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E Sinclair

Publications and source records attributed to E Sinclair.

At least 19 recordsLinked to original sources
PubMed2001

Use of overlapping peptide mixtures as antigens for cytokine flow cytometry.

Intracellular cytokine staining and flow cytometry can be used to measure T-cell responses to defined antigens. Although CD8+ T-cell responses to soluble proteins are inefficiently detected by this approach, peptides can be used as antigens. Using overlapping peptides spanning an entire protein sequence, CD8+ T-cell responses can be detected to multiple epitopes, regardless of HLA type. In this study, overlapping peptide mixes of various lengths were compared and 15 amino acid peptides with 11 amino acid overlaps were found to stimulate both CD4+ and CD8+ T-cell responses. Such peptide mixes stimulated CD4+ T-cell responses equivalent to those observed with whole recombinant protein, while simultaneously stimulating CD8+ T-cell responses much higher than those observed with whole protein. Although 8-12 amino acid peptides produced the highest level of CD8+ T-cell responses, 15 amino acid peptides were still very effective. Peptides that were 20 amino acids in length, however, did not stimulate strong CD8+ T-cell responses at the same peptide dose. The cytokine responses to individual epitopes added up approximately to the response to the entire mix, demonstrating that large mixes can detect responses in a quantitative fashion. Unlike whole protein antigens, peptide mixes were effective at stimulating responses in both cryopreserved PBMC and blood stored for 24 h at room temperature. Thus, overlapping 15 amino acid peptide mixes may facilitate the analysis of antigen-specific CD4+ and CD8+ T-cell responses by cytokine flow cytometry, using clinical specimens that include shipped blood or cryopreserved PBMC.

Clinical Trials as Topic↗
PubMed2001

Training programs in school consultation.

The need to train psychiatrists in school consultation is important to approach mental health and psychosocial concerns from the broad perspective of addressing barriers to learning and promoting healthy development. There is a major national impetus to improve academic achievement and literacy, which can be amplified by addressing the social, emotional, and mental health needs of children and youth. Training in school consultation allows the psychiatrist to better understand a critical institution in each child's life and also provides technical assistance and training to school personnel, which facilitates networking between programs and individuals involved in or interested in school mental health. Each of the described programs approaches consultation from a specific focus that varies in time commitment, placement options, and frequency and order of didactic presentations. There is no conclusive available evidence as to which program is most effective, since such evaluation depends on the overall goals of the consultation program itself.

Adolescent↗
PubMed1997

The nef gene products of both simian and human immunodeficiency viruses enhance virus infectivity and are functionally interchangeable.

Adult rhesus macaques infected with nef-defective simian immunodeficiency virus (SIV) exhibit extremely low levels of steady-state virus replication, do not succumb to immunodeficiency disease, and are protected from experimental challenge with pathogenic isolates of SIV. Similarly, rare humans found to be infected with nef-defective human immunodeficiency virus type 1 (HIV-1) variants display exceptionally low viral burdens and do not show evidence of disease progression after many years of infection. HIV-1 Nef induces the rapid endocytosis and lysosomal degradation of cell surface CD4 and enhances virus infectivity in primary human T cells and macrophages. Although expression of SIV Nef also leads to down-modulation of cell surface CD4 levels, no evidence for SIV Nef-induced enhancement of virus infectivity was observed in earlier studies. Thus, it remains unclear whether fundamental differences exist between the activities of HIV-1 and SIV Nef. To establish more clearly whether the SIV and HIV-1 nef gene products are functionally analogous, we compared the replication kinetics and infectivity of variants of SIVmac239 that either do (SIVnef+) or do not (SIV delta nef) encode intact nef gene products. SIVnef+ replicates more rapidly than nef-defective viruses in both human and rhesus peripheral blood mononuclear cells (PBMCs). As previously described for HIV-1 Nef, SIV Nef also enhances virus infectivity within each cycle of virus replication. As a strategy for evaluating the in vivo contribution of HIV-1 nef alleles and long terminal repeat regulatory sequences to the pathogenesis of immunodeficiency disease, we constructed SIV-HIV chimeras in which the nef coding and U3 regulatory regions of SIVmac239 were replaced by the corresponding regions from HIV-1/R73 (SIVR7nef+). SIVR7nef+ displays enhanced infectivity and accelerated replication kinetics in primary human and rhesus PBMC infections compared to its nef-defective counterpart. Converse chimeras, containing SIV Nef in an HIV-1 background (R7SIVnef+) also exhibit greater infectivity than matched nef-defective viruses (R7SIV delta nef). These data indicate that SIV Nef, like that of HIV-1, does enhance virus replication in primary cells in tissue culture and that HIV-1 and SIV Nef are functionally interchangeable in the context of both HIV-1 and SIV.

Animals↗
PubMed1994

Immunohistochemical changes and PCR detection of HIV provirus DNA in brains of asymptomatic HIV-positive patients.

The stage of HIV infection at which the virus enters the nervous system remains poorly understood. Examination of brains of HIV-positive non-AIDS patients often shows lymphocytic meningitis, myelin pallor and gliosis, but no immunohistochemical (IHC) evidence of the virus. In this study we have examined a number of brains from HIV-positive patients with (23) and without (8) AIDS as well as brains from 5 HIV-negative controls by morphological, morphometric, IHC and polymerase chain reaction (PCR) methods in an attempt to establish at what stage of the infection HIV can be detected in the brain and to correlate its presence with the pathological changes in the cortex. HIV-1 proviral DNA was found by PCR in the cortex of the majority of AIDS and in 2 out of 8 non-AIDS cases. Astroglial reaction was observed in the cortex of the majority of AIDS brains but not in most of the non-AIDS brains which showed, in addition, a dramatic reduction of glial fibrillary acidic protein staining around blood vessels; moreover, in this group the density of microglial cells was higher than in the AIDS group. These results show that: 1) HIV proviral DNA can be found in the brains of HIV-positive non-AIDS patients; 2) in the same group there is an increase in density of microglial cells which 3) appears to be transient, since AIDS brains without neuropathology show a lower density of these cells. They also suggest that the status of 'immune reaction' existing in AIDS may predate this period.(ABSTRACT TRUNCATED AT 250 WORDS)

AIDS-Related Opportunistic Infections↗
PubMed1993

Visualisation of arterial structure in vivo with intravascular ultrasound.

BACKGROUND: Contrast angiography provides a silhouette of the arterial lumen, but does not give information about arterial wall structure. Catheter-tip ultrasound transducers can now provide a cross-sectional image of the arterial wall. This study examined the pathological correlation of intravascular ultrasound images and the accuracy of ultrasound measurements of vascular geometry. METHODS: Intravascular ultrasound images were obtained with a mechanically rotated catheter-tip transducer and recorded on videotape. Initial validation studies were performed in fresh, post-mortem arterial specimens, which were filled with saline at physiological pressures. Ultrasound images at specific sites were compared with the pathological findings at that site and measurements of luminal diameter were compared with corresponding angiographic measurements. Subsequently, intravascular ultrasound was employed to examine the aorta, ilio-femoral and coronary arteries in patients undergoing balloon angioplasty. RESULTS: The pathological correlations showed that intravascular ultrasound can detect early initial thickening and mild atherosclerotic lesions that do not result in luminal deformation. Ultrasound images provided definition of calcified, fibrotic and lipid-filled lesions. Ultrasound measurements of luminal diameter correlated well with pathology measurements (r = 0.93), as did ultrasound measurements of plaque area (r = 0.89). The in vivo studies demonstrated that intravascular ultrasound can define atheroma lesions not evident on contrast angiography and permits detailed evaluation of the results of interventions such as balloon angioplasty. CONCLUSIONS: Intravascular ultrasound provides a unique window upon arterial structure and pathology in humans. Ultrasound images allow accurate measurements of vascular geometry and define early atheromatous lesions that are not evident with angiography.

Arteries↗
PubMed1992

The pathology of the posterior root ganglia in AIDS and its relationship to the pallor of the gracile tract.

The spinal cord and the thoracic and lumbar posterior root ganglia (PRGs) of 14 HIV-positive men and 7 age- and sex-matched controls were studied by routine histology, morphometric analysis of the number of nodules of Nageotte (nN) and the diameters of sensory ganglion cells, immunohistochemistry and in situ hybridization. In 7 patients (2 of whom had evidence of cytomegalovirus ganglionitis) there were increased numbers of nN and diffuse, mild infiltration with CD45R+ T lymphocytes; no B lymphocytes were observed. Macrophages were increased in number in all cases. Whenever more than one ganglion was examined from the same patient, the appearances were similar in all. There was no alteration in the distribution of ganglion cell diameters. Changes in the spinal cord included vacuolar myelopathy (5 cases), HIV myelitis (1 case), microglial nodules (3 cases) and pallor of the gracile tracts (GTP) in 7 cases, in 6 of whom it co-existed with increased numbers of nN. Seven cases had no abnormalities, except the increase in number of macrophages in PRGs. In spite of a correlation between sensory nerve cell loss and GTP our findings suggest that other mechanisms, such as 'dying back' may contribute to the pathogenesis of GTP. Moreover, sensory disturbances were found most commonly in association with nerve cell loss; however, loss of sensory ganglion cells was not necessarily associated with evidence of sensory impairment.

Acquired Immunodeficiency Syndrome↗
PubMed1992

Special education placements of language-disordered children in a psychiatric population.

Children evaluated by an interdisciplinary team that provided data on psychiatric disturbance, language handicaps, and psychoeducational assessment were placed in special education classes following the complete evaluation. Discriminant analysis as a special case of multiple regression analysis determined that mean reading achievement score, pre-evaluation classroom placement, and to a lesser degree, Performance IQ predicted classroom placement. Overall, correct classroom predictions approached 90 percent. These findings highlight the value of interdisciplinary evaluation to educators and mental health professionals.

Achievement↗
PubMed1992

Detection of HIV proviral DNA in cortex and white matter of AIDS brains by non-isotopic polymerase chain reaction: correlation with diffuse poliodystrophy.

OBJECTIVE: (1) To determine whether detection of HIV proviral DNA sequences in the cerebral cortex correlates with the presence of pathological changes in this region, believed to contribute to the HIV-associated cognitive/motor complex. (2) To compare the frequency with which HIV infects cortical and subcortical regions of the brain. DESIGN: In vitro studies on HIV neurotoxicity suggest that HIV may be involved in the pathogenesis of cortical damage, recently defined as diffuse poliodystrophy (DPD) in AIDS. Previous detection of HIV antigen has localized HIV more frequently to subcortical than to cortical regions. It is not known whether HIV preferentially infects subcortical tissues or if viral expression varies in these two regions. METHODS: HIV antigen and proviral DNA sequences were detected in anterior frontal lobe tissues using immunohistochemistry (IHC) and the polymerase chain reaction (PCR), respectively. DPD was assessed by staining with antibodies against astrocytes (GFAP) and microglia/macrophages (HAM 56). RESULTS: HIV proviral DNA was detected in nine out of 15 cortical samples and in 10 out of 15 white matter samples, whilst HIV p24 antigen was localized to the cortex in three out of 15 and to the white matter in seven out of 15 cases. DPD was found in 10 cases, although in five a different aetiology may have been involved. However, DPD was present in eight out of the nine cases in which HIV proviral DNA was detected in the cortex. CONCLUSIONS: Using a non-isotopic PCR method, HIV was detected in the brains of more cases than would be expected on the basis of IHC detection, and was present in the cortex as frequently as in the white matter. HIV, together with other factors, may contribute to the pathogenesis of DPD.

AIDS Dementia Complex↗
PubMed1992

Varicella-zoster virus encephalitis in acquired immunodeficiency syndrome: report of four cases.

Four patients with acquired immunodeficiency syndrome, a 27-year-old female intravenous drug abuser and three males (two drug addicts aged 27 and 33 years and a 40-year-old homosexual) presented with a rapidly progressive encephalopathy. Two had generalized varicella-zoster virus skin infection, one had had a regressive thoracic zoster rash 7 months previously and one had no history of cutaneous eruption. Neuropathological examination revealed, in each case, multifocal necrotic changes with numerous, intranuclear Cowdry type A inclusion bodies in glial cells, endothelial cells, macrophages and neurons, within and around the lesions. These inclusion bodies were stained positively for varicella-zoster virus by immunocytochemistry and contained herpes virus nucleocapsids by electron microscopy. Molecular biology using the polymerase-chain-reaction method demonstrated viral genome. In one case, zoster-induced non-inflammatory vasculopathy involved medium sized leptomeningeal vessels and was associated with circumscribed areas of cortico-subcortical infarction. In another case, varicella-zoster virus encephalitis was associated with human immunodeficiency virus encephalitis and a secondary cerebral lymphoma. Multinucleated giant cells expressing human immunodeficiency virus proteins in their cytoplasm, were found in the lymphomatous deposits and in the varicella-zoster virus necrotic lesions. In these latter lesions, Cowdry type A inclusion bodies could be seen in the nuclei of some multinucleated giant cells confirming previous observations of MGCs co-infected by HIV and CMV, and supporting the hypothesis that DNA viruses interact with HIV, thus increasing its effect.

Acquired Immunodeficiency Syndrome↗
PubMed1992

Impaired joint mobility in Guillain-Barré syndrome: a primary or a secondary phenomenon?

Three patients with Guillain-Barré syndrome had significant residual impairment of joint mobility. Pain in the limbs and axial skeleton was a prominent early feature, as were autonomic disturbances and bulbar involvement resulting in prolonged mechanical ventilation. All three patients developed marked joint stiffness and contractures despite having physiotherapy from the outset. The skeletal problems and complications became major components of disability despite improving neurological status.

Aged↗
PubMed1991

Macrophages in human sensory ganglia: an immunohistochemical and ultrastructural study.

The paper describes the immunohistochemical and ultrastructural features of normal posterior root ganglia in a group of humans aged 1 day to 80 years and compares the findings with those seen in the ganglia of normal rats of various ages, some of which underwent permanent traumatic lesions of the sciatic nerve. In humans, cells with the immunohistochemical reactions of macrophages are present in small number at birth, most of them having an endoneurial position. Subsequently, their number increases and more of them are seen around neurons, where their processes intermingle with those of satellite cells. Ultrastructural studies confirm that, in addition to interstitial cells, a small number of cells in satellite position have features of mesenchymal cells. In this respect, human sensory ganglia differ from those of rodents and this difference may explain why no nodules of Nageotte can be found either in ageing animals or after a permanent damage to the nerve has produced considerable cell loss. Other features observed in human ganglia, but absent in rats, are multiple layers of satellite cells surrounding each neuron and desmosome-like structures between satellite cell processes. Previous studies describing maturation of the satellite-nerve cell complex in animals are confirmed. In addition, the present investigation shows that, in human ganglia, satellite cells acquire a more elaborate structure than in rodents. It is also suggested that mesenchymal cells may play a role in the trophism of nerve cells and their removal after irreversible damage.

Adolescent↗
PubMed1990

The involvement of the cerebral cortex in human immunodeficiency virus encephalopathy: a morphological and immunohistochemical study.

The encephalopathy resulting from direct infection of the brain by human immunodeficiency virus (HIV), which correlates clinically with the AIDS dementia complex, has been reported as being localized to the white matter where it induces myelin loss, gliosis and perivascular infiltration by mononuclear macrophages and multinucleated giant cells. Damage to the cortical grey matter in HIV encephalopathy was investigated in nine randomly selected HIV-positive cases with or without clinical or morphological evidence of encephalopathy and in five age-matched controls, using routine histology and immunohistochemical methods [glial fibrillary acidic protein (GFAP), microglia and HIV antibodies]. Increased numbers of GFAP-expressing astrocytes and Ricinus communis agglutinin 1-120-expressing microglial cells were found in all the HIV-positive cases (including asymptomatic) and their severity could be correlated with the severity of the encephalopathy in the white matter; the increase in number of cells expressing GFAP was diffuse and the intensity of the staining higher than that of microglial cells. The subpial region was the most severely involved. It is suggested that involvement of the cortical grey matter is more common in HIV infection than previously suspected and that clinical evidence of a dementing process in AIDS is not necessarily due only to white matter lesions.

AIDS Dementia Complex↗