bluuebunny/crossref_metadata_2025 huggingface dataset:
Relationship between the incidence of type 1 diabetes and enterovirus infections in different European populations: Results from the EPIVIR project
Authors: Viskari Hanna , Ludvigsson Johnny , Uibo Raivo , Salur Lina , Marciulionyte Dalia , Hermann Robert , Soltesz Gyula , Füchtenbusch Martin , Ziegler Anette‐G. , Kondrashova Anita , Romanov Anatolij , Knip Mikael , Hyöty Heikki
Published: February 2004

The incidence of type 1 diabetes varies markedly between countries. As enterovirus infections have been linked to type 1 diabetes, we determined whether this variation correlates with the frequency of enterovirus infections in different Caucasian populations in Europe. Enterovirus antibodies were examined in the background population (1‐year‐old and 10‐14–year‐old children) in seven countries with either exceptionally high (Finland and Sweden) or low/intermediate incidence of diabetes (Estonia, Germany, Hungary, Lithuania, Russia) using EIA and neutralisation assays. Enterovirus antibodies were less frequent in countries with high diabetes incidence compared to countries with low diabetes incidence (P < 0.001). This suggests that enterovirus infections are not particularly common in countries with high diabetes incidence. In contrast, there seems to be an inverse correlation between the incidence of type 1 diabetes and enterovirus infections in the background population, which is in line with the previously proposed polio hypothesis according to which the complications of enterovirus infections become more common in an environment with a decreased rate of infections. J. Med. Virol. 72:610–617, 2004. © 2004 Wiley‐Liss, Inc.
Differential display RT‐PCR analysis of ECV304 endothelial‐like cells infected with dengue virus type 2 reveals messenger RNA expression profiles of multiple human genes involved in known and novel roles
Authors: Liew Kingsley J.L. , Chow Vincent T.K.
Published: February 2004

Differential display (DD)‐RT‐PCR was employed to analyze mRNAs from ECV304 human endothelial‐like cells undergoing apoptosis following infection with the virulent New Guinea C strain of dengue virus type 2 in order to elucidate the cellular gene responses to dengue viral infection at the transcriptional level. We isolated, sequenced, and identified 203 differentially expressed and overlapping cDNA fragments, all of which were of human origin except 1 that was of viral origin. Out of these, 78 were individual distinct clones comprising 46 and 32 expressed sequence tags (ESTs) that exhibited upregulated and downregulated trends, respectively. Of the 78 differentially expressed mRNAs, 16 did not match any characterized genes or ESTs. The remaining 62 mRNAs modified by dengue virus infection matched known genes, including those encoding components of the cell cycle (Anillin, CDC27), cytoskeleton (ε‐tubulin), signal transduction (OPHN1, PPP2R2A, TIRAP), protein translation and modification (EIF3S10, IF2, TMEM1), transcriptional regulation (α‐NAC, C20orf104, EGR1, ELP2), apoptotic cell death (RICK), membrane (BPAG1), and mitochondrial‐related proteins. Semiquantitative RT‐PCR and real‐time RT‐PCR authenticated further the altered expression patterns of selected genes of interest. These data demonstrate the feasibility of mRNA DD in providing insights into the complex responses of the transcriptional machinery of permissive and apoptotic human endothelial‐like cells in the pathogenesis of dengue and/or its complications induced by the virulent dengue virus type 2. J. Med. Virol. 72:597–609, 2004. © 2004 Wiley‐Liss, Inc.
A novel CD4+ T‐helper lymphocyte epitope in the VP3 protein of hepatitis A virus
Authors: Sánchez Glòria , Pintó Rosa M. , Bosch Albert
Published: February 2004

Prediction analysis of TH‐cell epitopes in the VP3 capsid protein of the hepatitis A virus (HAV) revealed the occurrence of a putative TH epitope in the 102–121 region complying with all the algorithms tested. To confirm these predictions, spleen T lymphocytes obtained from BALB/c mice previously immunised with HAV, were stimulated in vitro with different concentrations of synthetic peptides 102–121 and 110–121 of VP3. The ability of these peptides to stimulate CD4+ T‐helper lymphocytes proliferation was evaluated by an immunological flow cytometry detection of bromodeoxyuridine incorporation. Using this method, it is concluded that the amino terminal part of the VP3 102–121 sequence contains a T cell epitope. J. Med. Virol. 72:525–532, 2004. © 2004 Wiley‐Liss, Inc.
Genetic variability of hepatitis C virus non‐structural protein 3 and virus‐specific CD8+ response in patients with chronic hepatitis C
Authors: López‐Labrador F. Xavier , He Xiao‐Song , Berenguer Marina , Cheung Ramsey C. , González‐Candelas Fernando , Wright Teresa L. , Greenberg Harry B.
Published: February 2004

Hepatitis C virus (HCV) variation in specific T‐cell epitopes may represent a mechanism of viral persistence in chronic infection. We examined the HCV non‐structural protein 3 (NS3), including the immunologically relevant epitopes HCV NS3‐2 KLVALGINAV (human leukocyte antigen [HLA]‐A2‐restricted) and HCV NS3‐1391 LIFCHSKKK (HLA‐A3‐restricted), in 22 HLA‐A2+ patients with chronic infection. Significant amino acid variation was found in HCV NS3‐2 epitope sequences when compared to the HCV‐1 prototype virus. Six of the nine different HCV NS3‐2 peptide variants were identified in patients with HCV NS3‐2‐specific CD8+ cells, detected with an HLA‐A2 tetramer made with the HCV‐1 prototype peptide. Phylogenetic analysis, including HCV reference sequences other than HCV‐1, suggested however that most of the variations in the HCV NS3‐2 epitope could be related to genetic heterogeneity between HCV reference subtypes. Variation was less common when comparing HCV NS3‐2 epitope sequences from the clinical isolates to the most‐closely related HCV reference subtype in each case. Some subtype‐independent variations were found in epitopic residues probably important for T‐cell receptor interaction. In contrast, no significant variation was found in HLA primary anchor sites, flanking regions, or in the contiguous HLA A3‐restricted CD8+ T‐cell epitope. Ongoing variation was not evident in two selected patients with follow‐up. In conclusion, (i) the HCV NS3‐2 epitope is not conserved between different HCV strains/subtypes, and (ii) an HLA‐A2 tetramer loaded with the HCV‐1 prototype NS3‐2 peptide may still detect NS3‐specific CD8+ cells in some patients with variant viruses. These data may be useful to improve T‐cell assays using HCV NS3 peptides, taking into account the genetic diversity of this virus. J. Med. Virol. 72:575–585, 2004. © 2004 Wiley‐Liss, Inc.
Association of JC virus with tubulointerstitial nephritis in a renal allograft recipient
Authors: Wen Mei‐Chin , Wang Chia‐Liang , Wang Meilin , Cheng Chi‐Hung , Wu Ming‐Ju , Chen Cheng‐Hsu , Shu Kuo‐Hsiung , Chang Deching
Published: February 2004

A renal allograft transplant patient with tubulointerstitial nephritis was examined for possible involvement of human polyomaviruses. Biopsy of the allograft kidney revealed inflammatory interstitial infiltration with tubulitis and typical inclusion bodies in the nucleus of the tubulo‐epithelium. Glomeruli showed duplication and lamination of Bowman's capsule with enlargement of the parietal and visceral epithelia. Immunohistochemical analysis on the graft section using a JC virus (JCV) VP1 peptide‐specific antibody showed positive staining. Paracrystalline arrays of naked viral particles with diameters of 40 nm were visualized in the nuclear inclusions under an electron microscope. Molecular examination using the polymerase chain reaction (PCR) and DNA sequencing revealed that the JCV was involved in the nephritis. The results indicated that JCV may be associated with tubulointerstitial nephritis. J. Med. Virol. 72:675–678, 2004. © 2004 Wiley‐Liss, Inc.
Epidemiological and antigenic analysis of respiratory syncytial virus in hospitalised Tunisian children, From 2000 to 2002
Authors: Fodha I. , Vabret A. , Trabelsi A. , Freymuth F.
Published: February 2004

Human respiratory syncytial virus (RSV) is the major viral cause of lower respiratory tract disease in children. Infections with the virus occur as annual winter epidemics in temperate climates, placing considerable pressure on the provision of hospital beds. Most molecular epidemiological studies have, until now, focused on isolates from infants in industrialised countries. No data have been available with regard to RSV strains from northern Africa.In this report, a recent RSV outbreak in Tunisia was studied and results showed that 176 of 815 (21.6%) nasopharyngeal aspirates collected from hospitalised children were RSV‐positive by immunofluorescence assay. This RSV outbreak showed a temperature‐dependant pattern (P = 0.026) but no significant association with rainfall. A total of 73 RSV‐positive samples were tested by two reverse‐transcription‐polymerase chain reaction assays (RT‐PCR): RT‐PCR‐1, which amplifies the RNA of all RSV strains, and RT‐PCR‐2, which allows subgroup classification of RSV. Analysis by hybridisation assay of RT‐PCR‐2‐amplified 1B protein gene products showed a higher prevalence of group B RSV than that of group A (82.5% vs. 17.5% among the typed strains). Knowledge of the variants is important in terms of both diagnosis and definition of a vaccine composition. J. Med. Virol. 72:683–687, 2004. © 2004 Wiley‐Liss, Inc.
Analysis of B‐lymphocyte differentiation in patients infected with hepatitis C virus
Authors: Fournillier Anne , Freida Delphine , Defrance Thierry , Merle Philippe , Trépo Christian , Inchauspé Geneviève
Published: February 2004

To clarify whether some of the functions of B lymphocytes could be affected during hepatitis C virus (HCV) infection, phenotypic characteristics of B lymphocytes from HCV‐infected patients and their capacity to differentiate into immunoglobulins (Ig)‐secreting cells were studied. B lymphocytes differentiation was investigated for patients untreated and non‐responders (n = 9), treated and non‐responders (n = 6), responders (n = 6), long‐term responders (n = 9) to therapy and seronegative controls (n = 14) following in vitro stimulation with S. aureus strain Cowan I mitogen. HCV sequences in purified B lymphocytes were detected by RT‐PCR. It was found that HCV‐patients harbor a similar mean percentage of B cells and a normal level of naïve B cells (% IgM+/IgD+ cells = 79.7 ± 15.4 for untreated non‐responders, 57.1 ± 22.9 for treated non‐responders, 44.3 ± 29.1 for responders, 75.7 ± 16 for long‐term responders) as compared with controls. It was also found that peripheral blood mononuclear cells (PBMCs) of patients or controls produced similar amounts of IgG, A, and M in vitro. A total of 57% of untreated non‐responders versus 17% of treated non‐responders were able to produce HCV‐specific antibodies. Interestingly, B lymphocytes from PBMCs able to secrete anti‐HCV antibodies contained HCV positive strand RNA, although no systematic detection of the negative strand was found. These data suggest that signaling through the B cell receptor (BCR) in B lymphocytes of HCV‐infected patients appears normal whatever their response to therapy. The capacity to secrete HCV‐specific IgG seemed to be linked to the presence of positive strand RNA rather than virus replication. J. Med. Virol. 72:566–574, 2004. © 2004 Wiley‐Liss, Inc.
Epidemiology of hepatitis A and hepatitis E infection and their determinants in adult Chinese community in Hong Kong
Authors: Wong K.H. , Liu Y.M. , Ng P.S.P. , Young B.W.Y. , Lee S.S.
Published: February 2004

Current epidemiology of hepatitis A virus (HAV) and hepatitis E virus (HEV) in Hong Kong was evaluated in 936 adult Chinese subjects recruited through a telephone interview in 2001. Some 15% of the subjects had IgG antibodies to both HAV and HEV while 665 (71.0%) and 176 (18.8%) had anti‐HAV and anti‐HEV, respectively. Age was the most significant independent factor. Six hundred thirty‐eight (79.8%) and 165 (20.7%) subjects aged ≥30 had anti‐HAV and anti‐HEV, respectively, as compared with 27 (19.7%) and 11 (8.0%) in people aged <30. The corresponding adjusted Odds ratio (OR) was 14.94 (95% CI: 9.13–24.44; P < 0.001) for anti‐HAV positivity and 2.99 (95% CI: 1.58–5.67; P = 0.001) for anti‐HEV positivity. Subjects born outside Hong Kong were more likely to have anti‐HAV (adjusted OR: 3.41; 95% CI: 2.21–5.26; P < 0.001) but not anti‐HEV. Non‐labour work people were less likely to have anti‐HAV‐adjusted OR, 0.40 (95% CI: 0.26–0.62; P < 0.001). Age‐specific HAV prevalence right shifted in the last 20 years. Anti‐HAV positivity was less frequent, across all age groups, in subjects >21‐years‐old in the present study than another study done in 1987–89 (P < 0.001). HAV prevalence only increased slightly in every 10‐year age groups of people aged 21–50 when compared with their corresponding 10‐year‐younger age groups (P = 0.11), suggesting an ageing cohort effect with no major infections in the last decade. For HEV, both the overall and age‐specific prevalence decreased over the last decade (P < 0.001). The increasing proportion of susceptible population to enterically transmitted viral hepatitis has implications to future prevention and control programmes, including vaccination strategies. J. Med. Virol. 72:538–544, 2004. © 2004 Wiley‐Liss, Inc.
Microarray technology for identification and distinction of hantaviruses
Authors: Nordström Henrik , Johansson Patrik , Li Quan‐Gen , Lundkvist Åke , Nilsson Peter , Elgh Fredrik
Published: February 2004

DNA microarrays combine high‐precision technology with advanced molecular biology to achieve high‐throughput screening of DNA fragments. In this study, we investigated the potential of the cDNA microarray technique to identify and discriminate PCR derived amplicons from genetically highly similar viruses. The wide range of sequence variation among hantaviruses makes them suitable as a model for this purpose. The hantaviruses, carried by rodents, cause several hundred thousand cases of severe human disease every year in many parts of the world. A hantavirus‐specific microarray, including DNA fragments from 12 viral isolates of six different hantaviruses, was designed. The S and M genome segments were represented by 500‐nucleotide overlapping and 250‐nucleotide non‐overlapping fragments. A considerable ability to distinguish between different hantaviruses was demonstrated using a novel analysis method. Even different isolates of a single virus, were identified correctly despite 90% sequence similarity. The distinction ability was accompanied by a tolerance for smaller sequence differences, which makes the microarray suitable for testing samples containing unknown viruses. Viral genetic material found in samples from the lungs of bank voles caught in the wild was identified precisely, which demonstrated further the potential for this technology. J. Med. Virol. 72:646–655, 2004. © 2004 Wiley‐Liss, Inc.
High seroprevalence of human herpesvirus 8 (HHV‐8) in HIV‐1‐infected pregnant women of Southeastern Italy: Association with injection drug use and hepatitis C virus infection
Authors: Fiore Josè Ramòn , Volpe Anna , Tosatti Maria Alessandra , Valentin Lucia De , Favia Anna , Chironna Maria , Lisco Andrea , Vimercati Antonella , Angarano Gioacchino , Chieco‐Bianchi Luigi , Calabrò Maria Luisa
Published: February 2004

The seroprevalence of human herpesvirus 8 (HHV‐8) in a group of HIV‐1‐infected pregnant women and in mother–child pairs from Southeastern Italy (Apulia) was determined. Blood was collected from 49 HIV‐1‐infected women during pregnancy or at delivery as well as from their children. Samples were analysed for the presence of antibodies to the latency‐associated nuclear antigen and a structural antigen encoded by open reading frame 65. The presence of antibodies to hepatitis C virus (HCV) was also determined. Nineteen women (38.7%) were found to be positive for HHV‐8 antibodies to at least one of the two antigens, and 21 (42.9%) for HCV antibodies. HHV‐8 antibodies were more common in injecting drug users (56.3%) than in women infected through heterosexual intercourse (30.3%). HCV antibodies were significantly more prevalent in HHV‐8‐seropositive (66.7%) than HHV‐ 8‐seronegative (29%) women. Thirteen children born to HIV‐1/HHV‐8 co‐infected women were HHV‐8‐seroreactive, with a variable pattern of reactivity to the analysed antigens. Follow‐up of children showed a prolonged persistence of antibodies, in two cases for more than 12 months. This study has provided serological evidence for a high rate of HHV‐8 infection in HIV‐1‐infected women in the Apulia region, and has identified a possible association between HHV‐8 infection, past use of injection drugs and HCV infection. Parenteral transmission may, therefore, be a mode of virus spread. J. Med. Virol. 72:656–660, 2004. © 2004 Wiley‐Liss, Inc.
Mechanisms of coxsackievirus B5 mediated β‐cell death depend on the multiplicity of infection
Authors: Rasilainen Suvi , Ylipaasto Petri , Roivainen Merja , Lapatto Risto , Hovi Tapani , Otonkoski Timo
Published: February 2004

Coxsackievirus infections may trigger and accelerate pancreatic β‐cell death, leading to type I diabetes. Unrestricted coxsackievirus B5 replication in cultured β‐cells inoculated with high multiplicity leads to rapid lytic cell death. Evidence from other virus‐host cell systems indicates that host cell responses to infection may depend on the multiplicity of infection (MOI). Thus, the aim of this study was to compare the mechanisms of β‐cell death during high versus low multiplicity of coxsackievirus B5 infection. Cultures of highly differentiated mouse insulinoma cells and primary adult human islets were infected with coxsackievirus B5 at multiplicities of >1,000 or <0.5 TCID50 per cell. The results of nuclear morphology and viability stainings, TUNEL staining and electrophoretic DNA fragmentation analysis showed high multiplicity infection to predominantly induce necrosis and transient apoptosis. In low multiplicity culture, however, necrosis was only moderately induced and apoptosis increased steadily with time. This was best demonstrated by a tenfold higher apoptosis/necrosis ratio than after high multiplicity inoculation. Expression of γ‐glutamyl cysteine synthetase increased in both infective cultures but the level of intracellular glutathione permanently depleted only at high multiplicity and recovered fully at low multiplicity. Thus, apoptosis represents an important mechanism of β‐cell death after low multiplicity of coxsackievirus B5 infection. This process is associated with maintenance of a physiological intracellular glutathione profile differing dramatically from the high multiplicity infection during which necrosis dominates and intracellular thiol balance deteriorates. These data suggest that the pattern and mechanisms of coxsackievirus B5 infection induced β‐cell death depend on the MOI. J. Med. Virol. 72:586–596, 2004. © 2004 Wiley‐Liss, Inc.
Comparison of genotypes C and D of the hepatitis B virus in Japan: A clinical and molecular biological study
Authors: Duong Tran Nhu , Horiike Norio , Michitaka Kojiro , Yan Chen , Mizokami Masashi , Tanaka Yasuhito , Jyoko Kouji , Yamamoto Kazutoshi , Miyaoka Hiroaki , Yamashita Yoshimasa , Ohno Naofumi , Onji Morikazu
Published: February 2004

The genotype‐related differences between genotype C and genotype D of the hepatitis B virus (HBV) remain unknown. The relationship was studied between the HBV genotypes and their clinical features, paying special attention to genotypes C and D. Serum samples from 413 HBV carriers were genotyped using an enzyme immunoassay (EIA) and by restriction fragment length polymorphism. The nucleotide sequences at the basic core promoter (BCP) and precore (PreC) regions were analysed by direct sequencing. The full genome sequences of three HBV genotype D cases were also examined. Almost all carriers with HBV genotype D were asymptomatic carriers (84.2%). Genotype D was not found in patients with liver cirrhosis and hepatocellular carcinoma. In contrast, carriers with genotype C had mainly chronic liver disease (63.2%; P < 0.001). The ratio of hepatitis B e antigen (HBeAg)/anti‐HBe was significantly higher in genotype C than in genotype D in the young age‐matched group (P < 0.01). The mutation at BCP (T1762, A1764) was significantly lower in genotype D than in genotype C among HBeAg‐negative patients (P < 0.05). The HBV full‐genome sequences are very similar to certain HBV genotype D sequences from Europe. In conclusion, genotype C was associated with chronic liver disease, whereas genotype D was related to asymptomatic carriers with earlier HBeAg seroconversion. Thus, the outcome of chronic HBV infection may be different in persons infected with HBV genotypes C and D. J. Med. Virol. 72:551–557, 2004. © 2004 Wiley‐Liss, Inc.
Pneumonia in HIV‐infected patients in the HAART era: Incidence, risk, and impact of the pneumococcal vaccination
Authors: López‐Palomo C. , Martín‐Zamorano M. , Benítez E. , Fernández‐Gutiérrez C. , Guerrero F. , Rodríguez‐Iglesias M. , Girón‐González J.A.
Published: February 2004

The objective of this study was to assess the factors implicated in an increased or decreased risk of pneumonia, with particular attention to the response to highly active antiretroviral therapy (HAART) and the effect of the polysaccharide 23‐valent pneumococcal vaccination in 300 human immunodeficiency virus (HIV)‐infected adults followed‐up for a median of 35.6 months. Pneumococcal pneumonia occurred in 12 patients and all bacterial pneumonia (pneumonia caused by Streptococcus pneumoniae or other bacteria, as well as those with negative cultures but presumably bacterial in origin) in 40 patients. In the univariate analysis, immunodepressed patients (defined as those with less than 200 CD4+ T cell/μl), those without immunological response to HAART (defined as an increase of 25% of CD4+ T lymphocyte count), patients with previous admissions to hospital and those with cotrimoxazole or Mycobacterium avium intracellulare prophylaxis showed a higher incidence of both pneumococcal and all bacterial pneumonia. Multivariate analysis demonstrated that the presence of pneumococcal pneumonia was associated with a CD4+ lymphocyte count at the time of HIV diagnosis <200 cells/μl. The multivariate model that was more valid for prediction of all bacterial pneumonia included a CD4+ T cell count <200 cells/μl and absence of immunological response to HAART. Only in patients with a baseline CD4+ T cell count lower than 200/μl and immunological response to HAART, a near significant lower incidence of all bacterial pneumonia was observed after vaccination. Thus, these results do not support an important additional protective effect of 23‐valent pneumococcal vaccine in HIV‐patients with immunological response to HAART. J. Med. Virol. 72:517–524, 2004. © 2004 Wiley‐Liss, Inc.
Role of complement in neutralization of respiratory syncytial virus
Authors: Yoder Sandra M. , Zhu Yuwei , Ikizler Mine R. , Wright Peter F.
Published: February 2004

Serum neutralizing antibody titers to respiratory syncytial virus (RSV) are higher when assayed with guinea pig complement. A number of different mechanisms have been suggested for enhancement of neutralization by complement. The most straightforward is that complement–antibody complexes present a greater steric hindrance to viral entry than with antibody alone. To define the implications of measuring serum neutralizing antibody with and without complement, sera from adults, young children, infants, and cord bloods were run in plaque neutralization assays with representative viruses of the RSV A and B subgroups. Although titers of neutralizing antibody were higher in the presence of complement, the addition of complement did not increase the ability to detect antibody rises after natural infection. Some of the complement effect may be attributable to an inhibition of RSV replication by complement alone. While these observations do not address the role for complement in the pathogenesis of RSV infection, they suggest that neutralization assays performed without complement may be most reflective of physiologic conditions in the respiratory tract. J. Med. Virol. 72:688–694, 2004. © 2004 Wiley‐Liss, Inc.
Reversal of toxigenic effects of aflatoxin B1on cockerels by alcoholic extract of African nutmeg,Monodora myristica
Authors: Oluwafemi Flora , Taiwo Victor O
Published: February 2004

The reversal of the toxigenic effects of crystalline aflatoxin B1(AFB1; fromAspergilus flavus) on cockerels using different concentrations of the ethanolic extract from the seeds of African nutmeg,Monodora myristica, was studied in 130 two‐week‐old cockerels, randomly divided into 13 groups (A–M) of ten birds each. Group A (controls) received 100 µl of phosphate‐buffered saline, while those in the test groups B, C and D received 690 ng, 1380 ng or 2010 ng of AFB1, respectively; groups E to M received the three doses of AFB1with either 0.1, 0.2 or 0.3 ml of the extract ofM myristica, respectively, 5 min after administration of AFB1. Jugular blood, collected on days 14 and 21 after administration, was analysed for haematology parameters, serum proteins, alanine aminotransaminase (ALT), aspartate aminotransaminase (AST) and γ‐glutamyl transferase (GGT) levels. The birds were observed daily for clinical signs and mortality for 21 days. Post‐mortem examination was carried out on the livers of dead birds and those sacrificed by exsanguination after the 21 days post‐administration. One hundred per cent mortality was recorded in cockerels that received high doses of only AFB1within 72 h (2010 ng per bird), and between 17 and 19 days (1380 ng). The cockerels developed leucocytosis and microcytic hypochromic anaemia. Dosing with 0.2 or 0.3 ml of the extract ofM myristicareversed these changes over time, while the lowest dose (0.1 ml) did not. While serum bilirubin levels rose significantly in test cockerels, no significant changes (P> 0.05) were observed in the levels of serum proteins of control and test cockerels, whether or not they received the extract ofM myristica. Aflatoxicosis caused significant decreases and increases (P< 0.05) in serum AST and GGT levels, respectively, while the administration of alcoholic extract ofM myristicareversed the GGT but not AST levels to normal. Livers from control cockerels and those given 2010 ng of AFB1showed no appreciable gross or histological lesions. The other groups showed mild to moderate hepatomegaly, paleness and friability, diffuse vacuolar degeneration and necrosis, Kupffer cell proliferation, presence of megalocytes and multinucleated hepatocytes. Cockerels that received 0.2 or 0.3 ml of extract ofM myristica, irrespective of the dose of AFB1showed the mildest hepatic lesions. The results of this study show that AFB1is highly toxigenic to cockerels and that this could be successfully reversed by the concurrent administration of alcoholic extract ofM myristica. It is therefore recommended that reduction of fungal growth on foods and subsequent toxicosis by aeration, cooling, modified atmospheres or by fungistats, is complemented with the use of extracts of spices fromM myristicaseed, a tree widely distributed along the west coast and central Africa. Copyright © 2004 Society of Chemical Industry
Copper bioavailability in rats fed unused palm olein and palm olein used in frying
Authors: Pérez‐Granados Ana M , Vaquero M Pilar , Navarro M Pilar
Published: February 2004

The influence of the consumption of unused and used (in repeated potato frying operations) palm olein on copper bioavailability was investigated. Three diets containing respectively 8% olive oil (O), palm olein (P) and palm olein from 80 repeated potato frying operation without turnover (PF), which nearly reached the limit of 25% polar compounds allowed by law, were given to rats for 28 days. No differences were found in food intake and body weight. The liver mass index was higher in PF than in P rats. No significant differences were observed in copper apparent absorption or in liver, skin, erythrocyte, serum and carcass copper contents. Therefore consumption of palm olein, whether unused or used in frying, does not alter copper bioavailability in growing rats. Copyright © 2004 Society of Chemical Industry
Changes in carotenoids and chlorophylls in fresh green asparagus (Asparagus officinalis L) stored under modified atmosphere packaging
Authors: Tenorio MD , Villanueva MJ , Sagardoy M
Published: February 2004

Asparagus is a highly perishable product owing to its high respiratory rate, a reason why modified atmosphere packaging (MAP) has been used in order to increase the shelf‐life of fresh asparagus. Carotenoid and chlorophyll pigments are important compounds for the maintenance of both the nutritional and sensory quality of asparagus. In this study, green asparagus spears were stored under refrigeration at 2 °C for 14 days, under MAP at 2 °C for 26–33 days and under MAP at 10 °C for 20 days. Pigment profiles were determined using high‐performance liquid chromatography (HPLC), with three classes of compounds being detected, namely oxygenated carotenoids (xanthophylls), hydrocarbonated carotenoids (carotenes) and chlorophylls. The xanthophylls identified were neoxanthin, violaxanthin, lutein and zeaxanthin. Only β‐carotene was detected in the carotene fraction. In the chlorophyll pigments, three molecules were isolated, chlorophyll b, its epimer chlorophyll b′ and chlorophyll a. Also detected were the first degradation products of chlorophylls, pheophytins b and a respectively. Modified atmosphere packaging at 2 °C was found to be effective in extending the shelf‐life for up to 4 weeks and in preserving the colour of fresh asparagus. Copyright © 2004 Society of Chemical Industry
Bunker‐stored silage covered with biodegradable coating. Part I. Laboratory assay
Authors: Denoncourt Patrick , Caillet Stéphane , Lacroix Monique
Published: February 2004

The effect of a hydrophobic biodegradable coating on the conservation of silage was evaluated in comparison with unsealed silage and silage covered with a polyethylene (plastic). Silage samples were subjected to biochemical analysis (pH, dry matter, water‐soluble sugars, ammoniacal nitrogen, lactic acid) and microbiological analysis (total aerobes, lactic acid bacteria, moulds and yeasts) during 135 days of storage. Silage covered with the biodegradable coating had a constant pH value of 4.79 and was preserved against spoilage until day 120, whereas the pH of unsealed silage increased during the first 6 days to a value of 8.22. After 2 months of storage, silage covered with the biodegradable coating had a lower dry matter content, higher water‐soluble sugar content and higher lactic acid concentration than unsealed silage. During storage the population of total aerobes was lower in silage covered with the biodegradable coating than in silage covered with plastic. Furthermore, silage covered with the biodegradable coating had a stable content of aerobic micro‐organisms. The biodegradable‐coated silage showed a growth of lactic acid bacteria similar to that of silage covered with plastic. Only silage covered with the biodegradable coating had a low initial content of moulds and yeasts. Air infiltration was present throughout the storage period in the plastic and biodegradable coating treatments and was linked with the degradation of silage in both cases. However, the biodegradable coating significantly (P ≤ 0.05) increased the period of silage conservation compared with that of uncoated silage. Copyright © 2004 Society of Chemical Industry
Vitamin E levels, thiobarbituric acid test and sensory evaluation of breast muscles from broilers fed α‐tocopheryl acetate‐ and β‐carotene‐supplemented diets
Authors: Carreras Irene , Guerrero Luis , Guàrdia MD , Esteve‐Garcia Enric , García Regueiro JA , Sárraga Carmen
Published: February 2004

The objective of this study was to investigate the effect of dietary α‐tocopheryl acetate and β‐carotene supplementation on lipid oxidation of breast meat from broilers fed lard as the fat source. Supplementation of broilers with 100 mg kg−1 α‐tocopheryl acetate increased the vitamin E levels in raw breast samples significantly (p < 0.05), whereas the presence of 1.5 mg kg−1 dietary β‐carotene tended to decrease vitamin E deposition. The presence of vitamin E delayed lipid oxidation significantly, but thiobarbituric acid values of samples from broilers fed the β‐carotene‐supplemented diet did not differ from those of control samples. Vitamin E reduced sensory meat rancidity, whilst vitamin E, β‐carotene and their combination modified meat texture. The results show the effectiveness of dietary α‐tocopheryl acetate supplementation in protecting broiler meat against lipid oxidation. Copyright © 2004 Society of Chemical Industry
The Cannizzaro‐like metabolites of secoiridoid glucosides in some olive cultivars
Authors: Piperno Anna , Toscano Marirosa , Uccella Nicola A
Published: February 2004

Oleuropein (the most abundant bitter principle) and its analogue ligstroside, both secoiridoid biophenols, were extracted from Hojiblanca black olives and analysed by preparative flash chromatography and HPLC. During olive growth, maturation and processing, these compounds undergo biological and technological degradation to various metabolites, by enzymatic degradation through esterases, phenoloxidases and β‐glucosidases. An enzymatic system with regiospecific Cannizzaro‐like activity could be responsible for two novel metabolites, isolated and characterised by 1H‐ NMR, 12C NMR and FAB‐MS as hydroxycarboxylic and lactone derivatives of terpenoid origin from the precursor oleoside‐11‐methylester, formed from both secoiridoid biophenols. The occurrence of these two secoiridoid metabolites requires a thermodynamically controlled sequential process, via hydride rearrangements, through the complex metabolic pathway. The possible reaction sequence was interpreted by molecular calculations performed on competitive isomers. Copyright © 2004 Society of Chemical Industry
Condensed tannins in humid tropical fodder crops and their in vitro biological activity: Part 1
Authors: López Juan , Tejada Irma , Vásquez Carlos , Garza Juan de Dios , Shimada Armando
Published: February 2004

A descriptive study was conducted in order to determine the tannin profiles of tropical fodder crops. The following types of condensed tannins (CT) were determined (expressed as g kg−1 of dry matter): free (FCT), bound to protein (CTP), bound to fibre (CTF) and total (TCT). High FCT levels were found in Desmodium ovalifolium (228.4), Flemigia macrophylla (181.9), Guazuma ulmyfolia (129.7) and Leucaena leucocephala (129.5). The highest concentrations of CTP were detected in Gliricidia sepium (61.0), F macrophylla (23.3), and G ulmyfolia (21.1). CTF levels were the highest in G ulmyfolia (55.1), F macrophylla (46.7) and G sepium (41.5). Analyzed samples cultivated in vertisol and fluvisol soils showed different contents of tannins. It has been reported that plants with more than 60 g kg−1 FCT are less palatable and digestible than forages with lower concentrations of this chemical class, although there is more protein to bypass the rumen and higher nitrogen retention. Graminaceous plants showed higher concentrations of FCT and TCT concentrations were generally lower than legumes and other forage crops. Copyright © 2004 Society of Chemical Industry
Examination of the sulfur dioxide–ascorbic acid anti‐oxidant system in a model white wine matrix
Authors: Bradshaw Mark P , Scollary Geoffrey R , Prenzler Paul D
Published: February 2004

The effectiveness of sulfur dioxide in inhibiting the ascorbic acid induced oxidation of (+)‐catechin in a matrix simulating white wine was investigated. At a sulfur dioxide:ascorbic acid mole ratio of 0.8:1.0, sulfur dioxide extended the delay prior to the onset of (+)‐catechin browning as measured by the increase in absorbance at 440 nm from 2 to 4 days post reaction initiation. The lengthening of the pre‐browning period was correlated with an increase in the time required to oxidise ascorbic acid completely, the sequence of events being loss of sulfur dioxide (day 3), loss of ascorbic acid (day 4) and on‐set of (+)‐catechin browning (day 4). Increasing the sulfur dioxide to ascorbic acid mole ratio to 3.0:1.0 inhibited the onset of (+)‐catechin browning over the 14‐day reaction period. This inhibition was achieved at considerable cost to the sulfur dioxide concentration, with the ratio of sulfur dioxide consumed to ascorbic acid oxidised being 1.7 compared with the expected fraction of 1.0. Reasons for the enhanced consumption of sulfur dioxide and the implications of this study for maintenance of quality in white wine production are discussed. Copyright © 2004 Society of Chemical Industry
Condensed tannins in tropical fodder crops and their in vitro biological activity: Part 2
Authors: López Juan , Tejada Irma , Vásquez Carlos , Garza Juan de Dios , Shimada Armando
Published: February 2004

With the aim to evaluate the biological activity of purified condensed tannins of tropical forages we conducted two in vitro experiments. In the first, using a radial diffusion technique, the protein precipitation of free condensed tannins (FCT) from Manihot esculenta, leucaena leucocephala, Arachis pintoi, Guazuma ulmyfolia, Gliricidia sepium and of tannic acid on bovine serum albumin (ASB), papain, pepsin and trypsin at pH 5.0 and 6.8 was evaluated with a three‐way analysis of variance. Significant effects (P ≤ 0.05) for the tannin type, protein source, pH and their interactions were observed. Pepsin showed the highest protein precipitation (PP) at a pH of 5.0 (82.9 µg) with FCT of G ulmyfolia and the lowest (0 and 0.2 µg) of BSA with G sepium and A pintoi at pH 6.8. Experiments were then conducted using completely randomized designs in order to observe the effect of adding 0, 1.25 or 2.50 mg of FCT from M esculenta and L leucocephala to the rumen fluid‐buffer in an in vitro dry matter digestibility test (IVDMD) of Medicago sativa and Brachiaria decumbens, The IVDMD value of M sativa (757 g kg−1) decreased with L leucocephala tannins, although with those of M esculenta it was increased (824 g kg−1 and 871 g kg−1, respectively) for 1.25 and 2.5 mg of FCT. The IVDMD value of B decumbens (774 g kg−1 without tannins) diminished with any tannin and any dose (P < 0.05). We conclude that there are differences in the FCT contents of fodder crops and in their biological activity measured as the capacity to precipitate proteins, which is modified by the type of tannin, the protein and the pH. The result of an IVDMD is regulated by the type of tannin and its dose. Copyright © 2004 Society of Chemical Industry
Kinetics of peptide fraction release during in vitro digestion of casein
Authors: Agudelo Rafael A , Gauthier Sylvie F , Pouliot Yves , Marin Johanne , Savoie Laurent
Published: February 2004

The charges and sizes of peptides released during in vitro casein digestion were investigated. Casein was hydrolysed sequentially by pepsin and pancreatin and, during hydrolysis, the products of digestion were removed by dialysis. The undigested residues were separated by ion‐exchange chromatography into basic–neutral (BN), lightly acid (LA) and acid (A) fractions. Each of these fractions was further resolved by sequential ultrafiltration (MWCO 10 and 1 kDa) into two retentates (>10 and 10–1 kDa) and one permeate (<1 kDa). The polypeptides (>10 kDa) produced by pepsin hydrolysis were degraded into small molecules during the first 2 h of pancreatin hydrolysis; less than 100 mg g−1 of the total N remained undigested. Most of the material produced was in the BN10‐1 (ie basic–neutral, 10–1 kDa) fraction, with three times as much N as the A10‐1 fractions. As digestion progressed a decrease in the proportion of N in the residues retained by the dialysis membrane was observed. This decrease was particularly rapid in the BN10‐1 fraction. Large proportions of leucine (Leu), lysine (Lys), arginine (Arg), phenylalanine (Phe) and tyrosine (Tyr) were found as peptides smaller than 1 kDa, both in the dialysates and retentates, while glutamine (Glu), threonine (Thr), serine (Ser) and asparagine (Asp) appeared mostly in the A10‐1 and A > 10 fractions. After 6 h of pancreatin hydrolysis most of the proline (Pro) content was in the BN10‐1 fractions. The mechanisms behind and the implications of these results are discussed. Copyright © 2004 Society of Chemical Industry
Irradiation of maize and bean flours: effects on starch physicochemical properties
Authors: Rombo George O , Taylor John RN , Minnaar Amanda
Published: February 2004

In an earlier study (Rombo GO et al, J Sci Food Agric 81: 497–502 (2001)) it was found that the in vitro starch digestibility of raw and cooked maize and bean flours was increased by irradiation of the flours at 2.5 kGy, but there was a reduction in starch digestibility at doses above 2.5 kGy. Experiments were performed to determine what effects irradiation had on the molecular properties of the starch in maize and bean flours. Increasing irradiation dose caused an increased proportion of β(1–3)‐ and β(1–4)‐bonded starch in bean and maize flours. Starch containing β‐bonds is only partially digestible by porcine pancreatic α‐amylase, and this may in part explain the reduction in starch digestibility at higher doses. Size exclusion–high‐performance liquid chromatography showed that higher irradiation doses led to a reduction in the molecular size of amylopectin in both bean and maize starches, which presumably involved debranching and an increase in the production of short, straight‐chain molecules. With increased irradiation dose there was an increase in the crystallinity of the amylopectin fraction of the bean starch as shown by differential scanning calorimetry, presumably at least in part due to β‐bonding and amylopectin depolymerisation. These two factors and their interrelated effects are probably responsible for the observed slight reduction in maize and bean porridge starch digestibility reported previously. Copyright © 2004 Society of Chemical Industry
Furosine is a useful indicator in pre‐baked breads
Authors: Cardenas Ruiz Jonathan , Guerra‐Hernandez Eduardo , García‐Villanova Belén
Published: February 2004

There is increasing demand for pre‐baked bread. Bakers can meet consumer requirements for breads of specific flavour and colour by pre‐baking an amount of bread in the morning and completing the baking process in the afternoon. This type of product is of special interest to sandwich bars, restaurants and large communities. In order to obtain an indicator of utility to monitor the processing of pre‐baked bread, the browning indicators furosine, hydroxymethylfurfural (HMF) and glucosylisomaltol were analysed in two independent assays of bread pre‐baked at 175 °C for between 5 and 30 min. No furosine was detected in raw or fermented dough. In the pre‐baking stage, furosine increased from 1.5‐ to 5‐fold between 5 and 30 min. The furosine values obtained in the two independent experiments were similar. HMF and glucosylisomaltol were only detected after 15 min, which is the time period commonly used for processing by the industry. Thus, among these indicators, furosine is the only one with utility for monitoring the pre‐baking process. On the other hand, a study of the baking of pre‐baked bread at 220 °C showed that HMF and glucosylisomaltol can be used to monitor the latter process. Copyright © 2004 Society of Chemical Industry
Effect of gluten content on recrystallisation kinetics and water mobility in wheat starch gels
Authors: Wang Xin , Choi Sung‐Gil , Kerr William L
Published: February 2004

The effect of gluten on starch retrogradation at 5 °C was studied using 1H NMR relaxometry. Gels were made from gluten and starch at 27.8 and 38.5% total solids and with gluten comprising either 10, 15 or 20% of the solids. Changes in the transverse relaxation time constant (T2) were related to water mobility. Mono‐exponential analysis of relaxation curves showed that, in general, gluten retarded starch retrogradation. T2 values in gluten gels also decreased during storage, but to a much lesser extent. Distributed exponential analysis showed that two distinct regions of T2 were observed in all samples. During aging, the peak values of both regions shifted to lower values for all gels. Starch gel samples showed the most significant shift, and gluten gels showed the least. The three levels of gluten addition in starch/gluten gels produced similar shifts. For all samples the signal intensity of the less mobile region decreased more dramatically than that of the more mobile region during storage. It was suggested that gluten retards water loss in the granule remnants. Copyright © 2004 Society of Chemical Industry
Colour characteristics of honeys as influenced by pollen grain content: a multivariate study
Authors: Terrab Anass , Escudero M Luisa , González‐Miret M Lourdes , Heredia Francisco J
Published: February 2004

A chromatic analysis by tristimulus colorimetry and a pollen analysis (pollen grains contained in each honey sample, considering their volume and geometrical shape) were carried out on 33 Eucalyptus unifloral honeys; the colour of the pollen grains was also considered. Multiple linear regression (MLR) and partial least squares regression (PLSR) were used to establish equations relating the chromatic variables to the pollen data, ie number and morphology of the pollen grains, thus allowing the prediction of the ultimate colour from the botanical characteristics. The results obtained show that lightness (L*) is significantly (p < 0.05) related to the pollen type Olea europaea; on the other hand, the variable that better relates to the chroma $(C_{ab}^{*})$ is the Zea mays pollen type. Copyright © 2004 Society of Chemical Industry
Technical note on a much simplified method for collecting ruminal fluid using a nylon paint strainer
Authors: Leyendecker Sandra A , Callaway Todd R , Anderson Robin C , Nisbet David J
Published: February 2004

Collection of ruminal fluid for in vitro experimentation has traditionally been accomplished by straining rumen digesta through multiple layers of cheesecloth or other cumbersome filtering materials, such as surgical gauze, nylon tissues or wire gauzes. We here report a comparison of the gross physical, fermentative and microbial characteristics of ruminal fluid collected by straining through cheesecloth or through a nylon mesh paint strainer that, because of its pouched shape, is conveniently filled with digesta thus making the collection and straining process much easier. Whether strained through cheesecloth or the nylon mesh strainer, ruminal fluid did not differ (P > 0.05) in dry matter, neutral or acid detergent fiber content or in concentrations of total culturable anaerobes. Total amounts (mM) of volatile fatty acid or ammonia produced after 24 h of incubation of cheesecloth‐ or nylon mesh‐strained ruminal fluid with added tryptose, SigmaCell 50 or starch did not differ (P > 0.05) thus indicating that the microbial populations within the strained fluids possessed similar abilities to ferment these test substrates. Because the physical, fermentative and microbial characteristics of the nylon mesh‐ and cheesecloth‐strained ruminal fluid were essentially equivalent, we conclude that the much easier to use nylon mesh strainers can be incorporated into a faster, more convenient ruminal fluid straining method. Published in 2004 for SCI by John Wiley & Sons, Ltd.