Human papillomavirus E6/E7 mRNA expression in women younger than 30 years of age.

OBJECTIVE: The prevalence of human papillomavirus (HPV) is high in women younger than 30 years of age, most infections being transient. It is not clear, however, to what extent the E6/E7 transcripts are being expressed. This may be of prognostic importance. In this study, we have determined the prevalence of HPV DNA and mRNA in 283 women younger than 30 years of age. METHODS: E6/E7 transcripts from HPV types 16, 18, 31, 33 and 45 were detected using PreTect HPV-Proofer, while the presence of HPV DNA was detected using Gp5+/6+ consensus PCR and type-specific PCR. RESULTS: A total of 92 women (32.5%) were positive by consensus PCR, 59 (20.8%) were positive by type-specific PCR, while 41 (14.5%) were positive by PreTect HPV-Proofer. E6/E7 mRNA expression was detected in 38 (64.4%) of the 59 HPV type-specific DNA positive women. For HPV 16, E6/E7 mRNA expression was observed in 8 (32%) of the 25 DNA positive women. No high-grade lesions were observed in the concomitant cytology. CONCLUSIONS: Among young women having a normal Pap smear, a high HPV prevalence was found. Hence, use of consensus PCR will most probably give a low prognostic value for identifying subsequent severe dysplasia. The five HPV types 16, 18, 31, 33 and 45 accounted for the majority of infections with two out of three having a detectable E6/E7 mRNA expression. Yet, repeated type-specific testing for HPV mRNA may identify young women with a persistent transforming infection being at increased risk for severe dysplasia.

Omega- and Omega+ production in central Pb + Pb collisions at 40 and 158A GeV.

Results are presented on Omega production in central Pb+Pb collisions at 40 and 158A GeV beam energy. For the first time in heavy ion reactions, rapidity distributions and total yields were measured for the sum Omega(-) + Omega(+) at 40A GeV and for Omega(-) and Omega(+) separately at 158A GeV. The yields are strongly underpredicted by the string-hadronic UrQMD model but agree better with predictions from hadron gas models.

System-size dependence of strangeness production in nucleus-nucleus collisions at squareroot[sNN]=17.3 GeV.

Emission of pi+/-, K+/-, phi, and Lambda was measured in near-central C+C and Si+Si collisions at 158 AGeV beam energy. Together with earlier data for p+p, S+S, and Pb+Pb, the system-size dependence of relative strangeness production in nucleus-nucleus collisions is obtained. Its fast rise and the saturation observed at about 60 participating nucleons can be understood as the onset of the formation of coherent systems of increasing size.

Lambda and lambda production in central Pb-Pb collisions at 40, 80, and 158A GeV.

Production of Lambda and Antilambda hyperons was measured in central Pb-Pb collisions at 40, 80, and 158A GeV beam energy on a fixed target. Transverse mass spectra and rapidity distributions are given for all three energies. The Lambda/pi ratio at midrapidity and in full phase space shows a pronounced maximum between the highest BNL Alternating Gradient Synchrotron and 40A GeV CERN Super Proton Synchrotron energies, whereas the Lambda/pi ratio exhibits a monotonic increase.

Human papillomavirus oncogenic expression in the dysplastic portio; an investigation of biopsies from 190 cervical cones.

In this study, we investigated the presence of E6/E7 transcripts of seven common high-risk human papillomavirus (HPV) types in 190 cervical biopsies. The RNA-based real-time nucleic acid sequence-based amplification assay (NASBA) and type-specific PCR, both detecting HPV 16, 18, 31, 33, 45, 52, and 58, as well as consensus PCR, were performed on all 190 biopsies. High accordance between type-specific and consensus PCR confirms that the HPV types included in this study are the most common types present in cervical dysplasia. Furthermore, we see a clear increase in the incidence of HPV, both DNA and RNA, along with the histological severity of dysplasia. HPV RNA was detected in all but two PCR-positive cases, confirming that the virus exerts E6/E7 mRNA expression in cases of high-grade dysplasia. Out of 19 women given a normal or borderline diagnosis at conisation, only four were found HPV positive, which may suggest that unnecessary conisations can possibly be reduced by introducing HPV testing into the preoperative routine assessment.

Evidence for an exotic S= -2, Q= -2 baryon resonance in proton-proton collisions at the CERN SPS.

Results of resonance searches in the Xi(-)pi(-), Xi(-)pi(+), Xi;(+)pi(-), and Xi;(+)pi(+) invariant mass spectra in proton-proton collisions at sqrt[s]=17.2 GeV are presented. Evidence is shown for the existence of a narrow Xi(-)pi(-) baryon resonance with mass of 1.862+/-0.002 GeV/c(2) and width below the detector resolution of about 0.018 GeV/c(2). The significance is estimated to be above 4.2sigma. This state is a candidate for the hypothetical exotic Xi(--)(3/2) baryon with S=-2, I=3 / 2, and a quark content of (dsdsu). At the same mass, a peak is observed in the Xi(-)pi(+) spectrum which is a candidate for the Xi(0)(3/2) member of this isospin quartet with a quark content of (dsus[-]d). The corresponding antibaryon spectra also show enhancements at the same invariant mass.

[Portasystemic shunt--our twenty years experience]. / Portosystemische Shuntanlage--ein Rückblick nach 20 Jahren, Germany.

OBJECTIVE: It was the aim of this study to demonstrate our experiences over twenty years with portasystemic shunt surgery in patients with chronic liver disease and variceal bleeding. PATIENTS AND METHODS: From January 1 st, 1980 to December 31 st, 2000 we performed 90 portasystemic shunt operations (PSO). The patients were divided in two groups. The patients of the first group were operated upon between 1980-1988 (n = 58), patients of the second group (n = 32) between 1988-1998. Both groups did not differ in age, gender and cause of hepatic disease. In the first group the most performed type of shunt was the portacaval shunt, in group II the splenorenal shunt. RESULTS: We observed an improved early and late mortality rate, encephalopathy rate and reduction of recurrent variceal bleeding in the second group: the early mortality rate decreased from 16 to 9 % (p < 0.01), the late mortality rate from 35 to 6 % (p < 0.05), the encephalopathy rate from 43 to 12 % and the variceal rebleeding rate from 10 to 6 %. CONCLUSION: Selective shunts, such as the distal splenorenal shunt are significantly superior to the standard (end-to-side or side-to-side) portasystemic shunt. In countries where acute treatment of recurrent variceal hemorrhage with sclerotherapy is not available in remote areas or in countries where transplantation procedures are in the very beginning and where TIPPS operations are too expensive, portasystemic shunt operations are the only possibility to save the patients life when sclerotherapy fails.

Congenital choledochal cysts in adults--comparative study.

Congenital choledochal cysts are rare anomalies of the biliary tree and their presentation in adults is infrequent. They are more common in Asia. Females are more commonly affected. Surgery remains the treatment of choice. Nine patients were operated for congenital choledochal cysts in the last fifteen years, i.e. from 1988 to 2002. The diagnosis was established by case history, clinical features and laboratory tests. The imaging methods proved to be the most informative among them. Classification of the choledochal cysts was based on modified Todani classification. All patients have undergone cyst excision with Roux-en-Y hepaticojejunostomy. The complications, like recurrent cholangitis or pancreatitis, were avoided.

Open reading frame III of borna disease virus encodes a nonglycosylated matrix protein.

The open reading frame III of Borna disease virus (BDV) codes for a protein with a mass of 16 kDa, named p16 or BDV-M. p16 was described as an N-glycosylated protein in several previous publications and therefore was termed gp18, although the amino acid sequence of p16 does not contain any regular consensus sequence for N glycosylation. We examined glycosylation of p16 and studied its membrane topology using antisera raised against peptides, which comprise the N and the C termini. Neither an N- nor a C-terminal peptide is cleaved from p16 during maturation. Neither deglycosylation of p16 by endoglycosidases nor binding of lectin to p16 was detectable. Introduction of typical N-glycosylation sites at the proposed sites of p16 failed in carbohydrate attachment. Flotation experiments with membranes of BDV-infected cells on density gradients revealed that p16 is not an integral membrane protein, since it can be dissociated from membranes. Our experimental data strongly suggest that p16 is a typical nonglycosylated matrix protein associated at the inner surface of the viral membrane, as is true for homologous proteins of other members of the Mononegavirales order.

Multimerization potential of the cytoplasmic domain of the human cytomegalovirus glycoprotein B.

In the present study the coding sequence of the cytoplasmic tail of the human cytomegalovirus glycoprotein B (gB) was expressed. The secondary structure of the purified recombinant protein was analyzed by circular dichroism, and the quaternary structure was investigated by gel permeation chromatography, and electron microscopy. Our data indicate that the cytoplasmic gB domain contains alpha-helix structures and assembles into tetramers, suggesting that the authentic gB may represent a homotetramer.

Antiphospholipid antibody levels in intravenous immunoglobulin (IVIg) preparations.

The antiphospholipid syndrome (APS) is characterized by various clinical manifestations and by elevated levels of antiphospholipid antibodies. Passive induction of APS by infusion of these antibodies has been demonstrated in animal models. Intravenous immunoglobulin (IVIg) is one of the therapeutic options in APS. In this study, five commercially used preparations of IVIg were tested for the presence of elevated levels of cardiolipin, beta-glycoprotein-I, phosphatidylserine, antinuclear, and double-stranded DNA autoantibodies, as well as for lupus anticoagulant activity. The absence of abnormal elevated levels of any of these autoantibodies in five different IVIg preparations provides additional evidence for the safety of IVIg use in APS.

Flutamide-induced acute hepatitis in advanced prostate cancer patients.

OBJECTIVE: To describe the characteristics of 3 patients hospitalized in the Department of Gastroenterology, University Hospital Center, Rijeka, as the result of acute hepatitis, a rare adverse drug reaction to flutamide. PATIENTS AND RESULTS: All 3 patients with advanced prostate carcinoma were treated with oral flutamide at a dose rate of 250 mg 3 times daily. The patients developed clinical signs (jaundice, anorexia, nausea, dark urine etc.) and laboratory liver function test changes (high aminotransferase and bilirubin level), indicative of acute hepatitis, 20-22 weeks after commencing flutamide treatment. The flutamide therapy was immediately discontinued and this resulted in spontaneous remission (clinical and liver function test results returned to normal) during the next 8 weeks. CONCLUSION: Our data clearly suggest that flutamide causes acute hepatitis and that the monitoring of the patients' liver function tests in order to detect these changes as early as possible, is important.

DNA binding-independent transcriptional activation by the androgen receptor through triggering of coactivators.

Androgens have critical roles in the development and maintenance of the male reproductive system and are important for progression of prostate cancer. The effects of androgens are mediated by the androgen receptor (AR), which is a ligand-modulated transcription factor that belongs to the nuclear receptor superfamily. In the presence of androgens, AR binds to androgen response elements in the vicinity of androgen receptor target genes and activates transcription. In addition, liganded AR can interfere with the activity of other transcription factors, such as activator protein-1 and nuclear factor kappaB, for which DNA binding by AR is not necessary. In this study, we describe a novel ligand-dependent transactivation function for AR that is independent of its DNA binding ability. AR dramatically increased the intrinsic transcriptional activity of the nuclear receptor coactivators glucocorticoid receptor-interacting protein-1 (GRIP1), cAMP response element-binding protein-binding protein, and p300 that are tethered to DNA. This "triggering" phenomenon required both similar and distinctly different regions of AR compared with those needed for ligand-dependent transactivation from androgen-responsive elements. Furthermore, the domains of GRIP1 required for triggering by AR are different from those required when GRIP1 serves as a coactivator for AR at androgen-responsive promoters. These data suggest that triggering may constitute an important part of the mechanism by which AR regulates transcription.

Acute hepatitis induced by alpha-interferon in a patient with chronic hepatitis C.

Hepatic adverse effects occur very rarely with alpha-interferon therapy. A case of acute hepatitis induced by alpha-interferon in a 33-year-old man with chronic hepatitis C is described. The patient developed acute hepatitis with very high aminotransferase activity and jaundice. After discontinuing alpha-interferon therapy, hepatitis resolved rapidly. The immune-mediated mechanism is the most probable cause of this hepatitis.

Chiral hedgehog textures in two-dimensional XY-like ordered domains

The textures associated with a point defect centered in a circular domain of a thin film with XY-like ordering have been analyzed. The family of equilibrium textures, both stable and metastable, can be classified by a new radial topological number in addition to the winding number of the defect. Chiral textures are supported in an achiral system as a result of spontaneously broken chiral symmetry. Among these chiral textures, our theoretical analysis accurately describes two categories of recently discovered "reversing spiral" textures, ones that are energetically stable and metastable.

Mutational analysis of the androgen receptor AF-2 (activation function 2) core domain reveals functional and mechanistic differences of conserved residues compared with other nuclear receptors.

A short C-terminal sequence that forms the core of the activation function-2 (AF-2) domain is conserved in members of the nuclear receptor superfamily and is required for their normal biological function. Despite a high degree of sequence similarity, there are differences in the context and structure of AF-2 in different nuclear receptors. To gain deeper insight into these differences, we carried out an extensive mutational analysis of the AF-2 core in the androgen receptor (AR) and compared the changes in transcriptional activity with similar mutations that have previously been generated in other nuclear receptors. Mutagenesis of Met894 to Asp resulted in substantial decreases in both DNA and ligand binding activities and, consequently, a significant drop in ligand-dependent transcriptional activation. In contrast, substitution of Met894 with Ala did not affect DNA or hormone binding, and the transactivation potential was comparable to that of wild-type AR. Mutagenesis of Glu897 either with Val or Ala significantly impaired ligand-dependent activation that was not due to changes in DNA or ligand binding. There are both similarities and distinct differences between these findings compared with previous mutagenesis studies of the corresponding residues in other nuclear receptors. All mutants efficiently interfered with AP-1 activity, indicating that ligand-dependent activation of transcription and interference with AP-1 activity are separable functions in AR. For the Glu897 substitutions, the decrease in ligand-dependent transactivation could partially be reversed by overexpression of GRIP1 (GR-interacting protein 1) or CBP, putative coactivators for AR. However, there was no correlation between ligand-dependent in vitro or in vivo association with coactivators and the ability of the mutants to support ligand-dependent transactivation. This is in contrast to similar mutations in other nuclear receptors that lose interactions with putative coactivators concomitant with their loss of transcriptional activity. However, the Glu897 mutations disrupted the intramolecular interaction between the N-terminal domain and the ligand-binding domain of AR that was recently suggested to be required for normal AR function. We conclude that residues in the AF-2 core domain of AR make distinctly different contributions to its transcriptional activities compared with those of other nuclear receptors studied to date.

Anosmia induced with alpha interferon in a patient with chronic hepatitis C.

This is a report of the alpha interferon-induced acute anosmia in a 37-year old patient with chronic hepatitis C. This exceptionally rare side-effect started in our patient as a smelling problem 2 weeks after the initiation of the therapy, and anosmia is still present 13 months after the discontinuation of the alpha interferon. We presume that neurotoxic mechanism could be responsible for this side-effect.

The N-terminal domain of thyroid hormone receptor-alpha is required for its biological activities.

Thyroid hormone (T3) receptors (T3Rs) are ligand-modulated transcription factors that belong to the nuclear receptor superfamily. Whereas the well-conserved DNA-binding domain and the relatively well-conserved ligand-binding domain in T3Rs have been characterized in detail, limited information is available on the contribution of the variable N terminus to the transcriptional properties of T3Rs. To gain greater insight into the function of the N terminus, we generated a deletion mutant of T3Ralpha, T3Ralpha-deltaN1, that lacks amino acids 7-45 and assessed the effect of this deletion on all known transcriptional activities of T3Ralpha. Despite the fact that T3Ralpha-deltaN1 was expressed and bound T3 with an affinity similar to that of wildtype T3Ralpha, all of its common transcriptional activities were lost. That is, T3Ralpha-deltaN1 did not activate transcription from a positive or negative T3 response element, and it could not interfere with AP-1 transcriptional activity. Surprisingly, T3Ralpha-deltaN1 lost its ability to bind DNA, which can account for its deficiencies as a transcriptional activator. In contrast, the ability of T3Ralpha-deltaN1 to interact with putative coactivators or corepressors was not significantly altered from that of wildtype T3Ralpha. However, overall folding of T3Ralpha-deltaN1 was altered, as indicated by differential sensitivity to limited protease digestion. These data document that the N terminus of T3Ralpha, albeit relatively short and representing a variable and unconserved region when compared with other nuclear receptors, has a critical role in proper folding of the DNA-binding domain and is required for the biological activities of full-length T3Ralpha.

Family study of girls with attention deficit hyperactivity disorder.

OBJECTIVE: Because attention deficit hyperactivity disorder (ADHD) is relatively infrequent among girls, little is known about the causes of ADHD in girls. To help fill this gap in the literature, the authors assessed the familial transmission of ADHD in families ascertained through girls. METHOD: Interviewers who were blind to diagnosis administered structured psychiatric interviews to 140 girls with ADHD and their 417 first-degree relatives and to 122 girls without ADHD and their 369 first-degree relatives. RESULTS: The relatives of the ADHD girls had a significantly higher prevalence of ADHD, according to either the DSM-III-R or DSM-IV definition, than the relatives of the comparison girls. However, this did not differ from the prevalence the authors reported previously for families of boys with ADHD. Like the boys' families, the relatives of the girl probands also had significantly higher prevalences of antisocial, mood, anxiety, and substance use disorders, although the prevalence of familial antisocial disorders was lower than had been observed in the boys' families. There was no association between the DSM-IV subtypes of the probands and relatives. CONCLUSIONS: The familial transmission of ADHD and comorbid disorders generalizes to families of girls with ADHD. Neither proband gender nor subtype influences the familial transmission of ADHD.

Mutational analysis of two PWW sequence motifs in human aminoacylase 1.

Human and porcine aminoacylase 1 (Acy1) contain two peculiar sequence motifs located near the interface between the two major domains of the Acy1 subunit. Each motif consists of the sequence PWW preceded by four strongly polar residues. In order to examine the significance of these sequences for Acy1 stability and/or catalysis, we used site-directed mutagenesis of human Acy1 to replace the tryptophan residues in either motif with alanines. Both mutants showed unchanged zinc binding and normal substrate affinity. Modification in PWW motif 1 (residues 192 -194) and motif 2 (residues 321 - 323) resulted in markedly reduced specific activity in the first case and diminished stability in either mutant. Fluorescence quenching measurements showed that all four tryptophans of the PWW motifs are solvent-accessible. We conclude that PWW motif 1 maintains the native conformation of the active site by creating the proper spatial relationship between dimerization domains and catalytic domains, while motif PWW2 is necessary for the stability of the catalytic domain.