bluuebunny/crossref_metadata_2025 huggingface dataset:
Redox side reactions of haemoglobin and cell signalling mechanisms1
Authors: Yeh L.‐H. , Alayash A. I.
Published: December 2004

Abstract.  Yeh L‐H, Alayash AI (Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, MD, USA). Redox side reactions of haemoglobin and cell signalling mechanisms (Minisymposium). J Intern Med 2003; 253: 518–526.

Cell‐free chemically modified or recombinant haemoglobins developed as oxygen therapeutics are designed to correct oxygen deficit caused by ischaemia in a variety of clinical settings. Oxidative processes, which are in some cases enhanced when modifications are introduced that lower oxygen affinity, can limit the safety of these proteins. Direct cytotoxic effects associated with haemoglobins have been ascribed to the redox reactions between haemoglobin and biological peroxides [i.e. hydrogen peroxide (H2O2), lipid peroxides (LOOH) and peroxynitrite (ONOO–)]. Biochemical changes at the cellular, tissue and organ levels have been documented to occur in response to haemoglobin oxidative reactions. These peroxides have been implicated as regulators of redox sensitive cell signalling pathways. The effects of reactions between haemoglobin and biologically relevant peroxides may be more subtle than oxidative damage and may thus involve perturbation of redox sensitive signalling pathways. In this review, a brief outline of the role of cell‐free haemoglobin in oxidative and cell‐signalling pathways and the implications of these reactions on the safety and efficacy evaluation of haemoglobin‐based oxygen carries are presented.
Effect of vitamins and aspirin on markers of platelet activation, oxidative stress and homocysteine in people at high risk of dementia
Authors:   VITAL Trial Collaborative Group
Published: December 2004

Abstract. VITAL Trial Collaborative Group. Effect of vitamins and aspirin on markers of platelet activation, oxidative stress and homocysteine in people at high risk of dementia. J Intern Med 2003; 254:–.

Objectives. To examine the association of cognitive impairment with platelet activation and reactive oxygen species and total homocysteine levels; and to assess the biochemical efficacy of treatment with aspirin and vitamin supplements in people at high risk of dementia.

Subjects. People with dementia or mild cognitive impairment.

Design and intervention. In a 2 × 2 × 2 factorial design trial, 149 people at high‐risk of dementia were randomized to receive either low‐dose aspirin (81 mg) or placebo; and folic acid (2 mg) plus vitamin B12 (1 mg) or placebo; and vitamins E (500 mg) plus C (200 mg) or placebo. Participants were seen twice before and once after 12 weeks of treatment.

Main outcome measures. At each visit, participants had their cognitive function assessed and had blood collected for homocysteine, folate and vitamin B12 determination and urine collected for markers of platelet activation (11‐dehydro‐thromboxane B2) and reactive oxygen species (8‐epi‐PGF2α).

Results. Prior to treatment, cognitive function was inversely related with homocysteine and with urinary thromboxane and isoprostane, and these associations were independent of age. Aspirin was associated with a median reduction in 11‐dehydrothromboxane B2 of 73% (P < 0.001). B‐vitamins lowered plasma homocysteine concentration by 30% (P < 0.0001) and antioxidant vitamins lowered isoprostane excretion by 26% (P < 0.1). No effect of treatment on cognitive function was detected.

Conclusions. Aspirin and B‐vitamins were effective in reducing biochemical factors associated with cognitive impairment in people at risk of dementia. Large‐scale trials are now required to assess the relevance of aspirin and B‐vitamins for the maintenance of cognitive function in people at risk of dementia.
F‐18‐fluorodeoxyglucose positron emission tomography leading to a diagnosis of septic thrombophlebitis of the portal vein: description of a case history and review of the literature
Authors: Bleeker‐Rovers C. P. , Jager G. , Tack C. J. , Van Der Meer J. W. M. , Oyen W. J. G.
Published: December 2004

Pylephlebitis or septic thrombophlebitis of the portal vein is a serious infectious disorder. Early diagnosis is difficult, due to nonspecific symptoms and signs, limitations of diagnostic modalities and the lack of familiarity of physicians with this entity. We report the history of a 73‐year‐old man with fever of unknown origin (FUO) in whom laboratory tests, blood and urine cultures, chest X‐ray, abdominal ultrasound, and Indium‐111‐leucocyte scintigraphy did not reveal the cause of the fever. F‐18‐fluorodeoxyglucose (FDG) positron emission tomography (PET) subsequently pointed to the diagnosis of pylephlebitis, which was confirmed by computed tomography (CT) and percutaneous puncture. We conclude that FDG PET allows detecting inflammatory foci in patients with FUO and offers to make the diagnosis of pylephlebitis at an early stage.
Translation of clinical trials into clinical practice
Authors: Julian D. G.
Published: December 2004

The results of well‐conducted clinical trials should be translated into practice but there is good evidence that this is not happening. There are a number of reasons for this – ignorance of doctors and patients, uncertainty as to the applicability of trials to individual patients, indolence and inefficiency on the part of practitioners, and financial considerations. More attention needs to be paid to correct all these factors.
The behaviour of doramectin in the gastrointestinal tract, its secretion in bile and pharmacokinetic disposition in the peripheral circulation after oral and intravenous administration to sheep
Authors: HENNESSY D. R. , PAGE S. W. , GOTTSCHALL D.
Published: December 2004

Sheep were ‘compartmentalized’ by surgically implanting cannulae in the rumen, abomasum and terminal ileum with a re‐entrant cannula inserted between the cystic duct and the duodenum to monitor bile secretion. Doramectin, containing a trace of [3H]‐doramectin, was administered both intravenously (i.v.) and intraruminally (i.r.) at a dosage of 150 μg/kg. The pharmacokinetic behaviour of [3H]‐labelled products was determined in these pools, and also in peripheral plasma, urine and faeces. Parent doramectin was also determined in plasma, abomasal digesta fluid and bile. Following i.r. administration, [3H] compounds were almost entirely associated with particulate digesta. A 14.5‐h half‐life in the rumen prolonged the presence of [3H] in the abomasum. Doramectin appeared to be degraded in abomasal digesta because only 24% of abomasal [3H] was attributed to the parent drug. Absorption of doramectin resulted in a systemic availability of 35%, of which 1.6 and 23.6% of the dose was contained in urine and biliary secretions, respectively. Following i.v. administration, almost negligible quantities of [3H] were secreted into the rumen or abomasum and only 2.7% of the dose was excreted in urine, whereas 132% was secreted in bile. This indicated that approximately one‐third of biliary metabolites were enterohepatically recycled with biliary metabolites, elevating the proportion of [3H] in fluid digesta in the small intestine. Passage of the IR‐administered drug through the gastrointestinal tract (GIT) resulted in virtually complete faecal excretion of [3H] within 5 days, whereas the continued secretion of i.v.‐administered [3H] in bile prolonged the presence of [3H] in the GIT, with faecal clearance not being complete for at least 10 days. This multi‐compartmental study has provided more information on the behaviour of doramectin than can be obtained from examining drug disposition in the peripheral circulation alone. With this knowledge, it is anticipated that opportunities for improving drug performance will be identified.
In vitroandex vivoeffects of the phosphodiesterase 4 inhibitor, rolipram, on thromboxane production in equine blood
Authors: Rickards K. J. , Page C. P. , Lees P. , Gettinby G. , Cunningham F. M.
Published: December 2004

Phosphodiesterase 4 (PDE4) inhibitors have been shown to inhibit equine neutrophil functionin vitroand may be of benefit in recurrent airway obstruction (RAO), an allergy‐based respiratory disease characterized by inflammatory cell recruitment and activation within the lungs following exposure of susceptible horses to allergens in mouldy hay. The aim of this study was to evaluate the inhibitory effects of the PDE4 inhibitor, rolipram, in anin vitroassay of thromboxane (Tx) production. The assay was then used to monitor the activity of this compoundin vivoin normal and RAO‐affected horses. Rolipram and the structurally distinct PDE4 inhibitor, denbufylline, attenuated both lipopolysaccharide (LPS)‐induced and unstimulated Tx production in blood from normal horses. Thromboxane production appeared to involve a calcium‐dependent interaction between leucocytes and platelets (LPS‐induced Tx production = 2.3 ± 0.4, 4.5 ± 1.1 and 20.8 ± 3.6 ng/mL for platelets, leucocytes and blood, respectively) and rolipram‐inhibited Tx production via an effect on leucocytes. Inhibition ofex vivoLPS induced Tx production was detected after intravenous administration of rolipram (5 μg/kg) to normal ponies. This dose did not significantly affect either lung function or neutrophil accumulation when administered to three horses with clinical signs of RAO. This study suggests that inhibition of Tx production in equine blood can be used to measure PDE4 activity. However, PDE4 inhibitors with improved therapeutic profiles are required for evaluation in RAO.
Antimicrobial resistance in livestock
Authors: Catry B. , Laevens H. , Devriese L.A. , Opsomer G. , de Kruif A.
Published: December 2004

Antimicrobial resistance may become a major problem in veterinary medicine as a consequence of the intensive use and misuse of antimicrobial drugs. Related problems are now arising in human medicine, such as the appearance of multi‐resistant food‐borne pathogens. Product characteristics, dose, treatment interval and duration of treatment influence the selection pressure for antimicrobial drug resistance. There are theoretical, experimental and clinical indications that the emergence of de novo resistance in a pathogenic population can be prevented by minimizing the time that suboptimal drug levels are present in the infected tissue compartment.

Until recently, attention has been focused on target pathogens. However, it should be kept in mind that when antimicrobial drugs are used in an individual, resistance selection mainly affects the normal body flora. In the long term, this is at least equally important as resistance selection in the target pathogens, as the horizontal transfer of resistance genes converts almost all pathogenic bacteria into potential recipients for antimicrobial resistance. Other factors contributing to the epidemiology of antimicrobial resistance are the localization and size of the microbial population, and the age, immunity and contact intensity of the host. In livestock, dynamic herd‐related resistance patterns have been observed in different animal species.
Azithromycin metabolite identification in plasma, bile, and tissues of the ball python (Python regius)
Authors: Hunter R. P. , Koch D. E. , Coke R. L. , Goatley M. A. , Isaza R.
Published: December 2004

Azithromycin is the first of a class of antibiotics classified as azalides. Six ball pythons (Python regius) were given a single dose of azithromycin at 10 mg/kg p.o. and i.v. in a crossover design. Serial blood samples were collected for unchanged azithromycin and to determine, if possible, the structure and number of circulating azithromycin metabolites. After a 4‐month wash‐out period, the snakes were given azithromycin p.o. as a single dose of 10 mg/kg for the study of azithromycin metabolism and metabolite tissue distribution. Bile, liver, lung, kidney, and skin samples were analyzed for the metabolites identified from the first experiment. Unchanged azithromycin accounted for 80, 68, and 60% of the total material at 12, 24, and 48 h postadministration in plasma, independent of route of administration. At both 24 and 72 h postadministration, azithromycin accounted for 70% of total azithromycin‐ associated material in bile. In liver and kidney, unchanged azithromycin accounted for 40% of the total azithromycin‐associated material; this doubled in lung and skin. Fifteen metabolites were positively or tentatively identified in plasma, bile, or tissues of all snakes. Four of these possible metabolites: 3′‐desamine‐3‐ene‐azithromycin, descladinose dehydroxy‐2‐ene‐azithromycin, 3′‐desamine‐3‐ene descladinose‐azithromycin, and 3′‐N‐nitroso,9a‐N‐desmethyl‐azithromycin are unique to this species. Descladinose‐azithromycin, 3′‐N‐desmethyl,9a‐N‐desmethyl‐azithromycin, and 3′‐N‐desmethyl, 3′‐O‐desmethyl‐azithromycin were the only metabolites identified in skin. Kidney tissue contained a greater number of metabolites than liver tissue, with 3′‐N‐didesmethyl‐azithromycin being identified only in the kidney. Compared with the dog and cat, a greater number of metabolites were identified in ball python plasma. The percentage of unchanged azithromycin in bile is not different between the three species.
The role of intrahepatic immune effector cells in inflammatory liver injury and viral control during chronic hepatitis B infection
Authors: Tang T. J. , Kwekkeboom J. , Laman J. D. , Niesters H. G. M. , Zondervan P. E. , de Man R. A. , Schalm S. W. , Janssen H. L. A.
Published: December 2004

Summary. Cytotoxic T lymphocytes (CTL) and Kupffer cells play an important role in the immune control of hepatitis B virus (HBV), but may also induce liver injury during infection. We investigated the intrahepatic immune response in liver biopsies of chronic HBV patients in relation to inflammatory liver injury and viral control. Forty‐seven liver biopsies from patients with chronic HBV with varying degrees of inflammation (ALT values) were selected. Acute hepatitis and normal liver specimens served as controls. Immune effector cells, cytotoxic effector molecules and cytokine producing cells were quantified after immunohistochemical staining in lobular and portal areas of the biopsies. The intralobular number of CD8+ T‐lymphocytes was significantly decreased in biopsies of patients with high ALT (r = −0.54; P < 0.001). Higher ALT‐values were correlated with increased numbers of granzyme+ cells in portal areas (r = 0.65; P < 0.001) and higher numbers of intralobular Fas‐L+ cells (r = 0.32; P = 0.05). Fas‐L was expressed on Kupffer and lymphoid cells. More intralobular CD8+ T‐lymphocytes were found in HBeAg− than in HBeAg+ patients (P = 0.002). But IFN‐γ and TNF‐α producing cells were observed sporadically in chronic HBV patients. Hence, in chronic HBV infection, low viral replication and HBeAg negativity is related to increased presence of intralobular CD8+ T‐lymphocytes. Persistence of the virus may be caused by the absence of cells producing anti‐viral cytokines in the liver. Inflammatory liver injury during chronic HBV infection is probably not the result of increased numbers of infiltrating CD8+ T‐lymphocytes, but of Fas‐L expression by Kupffer cells and increased cytolytic activity of cells in portal areas.
Aetiology and prognostic factors in acute liver failure in India
Authors: Khuroo M. S. , Kamili S.
Published: December 2004

Summary.  The early prognostic indicators for acute liver failure in endemic zones for hepatitis E virus have not been determined. All consecutive patients with acute liver failure from a geographically defined region endemic for hepatitis E virus were studied over the period April 1989–April 1996. Demographic, clinical and biochemical parameters were recorded at presentation and serum samples were analysed for known viral hepatitis (A–E) markers. Multiple parameters were compared in survivors and non‐survivors in a univariate analysis. All significant factors on univariate analysis were entered into a stepwise logistic regression analysis to identify independent variables of prognosis. The sensitivity and specificity of significant prognostic factors was then assessed.

A total of 180 [69 males and 111 females: age (mean ± SD) 31.1 ± 14.7 years] with acute liver failure were studied. Of these, 131 (72.8%) patients died. Hepatitis E virus was the aetiological cause in 79 (43.9%) patients, while hepatitis A virus, hepatitis B virus, hepatitis C virus and non‐A, non‐E agent/'s could be incriminated in four (2.1%), 25 (13.9%), 13 (7.2%) and 56 (31.1%) patients respectively. Of 83 women in childbearing age, 49 (59.0%) were pregnant, 33 (67.3%) of these were in the third trimester. Forty‐seven (95.8%) pregnant women had HEV infection. Nine variables differed significantly between survivors and non‐survivors on univariate analysis. Of these, four variables which predicted the adverse outcome on multivariate analysis were non‐hepatitis‐E aetiology, prothrombin time >30 s, grade of coma >2 and age >40 years in that order of significance. Pregnancy per se or duration of gestation did not adversely affect the prognosis. In endemic areas, hepatitis E virus is the commonest cause of acute liver failure. Acute liver failure occurs in a high proportion of pregnant women, mostly in third trimester. Early predictors of a poor outcome are non‐E aetiology, prothrombin time >30 s, grade of coma >2 and age >40 years.
A rapid immunochromatographic assay for hepatitis B virus screening
Authors: Lau D. T.‐Y. , Ma H. , Lemon S. M. , Doo E. , Ghany M. G. , Miskovsky E. , Woods G. L. , Park Y. , Hoofnagle J. H.
Published: December 2004

Summary. Simple, rapid and accurate assays for hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg) are helpful for clinical diagnosis and field epidemiological surveys. A commercially developed, rapid immunochromatographic test for simultaneous detection of HBsAg and HBeAg was evaluated using a total of 2463 selected samples (827 frozen sera, 1011 fresh sera, and 625 whole blood samples). Results of the rapid test were compared with standard enzyme immunoassay (EIA) methods for HBsAg and HBeAg detection. The accuracy of the rapid test was excellent and was similar for frozen sera, fresh sera and whole blood. The overall sensitivity and specificity for the detection of HBsAg were 95 and 100%, and the corresponding positive and negative predictive values were 100 and 99.7%, respectively. The sensitivity and specificity for the detection of HBeAg were slightly less than that for HBsAg, and were 80 and 98%, with positive and negative predictive values of 91 and 94%, respectively. Thus, compared with the EIA method, the rapid test was highly sensitive and accurate for the detection of HBsAg although somewhat less sensitive and specific for detection of HBeAg. Because of its speed, simplicity and flexibility, the rapid test is ideally suited for HBsAg and HBeAg screening in population‐based epidemiological studies and in low risk populations, particularly in regions of the world where hepatitis B is endemic.
Who should receive hepatitis A vaccine?
Authors: Arankalle V. A. , Chadha M. S.
Published: December 2004

Summary. Though a potent vaccine represents a powerful preventive tool, the policy of its use is governed by epidemiological and economical factors. Hepatitis A, an enterically trasmitted disease shows distinct association with socio‐economic status, populations with improvement experiencing lower exposure to the virus. With the availability of vaccine, it is pertinent to consider its use in the effective control of the disease. However, with the varied epidemiological patterns and economical constraints in different countries it does not seem to be possible to evolve universal policy for immunization. Though, universal immunization may be the most effective way of control, the same is not practical for many countries. It is proposed that irrespective of endemicity of hepatitis A, high‐risk groups such as travelers to endemic areas, patients suffering from chronic liver diseases, HBV and HCV carriers, tribal communities with high HBV carrier rates, food handlers, sewage workers, recipients of blood products, troops, and children from day‐care centers should be immunized with hepatitis A vaccine. In addition, for populations with intermediate prevalence, infants, children from affordable families may be immunized. As coupling the vaccine with EPI schedule would be beneficial, use of combined A & B or A, B & E vaccine may be an attractive alternative.
Retreatment with interferon and ribavirin vs interferon alone according to viraemia in interferon responder‐relapser hepatitis C patients: a prospective multicentre randomized controlled study
Authors: Portal I. , Bourlière M. , Halfon P. , De Lédinghen V. , Couzigou P. , Bernard P. H. , Blanc F. , Caroli‐Bosc F. , Arpurt J. P. , Vetter D. , Mathieu‐Chandelier C. , Chazouillères O. , Thiefin G. , Pol S. , Sogni P. , Abergel A. , Bailly F. , Picon M. , Debonne J. M. , Zamora C. , Alleman I. , Moreau X. , Doll F. , Eugène C. , Ducloux S. , Larrey D. , Ouzan D. , Grimaud J. C. , Gouvernet J. , Botti G. , Gérolami V. , Khiri H. , Gérolami A. , Gauthier A. P. , Botta‐Fridlund D.
Published: December 2004

Summary.  Low pretreatment viral load has consistently been shown to be an independent predictor of sustained response (SR) in patients with chronic hepatitis C infection. We assessed the efficacy of interferon (IFN) plus ribavirin vs IFN alone in low viraemic patients (<2 millions copies/mL) who had relapsed to a previous course of IFN and the efficacy of 24 vs 48 week combination therapy in high viraemic patients. Two hundred and ninety‐seven patients were randomly assigned to one of the four regimens after stratification on pretreatment viral load. All patients received IFN‐α2b (6 million units thrice weekly for 24 weeks and 3 million units thrice weekly for 24 weeks). Patients with low viraemia received either IFN‐α2b alone for 48 weeks (R1: 42 patients) or IFN‐α2b plus ribavirin (600 mg/day) for 24 weeks and IFN‐α2b alone for the next 24 weeks (R2: 48 patients). Patients with high viral load received either IFN‐α2b plus ribavirin for 24 weeks and then IFN‐α2b alone for the next 24 weeks (R3: 104 patients) or IFN‐α2b plus ribavirin for 48 weeks (R4: 103 patients). In low viraemic patients the rate of SR was 37.7% in group R1 and 59.6% in group R2 (P < 0.05). In high viraemic patients, the rate of SR was 44.7% in group R3 and 51.4% in group R4 (P: NS). Thirty‐one patients discontinued treatment (10.4%) without difference regarding treatment regimen. In the regimen using ribavirin we found no difference in terms of SR between patients receiving a dose of ribavirin below 10.6 mg/kg/day (55%) or over 10.6 mg/kg/day (58%). Histological improvement occurred in 70.2% of patients regardless of the regimen. Logistic regression showed that genotype 2 and 3, Knodell score <6 and alanine aminotransferase pretreatment level >3 × upper limit of normal were significantly and independently correlated with SR. In low viraemic patients who relapsed to a previous IFN treatment, combination therapy using high‐dose IFN and low‐dose ribavirin is better than high‐dose IFN alone. In high viraemic patients there was no benefit in increasing the duration of combination therapy from 24 to 48 weeks. In this study, it was found that low dose of ribavirin can be used safely and there is no effect of ribavirin dose on SR.
Reinforced interferon alpha‐2b and ribavirin is more effective than standard combination therapy in the retreatment of chronic hepatitis C previously nonresponsive to interferon: a randomized trial
Authors: Poynard T. , Marcellin P. , Bissery A. , Myers R. P. , Moussalli J. , Degos F. , Dhumeaux D. , Riachi G. , Bronowicki J. P. , Brissot P. , Buffet C. , Serfaty L. , Naveau S. , Sogni P. , Beaugrand M. , Gayno S. , Larrey D. , Samuel D. , Eugene C. , Pol S. , Bedossa P. , Daurat V. , Chaumet‐Riffaud P. ,   for the multicenter GER‐CYT‐RIBANON group
Published: December 2004

Summary. Interferon‐alpha (IFN) monotherapy results in sustained virological clearance in a minority of patients with chronic hepatitis C. The aim of this study was to assess the effect of a reinforced regimen combining ribavirin and high‐dose IFN for 48 weeks compared with a nonreinforced regimen combining a standard IFN regimen and ribavirin for 24 weeks in nonresponders with chronic hepatitis C. A total of 231 patients with chronic hepatitis C and previous nonresponse to IFN monotherapy were randomized. The reinforced group (n = 114) received IFN‐2b 6 million units (MU) thrice weekly (TIW) and ribavirin for 48 weeks, and the nonreinforced group (n = 117) received IFN‐2b 3 MU TIW and ribavirin for 24 weeks. The main outcome measure was a sustained virological response, defined as negative serum hepatitis C virus (HCV)‐RNA 24 weeks following the end of treatment. This endpoint was determined in 98 patients of the reinforced group and 105 patients of the nonreinforced group. At the end of follow‐up, a sustained virological response was observed in 29 of the 98 patients (29.6%) in the reinforced group vs 16 of the 105 patients (15.2%) in the nonreinforced group (P = 0.014). In multivariate analysis, factors associated with a sustained virological response were treated with a reinforced regimen [odds ratio (OR) 2.9; P = 0.06] and genotype 2 or 3 (OR 8.8; P < 0.0002). A total of 160 patients had paired biopsies before and after treatment. Histological activity improvement was observed in 32 of 80 patients (40%) and fibrosis worsening in 26 of 80 patients (33%) in the reinforced group vs 13 of 80 (16%) and 19 of 80 (24%) in the nonreinforced group (P =  0.30 and 0.20, respectively). Hence in nonresponders, a high‐dose 48‐week regimen of IFN and ribavirin combination was more effective than a regimen with interferon at lower dose and ribavirin for 24 weeks only.
Clinical relevance of total HCV Core antigen testing for hepatitis C monitoring and for predicting patients’ response to therapy
Authors: Maynard M. , Pradat P. , Berthillon P. , Picchio G. , Voirin N. , Martinot M. , Marcellin P. , Trepo C.
Published: December 2004

Summary. To study the correlation between total Hepatitis C virus (HCV) Core antigen (Ag) and HCV‐RNA, and to assess the proficiency of HCV Core Ag testing in monitoring and predicting virologic response during and after pegylated interferon (PEG‐IFN) and ribavirin combination therapy.

A total of 307 samples from treated and untreated patients were used to assess the correlation between the total HCV Core Ag test and quantitative HCV‐RNA assays (Superquant, and Quantiplex branched DNA 2.0 assay). Twenty‐four patients received combination therapy for 48 weeks. Blood samples were collected at day 0, and week 2, 4, 12, 24, 48 and 72 for virologic evaluation.

A linear relation exists between total HCV Core Ag and HCV‐RNA levels. At 3 months the positive predictive value (PPV) of response to therapy was 100% with either HCV Core Ag or HCV‐RNA. For HCV Core Ag the negative predictive value (NPV) was 100% whereas for HCV‐RNA the NPV was 80% (P > 0.05). At month 1, the PPV was 95% and 100% when determined by HCV Core Ag and HCV‐RNA, respectively. The NPV value was 100% for HCV Core Ag and 33% for HCV‐RNA (P = 0.005).

HCV Core Ag quantification could be useful in clinical practice to predict a sustained virological response early during therapy (4 weeks), reaching an optimal performance at month 3. The determination of total HCV Core Ag levels in serum, constitutes an accurate and reliable alternative to HCV‐RNA for monitoring and predicting treatment outcome in patients receiving PEG‐IFN/Ribavirin combination therapy.
Liver p53 expression in patients with HCV‐related chronic hepatitis
Authors: Loguercio C. , Cuomo A. , Tuccillo C. , Gazzerro P. , Cioffi M. , Molinari A. M. , Del Vecchio Blanco C.
Published: December 2004

Summary. Mutated p53 acts as a dominant oncogene and alterations in the p53 gene are described in a large number of patients with hepatocellular carcinoma (HCC). It has been demonstrated that hepatitis C virus (HCV)‐core protein regulates transcriptionally cellular genes, as well as cell growth and apoptosis. This study was undertaken to evaluate whether p53 may be expressed also in a precocious stage of HCV‐related liver damage.

We studied p53 expression by immunoluminometric assay on liver samples from 40 patients (M/F 18/ 22, median age 44 years, range 13–64 years) with biopsy‐proven HCV‐related chronic hepatitis. We considered the following factors: degree of liver damage, liver iron content and HCV‐RNA titre. We also evaluated as possible co‐factors alcohol and food intake in the last 3 years.

p53 was over‐expressed in seven of 40 (17.5%) patients. Liver histology documented the presence of unexpected cirrhosis in two patients among the p53 positive subjects. The p53 positive group had a daily ethanol intake significantly higher in respect to that of the p53 negative group (P < 0.05). Alimentary history documented that patients with a p53 over‐expression had a lower intake of total calories, monounsaturated fatty acids, vitamin C and riboflavin.

Data indicate that p53 over‐expression can occur even in initial stages of HCV‐related liver disease.
Dual effects of hepatitis C virus Core protein on the transcription of cyclin‐dependent kinase inhibitor p21 gene
Authors: Kwun H. J. , Jang K. L.
Published: December 2004

Summary. Transcription of p21 was activated in hepatitis C virus (HCV) Core‐expressing HepG2 cells where its upstream p53 was stabilized. However, this effect was not absolutely required for the activation of p21 by Core, as demonstrated in Hep3B cells. In addition, an opposite effect on the transcription of p21 was observed in NIH3T3 and primary hepatocytes, where p53 was not decreased by Core. To explain the p53‐independent regulation of p21 by Core, we identified a Core‐responsive element between positions −74 and −83 of the p21 promoter, exactly overlapped with a tumour growth factor β (TGF‐β)/butyrate responsive element. Furthermore, we demonstrated that Core could activate the p21 through the element by stimulating a butyrate pathway, whereas this was inhibited through a TGF‐β pathway. The opposing effects of Core protein on the transcription of p21 might be important in understanding the progression of hepatic disease in HCV‐positive patients.
Clinical evaluation of a new enzyme immunoassay for hepatitis B virus core‐related antigen; a marker distinct from viral DNA for monitoring lamivudine treatment
Authors: Rokuhara A. , Tanaka E. , Matsumoto A. , Kimura T. , Yamaura T. , Orii K. , Sun X. , Yagi S. , Maki N. , Kiyosawa K.
Published: December 2004

Summary. We aimed to assess the clinical performance of a newly developed chemiluminescence enzyme immunoassay (CLEIA) for the detection of hepatitis B virus (HBV) core‐related antigen (HBcrAg) in patients with chronic HBV infection. A total of 82 patients with chronic HBV infection and 167 HBV‐negative controls were studied. HBcrAg was measured by CLEIA with monoclonal antibodies to hepatitis B e antigen (HBeAg) and hepatitis B core antigen (HBcAg), and HBV DNA was measured by transcription‐mediated amplification assay (TMA) and in‐house real‐time detection polymerase chain reaction (RTD‐PCR). The HBcrAg assay detected viremia in 189 of 216 samples (88%) collected from 72 patients whilst the TMA assay detected viremia in 178 of the 216 samples (82%) (P = 0.019). The HBcrAg concentration correlated linearly with the HBV DNA concentration (P < 0.001) over a range which varied 100 000‐fold. The accuracy in the measurement of the patients’ HBV load obtained using the HBcrAg assay was not affected by the absence of hepatitis B e antigen from the serum or the presence of precore mutations in the HBV genome. In patients without anti‐viral drugs, changes in their serum HBcrAg concentration over time corresponded to their HBV DNA concentration. In six additional patients who were later treated with lamivudine, HBV DNA concentration declined more rapidly than their HBcrAg concentration. Three months after treatment commenced, the ratio of HBcrAg: HBV DNA had increased in all six patients (P = 0.031). The HBcrAg assay is a sensitive and useful test for the assessment of a patient's HBV load. When monitoring the anti‐viral effect of lamivudine, HBcrAg provides a viral marker which is independent of HBV DNA.
Interferon‐α combined with ketoprofen as treatment of naïve patients with chronic hepatitis C: a randomized controlled trial
Authors: Andreone P. , Gramenzi A. , Cursaro C. , Biselli M. , Lorenzini S. , Loggi E. , Felline F. , Fiorino S. , Di Giammarino L. , Porzio F. , Galli S. , Bernardi M.
Published: December 2004

Summary.  In this randomized controlled study, we evaluated the efficacy and safety of interferon‐α combined with ketoprofen to that of interferon‐α alone in naïve patients with chronic hepatitis C. Forty patients were randomized to receive Interferon‐α2a (3 million units three times a week) and ketoprofen (150 mg twice a day) and 40 to receive only interferon‐α2a at the same dose. Patients were treated for 6 months and followed up for 6 months. Response was defined by undetectable HCV‐RNA in serum at the end‐of‐treatment and after 6 months from the completion of therapy (long term response). At the end of treatment the response was similar in the two group. However, combination treatment showed significantly higher efficacy than monotherapy in achieving long term response (10%vs 32.5%; P = 0.014). Overall adverse events were similar in the two groups. ‘Flu‐like syndrome was significantly less common in the ketoprofen plus interferon group which experienced a significantly higher incidence of epigastric pain'. Our results indicate that the combination of ketoprofen plus interferon is significantly more effective than interferon alone in the tratment of naïve patients with chronic hepatitis C and is well tolerated. However this combined treatment appears to be less effective than the association of pegylated IFN and ribavirin which represent the current standard treatment. Thus, the role of ketoprofen in the treatment of chronic hepatitis C needs to be further evaluated against such a treatment.
The status of foxes Vulpes vulpes on the Isle of Man in 1999
Authors: REYNOLDS JONATHAN C. , SHORT MIKE J.
Published: December 2004

1. Foxes Vulpes vulpes probably did not occur naturally on the Isle of Man, but were present in small numbers in the mid 19th century. They were introduced again in the 1980s, and in 1990 the population was estimated at 120–300 individuals (20–50 per 100 km2) on the basis of field signs. We used a nocturnal spotlight transect survey technique to assess the status of foxes on the Isle of Man in September 1999. This method had previously been used effectively to estimate fox densities in populations as sparse as 16 per 100 km2. We surveyed a total of 852 km over a period of eight consecutive nights, during weather conditions which allowed excellent visibility. No foxes were seen.

2. By comparison with equivalent survey efforts in reference areas of mainland Britain, we conclude that post‐breeding fox density on the Isle of Man was certainly below 2.5 per 100 km2, implying a maximum of 15 foxes on the entire island. We estimate a probability of only 15–25% that a fox population of 1 per 100 km2 was present but not detected; this would be equivalent to only five or six individuals on the whole island. Foxes may even be entirely absent, although unsubstantiated sightings continue to be reported.

3. This finding is significant in understanding the ecology of the Isle of Man, and in planning the conservation of a number of ground‐nesting bird species there, as well as for farming interests, and for contingency planning against an accidental introduction of rabies.
The importance of cytotaxonomy in understanding the biogeography of African rodents: Lake Chad murids as an example
Authors: GRANJON LAURENT , DOBIGNY GAUTHIER
Published: December 2004

1. Due to the existence of numerous morphologically sibling species among African rodent genera, cytogenetical analyses are essential for correct taxonomic identifications. As an illustration, karyotypes of rodents captured in the Lake Chad area were systematically conducted, to determine them unambiguously.

2. Thirteen species were found, giving a list of at least 18 species in total for the whole Lake Chad region when previously published data were critically re‐examined. Gerbilline rodents dominate in this Sahelian region, with five species ofGerbillus, four ofTaterillus, two ofTateraandDesmodilliscus braueri, and six species of murine rodents were also counted. Moreover, two species ofTaterillusappeared characterized by unique and hitherto unknown karyotypes, and undoubtedly represent new species.

3. The Lake Chad area, although poorly studied from a biogeographical point of view, occupies a central position in the northern half of Africa which makes its zoogeographical affinities interesting to evaluate. Analysis of the biogeographical characteristics of the rodent species recorded here leads to the conclusion that the murid rodent community of the Lake Chad region is of mixed origin, with a dominant West African component. However, the presence of two species considered as endemics (Mastomys verheyeniandTaterillus lacustris), combined with our discovery of two new chromosomal species ofTaterillus, suggest that this region may have acted as a centre of differentiation in relatively recent times, perhaps reflecting the extensive variations of the Lake Chad basin associated with Plio‐Pleistocene cycles of transgression–regression.

4. Without chromosomal data, most of these findings would have remained weakly supported, or even undiscovered, as between five and eight species could have been overlooked.
The evolution of conspecific sperm precedence in Drosophila
Authors: Dixon Sheri M. , Coyne Jerry A. , Noor Mohamed A. F.
Published: December 2004

Conspecific sperm precedence takes place when females inseminated by both conspecific and heterospecific sperm preferentially produce conspecific rather than hybrid offspring. Although many studies have documented conspecific sperm precedence, most have only identified it between taxa that are already considered to be good species. Here, we test for sperm precedence between two Drosophila pseudoobscura subspecies and between two Drosophila melanogaster races to evaluate how early in evolutionary divergence sperm precedence evolves. We found evidence of weak conspecific sperm precedence between the Drosophila subspecies but none between the Drosophila races. These pairs of taxa are already separated by mating discrimination and/or hybrid sterility, so our observation suggests that conspecific sperm precedence does not always evolve before other barriers to gene exchange.
Subdivision and haplotype structure in natural populations of Arabidopsis lyrata
Authors: Wright Stephen I. , Lauga Beatrice , Charlesworth Deborah
Published: December 2004

High levels of inbreeding are expected to cause a strong reduction in levels of genetic variability, effective recombination rates and in adaptation compared with related outcrossing populations. We examined patterns of DNA polymorphism at five nuclear loci and one chloroplast locus within and between four populations of the outcrossing plant Arabidopsis lyrata, a close relative of the highly self‐fertilizing model species A. thaliana. The observed patterns are compared with species‐wide polymorphism at orthologous loci, as well as within‐ and between‐population patterns at other studied loci in A. thaliana. In addition to evidence for much higher average within‐population diversity, species‐wide levels of silent polymorphism are generally higher in A. lyrata than in A. thaliana, unlike the results from a previous study of the ADH locus. However, polymorphism is also low in the North American A. lyrata subspecies lyrata compared with the European subspecies petraea, suggesting either a population bottleneck in North American populations or recent admixture involving diverged European populations. Differentiation between the two subspecies is strong, although there are few fixed differences, suggesting that their isolation is recent. Estimates of intralocus recombination rates and analysis of haplotype structure in European A. lyrata populations indicate lower recombination than predicted based on the variability together with physical recombination rates estimated from A. thaliana. This may be due to strong population subdivision, or to recent departures from demographic equilibrium such as a bottleneck or population admixture. Alternatively, there may be consistently lower recombination rates in the outcrossing species. In contrast, estimates of recombination rates from species‐wide samples of A. thaliana are close to the values expected assuming a high rate of self‐fertilization. Complex population histories in both A. thaliana and A. lyrata complicate theoretical predictions and empirical tests of the effects of inbreeding on polymorphism and molecular evolution.
Ancestral polymorphisms in genetic markers obscure detection of evolutionarily distinct populations in the endangered Florida grasshopper sparrow (Ammodramus savannarum floridanus)
Authors: Bulgin Natalie L. , Gibbs H. Lisle , Vickery Peter , Baker Allan J.
Published: December 2004

Genetic analyses of bird subspecies designated as conservation units can address whether they represent units with independent evolutionary histories and provide insights into the evolutionary processes that determine the degree to which they are genetically distinct. Here we use mitochondrial DNA control region sequence and six microsatellite DNA loci to examine phylogeographical structure and genetic differentiation among five North American grasshopper sparrow (Ammodramus savannarum) populations representing three subspecies, including a population of the endangered Florida subspecies (A. s. floridanus). This federally listed taxon is of particular interest because it differs phenotypically from other subspecies in plumage and behaviour and has also undergone a drastic decline in population size over the past century. Despite this designation, we observed no phylogeographical structure among populations in either marker: mtDNA haplotypes and microsatellite genotypes from floridanus samples did not form clades that were phylogenetically distinct from variants found in other subspecies. However, there was low but significant differentiation between Florida and all other populations combined in both mtDNA (FST = 0.069) and in one measure of microsatellite differentiation (θ = 0.016), while the non‐Florida populations were not different from each other. Based on analyses of mtDNA variation using a coalescent‐based model, the effective sizes of these populations are large (∼80 000 females) and they have only recently diverged from each other (< 26 000 ybp). These populations are probably far from genetic equilibrium and therefore the lack of phylogenetic distinctiveness of the floridanus subspecies and minimal genetic differentiation is due most probably to retained ancestral polymorphism. Finally, levels of variation in Florida were similar to other populations supporting the idea that the drastic reduction in population size which has occurred within the last 100 years has not yet had an impact on levels of variation in floridanus. We argue that despite the lack of phylogenetic distinctiveness of floridanus genotypes the observed genetic differentiation and previously documented phenotypic differences justify continued designation of this subspecies as a protected population segment.
Genetic variation in the endangered wild apple (Malus sylvestris (L.) Mill.) in Belgium as revealed by amplified fragment length polymorphism and microsatellite markers
Authors: Coart Els , Vekemans Xavier , Smulders Marinus J. M. , Wagner Iris , Van Huylenbroeck Johan , Van Bockstaele Erik , Roldán‐Ruiz Isabel
Published: December 2004

The genetic variation within and between wild apple samples (Malus sylvestris) and cultivated apple trees was investigated with amplified fragment length polymorphisms (AFLP) and microsatellite markers to develop a conservation genetics programme for the endangered wild apple in Belgium. In total, 76 putative wild apples (originating from Belgium and Germany), six presumed hybrids and 39 cultivars were typed at 12 simple sequence repeats (SSR) and 139 amplified fragment length polymorphism (AFLP) loci. Principal co‐ordinate analysis and a model‐based clustering method classified the apples into three major gene pools: wild Malus sylvestris genotypes, edible cultivars and ornamental cultivars. All presumed hybrids and two individuals (one Belgian, one German) sampled as M. sylvestris were assigned completely to the edible cultivar gene pool, revealing that cultivated genotypes are present in the wild. However, gene flow between wild and cultivated gene pools is shown to be almost absent, with only three genotypes that showed evidence of admixture between the wild and edible cultivar gene pools. Wild apples sampled in Belgium and Germany constitute gene pools that are clearly differentiated from cultivars and although some geographical pattern of genetic differentiation among wild apple populations exists, most variation is concentrated within samples. Concordant conclusions were obtained from AFLP and SSR markers, which showed highly significant correlations in both among‐genotypes and among‐samples genetic distances.
Two centuries of the Scandinavian wolf population: patterns of genetic variability and migration during an era of dramatic decline
Authors: Flagstad Ø. , Walker C. W. , Vilà C. , Sundqvist A.‐K. , Fernholm B. , Hufthammer A. K. , Wiig Ø. , Koyola I. , Ellegren H.
Published: December 2004

The grey wolf (Canis lupus) was numerous on the Scandinavian peninsula in the early 19th century. However, as a result of intense persecution, the population declined dramatically and was virtually extinct from the peninsula by the 1960s. We examined historical patterns of genetic variability throughout the period of decline, from 1829 to 1979. Contemporary Finnish wolves, considered to be representative of a large eastern wolf population, were used for comparison. Mitochondrial DNA (mtDNA) variability among historical Scandinavian wolves was significantly lower than in Finland while Y chromosome variability was comparable between the two populations. This may suggest that long‐distance migration from the east has been male‐biased. Importantly though, as the historical population was significantly differentiated from contemporary Finnish wolves, the overall immigration rate to the Scandinavian peninsula appears to have been low. Levels of variability at autosomal microsatellite loci were high by the early 1800s but declined considerably towards the mid‐20th century. At this time, approximately 40% of the allelic diversity and 30% of the heterozygosity had been lost. After 1940, however, there is evidence of several immigration events, coinciding with episodes of marked population increase in Russian Karelia and subsequent westwards migration.