bluuebunny/crossref_metadata_2025 huggingface dataset:
Inhibition by colchicine of biliary secretion of diethylmaleate in the rat: Evidence for microtubule-dependent vesicular transport
Authors: Dumont Micheline , D’Hont Scorinnem , Durand-Schneider Anne-Marie , Legrand-Defretin Véronique , Feldmann Gérard , Erlinger Serge
Published: March 2007

It has been proposed that a microtubule-dependent transport of vesicles derived from the Golgi apparatus may play a role in biliary secretion of bile salts and other cholephilic anions. To test this hypothesis, we examined the influence of colchicine and vinblastine, two microtubule inhibitors, on diethylmaleate-induced bile flow and on the biliary secretion of diethylmaleate, an organic anion whose glutathione conjugates may be secreted into bile through the Golgi apparatus and Golgi-derived vesicles. Rats were pretreated with colchicine or vinblastine, and diethylmaleate was injected intraperitoneally at doses of 28 to 400 μmol/100 gm body wt. Basal bile flow was unaffected by colchicine or vinblastine. In contrast, diethylmaleate-induced bile flow and the secretion into bile of diethylmaleate conjugates (estimated by the cation-anion gap in bile) were significantly lower in colchicine-treated and vinblastine-treated animals than in controls. Diethylmaleate-induced bile flow was reduced in proportion to diethylmaleate conjugate secretion. A linear relationship was seen between bile flow and biliary output of diethylmaleate conjugates: this relationship was similar in colchicine-treated or vinblastine-treated animals and in controls. At electron microscopy, diethylmaleate had induced distension of the Golgi saccules of the hepatocytes.

In conclusion, colchicine and vinblastine inhibited the secretion into bile of diethylmaleate conjugates and diethylmaleate-induced bile flow. These results support the view that microtubule-dependent transport of Golgi-derived vesicles is involved in the biliary secretion of diethylmaleate and, perhaps, other cholephilic organic anions. (HEPATOLOGY 1991;14:10-15.)
Clinical significance of two forms of IgM antibody to hepatitis delta virus
Authors: Jardi Rosendo , Buti Maria , Rodriguez-Frias Francisco , Garcia-Lafuente Ana , Sjogren Maria H. , Esteban Rafael , Guardia Jaime
Published: March 2007

Separation of 7-8 S and 19 S forms of serum IgM antibodies to the hepatitis delta virus by rate-zonal centrifugation was carried out on serum from 24 patients with hepatitis delta virus infection: 4 patients with acute, self-limited hepatitis; 5 patients with hepatitis delta virus superinfection progressing to chronicity; and 15 patients with chronic hepatitis delta virus.

The high molecular weight IgM form (19 S) was predominantly detected in acute hepatitis delta virus cases, whereas the low molecular weight (7 S) form was found in chronic hepatitis delta virus cases. The serological profile of these two forms of IgM antibody to hepatitis delta virus was investigated in serial samples from five patients with acute hepatitis delta virus superinfection that evolved to chronic hepatitis delta virus. We found that, in the acute stage of the disease, the 19 S form was predominant, whereas 6 mo later a predominance of 7-8 S IgM was observed. These results suggest that IgM antibody to hepatitis delta virus antibody forms are different in acute and chronic hepatitis delta virus infection and that their detection only helps in differentiating an acute infection from a chronic infection but not a hepatitis delta virus—hepatitis B virus.-HBV coinfection from hepatitis delta virus superinfection in the acute stage of the disease. (HEPATOLOGY 1991;14:25-28.)
Persistence of hepatitis B viral DNA after serological recovery from hepatitis B virus infection
Authors: Blum Hubert E. , Liang Jake T. , Galun Eithan , Wands Jack R.
Published: March 2007

Chronic hepatitis B virus infection is a major medical problem worldwide. Apart from HBsAg carriers, hepatitis B virus has also been identified in some HBsAg—individuals with or without antibodies to viral antigens. The molecular mechanisms underlying hepatitis B virus persistence in HBsAg—individuals are unresolved, however. To identify a possible genetic basis for viral persistence, we cloned the viral genome from the liver of a patient serologically immune to hepatitis B virus infection. DNA sequence analysis of the complete viral genome identified numerous mutations in all viral genes. Analysis of the biological effects of these mutations revealed three major findings: a low level of HBsAg synthesis, absence of HBeAg production and a defect terminating viral replication. These data suggest that mutations accumulating during the natural course of hepatitis B virus infection may be a mechanism underlying viral persistence in HBsAg—individuals, presumably through escape from immune surveillance. (HEPATOLOGY 1991;14:56-62.)
Isolation of a complementary DNA fragment of hepatitis C virus in Taiwan revealed significant sequence variations compared with other isolates
Authors: Chen Pei-Jer , Lin Meei-Hua , Tu Su-Jen , Chen Ding-Shinn
Published: March 2007

To clone and characterize hepatitis C virus strains present in Taiwan, RNA was extracted from liver tissue collected from a patient during the acute phase of posttransfusion non-A, non-B hepatitis. RNA was then subjected to complementary DNA synthesis and the polymerase chain reaction, using primers derived from the original nucleotide sequence of the United States hepatitis C virus strain. A complementary DNA clone, HCV-T3, containing 552 base pairs of hepatitis C virus complementary DNA sequences was isolated and characterized. The homologies in nucleotide sequence between the Taiwan isolate and either the United States or Japan isolate were 80.1% and 91.5%, respectively. However, most of the nucleotide changes occurred in the third base positions, resulting in much higher homologies in amino acid sequence of 91.8% and 97.3%, respectively. Amplification of the less conserved region of hepatitis C virus genome with the polymerase chain reaction was improved by use of primers with nucleotides matched to the local strain. Finally, in addition to the liver and serum, the viral genome was also demonstrated in the spleen tissue by similar methods, suggesting another possible target for hepatitis C viral infection. These findings indicate that there is considerable heterogeneity in hepatitis C virus genomes isolated from different areas of the world. (HEPATOLOGY 1991;14:73-78.)
β-blocker therapy for prophylaxis of initial variceal hemorrhage: Has its time come?
Authors: Reynolds Telfer B.
Published: March 2007

In a double-blind randomized trial, the hemodynamic events following the administration of propranolol (n = 51) or a placebo (n = 51) were prospectively studied in cirrhotic patients with esophageal varices. The hepatic venous pressure gradient, heart rate, and variceal size were determined at the baseline and 3, 12, and 24 months after the beginning of therapy. Baseline values were similar in both groups. At 3 months, the hepatic venous pressure gradient decreased significantly in propranolol-treated patients (from 18.1 ± 4.2 to 15.7 ± 3.4 mm Hg; P < 0.05) but not in patients receiving the placebo (19.6 ± 6.8 to 17.5 ± 5.3 mm Hg; NS). At subsequent time intervals this gradient decreased significantly from the baseline value in both groups. Heart rate decreased significantly in the propranolol-treated group at all times (P < 0.001). Variceal hemorrhage occurred in 13 patients (11 placebo-, 2 propranolol-treated; P < 0.01), all of whom had a hepatic venous pressure gradient > 12 mm Hg. In 21 patients (14 propranolol-, 7 placebo-treated) the hepatic venous pressure gradient decreased to ≤12 mm Hg; none of them bled from esophageal varices, and their mortality rate also decreased. Because most of the bleeding events occurred during the first year (10 placebo-, 1 propranololtreated; P < 0.01), propranolol seems to have its protective effect during the period associated with the largest reduction in the hepatic venous pressure gradient. Because a reduction in the hepatic venous pressure gradient to < 12 mm Hg protects from variceal bleeding and increases the rate of survival, this should be the aim of the pharmacological therapy of portal hypertension.
Interpenetrating polymer networks containing gelatin modified with PEGylated RGD and soluble KGF: Synthesis, characterization, and application in in vivo critical dermal wound
Authors: Waldeck Heather , Chung Amy S. , Kao Weiyuan John
Published: March 2007

The purpose of this study was to evaluate the biocompatibility and the efficacy in wound healing of a gelatin‐based interpenetrating polymer network (IPN) containing poly(ethylene glycol) (PEG)‐ylated RGD and soluble KGF‐1 (RGD‐IPN+KGF). IPNs were applied to full‐thickness wounds on a rat model. Wound healing was assessed through histological grading of the host response and percent area contraction at 2 days, 1 week, 2 weeks, and 3 weeks. A control IPN containing unmodified gelatin (unmod‐IPN) and a conventional clinical bandage were applied to similar wounds and also evaluated. During the first week of healing, the unmod‐IPN and conventional dressing wound showed a greater amount of contraction than that of RGD‐IPN+KGF. However, by 3 weeks the extent of wound contraction was comparable between treatments. The RGD‐IPN+KGF treated wound demonstrated lower macrophage and fibroblast densities at 3 weeks as compared to unmod‐IPN treated wounds. RGD‐IPN+KGF acted as a tissue scaffold while preventing the entry of foreign bodies, advantages not seen with the conventional dressing. The extent of cellularity and extracellular matrix organization was higher for wounds healed with RGD‐IPN+KGF than those healed with unmod‐IPN. These results indicate that both soluble and immobilized bioactive factors can be incorporated into our IPN platform to enhance the rate and the quality of dermal wound healing. © 2007 Wiley Periodicals, Inc. J Biomed Mater Res 2007
Effect of solvent content on resin hybridization in wet dentin bonding
Authors: Wang Yong , Spencer Paulette , Yao Xiaomei , Brenda Bohaty
Published: March 2007

With wet bonding techniques, the channels between the demineralized dentin collagen fibrils are filled with debris, solvent, and water. Commercial adhesives include solvents such as ethanol or acetone to facilitate resin‐infiltration into this wet substrate. Under in vivo conditions, the solvent may be diluted because of repeated exposure of the material to the atmosphere, or concentrated because of separation of the bonding liquids into layers within the bottle. The purpose of this study was to investigate the effect of different concentrations of ethanol (10–50%) on infiltration of the adhesive resin and collagen fibril encapsulation in the adhesive/dentin interface using light microscopy, micro‐Raman spectroscopy, and scanning electron microscopy. The results indicated that under wet bonding conditions the hybridization process was highly sensitive to the initial solvent concentration in the adhesive system. The staining and scanning electron microscopy results showed that the quality of the interfacial hybrid layer was poor at the lower (10%) or higher (50%) ethanol content. Micro‐Raman analysis indicated that there was a distinct difference in the degree of adhesive penetration among adhesives containing different concentrations of ethanol. Adhesives containing 10 or 50% ethanol did not realize effective penetration; the penetration of the adhesive monomers increased dramatically when the initial ethanol content was 30%. The amount of solvents are essential for achieving effective bonding to dentin. © 2007 Wiley Periodicals, Inc. J Biomed Mater Res, 2007
Surface structure and apatite‐forming ability of polyethylene substrates irradiated by oxygen cluster ion beams
Authors: Kawashita M. , Itoh S. , Araki R. , Miyamoto K. , Takaoka G. H.
Published: March 2007

Polyethylene (PE) substrates were irradiated at a dose of 1 × 1015ions/cm2by the simultaneous use of oxygen (O2) cluster and monomer ion beams. The acceleration voltage for the ion beams was varied from 3 to 9 kV. Unirradiated and irradiated PE substrates were soaked for 7 days in a metastable calcium phosphate solution (1.5SBF) that had 1.5 times the ion concentrations of a normal simulated body fluid. The irradiated PE substrates formed apatite on their surfaces, irrespective of the acceleration voltage, whereas unirradiated substrates did not form apatite. This is attributed to the formation of functional groups that are effective for apatite nucleation, such as COOH groups, on the substrate surface by the simultaneous use of O2cluster and monomer ion beams. The apatite‐forming ability of the irradiated PE substrates was improved greatly by a subsequent CaCl2solution treatment. This suggests that Ca2+ions introduced on the substrate surface by the CaCl2solution treatment accelerated the apatite nucleation. It is concluded that apatite‐forming ability can be induced on the surface of PE by the simultaneous use of O2cluster and monomer ion beams. © 2007 Wiley Periodicals, Inc. J Biomed Mater Res, 2007
Diffusion‐weighted MRI (DWI) in the oncology patient: Value of breathhold DWI compared to unenhanced and gadolinium‐enhanced MRI
Authors: Low Russell N. , Gurney Jonathan
Published: March 2007

Purpose

To evaluate the feasibility and added value of single breathhold diffusion‐weighted (DW) imaging (DWI) in oncology patients undergoing abdominal MRI.

Materials and Methods

A total of 169 patients with malignancy underwent abdominal MRI at 1.5T, including T1‐weighted (T1W), T2‐weighted (T2W), and dynamic gadolinium‐enhanced imaging. Axial DWI was performed with a single‐shot spin‐echo (SE) echo‐planar imaging (EPI) sequence using a b‐value of 500 seconds/mm2. A total of 24 slices were obtained during a 20‐second breathhold. Two observers reviewed the conventional MR images for tumor. Next, the DW images were reviewed for additional tumor not depicted on conventional MR images

Results

For the 169 patients, additional tumors were noted on the DW images in 77 (0.46) for observer 1 and 67 (0.40) for observer 2. For observer 1 the additional tumor included lymphadenopathy (47), peritoneal metastases (15), renal (1), liver (12), and osseous (2), while for observer 2 the corresponding values were lymphadenopathy (40), peritoneal (12), renal (1), liver (6), osseous (4), and gastrointestinal (1). The DW images resolved as benign findings noted on the conventional MR images in three patients for observer 1 and four patients for observer 2. The conventional MR exam was entirely normal while the DW images showed tumor in 12 (0.07) patients for observer 1 and 10 (0.06) patients for observer 2.

Conclusion

DWI is feasible in a single breathhold and provides additional clinically important information in oncology patients when added to routine abdominal MR sequences. J. Magn. Reson. Imaging 2007. © 2007 Wiley‐Liss, Inc.
Time‐resolved three‐dimensional contrast‐enhanced magnetic resonance angiography in patients who have undergone a Fontan operation or bidirectional cavopulmonary connection: Initial experience
Authors: Goo Hyun Woo , Yang Dong Hyun , Park In‐Sook , Ko Jae Kon , Kim Young Hwee , Seo Dong‐Man , Yun Tae‐Jin , Park Jeong‐Jun
Published: March 2007

Purpose

To evaluate the usefulness of time‐resolved three‐dimensional (3D) magnetic resonance angiography (MRA) using diluted contrast agent (CA) in patients who had undergone a Fontan operation or bidirectional cavopulmonary connection (BCPC).

Materials and Methods

Time‐resolved 3D MRA (10 dynamic data sets, two seconds per dynamic data set) using parallel imaging and keyhole data sampling was performed on 15 patients (median age = 10 years, range = 1–20 years) who had undergone a Fontan operation (N = 11) or BCPC (N = 4). Diluted gadolinium (Gd) contrast agent (CA) was intravenously injected into the arm and/or leg veins. The flow dynamics and morphology of pulmonary circulation, and lung perfusion were assessed.

Results

Preferential or balanced pulmonary blood flow from each systemic vein was visualized on time‐resolved 3D MRA in all patients. In addition, occlusion/stenosis of the central thoracic vein (N = 4) and pulmonary artery (N = 6), systemic venous (N = 5) and arterial (N = 6) collaterals, and lung perfusion defect (N = 4) were identified. Persistent hepatic venous plexus, pulmonary arteriovenous malformation, and axillary arteriovenous fistula were delineated in three patients, respectively.

Conclusion

Time‐resolved 3D MRA with diluted CA is useful for evaluating patients who have undergone a Fontan operation or BCPC because it can reveal the flow dynamics and morphology of pulmonary circulation, and lung perfusion status. J. Magn. Reson. Imaging 2007. © 2007 Wiley‐Liss, Inc.
Hepatitis B virus C gene heterogeneity in a familial cluster of anti‐HBc negative chronic carriers
Authors: Fiordalisi G. , Primi D. , Tanzi E. , Magni E. , Incarbone C. , Zanetti A. R. , Cariani E.
Published: March 2007

We studied a familial cluster of adult HBV chronic carriers characterized by serological markers of active viral replication, normal or slightly elevated ALT levels and, in four out of five cases, absence of anti‐HBc reactivity. The nucleotide sequence of the pre‐C/C region of HBV was analyzed in three patients, showing the presence of wild‐type HBV sequences accompanied by different deleted molecules. Some of the variant genomes were able to encode proteins retaining HBcAg and/or HBeAg reactivity, whereas others contained deletions leading to the synthesis of truncated proteins that could not be immunoprecipitated by anti‐HBc or anti‐HBe mono‐ and polyclonal antibodies. Defective molecules encoded by the variant C gene sequences might play a role in the determination of the clinical profile observed in the analyzed patients. © 1994 Wiley‐Liss, Inc.
Thirteen current anti‐HIV‐l/HIV‐2 enzyme immunoassays: How accurate are they?
Authors: McAlpine Lindsay , Gandhi Jay , Parry John V. , Mortimer Philip P.
Published: March 2007

The sensitivity and specificity of 13 current anti‐HIV‐1/HIV‐2 screening enzyme immunoassays (EIA) for the detection of anti‐HIV in human serum or plasma were investigated by testing against a panel of 454 well‐characterised serum or plasma specimens. The panel included specimens confirmed to contain anti‐HIV‐1 (n = 96), anti‐HIV‐2 (n = 26), or low levels of anti‐HIV (n = 44) as well as specimens collected during HIV‐1 seroconversion (n = 73). Specimens from unselected blood donors (n = 80) and samples from patients with a range of pathological conditions (n = 135) were also included. Observed sensitivities ranged from 96.9% (Biochrom, UBI, and Vironostika) to 100% (Biotest, Cambridge Biotech, IAF Biochem, Ortho, and Vidas). For the seroconversion specimens, Biotest, Cambridge Biotech, Ortho, and Vidas were most sensitive. Observed specificities ranged from 89.9% (UBI) to 100% (Biochrom and Ortho). One assay (Ortho HIV 1+2 EIA) achieved maximum sensitivity and specificity, and was one of four assays to detect anti‐HIV‐1 early in seroconversion. The accuracy of several of the other EIAs was also quite adequate, so additional factors such as convenience and cost can be considered when choosing an assay from the group evaluated.
Chronic liver disease in Peru: Role of viral hepatitis
Authors: Barhani William B. , Figueroa Rolando , Phillips Irving A. , Hyams Kenneth C.
Published: March 2007

The prevalence of antibodies to hepatitis C virus (anti‐HCV) was determined in 105 patients with biopsy‐proven chronic liver disease and 128 comparison patients without any evidence of liver pathology living in Lima, Peru. Using a second‐generation EIA screening and supplemental immunoblot assay, anti‐HCV was detected in four of 13 patients with chronic hepatitis, in 11% of 85 patients with cirrhosis, and in none of seven patients with hepatocellular carcinoma. Only two (1.6%) comparison patients without liver disease had anti‐HCV. Hepatitis B surface antigen (HBsAg) was found in 23% of patients with chronic hepatitis, 12% of patients with cirrhosis, and three of seven patients with hepatocellular carcinoma. There was no evidence of chronic viral hepatitis or alcohol abuse (reported by one‐third of subjects) in 48% of chronic liver disease patients. These preliminary data suggest that among this South American population neither hepatitis B nor hepatitis C infection is the predominate cause of chronic liver disease and that other infectious or environmental factors may be important. © 1994 Wiley‐Liss, Inc.
Ontogeny of glycoprotein gB‐specific antibody and neutralizing activity during natural cytomegalovirus infection
Authors: Marshall Gary S. , Stout Gordon G. , Knights Mari E. , Rabalais Gerard P. , Ashley Rhoda , Miller Helen , Rossier Edmond
Published: March 2007

The envelope glycoprotein gB (gpUL55) is a candidate for inclusion in subunit cytomegalovirus (CMV) vaccines, although data on gB antibody responses after natural infection are limited. [35S]‐labeled gB was partially purified from cells infected with an adenovirus recombinant expressing gB and used in radioimmunoprecipita‐tion assays to characterize responses in solid organ transplant recipients with primary (n = 11) or secondary (n = 8) CMV infection. Seropositive transplant patients without evidence of infection (n = 5) and healthy seroconverters (n = 7) were also studied. gB antibody developed concurrently with CMV‐specific IgG, IgM, and neutralizing activity in transplant patients with primary infection. Sustained boosts in gB antibody were seen in patients with secondary infection, and healthy seroconverters developed early gB responses. These data imply that gB antibody is an integral part of the humoral response to CMV infection, and, in view of experimental data regarding immunogenicity, support a role for gB in subunit vaccines. © 1994 Wiley‐Liss, Inc.
Seroprevalence of antibodies to human herpesviruses in England and Hong Kong
Authors: Kangro H. O. , Osman H. K. , Lau Y. L. , Heath R. B. , Yeung C. Y. , Ng M. H.
Published: March 2007

The age‐related prevalence of antibodies to herpesviruses was compared in England and Hong Kong. Altogether 327 sera from England and 266 sera from Hong Kong were tested for antibodies to herpes simplex virus (HSV), varicella‐zoster virus (VZV), Epstein‐Barr virus (EBV), cytomegalovirus (CMV), and human herpesvirus 6 (HHV‐6). Herpesvirus infections were common in both countries but generally were acquired earlier and were more prevalent in Hong Kong. Over 90% of children in Hong Kong were infected with VZV, EBV, and HHV‐6 by 8 years of age. HSV and CMV were the least prevalent childhood infections in both countries, although, 30–40% of babies in Hong Kong became infected during the first year of life. CMV infections were rare throughout childhood in the English cohort. Overcrowding and early attendance at kindergarten may aid more efficient and earlier transmission of herpesvirus in Hong Kong. The high prevalence of CMV in particular may have implications for the management of young pregnant women and organ transplant recipients in Hong. © 1994 Wiley‐Liss, Inc.
Varicella‐zoster virus antibody avidity and IgG‐subclass patterns in children with recurrent chickenpox
Authors: Junker Anne K. , Tilley Peter
Published: March 2007

The normal immune response after primary varicella‐zoster virus (VZV) infection includes IgG subclass evolution to predominantly IgG1‐type antibodies, and maturation from low to high avidity antibodies which are maintained for life. Twenty‐three healthy and apparently immuno‐competent children with a history of 2‐5 episodes of chickenpox were studied after repeat disease. Serial sera were tested for VZV‐IgG subclass patterns and VZV IgG and G‐subclass antibody avidity by urea elution enzyme linked immunoassay (ELISA). Of 11 patients studied within 8 weeks of repeat chickenpox (Early Group), mean antibody avidity was significantly lower (31.3 ± 26.81) than control (65.1 ± 12.38) (P < .001). Seven had low avidity antibody (<30 percent) and an abundance of IgG3 which was a pattern like primary chickenpox, and 2/11 had high avidity antibody characteristic of anamnestic responses. Early Group patients and 12 others studied over 8 weeks after repeat disease (Late Group) showed avidity maturation and attrition of IgG subclass antibodies other than IgG1. At least nine children failed to show VZV‐specific secondary (memory) immune responses early in the course of repeat disease. It is possible that failure to maintain or evoke a secondary immune response could explain their susceptibility to repeat chickenpox. © 1994 Wiley‐Liss, Inc.
In vivo antiviral effects of mismatched double‐stranded RNA on duck hepatitis B virus
Authors: Ijichi Katsushi , Mitamura Keiji , Ida Setsuko , Machida Haruhiko , Shimada Kaoru
Published: March 2007

The antiviral activity and ability of mismatched double‐stranded RNA (m‐dsRNA), r(I)n′r(C12‐U)n′ to induce interferon (IFN) were evaluated in ducks chronically infected with duck hepatitis B virus (DHBV). When m‐dsRNA was administered intravenously at a single dose of 5 mg/kg, serum DHBV DNA concentrations decreased significantly for 3 days (P < 0.002). However, the DHBV DNA concentrations returned to the pretreatment levels 4 days after treatment. Inhibition of DHBV DNA replication in the liver was also observed 2 days after treatment. Serum IFN activity peaked 3 hours after administration of m‐dsRNA, then rapidly declined. 2′‐5′ Oligo‐adenylate synthetase (2′‐5′ AS) activity increased gradually after treatment and remained elevated for at least 48 hours. In ducks receiving m‐dsRNA once daily for 7 consecutive days, serum DHBV DNA concentrations on the last day of treatment were decreased by 76 ± 12% (P < 0.05) in ducks that received 0.2 mg of m‐dsRNA per kg and by 65 ± 12% (P < 0.05) in ducks that received 1 mg of m‐dsRNA per kg. This decrease persisted for at least 2 weeks after the cessation of treatment in all ducks. These results suggest that m‐dsRNA effectively inhibits DHBV replication in vivo, and that IFN induction and stimulation of 2′–5′AS activity contribute to the inhibition of DHBV replication by m‐dsRNA. © 1994 Wiley‐Liss, Inc.
Detection of parvovirus B19 in a skin biopsy of a patient with erythema infectiosum
Authors: Schwarz Tino F. , Wiersbitzky Siegfried , Pambor Manfred
Published: March 2007

We report the findings on a skin biopsy taken from a child acutely infected with parvovirus B 19 showing the typical exanthematous rash. By indirect immunofluorescence with a monoclonal antibody to B19, viral capsid proteins were detected in epidermal cells localized mainly in the stratum basale. Additionally, B 19 DNA was detected in epidermal cells of the stratum basale by in situ hydridization using a Dig‐labelled B19 DNA probe. The detection of viral capsid proteins and viral DNA suggests the presence of complete viral particles. It is therefore concluded that B19 plays a direct role in the formation of the exanthematous rash in erythema infection. © 1994 Wiley‐Liss, Inc.
Highly sensitive detection of viral RNA genomes in blood specimens by an optimized reverse transcription‐polymerase chain reaction
Authors: Murakami Shigeki , Takahashi Yoshiyuki , Yoshida Shigeru , Fuke Isao , Ohmae Kozo , Mori Chisato , Takagi Mitsuo , Takamizawa Akihisa , Okayama Hiroto
Published: March 2007

A protocol was developed for a highly sensitive detection of viral RNA in blood specimens by reverse transcription coupled with a nested poly‐merase chain reaction. Using Japanese encephalitis virus (JEV) as a model, the optimized reverse transcription‐polymerase chain reaction (ORTPCR) detects as few as 3–5 virions in 0.1 ml of whole blood specimens. The effectiveness of this assay system is confirmed by diagnosis of human hepatitis C viral (HCV) infection. © 1994 Wiley‐Liss, Inc.
Variability of polymerase chain reaction‐based detection of human papillomavirus DNA is associated with the composition of vaginal microbial flora
Authors: McNicol Patricia , Paraskevas Maria , Guijon Fernando
Published: March 2007

The results of repeated human papillomavirus (HPV) DNA testing were compared to changes in cervical pathology and the composition of vaginal microorganisms. A cohort of 19 women with HPV cervical infections in the absence of cervical intraepithelial neoplasia at enrollment was reexamined on average at 7.3‐month intervals over a 2‐year period. At each follow‐up visit, cytological and colposcopic examinations were done and vaginal microorganisms were assessed quantitatively by Gram staining of secretions, and anaerobic and aerobic culture. HPV genotypes 6, 11, 16, and 18 were detected by polymerase chain reaction analysis using DNA isolated from exfoliated cervical cells. The detection of HPV DNA was significantly associated with carriage of Grade II flora (P < 0.001), isolation of Gardner ella vagina/is (P = 0.03), Ureaplasma urealyti cum (P = 0.04), Candida albicans (P = 0.01), Bac teroides species (P = 0.01), and overgrowth by anaerobes (P = 0.004). Normal vaginal flora, characterised by the predominance of Lactoba cillus species, was significantly associated (P < 0.001) with a negative HPV test. The detection of HPV DNA is associated with the composition of microorganisms present in the vagina at the time of testing. © 1994 Wiley‐Liss, Inc.
Measurement of antibody avidity for hepatitis C virus distinguishes primary antibody responses from passively acquired antibody
Authors: Ward K. N. , Dhaliwal W. , Ashworth K. L. , Clutterbuck E. J. , Teo C. G.
Published: March 2007

A new IgG antibody avidity test for hepatitis C virus (HCV) has been developed and was validated using sera from 12 renal dialysis patients infected with HCV. In primary HCV infection low avidity antibody (mean avidity index 24%) was detected within 50 days of seroconversion whereas in long‐term infection (at least 300 days after seroconversion), the mean avidity index was high (88%); in five patients, the avidity index was shown to increase rapidly as time elapsed after primary infection, whereas immunosuppressive therapy was found to delay maturation of the immune reponse in two further patients.

The assay was then employed to confirm that a spurious outbreak of primary HCV infection in eight bone marrow transplant patients was explicable by passive acquisition of high avidity anti‐HCV after intravenous immunoglobulin therapy. It is concluded that this avidity test will have an important role in the investigation of HCV infection in patients. © 1994 Wiley‐Liss, Inc.
Serological response of paediatric oncology patients to human herpesvirus‐6
Authors: Hermione Lyall E. G.
Published: March 2007

The serological response of paediatric oncology patients to human herpesvirus‐6 (HHV‐6) was investigated at presentation and during treatment. Sera from 66 patients presenting with malignancy and 66 controls were examined for anti‐HHV‐6 IgG by indirect immunofluorescence test (IFA) and enzyme linked immunosorbent assay (ELISA), and for anti‐HHV‐6 IgM by IFA. Serial samples from 45 children on chemotherapy were examined for anti‐HHV‐6 IgG by ELISA and sera from selected patients on chemotherapy were examined for IgM by IFA. The response of these patients to four other herpesviruses was also investigated.

Ninety percent of presenting patients and controls were IgG positive for HHV‐6 by IFA and ELISA. Anti‐HHV‐6 IgG as measured by the HHV‐6 ELISA index declined overtime in patients on chemotherapy. Two presenting controls and one leukaemic child with a primary cytomegalo‐virus seroconversion were anti‐HHV‐6 IgM positive. In the patient group seropositivity to herpesviruses (types 1–6) increased with age, the mean age of those with IgG to HHV‐6 alone was 3.7 years compared to 6.8 years for those with antibodies to all five viruses.

At the time of presentation paediatric oncology patients have a similar serological response to HHV‐6 as age‐matched controls and this IgG response wanes with treatment. Whether this is significant in terms of viral pathogenicity is not known and will require investigation of viral activity in these patients. © 1994 Wiley‐Liss, Inc.
Hepatitis C virus‐polymerase chain reaction of routinely processed liver biopsies
Authors: El‐Batanony Mohamed H. , Savage Kay , Jacobs Ruth , El‐Refaie Amany O. , Squadrito Giovanni G. , Brown David , Saleh Saleh M. , Raouf Ahmad A. , Amer Kawther M. , Dusheiko Geoffrey M. , Scheuer Peter J. , Dhillon Amar Paul
Published: March 2007

The aim of the study was to evaluate the specificity and sensitivity of detection of hepatitis C virus (HCV)‐RNA in formalin‐fixed paraffin‐embedded (FFPE) liver biopsies by polymerase chain reaction (PCR). Routinely processed FFPE diagnostic needle liver biopsies as well as stored serum samples from 43 patients with liver disease were tested for HCV‐RNA by reverse transcription‐nested PCR using the same sets of primers and following strict anticontamination measures.

Twenty‐nine cases were positive and 14 were negative for serum HCV‐RNA. Tissue HCV‐RNA was detected in 17 out of the 29 serum HCV‐RNA‐positive cases but not in any of the 14 serum HCV‐RNA‐negative cases. Compared to se‐rum‐PCR, tissue‐PCR was 100% specific, 58.6% sensitive, and 72% efficient. HCV‐RNA was detected more frequently in biopsies stored for less than 1 year, than in those stored for more than 1 year (P= 0.046). In biopsies stored for up to 1 year detection of HCV‐RNA by PCR was 81.8% sensitive and 90.9% efficient. Short (<0.5 cm) liver biopsies were as sufficient for nucleic acid extraction and amplification as long (>0.5 cm) ones.

It is concluded that following strict anticontamination measures, HCV‐RNA detection by PCR in routinely fixed, processed, and stored diagnostic liver biopsies provides a valuable adjunct to diagnosis of HCV infection. In this study, this option was free from contamination problems, even though routine batch histological processing schedules were used. © 1994 Wiley‐Liss, Inc.
Different antibody response to a neutralizing epitope of human cytomegalovirus glycoprotein B among seropositive individuals
Authors: Ayata Minoru , Sugano Tohru , Murayama Tsugiya , Sakamuro Daitoku , Takegami Tsutomu , Matsumoto Yoh‐Ichi , Furukawa Toru
Published: March 2007

The amino‐terminal portion of human cytomeg‐alovirus glycoprotein B (HCMV‐gB) was expressed as a fusion protein to analyze the neutralizing epitope recognized by human monoclonal antibody C23 and the humoral immune response to this epitope. The linear neutralizing epitope was further localized to the pep‐tide within 17 amino acids (position 68‐84) which were conserved between two HCMV laboratory strains. Ten out of 17 HCMV‐seropositive human sera contained the antibody against this epitope. Although seven sera were negative for reacting with the fusion protein, the viruses isolated from the same patients retained the epitope. The immunogenicity of the epitope and the possible application of C23 human monoclonal antibody for passive immunization against HCMV infections are discussed. © 1994 Wiley‐Liss, Inc.
Evolution rate of hepatitis delta virus RNA isolated in Taiwan
Authors: Chao You‐Chen , Tang Hung‐Shang , Hsu Chung‐Te
Published: March 2007

The complete RNA sequences of hepatitis delta viruses (HDV) isolated at 3 years apart from a chronic delta hepatitis patient in Taiwan were determined. The sequence analysis showed an overall evolution rate of 3.18 × 10−3 substitutions/nucleotide/year. The evolution rates in different parts of HDV RNA varied. The hypervariable region evolved faster (4.55 × 10−3 substi‐tutions/nucleotide/year) than the hepatitis delta antigen (HDAg)‐coding region (2.60 × 10−3 substitutions/nucleotide/year) and the autocata‐lytic region (1.11 × 10−3 substitutions/nucleotide/ year). These data are compatible with the previous finding that the hypervariable region is more divergent than the HDAg‐coding region and the au‐tocatalytic regions among the HDV isolates from different geographic areas. No substitution was found in the four previously identified conserved domains of HDV RNA, further confirming their functional importance in viral replication. The evolution rate of this HDV RNA is higher than that determined from the partial RNA sequences of two Japanese HDV isolates and similar to that found in a Lebanon isolate. Further, it was found that this HDV RNA retained the same microheterogeneities at 15 nucleotide positions detected in the RNA 3 years earlier. It is concluded that HDV RNA in patients' serum is extremely heterogeneous, and that the nucleotide substitutions in certain nucleotide positions likely have conferred evolutionary advantages for HDV. Viral sequence evolution is a possible mechanism for chronic HDV infection. © 1994 Wiley‐Liss, Inc.
Racial differences in the seroprevalence of hepatitis A virus infection in Natal/KwaZulu, South Africa
Authors: Sathar M. A. , Soni P. N. , Fernandes‐Costa F. J. T. D. , Wittenberg D. F. , Simjee A. E.
Published: March 2007

The age‐ and race‐specific seroprevalence of hepatitis A virus (HAV) infection was determined by radioimmunoassay (RIA) in 786 subjects between the ages of 6 months to 60 years. More than 50% of African children were seropositive by the age of 5 years. In blood donors (17–60 years), 50% (93/187) of Whites, 67% (110/163) of Indians, 85% (117/137) of Coloureds, and 91% (115/127) of Africans were seropositive. There was a significant difference in the seroprevalence of HAV infection between White blood donors and blood donors from the other three racial groups [Coloureds (P < 0.0001), Africans (P < 0.0001), and Indians (P < 0.001)] and between Indians and Coloureds (P < 0.0001) and Indians and Africans (P < 0.0001). There was no significance difference in HAV infection between Coloureds and Africans (P < 0.200). Eighty‐seven per cent (32/37) of rural Africans had previous infection. In the African population HAV infection is acquired in childhood. There are significant racial differences in the seroprevalence of HAV infection. The surveillance of HAV infection may be used as a valuable yardstick to monitor the changing standards of hygiene and socioeconomic conditions of a community in transition in South Africa and to make rational public health decisions regarding a hepatitis A vaccination policy. © 1994 Wiley‐Liss, Inc.
Hepatitis C virus infections in transplant patients: Serological and virological investigations
Authors: Maple P. A. C. , McKee T. , Desselberger U. , Wreghitt T. G.
Published: March 2007

Hepatitis C virus (HCV) is transmitted by organs of HCV antibody‐positive donors to transplant recipients. This study investigated the serological and virological responses of 14 initially HCV antibody‐negative transplant patients who received organs from four HCV antibody‐positive donors (A‐D) (before donor screening for HCV infection was introduced in 1991). Second generation HCV enzyme immunoassay (Abbott HCV EIA) was used to detect anti‐HCV antibody. Re‐combinant immunoblot (RIBA‐2; Chiron Corporation) and Wellcozyme Western blot (Wwb) assays were compared as confirmatory assays of positive EIA results. Reverse transcription (RT) followed by “nested” polymerase chain reaction (PCR) was performed to detect viral RNA. HCV RNA was only found in the sera of donors B and C, however, transplantation of organs from all donors resulted in infection of all recipients.

HCV RNA was found in recipient sera within 30 days after transplant and remained detectable throughout the period of sampling. An anti‐HCV antibody response was found in only 6 (of the 14) recipients and only after 300 days. Much longer periods passed before detection of HCV antibody in six recipients. For detection of HCV infection in transplant recipients it is essential that testing for HCV RNA by RT‐PCR is carried out. © 1994 Wiley‐Liss, Inc.
Sequence conservation of the major outer capsid glycoprotein of human group C rotaviruses
Authors: Grice Alexander S. , Lambden Paul R. , Caul E. Owen , Clarke Ian N.
Published: March 2007

Several outbreaks of Group C rotavirus infection have occurred in the United Kingdom, in one instance infection was associated with the death of a 4‐month‐old infant in the Bristol area. The origin of human group C rotavirus is unknown although there has been some speculation that porcine species may be a possible source of human infection. Direct reverse transcription‐polymerase chain reaction sequencing of VP7 genes from two UK outbreaks (Bristol and Preston) and sequence analysis from a sporadic case of infection from Brazil (Belém) showed that each of these genes was identical in size (1,063 bp) and has revealed a surprising level (97.8–99.8%) of gene sequence conservation. Sequence comparisons with an isolate from Japan imply that the human group C rotaviruses so far characterised originate from a recent common ancestor with a worldwide distribution. © 1994 Wiley‐Liss, Inc.
Application of commercial assays to detect IgG antibodies to hepatitis C virus in urine: A pilot study from autopsy cases
Authors: Zhang Xiang , Constantine Niel T. , Li Ling , Abdel‐Hamid M. , Bansal Jaya , Smialek John E.
Published: March 2007

Hepatitis C virus (HCV), the main cause of non‐A, non‐B hepatitis in the United States and possibly in the world, is believed to be transmitted primarily through parenteral exposure. Many screening and supplemental tests are available to detect antibodies to HCV in serum. The ability to use commercial assays to detect antibodies to HCV in urine was investigated in this study. A total of 229 serum/urine matched samples were collected sequentially from forensic autopsy cases examined at the Office of the Chief Medical Examiner, State of Maryland. Testing was performed using the Ortho, Innogenetics, and Abbott second generation HCV screening tests and the INNO‐LIA HCV Ab supplemental assay. Sample volumes were increased for urine testing. Forty‐six of 229 serum samples were positive by screening and confirmed by supplemental tests. The urine samples produced positive results on 44–45 of the same 46 by screening tests and all 46 positives by the supplemental test. There were no false positive samples using urine when compared with the serum pairs. The one false negative sample using urine was still nonreactive when the urine volume was increased to 200 μl using the screening tests. Generally, five times the serum volume was required for the screening tests to be optimal for urine samples. The urine samples were stored under different conditions prior to testing to determine the influence on antibody stability in urine.
Viral specificity of hepatitis E virus antigens identified by fluorescent antibody assay using recombinant HEV proteins
Authors: Purdy Michael A. , Carson Dorrie , McCaustland Karen A. , Bradley Daniel W. , Beach Michael J. , Krawczynski Kris
Published: March 2007

A fluorescent antibody (FA) assay for hepatitis E virus antigen (HEVAg) in infected liver tissue was used to confirm the presence of virus‐specific antigens in hepatocytes during the course of infection. With the cloning of the HEV genome it is now possible to determine which viral antigens are recognized by this FA assay. Recombinant HEV proteins covering the carboxyl half of HEV open reading frame 2 (ORF2) were used in this study to demonstrate that some of the most immunoreactive virus‐specific antigens detected by FA are contained within this region of ORF2 (nucleotides 6169–7126).
Evaluation of commercially available assays for antibodies to HIV‐1 in serum obtained from South African patients infected with HIV‐1 subtypes B, C, and D
Authors: Engelbrecht Susan , de Jager Gerrit J. , van Rensburg Estrelita J.
Published: March 2007

Between 1984 and 1990, virus was routinely isolated and serum collected from patients diagnosed at hospitals in the Western Cape as suffering from AIDS or AIDS‐related conditions (ARC). From these, 17 virus strains were selected at random for sequencing and molecular characterisation of the env gene. The strains were previously characterised as belonging to HIV‐1 subtypes B, C and D. The purpose of the present study was to evaluate retrospectively the serological diagnosis of HIV‐1 in these 17 South African patients. Thirteen anti‐HIV screening assays, including 7 rapid/simple test devices (RTDs), 4 enzyme‐linked immunosorbent assays (ElAs) and 2 Western immunoblot assays were evaluated. Using commercial ElAs, 16 serum samples were HIV antibody‐positive and these results were confirmed by Western immunoblot analysis. Serum from one terminal AIDS patient was found negative with all the serological tests. Some RTDs gave false negative antibody reactions on specimens from patients infected with subtype D strains. To investigate the false negative antibody reactions, the polymerase chain reaction (PCR) was used to amplify, clone and sequence proviral DNA from the immunodominant gp41 region from 7 of the HIV‐1 strains. Two patients, both subtype D strains (D214 and D482) with false negative results in the RTDs, showed a significant amino acid substitution, i.e., substitution of a histidine residue for leucine at env position 607. It was concluded that although there were false negative RTD reactions on patients with HIV‐1 subtype D strains, the commercial ElAs tested are sensitive and are able to detect patients infected with HIV‐1 subtypes B, C and D that are present in South Africa. © 1994 Wiley‐Liss, Inc.
Human rotavirus subgroups and serotypes in children with acute gastroenteritis in Saudi Arabia from 1988 to 1992
Authors: Mohammed Khalid A. , Assouli Sufian M. El , Banjar Zainy M.
Published: March 2007

Rotavirus infection was detected in 524 (42.2%) of the 1,242 stool specimens collected from infants and young children with acute gastroenteritis admitted to a major pediatric hospital in Jeddah, Saudi Arabia, between March 1988 and December 1992. Enzyme‐linked immunosorbent assay (ELISA) and monoclonal antibodies specific for subgroup I and II were used to examine 80 rotavirus positive specimens. Subgroup I was detected in 21 (26.3%) and subgroup II in 49 (61.3%) specimens. Six specimens reacted with both subgroup I and II monoclonal antibodies and four specimens were untypeable. Serotyp‐ing of 355 rotavirus positive specimens using monoclonal antibodies specific for the human rotavirus serotypes 1 to 4 revealed a distribution profile of serotype 1, 53.5%; serotype 2, 6.8%; serotype 3, 5.9%; and serotype 4, 22.8%, along with mixed and untypeable specimens (11%). When the correlation between subgroup and serotype specificities was examined in 62 specimens, all subgroup I specimens were found to be serotype 2 or untypeable and all subgroup II specimens belonged predominantly to serotypes 1 (54.7%) and 4 (9.4%). Serotype 1, followed by, to a lesser extent, serotype 4, exhibited a temporal predominance in the 5‐year investigation. A significant clustering of the various serotypes during the cooler months was evident almost throughout the study, particularly in 1989 and 1990. © 1994 Wiley‐Liss, Inc.
Hepatitis C virus and its genotypes in patients suffering from chronic hepatitis C with or without a cryoglobulinemia‐related syndrome
Authors: Willems M. , Sheng L. , Roskams T. , Ramdani B. , Doutrelepont J. M. , Nevens F. , Durez P. , Treille S. , Adler M. , Desmet V. , Fevery J. , Yap S. H.
Published: March 2007

Recently, evidence has been presented for a possible association between hepatitis C virus (HCV) infection and essential mixed cryoglobulinemia (EMC). Eleven consecutive patients with EMC and two with cryoglobulinemia type I were examined for the presence of markers of HCV infection. Eleven of 13 patients (10 with EMC and 1 with type I cryoglobulinemia) had anti‐HCV antibodies (as determined by a second generation anti‐HCV assay) and HCV‐RNA in plasma or serum. HCV‐RNA was also detected in liver biopsies of five patients.

Genotyping showed that HCV genotype 1 was found in 10 of 11 patients with HCV‐RNA (9 genotype 1b and 1 genotype 1a) and only one patient had HCV genotype 2. However, a similar high prevalence of genotype 1b (100%) was found in a group of 14 consecutive patients with chronic hepatitis C, who had no clinical evidence of cryoglobulinemia. Concomitant infection was present in three patients with genotypes 2, 3 and 4, respectively.

These findings stress the high prevalence of HCV infection in patients with EMC and further study shows that a difference in genotype prevalence was not found between HCV‐related EMC and chronic hepatitis C without clinical manifestations of EMC. © 1994 Wiley‐Liss, Inc.