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Biomedical subjects

G Allan

Publications and source records attributed to G Allan.

At least 19 recordsLinked to original sources

PCR detection of porcine circovirus type 2 (PCV2) DNA in blood, tonsillar and faecal swabs from experimentally infected pigs.

PCV2 infection is now recognized as the major factor in the development of post-weaning multisystemic wasting syndrome (PMWS). In this study we evaluated the use of PCR to detect the presence of PCV2 DNA in blood, faecal and tonsillar swabs collected from 12 pigs experimentally infected with PCV2 and sampled at selected time points post-infection. The PCR results were evaluated together with the presence of PMWS typical histopathological lesions and the presence of PCV2 antigen. PCV2 DNA was present in the blood of all 12 infected pigs at the end of the experiment and faecal and tonsillar swabs of 11 of the 12 pigs. The rate of PCR-positive serum and plasma samples was significantly higher in four pigs that showed virological and pathological evidence of PMWS, than in infected pigs without evidence of disease. In conclusion this study confirms that PCR cannot substitute for the traditional methods used for diagnosis of PMWS, however, PCR amplification of PCV2 DNA from serum or plasma could be a useful tool to support an early diagnosis of PMWS in live animals.

Animals↗

Frequency-dependent spontaneous emission rate from CdSe and CdTe nanocrystals: influence of dark states.

We studied the rate of spontaneous emission from colloidal CdSe and CdTe nanocrystals at room temperature. The decay rate, obtained from luminescence decay curves, increases with the emission frequency in a supralinear way. This dependence is explained by the thermal occupation of dark exciton states at room temperature, giving rise to a strong attenuation of the rate of emission. The supralinear dependence is in agreement with the results of tight-binding calculations.

Journal Article↗

Rh/IGF-I/rhIGFBP-3 administration to patients with type 2 diabetes mellitus reduces insulin requirements while also lowering fasting glucose.

Administration of insulin-like growth factor-I to patients with diabetes enhances insulin action and reduces the degree of hyperglycemia but it is associated with a high rate of adverse events. Infusion of the combination of rhIGFBP-3 (the principal binding protein for IGF-I in plasma) with rhIGF-I to patients with type I diabetes improved insulin sensitivity and was associated with a low incidence in side effects. In this study, 52 patients with insulin-treated type 2 diabetes received recombinant human IGF-I plus rhIGFBP-3 in one of four dosage regimens for 14 days. The four groups were: (1) continuous subcutaneous infusion of 2 mg/kg/day; (2) the same 2 mg/kg dose infused subcutaneously over 6 h between 2000 and 0200 h; (3) 1 mg/kg twice a day by bolus subcutaneous injection; (4) a single bedtime subcutaneous injection of 1 mg/kg. Across these four groups rhIGF-I/rhIGFBP-3 decreased insulin requirements between 54% and 82%. Fasting glucose decreased by 32-37%. Mean daily blood glucose (4 determinations per day) declined in all 4 groups (range 9-23% decrease). Frequent sampling for total IGF-I, free IGF-I and IGFBP-3 was performed on days 0,1,7,14 and 15. The peak total IGF-I values were increased to 4.0-4.8-fold at 16-24 h. For free IGF-I the increase varied between 7.1 and 8.2-fold and peak values were attained at 16-20 h after administration. Both the time to maximum concentration (Tmax) and the maximum free IGF-I levels (Cmax) on day 1 for all groups were substantially less than previously published studies, wherein lower doses of rhIGF-I were given without IGFBP-3. The improvement in glucose values and the degree of reduction in insulin requirement were the greatest in groups 2 and 3 and the patients in those groups had the highest free IGF-I levels. The frequency of side effects such as edema, jaw pain and arthralgias was 4% which is less than that has been reported in previous studies wherein IGF-I was administered without IGFBP-3. We conclude that rhIGF-I/rhIGFBP-3 significantly lowers insulin requirements yet improves glucose values and these changes may reflect improvement in insulin sensitivity. Coadministration of IGFBP-3 with IGF-I produces lower free IGF-I (Tmax and Cmax) levels compared to administration of IGF-I alone and is associated with relatively low incidence of side effects during 2 weeks of administration.

Blood Glucose↗

PMWS: an emerging disease identified in archived porcine tissues.

Postweaning multisystemic wasting syndrome (PMWS) is a disease caused by porcine circovirus type 2 (PCV-2). The disease was present as early as 1986 in Spain, 1989 in Japan and 1993 in Thailand. In view of this, we considered it possible that the disease may also have been present in Switzerland prior to its first description in 2001. A retrospective investigation was performed on paraffin-embedded lymphoid organs and ileum from 496 pigs aged 5-13 weeks collected between 1976 and mid-2001. The sections were investigated immunohistochemically using a monoclonal antibody specific for PCV-2 capsid antigen encoded by ORF2. Virus antigen was detected in tissue samples of 39 pigs from 28 farms. The earliest positive sample originated from 1986. After 1989, positive pigs were found almost every year among the 20-40 cases investigated annually. These results indicate that PCV-2 has been present in Switzerland for some time, and at least since 1986.

Animals↗

Semiconducting surface reconstructions of p-type Si(100) substrates at 5 K.

We report scanning tunneling microscopy (STM) studies of the technologically important Si(100) surface that reveal at 5 K the coexistence of stable surface domains consisting of the p(2 x 1) reconstruction along with the c(4 x 2) and p(2 x 2) reconstructions. Using highly resolved tunneling spectroscopic measurements and tight binding calculations, we prove that the p(2 x 1) reconstruction is asymmetric and determine the mechanism that enables the contrast variation observed in the formation of the bias-dependent STM images for this reconstruction.

Journal Article↗

Effect of quantum confinement on the dielectric function of PbSe.

Monolayers of lead selenide nanocrystals of a few nanometers in height have been made by electrodeposition on a Au(111) substrate. These layers show a thickness-dependent dielectric function, which was determined using spectroscopic ellipsometry. The experimental results are compared with electronic structure calculations of the imaginary part of the dielectric function of PbSe nanocrystals. We demonstrate that the size-dependent variation of the dielectric function is affected by quantum confinement at well-identifiable points in the Brillouin zone, different from the position of the band-gap transition.

Journal Article↗

Features of cell degeneration and death in hepatic failure and systemic lymphoid depletion characteristic of porcine circovirus-2-associated postweaning multisystemic wasting disease.

Tissue section replicates from lymphoid tissues and livers of gnotobiotic swine were examined by immunohistochemistry for the colocalization of porcine circovirus-2 (PCV-2) nucleocapsid and terminal deoxynucleotidyl transferase (TdT)-mediated incorporation of biotinylated nucleotides (UTP) onto the 3'-exposed hydroxyl groups (nick end labeling) nuclear deoxyribonucleic acid (TUNEL), a marker for apoptosis. Single- and dually stained replicates from uninfected controls, subclinically affected PCV-2-infected gnotobiotic pigs, PCV-2-infected piglets immunosuppressed with cyclosporine (Cys), and PCV-2-infected piglets with post-weaning multisystemic wasting syndrome (PMWS) were evaluated. Thymuses were used as positive controls for apoptosis absent PCV-2, tissue sections from dogs given hyperthermic stress were examined as positive controls for induced TUNEL. Tissues from heat-stressed dogs contained TUNEL-positive cell nuclei in both lymphoid tissues and liver, TUNEL was greatest shortly after the delivery of the hyperthermic insult. In uninfected control and subclinically affected PCV-2-infected gnotobiotic pigs, rare hepatocytes and lymphoid cells were TUNEL positive, the frequency of these was similar to that seen in uninfected controls. In PMWS-affected and Cys-treated PCV-2 piglets, the only consistent strongly positive TUNEL signal was contained within the cytoplasm of virus-positive phagocytic mononuclear cells. In phagocytes, some PCV-2 inclusions were TUNEL positive. Collectively, these data indicate that apoptosis is not the primary mechanism of lymphoid depletion and hepatocyte loss in PMWS. Apoptosis associated with systemic viral diseases may be attributable to pyrexia rather than direct or indirect effects of viruses on target cells.

Animals↗

Association of lymphopenia with porcine circovirus type 2 induced postweaning multisystemic wasting syndrome (PMWS).

The composition of peripheral blood leukocyte populations was studied following experimental PCV2-infection in 3-week-old piglets. Four of 10 PCV2-infected piglets developed clinical and pathological symptoms consistent with postweaning multisystemic wasting syndrome (PMWS) between 14 and 21 days post-inoculation (p.i.), and were characterised as PMWS-affected. Only these four PMWS-affected piglets, but neither the non-symptomatic infected nor control animals, developed a clear leukopenia. Kinetic analysis demonstrated a clear loss of both CD21(+) B and CD3(+) T lymphocytes in the PMWS-affected piglets. By CD3/CD4/CD8 triple labelling, the influence of PCV2 infection on all T cell sub-populations was discernible. A loss of CD3(+)CD4(+)CD8(+) memory/activated Th lymphocytes was particularly notable. However, all T lymphocyte sub-populations-CD3(+)CD4(+)CD8(+) memory Th, CD3(+)CD4(+)CD8(-) nai;ve Th, CD3(+)CD4(-)CD8(+) Tc and CD3(+)CD4(-)CD8(-) gammadelta TCR(+) lymphocytes-were susceptible to PCV2 infection-induced lymphopenia. CD3(-)CD4(-)CD8(+) NK cells were also depleted in the PMWS-affected animals, but granulocytes and monocytes were less affected. In conclusion, PCV2 infection induces primarily a lymphopenia, but only in animals which subsequently develop PMWS. The lymphopenia can be identified early p.i., particularly with the B lymphocytes. Memory/activated Th lymphocytes might be affected more than the other T cell sub-populations, but as time progressed a collapse of both T and B cell populations was clear.

Animals↗

Optical transitions in artificial few-electron atoms strongly confined inside ZnO nanocrystals.

We have studied the optical transitions in artificial atoms consisting of one to ten electrons occupying the conduction levels in ZnO nanocrystals. We analyzed near IR absorption spectra of assemblies of weakly coupled ZnO nanocrystals for a gradually increasing electron number and found four allowed dipole transitions with oscillator strengths in quantitative agreement with tight-binding theory. Furthermore, this spectroscopy provides the single-particle energy separation between the conduction levels of the ZnO quantum dots.

Journal Article↗

Dimensionality-dependent self-energy corrections and exchange-correlation potential in semiconductor nanostructures.

We study the quasiparticle gap in semiconductor nanostructures versus dimensionality and compare it to the value obtained in the local density approximation. We first develop general arguments based on the GW approach which we then substantiate numerically by a tight binding version of this theory. We show that the gap correction is dominated by a macroscopic surface self-polarization term and point out its nonmonotonic behavior versus dimensionality.

Journal Article↗

Expression at mRNA level of cytokines and A238L gene in porcine blood-derived macrophages infected in vitro with African swine fever virus (ASFV) isolates of different virulence.

Porcine macrophage cultures were infected with two ASFV isolates of variable virulence and mRNA levels of several relevant macrophage-derived cytokines were quantified by real time PCR. At six hours post infection, a clear enhancement of mRNA expression of TNFalpha, IL6, IL12 and IL15 was observed in macrophages infected with the low virulent ASFV/NH/P68 (NHV) when compared to those infected with the highly virulent ASFV/L60 (L60). The sequence of the A238L gene homologue to the cellular IkappaB was found identical in both viral isolates and its expression at mRNA level was higher in macrophages infected with NHV when compared to macrophages infected with L60. Furthermore our results suggest a negative correlation between the mRNA expression of A238L gene and the mRNA expression of the above mentioned cytokines (with the exception of IL10) in L60 infected macrophages in opposition to the positive correlation (with exception of the IL1) suggested in NHV infection. Overall, our data strongly emphasize that virulence of ASFV isolates may depend on their capacity to regulate the expression of macrophage-derived cytokines relevant for the development of host protective responses by yet unknown mechanisms triggered by the virus at early stages of the cellular infection.

African Swine Fever↗

Methods for the exclusion of circoviruses and gammaherpesviruses from pigs.

The use of porcine tissues is being developed as a means to alleviate the shortage of allogeneic tissues and organs available for transplantation. To reduce the possibility of a microorganism of pigs being inadvertently transferred to the recipient of the xenograft, recommendations have been published on the microbiological specifications for organ source pigs. The porcine circoviruses (PCV1 and PCV2) and porcine lymphotropic herpesviruses (PLHV1 and PLHV2) are two infectious agents of pigs which are considered to be of significance for the microbiological safety of xenotransplantation. To ensure the exclusion of these microorganisms from animals destined for use under clinical conditions, reliable breeding methodologies are required. We investigated the efficiency of established derivation procedures for the removal of PCV and PLHV. In comparison with conventionally reared pigs, caesarian and barrier derived animals showed a markedly reduced prevalence of PCVs and PLHVs. Our results indicate that the derivation of animals free of both of these microorganisms is achievable and will enhance the microbiological safety of xenotransplantation.

Animals↗

Coinfection by Cryptosporidium parvum and porcine circovirus type 2 in weaned pigs.

Routine histopathological diagnosis of one representative 3-month-old pig from a group suffering from diarrhoea revealed a massive degree of parasitation by Cryptosporidium parvum, with a concomitant infection by porcine circovirus type 2 (PCV2), that was confirmed by immunohistochemical procedures. The areas of intestine where parasites were more numerous presented abundant PCV2 infected cells in mucosa and submucosa. The concurrence of C. parvum, a rare primary intestinal pathogen in post-weaning and growing pigs, and PCV2 infections suggest an increased susceptibility as a result of an immunosuppression state.

Animals↗

Reproduction of postweaning multisystemic wasting syndrome (PMWS) in immunostimulated and non-immunostimulated 3-week-old piglets experimentally infected with porcine circovirus type 2 (PCV2).

Postweaning multisystemic wasting syndrome (PMWS) in swine is causally associated with the newly recognised pathogen, porcine circovirus type 2 (PCV2). In this study, 3-week-old SPF PCV2-seronegative piglets were inoculated intranasally with PCV2. The effect of immunostimulation on the induction of PMWS was investigated by immunisation with keyhole limpet hemocyanin (KLH) emulsified in incomplete Freunds adjuvant. The study was terminated 5 weeks after inoculation. While disease was not observed in the age-matched controls, two out of five non-immunised PCV2-infected piglets died on postinoculation day (PID) 21, and one was euthanized on PID 25 in moribund condition. These animals had appeared lethargic with persistent fever from PID 12 onwards. The euthanized pig appeared smaller than littermates and suffered from jaundice. At postmortem examination, gastric ulceration, icterus, and liver and thymus atrophy were observed. Furthermore, histological lesions of degenerating hepatocytes and hepatitis in combination with lymphoid depletion and syncytial cells in lymph nodes were consistent with the diagnosis of PMWS. One out of five immunostimulated PCV2-infected piglets was euthanized on PID 22 with convulsions after a period with wasting. This pig was lethargic from PID 14 onwards with persistent fever from PID 8 and transient dyspnoea. No differences in clinical signs, gross pathologic or histological findings were observed for the remaining non-immunostimulated and immunostimulated PCV2-infected piglets. All 10 PCV2-inoculated piglets seroconverted to PCV2 within 14 days after inoculation. By virus isolation, quantitative polymerase chain reaction (Q-PCR), and immunostaining of cryostat sections, it was demonstrated that lymphoid tissue contained abundant PCV2 antigen. Viral DNA load in serum samples was assessed by Q-PCR. All four PMWS-affected piglets had high levels of PCV2 DNA in serum, suggesting that there was a correlation between high levels of viral DNA in serum and the development of PMWS. In conclusion, infection with PCV2 caused PMWS in SPF piglets, however, the immunostimulation did not seem to play a critical role.

Adjuvants, Immunologic↗

First report of post-weaning multisystemic wasting syndrome and porcine dermatitis and nephropathy syndrome in pigs in Greece.

Post-weaning multisystemic wasting syndrome (PMWS) and porcine dermatitis and nephropathy syndrome (PDNS) are two recently described conditions of pigs at the late nursery and fattening stages. The aim of this short communication was to describe the first reported occurrence of these conditions and of porcine circovirus type 2 (PCV2) infection in Greece. The clinical signs, gross post-mortem changes and histopathological changes observed in affected pigs, were similar to those previously described for both PDNS and PMWS. As in previous reports, the lesions were associated with PCV2 infection, which was demonstrated by immunohistochemical and in situ hybridization methods.

Animals↗

Depletion of CD4(+) and CD8(high+) T-cells before the onset of viraemia during classical swine fever.

Leukopenia, in particular lymphopenia, is a characteristic early event during classical swine fever (CSF). This was the case in both highly virulent (CSF virus (CSFV) strain Brescia) and moderately virulent (CSFV Uelzen) infections. The leukopenia involved leukocyte sub-populations in a disparate manner, with B-lymphocytes, helper T-cells and cytotoxic T-cells being the most affected. Depletion of lymphocyte sub-populations occurred 1-4 days before virus could be detected by RT-PCR in the serum. With the virulent Brescia virus, depletion was evident by 2 days post-infection (p.i.) but not until 3 days p.i. with an equivalent dose of the low virulent Uelzen strain. A lower (1000-fold) dose of the latter virus delayed these kinetics. gammadelta-TCR(+) T-cells were also reduced, but more so with the virulent Brescia infection. The final level of B-and alphabeta-T-cell lymphopenia was similar for all animals, including those infected with the lower virus dose. AnnexinV staining revealed that cell viability was clearly diminished, particularly interesting, considering the clinical differences between infections by Brescia and Uelzen viruses. It was the time p.i. and rate of appearance of dying cells which was more rapid in the virulent Brescia infections. Interestingly, the repeated blood sampling resulted in depletion of some leukocyte populations also in non-infected control animals. Particularly neutrophils and NK cells, and to a lower extent CD4(+), CD8(+) T-lymphocytes and B-lymphocytes were affected. Taken together, the data show that the alphabeta-T-lymphocyte subsets are particularly susceptible to modulation during the acute phase of CSF, being detectable before the onset of viraemia. The pathogenic mechanism therein would involve indirect virus-host interactions, probably originating from the site of primary infection, rather than a direct effect of the virus or viral protein. Furthermore, these characteristics offer an explanation for the retardation of the cellular and humoral immune response observed during classical swine fever.

Animals↗

Activation of the immune system is the pivotal event in the production of wasting disease in pigs infected with porcine circovirus-2 (PCV-2).

Porcine circovirus (PCV)-2, a newly described single-stranded circular DNA virus pathogen of swine is the cause of postweaning multisystemic wasting syndrome (PMWS). In gnotobiotic piglets, PCV-2 infection alone produces asymptomatic infection without evidence of overt PMWS. Gnotobiotic piglets infected with PCV-2 were injected with keyhole limpet hemocyanin in incomplete Freund's adjuvant (KLH/ICFA), and the effects on virus production and development of PMWS were determined. In the first experiment, piglets were injected subcutaneously on the left hip and shoulder, and viral burden was assessed in regional lymph nodes draining the injection sites and in contralateral lymph nodes 13-14 days after infection. Immune activation increased the number of virus antigen-positive cells in draining lymph nodes and increased the amount of infectious virus recovered by 1-4 log10. In a second experiment, the effects of injections of KLH/ICFA with or without concurrent stimulation of peritoneal macrophages by intraperitoneal injections of thioglycollate broth on induction of PMWS was assessed. All immunized piglets developed moderate to severe PMWS, whereas none of the piglets infected with PCV-2 alone developed PMWS. In PMWS-affected piglets, extensive replication of PCV-2 was documented by both immunocytochemistry and quantitative viral titrations. Thus, immune activation is a key component of the pathogenesis of PCV-2-associated PMWS in swine.

Adjuvants, Immunologic↗