bluuebunny/crossref_metadata_2025 huggingface dataset:
Sonography versus helical CT in identification and staging of pancreatic ductal adenocarcinoma
Authors: Minniti Salvatore , Bruno Costanza , Biasiutti Carlo , Tonel Davide , Falzone Andrea , Falconi Massimo , Procacci Carlo
Published: April 2003

Purpose

The aim of this prospective study was to compare sonography with helical CT in the identification and staging of ductal adenocarcinoma of the pancreas.

Methods

Sixty‐four patients with histopathologically proven pancreatic ductal adenocarcinoma underwent both sonography and helical CT at our hospital between November 1, 2000, and October 31, 2001. These diagnostic imaging examinations were performed by 2 independent radiologists who were unaware of the findings of any other imaging or histopathologic examination and who assessed each case for the presence of tumor, involvement of peripancreatic vessels, the presence of metastases in the liver, and the resectability of the tumor. In the patients who underwent subsequent laparotomy, imaging‐based diagnoses of overall resectability and vascular involvement were compared with surgical findings, which were considered the gold standard. Imaging‐based diagnoses for the presence of hepatic metastases were compared with the findings of intraoperative sonography in patients who underwent radical resection and with the surgical findings in the patients who underwent palliative surgery; the operative findings were considered the gold standard.

Results

In the identification of pancreatic adenocarcinoma, sonography was more accurate (61 of 64 tumors, 95.3%) than helical CT was (57 of 64 tumors, 89.1%). In the overall prediction of resectability among the 43 patients who underwent laparotomy, sonography (81.4%) was less accurate than CT was (86.0%). In diagnosing involvement of the superior mesenteric artery, sonography (95.3%) was more accurate than CT (88.4%). The 2 methods were equally accurate in diagnosing hepatic metastases (86.0%) and involvement of the celiac trunk (100%) and the superior mesenteric vein (88.4%), but sonography was less accurate than CT for diagnosing involvement of the common hepatic artery (90.7% versus 95.3%, respectively) and portal vein (88.4% versus 93.0%, respectively).

Conclusions

Sonography is more reliable than CT in identifying pancreatic adenocarcinoma, and its accuracy in staging is similar to that of CT. © 2003 Wiley Periodicals, Inc. 31:175–182, 2003
Left diaphragmatic eventration diagnosed as congenital diaphragmatic hernia by prenatal sonography
Authors: Yang Jeong In
Published: April 2003

Congenital diaphragmatic eventration is an abnormal elevation of the diaphragm resulting from failure of muscle fibers to develop during gestation. A mediastinal shift to the contralateral side may cause significant compression of the affected chest contents, resulting in compromised pulmonary function, especially when both sides are involved. Differentiating between congenital diaphragmatic eventration and congenital diaphragmatic hernia is very difficult but important because eventration has a better perinatal outcome than diaphragmatic hernia has. This report discusses a case of congenital diaphragmatic eventration that was initially diagnosed by prenatal sonography as a diaphragmatic hernia on the left side. In this case, the correct diagnosis was made at the time of surgery shortly after the infant's birth. The defect was repaired by plication, and the infant's outcome was favorable. Although the initial diagnosis was incorrect in this case, the use of sonography can aid in the prenatal detection and diagnosis of congenital diaphragmatic anomalies. © 2003 Wiley Periodicals, Inc. J Clin Ultrasound 31:214–217, 2003
Seminal vesicle masses detected incidentally during transrectal sonographic examination of the prostate
Authors: Al‐Saeed Osama , Sheikh Mehraj , Kehinde E. O. , Makar R.
Published: April 2003

Purpose

The aim of this study was to review the incidence and type of seminal vesicle (SV) masses discovered during transrectal sonography (TRUS) of the prostate.

Methods

Patients were a consecutive series of men referred for TRUS of the prostate because of lower urinary tract symptoms or elevated prostate‐specific antigen levels, who were found on TRUS to have SV masses. Patients with prostate cancer involving the SVs were excluded. Cystic lesions were sampled by fine‐needle aspiration and solid lesions by core biopsy, all under sonographic guidance.

Results

Of the 450 patients who underwent TRUS between January 1997 and December 2001, 10 (2%) were found to have SV masses; 5 masses were cystic and 5 were solid. Cytologic evaluation of aspirated specimens revealed benign findings in all 5 cysts. Four of the 5 patients with solid SV masses had chronic schistosomiasis, and the mass in the fifth patient was a metastatic deposit from a renal cell carcinoma.

Conclusions

SV masses may be responsible for lower urinary tract symptoms in a small fraction of cases. A cystic mass is presumptively benign, whereas a solid lesion has a small probability of being malignant, especially if the patient has a primary neoplasm elsewhere. Schistosomiasis should always be considered when making a differential diagnosis in patients who live in areas where infestation is endemic. © 2003 Wiley Periodicals, Inc. J Clin Ultrasound 31:201–206, 2003
Use of a curved‐array transducer to reduce interobserver variation in sonographic measurement of thyroid volume in healthy adults
Authors: Peeters Els Y. , Shabana Wael M. , Verbeek Peter A. , Osteaux Michel M.
Published: April 2003

Purpose

Sonographic calculation of thyroid volume is used in the diagnosis and follow‐up of thyroid diseases. Since the calculated volume of thyroid lobes is highly influenced by the longest (ie, craniocaudal) diameter, we examined whether using a curved‐array transducer as opposed to a linear‐array transducer to measure the craniocaudal diameter would reduce interobserver variation.

Methods

Three sonographers with different levels of expertise each used a 5–12‐MHz linear‐array transducer and a 2–5‐MHz curved‐array transducer to measure the craniocaudal diameter of both thyroid lobes of 25 healthy volunteers. On the basis of these measurements, thyroid lobe volumes were calculated. Single‐factor analysis of variance was used to evaluate the interobserver variations between the measurements made by all 3 observers as well as between measurements taken by pairs of observers. A p value of less than 0.05 was considered significant.

Results

Using the linear‐array transducer to measure the craniocaudal diameter resulted in significant interobserver variation in thyroid volume calculation (p = 0.02), whereas using the convex‐array transducer did not. Using either transducer resulted in a highly significant interobserver variation in measurements of the craniocaudal diameter, although the variation was far more pronounced for measurements made with the linear‐array transducer (p = 0.0005) than for those made with the curved‐array transducer (p = 0.04). For both transducers, the interobserver variations were most pronounced between the most and the least experienced sonographers.

Conclusions

To avoid significant interobserver variation in calculating thyroid lobe volume, we recommend using a curved‐array transducer to measure the craniocaudal diameter of the thyroid lobes. © 2003 Wiley Periodicals, Inc. J Clin Ultrasound 31:189–193, 2003
Handheld ultrasound device for detection of non‐opaque and semi‐opaque foreign bodies in soft tissues*
Authors: Levy Angela D. , Harcke H. Theodore
Published: April 2003

Purpose

The aim of this study was to determine whether a portable handheld ultrasound device can be used to visualize non‐opaque and semi‐opaque foreign bodies in soft tissues and to characterize their sonographic appearance.

Methods

Twenty‐two fragments composed of various natural and man‐made materials and ranging from 4 to 20 mm in the maximum longitudinal dimension were embedded into 2 thawed turkey breasts. Two sonographers who were unaware of the number, dimensions, and locations of the fragments used a handheld ultrasound device to detect and characterize the fragments on the basis of their surface echogenicity and acoustic shadowing. The sonographic appearance of each fragment was compared with its actual characteristics.

Results

All 22 embedded fragments were successfully identified and localized on sonography. Surface echogenicity and acoustic shadowing varied among the fragments. For some fragments, the sonographically measured maximum longitudinal dimension differed substantially from the actual dimension.

Conclusions

Use of a portable handheld ultrasound device is effective for the detection of foreign bodies with a diameter greater than 4 mm in soft tissues. The sonographic appearances of non‐opaque and semi‐opaque foreign bodies vary in their patterns of surface echogenicity and acoustic shadowing. © 2003 Wiley Periodicals, Inc. J Clin Ultrasound 31:183–188, 2003
Sonographic measurement of the fetal cerebellum, cisterna magna, and cavum septum pellucidum in normal fetuses in the second and third trimesters of pregnancy
Authors: Serhatlioglu Selami , Kocakoc Ercan , Kiris Adem , Sapmaz Ekrem , Boztosun Yasemin , Bozgeyik Zulkif
Published: April 2003

Purpose

Absence of the cavum septum pellucidum (CSP), the cisterna magna (CM), or both, and enlargement of either or both structures are associated with various central nervous system malformations. In an effort to determine normal sizes and relationships between these cranial structures, we measured the CSP and CM in normal fetuses in the second and third trimesters of pregnancy using transabdominal sonography.

Methods

Women with uncomplicated pregnancies and normal singleton fetuses between 16 and 38 weeks' menstrual age were included in this prospective study. The width and anteroposterior (AP) diameters of the CSP were measured on the transverse transventricular plane, and the AP diameter of the CM was measured on the transcerebellar plane from the posterior aspect of the cerebellar vermis to the inner edge of the cranium. The transverse and AP diameters of the cerebellum were also measured. In addition, we measured the biparietal diameters (BPDs).

Results

In total, 130 women participated; 64 were examined in their second trimester and 66 in their third trimester. The mean age of the women was 27.4 ± 4.8 years (range, 18–38 years), and the mean menstrual age of the fetuses was 26.9 ± 6.7 weeks (range, 16–38 weeks). The mean BPD was 66.8 ± 18.7 mm (range, 30–96 mm). The mean width and AP diameter of the CSP and the mean AP diameter of the CM differed significantly between the second and third trimesters (p < 0.001). All measured parameters correlated significantly with menstrual age and BPD.

Conclusions

In normal fetuses, the CSP and CM should be visible on transabdominal sonography between 16 and 38 weeks' menstrual age. Because abnormalities in these cranial structures may be indicative of central nervous system malformations, the availability of mean sonographic measurements from normal fetuses should be helpful in determining the need for additional testing in fetuses with abnormal measurements. © 2003 Wiley Periodicals, Inc. J Clin Ultrasound 31:194–200, 2003
5‐Fluorouracil efficiently enhanced apoptosis induced by adenovirus‐mediated transfer of caspase‐8 in DLD‐1 colon cancer cells
Authors: Uchida Hiroaki , Shinoura Nobusada , Kitayama Joji , Watanabe Toshiaki , Nagawa Hirokazu , Hamada Hirofumi
Published: April 2003

Background

In order to develop a safe and effective gene therapy for cancer, more powerful therapeutic genes must be selected and a gene transduction methodology needs to be devised that minimizes the total dose of vector required. We investigated the combination effect of 5‐fluorouracil (5‐FU), a first‐choice drug for the treatment of colorectal cancer and adenovirus‐mediated transfer of caspase‐8 in DLD‐1 colon cancer cells.

Methods

The degree of cell death was assessed by determining the percentage of cells which had died, and the degree of DNA fragmentation. The protein expression levels and degree of activation of caspase‐8 were analyzed by Western blot analysis. The degree of transgene expression was assessed using adenoviral vectors expressing lacZ and GFP.

Results

Combination treatment led to a significant induction of apoptosis, whereas treatment with either approach alone resulted in only minimal cytotoxicity. Caspase‐8 was only activated in cells that received the combined treatment. Exposure to 5‐FU increased the quantity of transgene expression per cell, 48 h post‐infection. A potentiating effect of adenoviral treatment was also seen when 5‐FU treatment was substituted by the overexpression of cyclin‐dependent kinase inhibitors, p21WAF1/CIP1 and p27KIP1, suggesting that the cytostatic effect of 5‐FU augmented apoptosis induced by caspase‐8 gene transduction by inhibiting the dilution of gene products associated with cell division.

Conclusions

This combination strategy may be very useful in the treatment of 5‐FU‐resistant colorectal cancers and may also be more generally helpful in minimizing the dose of therapeutic vectors used in cancer gene therapy. Copyright © 2002 John Wiley & Sons, Ltd.
The secretory leukoprotease inhibitor (SLPI) promoter for ovarian cancer gene therapy
Authors: Barker Shannon D. , Coolidge Candace J. , Kanerva Anna , Hakkarainen Tanja , Yamamoto Masato , Liu Bin , Rivera Angel A. , Bhoola Snehal M. , Barnes Mack N. , Alvarez Ronald D. , Curiel David T. , Hemminki Akseli
Published: April 2003

Background

Adenoviruses allow efficient transduction of dividing and non‐dividing cells and their safety for the treatment of cancer has been established in clinical trials. However, one disadvantage is their promiscuous tropism. In this regard, tissue‐specific promoters (TSPs) could be useful for directing transgene expression to target tissues and for reducing adverse effects in non‐target tissues. We hypothesize that selective adenovirus‐mediated transgene expression could be achieved through the use of the secretory leukoprotease inhibitor (SLPI) promoter in the context of ovarian cancer.

Methods

Adenoviruses containing the SLPI promoter driving reporter and suicide gene expression were created and tested in ovarian cancer cell lines and primary tumor cells isolated from patients. To evaluate the in vivo activation of the SLPI promoter in comparison to a ubiquitous promoter, intraperitoneal delivery was performed in tumor‐bearing mice, followed by analysis of survival or gene expression in normal organs and tumor.

Results

The SLPI promoter retained its fidelity in an adenoviral context and was activated in both cell lines and primary cancer cells. The SLPI promoter was induced to a high degree in ovarian cancer cells while showing significantly reduced activity in normal tissues. The therapeutic efficacy of SLPI promoter‐controlled gene expression was similar to the ubiquitous promoter in vitro and in an orthotopic murine model of peritoneally disseminated ovarian cancer, with higher activity than controls.

Conclusions

The SLPI promoter is a potentially useful TSP for ovarian cancer and facilitates further development of targeting strategies for improved gene therapy of ovarian carcinomas. Copyright © 2002 John Wiley & Sons, Ltd.
Conditional cell suicide using dox‐dependent caspase‐2 expression
Authors: Knott Anja , Müller Yvonne , Zunino Susan J. , Berens Christian , Hillen Wolfgang
Published: April 2003

Background

Adoptive immune transfer is used as an efficient treatment modality to achieve a graft‐versus‐leukemia effect in persisting or relapsing residual leukemic disease. Safety considerations dictate the need for equipping the transferred cells with a conditional suicide mechanism to eliminate donor T cells when graft‐versus‐host disease occurs. We have examined in a model system using HeLa cells whether doxycycline (dox)‐dependent expression of pro‐apoptotic proteins could be used as a potential new strategy for conditional cell elimination.

Methods

Four constructs encoding pro‐apoptotic proteins were tested in transient transfections to identify suitable cell death inducers. Murine caspase‐2 placed under Tet‐control was chosen for stable transfection into cell lines carrying different dox‐dependent transregulators. The efficiency of cell death induction and the expression patterns of caspase‐2 were analyzed in the respective clones.

Results

Different levels of induced cell death were obtained depending on the properties of the transregulators used to control target gene expression. High expression levels of caspase‐2 in the presence of dox were required to achieve efficient induction of cell death, while tight repression in the absence of inducer was not necessary for cell survival. Dox treatment for 48 h resulted in 94% cell death indicating a very efficient conditional suicide mechanism.

Conclusions

We propose that the principle of using pro‐apoptotic cellular proteins placed under appropriate dox‐dependent regulation may represent an alternative conditional suicide mechanism to the frequently used herpes simplex virus thymidine kinase (HSV‐TK)/ganciclovir‐system, which harbors immunological and toxicological risks. Copyright © 2002 John Wiley & Sons, Ltd.
Spatial and temporal control of transgene expression in vivo using a heat‐sensitive promoter and MRI‐guided focused ultrasound
Authors: Guilhon E. , Voisin P. , de Zwart J. A. , Quesson B. , Salomir R. , Maurange C. , Bouchaud V. , Smirnov P. , de Verneuil H. , Vekris A. , Canioni P. , Moonen C. T. W.
Published: April 2003

Background

Among the techniques used to induce and control gene expression, a non‐invasive, physical approach based on local heat in combination with a heat‐sensitive promoter represents a promising alternative but requires accurate temperature control in vivo. MRI‐guided focused ultrasound (MRI‐FUS) with real‐time feedback control allows automatic execution of a predefined temperature‐time trajectory. The purpose of this study was to demonstrate temporal and spatial control of transgene expression based on a well‐defined local hyperthermia generated by MRI‐FUS.

Methods

Expression of the green fluorescent protein (GFP) marker gene was used. Two cell lines were derived from C6 glioma cells. The GFP expression of the first one is under the control of the CMV promoter, whereas it is under the control of the HSP70 promoter in the second one and thus inducible by heat. Subcutaneous tumours were generated by injection in immuno‐deficient mice and rats. Tumours were subjected to temperatures varying from 42 to 50 °C for 3 to 25 min controlled by MRI‐FUS and analyzed 24 h after the heat‐shock. Endogenous HSP70 expression and C6 cell distribution were also analyzed.

Results

The results demonstrate strong expression at 50 °C applied during a short time period (3 min) without affecting cell viability. Induced expression was also clearly shown for temperature in the range 44–48 °C but not at 42 °C.

Conclusions

Heating with MRI‐FUS allows a tight and non‐invasive control of transgene expression in a tumour. Copyright © 2002 John Wiley & Sons, Ltd.
Strategic advantages of insulin‐like growth factor‐I expression for cardioprotection
Authors: Chao Wei , Matsui Takashi , Novikov Mikhail S. , Tao Jingzang , Li Ling , Liu Heling , Ahn Youngkeun , Rosenzweig Anthony
Published: April 2003

Background

Insulin‐like growth factor‐I (IGF‐I) peptide has beneficial effects on cardiomyocyte function and survival, many of which are mediated through the serine‐threonine kinase, Akt. However, concerns about systemic effects of IGF‐I peptide limit its clinical application. The present study tested whether local IGF‐I expression could mediate cardioprotection without elevating serum [IGF‐I].

Methods

The ability of a recombinant adenovirus encoding IGF‐IB (Ad.IGF‐I) to activate Akt and protect cardiomyocytes from hypoxia‐induced apoptosis in vitro was compared with the effects of IGF‐I peptide or expression of constitutively active Akt (myr‐Akt). In vivo, cardiac IGF‐I gene transfer was performed prior to ischemia‐reperfusion injury (IRI). Effects on the ischemic and infarcted areas were assessed while serum [IGF‐I] was measured by radioimmunoassay.

Results

Compared with IGF‐I peptide, Ad.IGF‐I achieved more sustained activation of Akt and reduced hypoxia‐induced apoptosis at lower media IGF‐I concentrations. In a co‐culture system, Ad.IGF‐I protected both infected and uninfected cells from hypoxic injury, while myr‐Akt protected only infected cells. In vivo cardiac injection of Ad.IGF‐I mediated significant local IGF‐I expression, without affecting serum [IGF‐I] levels. After IRI, Ad.IGF‐I did not affect the ischemic area but reduced infarct size ∼50% (32 ± 13 vs. 64 ± 14% AAR in Ad.GFP rats, p < 0.003), although the transgene was expressed in only ∼15% of the ischemic region, consistent with possible paracrine benefit.

Conclusions

Somatic gene transfer of IGF‐I may offer strategic advantages over both systemic delivery of IGF‐I peptide and expression of cell autonomous cardioprotective transgenes such as Akt by mediating autocrine and paracrine cardiomyocyte protection without elevating serum [IGF‐I] levels. Copyright © 2002 John Wiley & Sons, Ltd.
Generation of fiber‐modified adenovirus vectors containing heterologous peptides in both the HI loop and C terminus of the fiber knob
Authors: Koizumi Naoya , Mizuguchi Hiroyuki , Utoguchi Naoki , Watanabe Yoshiteru , Hayakawa Takao
Published: April 2003

Background

Fiber‐modified adenovirus (Ad) vectors can be effective in overcoming the limitations of conventional Ad vectors, specifically their inefficient gene transfer into cells lacking the primary receptor, the coxsackievirus and adenovirus receptor (CAR). Several types of fiber‐modified Ad vectors have been developed. In this study, we evaluated the functionality of several fiber‐modified Ad vectors.

Methods

We developed a simple method based on in vitro ligation to construct Ad vectors containing heterologous foreign peptides in both the HI loop and C terminus of the fiber knob. A functional comparison of Ad vectors containing RGD and/or K7 (KKKKKKK) peptide in the HI loop or C terminus of the fiber knob was performed in several types of human, mouse, and rat cells, including CAR‐positive and ‐negative cells, and tumor cells in mice in vivo.

Results

In the case of the in vitro experiment, Ad vectors containing RGD peptide in the HI loop of the fiber knob showed a higher level of gene transfer than vectors containing RGD peptide at the C terminus of the fiber knob. Ad vectors containing K7 peptide at the C terminus of the fiber knob showed levels of gene transfer similar to those of Ad vectors containing RGD peptide in the HI loop of the fiber knob, depending on the cell type. Ad vectors containing both peptides in the HI loop or C terminus of the fiber knob showed the highest levels of gene transfer and a broader tropism. For gene transfer into tumor cells in vivo, the Ad vectors containing RGD peptide were the most efficient.

Conclusions

In the experiment using cultured cells, Ad vectors containing both RGD and K7 peptides were the most efficient with a broader tropism. In contrast, in the experiment in vivo, Ad vectors containing RGD peptide in the HI loop of the fiber knob were more efficient than the vectors containing K7 peptide (including double‐modified vectors containing both the RGD and K7 peptides). These comparative analyses could provide a systemic reference for the use of fiber‐modified Ad vectors. Our simple method, in which the peptide of interest can be expressed in Ad vectors in either the HI loop or the C terminus of the fiber knob, or both, could be a powerful tool for gene transfer into mammalian cells in studies of gene function as well as in gene therapy. Copyright © 2002 John Wiley & Sons, Ltd.
Gene electro‐transfer improves transduction by modifying the fate of intramuscular DNA
Authors: Cappelletti Manuela , Zampaglione Immacolata , Rizzuto Gabriella , Ciliberto Gennaro , Monica Nicola La , Fattori Elena
Published: April 2003

Background

Intramuscular gene delivery through injection of plasmid DNA has long been considered a promising approach for safe and simple in vivo gene expression for vaccination and gene therapy purposes. Recently, intramuscular gene delivery has been improved by applying low‐voltage electric pulses after plasmid injection, a procedure that has been variably called gene electro‐transfer, in vivo electroporation or electrical stimulation. Different types of electrical treatments have been used with excellent results both in terms of transgene expression levels and immunization outcome. This approach, therefore, holds promise for safe gene delivery to animals and humans designed for non‐viral gene therapy and DNA‐based vaccination. The molecular mechanisms underlying this increment in transduction efficiency are, however, still unclear.

Methods

Plasmid DNA status and kinetics following gene electro‐transfer was analyzed by different methods (Southern analysis, Q‐PCR and transformation into competent bacteria).

Results

A large amount of plasmid DNA is degraded in the first 4 h post‐injection, with or without electroporation; later, the amount of intramuscular plasmid DNA is higher in electroporated samples. On electroporation, plasmid is partially protected from degradation, presumably by its early compartmentalization into the nuclei of muscle cells.

Conclusions

By investigating the intracellular outcome and persistence of plasmid DNA following simple injection or gene electro‐transfer we provide useful information on the mechanisms of plasmid entry and expression and underline some of the steps that could be taken to further improve this methodology. Copyright © 2002 John Wiley & Sons, Ltd.
Antibody targeted gene transfer to endothelium
Authors: Tan P. H. , Manunta M. , Ardjomand N. , Xue S. A. , Larkin D. F. P. , Haskard D. O. , Taylor K. M. , George A. J. T.
Published: April 2003

Background

One of the drawbacks of the currently available vectors for gene therapy is the lack of selectivity in gene delivery. We have therefore investigated a strategy to generate immunoliposomes to target non‐viral vectors to cell surface receptors on endothelium.

Materials and methods

We have developed a novel method of coupling antibodies (Abs) to liposomes complexed to DNA, using mild heat treatment to aggregate the immunoglobulin G (IgG). The interaction of plasmid DNA, liposomes and Abs was measured using a gel retardation assay and a resonant mirror biosensor. The size of the transfection complex was determined by light scattering, and the binding and internalization of the complex to cells was followed using flow cytometry. The transfection ability was tested on cell lines and primary cells in vitro and human corneal or vascular tissues ex vivo.

Results

The interaction of antibodies with liposomes is relatively stable (t1/2 ≅ 45 min). The size of the liposome, Ab and DNA complex was found to be around 500 nm in 4% BSA. The addition of anti‐transferrin receptor Abs increased the internalization of the liposome‐DNA complex into cells. Abs against both transferrin receptor and E‐selectin were shown to augment transfection efficiency of liposomes to cell expressing the appropriate antigens. They are also shown to be efficient in mediating gene delivery to corneal and vascular tissues ex vivo.

Conclusions

We have shown that our novel vector is capable of in vitro and ex vivo gene delivery to cells and human tissues including cornea, artery and vein. In particular, an Ab against E‐selectin was effective at selectively delivering genes to activated endothelial cells expressing the adhesion molecule. Such a strategy will have applications for targeting these tissues prior to transplantation or autologous grafting, and, in the longer term, may allow in vivo targeting of gene therapy to inflammatory sites. Copyright © 2002 John Wiley & Sons, Ltd.
Generation of dendritic cells from lentiviral vector‐transduced CD34+ cells from HIV+ donors
Authors: Gruber A. , Chen I. , Kuhen K.L. , Wheat J.C. , Law P. , Wong‐Staal F.
Published: April 2003

Dendritic cells hold promise as adjuvant for immunotherapy for cancer and infectious diseases. We demonstrate that a significant number of cryopreserved peripheral blood CD34+ cells from HIV‐infected subjects can be transduced with a replication‐incompetent lentiviral vector expressing HIV antigens. In addition, untransduced and transduced CD34+ cells from HIV‐infected individuals were able to differentiate into dendritic cells with strong T‐cell stimulatory capacity. Thus, cryopreserved CD34+ cells from HIV‐infected subjects may prove useful for immunotherapy for HIV/AIDS. J. Med. Virol. 70: 183–186, 2003. © 2003 Wiley‐Liss, Inc.
Sequential changes in hepatitis A virus genotype distribution in Estonia during 1994 to 2001
Authors: Tallo Tatjana , Norder Helene , Tefanova Valentina , Ott Kristi , Ustina Valentina , Prukk Tiina , Solomonova Olga , Schmidt Jelena , Zilmer Kai , Priimägi Ludmilla , Krispin Tõnu , Magnius Lars O.
Published: April 2003

Hepatitis A virus (HAV) isolates from a large outbreak and from non‐outbreak cases in Estonia were characterized by sequencing the aminoterminal VP1 region. From January 1998 to December 1999, a total of 1,084 cases of hepatitis A were reported to the Harjumaa‐Tallinn and Ida‐Virumaa Health Protection Services in Estonia. The attack rate was highest among males aged 15–29. Initial cases were noted to be associated with injecting drug use. IgM anti‐HAV positive sera were available from 107 hospitalized outbreak cases and from 68 patients sampled during 1994 to 2001. HAV RNA was detected in 42% of sera from 1994–1996 and in 88% of sera from 1998–2001. It was possible to obtain HAV sequences from 83 outbreak and 29 background cases. The outbreak strain was represented by five different sequences, all belonging to subtype IIIA. During the outbreak, this IIIA strain also spread into the general population. All available non‐outbreak isolates from 1994 to 2001 but one belonged to genotype IA and formed distinct clusters as compared to isolates from other parts of the world. One subtype IIIA isolate from 1995 was unrelated to the outbreak strain. Subtype IA had been dominating in Estonia during 1994–2001, but the outbreak strain from 1998 to 1999 was IIIA. This subtype was encountered previously in addicts in Sweden during the 1980s and in Norway at the end of the 1990s. This study supports the use of limited sequencing within the aminoterminal VP1 region for studying the molecular epidemiology of hepatitis A. J. Med. Virol. 70: 187–193, 2003. © 2003 Wiley‐Liss, Inc.
Perinatal and intrafamily transmission of hepatitis B virus in three generations of a low‐prevalence population
Authors: Ördög Katalin , Szendrôi Andrea , Szarka Katalin , Kugler Zoltán , Csire Márta , Kapusinszky Beatrix , Xie Jiuru , Csizmadia Katalin , Brojnás Judit , Rusvai Erzsébet , Tempfli Ágnes , Berencsi György
Published: April 2003

Family members of 47 hepatitis B virus (HBV)‐carrier pregnant women were tested for the presence of hepatitis B surface antigen (HBsAg), other markers of HBV infection, and hepatitis A virus (HAV) antibodies. Eleven members of six families were found to be HBV DNA positive. Five of the anti‐HBe‐positive persons were found to be HBV DNA carriers, too. The mean age of the HBV DNA carriers was found to be lower than that of Hbe carriers; therefore, it is suggested that seroconversion to HBe occurs before the resolution of HBV DNA carrier state. Superinfection with hepatitis A virus was not found to influence the elimination of HBV‐carrier state, as there was no correlation found between the hepatitis A exposure and the hepatitis B virus markers in the families. The low HBV prevalence in the population (0.3%) was in contrast to the high prevalence of the families of the HBV‐carrier mothers (27.1%) and family members with HBV markers (50.4%). Significant positive correlation was found in the proportion of HBV‐positive children, and the HBV history of their parents. When fathers were shown to be seronegative, the probability of HBV transmission was reduced by a factor of 6 (12.5% instead of 75%) probably due to reduced viral load and possibly by other factors. Several results indicate, that the noncytocidal hepatitis B virus clearing mechanism suggested by Guidotti et al. [1996, 1999] was effective also in the HBV‐carrier human population. J. Med. Virol. 70: 194–204, 2003. © 2003 Wiley‐Liss, Inc.
Visual gene diagnosis of HBV and HCV based on nanoparticle probe amplification and silver staining enhancement
Authors: Wang Ye‐Fu , Pang Dai‐Wen , Zhang Zhi‐Ling , Zheng Hu‐Zhi , Cao Jun‐Ping , Shen Jun‐Tao
Published: April 2003

A visual gene‐detecting technique using nanoparticle‐supported gene probes is described. With the aid of gold nanoparticle‐supported 3′‐end–mercapto‐derivatized oligonucleotide serving as detection probe, and 5′‐end –amino‐derivatized oligonucleotide immobilized on glass surface acting as capturing probe, target DNA was detected visually by sandwich hybridization based on highly sensitive “nano‐amplification” and silver staining. Different genotypes of Hepatitis B and C viruses in the serum samples from infected patients were detected using home‐made HBV, HCV, and HBV/HCV gene chips by the gold/silver nanoparticle staining amplification method. The present visual gene‐detecting technique may avoid limitations with the reported methods, for its high sensitivity, good specificity, simplicity, speed, and cheapness. This technique has potential applications in many fields, especially in multi‐gene detection gene chips coupled with the detection will find applications in clinic. Additionally, resonance Rayleigh light scattering (RLS) spectroscopy is used, for the first time, to judge and monitor the immobilization of gene probes on gold nanoparticle surfaces. J. Med. Virol. 70: 205–211, 2003. © 2003 Wiley‐Liss, Inc.
Diversification of hypervariable region 1 of hepatitis C virus after liver transplantation
Authors: Fan Xiaofeng , Di Bisceglie Adrian M.
Published: April 2003

Liver transplantation represents a unique model for studying hepatitis C virus (HCV) evolution. In the current study, we estimated the dynamics of HCV quasispecies evolution over a period of 2.5 years both in sera and in tissues, including three different anatomic sites of the liver in a patient undergoing liver transplantation. The patient displayed an extremely homologous virus population in either serum or liver at each time point after liver transplantation. However, there was a successive diversification of hypervariable region 1 (HVR1) of HCV under such an im‐munocompromised status and the preexistence of matching HVR1 antibodies. This distinct evolutionary pattern of HVR1 suggests that immunosuppressed status does contribute to diminished genetic diversity of HVR1 at a given time point after liver transplantation. However, the dramatic diversification of HVR1 sequence over time is mainly due to the adaptation of new replicative site, the donor liver, indicating that HCV may obtain the higher replication fitness by mutating HVR1 domain. J. Med. Virol. 70: 212–218, 2003. © 2003 Wiley‐Liss, Inc.
Upregulation of endogenous intrahepatic interferon stimulated genes during chronic hepatitis C virus infection
Authors: MacQuillan Gerry C. , Mamotte Cyril , Reed William D. , Jeffrey Gary P. , Allan Jane E.
Published: April 2003

The success of interferon‐α and ribavirin combination therapy for the treatment of chronic hepatitis C viral infection differs between patients. In an attempt to identify predictors of host response to therapy, the levels of mRNA for interferon (IFN) stimulated genes: MxA, PKR, 2′5′ OAS, ISG15, and interleukin 8 (IL‐8), were examined in liver by real‐time RT‐PCR prior to commencement of therapy. The levels of intrahepatic classical IFN stimulated genes, but not IL‐8, in chronic HCV disease (n = 44) were found to be significantly upregulated (P < 0.001) compared to the control cohort (n = 12). The genotype of the infecting HCV strain did not influence IFN stimulated gene expression. These results suggest that the endogenous type 1 IFN antiviral effector pathway is broadly activated during chronic HCV disease, although the levels of mRNA for any of the IFN‐stimulated genes tested did not predict the outcome of combination therapy. J. Med. Virol. 70: 219–227, 2003. © 2003 Wiley‐Liss, Inc.
Rapid and quantitative detection of human adenovirus DNA by real‐time PCR
Authors: Heim Albert , Ebnet Carmen , Harste Gabi , Pring‐Åkerblom Patricia
Published: April 2003

Rapid diagnosis of human adenovirus (HAdV) infections was achieved by PCR in the recent years. However, conventional PCR has the risk of carry‐over contamination due to open handling with its products, and results are only qualitative. Therefore, a quantitative “real‐time” PCR with consensus primer and probe (dual fluorescence labelled, “TaqMan”) sequences for a conserved region of the hexon gene was designed and evaluated. Real‐time PCR detected all 51 HAdV prototypes. Sensitivity of the assay was ≤15 copies/run and the linear range of quantitation 1.5 × 101 to 1.5 × 108 copies/run. TaqMan PCR gave identical results compared to an established conventional one‐step PCR protocol in 218 (38 positive and 180 negative) of 234 clinical samples including blood, serum, eye swabs, and feces, and had divergent results in 16 samples (15 positive only in TaqMan PCR, all with low copy numbers, and one positive only in conventional PCR), indicating a higher sensitivity of TaqMan PCR. Adenovirus viremia was detected by TaqMan PCR in 4 of 27 (14.8%) paediatric and 8 of 93 (8.6%) adult stem cell transplant recipients but only in 5 of 306 healthy controls (blood donors, 1.6%). Virus loads of pediatric patients (median 1.7 × 105) were significantly higher than in adult patients (median 2.3 × 103) and than in controls (all samples ≤1.7 × 103 copies/ml). A few immunosuppressed children had very high virus loads (up to 1.1 × 1010 copies/ml), which were associated with symptoms of disseminated disease. In conclusion, real‐time PCR is a sensitive and quantitative procedure for the detection of adenovirus infections. J. Med. Virol. 70: 228–239, 2003. © 2003 Wiley‐Liss, Inc.
Single human cytomegalovirus gB genotype shed in multiple sites at the time of diagnosis in renal transplant recipients
Authors: Carraro E. , Granato C.F.H.
Published: April 2003

Human cytomegalovirus (HCMV) is a major cause of morbidity and mortality in immunocompromised patients, such as renal transplant recipients. Analysis of the gene encoding the envelope glycoprotein B (gB) showed that clinical isolates adopted one of the sequence configurations, permitting the isolates to be assigned a gB genotype of 1–4. It has been suggested that HCMV gB genotypes could be correlated with tropism and pathogenesis. A number of reports in the literature refer to shedding of different gB strains, permitting follow‐up of renal transplant recipients. Considering that a single strain might be responsible for the clinical expression of the disease in multiply exposed individuals, the frequency distribution of gB genotypes was examined by nested polymerase chain reaction and restriction fragment length polymorphism in 20 renal transplant recipients at the time of diagnosis. The association between gB genotypes and cellular tropism was determined using blood, saliva, and urine for each patient. HCMV gB genotype 2 was found more frequently than other genotypes (gB2, 40%; gB1, 30%; gB3, 25%; and gB4, 5%) in renal transplant recipients. The gB type did not correlate with tropism for different body sites. All the patients with HCMV infections presumably harbored a single HCMV strain at the time of diagnosis. In multiply exposed patients, the immunomodulation provided by acute HCMV infection could favor later shedding of different strains. J. Med. Virol. 70: 240–243, 2003. © 2003 Wiley‐Liss, Inc.
Contrasting effects of hydroxyurea on cell growth and reduction in Epstein‐Barr virus genomes in EBV‐infected epithelioid cell lines vs. Burkitt's lymphoma cell lines
Authors: Jiang Ru , Kanamori Mikiko , Satoh Yukio , Fukuda Makoto , Ikuta Kazufumi , Murakami Masanao , Sairenji Takeshi
Published: April 2003

Eliminating Epstein‐Barr virus (EBV) genomes from infected cells is an intriguing theoretical strategy in therapy for EBV‐associated malignant diseases. Respective patterns were characterized for hydroxyurea (HU)‐promoted loss of EBV genomes from EBV‐infected epithelioid cell lines derived from the noncancerous portion of gastric carcinoma tissues and Burkitt's lymphoma (BL) cell lines. Epithelioid cell lines GT38 and PN were less sensitivity to HU than BL cell lines Akata, Raji, and Daudi in terms of cell growth inhibition and cell cycle arrest. On passage in medium with 50 μM HU, the fraction of EBV nuclear antigen (EBNA)‐positive cells was reduced substantially in the BL cell lines, but only slightly in the epithelioid cell lines. EBV DNA was reduced in Akata, Raji, and Daudi cells upon passage in 50 μM HU by 95%, 70%, and 50%, respectively, but by only 10% in GT38 cells, in which EBV DNA reduction was enhanced at increased concentrations of HU. This indicates that EBV genome is more easily lost from BL cell lines than from epithelioid cell lines upon culturing in HU. These findings support the view that the elimination of EBV could be therapeutically effective in EBV‐associated BL by HU. J. Med. Virol. 70: 244–252, 2003. © 2003 Wiley‐Liss, Inc.
Infectious mononucleosis patients temporarily recognize a unique, cross‐reactive epitope of Epstein‐Barr virus nuclear antigen‐1
Authors: McClain Micah T. , Rapp Erin C. , Harley John B. , James Judith A.
Published: April 2003

The spectrum of antibodies against Epstein‐Barr virus nuclear antigen‐1 (EBNA‐1) in patients with a recent history of infectious mononucleosis and nonaffected EBV‐positive individuals has been characterized by epitope mapping. Sera were evaluated for antibodies to all unique maximally overlapping octapeptides of EBNA‐1. Both normal controls and patients with infectious mononucleosis produce IgG antibodies that recognize the glycine‐alanine‐rich portion of EBNA‐1, as previously described. All EBNA‐1 IgG‐positive infectious mononucleosis patients tested, however, consistently produce IgG specific for an additional epitope (aa 398–412 PPPGRRPFFHPVGEA) near the middle of the EBNA‐1 protein. This region was not found to be antigenic in healthy EBV‐seropositive individuals. This region does, however, cross‐react with the sequence PPPGMRPP from the common lupus spliceosomal autoantigen Sm B′ in several infectious mononucleosis patient sera. Patients with recent clinical infectious mononucleosis temporarily recognize a unique cross‐reactive epitope of EBNA‐1 not bound by antibodies from non‐infectious mononucleosis EBV‐positive sera or those with a distant history of IM. J. Med. Virol. 70: 253–257, 2003. © 2003 Wiley‐Liss, Inc.
Infantile viral gastroenteritis: On the way to closing the diagnostic gap
Authors: Simpson R. , Aliyu S. , Iturriza‐Gómara M. , Desselberger U. , Gray J.
Published: April 2003

A total of 305 faecal specimens collected from children under the age of 5 who presented with symptoms of acute gastroenteritis either as inpatients at Addenbrooke's Hospital (N = 100) or to General Practitioners in East Anglia (N = 205) during 1999–2001 were tested for the presence of rotavirus, norovirus, sapovirus, enteric adenoviruses (Group F, serotypes 40 and 41), and astrovirus. An aetiologic agent was found in 184 specimens (60.3%). The most commonly found single viral pathogen was rotavirus (27.9%), followed by norovirus (13.4%), enteric adenoviruses (7.9%), astrovirus (2.3%), and sapovirus (1%). Mixed infections were observed in 27 specimens (8.9%), and no aetiologic agent was found in over a third of the specimens tested. These data demonstrate that the diagnostic gap can be reduced considerably through the use of molecular amplification and detection techniques. However, additional work is required to reduce this deficit further by optimising sampling algorithms and by identifying other agents associated with viral gastroenteritis. J. Med. Virol. 70: 258–262, 2003. © 2003 Wiley‐Liss, Inc.
Intradermal doxorubicin reduces ganglionic reactivation of latent herpes simplex virus in mice after pretreatment with hypertonic saline
Authors: Sekizawa Tsuyoshi , Openshaw Harry , Itoyama Yasuto
Published: April 2003

Recurrent lesions from herpes simplex virus (HSV) occur after reactivation of latent HSV in neurons of sensory ganglion, axonal transport of reactivated virus, and HSV replication on the skin. A potential treatment strategy is to inject epithelial sites of frequent recurrences with anti‐viral or cytotoxic agents that are taken up by nerve terminals and transported by axoplasmic flow to latently infected ganglionic neurons. Doxorubicin is transported by nerves and destroys the corresponding nerve cell bodies, but earlier attempts in HSV animal models required intraneural injection to eliminate HSV infection and this treatment destroyed the nerve and large portions of the ganglion. The present study used intradermal doxorubicin in latently infected mice that had been inoculated with HSV by the lip route and passively immunized at the time of inoculation. As found previously, doxorubicin injection in the lip 2 or more months after HSV inoculation did not eliminate HSV latency as evaluated by recovery of virus from trigeminal ganglionic explants. However, when hypertonic saline was injected in the same site 24 hr prior to doxorubicin, there was a 55% reduction of positive ganglionic explant cultures. Edema from the hypertonic saline may increase access of doxorubicin to nerve terminals. This technique with hypertonic saline, which has also been used to enhance virulence of HSV skin innoculation, may have general application of increasing axoplasmic transport of drugs or biologicals. Skin toxicity may preclude doxorubicin use for HSV recurrences in patients; however, the results of this study support the concept of anti‐HSV treatment via retrograde axoplasmic transport. J. Med. Virol. 70: 263–266, 2003. © 2003 Wiley‐Liss, Inc.
Evaluation of active human herpesvirus 6 infection by reverse transcription‐PCR
Authors: Yoshikawa Tetsushi , Akimoto Shiho , Nishimura Naoko , Ozaki Takao , Ihira Masaru , Ohashi Masahiro , Morooka Masashi , Suga Sadao , Asano Yoshizo , Takemoto Masaya , Nishiyama Yukihiro
Published: April 2003

Monitoring of active human herpesvirus 6 (HHV‐6) infection is important for distinguishing between reactivation and latency of the virus. The reverse transcription polymerase chain reaction (RT‐PCR) may be a useful tool in order to distinguish active and latent HHV‐6 infection. An RT‐PCR assay detecting 4 different HHV‐6 gene transcripts was established. Samples of peripheral blood mononuclear cells (PBMCs) were collected from patients with exanthem subitum and used to evaluate the reliability of the assay. After confirming the reliability of the assay, RT‐PCR was used to determine whether HHV‐6 reactivation occurs in children with hypercytokinemia. Three gene transcripts (U31, U39, and U94) were detected in 90–100% of the PBMC samples collected from febrile period of exanthem subitum patients, from which HHV‐6 was isolated. The two gene transcripts encoding the late proteins U31 and U39, however, were not detected in samples collected during the convalescent period that contained no infectious virus. The putative latency associated gene transcript, U94, was detected in 2 (10%) of the 20 convalescent samples, and another immediate early gene transcript, U90, was also detected in 3 (15%) of the 20 convalescent samples. The frequency of HHV‐6 reactivation in patients with hypercytokinemia, suggesting monocyte/macrophage activation, was studied. Only 9 of 17 patients diagnosed with Kawasaki disease and 1 patient diagnosed with juvenile rheumatoid arthritis were positive for HHV‐6 DNA in their PBMCs samples. Neither the U31 gene nor the U94 gene transcript was detected in any of the 10 samples. An RT‐PCR assay screening for both immediate early and late genes may be useful for monitoring active HHV‐6 infection. No HHV‐6 reactivation was found in patients with hypercytokinemia using the RT‐PCR assay. J. Med. Virol. 70: 267–272, 2003. © 2003 Wiley‐Liss, Inc.