bluuebunny/crossref_metadata_2025 huggingface dataset:
Differential effect of insulin and elevated glucose level on adenosine handling in rat T lymphocytes
Authors: Pawelczyk Tadeusz , Sakowicz‐Burkiewicz Monika , Kocbuch Katarzyna , Szutowicz Andrzej
Published: September 2005

Reduced proliferation potential is among other T cell functional defects long known feature of diabetes. However, the mechanism responsible for this impairment is still unknown. Our study was undertaken to investigate the effect of changes in glucose and insulin concentrations on adenosine metabolism, transport and receptor‐mediated action in rat T lymphocytes. Presented results indicate that vulnerability of T cells to metabolic stress is determined by insulin but not by glucose concentration. However, glucose and insulin differentially affected the activities of adenosine metabolizing enzymes in resting and proliferating T cells. The Con A‐induced proliferation of cultured T lymphocytes did not depended on expression level and functional state of nucleoside transporters. Inhibition of adenosine kinase (AK) with 5‐iodotubercidin lowers the proliferation potential of T cells to the level observed for insulin‐deprived cells. Moreover, insulin‐deprived T lymphocytes but not cells cultured in the presence of insulin released significant quantities of adenosine. Under resting conditions, the cAMP level was fivefold higher in cells deprived of insulin comparing to cells cultured in the presence of insulin. Exposition of insulin‐deprived T lymphocytes to specific antagonist (ZM241385) of A2a receptor but not to specific antagonist (Alloxazine) of A2b receptor suppressed cAMP elevation and completely restored the proliferation potential of T cells. Concluding, adenosine released by insulin‐deprived T cells due to suppressed AK activity by acting on A2a receptors leads to increases in cAMP level and suppression of T cell proliferation. We assume that this mechanism may significantly contribute to immune impairment observed in diabetes. J. Cell. Biochem. © 2005 Wiley‐Liss, Inc.
Neural expression ofα‐internexinpromoter in vitro and in vivo
Authors: Wang Pei , Wang Seu‐Mei , Hsieh Chia‐Ju , Chien Chung‐Liang
Published: September 2005

α‐Internexin is a 66 kDa neuronal intermediate filament protein found most abundantly in the neurons of the nervous systems during early development. To characterize the function of mouseα‐internexinpromoter, we designed two different expression constructs driven by 0.7 kb or 1.3 kb of mouseα‐internexin5′‐flanking sequences; one was the enhanced green fluorescent protein (EGFP) reporter for monitoring specific expression in vitro, and the other was thecrefor studying the functional DNA recombinase in transgenic mice. After introducing DNA constructs into non‐neuronal 3T3 fibroblasts and a neuronal Neuro2A cell line by lipofectamine transfection, we observed that the expression of EGFP with 1.3 kb mouseα‐internexinpromoter was in a neuron‐dominant manner. To establish a tissue‐specific pattern in the nervous system, we generated a transgenic mouse line expressing Cre DNA recombinase under the control of 1.3 kbα‐Internexinpromoter. The activity of the Cre recombinase at postnatal day 1 was examined by mating thecretransgenic mice to ROSA26 reporter (R26R) mice with knock‐in Cre‐mediated recombination. Analyses of postnatal day 1 (P1) newborns showed that β‐galactosidase activity was detected in the peripheral nervous system (PNS), such as cranial nerves innervating the tongue and the skin as well as spinal nerves to the body trunk. Furthermore, X‐gal‐labeled dorsal root ganglionic (DRG) neurons showed positive for α‐Internexin in cell bodies but negative in their spinal nerves. The motor neurons in the spinal cord did not exhibit any β‐galactosidase activity. Therefore, thecretransgene driven by mouseα‐internexinpromoter, described here, provides a useful animal model to specifically manipulate genes in the developing nervous system. J. Cell. Biochem. 97: 275–287, 2006. © 2005 Wiley‐Liss, Inc.
Roles of osteonectin in the migration of breast cancer cells into bone
Authors: Campo McKnight Dianalee A. , Sosnoski Donna M. , Koblinski Jennifer E. , Gay Carol V.
Published: September 2005

The focus of this study was to gain insight into the role(s) of osteonectin in the preferential metastasis of breast cancer cells to bone. Osteonectin was isolated from conditioned media of several cell lines including breast cancer (MDA‐MB‐435, MDA‐MB‐468), osteoblasts (hFOB1.19), non‐neoplastic breast epithelial (hTERT‐HME1), and vascular endothelial cells isolated from a bone biopsy (HBME‐1). Chemical/physical properties of osteonectin from these five sources was analyzed to determine if unique configurations of osteonectin exist and therefore identify a chemotactic isoform. Osteonectin from all sources had a molecular weight of ∼46 kDa, N‐linked glycosylation, and undetectable phosphorylated serines, sialic acids and O‐linked oligosaccharides. The cDNA for osteonectin from the breast cancer, osteoblast, and breast epithelial cell lines was identical, while the vascular endothelial cell cDNA contained point mutations that resulted in eight amino acid substitutions. Bone‐derived osteonectin was then analyzed to assess its influence on breast cancer cell motility and migration. Although osteonectin increased undirected MDA‐MB‐231 cell motility, it did not chemoattract the same breast cancer cell line. However, the breast cancer cells did migrate toward the known chemoattractant vitronectin and to bone extracts derived from wild‐type and osteonectin‐null mice. Migration to vitronectin was enhanced when osteonectin was also present. We concluded that osteonectin was not a chemotactic factor. However, through its anti‐adhesive properties, osteonectin induced undirected breast cancer cell motility, and may have enhanced chemoattraction to vitronectin. J. Cell. Biochem. © 2005 Wiley‐Liss, Inc.
Gender acts as a context for interpreting diagnostic criteria
Authors: Flanagan Elizabeth H. , Blashfield Roger K.
Published: September 2005

This study used a unique methodology to determine the aspects of case vignettes that elicited an effect of case gender on diagnosis. A total of 99 psychiatrists and psychologists were shown cases representing a man or a woman that contained varying numbers of histrionic and antisocial criteria. The cases were presented by computer a few sentences at a time. Clinicians were asked to offer a diagnosis for the case after each group of sentences was presented. Results indicated that case gender tended to affect clinicians' diagnostic decisions when criteria related to that diagnosis were shown, suggesting that case gender was acting as a context in which diagnostic criteria were interpreted. © 2005 Wiley Periodicals, Inc. J Clin Psychol 61: 1485–1498, 2005.
What's an integrationist? A study of self‐identified integrative and (occasionally) eclectic psychologists
Authors: Norcross John C. , Karpiak Christie P. , Lister Kelly M.
Published: September 2005

We examined the views and practices of self‐identified eclectic and integrative psychologists, particularly in the context of historical changes since 1977 and 1988. Results from 187 eclectic clinical psychologists indicated that 50% previously adhered to another theoretical orientation, the majority preferred the term integrative to eclectic, and 85% conceptualized eclecticism/integration as the endorsement of a broader orientation. The most common paths toward integration were theoretical integration, common factors, and assimilative integration, as opposed to technical eclecticism. The most frequent theoretical contributor to integrative practice was cognitive therapy. © 2005 Wiley Periodicals, Inc. J Clin Psychol 61: 1587–1594, 2005.
Brief scheduled phone support from a clinician to enhance computer‐aided self‐help for obsessive‐compulsive disorder: Randomized controlled trial
Authors: Kenwright Mark , Marks Isaac , Graham Candida , Franses Abigail , Mataix‐Cols David
Published: September 2005

Treatment‐resistant obsessive‐compulsive disorder (OCD) patients from around the United Kingdom who employed computer‐guided self‐help by using BTSteps over 17 weeks were randomized to have brief live phone support from a clinician either (1) in nine Scheduled clinician‐initiated calls or (2) only in calls Requested by the patient (n = 22 per condition). Call content and mean duration were similar across conditions. Scheduled‐support patients dropped out significantly less often, did more homework of self‐exposure and self‐imposed ritual prevention (95% vs. 57%), and showed more improvement in OCD symptoms and disability. Mean total support time per patient over 17 weeks was 76 minutes for Scheduled and 16 minutes for Requested patients. Giving brief support proactively by phone enhanced OCD patients' completion of and improvement with computer‐aided self‐help. © 2005 Wiley Periodicals, Inc. J Clin Psychol 61: 1499–1508, 2005.
Reevaluating the distinction between Axis I and Axis II disorders: The case of borderline personality disorder
Authors: Ruocco Anthony C.
Published: September 2005

The division between Axis I clinical syndromes and Axis II personality disorders is a long‐standing distinction based primarily on three guiding principles: phenomenology, cause, and course. Clinical syndromes were generally thought to be characterized by transient symptoms with biological causes and an unstable course; personality disorders were supposed by many to be characterized by long‐standing personality traits, whose roots were primarily psychological, and a stable and unremitting course. Borderline personality disorder (BPD), however, is a condition characterized by distinct clinical symptoms, varied causes, and a relatively unstable course. Past theorizing about the distinction between Axis I and Axis II disorders is presented in light of recent empirical evidence refuting the rationalization for the separation of personality disorders and clinical syndromes using BPD as a means for comparison. © 2005 Wiley Periodicals, Inc. J Clin Psychol 61: 1509–1523, 2005.
Downstream processing of 1,3‐propanediol fermentation broth
Authors: Hao Jian , Xu Feng , Liu Hongjuan , Liu Dehua
Published: September 2005

The downstream processing of 1,3‐propanediol fermentation broth using flocculation, reactive extraction, and reactive distillation was studied. Cellular debris and soluble protein in the broth were flocculated by combined use of chitosan and polyacrylamide at optimal concentrations of 150 ppm and 70 ppm, respectively; the soluble protein in the broth decreased to 0.06 g L−1, and the recovery ratio of the supernatant liquor to broth was greater than 99%. 1,3‐Propanediol and other alcohols were extracted from the supernatant liquor by reacting with butyraldehyde. In a four‐stage countercurrent extraction with the volume ratio of the extraction solvent to the aqueous phase being 20:100, more than 99% 1,3‐propanediol acetal (2‐propyl‐1,3‐dioxane) and 2,3‐butanediol acetal (2‐propyl‐4,5‐dimethyl‐1,3‐dioxolane) were recovered from the aqueous phase; 35% of the glycerol acetals were recovered. The acetals produced were hydrolyzed in a reactive distillation column using the strongly acidic cation‐exchange resin as catalyst, the bottom product obtained was a mixture of 1,3‐propanediol (407 g L−1), 2,3‐butanediol (252 g L−1), glycerol (277 g L−1), and glycerol acetals (146 g L−1). Copyright © 2005 Society of Chemical Industry
Types of vascular wall as related to vasa vasorum in common tree shrew (Tupaia glis)
Authors: Chuncher Sununta , Somana Reon
Published: September 2005

Blood vessels of the common tree shrew (Tupaia glis) were embedded in Araldite, sectioned at 0.5–1 μm thickness, and observed with light microscope (LM). It was found that the vascular wall could be classified into three categories: (1) those with proper vasa vasorum (PVV); (2) those with collaborative vasa vasorum in perimural tissues; (3) those without vasa vasorum. The PVV were located in the the tunica media, between the tunica media and the tunica adventitia, and in the tunica adventitia of the vascular wall. On studying the vascular casts with a scanning electron microscope (SEM), it was noted that what appeared to be vasa vasorum did not always conform to those found under the LM. Furthermore, the PVV seen in a particular blood vessel among different tree shrews were not always related to intraluminal partial pressure of oxygen, vascular size, and thickness of the wall. In contrast, the results of this study indicate that the activeness of the vascular wall in varying the amount of blood flow to certain organs is an important factor that is associated with the existence, density, and distribution of the vasa vasorum. Microsc. Res. Tech. 67:317–324, 2005. © 2005 Wiley‐Liss, Inc.
Cell and matrix morpho‐functional analysis in chondrocyte micromasses
Authors: Battistelli Michela , Borzì Rosa Maria , Olivotto Eleonora , Vitellozzi Roberta , Burattini Sabrina , Facchini Andrea , Falcieri Elisabetta
Published: September 2005

Micromass cultures represent a convenient means of studying chondrocyte physiology in the context of a tridimensional culture model. In this study, we present the first ultrastructural analysis of the distribution and organization of the extracellular components in micromasses in comparison with their cartilaginous counterparts. Primary chondrocytes obtained from osteoarthritis patients were pelleted in micromasses. Transmission electron microscopy and immunofluorescence were used to evaluate the distribution of major extracellular matrix proteins, i.e., aggrecan, chondroitin‐4‐sulfate, chondroitin‐6‐sulfate, and collagen I and II. Both approaches revealed a number of morphological features shared by micromass and cartilage chondrocytes. In particular, in micromasses, chondrocytes are in close contact with an organized extracellular matrix that adequately mimics that of cartilage. Cells were observed to establish specialized junctions for cell–extracellular matrix crosstalk. Noteworthy, cells seem endowed in a chondroitin sulfate‐rich microenvironment, and thus possibly ensuring the immobilization of chemokines, a family of molecules emerging in osteoarthritis pathogenesis, in a haptotactic‐like gradient to the chondrocytes, which facilitates the binding to their receptors. To determine the suitability of this model to investigate osteoarthritis pathogenesis, a potential apoptotic stimulus (endothelial IL‐8) was used, and ultrastructural analysis assessed apoptosis induction. Micromass cultures were proved to be an experimental technique providing a large number of properly differentiated chondrocytes, and thus allowing reliable biochemical and morphological studies. They represent, therefore, a novel approach to osteoarthritis investigation that promises more thorough understanding of chondrocyte physiology in osteoarthritis. Microsc. Res. Tech. 67:286–295, 2005. © 2005 Wiley‐Liss, Inc.
Histochemical evidences on the chronological alterations of the hypertrophic zone of mandibular condylar cartilage
Authors: Hossain Kazi Sazzad , Amizuka Norio , Ikeda Nobuyki , Nozawa‐Inoue Kayoko , Suzuki Akiko , Li Minqi , Takeuchi Kiichi , Aita Megumi , Kawano Yoshiro , Hoshino Masaaki , Oda Kimimitsu , Takagi Ritsuo , Maeda Takeyasu
Published: September 2005

The hypertrophic chondrocytes lack the ability to proliferate, thus permitting matrix mineralization as well as vascular invasion from the bone in both the mandibular condyle and the epiphyseal cartilage. This study attempted to verify whether the histological appearance of the hypertrophic chondrocytes is in a steady state during postnatal development of the mouse mandibular condyle. Type X collagen immunohistochemistry apparently distinguished the fibrous layer described previously as the “articular zone,” “articular layer,” and “resting zone” from the hypertrophic zone. Interestingly, the ratio of the type X collagen‐positive hypertrophic zone in the entire condyle seemed higher in the early stages but decreased in the later stages. Some apparently compacted cells in the hypertrophic zone showed proliferating cell nuclear antigen (PCNA) immunoreaction, indicating the potential for cell proliferation at the early stages. As the mice matured, in contrast, they further enlarged and assumed typical features of hypertrophic chondrocytes. Apoptotic cells were also discernible in the hypertrophic zone at the early but not later stages. Consistent with morphological configurations of hypertrophic chondrocytes, immunoreactions for alkaline phosphatase, osteopontin, and type I collagen were prominent at the later stage, but not the early stage. Cartilaginous matrices demonstrated scattered patches of mineralization at the early stage, but increased in their volume and connectivity at the later stage. Thus, the spatial and temporal occurrence of these immunoreactions as well as apoptosis likely reflect the prematurity of hypertrophying cells at the early stage, and imply a physiological relevance during the early development of the mandibular condyles. Microsc. Res. Tech. 67:325–335, 2005. © 2005 Wiley‐Liss, Inc.
Identification of mineralized elastic fibers on wet samples by SEM
Authors: Boraldi Federica , Tonelli Massimo , Gheduzzi Dealba , Ronchetti Ivonne Pasquali , Quaglino Daniela
Published: September 2005

A method is described that could be of potential use for the rapid ultrastructural identification of abnormal and fragmented elastic fibers in very small wet samples of dermal biopsies from patients affected by Pseudoxanthoma elasticum (PXE). Moreover, the method, which consists of the use of sealed capsules transparent to electrons, allows the rapid and accurate localization and detection of mineralized areas in PXE patients and of their ion composition by X‐ray microanalysis. This methodology could be of great help in any tissue disorder, especially in connective tissue disorders, characterized by structural alterations associated with ion precipitation. Microsc. Res. Tech. 67:296–299, 2005. © 2005 Wiley‐Liss, Inc.
A quantitative comparison of synaptic density following perfusion versus immersion fixation in the rat cerebral cortex
Authors: Monfils Marie‐H. , Bray Doug F. , Driscoll Ira , Kleim Jeffrey A. , Kolb Bryan
Published: September 2005

The main objective of the present study was to perform an unbiased comparison of immersion vs. perfusion techniques to assess whether we could use the former to quantify synapses through electron microscopy (EM). Using the immersion technique is ideally suited for instances in which the specimen under study could not be perfused under the standard EM protocol. Our results suggest that, despite suboptimal qualitative results, fixation by immersion allows for adequate quantification of synapses. Microsc. Res. Tech. 67:300–304, 2005. © 2005 Wiley‐Liss, Inc.
Enhanced gene expression in lung by a stabilized lipoplex using sodium chloride for complex formation
Authors: Kawakami Shigeru , Ito Yoshitaka , Fumoto Shintaro , Yamashita Fumiyoshi , Hashida Mitsuru
Published: September 2005

Background

In this study, we investigated the in vivo gene transfection efficacy of a ‘surface charge regulated’ (SCR) lipoplex, dispersed in the presence of an essential amount of NaCl during lipoplex formation.

Methods

SCR lipoplexes were prepared and their physicochemical properties were analyzed. After intravenous (i.v.) administration, transfection efficacy, distribution characteristics, and liver toxicity were evaluated in mice.

Results

At NaCl concentrations of 10 mM, the particle sizes of the SCR lipoplexes were about 120 nm and were compatible with a conventional lipoplex. However, fluorescent resonance energy transfer analysis revealed that cationic liposomes in the SCR lipoplexes increased fusion. After i.v. administration, the transfection activity in the lung of the SCR lipoplex (10 mM NaCl solution in the lipoplex) was approximately 10‐fold higher than that of the conventional lipoplex. Pharmacokinetic studies demonstrated a higher distribution in lung by the SCR lipoplex. When the gene expression levels of the SCR lipoplex and conventional lipoplex were compared, the SCR lipoplex at a dose of 30 µg was compatible with that of the conventional lipoplex at a dose of 50 µg. A significantly higher serum alanine aminotransferase (ALT) activity and TNFα concentration was observed by the conventional lipoplex (pDNA dose; 50 µg), but this was not the case for the SCR lipoplex (pDNA dose; 30 µg).

Conclusions

We demonstrated that the SCR lipoplex could enhance the transfection efficacy in the lung without increasing the liver toxicity. Hence, the information will be valuable for the future use, design, and development of lipoplexes for in vivo applications. Copyright © 2005 John Wiley & Sons, Ltd.
The cytotoxic activity of the bacteriophage λ‐holin protein reduces tumour growth rates in mammary cancer cell xenograft models
Authors: Agu Chukwuma A. , Klein Reinhard , Schwab Sonja , König‐Schuster Marielle , Kodajova Petra , Ausserlechner Michael , Binishofer Bernhard , Bläsi Udo , Salmons Brian , Günzburg Walter H. , Hohenadl Christine
Published: September 2005

Background

The potential use of gene therapy for cancer treatment is being intensively studied. One approach utilises the expression of genes encoding cytotoxic proteins. Such proteins can affect cellular viability, for example by inhibiting the translation machinery or disturbing membrane integrity. The bacteriophage Lambda (λ)‐holin protein is known to form a lesion in the cytoplasmic membrane of E. coli, triggering bacterial cell lysis and thereby enabling the release of new bacteriophage particles. The aim of this study was to evaluate whether the λ‐holin protein has a cytotoxic impact on eukaryotic cells and whether it holds potential as a new therapeutic protein for cancer gene therapy.

Methods

To explore this possibility, stably transfected human cell lines were established that harbour a tetracycline (Tet)‐inducible system for controlled expression of the λ‐holin gene. The effect of the λ‐holin protein on eukaryotic cells was studied in vitro by applying several viability assays. We also investigated the effect of λ‐holin gene expression in vivo using a human breast cancer cell tumour xenograft as well as a syngeneic mammary adenocarcinoma mouse model.

Results

The λ‐holin‐encoding gene was inducibly expressed in eukaryotic cells in vitro. Expression led to a substantial reduction of cell viability of more than 98%. In mouse models, λ‐holin‐expressing tumour cell xenografts revealed significantly reduced growth rates in comparison to xenografts not expressing the λ‐holin gene.

Conclusions

The λ‐holin protein is cytotoxic for eukaryotic cells in vitro and inhibits tumour growth in vivo suggesting potential therapeutic use in cancer gene therapy. Copyright © 2005 John Wiley & Sons, Ltd.
Analysis of drug resistance‐associated mutations in treatment‐naïve individuals infected with different genetic forms of HIV‐1 circulating in countries of the former Soviet Union
Authors: Vázquez de Parga E. , Rakhmanova A. , Pérez‐Álvarez L. , Vinogradova A. , Delgado E. , Thomson M.M. , Casado G. , Sierra M. , Muñoz M. , Carmona R. , Vega Y. , Contreras G. , Medrano L. , Osmanov S. , Nájera R.
Published: September 2005

There are few data on drug resistance‐associated mutations in the former Soviet Union since, studies have usually been focused on the env or gag genes for subtype information. This study examines the prevalence and patterns of resistance‐associated mutations to reverse transcriptase and protease inhibitors (RTI, PRI) in 278 HIV‐1‐infected treatment‐naïve subjects from countries of Eastern Europe, and defines characteristic polymorphisms of RT and PR sequences in HIV‐1 subtype A viruses. Blood samples were collected between 1997 and 2004. Plasma RNA was used for PR‐RT amplification by reverse transcription coupled with nested PCR and sequencing. Phylogenetic analysis was done with neighbor‐joining trees and bootscanning. Analysis of drug resistance mutations, with Stanford University HIV Drug Resistance Database's algorithm, resulted in an overall prevalence of 12.9% resistance to RTI and 3.9% to PRI. The most frequent substitutions in the RT region were at positions 62 and 236. V77I substitution in PR was found in 47.8% of samples. Polymorphisms in subtype A sequences were identified. This is the first study reporting the prevalence and patterns of both PRI and RTI resistance‐associated mutations in naïve HIV‐1 infected patients from the former Soviet Union. These data underline the importance of genotypic resistance testing of chronically HIV‐1‐infected patients before initiating treatment, in order to select the most suitable drug regimen. J. Med. Virol. 77:337–344, 2005. © 2005 Wiley‐Liss, Inc.
Virological and pharmacological factors associated with virological response to salvage therapy after an 8‐week of treatment interruption in a context of very advanced HIV disease (GigHAART ANRS 097)
Authors: Delaugerre Constance , Peytavin Gilles , Dominguez Stephanie , Marcelin Anne‐Genevieve , Duvivier Claudine , Gourlain Karine , Amellal Bahia , Legrand Mayeule , Raffi François , Costagliola Dominique , Katlama Christine , Calvez Vincent
Published: September 2005

Both highly potent antiretroviral drug rescue multi therapy and treatment interruption (TI) have been suggested to be effective in HIV‐1 infected‐patients with multiple treatment failure. GigHAART‐ANRS 097 was the only randomized trial during which an 8‐week TI was beneficial in heavily pre‐treated patients with multi‐drug resistant virus on resuming a multiple‐drug salvage regimen. The aim of this study was to analyze virological and pharmacological factors associated with a virological response. Clonal resistance analysis showed that although the viral population was highly mutated and nearly monoclonal at baseline, the 8‐week interruption therapy allowed the re‐emergence of more susceptible quasispecies to the subsequent salvage therapy, which were not detected by classical genotypic resistance testing. The fact that not every viral clone harbored all resistance viral mutations could explain a part of the virological response to a six to eight drug regimen for patients enrolled in the TI group. This phenomenon was associated with a transient virological response after the use of a GigHAART therapy, but was followed by the re‐emergence of baseline resistance pattern and acquisition of additional mutations in patients failing this strategy. A combined factor of protease inhibitor (PI) concentration and genotypic score, expressed as a genotypic inhibitory quotient (GIQ), was used to assess the importance of genotypic resistance and plasma drug levels in the rate of response to multiple PI combination. The GIQ of each PI used in the regimen was not associated with virological success. However, the sum of PI GIQs was predictive of a virological response. These results suggest that pharmacological enhancement might overcome viral resistance and that there is some benefit in adding the activity of several boosted‐PIs to improve the response to a salvage regimen. J. Med. Virol. 77:345–350, 2005. © 2005 Wiley‐Liss, Inc.
An outbreak of hepatitis A among homeless drug users in Rotterdam, The Netherlands
Authors: Tjon G.M.S. , Götz H. , Koek A.G. , de Zwart O. , Mertens P.L.J.M. , Coutinho R.A. , Bruisten S.M.
Published: September 2005

From the end of January to mid‐June 2004 (weeks 5–24) a hepatitis A virus (HAV) outbreak occurred among a homeless and drug user community in Rotterdam, The Netherlands. To prevent further spread of the virus within this group and to the general population, the Municipal Health Service of Rotterdam organized a mass vaccination campaign during which 83% (1,515/1,800) of the homeless people were vaccinated. As part of a national HAV typing study, blood and/or fecal samples of 30 Rotterdam HAV IgM(+) patients who fell ill during the period of 1 September 2003–1 December 2004 were tested. The tests included RT‐PCR and sequencing at the VP3‐VP1 and VP1‐P2a regions of the HAV genome. It was found that 12 homeless people, one family member of a homeless person and two people without a known risk were infected with a unique subtype 3a strain. Four of the homeless patients became ill after vaccination and were probably infected at the time. This study shows that Dutch homeless people and drug users involved in HAV outbreaks should be offered HAV vaccine actively to prevent further spread of the infection. Furthermore, it was shown by molecular techniques that the unique subtype 3a strain was not found before the Rotterdam outbreak or afterwards, indicating that the mass vaccination campaign was successful. J. Med. Virol. 77:360–366, 2005. © 2005 Wiley‐Liss, Inc.
Stopping lamivudine therapy after biochemical breakthrough may be a feasible option in selected HBeAg‐positive patients
Authors: Jang Jeong Won , Choi Jong Young , Bae Si Hyun , Yoon Seung Kew , Kim Chang Wook , Nam Soon Woo , Lee Chang Don , Lee Young Sok , Cha Sang Bok , Chung Kyu Won
Published: September 2005

Drug resistance remains a major problem during lamivudine treatment for patients with hepatitis B virus infection. Continuing or stopping lamivudine in patients with biochemical breakthrough is an issue that has not been investigated extensively. Particularly, HBeAg seroconversion and its durability achieved in these settings have never been studied so far. In this study, we compared the clinical outcomes including HBeAg seroconversion in hepatitis B e antigen (HBeAg)‐positive patients who either continued or stopped lamivudine treatment after biochemical breakthrough. This was a retrospective cohort study of 73 consecutive patients with biochemical breakthrough followed up for at least 12 months. Of these, 35 (Group I) stopped and the remaining 38 (Group II) continued lamivudine therapy after breakthrough. During 12 months, there was a tendency towards higher rates of HBeAg seroconversion in Group I compared with Group II (P = 0.078). By multivariate analysis, age ≥45 years and acute exacerbation within 6 months of stopping lamivudine were the two independent predictors of HBeAg seroconversion for Group I (P = 0.034 and P = 0.012, respectively). There was no difference in the overall incidence of acute exacerbation and hepatic decompensation between the two groups (P = 0.333 and P = 0.555, respectively). Six patients (17.1%) in Group I developed hepatic decompensation, for which bilirubin ≥1.2 mg/dl was an independent predictor (P = 0.002). The therapeutic gain in continuing lamivudine for biochemical breakthrough is questionable. This study suggests that discontinuation of lamivudine may be an option in older patients with normal bilirubin level. J. Med. Virol. 77:367–373, 2005. © 2005 Wiley‐Liss, Inc.
Hepatitis B virus X protein upregulates survivin expression in hepatoma tissues
Authors: Zhang Xiaodong , Dong Nan , Yin Lihui , Cai Na , Ma Hongtao , You Jiacong , Zhang Hang , Wang Honghui , He Ran , Ye Lihong
Published: September 2005

The hepatitis B virus X protein (HBx) plays an important role in the development of hepatocellular carcinoma (HCC). The relationship was examined between HBV antigens and IAP (inhibitor of apoptosis) family in development of HCC. The expression levels of HBV antigens (HBsAg, HBcAg, and HBxAg) and members of the IAP family (survivin, XIAP, cIAP‐1, and cIAP‐2) were detected immunohistochemically in tissues from 34 cases of HCC and 30 cases of liver cirrhosis. The positive rate of survivin was higher than these three molecules in all three tissue types (P < 0.05). The positive rates of HBxAg and survivin were high in HCC (76.5% and 88.2%), paratumor (85.3% and 91.2%), and liver cirrhosis (100% and 93.3%) tissues, with no significant differences between the survivin‐ and HBxAg‐positive rates (each P > 0.05). To examine the effect of HBx on survivin expression, plasmid pCMV‐X (encoding the HBx gene) was transfected transiently with or without plasmid pcDNA3‐sur (encoding the survivin gene) into H7402 hepatoma cells and L‐O2 human normal liver cells. Cells over‐expressing HBx alone showed increased apoptosis along with a dose‐dependent increase in survivin levels. However, co‐expression of survivin inhibited the HBx‐induced apoptosis. To examine the effect of HBx on survivin in hepatoma cells without apoptosis, plasmid pCMV‐X was transfected stably into human hepatoma H7402 cells and L‐O2 cells. These H7402‐X and L‐O2‐X cells showed high‐level expression of both HBx and survivin, but did not show apoptosis. The addition of pSilencer 3.0‐X, an RNAi vector targeting the HBx gene, reduced the expression levels of survivin protein in H7402‐X cells. Collectively, these data demonstrate that HBx upregulates survivin expression in hepatoma tissues, suggesting that HBx and survivin may both be involved in carcinogenesis of HCC. J. Med. Virol. 77:374–381, 2005. © 2005 Wiley‐Liss, Inc.
Molecular epidemiology of the hepatitis C virus in Western Siberia
Authors: Shustov A.V. , Kochneva G.V. , Sivolobova G.F. , Grazhdantseva A.A. , Gavrilova I.V. , Akinfeeva L.A. , Rakova I.G. , Aleshina M.V. , Bukin V.N. , Orlovsky V.G. , Bespalov V.S. , Robertson B.H. , Netesov S.V.
Published: September 2005

Western Siberia is the region with little information on the prevalence of hepatitis C virus (HCV) infection, genotypic diversity of HCV isolates and risk factors. A molecular epidemiological survey was conducted to clarify these issues. Four groups of volunteers were included in a cross‐sectional study (n = 500 in each group): health care workers; daycare patients from a hospital for drug users, daycare patients from an AIDS prevention and control center; and persons admitted to a local general practice clinic for any reason (outpatients). The anti‐HCV IgG prevalence was 4.6% in health care workers, 48.0% in a narcological center, 35.8% in AIDS center, and 5.6% in outpatients. HCV RNA was found in 79.3%–86.3% of seropositives. A total of 388 HCV isolates were genotyped by direct sequencing and phylogenetic analysis of the 5′‐UTR and NS5B regions of HCV genome. The genotypes distribution was: 1b—50.3%, 2a—4.4%, 2c—0.3%, 3a—44.8%. One isolate (0.3%) could not be typed unambiguously. This genotypic diversity is intermediate between that of European Russia and China. Genotype 1 prevailed in an older age group (75% among 51–60 years old), and genotype 3 was most prevalent in young people (51.4% in 16–20 years old). A statistically significant (P < 0.05) increase in risk was found in intravenous drug users (odds ratio (OR) = 77.5), unemployed persons (OR = 16.3), persons having >4 sexual partners during lifetime (OR = 4.3), and male homosexuals (OR = 6.6). J. Med. Virol. 77:382–389, 2005. © 2005 Wiley‐Liss, Inc.
Distribution and heterogeneity of hepatitis C genotypes in hepatitis patients in Cameroon
Authors: Pasquier Christophe , Njouom Richard , Ayouba Ahidjo , Dubois Martine , Sartre Michèle Tagni , Vessière Aurelia , Timba Isabelle , Thonnon Jocelyn , Izopet Jacques , Nerrienet Eric
Published: September 2005

Hepatitis C virus infects humans world‐wide. The virus genome varies greatly and it has several genotypes. HCV infection is highly prevalent in Central Africa and Cameroon. Initial studies on the genetic variability of HCV showed infection with HCV genotypes 1, 2, and 4. We have now sequenced the NS5b and E2 regions of 156 HCV isolates collected from patients presenting for diagnosis in Yaounde and used the data to describe the distribution of HCV genotypes and subtypes in patients with hepatitis in Cameroon. Genotype 1 was more frequent than Genotypes 4 and 2. Genotypes 1 and 4 were highly heterogeneous, containing many subtypes described previously (1b, 1c, 1e, 1h, 1l, 4f, 4t, 4p, 4k) and unsubtyped groups. There was a systematic phylogenetic concordance between NS5b and E2 sequence clustering. The Genotype 2 sequences did not vary. Neither subject age nor gender influenced HCV distribution. HCV Genotypes 1 and 4 are very heterogeneous in Cameroon, perhaps due to ancient infections. The homogeneity of HCV Genotype 2 indicates its more recent introduction from western Africa. J. Med. Virol. 77:390–398, 2005. © 2005 Wiley‐Liss, Inc.
Identification of novel HCV subgenome replicating persistently in chronic active hepatitis C patients
Authors: Yagi Shintaro , Mori Kenich , Tanaka Eiji , Matsumoto Akihiro , Sunaga Fumiko , Kiyosawa Kendo , Yamaguchi Kenjiro
Published: September 2005

In an effort to clarify the life cycle of HCV, the HCV genome in liver biopsies taken from chronic active hepatitis C patients undergoing interferon treatment was investigated. Molecular cloning by long distance reverse‐transcription polymerase chain reaction (RT‐PCR) revealed that the HCV genome in two patients with high viral loads in the liver had in‐frame deletions of approximately 2 kb between E1 and NS2, which encode the E1–NS2 fusion protein and six other HCV proteins: core, NS3, NS4A, NS4B, NS5A, and NS5B. Among the remaining 21 chronic active hepatitis C patients, these types of deletion were found in another two patients and in two hepatocellular carcinoma patients. Out‐of‐frame deletions in the structural region were isolated from the other five patients, but the dominant RT‐PCR products were non‐truncated genomes. Retrospective analysis of a series of serum samples taken from a patient carrying the subgenome with the in‐frame deletion revealed that both the subgenome and the full genome persisted through the 2‐year period of investigation, with the subgenome being predominant during this period. Sequence analysis of the isolated cDNA suggested that both the subgenome and the full genome evolved independently. Western blotting analysis of HCV proteins from the HCV subgenome indicated that they were processed in the same way as those from the full genome. HCV subgenomes thus appear to be involved in the HCV life cycle. J. Med. Virol. 77:399–413, 2005. © 2005 Wiley‐Liss, Inc.
Analysis of cytomegalovirus DNA polymerase (UL54) mutations in solid organ transplant patients receiving valganciclovir or ganciclovir prophylaxis
Authors: Boivin Guy , Goyette Nathalie , Gilbert Christian , Covington Emma
Published: September 2005

We previously reported the absence of CMV UL97 (kinase) gene resistance mutations up to 12 months post‐transplant following 100 days of valganciclovir prophylaxis, and a low incidence of resistance mutations following 100 days of oral ganciclovir prophylaxis in a prospective multicenter study in solid organ transplant recipients excluding lung transplants. Herein, we report UL54 (DNA polymerase) gene sequencing results for all patients with previous UL97 PCR‐positive samples (n = 99) in our study. One UL54 resistance mutation (L545S known to confer ganciclovir and cidofovir resistance) was detected in a routine day‐100 sample from an asymptomatic patient who received oral ganciclovir. Notably, this CMV UL54 mutation occurred in the absence of a UL97 mutation. Additionally, new UL54 variants were observed. Thus, emergence of CMV UL54 mutations in the absence of UL97 mutations is a rare but possible event that is not necessarily associated with detrimental clinical outcome in solid organ transplant recipients. J. Med. Virol. 77:425–429, 2005. © 2005 Wiley‐Liss, Inc.
Characterization of the homo‐ and heterotypic immune responses after natural norovirus infection
Authors: Rockx Barry , Baric Ralph S. , de Grijs Ingrid , Duizer Erwin , Koopmans Marion P.G.
Published: September 2005

Noroviruses (NoV) are a genetically and antigenically diverse group of viruses that are common causes of outbreaks of gastroenteritis in humans of all ages. Limited information has been obtained on type specificity of the NoV immune response. In this study, we characterized the homologous and heterologous antibody responses in adults from 13 outbreaks, representing 4 different NoV genotypes. NoV specific IgG and IgA antibodies were determined as well as the increase of antibody avidity. In addition, antibody‐mediated blocking of NoV binding to its putative receptor was evaluated. Both homologous and heterologous serological responses were detected after NoV infection. The avidity of antibodies could not be used to distinguish between homologous and heterologous antibody responses. However, a homologous blocking response but not a heterologous response was detected after infection with NoV belonging to genogroup II.4 by a NoV ligand binding inhibition assay. Infection with NoV induces antibodies that can block virus ligand interactions. In contrast with all currently known antibody detection assays for NoV, this can be used as a type specific assay and may be an alternative for studying neutralizing antibodies. J. Med. Virol. 77:439–446, 2005. © 2005 Wiley‐Liss, Inc.
A novel method for measuring distribution of orientation of one‐dimensional ZnO using resonance Raman spectroscopy
Authors: Zhang Xinghua , Liu Yichun , Chen Shihao
Published: September 2005

A spectroscopic method of characterizing the growth orientation of low‐dimensional ZnO structures is proposed on the basis of the angular dispersion of polar phonon modes. According to this approach, the orientational distribution of a randomly oriented one‐dimensional ZnO structure can be measured quantitatively by studying the line‐shape of polar phonon bands. Multiphonon resonance Raman spectroscopy was used to investigate the details of the polar optical phonon mode. Copyright © 2005 John Wiley & Sons, Ltd.
Raman spectroscopy of beaverite and plumbojarosite
Authors: Frost Ray L. , Wills Rachael‐Anne , Martens Wayde
Published: September 2005

Raman spectroscopy has been used to characterise the compound of formula Pb(Fe3+, Cu2+)3(SO4)2(OH)6, equivalent to synthetic beaverite, one of the jarosite subgroup minerals. The mineral is characterised by multiple OH stretching vibrations attributed to non‐equivalent OH units in the structure. Multiple vibrations are observed for the SO42− stretching vibrations indicating the non‐equivalence of the sulphate units in the structure. This multiplicity is also reflected in the ν2 and ν4 bending modes. The Raman spectrum of beaverite is significantly different from that of plumbojarosite, for which multiple bands are not observed. Copyright © 2005 John Wiley & Sons, Ltd.
Raman spectroscopy shows antifreeze glycoproteins interact with highly oriented pyrolytic graphite
Authors: Cui Y. , Turner G. , Roy U. N. , Guo M. , Pan Z. , Morgan S. , Burger A. , Yeh Yin
Published: September 2005

Raman spectra of a mixture of antifreeze glycoproteins (AFGP) 6, 7 and 8 adsorbed on a highly oriented pyrolytic graphite (HOPG) surface spanning the range of 50 to 4500 cm−1 were measured. It was found that the Raman signature of AFGP had been significantly altered because of its interaction with the HOPG surface; when compared with the spectra of bulk AFGP compounds, at least three new peaks were observed around 860, 940 and 1000 cm−1, and two other peaks that were not well resolved in the bulk spectra around 1050 and 1155 cm−1 were enhanced and well discriminated. The peak positions depended on the facet of HOPG presented to the proteins, and their relative intensities depended on the concentration of AFGP used in the adsorption experiment. The results indicated the possibility of multiple binding sites of AFGP on HOPG surfaces. Copyright © 2005 John Wiley & Sons, Ltd.
Potential use of visible and near infrared reflectance spectroscopy for the estimation of nitrogen fractions in forages harvested from permanent meadows
Authors: Valdés Carmen , Andrés Sonia , Giráldez F Javier , García Ricardo , Calleja Alfredo
Published: September 2005

One‐hundred and ninety‐two herbage samples from permanent meadows located in the mountains of León (NW Spain) were analyzed for total nitrogen (total N), nitrogen in trichloroacetic acid precipitated matter (TCAN), borate–phosphate buffer insoluble nitrogen (BPBN), neutral‐detergent insoluble nitrogen (NDIN) and acid‐detergent insoluble nitrogen (ADIN). These data were used to calculate the partition of nitrogen fractions proposed by the Cornell Net Carbohydrate and Protein System (CNCPS): A (total N − TCAN), B1 (TCAN − BPBN), B2 (BPBN − NDIN), B3 (NDIN − ADIN) and C (ADIN). Near‐infrared reflectance spectroscopy (NIRS) technology accurately predicted total N, TCAN and NDIN, as judged by coefficient of determination (R2) and ratio performance deviation (RPD) values greater than 0.90 and 2.5, respectively. The rest of the insoluble N fractions (BPBN and ADIN) were predicted with less accuracy by NIRS. Estimations of CNCPS N fractions (A, B1, B2, B3 and C) using visible–NIR spectra of forage samples did not allow accurate predictions (R2 < 0.90; RPD < 2.5). Copyright © 2005 Society of Chemical Industry
Consumer evaluation of novel kiwifruit: willingness‐to‐pay
Authors: Jaeger Sara R , Harker F Roger
Published: September 2005

This paper introduces a research methodology that adds sophistication to the innovation process for agricultural goods. In recognition of the importance of innovation for the long‐term success of firms and the need for market analysis that goes beyond traditional sensory and consumer preference testing, it is demonstrated how to measure the monetary value consumers place on new market goods. In experimental markets, New Zealand (NZ) consumers were willing to pay on average approximately NZ$0.44 per 100 g to exchange the common green‐fleshed Hayward variety for a new‐to‐market red‐ and yellow‐fleshed kiwifruit variety. At the time of the study this was equivalent to a 179% retail price premium. Knowledge that this new variety was not genetically modified increased the price premium to 240%. These results were elicited from convenience samples and should be treated with caution if used as the sole basis for price setting. Copyright © 2005 Society of Chemical Industry
Farmers' knowledge and perceptions of soil erosion and conservation measures in the Central Highlands, Kenya
Authors: Okoba B. O. , De Graaff J.
Published: September 2005

A lack of appreciation of Kenyan farmers' knowledge and their perceptions of soil erosion and soil conservation measures was the reason for low adoption of recommended technologies. This research was carried out to identify the criteria that farmers used to distinguish farm‐types and to use these types to evaluate different knowledge levels and perceptions of soil erosion and existing soil and water conservation (SWC) measures in the Central Highlands of Kenya. Community meetings and semi‐structured household surveys were carried out in a small catchment, with 120 households. Results partly support the idea of using farmer‐developed criteria to distinguish among land managers with reference to a farm‐type classification. Criteria distinguishing three classes of land managers (good, moderate and poor) were significant with regard to the following land husbandry practices: use of hybrid or recycled seed and use of organic and/or inorganic fertilizers. Farmers were aware of on‐going soil erosion and of several erosion control measures. Whereas a majority of farmers preferred grass‐strips as a SWC measures, they did not recognize agroforestry as a form of SWC measure. Farmers perceived that SWC measures could successfully increase crop yields and soil‐water retention, and increase land value. In general, farmers did not perceive that SWC measures successfully prevented erosion phenomena, given the evidence of on‐site erosion indicators. They attributed the continued erosion to high rainfall, steep slopes, lack of maintenance and poorly designed SWC measures. They did not consider poor soil‐cover, up‐down tillage and tall trees to be the causes of erosion. Farmers faced several constraints in adopting SWC measures: lack of labour, tools, capital and know‐how to construct the measures. Copyright © 2005 John Wiley & Sons, Ltd.