisted unilaterally (1, 1) or (2, 1), and persistent bilateral notches (2, 2). Results: Complete outcome data was obtained for 243 (92.4%) women. Of these women, 55 (22.6%) and 84 (34.6%) women had uni- and bilateral notches, respectively, at 12–14 weeks’ gestation; 14 (5.8%) and 21 (8.6%) patients had uni- and bilateral notches, respectively, at 22–24 weeks’ gestation. Analysis of complication rates for the four groups showed that they increased with notch persistence (5.7, 13.5, 57.1 and 76.2%), while the corresponding mean birth weight declined (3,273, 3,180, 2,698 and 2,418 g). Conclusion: The absence or early disappearance of uterine artery notches is associated with fewer complications related to uteroplacental insufficiency and normal birth weight, whereas their late and partial disappearance or bilateral persistence tends to compromise the prognosis.
Surfactant Protein B: Structure and Function
Authors: Hawgood Sam
Published: June 2004

Pulmonary surfactant is a mixture of phospholipids, neutral lipids, and associated proteins. A specific phospholipid, dipalmitoylphosphatidylcholine, is predominantly responsible for the modulation of surface tension at the alveolar air-liquid interface, but other surfactant lipid and protein components play important roles in surfactant function and metabolism. This review will focus on just one of the apoproteins, surfactant protein B, with a description of protein structure and the actions of surfactant protein B on surfactant lipid membranes.
Plasma Membrane Stress Failure in Ventilator-Injured Lungs
Authors: Mendez Jose L. , Rickman Otis B. , Hubmayr Rolf D.
Published: June 2004

Cell injury and repair are invariable consequences of mechanical ventilation with large tidal volumes. Rate and amplitude of deforming stress affect numerous cell metabolic functions including host defense and wound repair. Recently, we have focused on the role of plasma membrane stress failure as a trigger for a pro-inflammatory response in mechanically ventilated lungs. We have developed both cell- and organ-based models to study this problem. Alveolar epithelial cells that are exposed to deforming stresses seek to maintain sublytic plasma membrane tension and may activate mechanisms of cell surface area regulation to control membrane tension. Interventions which either increase the amount of excess plasma membrane or enhance lipid trafficking should be cytoprotective against deformation induced injury. Osmotic manipulation may be one such intervention. Preconditioning the lungs with anisosmotic solutions may allow the cells to recruit excess plasma membrane and thus be more resistant to ventilator-induced lung injury.
Genetic Mechanisms of Surfactant Deficiency
Authors: Nogee Lawrence M.
Published: June 2004

The production of pulmonary surfactant is necessary to maintain alveolar stability and normal lung function. Mutations in three different genes important for surfactant production and function have now been recognized to result in surfactant deficiency and acute and/or chronic lung disease. The clinical and laboratory features associated with these genetic disorders, along with their implications for the understanding of normal surfactant metabolism, are reviewed.
Lung and Brain Damage in Preterm Newborns
Authors: Dammann Olaf , Leviton Alan , Bartels Dorothee B. , Dammann Christiane E.L.
Published: June 2004

The relationships among bronchopulmonary dysplasia (BPD), brain white matter damage (WMD) and cerebral palsy (CP) are far from simple. Apparently, BPD and WMD are not associated, while BPD and CP are. The most likely explanation for this paradox is that ultrasound imaging does not identify all the WMD that might lead to CP (‘tip-of-the-iceberg effect’). We discuss further methodological inconsistencies, etiological peculiarities related to antenatal infection/inflammation, and intervention-related issues. In particular, we expand on the multiple-hit scenario in the etiology of BPD and offer support for the hypothesis that it is not lung disease, but factors associated with lung disease (e.g. postnatal steroid exposure) that increase the risk for developmental disability in childhood.
Candidacy for the Bone-Anchored Hearing Aid
Authors: Snik Ad F.M. , Bosman Arjan J. , Mylanus Emmanuel A.M. , Cremers Cor W.R.J.
Published: June 2004

The BAHA (bone-anchored hearing aid) is a bone conduction hearing aid with percutaneous transmission of sound vibrations to the skull. The device has been thoroughly evaluated by various implant groups. These studies showed that, in audiological terms, the BAHA is superior to conventional bone conduction devices. In comparison with air conduction devices, the results are ambiguous. However, a positive effect is found with respect to aural discharge. The most powerful BAHA can be applied to patients with a sensorineural hearing loss component of up to 60 dB HL. It was shown that bilateral BAHA application leads to binaural sound processing. Preliminary results on the application of the BAHA in patients with unilateral conductive hearing loss suggest that stereophonic hearing can be re-established. The application of the BAHA as a transcranial CROS (contralateral routing of signal) device in unilateral deafness minimizes head shadow effects.
HiResolutionTM and Conventional Sound Processing in the HiResolutionTM Bionic Ear: Using Appropriate Outcome Measures to Assess Speech Recognition Ability
Authors: Koch Dawn Burton , Osberger Mary Joe , Segel Phil , Kessler Dorcas
Published: June 2004

Objective: This study compared speech perception benefits in adults implanted with the HiResolutionTM (HiRes) Bionic Ear who used both conventional and HiRes sound processing. A battery of speech tests was used to determine which formats were most appropriate for documenting the wide range of benefit experienced by cochlear-implant users. Study Design: A repeated-measures design was used to assess postimplantation speech perception in adults who received the HiResolution Bionic Ear in a recent clinical trial. Patients were fit first with conventional strategies and assessed after 3 months of use. Patients were then switched to HiRes sound processing and assessed again after 3 months of use. To assess the immediate effect of HiRes sound processing on speech perception performance, consonant recognition testing was performed in a subset of patients after 3 days of HiRes use and compared with their 3-month performance with conventional processing. Setting: Subjects were implanted and evaluated at 19 cochlear implant programs in the USA and Canada affiliated primarily with tertiary medical centers. Patients: Patients were 51 postlinguistically deafened adults. Main Outcome Measures: Speech perception was assessed using CNC monosyllabic words, CID sentences and HINT sentences in quiet and noise. Consonant recognition testing was also administered to a subset of patients (n = 30) using the Iowa Consonant Test presented in quiet and noise. All patients completed a strategy preference questionnaire after 6 months of device use. Results: Consonant identification in quiet and noise improved significantly after only 3 days of HiRes use. The mean improvement from conventional to HiRes processing was significant on all speech perception tests. The largest differences occurred for the HINT sentences in noise. Ninety-six percent of the patients preferred HiRes to conventional sound processing. Ceiling effects occurred for both sentence tests in quiet. Conclusions: Although most patients improved after switching to HiRes sound processing, the greatest differences were seen in the ‘poor’ performers because ‘good’ performers often reached ceiling performance, especially on tests in quiet. Future evaluations of cochlear-implant benefit should make use of more difficult measures, especially for ‘good’ users. Nonetheless, a range of difficulty must remain in test materials to document benefit in the entire population of implant recipients.
Retained heterodisomy for chromosome 12 in atypical lipomatous tumors: implications for ring chromosome formation
Authors: Mertens F. , Panagopoulos I. , Jonson T. , Gisselsson D. , Isaksson M. , Domanski H.A. , Mandahl N.
Published: June 2004

Atypical lipomatous tumor (ALT) is an intermediate malignant mesenchymal tumor that is characterized by supernumerary ring chromosomes and/or giant rod-shaped marker chromosomes (RGMC). Fluorescence in situ hybridization (FISH) and molecular genetic analyses have disclosed that the RGMCs always contain amplified sequences from the long arm of chromosome 12. Typically, RGMCs are the sole clonal changes and so far no deletions or other morphologic aberrations of the two normal-appearing chromosomes 12 that invariably are present have been detected. The mechanisms behind the formation of the RGMCs are unknown, but it could be hypothesized that RGMC formation is preceded by trisomy 12 or, alternatively, that ring formation of one chromosome 12 is followed by duplication of the remaining homolog. The latter scenario would always result in isodisomy for the two normal-appearing chromosomes 12, whereas the former would yield isodisomy in one-third of the cases. In order to investigate these possible mechanisms behind ring formation, we studied polymorphic loci on chromosome 12 in 14 cases of ALT showing one or more supernumerary ring chromosomes and few or no other clonal aberrations at cytogenetic analysis. The molecular genetic analyses showed that the tumor cells always retained both parental copies of chromosome 12, thus refuting the trisomy 12 and duplication hypotheses.
The most common chromosome aberration detected by high-resolution comparative genomic hybridization in vulvar intraepithelial neoplasia is not seen in vulvar squamous cell carcinoma
Authors: Bryndorf T. , Kirchhoff M. , Larsen J. , Andreasson B. , Bjerregaard B. , Westh H. , Rose H. , Lundsteen C.
Published: June 2004

We analyzed genetic changes in condylomas (four cases), vulvar intraepithelial neoplasia I–III (VIN I–III, eleven cases), and primary vulvar squamous cell carcinomas (VSCC, ten cases) by high-resolution comparative genomic hybridization (HR-CGH) and flowcytometry. All samples were also human papilloma virus (HPV)-genotyped. Gain of chromosome 1, the aberration most often seen in VIN III (67%), was not seen in HPV-positive or -negative VSCCs (0%). Both VIN III and VSCC frequently showed gain of 3q (56 and 70%, respectively). The VIN III samples often demonstrated gain of 20q (56%) and 20p (44%), and the VSCC samples gain of 8q (60%), loss of 3p (50%), and 8p (40%). None of the four most frequent changes in the VSCC samples occurred exclusively in the HPV-positive or -negative samples. As expected, we did not find any cytogenetic changes in condylomas and nearly any changes in VIN I–II.
Mosaicism for an ectopic NOR at 8pter and a complex rearrangement of chromosome 8 in a patient with severe psychomotor retardation
Authors: Felbor U. , Knötgen N. , Schams G. , Buwe A. , Steinlein C. , Schmid M.
Published: June 2004

We describe a 3-year-old girl with severe delays in mental and motor skills, a history of generalized seizures, and subtle dysmorphic features. Conventional cytogenetics revealed a mosaic karyotype. A de novo ectopic NOR at the telomeric region of the short arm of one chromosome 8 (8ps) was found in 90% of lymphocyte and in 98% of fibroblast metaphases. A small NOR-bearing marker chromosome and a large derivative chromosome 8 without short arm satellites (der(8)) were present in the remaining cells. FISH with a probe specific for centromeres 14 and 22 labeled both the telomeric region of 8ps and the small marker centromere. Der(8) included an inverted duplication of 8p and a rearranged duplication of 8q but lacked a second centromere. A subtelomeric probe for 8p revealed a cryptic deletion in 8ps and der(8). Thus, the karyotype represents a combination of submicroscopic partial monosomy 8pter and mosaic trisomy 8.
Isolation and characterization of the Xenopus laevis orthologs of the human papillary renal cell carcinoma-associated genes PRCC and MAD2L2 (MAD2B)
Authors: van den Hurk W.H. , Martens G.J.M. , Geurts van Kessel A. , van Groningen J.J.M.
Published: June 2004

Recently we found that the human papillary renal cell carcinoma-associated protein PRCC interacts with the cell cycle control protein Mad2B, and translocates this protein to the nucleus where it exerts its mitotic checkpoint function. Here we have succesfully isolated Xenopus laevis Mad2B and PRCC cDNAs. The full-length xMad2B cDNA encodes a 211 amino acid protein that is highly homologous to human Mad2B, thus pointing to an important function for this protein in higher eukaryotes. The full-length xPRCC cDNA encodes a 544 amino acid protein. Remarkably, this protein contains an amino-terminal region distinct from that in mouse and human, whereas the C-terminal region is highly conserved. Northern blot and RT-PCR analyses revealed a relatively low expression of both xMad2B and xPRCC in most tissues examined. However, an abundant expression was observed in testis and oocyte, indicating a role in meiotic division processes. Coimmunoprecipitation and immunofluorescence analyses revealed that, despite its distinct amino terminus, the xPRCC-protein is still capable of interacting with xMad2B and of shuttling this protein to the nucleus. Therefore, the well-established animal model Xenopus laevis can be used as a powerful system to study in detail the role of xPRCC and xMad2B in the intricate processes of cell cycle control.
Identifying differentially expressed genes in the mammalian retina and the retinal pigment epithelium by suppression subtractive hybridization
Authors: Schulz H.L. , Rahman F.A. , Fadl El Moula F.M. , Stojic J. , Gehrig A. , Weber B.H.F.
Published: June 2004

Retina and retinal pigment epithelium (RPE) cells are of neuroectodermal origin with highly specialized functions in light perception. Identification and characterization of genes differentially expressed in these cells will greatly aid our understanding of their functional roles in retinal biology. As a source enriched for gene transcripts from the retina/RPE, we generated a human retina and a bovine RPE cDNA library applying the PCR-based technique of suppression subtractive hybridization (SSH). Sequencing of 1,080 retina and 2,350 RPE SSH clones resulted in the identification of 321 and 343 non-redundant human transcripts, respectively. Of these, only 27 genes were in common between the two cDNA libraries. One transcript expressed exclusively in retina and RPE is the novel gene C4orf11 which is comprised of four exons on chromosome 4q21.2. We report the full-length cloning of two isoforms of C4orf11, 919 bp and 857 bp in length, both of which contain four identical open reading frames (ORFs). While ORFs 1 to 3 show no homologies to known proteins or protein domains, ORF4 reveals 50% sequence identity to RPE-spondin, a hypothetical protein on 8q13.3 with unknown function. We demonstrate that both the retina and the RPE SSH cDNA libraries are excellent resources for identifying known and novel genes exclusively or abundantly expressed in the retina/RPE complex. In combination with other approaches such as microarray analysis or serial analysis of gene expression (SAGE), the availability of highly sensitive and specific SSH cDNA libraries will facilitate the comprehensive description of the retina/RPE transcriptome.
Gene mapping of 5S rDNA sites in eight fish species from the Paraíba do Sul river basin, Brazil
Authors: Kavalco K.F. , Pazza R. , Bertollo L.A.C. , Moreira-Filho O.
Published: June 2004

The use of improved cytogenetic techniques such as fluorescent in situ hybridization (FISH) has offered important methodologies for cytotaxonomic and evolutionary studies. In particular, the mapping of 5S rDNA sites has proved to be an excellent marker in the study of different organisms and, more recently, in fish. In the present work, the FISH technique was used to map the 5S rDNA sites in the chromosomes of eight neotropical fish species from the Paraíba do Sul river basin, four of these belonging to the order Characiformes, family Characidae, genus Astyanax (A. scabripinnis, A. parahybae, A. giton and A. intermedius) and four to the order Siluriformes, family Loricariidae (Neoplecostomus microps, Harttia loricariformis, Hypostomus affinis and Upsilodus sp.). Karyotype evolution aspects of the analyzed groups are discussed.
‘Quick Course’ Neoadjuvant Chemotherapy with Cisplatin, Bleomycin and Vincristine in Advanced Cervical Cancer
Authors: Singh Kishor C. , Agarwal Alka , Agarwal Sarla , Rajaram Shalini , Goel Neerja , Agarwal Neera
Published: June 2004

To evaluate the response and safety of ‘quick course’ neoadjuvant chemotherapy, 30 patients with advanced squamous cell carcinoma of cervix were given cisplatin, bleomycin, and vincristine weekly for 3 courses. The response was evaluated by subjective parameters and by standard response criteria. In addition to the marked improvement in symptoms, the overall objective response was 60% with a complete pathological response of 6.6%. Tumor volume decreased significantly (p = 0.002) after chemotherapy. Patients with stage IB and 27% (3 of 11) of patients with stage II disease who became technically stage IB (stage reduction) after chemotherapy underwent surgery. Radiotherapy was given to the remaining patients. All patients tolerated the chemotherapy.
Reprocessed (High-Flux) Polyflux® Dialyzers Resist Trans-Membrane Endotoxin Passage and Attenuate Inflammatory Markers
Authors: Teehan Geoffrey S. , Guo Daqing , Perianayagam Mary C. , Balakrishnan Vaidyanathapuram S. , Pereira Brian J.G. , Jaber Bertrand L.
Published: June 2004

Background: Bacterial contamination of dialysis water can contribute to the chronic microinflammatory state observed in dialysis patients. This study characterized the selective permeability of new and peroxyacetic acid/acetic acid/hydrogen peroxide (Renalin®) reprocessed high-flux, polyarylethersulfone-polyvinylpyrrolidone (Polyflux®-17R) dialyzers after exposure to endotoxin-contaminated dialysate during in vitro dialysis. Clinical correlation with pre-dialysis levels of systemic markers of inflammation, and clearance of middle molecules was also assessed in vivo. Methods: Six hemodialysis (HD) patients were enrolled in the study. After reuses 0, 1, 5, 10, and 15, the dialyzers were reclaimed and submitted to an in vitro dialysis circuit using standard dialysate and blood from healthy volunteers. New and reprocessed dialyzers were sequentially exposed to escalating doses of Pseudomonas aeruginosa endotoxin in the dialysate compartment, and whole blood tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) production was used as an index of reverse passage of endotoxin. In vivo, IL-6, C-reactive protein (CRP) and serum amyloid A (SAA) levels were measured to assess the impact of reprocessing on the systemic inflammatory response. Finally, pre- and post-dialysis samples were collected to measure urea and β2-microglobulin (β2-M) clearances. Results: During in vitro dialysis, blood-side endotoxin levels were undetectable following dialysate contamination. TNF-α production remained unchanged (p = NS), and IL-6 production fell significantly on reuses 0, 1, 10, and 15 (p = 0.03) suggesting membrane adsorption, as a result of reuse-dependent surface binding. In vivo, whereas IL-6 and SAA levels did not significantly differ (p = 0.90 and 0.59, respectively), CRP levels fell near significantly, over the course of 15 reuses (p = 0.06). In vivo, β2-M clearance was not affected by the reuse technique (p = 0.28). Conclusions: This study provides in vitro and in vivo evidence arguing that high-flux Polyflux® dialyzers provide more than adequate dialysis, while preventing the in vitro back-diffusion of bacterial endotoxin despite 15 reuses with Renalin®. Clinically, this may translate into an attenuation of the microinflammatory milieu.
Adenovirus-Encoded Hammerhead Ribozyme to PDGF A-Chain mRNA Inhibits Neointima Formation after Arterial Injury
Authors: Lin Zhi-Hong , Fukuda Noboru , Suzuki Ryo , Takagi Hiroto , Ikeda Yukihiro , Saito Satoshi , Matsumoto Koichi , Kanmatsuse Katsuo , Mugishima Hideo
Published: June 2004

To develop a strategy for gene therapy of restenosis following coronary angioplasty, we examined the effects of a recombinant adenovirus vector encoding a hammerhead ribozyme specific for rat platelet-derived growth factor (PDGF) A-chain mRNA (Ad.Ribozyme) and a control recombinant adenovirus vector encoding the Escherichia coli LacZ gene (Ad.LacZ) on neointima formation in rat carotid artery after balloon injury. Ad.Ribozyme (108 PFU/ml) markedly reduced the increased expression of PDGF A-chain mRNA and protein. Ad.Ribozyme significantly decreased the intima/media ratio (68%) of the injured artery, whereas Ad.LacZ had no effect on the intima/media ratio. Most carotid arteries developed thrombi by 14 days after balloon injury, whereas Ad.Ribozyme completely inhibited thrombus formation. Expression of thromboxane A2 (TXA2) receptor mRNA was significantly increased after balloon injury. Ad.Ribozyme significantly decreased the levels of TXA2 receptor. Expression of prostaglandin I2 (PGI2) synthase mRNA was significantly decreased after balloon injury. Ad.Ribozyme significantly increased levels of PGI2 synthase mRNA after balloon injury. The observation that adenovirus-encoded ribozyme to PDGF A-chain inhibits neointima formation may serve as a novel strategy to prevent restenosis after coronary angioplasty. Inhibition of growth factors by genetic approaches may yield new insights into the mechanisms underlying responses to vascular injury and lead to new therapeutic applications.
Body Mass Index but Not a Polymorphism of the Interleukin-1 Receptor Antagonist (IL-1 RA) Gene Is Associated with Age at Natural Menopause
Authors: Riener Eva-Katrin , Keck Christoph , Worda Christof , Hefler Lukas A. , Tempfer Clemens B.
Published: June 2004

Background: A genetic component of the onset of menopause has been described and several candidate genes have been identified. We hypothesized that carriage of a polymorphism of the interleukin-1 receptor antagonist gene (IL-1 RA) is associated with an early age at menopause. Methods: In a prospective cohort study, 90 consecutive postmenopausal Caucasian women were genotyped by PCR for the presence of an 86-base pair tandem repeat polymorphism in intron 2 of IL-1 RA. Results: We found that 36/90 (40%) women were homozygous for the wild-type allele 1 and 49/90 (54%) women were heterozygous for any of the variant alleles (1/2 [n = 44], 1/3 [n = 3], 2/3 [n = 2]). Two women (2%) were homozygous carriers of the variant allele 2. The wild-type allele 1 was identified on 119 of 180 chromosomes for an allele frequency of 0.66. The polymorphic alleles 2 and 3 were present on 56 and 5 chromosomes, respectively (allele frequencies 0.31 and 0.03, respectively). No correlation between the IL-1 RA genotype and the age at menarche and menopause, the length of the reproductive period, and the number of deliveries and miscarriages was ascertained. As to allele frequencies, homozygous and heterozygous carriers of the variant allele 2 had a median age at menopause of 50 (range 40–48) years, compared to 49.5 (range 39–56) years for women with no allele 2 (p value 0.41). Women with at least one allele 2 had a median age at menarche of 13 (range 10–16) years, compared to 13 (range 10–17) years for women with no allele 2 (p value 0.1). Decreasing body mass index, but not smoking, was correlated with an increasing age at natural menopause (r = –0.23, p = 0.04). Conclusions: Our preliminary data suggest that an 86-base pair tandem repeat polymorphism in intron 2 of IL-1 RA does not modulate the onset and cessation of menses in this cohort of Caucasian women.
Safety and Tolerability of High-Dose Angiotensin Receptor Blocker Therapy in Patients with Chronic Kidney Disease: A Pilot Study
Authors: Weinberg Adam J. , Zappe Dion H. , Ashton Michael , Weinberg Marc S.
Published: June 2004

Background: The progression of renal disease is ameliorated by drugs that inhibit the renin-angiotensin system (RAS). The doses used to slow the progression of renal disease may not completely suppress the RAS for 24 h and may explain why some patients do not obtain optimal renoprotective benefits from therapy. This pilot study was initiated to determine the safety and tolerability of using higher doses, than currently approved by the Food and Drug Administration, for the angiotensin-receptor blocker (ARB) candesartan cilexetil in patients with chronic kidney disease. We hypothesized that higher doses will be safe and well tolerated. Consequently, this should be a viable strategy for larger clinical trials evaluating the preservation of renal function. Methods: Twelve patients (10 males; age = 57 ± 14 years) with a history of diabetic or non-diabetic chronic kidney disease were enrolled in an 8-week open-label trial. Patients received candesartan titrated to a targeted dosage of 160 mg/day (5 times above the currently approved maximum dose) and remained at that dosage for the subsequent 4 weeks. The safety and tolerability of the higher doses were determined by measures of blood pressure, serum creatinine and potassium. Results: Candesartan was well tolerated with no serious drug-related adverse events reported. Serum creatinine concentrations throughout the study were not different (p > 0.05) from baseline levels (2.0 ± 0.5 mg/dl). Plasma potassium concentrations at 160 mg/day candesartan (4.9 ± 0.7 mEq/l) were similar (p > 0.05) to those at baseline (4.8 ± 0.5 mEq/l). Conclusions: The results of this pilot study suggest that supramaximal doses of ARBs are safe and well tolerated in patients with chronic kidney disease, while reducing both blood pressure and proteinuria. This study demonstrates the need to further investigate the optimal dosing strategy for ARBs in reducing the progression of renal disease.
Validity and Reliability of the SF-36 Questionnaire in Patients on the Waiting List for a Kidney Transplant and Transplant Patients
Authors: Gómez-Besteiro M. Inmaculada , Santiago-Pérez M. Isolina , Alonso-Hernández Ángel , Valdés-Cañedo Francisco , Rebollo-Álvarez Pablo
Published: June 2004

Background/Aims: The aim of this study is to explore the validity and reliability of the health-related quality of life SF-36 questionnaire in patients undergoing renal replacement therapy. Methods: A multicenter descriptive transversal study was carried out in Galicia, Spain, with patients undergoing renal replacement therapy. The tool used to measure the health-related quality of life was the authorized Spanish version of the ‘SF-36’ generic health questionnaire. The internal consistency of this survey was determined by means of interscale correlations and Cronbach’s α statistic. Validity was examined with a principal component exploratory factor analysis with Varimax rotation. Results: A total of 213 patients waiting for a kidney transplant and 72 recipients with a functioning renal transplant were studied. All the interscale correlations were positive and significant. The overall statistical value for Cronbach’s α was equal to 0.91 (95% CI: 0.91–0.94) and in all domains this value ranged from 0.7 to 0.92. The factor analysis identified 8 factors that explain 66.6% of the variance, 5 of which consisted of the same structure as 5 factors (domains) of the theoretical model. Conclusion: The SF-36 questionnaire is also a reliable and valid tool when used to measure the quality of life of patients undergoing renal replacement therapy.
Comparison of Pregnancy Outcomes of Progesterone or Progesterone + Estradiol for Luteal Phase Support in ICSI-ET Cycles
Authors: Gorkemli H. , Ak D. , Akyurek C. , Aktan M. , Duman S.
Published: June 2004

Objective: To find out the effect of estradiol with progesterone for luteal phase support in IVF-ICSI cycles. Materials and Methods: Patients were accepted for treatment in the ART unit of Selcuk University, Meram Faculty of Medicine, between January 2001 and March 2003. The study was done in a prospective manner. The age range of 252 women was 19–41 years and the total number of cycles was 310. All patients were treated with a long ovulation induction protocol. Patients were treated and divided into two groups in a randomized manner: group I used only 600 mg/day divided into three equal doses of micronized progesterone vaginally, and group II used transdermal estradiol 100 µg/day + 600 mg/day vaginal micronized progesterone. Results: 310 ICSI cycles were carried out in 252 infertile couples between January 2001 and March 2003. From 22 of these cycles, oocytes were retrieved but no embryos were developed. In the remaining 288 cycles there were embryo transfers. All embryo development was achieved by ICSI treatment. In 148 out of 288 cycles, the luteal phase was supported only by vaginal micronized progesterone (group I). On the other hand, the remaining 140 cycles received vaginal micronized progesterone plus transdermal estradiol 100 µg/day (group II). The number of β-hCG-positive results in group I and group II were 20 (13.5 %) and 54 (38.5%) respectively. Conclusion: Adding estradiol to progesterone for luteal phase support in ICSI-ET cycles may increase implantation and pregnancy rates.
Similar Synapse Density in Layer IV Columns of the Primary Somatosensory Cortex of Transgenic Mice with Different Brain Size: Implications for Mechanisms Underlying the Differential Allocation of Cortical Space
Authors: Gutiérrez-Ospina Gabriel , Uribe-Querol Eileen , Sánchez Norma , Geovannini Helga , Padilla Patricia , Hernández-Echeagaray Elizabeth
Published: June 2004

The relative dimension of the areas constituting the cerebral cortex differs greatly in the brains of different mammalian species. The mechanisms by which such an evolutionary remodeling has occurred is not well understood. To begin exploring possible mechanisms, we took advantage of a transgenic mouse model in which the area of the primary somatosensory cortex (S1) shifts, to some extent independent from the area of the cortex as a whole, as a result of differences in the availability of insulin-like growth factor I (IGF-I). Electron microscopy estimations of synapse density in D3 and C3 cortical columns of the S1 layer IV revealed that this parameter was similar among wild type and transgenic mice with higher and lower availability of IGF-I. Because D3 and C3 columns were larger and smaller than normal in mice with higher and lower IGF-I availability, the total number of synapses contained in the average area of D3 and C3 columns increased and decreased, respectively. No differences in the number and overall arrangement of S1 columns were observed among animal groups. These results suggest that: 1) synapse density is a constant factor within the S1 cortical column structure; 2) the mechanisms and factors regulating cell number and synaptogenesis are affected as columns and cortical areas modify their relative dimensions; 3) altered availability of neurotrophic factors might be associated with changes in areal dimensions; and 4) changes in cortical areal dimensions within single lineages might result from the addition of minicolumns to preexisting columns.
Noradrenaline Decreases Spike Voltage Threshold and Induces Electrographic Sharp Waves in Turtle Medial Cortex in vitro
Authors: Lorenzo Daniel , Velluti Julio C.
Published: June 2004

The noradrenergic modulation of neuronal properties has been described at different levels of the mammalian brain. Although the anatomical characteristics of the noradrenergic system are well known in reptiles, functional data are scarce. In our study the noradrenergic modulation of cortical electrogenesis in the turtle medial cortex was studied in vitro using a combination of field and intracellular recordings. Turtle EEG consists of a low voltage background interspersed by spontaneous large sharp waves (LSWs). Noradrenaline (NA, 5–40 µM) induced (or enhanced) the generation of LSWs in a dose-dependent manner. Pharmacological experiments suggest the participation of α and β receptors in this effect. In medial cortex neurons NA induced a hyperpolarization of the resting potential and a decrease of input resistance. Both effects were observed also after TTX treatment. Noradrenaline increased the response of the cells to depolarizing pulses, resulting in an upward shift of the frequency/current relation. In most cells the excitability change was mediated by a decrease of the spike voltage threshold resulting in the reduction of the amount of depolarization needed to fire the cell (voltage threshold minus resting potential). As opposed to the mechanisms reported in mammalian neurons, no changes in the frequency adaptation or the post-train afterhyperpolarization were observed. The NA effects at the cellular level were not reproduced by noradrenergic agonists. Age- and species-dependent properties in the pharmacology of adrenergic receptors could be involved in this result. Cellular effects of NA in turtle cortex are similar to those described in mammals, although the increase in cellular excitability seems to be mediated by a different mechanism.