bluuebunny/crossref_metadata_2025_split huggingface dataset:
Expression of Werner and Bloom syndrome genes is differentially regulated by in vitro HIV-1 infection of peripheral blood mononuclear cells
Authors: BORDI L , AMENDOLA A , CICCOSANTI F , ABBATE I , CAMILLONI G , CAPOBIANCHI M R 
Published: October 2004

SUMMARY
In HIV infection, continuous immune activation leads to accelerated ageing of the adaptive immune system, similar to that observed in elderly people. We investigated the expression of WRN and BLM (genes involved in disorders characterized by premature ageing, genomic instability and cancer predisposition) in peripheral blood mononuclear cells (PBMC) activated in vitro with phytohaemagglutinin (PHA) and infected with different HIV-1 strains. The steady state levels of mRNA were analysed by reverse transcription-polymerase chain reaction (RT-PCR), and protein expression was assayed using immunocytochemistry and Western blot techniques. In uninfected PBMC, PHA stimulation induced an increase in BLM mRNA and protein expression, while WRN expression remained virtually unchanged. When PBMC were infected in vitro with a lymphotropic HIV-1 strain, the level of BLM mRNA showed a peak at 24 h of infection, followed by a decline to uninfected culture levels. A similar result failed to be seen using an R5-tropic HIV-1 strain. In accordance with mRNA expression, in HIV-infected cultures PBMC were stained more frequently and more intensely by a BLM-specific antibody as compared to uninfected cultures, staining peaking at 24. Conversely, WRN expression was not modulated by HIV-1. The proportion of cells showing BLM up-regulation, established by immunocytochemical staining, was much greater than the proportion of productively infected PBMC, as established by proviral DNA measurement. This result indicates that BLM up-regulation is probably a result of an indirect bystander cell effect. Activation of the BLM gene in infected PBMC suggests that premature ageing could be a further immunopathogenetic mechanism involved in HIV-induced immunodeficiency, and points to a possible new candidate target for innovative therapeutic intervention.
Variable NKG2 expression in the peripheral blood lymphocytes of rhesus monkeys
Authors: LABONTE M L , LETVIN N L 
Published: October 2004

SUMMARYTo provide a basis for beginning to explore the CD94/NKG2 family of molecules in rhesus monkeys, we sought to characterize the expression of these inhibitory and activating cell signalling molecules in peripheral blood mononuclear cells (PBMCs) from healthy rhesus monkeys. We developed and employed a semiquantitative polymerase chain reaction (PCR)-based assay to evaluate mRNA expression levels of nine NKG2 molecules in PBMCs from the monkeys. In addition to quantitating NKG2A, NKG2B, NKG2C2, NKG2C and NKG2D expression, mRNA expression of transmembrane-deleted forms of these molecules was also evaluated. Significant variability in NKG2 mRNA expression in the PBMCs was detected, with 15 unique NKG2 expression level profiles detected in a study of 15 monkeys. We also found that the ratio of the expressed levels of mRNA of the four NKG2 splice variants, NKG2A, NKG2B, NKG2AΔtm, and NKG2BΔtm, was variable between the monkeys as well as in an individual monkey over a period of 1·5 years. These findings indicate the dynamic nature of NKG2 mRNA expression in the rhesus monkey.
Association of increased platelet-associated immunoglobulins with thrombocytopenia and the severity of disease in secondary dengue virus infections
Authors: SAITO M , OISHI K , INOUE S , DIMAANO E M , ALERA M T P , ROBLES A M P , ESTRELLA B D , KUMATORI A , MOJI K , ALONZO M T , BUERANO C C , MATIAS R R , MORITA K , NATIVIDAD F F , NAGATAKE T 
Published: October 2004

SUMMARYSevere thrombocytopenia and increased vascular permeability are two major characteristics of dengue haemorrhagic fever (DHF). To develop a better understanding of the roles of platelet-associated IgG (PAIgG) and IgM (PAIgM) in inducing thrombocytopenia and its severity of disease in patients with secondary dengue virus infection, the relationship between the PAIgG or PAIgM levels and disease severity as well as thrombocytopenia was examined in 78 patients with acute phase secondary infection in a prospective hospital-based study. The decrease in platelet count during the acute phase recovered significantly during the convalescent phase. In contrast, the increased levels of PAIgG or PAIgM that occurred during the acute phase of these patients decreased significantly during the convalescent phase. An inverse correlation between platelet count and PAIgG or PAIgM levels was found in these patients. Anti-dengue virus IgG and IgM activity was found in platelet eluates from 10 patients in an acute phase of secondary infection. Increased levels of PAIgG or PAIgM were significantly higher in DHF than those in dengue fever (DF). An increased level of PAIgM was associated independently with the development of DHF, representing a possible predictor of DHF with a high specificity. Our present data suggest that platelet-associated immunoglobulins involving antidengue virus activity play a pivotal role in the induction of thrombocytopenia and the severity of the disease in secondary dengue virus infections.
Heterogeneous expression of apolipoprotein‐E by human macrophages
Authors: Tedla Nicodemus , Glaros Elias N. , Brunk Ulf T. , Jessup Wendy , Garner Brett 
Published: October 2004

SummaryApolipoprotein‐E (apoE) is expressed at high levels by macrophages. In addition to its role in lipid transport, macrophage‐derived apoE plays an important role in immunoregulation. Previous studies have identified macrophage subpopulations that differ substantially in their ability to synthesize specific cytokines and enzymes, however, potential heterogeneous macrophage apoE expression has not been studied. Here we examined apoE expression in human THP‐1 macrophages and monocyte‐derived macrophages (MDM). Using immunocytochemistry and flow cytometry methods we reveal a striking heterogeneity in macrophage apoE expression in both cell types. In phorbol‐ester‐differentiated THP‐1 macrophages, 5% of the cells over‐expressed apoE at levels more than 50‐fold higher than the rest of the population. ApoE over‐expressing THP‐1 macrophages contained condensed/fragmented nuclei and increased levels of activated caspase‐3 indicating induction of apoptosis. In MDM, 3–5% of the cells also highly over‐expressed apoE, up to 50‐fold higher than the rest of the population; however, this was not associated with obvious nuclear alterations. The apoE over‐expressing MDM were larger, more granular, and more autofluorescent than the majority of cells and they contained numerous vesicle‐like structures that appeared to be coated by apoE. Flow cytometry experiments indicated that the apoE over‐expressing subpopulation of MDM were positive for CD14, CD11b/Mac‐1 and CD68. These observations suggest that specific macrophage subpopulations may be important for apoE‐mediated immunoregulation and clearly indicate that subpopulation heterogeneity should be taken into account when investigating macrophage apoE expression.
Characterization of the B‐cell inhibitory protein factor in Ixodes ricinus tick saliva: a potential role in enhanced Borrelia burgdoferi transmission
Authors: Hannier Sigrid , Liversidge Janet , Sternberg Jeremy M. , Bowman Alan S. 
Published: October 2004

SummaryWe recently described the inhibition of host B lymphocytes by Ixodes ricinus tick saliva. In this study, we characterized the factor responsible for this activity and examined the modulation of lipopolysaccharide (LPS)‐ and Borrelia burgdorferi outer surface protein (Osp)‐induced proliferation of naive murine B lymphocytes by an enriched fraction of this factor. The B‐lymphocyte inhibitory activity was destroyed by trypsin treatment, indicating that a proteinaceous factor was responsible for this activity. The removal of glutathione‐S‐transferase (GST) from tick salivary glands extracts (SGE) showed that this B‐cell inhibitory protein (BIP) was not a GST. Gel filtration liquid chromatography indicated that BIP has a native molecular weight of ≈ 18 000. An enrichment protocol, using a combination of anion‐exchange and reverse‐phase liquid chromatography, was established. BIP‐enriched fractions did not suppress T‐cell proliferation. Delayed addition of BIP‐enriched fractions, up to 7 hr after LPS addition, inhibited the proliferation of isolated B cells. BIP‐enriched fractions dramatically inhibited both OspA‐ and OspC‐induced proliferation of isolated B cells. These results strongly suggest that BIP may facilitate B. burgdorferi transmission by preventing B‐cell activation, and also highlights the potential of BIP as a therapeutic agent in B‐cell maladies.
A novel model of sensitization and oral tolerance to peanut protein
Authors: Strid Jessica , Thomson Melanie , Hourihane Jonathan , Kimber Ian , Strobel Stephan 
Published: October 2004

SummaryThe prevalence of food allergic diseases is rising and poses an increasing clinical problem. Peanut allergy affects around 1% of the population and is a common food allergy associated with severe clinical manifestations. The exact route of primary sensitization is unknown although the gastrointestinal immune system is likely to play an important role. Exposure of the gastrointestinal tract to soluble antigens normally leads to a state of antigen‐specific systemic hyporesponsiveness (oral tolerance). A deviation from this process is thought to be responsible for food‐allergic diseases. In this study, we have developed a murine model to investigate immunoregulatory processes after ingestion of peanut protein and compared this to a model of oral tolerance to chicken egg ovalbumin (OVA). We demonstrate that oral tolerance induction is highly dose dependent and differs for the allergenic proteins peanut and OVA. Tolerance to peanut requires a significantly higher oral dose than tolerance to OVA. Low doses of peanut are more likely to induce oral sensitization and increased production of interleukin‐4 and specific immunoglobulin E upon challenge. When tolerance is induced both T helper 1 and 2 responses are suppressed. These results show that oral tolerance to peanut can be induced experimentally but that peanut proteins have a potent sensitizing effect. This model can now be used to define regulatory mechanisms following oral exposure to allergenic proteins on local, mucosal and systemic immunity and to investigate the immunomodulating effects of non‐oral routes of allergen exposure on the development of allergic sensitization to peanut and other food allergens.
Immunoassays: their history, development and current place in food science and technology
Authors: Bonwick Graham A. , Smith Christopher J. 
Published: October 2004

AbstractThe purpose of this paper is to introduce the reader to immunoassays. This paper is the first in a themed issue of the Journal in which a number of papers have been brought together in order to demonstrate the types and variety of immunoassays, which are currently available. Indeed it might be said that all an analyst needs to do is to name a molecule and somewhere there will now be an immunoassay for the detection of that molecule. This obviously is not entirely accurate, however immunoassays do provide a powerful tool, which can be used in the analysis and quality control of food materials. For both the novice and the experienced worker the specialist terminology of a subject presents an initial barrier, which must be overcome before full understanding is achieved. In this paper an attempt is made to introduce the important terms with which the reader should be familiar and to try to set the various technologies in context. The various basic methods are described and the theoretical and practical basis of more sophisticated assays now being devised are introduced.
Fluorescence polarization immunoassay based on a monoclonal antibody for the detection of ochratoxin A
Authors: Shim Won‐Bo , Kolosova Anna Yu. , Kim Yoon‐Jung , Yang Zheng‐You , Park Seon‐Ja , Eremin Sergei A. , Lee In‐Seon , Chung Duck‐Hwa 
Published: October 2004

SummaryA fluorescence polarization immunoassay (FPIA) based on a monoclonal antibody for the determination of ochratoxin A (OTA) was developed. Fluorescein‐labelled OTA derivative (tracer) was synthesized and purified by thin‐layer chromatography. The optimized OTA FPIA had a dynamic range from 5 to 200 ng mL−1 with IC50 value of 30 ng mL−1 and a detection limit of 3 ng mL−1. The method developed was characterized by high specificity and reproducibility. Cross‐reactivity with other mycotoxins (zearalenone, aflatoxins, patulin and T‐2 toxin) was negligible (<0.1%). Methanol extracts of barley samples were used for the analysis. The results of OTA determination in barley were compared with those determined by indirect competitive enzyme‐linked immunosorbent assay (ELISA). Recoveries for the samples spiked at 50, 100 and 500 ng g−1 levels were 91, 90 and 97%, respectively, for FPIA, and 98, 98 and 102%, for ELISA. Naturally contaminated barley samples were analysed by these methods but some disagreement was observed between the results. The FPIA method can be applied for screening of food samples for OTA residues without a complicated clean‐up.
Species identification of meat products by ELISA
Authors: Giovannacci Inès , Guizard Cécile , Carlier Martine , Duval Valérie , Martin Jean‐Luc , Demeulemester Claude 
Published: October 2004

SummaryELISA methods used in this study are proved to detect low contents of animal species (pork, beef, sheep and poultry), even in highly processed foods. They present the advantages of being robust, cheap and easy to perform. Nevertheless, F factors, determining the threshold values of the test, need to be validated for each species.
Application of immunological methods for the detection of species adulteration in dairy products
Authors: Hurley Ian P. , Elyse Ireland H. , Coleman Robert C. , Williams John H. H. 
Published: October 2004

SummaryA number of enzyme‐linked immunosorbent assays (ELISAs) have been developed for the detection of milk adulteration in dairy products. Target antigens have been caseins, lactoglobulins, immunoglobulins and other whey proteins. Polyclonal and monoclonal antibodies have been used in a variety of formats including direct, indirect, competitive and sandwich ELISAs. ELISAs have been successfully applied to the detection of cows' milk adulteration of sheep, goat and buffalo milk. Goat milk adulteration of sheep milk has also been detected. A number of ELISAs have also been applied to cheese. It is recommended that ELISA should be used in combination with PCR to ensure compliance with current legislation.
Mortality in patients with diabetic neuropathic osteoarthropathy (Charcot foot)
Authors: Gazis A. , Pound N. , Macfarlane R. , Treece K. , Game F. , Jeffcoate W. 
Published: October 2004

AbstractObjective  To determine the mortality of a population of patients diagnosed with Charcot neuropathic osteoarthropathy managed by a single specialist unit and to compare the results with a control population.Methods  We have undertaken a retrospective analysis of all cases of Charcot foot on the comprehensive database which has been maintained at the specialist diabetic foot clinic at the City Hospital, Nottingham since 1982. Survival and the incidence of amputation (major and minor) was compared with a control population referred with uncomplicated neuropathic ulceration. Controls were individually matched for gender, age (±2 years), disease type, disease duration (±2 years) and year of referral (±3 years).Results  Forty‐seven cases (21 female, 26 male) of Charcot foot were identified, of whom 18 (38.3%) had Type 1 diabetes. Mean age and disease duration at presentation were 59.2 ± 13.4 (sd) and 16.2 ± 11.2 years, compared with 59.7 ± 12.6 and 16.3 ± 11.2 years, respectively, in the controls. Twenty‐one (44.7%) of those with Charcot had died, after a mean interval of 3.7 ± 2.8 years. This compared with 16 (34.0%) after a mean 3.1 ± 2.7 years in the control group. Mean duration of follow‐up in the survivors was 4.7 ± 4.9 years (Charcot) and 5.3 ± 3.9 years (controls). A total of 11 (23.4%) Charcot patients had had a major amputation on the side of the index lesion, compared with five (10.6%) controls. There was no difference between the two groups (P > 0.05, Chi‐square).Conclusions  The mortality in this group of patients with Charcot foot was higher than expected. Nevertheless, there was no difference between those with Charcot and those with uncomplicated neuropathic ulceration. It is possible that it is neuropathy, rather than Charcot osteoarthropathy, which is independently associated with increased mortality in diabetes. The mechanism underlying any such association is not known. There is a need for a formal, prospective, multicentre study to investigate the life expectancy and cardiovascular risk of those with Charcot osteoarthropathy.
Does autonomic neuropathy play a role in erythropoietin regulation in non‐proteinuric Type 2 diabetic patients?
Authors: Spallone V. , Maiello M. R. , Kurukulasuriya N. , Barini A. , Lovecchio M. , Tartaglione R. , Mennuni G. , Menzinger G. 
Published: October 2004

AbstractAims  Erythropoietin (EPO)‐deficient anaemia has been described in Type 1 diabetic patients with both severe autonomic neuropathy (AN) and proteinuria. This study was aimed at distinguishing between the effects of AN and nephropathy on haemoglobin and EPO levels in Type 2 diabetic patients at an early stage of diabetic nephropathy.Methods  In 64 Type 2 diabetic patients (age 52 ± 10 years, duration 10 ± 9 years) without overt nephropathy and other causes of anaemia or EPO deficit, we assessed cardiovascular tests of AN, 24‐h blood pressure (BP) monitoring, urinary albumin excretion rate (UAE), a full blood count, and serum EPO.Results  Although the Type 2 diabetic patients with AN did not show differences in haemoglobin and EPO when compared with patients without AN, the presence of haemoglobin < 13 g/dl was associated with the presence of AN (χ2= 3.9, P < 0.05) and of postural hypotension (χ2= 7.8, P < 0.05). In a multiple regression analysis including as independent variables gender, body mass index, duration of diabetes, smoking, creatinine, 24‐h UAE, 24‐h diastolic BP, ferritin, erythrocyte sedimentation rate, and autonomic score, we found that the only variables independently related to haematocrit were autonomic score, ferritin and erythrocyte sedimentation rate. Finally, the physiological inverse relationship between EPO and haemoglobin present in a control group of 42 non‐diabetic non‐anaemic subjects was completely lost in Type 2 diabetic patients. The slopes of the regression lines between EPO and haemoglobin of the control subjects and the Type 2 diabetic patients were significantly different (t = 14.4, P < 0.0001).Conclusions  This study documents an early abnormality of EPO regulation in Type 2 diabetes before clinical nephropathy and points to a contributory role of AN in EPO dysregulation.
A prospective study of Type 2 diabetes and depressive symptoms in the elderly: The Rancho Bernardo Study
Authors: Palinkas L. A. , Lee P. P. , Barrett‐Connor E. 
Published: October 2004

AbstractAims  The association between Type 2 diabetes and depressive symptoms was examined prospectively to assess possible causal relationships between the two diseases.Methods  A cohort of 971 men and women aged 50 and older from the adult population of Rancho Bernardo, California had an oral glucose tolerance test and completed the Beck Depression Inventory (BDI) at two clinic visits, 1984–87 and 1992–96.Results  Depressive symptoms at baseline were associated with higher follow‐up levels of non‐fasting plasma glucose (P = 0.001) and an increased risk of developing Type 2 diabetes [odds ratio (OR) = 2.50; 95% confidence interval (CI) = 1.29–4.87], independent of sex, age, exercise and body mass index. Conversely, baseline non‐fasting plasma glucose was not significantly associated with follow‐up depressive symptoms and Type 2 diabetes at baseline was not significantly associated with the onset of BDI scores ≥ 11 by the second visit (OR = 0.73; 95% CI = 0.41–1.30).Conclusions  Depressed mood is more likely to be a risk factor for Type 2 diabetes in older adults than the reverse.
Long‐term renoprotection by perindopril or nifedipine in non‐hypertensive patients with Type 2 diabetes and microalbuminuria
Authors: Jerums G. , Allen T. J. , Campbell D. J. , Cooper M. E. , Gilbert R. E. , Hammond J. J. , O'Brien R. C. , Raffaele J. , Tsalamandris C. ,   The Melbourne Diabetic Nephropathy Study Group
Published: October 2004

AbstractAims  To assess the efficacy of an angiotensin converting enzyme (ACE) inhibitor (perindopril), a dihydropyridine calcium channel blocker (sustained release nifedipine) and placebo in preventing the progression of albuminuria and decline in glomerular filtration rate (GFR) in patients with Type 2 diabetes and microalbuminaria.Methods  A prospective, randomized, open, blinded end point study of 77 patients allocated to three treatment groups (23 perindopril, 27 nifedipine, 27 placebo). Drug doses were adjusted to achieve a decrease in diastolic blood pressure (DBP) of 5 mmHg in the first 3 months and additional therapy was given if hypertension developed (supine DBP > 90 mmHg and/or systolic blood pressure (SBP) > 140 mmHg if ≤ 40 years; supine DBP > 90 mmHg and/or SBP > 160 mmHg if > 40 years). Median follow‐up was 66 months, with 37 patients being followed for at least 6 years.Results  Blood pressure remained within the non‐hypertensive range in 83% of perindopril‐, 95% of nifedipine‐ and 30% of placebo‐treated patients (P < 0.01). In the first 12 months albumin excretion rate (AER) decreased by 47% only in the perindopril group (P = 0.04). From 12 to 72 months, AER gradients increased by 27% per year only in the placebo group (P < 0.01). After 6 years, macroalbuminuria had developed in 7/15 placebo compared with 2/11 in perindopril and 1/11 nifedipine‐treated patients (P = 0.05). GFR did not change in the first 12 months, but thereafter the median GFR gradient (ml/min/1.73 m2 per year) was −2.4 (P < 0.01) for perindopril‐, −1.3 (P = 0.26) for nifedipine‐ and −4.2 (P = 0.01) for placebo‐treated patients. The rate of decline in GFR for the study group as a whole from 12 months to the end of follow‐up correlated negatively with mean arterial pressure (MAP) (r = −0.38, P < 0.01). During a 3‐month treatment pause in 29 patients AER tended to increase only in the perindopril group (P < 0.07).Conclusions  Long‐term control of blood pressure with perindopril or nifedipine stabilizes AER and attenuates GFR decline in proportion to MAP in non‐hypertensive patients with Type 2 diabetes and microalbuminuria.
Suppression of the biosynthesis of proanthocyanidin in Arabidopsis by a chimeric PAP1 repressor
Authors: Matsui Kyoko , Tanaka Hideo , Ohme‐Takagi Masaru 
Published: October 2004

SummaryFlavonoids are secondary metabolites that are specific to higher plants. PAP1, a member of the family of MYB domain transcription factors in Arabidopsis, is a positive regulator of the biosynthesis of anthocyanin. We show here that a chimeric PAP1 repressor, in which the EAR‐motif repression domain from SUPERMAN was fused to PAP1, suppressed the expression of four flavonoid biosynthetic genes, namely CHS, DFR, LDOX, and BAN, in siliques, and inhibited the accumulation of proanthocyanidin, even in the presence of an endogenous positive regulator, such as TT2. This suppression resulted in the formation of light yellow seeds in 35S::PAP1SRDX transgenic plants. Our results indicate that PAP1 has the potential ability to regulate the biosynthesis not only of anthocyanin but also of proanthocyanidin. Our gene silencing system, using chimeric repressors, appears to be a useful tool for the manipulation of the biosynthesis of secondary metabolites in plants.