bluuebunny/crossref_metadata_2025 huggingface dataset:
Morphological alteration and biological properties of hepatocytes not related to tumorigenesis following transfection with HCV core protein*
Authors: Gu J. , Wang L. , Che Y. , Liu L. , Jiang L. , Dong S. , Li W. , Li Q.
Published: January 2005

Summary.  The hepatitis C virus (HCV) core protein is supposed to play a critical role in HCV‐mediated human liver disease with its capabilities to regulate the growth rate of hepatocytes and to partially contribute to the pathogenesis of hepatocellular carcinoma in association with cellular oncogenes. In this study, to analyse the possible pathological mechanism of the HCV core protein, human primary embryo hepatocytes transfected with HCV core were monitored by immunofluorescence, reverse transcriptase‐polymerase chain reaction (RT‐PCR) and Western blot. The morphological changes and biological properties of the transfected hepatocytes were also studied. The results showed that the HCV core gene integrated in the cellular genome and the protein expressed in the transfected hepatocyte, could be detected following serial passage at both the mRNA and protein level. The proliferation assays indicated that hepatocytes transfected with the HCV core gene alone did not exhibit any tumorigenic tendency. Meanwhile, the morphological alterations of these cells demonstrated obvious changes in size, and large vacuolar degeneration. In conclusion, the hepatocytes transfected with the HCV core gene revealed that the core protein expressed induced pathological changes of degeneration, probably related indirectly to tumorigenicity.
Gene expression profile of T‐cell‐specific chemokines in human hepatocyte‐derived cells: evidence for a synergistic inducer effect of cytokines and hepatitis C virus proteins
Authors: Apolinario A. , Majano P. L. , Lorente R. , Núñez O. , Clemente G. , García‐Monzón C.
Published: January 2005

Summary.  Increased levels of chemokines (CK) in chronic hepatitis C virus (HCV) infection have been found. Given that NS5A and core can function as transcriptional transactivators, we aimed to determine whether these HCV proteins might induce CK gene expression in human hepatocyte‐derived cells. We assessed (i) regulated upon activation, normal T cells expressed and activated (RANTES), interferon γ‐inducible protein‐10 (IP‐10), and monokine induced by interferon‐γ (MIG) mRNA levels in NS5A and core stably transfected Chang liver (CHL) cells, stimulated or not with a cytokine mixture (CM), by reverse transcriptase‐polymerase chain reaction (RT‐PCR) and (ii) quantitative enzyme‐linked immunosorbent assay (ELISA) measurements of these CK in the supernatants of CHL cells. Induction of RANTES transcripts in resting HCV‐transfected cells was clearly observed, being augmented fourfold in resting NS5A‐transfected cells and threefold in resting core‐transfected cells over that in resting mock‐transfected (control) cells, as well as to a similar extent in CM‐stimulated NS5A‐ and core‐transfected cells. Increased RANTES secretion followed the same pattern observed for mRNA expression. Both IP‐10 and MIG, such as mRNA and protein levels, were undetectable in resting HCV‐transfected and ‐untransfected cells, whereas IP‐10 and MIG mRNA expression was increased by seven‐ and fivefold in CM‐stimulated NS5A‐transfected cells and by 10‐ and 3.5‐fold in CM‐stimulated core‐transfected cells, respectively, above that in CM‐stimulated control cells. IP‐10 and MIG secretion was enhanced by 2.6‐ and threefold in CM‐stimulated NS5A‐transfected cells and by 3.6‐fold and 3.7‐fold in CM‐stimulated core‐transfected cells, respectively over that in CM‐stimulated control cells. These results demonstrate that NS5A and core proteins, alone or by the synergistic effect of cytokines, are capable of upregulating RANTES, IP‐10 and MIG gene expression in cultured human hepatocyte‐derived cells.
Activation and function of hepatic NK cells in hepatitis B infection: an underinvestigated innate immune response
Authors: Chen Y. , Wei H. , Gao B. , Hu Z. , Zheng S. , Tian Z.
Published: January 2005

Summary.  Natural killer (NK) cells are abundant in the normal liver, accounting for around one‐third of intrahepatic lymphocytes and are important in the defence against hepatitis B virus (HBV) infection as innate immune responses. In this review, we discuss the mechanisms of hepatic NK cell activity against HBV. Whether directly activated by HBV infection or indirectly activated by other lymphocytes such as NKT cells or antigen‐presenting cells (APCs), hepatic NK cells exert their anti‐viral functions by natural cytotoxicity and production of high levels of cytokines. However, activated NK cells play an important role in regulating adaptive immune responses by interaction with other lymphocytes such as T, B and APCs. In addition, NK cells may contribute to the lymphocyte‐mediated liver injury during HBV infection that was previously considered to be mediated only by CD8+ T cells or/and NKT cells.
The minimum number of clones necessary to sequence in order to obtain the maximum information about hepatitis C virus quasispecies: a comparison of subjects with and without liver cancer
Authors: Gao G. , Stuver S. O. , Okayama A. , Tsubouchi H. , Mueller N. E. , Tabor E.
Published: January 2005

Summary.  Most studies of hepatitis C virus (HCV) quasispecies have reported the results of sequencing only three to five clones per sample. The possibility that sequencing so few clones might not provide a representative picture of the quasispecies present in a sample has never been evaluated. The present study was conducted to evaluate whether sequencing greater numbers of clones results in better information about the HCV quasispecies number and distribution, and to compare the HCV quasispecies in liver cancer cases and controls. RNA was extracted from serial serum samples from six subjects with HCV‐associated liver cancer and 11 age‐ and sex‐matched HCV‐infected controls without liver cancer. The hypervariable region 1 (HVR1) of the HCV genome was amplified, cloned, and sequenced. For further studies of 12 serum samples from two liver cancer cases and two matched controls, successive groups of 10 additional clones were sequenced up to a total of 50 clones per serum sample. When only 10 clones were sequenced from each specimen, no consistent differences were seen between the number of HCV quasispecies in the six liver cancer cases and the 11 controls. However, sequencing 40 clones from each of 12 samples from two liver cancer cases and two controls revealed a greater number of quasispecies in liver cancer cases than in controls. Testing an additional 10 clones (50 clones per sample) did not significantly increase the number of quasispecies detected.
The effect of adherence to therapy on sustained response in daily or three times a week interferon alpha‐2b plus ribavirin treatment of naïve and nonresponder chronic hepatitis C patients
Authors: Raptopoulou M. , Tsantoulas D. , Vafiadi I. , Ketikoglou I. , Paraskevas E. , Vassiliadis T. , Kanatakis S. , Hatzis G. , Sidiropoulos L. , Akriviadis E.
Published: January 2005

Summary.  The aim was to demonstrate adherence to treatment has been suggested to enhance rates of sustained response in patients with hepatitis C. In this study, we evaluated the effect of drug dosage reduction or the duration of the expected therapy in patients treated with interferon (IFN)‐alpha2b plus ribavirin. Virologic response rates were re‐analysed according to compliance to therapy in (i) 301 naïve and (ii) 142 nonresponders to previous IFN therapy treated with either IFN 5 MU TIW for 8 weeks followed by IFN 3 MU TIW for 40 weeks plus ribavirin or IFN 3 MU QD for 16 weeks followed by IFN 3 MU TIW for 24 weeks plus ribavirin. Patients were separated into those who adhered to ≥80% of their intended treatment schedule (dose of both drugs and duration) and those who did not. Compliance to treatment resulted in significantly higher response rates in both groups of patients: 43.93% compared with 6.90% of noncompliant naïve patients and 30.77% compared with 10.53% of nonresponder patients. Compliance to treatment was found to have a similar effect when the results were analysed according to HCV genotype. Our findings suggest that compliance to treatment for ≥80% of the intended treatment schedule results in significantly higher sustained response rates in both naïve and nonresponder patients. Consequently, every effort should be made to improve patient adherence to therapy.
Seroepidemiology of hepatitis A in patients with chronic liver disease
Authors: Saab S. , Lee C. , Shpaner A. , Ibrahim A. B.
Published: January 2005

Summary.  Hepatitis A virus (HAV) rarely causes fulminant hepatic failure in the general population. Yet it is a cause of significant morbidity and mortality in patients with chronic liver disease (CLD), in whom routine HAV vaccination is recommended. However, studies of HAV seroprevalence and exposure predictors in populations with CLD are scarce. We have studied a cohort of 473 patients with various causes of CLD between July 2000 and June 2002. Patients were stratified on the basis of age, gender, ethnicity and aetiology of liver disease. The HAV seroprevalence in patients with CLD was compared with that in the general population. We used a logistic regression analysis to identify independent predictors of HAV exposure. Of the 473 patients studied, HAV seroprevalence was available for 454 individuals. HCV, HBV, alcohol, and HCV and alcohol were the causes of CLD in 337, 72, 37 and eight patients, respectively. The overall HAV seroprevalence was 55% in the studied cohort. The age‐stratified HAV prevalence for ages 21–30, 31–40, 41–50, 51–60, 61–70 and greater than 70 years was 44, 51, 44, 63, 65 and 64%, respectively. Hispanic ethnicity, Asian ethnicity, alcohol use and ages of 51–70 years were found to be independent predictive variables of prior exposure to HAV. HAV infection in patients with CLD causes considerable morbidity and mortality. We demonstrated that age‐stratified seroprevalence of HAV in patients with CLD of various aetiologies is significantly higher than that of the general population, and identified several independent predictors of HAV prior exposure.
Patient concerns regarding chronic hepatitis C infections
Authors: Minuk G. Y. , Gutkin A. , Wong S. G. , Kaita K. D. E.
Published: January 2005

Summary.  Counselling of patients with chronic hepatitis C infections is often limited to discussions regarding how the virus is transmitted and what can be done to decrease the risk of transmission to others. The purpose of the present study was to document the principal concerns of newly diagnosed and follow‐up patients with chronic hepatitis C, and thereby enhance counselling strategies and content. Seventy newly diagnosed and 115 follow‐up patients with chronic hepatitis C virus (HCV) infection were initially asked in an open‐ended manner (volunteered concerns) and then to prioritize from a prepared list of seven potential concerns (prioritized concerns), to identify those concerns that were of utmost importance to them. The most common volunteered concerns of newly diagnosed patients in decreasing order were: disease progression (27%), premature death (19%), infecting family members (13%), side‐effects of treatment (11%) and miscellaneous others. In decreasing order, prioritized concerns included: infecting family members, development of liver cancer, infecting others, development of cirrhosis, social stigma of having liver disease, need for liver transplant and loss of employment. The principal volunteered and prioritized concerns of follow‐up patients were similar to those of newly diagnosed patients. Volunteered and prioritized concerns were relatively consistent across the different genders, age groups, ethnic backgrounds, education level, marital status, employment, modes of viral acquisition and in the case of follow‐up patients, duration of follow‐up. These results indicate that health care providers who focus counselling efforts exclusively on viral transmission are unlikely to address other important concerns of newly diagnosed and follow‐up patients with chronic HCV infection.
Prophylactic SSRI during interferon alpha re‐therapy in patients with chronic hepatitis C and a history of interferon‐induced depression
Authors: Kraus M. R. , Schäfer A. , Al‐Taie O. , Scheurlen M.
Published: January 2005

Summary.  Only limited data are available on selective serotonin re‐uptake inhibitor (SSRI) prophylaxis for antiviral re‐treatment in hepatitis C patients with previous interferon‐induced major depressive episodes. Therefore, we investigated the efficacy and safety of secondary SSRI prophylaxis in these patients. In a prospective and longitudinal study, repeated psychometric testing (Hospital Anxiety and Depression Scale) was performed before, during, and after antiviral re‐treatment. Chronic hepatitis C virus (HCV)‐infected patients, who had been psychometrically monitored during an unsuccessful previous antiviral therapy, and had developed major depression were included. Interferon re‐therapy with SSRI prophylaxis was started (n = 8). The reference group was comprised of HCV patients without a history of interferon‐associated depression and also a group who were previously unsuccessfully treated with interferon and were re‐treated without SSRI prophylaxis (n = 9). All patients receiving SSRI prophylaxis were able to complete interferon re‐therapy as scheduled. As in the first therapeutic course, depression scores were significantly elevated during re‐treatment also (P < 0.001). Depression scores were significantly lower (P =0.036) during interferon re‐therapy with SSRI prophylaxis. Reference group subjects showed similar depression scores during first therapy and re‐therapy (P > 0.05). In conclusion, hepatitis C patients with a history of interferon‐induced major depression can be successfully re‐treated with peginterferon/ribavirin and concomitant SSRI prophylaxis. In these patients, SSRI prophylaxis is safe and efficacious and should be considered, if antiviral re‐therapy is indicated.
Therapeutic modalities in hepatitis C: challenges and development
Authors: Moreno‐Otero R.
Published: January 2005

Summary. Our understanding of the pathogenicity of hepatitis C virus (HCV) is based on patients infected chronically for >20 years. The lack of a suitable animal model, the narrow host range of the virus, and the protracted onset of liver disease induced by HCV have hampered advances in treatment. In spite of these problems, we identified patient and viral characteristics that predict responses to current therapies, including HCV genotype, viral load, body weight, age, liver histology, co‐infection with HIV and treatment adherence and tolerance. Interferon (IFN) alpha was the first therapy for chronic HCV infection. The combination of IFN plus ribavirin increases sustained virological response rates compared with IFN alone. Two pegylated IFNs have been developed and are widely approved for the treatment of chronic hepatitis C: peginterferon alpha‐2a (40 KD), and pegylated IFN alpha‐2b (12 KD). These products have reduced systemic clearance, prolonged half‐lives and reduced antigenicity compared with conventional IFN. The reduced clearance results in sustained plasma levels of the drug and allows for once‐weekly dosing. Pegylated IFN alpha‐2b (12 KD) has a small, linear polyethylene glycol (PEG) moiety and has an intermediate duration of activity; peginterferon alpha‐2a (40 KD) incorporates a large, branched‐chain PEG moiety and has a longer half‐life than both conventional IFN alpha and pegylated IFN alpha‐2b (12 KD). The combination of a pegylated IFN plus ribavirin significantly increases sustained virological response rates compared with conventional IFN plus ribavirin in patients with chronic hepatitis C and is now recognized as the standard of care for these patients.
Predicting progressive hepatic fibrosis stage on subsequent liver biopsy in chronic hepatitis C virus infection1
Authors: Collier J. D. , Woodall T. , Wight D. G. D. , Shore S. , Gimson A. E. , Alexander G. J. M.
Published: January 2005

Summary.  Retrospective cross‐sectional studies indicate that 20% with chronic hepatitis C virus (HCV) infection become cirrhotic within 20 years. Known risk factors for advanced hepatic fibrosis include age at time of infection, male sex, excess alcohol consumption and cytokine polymorphisms. Prospective study to assess and identify factors predictive of change in hepatic fibrosis stage in chronic HCV infection by interval protocol liver biopsy was performed. One hundred and five patients with paired liver biopsy specimens separated by a mean 41 months were recruited from a cohort of 823 HCV carriers. Five per cent developed worsening hepatic fibrosis by more than two stages. In 43% there was no change in fibrosis stage. Excessive alcohol intake currently (P = 0.037) or previously (P = 0.07) predicted progression. In contrast, always having a normal alanine transaminase (P = 0.038) and always being negative in serum for HCV RNA (P =0.067) predicted no progression. Three models were developed to predict outcome. Progressive fibrosis was predicted by baseline fibrosis (P = 0.018), steatosis (P = 0.02) and age (P = 0.017). The rate of progressive fibrosis was predicted by baseline fibrosis (P = 0.0002), steatosis (P =0.039) and lobular inflammation (P = 0.09). Fibrosis stage on the second biopsy was predicted by baseline fibrosis alone (P = 0.01). The rate of progression varies widely. Alcohol misuse is an important co‐factor. Progressive fibrosis can be predicted at first liver biopsy, where baseline fibrosis is most critical, allowing targeted therapy for those with early disease and a significant risk of progression.
Treatment of hepatitis D
Authors: Niro G. A. , Rosina F. , Rizzetto M.
Published: January 2005

Summary. Delta virus related chronic hepatitis is difficult to treat. The response toα‐interferon (IFN), which still represents the only therapy for chronic hepatitis D, varies widely and occurs at different times from the beginning of treatment. The rate of response is proportional to the dose of IFN, with 9 million units (MU) three times a week being more effective than 3 MU thrice weekly. Sustained responses are unusual and are accompanied by the clearance of serum hepatitis B virus surface antigen (HBsAg), seroconversion to anti‐HBs and improvement of liver histology. Although disease of a short‐standing may respond better to therapy, clear predictors of response are still unidentified. Besides IFN, other therapeutic approaches such as immunosuppressive drugs, acyclovir, ribavirin and thymosin, have been unhelpful. Available evidence does not support the use of deoxynucleotide analogues. Famciclovir has no effect on disease activity and hepatitis D virus (HDV)‐RNA levels. Twelve‐ or 24‐month lamivudine treatment does not significantly affect biochemical, virological or histological parameters. Pegylated‐IFN could represent a reasonable therapeutic option in the long‐term treatment required for chronic hepatitis D. Antisense oligonucleotides and prenylation inhibitors hold promise as therapeutic agents of the future. Liver transplantation provides a valid option for end‐stage HDV liver disease; the risk of re‐infection is lower for HDV than for HBV under long‐term administration of hyperimmune serum against HBsAg. Molecularly tailored drugs capable of interfering with crucial viral replicative processes of HDV appear to be the best prospect in the treatment of hepatitis D.
Effects of selective dopamine receptor subtype agonists on cardiac contractility and regional haemodynamics in rats
Authors: Polakowski James S , Segreti Jason A , Cox Bryan F , Hsieh Gin C , Kolasa Teodozyj , Moreland Robert B , Brioni Jorge D
Published: January 2005

1. Activation of dopamine (DA) receptors produces cardiovascular responses such as vasodilation and hypotension. However, knowledge of the role of specific dopamine receptor subtypes (especially D3 and D4) in the cardiovascular system is limited. The objective of the present study was to characterize the haemodynamic and cardiac responses to agonists with selectivity for D1, D2, D3 and D4 receptor subtypes.

2. Inactin‐anaesthetized rats were instrumented to measure regional haemodynamic and cardiac contractility responses with slow intravenous infusion of agonists.

3. Fenoldopam (a D1 receptor agonist) decreased (P < 0.05) renal vascular resistance beginning at a dose of 3 µmol/kg. Infusion of PNU‐95666E (a D2 receptor agonist) produced dose‐dependent decreases (P < 0.05) in mean arterial pressure (MAP), heart rate (HR) and hindquarter vascular resistance (HQVR). Administration of BP897 (a partial D3 receptor agonist) decreased (P < 0.05) MAP and HQVR at 3 µmol/kg. PD168077 (a D4 receptor agonist) caused significant increases in HQVR at 1 µmol/kg. None of the compounds tested elicited significant changes in cardiac contractility.

4. Using selective agonists of dopamine receptor subtypes, the present studies characterize distinct cardiovascular effects in anaesthetized rats. Consistent with its well‐defined effects as a D1 receptor agonist, fenoldopam administration resulted in renal vasodilation. Similar to earlier studies using the non‐selective D2‐like receptor agonist quinpirole, selective agonism at the D2 receptor using PNU‐95666E resulted in bradycardia, hindquarter vasodilation and decreases in arterial pressure. Partial agonism at the D3 receptor with BP897 had no effect on heart rate, but did produce depressor responses driven by decreases in HQVR. Conversely, agonism of the D4 receptor using PD168077 resulted in modest hindquarter vasoconstriction that was not dose dependent. Hence, by comparison, agonism of the D4 receptor has little effect in the cardiovascular system of the rat relative to the other dopamine receptor subtype agonists tested.
Inhibition of inducible nitric oxide synthase reduces lipopolysaccharide‐induced renal injury in the rat
Authors: Kadkhodaee M , Qasemi A
Published: January 2005

1. Gram‐negative bacterial lipopolysaccharide (LPS) release and subsequent septic shock is a major cause of death in intensive care units. Lipopolysaccharide has been reported to increase the production of nitric oxide (NO) and the formation of oxygen‐derived free radicals (OFR) in different organs. The aim of the present study was to evaluate the role of an inducible form of NO synthase (iNOS) and OFR production in LPS‐induced renal impairment.

2. Measurement of vitamin E as the most important fat‐soluble anti‐oxidant was used as a marker of tissue oxidative stress. Lipopolysaccharide (10 mg/kg), l‐iminoethyl lysine (L‐Nil; 3 mg/kg, i.p.; a specific inhibitor of iNOS activity) and dimethyl thiourea (DMTU; 500 mg/kg i.p.; a well‐known OFR scavenger) were used. Four groups of eight rats were studied. One group received LPS, whereas a second group received LPS + L‐Nil. A third group received LPS + DMTU and the fourth group, receiving saline, acted as a control group. To evaluate renal function, plasma creatinine and blood urea nitrogen (BUN) were measured. High‐pressure liquid chromatography and ultraviolet detection were used to measure plasma and tissue vitamin E levels. Light microscopy was used to examine histopathological changes in the four groups.

3. Lipopolysaccharide markedly decreased the vitamin E content of renal plasma and tissue (P < 0.05). Administration of L‐Nil attenuated renal dysfunction and preserved vitamin E levels. However, DMTU failed to prevent renal injury, as indicated by plasma BUN levels and renal histology, despite the fact that it maintained renal vitamin E levels and increased plasma vitamin E levels. Thus, the overproduction of NO by iNOS may have a role in this model of LPS‐induced renal impairment.
Development of low‐affinity, membrane‐targeted Ca2+ sensors suitable for measuring presynaptic Ca2+
Authors: Monif M , Smart ML , Reid CA , Williams DA
Published: January 2005

1. Our aim is to measure near‐membrane Ca2+ flux within the presynaptic terminals of central neurons by modifying new genetically encoded Ca2+ sensors to develop tools capable of measuring localized Ca2+ signals.

2. We used standard recombinant DNA technologies to generate the DNA coding for a fusion construct of a modified fluorescent ‘pericam’ Ca2+ biosensor with a presynaptic P2X7 receptor (P2X7R). The Ca2+ sensitivity of the biosensor was modified by rational site‐directed mutagenesis of the calmodulin portion of the pericam.

3. Biosensor–receptor fusions were transfected into expression systems for evaluation. Expression studies in HEK‐293 cells showed that biosensor–receptor fusion construct‐delivered protein was localized exclusively to the plasma membrane, confirming that fusion did not affect the ability of the receptor to undergo normal protein synthesis and trafficking.

4. The Ca2+‐dependent fluorescence of the pericam portion of the fusion protein was also retained. Site‐direct mutagenesis within the calmodulin moiety of the pericam significantly reduced the Ca2+ affinity of the complex. The dynamic range of the sensor following this modification is better matched to the higher Ca2+ levels expected within presynaptic Ca2+ microdomains.
Functional imaging: New views on lens structure and function
Authors: Donaldson Paul J , Grey Angus C , Merriman‐Smith B Rachelle , Sisley Aran MG , Soeller Christian , Cannell Mark B , Jacobs Marc D
Published: January 2005

1. We have developed an experimental imaging approach that allows the distribution of lens membrane proteins to be mapped with subcellular resolution over large distances as a function of fibre cell differentiation.

2. Using this approach in the rat lens, we have localized precisely histological sites of connexin 46 cleavage, quantitatively mapped changes in gap junction distribution and fibre cell morphology and correlated these changes to differences in intercellular dye transfer.

3. Profiling of glucose transporter isoform expression showed that lens epithelial cells express GLUT1, whereas deeper cortical fibre cells express the higher‐affinity GLUT3 isoform. Near the lens periphery, GLUT3 was located in the cytoplasm of fibre cells, but it underwent a differentiation‐dependent membrane insertion.

4. Similarly, the putative adhesion protein membrane protein 20 is inserted into fibre cell membranes at the stage when the cells lose their nuclei. This redistribution is strikingly rapid in terms of fibre cell differentiation and correlates with a barrier to extracellular diffusion.

5. Our imaging‐orientated approach has facilitated new insights into the relationships between fibre cell differentiation and lens function. Taken together, our results indicate that a number of strategies are used by the lens during the course of normal differentiation to change the subcellular distribution, gross spatial location and functional properties of key membrane transport proteins.
Quantitative phase microscopy: A new tool for investigating the structure and function of unstained live cells
Authors: Curl Claire L , Bellair Catherine J , Harris Peter J , Allman Brendan E , Roberts Ann , Nugent Keith A , Delbridge Lea MD
Published: January 2005

1. The optical transparency of unstained live cell specimens limits the extent to which information can be recovered from bright‐field microscopic images because these specimens generally lack visible amplitude‐modulating components. However, visualization of the phase modulation that occurs when light traverses these specimens can provide additional information.

2. Optical phase microscopy and derivatives of this technique, such as differential interference contrast (DIC) and Hoffman modulation contrast (HMC), have been used widely in the study of cellular materials. With these techniques, enhanced contrast is achieved, which is useful in viewing specimens, but does not allow quantitative information to be extracted from the phase content available in the images.

3. An innovative computational approach to phase microscopy, which provides mathematically derived information about specimen phase‐modulating characteristics, has been described recently. Known as quantitative phase microscopy (QPM), this method derives quantitative phase measurements from images captured using a bright‐field microscope without phase‐ or interference‐contrast optics.

4. The phase map generated from the bright‐field images by the QPM method can be used to emulate other contrast image modes (including DIC and HMC) for qualitative viewing. Quantitative phase microscopy achieves improved discrimination of cellular detail, which permits more rigorous image analysis procedures to be undertaken compared with conventional optical methods.

5. The phase map contains information about cell thickness and refractive index and can allow quantification of cellular morphology under experimental conditions. As an example, the proliferative properties of smooth muscle cells have been evaluated using QPM to track growth and confluency of cell cultures. Quantitative phase microscopy has also been used to investigate erythrocyte cell volume and morphology in different osmotic environments.

6. Quantitative phase microscopy is a valuable, new, non‐destructive, non‐interventional experimental tool for structural and functional cellular investigations.
Microscopic imaging of extended tissue volumes
Authors: LeGrice Ian , Sands Greg , Hooks Darren , Gerneke Dane , Smaill Bruce
Published: January 2005

1. Detailed information about three‐dimensional structure is key to understanding biological function.

2. Confocal laser microscopy has made it possible to reconstruct three‐dimensional organization with exquisite resolution at cellular and subcellular levels.

3. There have been few attempts to acquire large image volumes using the confocal laser scanning microscope.

4. Previously, we have used manual techniques to construct extended volumes (several mm in extent, at 1.5 µm voxel size) of myocardial tissue.

5. We are now developing equipment and efficient automated methods for acquiring extended morphometric databases using confocal laser scanning microscopy.
T102C genetic polymorphism of the 5‐HT2A receptor in Chinese hypertensive patients and healthy controls
Authors: Yu Bang‐ning , Wang An , Zhou Gan , Zhang Wei , Hu Dong‐li , Li Qing , He Yi‐jing , Zhou Hong‐Hao
Published: January 2005

1. The 5‐HT2A receptor belongs to the G‐protein superfamily. It plays an important role in vascular regulation.

2. Previous reports in the UK have indicated that there is an association of the T102C genetic polymorphism of the 5‐HT2A receptor with hypertension, but no studies have been made on the T102C genetic polymorphism in Chinese hypertensive patients. In the present study, we investigated the T102C genetic polymorphism of 5‐HT2A receptors in Chinese hypertensive patients to determine whether there is an association of this polymorphism with hypertension in Chinese.

3. The present study was conducted on 198 hypertensive patients and 164 healthy controls. Polymerase chain reaction–restriction fragment length polymorphism was used to identify the T102C genetic polymorphism of the 5‐HT2A receptor.

4. In the present study, the C allele frequency of the 5‐HT2A receptor genetic polymorphism was 0.343 in hypertensive patients, which was not significantly different to that in healthy controls (0.393; χ2 = 1.922; P = 0.166; odds ratio = 0.807, 95% confidence interval 0.596–1.093). In addition, no gender differences were observed.

5. In conclusion, to our knowledge, this is the first report on the T102C genetic polymorphism of the 5‐HT2A receptor in Chinese hypertensive patients. We find that no correlation exists between the T102C genetic polymorphism and hypertension, which affords useful information on the pathogenesis of hypertension in the Chinese population.
Cardiac DT‐diaphorase contributes to the detoxification system against doxorubicin‐induced positive inotropic effects in guinea‐pig isolated atria
Authors: Badary Osama A , Awad Azza S , Abdel‐Maksoud Sahar , Hamada Farid MA
Published: January 2005

1. Doxorubicin (DOX), a standard chemotherapeutic anthracycline agent, causes a positive inotropic effect in guinea‐pig isolated atria in a concentration‐dependent manner with an ED50 of 3.6 µmol/L. This increase in contractility is strictly related to the generation of reactive oxygen species (ROS) as a consequence of quinone metabolism. The ED50 of DOX is significantly increased (P < 0.05) in the presence of 150 U superoxide dismutase (SOD). In the heart, DOX may be subjected to one‐ or two‐electron reductions catalysed by flavoenzymes in the presence of suitable electron donors. Two‐electron reduction is catalysed by NAD(P)H quinone acceptor oxidoreductase (DT‐diaphorase; DTD). Whether DOX will be activated or detoxified by two‐electron reduction is important for the understanding of the mechanism of both the toxic and antitumour actions of DOX.

2. In order to assess the role of DTD in cardiac responses to DOX, we examined the effect of both a specific inhibitor (dicoumarol) and an inducer (3‐methylcholanthrene; MCA) of the enzyme on the inotropic action of DOX.

3. In guinea‐pig isolated left atria, 4 µmol/L dicoumarol significantly enhanced the positive inotropic effect of DOX, especially at lower concentrations of DOX. In atria isolated from guinea‐pigs treated with MCA (44 mg/kg, i.p. for 4 days), DTD activity was enhanced (approximately twice that of the control; P < 0.01), whereas the activity of glutathione S‐transferase (GST) was not significantly altered. In these preparations, DOX caused a significantly lower increase in force of contraction than in atria isolated from untreated animals.

4. These results demonstrate that cardiac DTD does not contribute to ROS generation, but represents a detoxification system.
Role of non‐selective adenosine receptor blockade and phosphodiesterase inhibition in cisplatin‐induced nephrogonadal toxicity in rats
Authors: Saad Sherif Y , Najjar Tawfeeg AO , Alashari Mouied
Published: January 2005

1. It is well documented that cisplatin (CDDP) treatment increases the expression of adenosine A1 receptors in both kidney and testes. However, the effect of adenosine at these receptors is controversial. Adenosine A1 receptors have been documented to be involved in either cytoprotection or aggravation of nephrotoxicity. The aim of the present study was to examine the effect of the non‐selective adenosine receptor inhibitor theophylline and the phosphodiesterase inhibitor pentoxifylline on CDDP‐induced renal and testicular toxicity.

2. Male Wister rats were divided into six groups. Two control groups received plain drinking water and a third control group received theophylline 0.8 mg/mL in the drinking water for 2 weeks. One group of animals drinking plain water was injected intraperitoneally (i.p.) with pentoxifylline 50 mg/kg per day for 2 weeks. The remaining groups were treated in the same manner and received single dose of CDDP 7 mg/kg, i.p., 1 week after starting theophylline and pentoxifylline treatment and all animals were killed 1 week after CDDP treatment.

3. Rats treated with CDDP developed nephrotoxicity, as demonstrated by increased kidney and testes weight as a percentage of total bodyweight, blood urea nitrogen and serum creatinine levels and decreased serum calcium and albumin levels. In addition, CDDP treatment resulted in an increase in the production of malondialdehyde (MDA) and decreases in total nitrate/nitrite levels, as well as depletion of reduced glutathione (GSH) content and glutathione peroxidase (GPX) activity in both the kidney and testes. Administration of theophylline in the drinking water to CDDP‐treated rats resulted in exacerbation of the indices of nephrotoxicity, depletion of GSH content and GPX activity levels, with increased MDA production and platinum accumulation in both the kidney and testes. However, pentoxifylline administration reduced CDDP‐induced biochemical changes and reduced platinum accumulation in both organs. Histopathological examination of the kidney revealed that CDDP treatment produced multifocal tubular atrophy, atypical reparative changes of the tubular epithelium and marked tubular necrosis. Animals treated with the theophylline/CDDP combination showed extensive widespread damage with intratubular calcification. However, pentoxifylline treatment ameliorated the overt changes induced by CDDP treatment.

4. Theophylline exacerbates the deleterious effects of CDDP on rat kidney and testes. However, pentoxifylline alleviates CDDP‐induced renal and testicular toxicity.
Granulocyte colony stimulating factor/macrophage colony stimulating factor improves postinfarct ventricular function by suppression of border zone remodelling in rats‡
Authors: Miki Takayuki , Miura Tetsuji , Nishino Yasuhiro , Yano Toshiyuki , Sakamoto Jun , Nakamura Yuichi , Ichikawa Yoshihiko , Ikeda Yoshihiro , Kobayashi Hironori , Ura Nobuyuki , Shimamoto Kazuaki
Published: January 2005

1. The aim of the present study was to examine the effects of mobilization of bone marrow cells by granulocyte colony stimulating factor (G‐CSF) and macrophage colony stimulating factor (M‐CSF) on ventricular function after myocardial infarction (MI).

2. After ligation of the left coronary artery, rats were divided into a vehicle control group (MI group) and a CSF‐treated group (MI‐CSF group). Rats in the MI‐CSF group received a combination of G‐CSF (50 µg/kg per day) and M‐CSF (106 IU/kg per day) for 5 days after MI. Two weeks after MI, hearts were isolated and perfused with a Krebs' buffer and their functional responses to step‐wise elevation of left ventricular end‐diastolic pressure (LVEDP) were assessed. In histological analysis, proliferating cells and bone marrow‐derived cells were identified by antibodies against Ki‐67 and c‐kit and organization of collagen was examined by picrosirius red staining. The mRNA levels of transforming growth factor (TGF)‐β1, collagen type I and collagen type III were measured by quantitative reverse transcription–polymerase chain reaction.

3. Numbers of Ki‐67‐ and c‐kit‐positive cells in the infarct border zone after MI were increased by CSF treatment, but few of those cells were stained by anti‐α‐sarcomeric actin. The levels in mRNA of TGF‐β1 and collagen type I in the infarct border zone were higher in the CSF‐treated group compared with the MI group. Although CSF treatment did not reduce ventricular hypertrophy or infarct size at 2 weeks after MI, it did significantly improved the response of left ventricular developed pressure to step‐wise elevation of LVEDP. This effect was mimicked by treatment with M‐CSF alone. The functional improvement by CSF treatment was correlated with suppression of enlargement of the infarct–non‐infarct border associated with infarct expansion. Collagen fibres in the border zone were thicker and orientated more orderly in the CSF‐treated group than in the untreated group.

4. The results suggest that G‐CSF/M‐CSF treatment improves contractile function of the ventricle after infarction, presumably by acceleration of infarct repair and suppression of remodelling in the border zone.
Cardiac structure and electrical activation: Models and measurement
Authors: Smaill Bruce H , LeGrice Ian J , Hooks Darren A , Pullan Andrew J , Caldwell Bryan J , Hunter Peter J
Published: January 2005

1. Our group has developed finite element models of ventricular anatomy that incorporate detailed structural information. These have been used to study normal electrical activation and re‐entrant arrhythmia.

2. A model based on the actual three‐dimensional microstructure of a transmural left ventricular (LV) segment predicts that cleavage planes between muscle layers may give rise to non‐uniform, anisotropic electrical propagation and also provide a substrate for bulk resetting of the myocardium during defibrillation.

3. The model predictions are consistent with the results of preliminary experiments in which a novel fibre‐optic probe is used to record transmembrane potentials at multiple intramural sites in the intact heart. Extracellular potentials are recorded at adjacent LV sites in these studies.

4. We conclude that structural discontinuities in ventricular myocardium may play a role in the initiation of re‐entrant arrhythmia and discuss future studies that address this hypothesis.
Inherited cardiac arrhythmia syndromes: What have they taught us about arrhythmias and anti‐arrhythmic therapy?
Authors: Subbiah Rajesh N , Campbell Terence J , Vandenberg Jamie I
Published: January 2005

1. In recent years, the identification of the gene defects in a vast array of monogenic disorders has revolutionized our understanding of the basic mechanisms underlying numerous disease processes.

2. Mutations in cardiac ion channels have been identified as the basis of a wide range of inherited arrhythmia syndromes, including the congenital long QT syndromes, Brugada syndrome, Lenegre syndrome, Andersen's disease and familial atrial fibrillation.

3. Identification of mutations in the human‐ether‐a‐go‐go‐related gene (HERG) K+ channel as the molecular basis of congenital long QT syndrome type 2 also led to the discovery that HERG is the molecular target for the vast majority of drugs (both cardiac and non‐cardiac) that cause drug‐induced arrhythmias. This has had profound implications not only for the development of anti‐arrhythmic agents, but also for drug development in general.

4. The sequencing of the human genome in a sense represents the pinnacle of the reductionist era of molecular medicine. The great challenge now is to re‐integrate the information gathered during the ‘reductionist era’ to provide a better understanding of the intact organism. Computer modelling is likely to be a key component of that re‐integration process.
Bladder cancer discovered by ovarian metastasis: Cytokeratin expression is useful when making differential diagnosis
Authors: ISHII YASUHIKO , ITOH NAOKI , TAKAHASHI ATSUSHI , MASUMORI NAOYA , IKEDA TATSURU , TSUKAMOTO TAIJI
Published: January 2005

Abstract A 49‐year‐old woman underwent hysterectomy and bilateral adnexectomy after the diagnosis of a right ovarian tumor with paraaortic and pelvic lymph node metastases. The pathological diagnosis was undifferentiated carcinoma of the ovary. After the operation, a bladder tumor was discovered during the evaluation for microscopic hematuria. The bladder tumor was pathologically diagnosed as transitional cell carcinoma, pT1b, G3. Although the pathological findings of the bladder cancer and ovarian cancer were very similar, we could diagnose primary bladder cancer with ovary and lymph node metastases according to the immunohistochemical staining pattern of cytokeratins 7 and 20. Herein, the clinical usefulness of immunohistochemical staining using cytokeratins for making a differential diagnosis of the origin of a tumor in the pelvic cavity is demonstrated.
Genistein induces cell growth inhibition in prostate cancer through the suppression of telomerase activity
Authors: OUCHI HIDEKI , ISHIGURO HITOSHI , IKEDA NAOYA , HORI MAYUMI , KUBOTA YOSHINOBU , UEMURA HIROJI
Published: January 2005

Abstract Aim:  To clarify the mechanism of the anticancer effect of genistein, we examined the effect of genistein on telomerase activity in prostate cancer cells. We hypothesized that genistein may exert its anticancer effect by modifying telomerase activity in prostate cancer cells.

Methods:  Prostate cancer (LNCaP) cells were cultured with genistein and the number of viable cells was counted. Growth medium was also collected to measure prostate‐specific antigen (PSA) concentration. Polymerase chain reaction (PCR)‐based telomeric repeat amplification protocol (TRAP) assay and reverse transcriptase (RT)‐PCR analysis were performed to investigate telomerase activity and the expression of human telomerase reverse transcriptase (hTERT), c‐myc and p21 mRNA. To examine the possibility that hTERT transcriptional activity is modulated by genistein, transient cell transfection studies were performed by using luciferase reporter assay. Telomere repeat amplification protocol (TRAP) assay and PCR analysis of hTERT were performed in androgen independent cells, DU‐145.

Results:  Cell growth of LNCaP was inhibited by genistein and PSA secretion was similarly reduced. In TRAP assay, the telomerase activity of LNCaP cells was reduced by genistein. Reverse transcriptase‐PCR analysis revealed that the expression of hTERT and c‐myc mRNA was down‐regulated by genistein, whereas p21 mRNA increased in response to genistein. Luciferase reporter assay revealed that genistein reduced the transcriptional activity of hTERT. In DU‐145 cells, telomerase activity and the expression of hTERT mRNA were also reduced by genistein.

Conclusion:  The current study elucidated the molecular mechanism of cell growth inhibition by genistein. The antiproliferative effects of genistein seem to be exerted on the hTERT transcriptional activity via different molecular pathways.
Bilateral ureteral fibroepithelial polyps and review of the literature
Authors: TEKDOGAN UMIT YENER , CANAKLI FATIH , ASLAN YILMAZ , HAN OZGE , GUNGOR SEFA , ATAN ALI
Published: January 2005

Abstract Despite being the most common forms of benign ureteral neoplasms, ureteral fibroepithelial polyps are rare and their etiology is still unknown. To our knowledge, we report the fourth case of bilateral fibroepithelial polyps in the English literature with a discussion of clinical features and etiology.
Torsion of a communicating hydrocele in a child
Authors: SHIRAISHI KOJI , MOHRI JUN , EGUCHI SATOSHI , KAMIRYO YORIAKI , UEKI KOJI
Published: January 2005

Abstract Among the diseases presenting as acute scrotum, torsion of a communicating hydrocele is extremely rare. A 5‐year‐old boy was referred with discomfort and swelling of the left scrotum. Operative findings revealed a torsion of a communicating hydrocele. We report a case of torsion with preoperative findings of ultrasonography and magnetic resonance imaging. We also emphasize the awareness of this disease as a differential diagnosis of the acute scrotum.
Solitary psoas muscle metastasis after radical nephrectomy for renal cell carcinoma
Authors: TAIRA HIROSHI , ISHII TATSU , INOUE YOSHIO , HIRATSUKA YOSHIHARU
Published: January 2005

Abstract Skeletal muscle is a very rare location for the metastasis of renal cell carcinoma (RCC) and only one case of solitary metastasis to the psoas muscle has been reported. We present a 63‐year‐old male patient with late recurrence (14 years) after left side radical nephrectomy for RCC. He first visited Chikushi Hospital, Fukuoka University, Japan in January 2000 for a postoperative follow‐up because he had shifted residence to the area. Follow‐up was by abdominal computed tomography (CT) and chest X‐ray. In December 2001, a CT scan showed a 1.5 cm enhanced mass in the right psoas muscle without any other metastasis. The mass was resected that month and histological study showed RCC metastasis.
Small cell carcinoma of the prostate with hypercalcemia
Authors: HANAZAWA KISABURO , HIGASHI NAOTAKA , KAWACHI YOSHIO , SUZUKI FUJIHIKO , ISHI KAZUHISA , FUJIME MAKOTO
Published: January 2005

Abstract We present a case of small cell prostate carcinoma with hypercalcemia in a 75‐year‐old man. He was diagnosed as having stage T3bN1M0 adenocarcinoma of the prostate. His serum prostate‐specific antigen level was reduced to below the normal range after a combination treatment of a luteinizing hormone‐releasing hormone agonist and flutamide for prostate carcinoma. He subsequently experienced increasing fatigue, poor appetite, short time loss of consciousness and pain in his lower abdomen. His serum calcium level and carcinoembryonic antigen were increased. He died 5 months from the start of treatment. The autopsy revealed small cell carcinoma of the prostate and multiple metastasis of the lung, liver, pancreas, lymph nodes and spine.