bluuebunny/crossref_metadata_2025_split huggingface dataset:
Types of vascular wall as related to vasa vasorum in common tree shrew (Tupaia glis)
Authors: Chuncher Sununta , Somana Reon 
Published: September 2005

AbstractBlood 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

AbstractMicromass 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

AbstractThe 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

AbstractA 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

AbstractThe 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

AbstractBackgroundIn 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.MethodsSCR 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.ResultsAt 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).ConclusionsWe 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

AbstractBackgroundThe 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.MethodsTo 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.ResultsThe 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.ConclusionsThe 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

AbstractThere 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

AbstractBoth 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

AbstractFrom 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

AbstractDrug 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

AbstractThe 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

AbstractWestern 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.