bluuebunny/crossref_metadata_2025 huggingface dataset:
Integrin‐linked kinase production prevents anoikis in human mesenchymal stem cells
Authors: Benoit Danielle S.W. , Tripodi Margaret C. , Blanchette James O. , Langer Steve J. , Leinwand Leslie A. , Anseth Kristi S.
Published: March 2007

Human mesenchymal stem cells (hMSCs) were infected with an adenovirus expressing integrin‐linked kinase (ILK) to understand the role of cell‐ECM signal transduction cascades in suppressing anoikis. Survivability of ILK‐infected hMSCs encapsulated in poly(ethylene glycol) (PEG) hydrogels, an anoikis‐inducing environment, was sustained at 90% over 7 weeks, and survival was attributed to increased protein kinase B (PKB/Akt) activation. hMSCs encapsulated in RGD‐modified hydrogels induced an upregulation in ILK production, PKB/Akt activation, and subsequent survival to the same extent of ILK‐infected, encapsulated hMSCs. As negative controls, encapsulated hMSCs were infected with cyclization recombinase (a protein not associated with cell survival)‐expressing virus, and uninfected hMSCs exhibited very little ILK production, PKB/Akt activation, and survival (∼55% after 7 weeks). As a measure of cell–matrix interactions, vinculin was also quantified for the encapsulated hMSCs and found to be 30‐fold greater for cells encapsulated in RGD‐modified hydrogels and fivefold greater for ILK‐infected hMSCs than controls, indicating that cell–material interactions are inducing the cell survivability of hMSCs encapsulated in RGD‐modified hydrogels. In sum, ILK infection can support cell survival in the absence of matrix interactions and enable fundamental studies of three‐dimensional cell function in response to extrinsic signals, independently of matrix–ligand interactions. © 2007 Wiley Periodicals, Inc. J Biomed Mater Res, 2007
Disparity between ratios of diameters and blood flows in central pulmonary arteries in postoperative congenital heart disease using MRI
Authors: Ordovás Karen G. , Tan Christopher , Reddy Gautham P. , Weber Oliver M. , Lu Ying , Higgins Charles B.
Published: March 2007

Purpose

To compare the relative severity of stenoses of right or left pulmonary arteries with differences in flow to each lung after repair of congenital heart disease (CHD).

Materials and Methods

A total of 15 patients with postoperative congenital heart disease underwent MRI to evaluate branch pulmonary artery stenoses. Spin‐echo images and MR angiography were used to assess morphology, and velocity‐encoded cine (VEC) MRI was used to measure flow in the right and left pulmonary arteries. The ratios of the narrowest diameters of the right to left pulmonary arteries (R/L size) and right to left pulmonary arterial flow (R/L flow) were compared using Spearman's correlation. F test was used to assess the significance of the regression coefficients.

Results

R/L size ratio varied from 0.50 to 2.66, while the R/L flow ratio varied from 0.36 to 12.02. There was an exponential relationship between R/L size and R/L flow, with r2 = 0.78 and P = 0.001. However, severity of morphologic stenoses was not clinically useful for predicting flow reduction. Prediction residuals ranged from −136% to 54% of the true R/L flow.

Conclusion

Anatomical evaluation of the pulmonary arteries does not predict accurately differential blood flow in patients with pulmonary stenoses. Therefore, blood flow measurements are essential when considering the need for further surgical or interventional procedures. J. Magn. Reson. Imaging 2007. © 2007 Wiley‐Liss, Inc.
What causes diminished corticomedullary differentiation in renal insufficiency?
Authors: Lee Vivian S. , Kaur Manmeen , Bokacheva Louisa , Chen Qun , Rusinek Henry , Thakur Ravi , Moses Daniel , Nazzaro Carol , Kramer Elissa L.
Published: March 2007

Purpose

To investigate whether the loss of corticomedullary differentiation (CMD) on T1‐weighted MR images due to renal insufficiency can be attributed to changes in T1 values of the cortex, medulla, or both.

Materials and Methods

Study subjects included 10 patients (serum creatinine range 0.6–3.0 mg/dL) referred for suspected renovascular disease who underwent 99mTc‐diethylene triamine pentaacetic acid (DTPA) renography to determine single kidney glomerular filtration rate (SKGFR) and same‐day MRI, which included T1 measurements and unenhanced T1‐weighted gradient echo imaging. Corticomedullary differentiation on T1‐weighted images was assessed qualitatively and quantitatively.

Results

SKGFR values ranged from 3.5 to 89.4 mL/minute based on radionuclide studies. T1 relaxation times of the medulla exceeded those of renal cortex by 147.9 ± 176.0 msec (mean ± standard deviation [SD]). Regression analysis showed a negative correlation between cortex T1 and SKGFR (r = −0.5; P = 0.03), whereas there was no significant correlation between medullary T1 and SKGFR. The difference between medullary and cortical T1s correlated significantly with SKGFR (r = 0.58; P < 0.01). In all five kidneys with a corticomedullary contrast‐to‐noise ratio (CNR) <5.0 on T1‐weighted images, SKGFR was less than 20 mL/minute.

Conclusion

In our subject population, loss of CMD with decreasing SKGFR can be attributed primarily to an increased T1 relaxation time of the cortex. Medullary T1 values vary but do not appear to correlate with degree of renal insufficiency. J. Magn. Reson. Imaging 2007. © 2007 Wiley‐Liss, Inc.
Case report: Role of hepatitis E virus in the etiology of community‐acquired non‐A, non‐B hepatitis in greece
Authors: Tassopoulos Nicolaos C. , Krawczynski Krzysztof , Hatzakis Angelos , Katsoulidou Antigoni , Delladetsima Ioanna , Koutelou Maria G. , Trichopoulos Dimitrios
Published: March 2007

The aim of this study was to determine the frequency of hepatitis E virus (HEV) infection in a population of Greek adults with community‐acquired (sporadic) non‐A, non‐B hepatitis found to be seronegative for antibodies to hepatitis C virus (anti‐HCV). All patients admitted to the Liver Unit of Western Attica General Hospital and diagnosed as having acute community‐acquired non‐A, non‐B hepatitis between February, 1986, and May, 1990, were enrolled in follow up studies (n = 66). Nineteen patients with HCV infection and 11 patients with acute non‐A, non‐B, non‐C hepatitis that progressed to chronicity were excluded. Convalescent sera were tested for antibody to HEV (anti‐HEV) by a fluorescent antibody blocking assay in 33 of 36 eligible patients. One of the 33 (3%) patients was found to be positive for anti‐HEV. Anti‐HEV testing of all 20 available serum specimens from this patient showed evidence of anti‐HEV seroconversion at the fourth week after the onset of hepatitis. The patient had not travelled abroad or within Greece or had not had apparent contact with people from foreign countries for the previous 3 months. These data show that HEV infection is not a major cause of community‐acquired non‐A, non‐B hepatitis in Greece. However, the reported case of HEV hepatitis suggests that HEV may retain a low endemicity in Greece. More extensive seroprevalence studies are needed for an accurate estimation of the extent of HEV infection in the southeastern European countries. © 1994 Wiiey‐Liss, Inc.
Etiological role of hepatitis E virus in sporadic fulminant hepatitis
Authors: Nanda Santosh K. , Yalcinkaya Kezban , Panigrahi Aswini K. , Acharya Subrat K. , Jameel Shahid , Panda Subrat K.
Published: March 2007

Non‐A, non‐B hepatitis viruses have been implicated as the etiological agent(s) in up to 60% of patients with fulminant hepatitis. These agents are reported to induce a higher mortality than other causes of fulminant hepatitis. Hepatitis E virus (HEV) and hepatitis C virus (HCV) at present constitute the major identifiable non‐A, non‐B hepatitis agents. Of these, HEV has been established as the sole cause of epidemic hepatitis in Afro‐Asian countries, and fulminant hepatitis has been recorded during such epidemics. However, in sporadic cases, the etiological role of HEV in fulminant hepatitis has remained uncertain. The role of HCV in acute liver disease and fulminant hepatitis remains unclear. The present study was undertaken to investigate the association of HEV and HCV in patients with fulminant hepatitis by direct detection of the viral genome using reverse transcription‐polymerase chain reaction (RT‐PCR). Serum samples from 50 sero‐logically identified non‐A, non‐B fulminant hepatitis cases negative for cryptic hepatitis B virus (HBV) infection examined via PCR were tested for HEV and HCV RNA using RT‐PCR. For HEV primers from the nonstructural region (ORF‐1) were used, and for HCV primers from the highly conserved 5′ untranslated regions were used. The products were analysed using agarose gel electrophoresis and confirmed by hybridisation with radiolabelled internal oligonucleotide probes. HEV was detected in 31 (62%) of the 50 fulminant non‐A, non‐B hepatitis cases. In 18 (36%) cases, HCV RNA was detected. In 11 (22%) of the HCV cases, the HEV genome was also amplified. In 20 (40%) cases, HEV was detected alone. Twelve (24%) patients were negative for all viral hepatitis markers. This study establishes HEV as one of the major agents associated with sporadic fulminant hepatitis in a geographical region where epidemics of hepatitis caused by HEV are frequent. © 1994 Wiley‐Liss, inc.
Seroprevalence studies using a recombinant norwalk virus protein enzyme immunoassay
Authors: Parker S. P. , Cubitt W. D. , Jiang X. J. , Estes M. K.
Published: March 2007

A recombinant Norwalk virus (NV) protein enzyme immunoassay was used to study the age of acquisition of NV IgG in various populations. In London, England, there was little evidence of infection during the first 2 years of life. However, the prevalence of NV IgG rose steadily throughout the period that children attend school, reaching a peak of 70% in the group aged 11–16 years. High levels of maternal antibody were detected in infants aged <3 months. Comparison of the acquisition of antibodies to three strains of human calicivirus in Japanese children in northern Japan indicated that although the majority had experienced infection with strains Japan and UK1 by the age of 12 years, only 22% possessed antibodies to NV. In Australian aborigines NV infection occurs early in life; by the age of 6 years over 90% of children were seropositive. © 1994 Wiley‐Liss, Inc.
Presence of hepatitis B and C viral genomes in US blood donors as detected by polymerase chain reaction amplification
Authors: Liang T. Jake , Bodenheimer Henry C. , Yankee Ronald , Brown Nancy V. , Chang Kenneth , Huang Jiakang , Wands Jack R.
Published: March 2007

Hepatitis C virus (HCV) represents a major cause of posttransfusion hepatitis worldwide. Post‐transfusion hepatitis associated with hepatitis B virus (HBV) continues to occur. HBsAg‐negative donor sera from the Rhode Island Blood Center between 1987 and 1988 were screened using more sensitive techniques to assess the prevalence of low level HBV infection. Group I consists of 866 healthy blood donors without HBV sero‐logic markers, group II consists of 377 donors with ALT elevations (>45 IU/L), group II consists of 148 donors positive for anti‐HBc, and group IV consists of eight donors positive for both surrogate markers. A sensitive monoclonal immuno‐radiometric assay (M‐IRMA) was employed for detection of HBsAg‐associated epitopes (detection limit of 20 pg/ml) in serum. A subset of sera were analyzed for the presence of HBV DNA using the method of anti‐HBs capture of HBV related virions in serum followed by polymerase chain reaction (PCR) amplification. Using these techniques, 0.8% and 1.7% of donors were positive for HBsAg and HBV DNA respectively in group I. In contrast, 0.9% and 9.5% in group II and 0.7% and 18.1% in group III were positive, respectively. There were eight donors with both ALT elevation and anti‐HBc; and four (50%) of these were positive for HBV DNA. In the group with anti‐HBc, the majority (80%) of donors with HBV DNA had either no or low (signal to noise ratio < 10) anti‐HBs titer. Using anti‐HCV testing and reverse transcription‐PCR for detection of HCV genomes, we detected evidence of HCV infection in nine of the 49 donors with low level HBV DNA.
Hepatitis C virus infection detected by antibody tests and the polymerase chain reaction as a cause of liver dysfunction in renal transplant recipients
Authors: McCruden E. A. B. , Welch S. , Batchelor B. , Allison M. C. , Dow B. , Follett E. A. C. , Mills P. R. , Briggs J. D.
Published: March 2007

Hepatitis C infection (HCV) is more prevalent in patients who have received kidney transplants than in the general population but the morbidity and mortality associated with infection in this group is unclear. Sera taken from 36 renal transplant recipients with chronic liver dysfunction and from 42 with normal liver function were tested for HCV infection by second generation ELISA (Abbott Laboratories) and second generation recombinant immunoblot assay (Chiron Corporation) (RIBA‐2). Evidence of HCV replication was sought by reverse transcription poly‐merase chain reaction (RT PCR) using primers from the 5′ nontranslated region (5′NTR).

Infection was detected in 20/36 (54%) and in 2/42 (4.8%) controls (P < 0.01). Twelve liver dysfunction patients were positive by all three tests, six were positive by ELISA and RT PCR but had indeterminate RIBA‐2, one was positive by ELISA and RIBA but negative by RT PCR, and one was positive only by RT PCR. Of two infected control patients, one was positive by all three tests and one who was later found to have been in the early stage of infection was positive only by RT PCR. Follow‐up of infected patients showed persistence of viraemia in 14/15 (93%). Evidence of infection with different types of HCV was shown by the lack of amplification by RT PCR by primers with mismatching bases with HCV types 2 and 3.

It is concluded that in our renal transplant patients, chronic HCV infection is usually associated with liver dysfunction and persistent infection is common. © 1994 Wiiey‐Liss, Inc.
Conservation of precore and core sequences of hepatitis B virus in chronic viral carriers
Authors: Hawkins A. E. , Gilson R. J. C. , Bickerton E. A. , Tedder R. S. , Weller I. V. D.
Published: March 2007

Mutations in the precore region of hepatitis B virus (HBV) have been associated with failure of expression of HBV e‐antigen (HBeAg), however, the prevalence of these and other mutations in HBV carriers without overt chronic liver disease remains uncertain. Homosexual or bisexual males (n = 65) with chronic HBV infection attending The Middlesex Hospital, London were studied, of whom two had clinical evidence of chronic liver disease. HBV DMA was amplified from 62 of 65 serum samples using nested and double nested polymerase chain reaction (PCR) assays. Direct sequencing of the PCR products was employed to investigate sequence variation. HBV‐DNA from all available HBeAg‐negative (n = 9) and selected HBeAg‐positive (n = 33) sera were sequenced in the entire precore gene, the 3′ terminal portion of the X gene (aa128–154), and the 5′ terminus of the core gene (aa18–73). Sequences were highly conserved in all regions studied. Samples from two anti‐HBe‐seropositive patients contained mutations in the precore region. In one, a single mutation in the first amino acid resulted in a change to leucine, which would prevent translation of this region and therefore HBeAg expression. Wild type sequences were also detected in this sample. In the other sample from a patient with overt chronic liver disease, a mutation of precore amino acid 28 changed a tryptophan residue to a stop codon which would also prevent HBeAg expression. Although few such patients were studied, precore mutations may be uncommon in anti‐HBe‐seropositive patients without overt chronic liver disease. Possibly such mutations are not related to HBeAg to anti‐HBe seroconversion, but rather they may arise in patients who remain anti‐HBe seropositive for prolonged periods and they may be causally associated with the development of chronic liver disease. © 1994 Wiley‐Liss, Inc.
Hepatitis B virus infection in a group of heterosexuals with multiple partners in Amsterdam, The Netherlands: Implications for vaccination?
Authors: Van Doornum G. J. J. , Van Haastrecht H. J. A. , Hooykaas C. , Van den Hoek J. A. R. , Van der Linden M. M. D. , Coutinho R. A.
Published: March 2007

The aim of the study was to assess prevalence and incidence of hepatitis B virus (HBV) infection among heterosexual men and women with multiple partners attending a sexually transmitted disease (STD) clinic and to establish risk factors of HBV infection in order to consider immunisation for those subjects. A prospective study of heterosexual men and women selected on having multiple partners and presenting to an STD clinic as new patients was carried out from October 1987 through December 1989. Follow‐up continued until December 1990 at the STD clinic of the Municipal Health Service of Amsterdam.

Five hundred ninety‐eight men and women entered the study. More than 70% of both women and men had had commercial sexual partners in the last 5 years. Three hundred eighty‐one participants were born in HBV low endemic countries, 205 came from HBV intermediate endemicity regions. The prevalence of HBV markers in both men and women from low endemic regions was 10%, and for men and women from middle endemic regions 42% and 19%, respectively.

Logistic regression analysis showed that number of years involved in commercial sex was an independent risk factor in male participants from HBV low endemic regions (odds ratio [OR] 1.10 per year) and for women sexual contact with men at high risk of HBV infection (OR 2.59). In people from middle endemic regions more men than women had HBV markers, HBV‐positive men were older than HBV‐negatives (OR 1.05 per year), and for HBV‐positive women the number of years involved in commercial sex was an independent predictor (OR 1.23 per year).

No new cases of HBV infection were found in both groups (upper 95% limit of confidence 7.1 per 1,000 and 35.8 per 1,000 for the participants from low and middle endemic countries, respectively). The duration of follow‐up was 419.9 person‐years at risk for the group from low endemic regions and 83.7 person‐years for the people from middle endemic regions. The participants must be considered to have been at continuous high risk of heterosexual transmission.

The incidence of HBV was so tow that we decided for the moment not to offer hepatitis B vaccination to all heterosexual men and women attending our STD clinic. © 1994 Wiley‐Liss, Inc.
Serological evidence of hantavirus disease in Northern Ireland
Authors: McKenna P. , Clement J. , Matthys P. , Coyle P. V. , McCaughey C.
Published: March 2007

Since, to our knowledge, no clinically documented cases of haemorrhagic fever with renal syndrome (HFRS) have been reported in Northern Ireland, a sero‐epidemiological study was carried out to assess the degree of Hantavirus immunity in a group of 627 Northern Irish patients presenting with symptoms suggestive of HFRS and 100 healthy controls.

IFA screening for IgG Hantavirus specific antibodies was carried out with a panel of up to 9 different Hantaviral antigens. IgM screening was performed using a commercially available ji‐capture ELISA based upon two recombinant Hantaviral nucleocapsid antigens. A seropositivity of 2.1% (15/727) was recorded, with an almost exclusive reaction against a rat‐derived R22VP30 strain of the Seoul serotype. Sole reliance upon non‐rat‐derived classic screening antigens Hantaan (HTN 76–118) and Puumala (CG 18–20) would have resulted in the detection of only 2/15 (13.3%) of cases in IgG IFA, and 8/14 (57.1%) of the cases in IgM ELISA.

The findings indicate that for the first time in Europe, and more specifically in N. Ireland, non‐laboratory outbreaks of HFRS may be caused by wild rats acting as a reservoir for a Seoul‐like Hantavirus.

Conventional Hantavirus serology using Hantaan and Puumala as screening antigens does not appear sufficient for the detection of such cases of HFRS. Hence, we propose the addition of a rat derived Hantaviral antigen to the antigen screening panel as a means of improving the specificity of the detection methods. © 1994 Wiley‐Liss, Inc.
Removal/neutralization of hepatitis A virus during manufacture of high purity, solvent/detergent factor VIII concentrate
Authors: Lemon Stanley M. , Murphy Paula C. , Smith Andrew , Zou Jinsheng , Hammon John , Robinson Stephen , Horowitz Bernard
Published: March 2007

Recent reports have suggested an increased risk of type A viral hepatitis in hemophilic patients treated with high purity factor VIII concentrates prepared using ion exchange chromatography coupled with solvent/detergent treatment for in‐activation of viruses. To determine the capacity for removal or inactivation of hepatitis A virus during the factor VIM manufacturing process, human plasma and various factor VIM production intermediates were spiked with cell culture‐propagated virus and subjected to scaled down conditions mimicking the manufacture of solvent/ detergent factor VIII. The combination of antibody‐mediated neutralization, cryoprecipitation, anion exchange chromatography, and lyophilization in the absence of sucrose resulted in a minimal reduction of 5.5 to 8.55 Iog10 in the infectivity of hepatitis A virus. © 1994 Wiley‐Liss, Inc.
A sensitive PCR method for detecting HCV RNA in plasma pools, blood products, and single donations
Authors: Saldanha J. , Minor P.
Published: March 2007

Although current manufacturing processes appear to efficiently inactivate hepatitis C virus (HCV), it is possible that contaminated blood products may result from failure of some stage during manufacture or from virus overload of plasma pools used for preparation of products. While antibody screening probably removes the majority of HCV positive donations, some donations which are antibody‐negative but HCV positive may be included in pools. The HCV RNA content of plasma pools from paid and voluntary donors was investigated by poly‐merase chain reaction (PCR). A sensitive PCR method using a single pair of primers from the 5′ non‐coding region of the HCV genome and a “hot‐start” was established and shown to be as sensitive as the more conventional nested PCR (which uses two pairs of primers). The majority of pools from paid donors (prescreening) were HCV RNA positive, while all pools from voluntary donors were both antibody and RNA negative. Intravenous immunoglobulins prepared from contaminated pools were RNA negative despite having high antibody levels, indicating satisfactory clearance of the virus during manufacture. The virus load of the pools was at least a thousand‐fold lower than that of single donations, possibly as a result of treatment during the production of the pools or the presence of factors in pools which reduce the sensitivity of some part of the PCR assay. The HCV content of a plasma donation was determined as 3.6 × 106 genomes/ml by an end point dilution method. Thus a simple and sensitive PCR assay was established for detecting HCV RNA in plasma pools and blood products. © 1994 Wiley‐Liss, Inc.
Community prevalence of hepatitis C viraemia: A polymerase chain reaction study
Authors: Sallie Richard , King Ruth , Silva Eduardo , Tibbs Christopher , Johnson Phillip , Williams Roger
Published: March 2007

In order to estimate the prevalence of HCV carriage in an inner city health district, we undertook a polymerase chain reaction (PCR) based survey of sera collected from 1,002 patients attending general practitioners for reasons unrelated to liver disease. The series comprised 305 sample selected sera patients sample from sera 995 patients previously screened by C100 antigen‐based anti‐HCV tests. Overall, 7 patients were positive for HCV RNA. Four cases had anti‐C100 antibodies to HCV, 2 were strictly negative but had high‐normal/borderline optical densities by ELISA assay, while one was completely anti‐HCV negative. All but one had normal liver function tests. Only 3/7 PCR positive cases had any serum marker for hepatitis B (HBV) exposure (2 HBsAg positive, 1 IgM anti‐HBc positive). The minimum point prevalence of HCV carriage in this community is 0.7%, approximating the HBsAg carriage in the same population (1%). HCV carriage in this inner city population is considerably higher than would be predicted by blood donor surveys. A positive anti‐HCV antibody (anti‐C100) test is poorly predictive (∼10%) of HCV RNA carriage in a general practice based population in which measurement of “surrogate” (HBV related) HCV markers would have detected only 3/7 cases of presumed chronic HCV carriage. © 1994 Wiley‐Liss, Inc.
High prevalence of HHV‐6 DNA in peripheral blood mononuclear cells of healthy individuals detected by nested‐PCR
Authors: Cuende José I. , Ruiz Juan , Civeira Maria P. , Prieto Jesús
Published: March 2007

The aim of the study was the evaluation of human herpesvirus‐6 (HHV‐6) infection rate and semiquantification of viremia in healthy people. Healthy blood donors were studied. Human herpesvirus‐6 IgG and IgM antibody titers were measured by indirect immunofluorescence assay. Human herpesvirus‐6 DNA amplification (nested‐PCR) was performed in peripheral blood mononuclear cells (PBMC) and in serum. Seventeen of 50 (34 percent) individuals were positive for IgG anti‐HHV‐6 and the titers ranged from 1:40 to 1:160. None of 30 individuals was positive for IgM anti‐HHV‐6, suggesting no recent infection nor reactivation. Human herpesvirus‐6 DNA was detected by nested‐PCR amplification in peripheral blood mononuclear cells but not in sera. When 1 μg DNA was amplified, HHV‐6 DNA was detected in 8 of 20 individual (40%), but in 18 of 20 (90%) when 5 μg DNA were amplified. It is concluded that HHV‐6 is present in a high proportion of the healthy population but in minimal amounts, and although it can be detected in 1.4 × 105 PBMC, 7 × 105 cells are necessary to detect most cases. No reactivation was observed in healthy people. © 1994 Wiley‐Liss, Inc.
Clinical significance of hepatitis C antibodies in blood donors
Authors: Kolho E. , Färkkilä M. , Mäkinen J. , Krusius T. , Naukkarinen R. , Ikkala E.
Published: March 2007

The clinical significance of hepatitis C antibodies (anti‐HCV) in a healthy population was studied by liver function tests and liver biopsies. The patient population consisted of 195 (96.1%) of the 203 blood donors found to be either anti‐HCV positive or indeterminate by a recombinant im‐munoblot assay (RIBA) during the first year of anti‐HCV screening of 307,606 donors in Finland using a first generation enzyme‐linked immunosorbent assay. Alanine aminotransferase (ALT) levels in 67 donors reacting positively and in 128 reacting indeterminately by a second generation RIBA (RIBA‐4) were monitored to evaluate the prevalence of liver damage. Serum N‐terminal type III procollagen (PIIINP) concentrations were measured in all donors who fulfilled our criterion for possible hepatitis C (ALT values over two times the normal upper limit on two occasions or over five times the normal upper limit on one occasion) and in 23 randomly selected RIBA‐4 positive donors without ALT abnormalities (control group). Two (1.6%) of the RIBA‐4 indeterminate donors had ALT values compatible with possible hepatitis C (negative by polymerase chain reaction) whereas there were 25 (37.3%) such individuals among the RIBA‐4 positive donors (P< 0.0005). Twenty (80%) of the latter 25 RIBA‐4 positive donors with possible hepatitis C consented to liver biopsy. Of these 20 donors, 11 (55.0%) were found to have chronic persistent hepatitis, four (20.0%) mild, three (15.0%) moderate, and two (10.0%) severe chronic active hepatitis. Serum PIIINP concentrations did not correlate with histological score even though the mean concentration was significantly higher in RIBA‐4 positive donors with possible hepatitis C than in the control group (P<0.01). We conclude that HCV antibodies found coincidentally in Finnish donors seldom indicates severe liver disease. © 1994 Wiley‐Liss, Inc.
Prevalence of hepatitis‐C virus RNA in serum and throat washings of children with chronic hepatitis
Authors: Puchhammer‐Stöckl Elisabeth , Mor Wolfgang , Kundi Michael , Heinz Franz‐Xaver , Hofmann Hanns , Kunz Christian
Published: March 2007

Serum samples from 46 children with chronic and probably transfusion acquired hepatitis were tested for the presence of hepatitis C virus (HCV) RNA by a “nested” polymerase chain reaction (PCR) assay, to judge a possible risk of HCV transmission from these patients. In 73% of the samples, viral RNA was detected, indicating a high virus prevalence in this patient group. High titers of HCV‐RNA were observed in some sera as shown by the detection of virus in some samples even at dilutions of 10−3. Comparison of simultaneously obtained PCR results and ALT values revealed no significant correlation between virus presence in serum and higher ALT levels. It was, however, shown that unusually high ALT values may reflect a high titer of viral RNA in serum. To investigate the prevalence of viral RNA in saliva, which could be a vehicle of virus transmission, 35 throat washing samples from the HCV‐infected children were screened by PCR. Using three different sample preparation procedures, 20% of the throat washings were found to be positive for HCV‐RNA. This indicates a prevalence of virus in this fluid lower than that reported previously. © 1994 Wiley‐Liss, Inc.
Prevalence of human T‐Cell lymphotropie virus infections in Germany
Authors: Schätzl Hermann , Schwarzfischer Gerhard , Rose Dietlinde , Gathof Birgit , Weise Wilhelm , Deinhardt Friedrich , Von Der Helm Klaus
Published: March 2007

The extent of human T‐cell lymphotropic retorvirus HTLV‐I and HTLV‐II infections in the general population in central Europe has not been investigated fully. Two hundred forty‐eight thousand blood donors from southern Germany were examined serologically for antibodies to the human lymphotropic retroviruses HTLV‐I and HTLV‐II: 0.021% were confirmed postive and 0.056% were “indeterminate”. A limited number of seropositives and “indeterminate” samples were analyzed by polymerase chain reaction (PCR): the seropositives were confirmed as positive and 43% of the “indeterminate” samples were PCR‐positive. The range of 0.021% HTLV‐positives in 248,000 donors, i.e. about two in 10,000 individuals, mirrors closely the published data for the United States. © 1994 Wiley‐Liss, Inc.
Hepatitis B virus subtypes and hepatitis C virus genotypes in patients with chronic liver disease or on maintenance hemodialysis in Indonesia
Authors: Hadiwandowo Soeliadi , Tsuda Fumio , Okamoto Hiroaki , Tokita Hajime , Wang Yu , Tanaka Takeshi , Miyakawa Yuzo , Mayumi Makoto
Published: March 2007

Hepatitis B surface antigen (HBsAg) and hepatitis C virus (HCV) RNA were surveyed in patients in Yogyakarta, Indonesia, and their subtypes and genotypes were determined by serological methods and polymerase chain reaction with type‐specific primers, respectively. Of 149 patients with chronic liver disease including 24with chronic hepatitis, 86 with liver cirrhosis, and 39 with primary hepatocellular carcinoma, HBsAg was detected in 40 (27%) and HCV RNA in 48 (32%); one patient was positive both for HBsAg and HCV RNA. Thus, the cause of chronic liver disease was not identified in 62 (42%) patients. Of 58 patients on maintenance hemodialysis, four (7%) were positive for HBsAg and 44 (76%) for HCV RNA. Subtype adw was found in 34 (74%) of 46 HBsAg samples and adr in five (11%); compound subtypes, such as adyw and adyr were detected in the remaining seven (15%). Among HCV RNA samples from 48 patients with chronic liver disease, 23 (48%) were of genotype II, 17 (35%) of genotype III and one (2%) of genotype V, in a distribution strikingly different from that of 44 samples from patients on maintenance hemodialysis, 39 (89%) of which were of genotype I and only one (2%) of genotype II. Genotypes were not classifiable in seven (15%) patients with liver disease and four (9%) patients on hemodialysis despite high HCV RNA titers in them all. These results indicate that different HCV genotypes prevail in patients with distinct diseases, as well as unclassifiable HCV genotypes in Indonesia. © 1994 Wiley‐Liss, Inc.
Vitreous fluid sampling and viral genome detection for the diagnosis of viral retinitis in patients with AIDS
Authors: Mitchell Suzanne M. , Fox Julie D. , Tedder Richard S. , Gazzard Brian G. , Lightman Susan
Published: March 2007

Cytomegalovirus (CMV) causes severe necrotizing retinitis in patients with the acquired immune deficiency syndrome (AIDS) and other herpesviruses have been implicated in the acute retinal necrosis syndrome (ARN), seen in both the immunocompetent and the immunosuppressed. At present the diagnosis of viral retinitis relies solely on clinical appearances. In order to assess whether the detection of herpesvirus‐specific DNA in cell‐free vitreous biopsy samples could be useful in the early diagnosis of viral retinitis, vitreous fluid samples were taken from 100 patients. Fifty patients had AIDS as defined by the Centers for Disease Control, (MMWR 36 (suppl 1S):1S–15S, 1987) and retinal disease. The remainder were not known to be HIV infected and had no clinical evidence of retinal infection. Each sample was tested for the presence of CMV, herpes simplex virus 1 (HSV‐1), varicella‐zoster virus (VZV), Epstein‐Barr virus (EBV), and human herpesvirus 6 (HHV6), by amplification of viral DNA using a sensitive and specific nested poly‐merase chain reaction (PCR). The presence of detectable CMV or VZV DNA was clearly associated with clinical disease whereas the presence of HSV‐1, EBV, and HHV6 sequences were not. Clinical discrimination between CMV‐ and VZV‐associated retinitis was greatly enhanced when the PCR results were taken into consideration. © 1994 Wiley‐Liss, Inc.
Antibody response to adenovirus Elb‐derived synthetic peptides and serum levels of p53 in patients with gastrointestinal and other malignant lymphomas
Authors: Lähdeaho M.‐L. , Lehtinen T. , Aine R. , Hakala T. , Lehtinen M.
Published: March 2007

Serum levels of p53 and antipeptide antibody levels to adenovirus type 12 (Ad12) E1b protein were measured in a case‐control study of 62 newly diagnosed patients with malignant lym‐phoma. While patients with gastrointestinal lym‐phoma did not differ from their matched controls, p53 positive lymphoma patients had significantly (P < 0.04) increased antipeptide IgG antibody levels to the Ad12 E1b. Concomitant detection of serum p53 and antipeptide antibodies to Ad 12 E1b was associated with an increased risk (OR = 17.0, 95% confidence limits 1.5, 58.5) of malignant lymphoma suggesting synergism between expression of Ad12 Elb and accumulation of p53 in patients with malignant lymphoma. © 1994 Wiley‐Liss, Inc.
Hepatitis C transmission in a hemodialysis unit: Molecular evidence for spread of virus among patients not sharing equipment
Authors: Allander Tobias , Medin Charlotte , Jacobson Stefan H. , Grillner Lena , Persson Mats A. A.
Published: March 2007

Three cases of simultaneous seroconversion to hepatitis C virus (HCV) in a hemodialysis unit initiated the investigation of the viral strains of 14 seropositive patients in the unit by nucleotide sequencing. The results showed that five patients had been infected with the same viral strain, and indicated that two other patients were sharing a second strain. Transmission was not related to blood transfusions and not associated with the dialysis machines, but occurred between patients treated on the same shift. The number of cases was higher than expected from the serological data. Thus, spread of virus may occur at high frequencies in environments where parenteral routes are made accessible, in spite of rigorous preventive measures. This may raise concern that non‐transfusion associated spread of HCV may be present and unnoticed in several hospital settings. © 1994 Wiley‐Liss, Inc.
Rapid detection of virus genome from imported dengue fever and dengue hemorrhagie fever patients by direct polymerase chain reaction
Authors: Morita Kouichi , Maemoto Tatsuo , Honda Shoji , Onishi Kenji , Murata Misako , Tanaka Mariko , Igarashi Akira
Published: March 2007

Serum specimens from patients with imported dengue fever and dengue hemorrhagic fever were directly subjected to reverse transcription and polymerase chain reaction (RT‐PCR) without any RNA purification. The amplified virus genome was detected within 3 hours. The results of PCR corresponded closely to the results of virus isolation using cultured mosquito cells, suggesting that direct RT‐PCR procedure greatly facilitates rapid diagnosis of dengue infection. © 1994 Wiley‐Liss, Inc.
Diagnosis of hepatitis C virus‐associated chronic liver disease in India: Comparison of HCV antibody assay with a polymerase chain reaction for the 5′ noncoding region
Authors: Panigrahi Aswini K. , Nanda Santosh K. , Dixit Rajesh K. , Acharya Subrat K. , Zuckerman Arie J. , Panda Subrat K.
Published: March 2007

The relative value of an anti‐hepatitis C virus (HCV) serological assay and reverse transcriptase‐nested polymerase chain reaction assays (RT‐PCR) were investigated for the constant 5′ putative noncoding region of HCV for the diagnosis of HCV‐associated chronic liver diseases in India. One hundred fifteen patients with biopsy proven chronic active hepatitis and 140 cases of cirrhosis of the liver were investigated for anti HCV antibody using a second generation commercial enzyme‐linked immunosorbent assay (ELISA). A proportion of these patients: 42 with chronic hepatitis and 27 with cirrhosis of the liver were analysed further for HCV RNA in the serum using RT‐nested PCR assay. Thirty‐three (12.9%) of the 255 patients were positive for anti‐HCV antibody and 23 of 69 (33.3%) patients were positive for HCV RNA in serum. Fifteen of the 33 (45.5%) anti‐HCV positive patients had HCV RNA in the serum. Eight of 36 (22.2%) HCV seronegative patients tested were found with HCV RNA. This indicates that the diagnosis of HCV infection is not possible if it is based solely on the available serodiagnostic tests. Inclusion of both assays improved the diagnostic efficiency, 18.8% (13/69) were negative for all virological markers associated with HBV and HCV infection. Since a majority of the chronic liver disease patients (143/255 [56%]) were seronegative for either HBV or HCV infection, it is significant that HCV RNA was detected in 38% (8/21) of a randomly selected group from these patients. The antibody assay and PCR were compared using interclass correlation (kappa statistics). Both these tests were found to be independent of each other. Considering the wide strain variation throughout the world, it is suggested that the current commercial anti‐HCV testing is inadequate for the diagnosis of all HCV infections in all geographical regions.
Detection of adeno‐associated virus DNA in human genital tissue and in material from spontaneous abortion
Authors: Tobiasch Edda , Rabreau Michèle , Geletneky Karsten , Laruë‐Charlus Serge , Severin Fabienne , Becker Nikolaus , Schlehofer Jörg R.
Published: March 2007

The human helper virus‐dependent parvovirus, adeno‐associated virus (AAV) has never been associated with disease in humans [Berns et al. (1987): Advances in Virus Research 32:243–306; Siegl et al. (1985): Intervirology 23:61–73]. However, in pregnant mice, infection with AAV induces early abortion [Botquin et al. (1993): Journal of Cancer Research and Clinical Oncology 119:24]. We investigated whether this common human virus may be found in human genital tissue or in curettage material from spontaneous abortion. Using the polymerase chain reaction (PCR) AAV type 2 DNA was amplified in histological sections of 19 of 30 biopsies of the uterine mucosa. In addition, AAV‐2 DNA was detected in abortion material during the first trimester of pregnancy (12/30 cases were positive) but not in material of abortion from the second or third trimester (9 cases). Whereas in tissues from the uterus AAV DNA was found only by PCR, large amounts of viral DNA were detectable by Southern blot analysis in abortion material. In situ hybridization revealed DNA of AAV to be present in the villous moiety (trophoblast) of the placenta but not in the embryo or decidua. In the same cells, AAV proteins (including the replication‐associated rep proteins) were detected by immunofluorescence analysis. These results suggest (1) that AAV infects the uterine mucosa (possibly persistently) and (2) that it can replicate in trophoblast cells. This might disturb placenta development and may play a role in early miscarriage. To assess an immunological correlate of AAV infection of cervix uteri lesions and miscarriage, an enzyme‐linked immunosorbent assay (ELISA) was developed for detecting serum IgG‐ and IgM‐class antibodies to AAV‐2. Compared to a control population, a significantly higher prevalence of IgM type antibodies to AAV‐2 was found in women with spontaneous abortion and cervical intraepithelial neoplasia. Since about 80% of the population are seropositive for AAV‐2 the high prevalence of IgM‐class antibodies might be associated with reactivation of latent AAV or to reinfections due to a decline of IgG‐mediated immunity. In view of the presence of AAV‐2 DNA and proteins in placenta tissue, serological tests might be useful to assess further the hypothesis of a possible role of AAV infection in spontaneous abortion.
Degradation of p53 only is not sufficient for the growth stimulatory effect of human papillomavirus 16 E6 oncoprotein in human embryonic fibroblasts
Authors: Ishiwatari Haruko , Hayasaka Naoto , Inoue Hirokazu , Yutsudo Masuo , Hakura Akira
Published: March 2007

Certain types of human papillomavirus (HPV), such as types 16 and 18, are thought to be responsible for the development of cervical carcinomas. The E6 and E7 genes of these viruses have transforming activities in various cultured cells and their mRNAs and proteins are expressed in almost all cervical carcinoma cells. Inactivation of the tumor suppressor p53 protein by the E6 gene is believed to be critical for transformation by these oncogenic HPVs. To determine whether degradation of the p53 protein is, in fact, sufficient for cellular transformation by the E6 gene, the E6 gene of HPV16 was introduced into human embryonic fibroblasts (HEF) using recombinant murine retrovirus and examined whether reduction of the p53 protein could substitute for the E6 function. It was found that HEF cells transfected with the E6 gene showed an increased saturation density and degraded the p53 protein. However, when expression of the p53 protein in normal HEF cells was suppressed by the antisense oligonucleotide of the p53 gene, growth stimulation was not observed. These results show that the E6 gene stimulates growth of HEF cells, but that this activity involves some other E6 gene‐mediated functions than degradation of the p53 protein. © 1994 Wiley‐Liss, Inc.
Simple procedure for discrimination and typing of enteric adenoviruses after detection by polymerase chain reaction
Authors: Allard Annika , Kajon Adriana , Wadell Göran
Published: March 2007

A procedure was developed for specific discrimination and typing of the enteric human adenoviruses, Ad40 and Ad41, after polymerase chain reaction (PCR) amplification of a sequence in the hexon gene highly conserved among all 47 sero‐types recognised. By Taq I restriction of the 300 bp amplimers, subgenus F DNA could be discriminated from DNAs of adenoviruses belonging to all other subgenera. Discrimination between Ad40 and Ad41 was subsequently achieved by cleavage with either Cfo I, HinP I, Mae III, Mvn I, and/or Rsa I. Thus, PCR detection of viral DNA combined with restriction analysis of amplified products provides a valuable tool for use in epidemiological studies of diarrhoea of adenoviral aetiology. © 1994 Wiley‐Liss, inc.
Activation of a varicella‐zoster virus‐specific IgA response during acute Epstein‐Barr virus infection
Authors: Karner Wolfgang , Bauer Georg
Published: March 2007

The influence of an acute Epstein‐Barr virus (EBV) infection on the serological parameters of persistent varicella‐zoster virus (VZV) and herpes simplex virus (HSV) was studied. Sera from 161 patients with infectious mononucleosis caused by EBV and 178 age‐matched controls were tested for HSV‐ and VZV‐specific IgA. 98.7 percent of VZV‐IgG‐positive controls were negative for VZV‐IgA, pointing to the stringent control of latent VZV in healthy individuals. During acute EBV infection, 33.8% of VZV‐IgG‐positive infectious mononucleosis patients produced VZV‐specific IgA. This result may be explained either by reactivation of VZV due to transient suppression of cellular immune functions during acute EBV infection or by polyclonal stimulation caused by EBV. Due to the high incidence of HSV‐IgA in healthy HSV‐IgG‐positive individuals, only a marginal effect of acute EBV infection on the appearance of HSV‐specific IgA was found. © 1994 Wiley‐Liss, Inc.
Brain and kidney of progressive multifocal leukoencephalopathy patients contain identical rearrangements of the JC virus promoter/enhancer
Authors: Ault Grace S. , Stoner Gerald L.
Published: March 2007

The kidneys of six progressive multifocal leukoencephalopathy (PML) patients were examined by PCR amplification for the presence of JC virus. Amplification of three different areas of the viral genome from multiple samples of each kidney revealed three that were positive for the virus. The use of a PCR‐based typing assay on all tissue samples, and cloned sequences from the viral coding region from each positive kidney showed that the same viral genome was present in the kidney as in the brain of the patient. Regulatory region clones all had the archetypal promoter/enhancer structure. However, when PCR fragments from the regulatory region were digested with a restriction enzyme which cuts in region D, the region most often deleted in PML‐type promoters, a low level of undigested DNA remained. This DNA refractory to digestion had a rearranged sequence identical to that of the unique rearranged promoter in the brain of each patient. © 1994 Wiley‐Liss, Inc.
Serum hepatitis C virus RNA level as a predictor of subsequent response to interferon‐α therapy in Japanese patients with chronic hepatitis C
Authors: Onto Etsuro , Mizokami Masashi , Nakano Tatsunori , Terashima Hisahiro , Nojiri Osamu , Sakakibara Kenji , Mizuno Makoto , Ogino Masataka , Nakamura Makoto , Matsumoto Yukoh , Miyata Ken‐Ichi , Lau Johnson Y. N.
Published: March 2007

Serum hepatitis C virus (HCV) RNA level has been shown to be a good predictor of subsequent response to interferon‐α (IFN) therapy in US patients in whom genotype 1a/1b are both predominant. To determine whether serum HCV RNA level is a predictor of subsequent response to IFN in Japanese patients or not, appropriately collected pre‐IFN therapy serum samples from 35 Japanese patients with chronic HCV infection were studied. Serum HCV RNA level and HCV genotype were determined and correlated with the subsequent response to IFN. Response to IFN was defined by serum alanine transaminase level: complete and sustained response (n = 15), complete response followed by relapse (n = 10), and no response (n = 10). Patients with complete and sustained response had lower pre‐IFN serum HCV RNA level (median: RT‐PCR+, bDNA‐) compared to the complete response to relapse group (median: 5.25 × 106 genome equivalent/ml [eq/ml], P < 0.001) and the no response group (median: 10.63 × 106 eq/ml, P < 0.001). Seven (46.7%) of the complete and sustained response patients had HCV genotype 2a and three patients had a mixture of genotypes 1b and 2a. In contrast, all 10 patients in the complete response to relapse group had genotype 1b whereas 8 of 10 patients in the non‐response group had genotype 1b and 2 had genotype 2b. The patients with HCV genotype 2a had lower serum HCV RNA level than those with 1b (P = 0.002). When the HCV viremia were controlled by stratifying them into < and > 106 eq/ml, patients with genotype 1 had a similar complete and sustained response rate compared to genotype 2. These data indicated that pre‐IFN serum HCV RNA is also a good predictor of subsequent complete and sustained response in Japanese patients with chronic HCV infection. © 1994 Wiley‐Liss, Inc.