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S J Richards

Publications and source records attributed to S J Richards.

At least 37 records · Page 2Linked to original sources
PubMed1999

Analysis of T cells in paroxysmal nocturnal hemoglobinuria provides direct evidence that thymic T-cell production declines with age.

Peripheral blood T cells in patients with paroxysmal nocturnal hemoglobinuria (PNH) comprise a mixture of residual normal and glycosylphosphatidylinositol (GPI)-deficient PNH cells. Using multicolor flow cytometry, we demonstrated significant differences between the proportions of naive and memory cells within these populations. PNH T cells comprise mainly naive cells (CD45RA(+)CD45R0(-)), whereas normal T cells in the same patients were predominantly memory (CD45RA(-)CD45R0(+)) cells. Functional analyses showed that GPI-deficient CD45RA(+) T cells can convert to a CD45R0(+) phenotype. We present data from a PNH patient in remission for 20 years who still had significant numbers of GPI-deficient T cells; these showed a normal distribution of naive and memory components. The predominantly naive phenotype of GPI-deficient T cells seen in PNH patients with active disease likely reflects the phenotype of recent normal thymic emigrants. In patients where hematopoiesis was predominantly derived from the PNH stem cell, absolute numbers of both naive PNH CD4(+) cells and CD8(+) cells show an inverse correlation with patient age, implying this age-related decline in T-cell production is secondary to a decrease in thymic activity rather than a stem cell defect.

Adult↗
PubMed1999

Rapid expression of polymorphic ovine prion proteins and studies on their protease sensitivity.

We have used coupled and uncoupled in vitro transcription/translation to express rapidly aglycosyl ovine prion proteins from ovine genomic DNA genotyped for scrapie susceptible and nonsusceptible polymorphisms. Unlike previous in vitro studies of prion proteins, this method does not require cloning or laborious extractions. To our knowledge, this is the first report of ovine PrP expression at low (ng) levels under the control of an Escherichia coli promoter and ribosome binding site both coded for in the polymerase chain reaction primer. The rapidity of this approach could form the basis of a high throughput screening assay for PrP interactions, as proteins were expressed in a matter of hours from genomic DNA as the starting material. There was no difference observed in proteinase K sensitivity between prion translation products containing either scrapie susceptible or nonsusceptible polymorphisms.

Animals↗
PubMed1998

Lymphocyte subset analysis and glycosylphosphatidylinositol phenotype in patients with paroxysmal nocturnal hemoglobinuria.

Using multicolor flow-cytometry we have examined 19 patients with paroxysmal nocturnal hemoglobinuria (PNH) (18 with active disease and 1 spontaneous remitter) to determine absolute numbers of lymphocyte subsets and the proportion of glycosylphosphatidylinositol (GPI)-deficient clones amongst these subpopulations. Lymphocyte subsets were abnormal in all patients; the most frequent findings were low absolute numbers of natural killer (NK) cells (median, 0.08 x 10(9)/L; normal range, 0.2 to 0.4 x 10(9)/L) and low absolute numbers of B cells (median, 0.05 x 10(9)/L; normal range, 0.06 to 0.65 x 10(9)/L). GPI-deficient B, T, and NK cells were identified in 88%, 84%, and 89% of patients, respectively. The proportion of GPI-deficient cells within individual lymphoid lineages was highly variable, though in most patients the percentage of GPI-deficient NK cells was considerably higher than B or T cells. These observations can be explained when mechanisms of normal lymphopoiesis are considered. Despite these quantitative and qualitative abnormalities, no patients suffered an excessive number or severity of infections. The detection of PNH clones amongst all lymphocyte lineages may provide important information regarding the natural history of the disease and additional insights into kinetics of adult lymphopoiesis.

Adult↗
PubMed1998

Correlations of cranial morphology, ecology, and evolution in Australian suctorial tadpoles of the genera Litoria and Nyctimystes (Amphibia: Anura: Hylidae: Pelodryadinae).

Suctorial anuran larvae are highly specialized for living in fast-flowing waters, using their oral disks as adhesive organs to attach to the substrate. The cranial musculoskeletal structure of suctorial larvae of Litoria nannotis, L. rheocola, and Nyctimystes dayi (Hylidae: Pelodryadinae) were compared with congenerics with pond-type larvae (L. caerulea, L. genimaculata, L. xanthomera). Data from two other neobatrachian species with suctorial larvae (Boophis sp., Hyla armata) as well as published descriptions were taken into account. Suctorial tadpoles evolved several times independently in the Neobatrachia and share various features, irrespective of their phylogenetic position. These include the following. Cornua trabeculae are expanded anteriorly and sometimes fused. The lower jaws are robust. The greatest width of the skull is at the level of the jaw articulation. The upper jaw cartilages are partially or fully fused. The palatoquadrate is robust and connected to the skull by a wide commissura quadratocranialis anterior, processus oticus, processus basalis (in some species), and processus ascendens (vestigial or absent in some species). A processus ventralis quadrati provides an extended area of origin for the m. orbitohyoideus. The m. rectus abdominis inserts far anterior and acts on the cranium. The insertion of the epaxial musculature is shifted anteriorly to the anterior parts of the otic capsule. The mm. diaphragmatobranchialis and rectus cervicis cross at their origins. The origin of the m. levator mandibulae anterior has shifted posteriorly. The branchial basket is relatively small and the ceratohyal area is large. Multiple convergent evolution of these features suggests that they may be causally associated with the suctorial mode of larval life. Aside from these characters, however, the suctorial and pond-type neobatrachian species are remarkably similar in their jaw musculature and hyobranchial musculoskeletal composition. In some features, Ascaphus truei differs significantly from the neobatrachian suctorial species, indicating the influence of the historically distant separation of the two taxa. A novel modification of the upper jaw abduction mechanism has evolved in L. nannotis, L. rheocola, and N. dayi. It involves an adrostral cartilage as a pushing-rod element. This mechanism and unique structural similarities of the cartilago labialis superior gives support to the preliminary assumption that the nannotis species group is more closely related to the suctorial Nyctimystes dayi than it is to other Litoria species with pond-type larvae. Suctorial larvae presumably were present in the most recent common ancestor of the Litoria nannotis group and Nyctimystes dayi.

Animals↗
PubMed1998

Trisomy 12 is seen within a specific subtype of B-cell chronic lymphoproliferative disease affecting the peripheral blood/bone marrow and co-segregates with elevated expression of CD11a.

In order to delineate the specific morphological and immunophenotypic features of B-cell lymphoproliferative disorders associated with trisomy 12, 172 sequential unselected cases of CD19+CD5+ B-cell disorders, primarily affecting the peripheral blood and bone marrow, were studied. Trisomy 12 was found in 24 cases (13.9%), with all cases morphologically classified as either CLL-PL or CLL-mixed by FAB criteria. Trisomy 12 was not found in any cases of typical CLL. Trisomy 12 cases demonstrated a significant higher expression of CD11a (P<0.0001) and CD20 (P<0.0006) when compared to cases with the equivalent morphology and immunophenotype, but without the chromosomal abnormality. Trisomy 12 cases also demonstrated a higher frequency of FMC7, CD38 expression and moderate to strong surface immunoglobulin staining. However, no correlation was detected between the percentages of trisomy 12 cells and cells expressing CD11a, CD38, FMC7 or sIg mean fluorescent intensity. Cells from trisomy 12 positive cases were sorted according to their CD11a expression using fluorescent activated cell sorting. There was a significant increase in the percentage of trisomy 12 cells within the CD11a+ sorted fraction compared to the unsorted population (P < 0.05), implying that trisomy 12 is associated with increased expression of CD11a. With the highly specific morphological and immunophenotypic features demonstrated by trisomy 12 cases in this study, it is highly likely that these cases constitute a specific group of B-cell lymphoproliferative disorders.

Adult↗
PubMed1998

Granulocyte apheresis in inflammatory bowel disease: possible mechanisms of effect.

We have studied the effects of granulocyte apheresis in 18 patients with ulcerative colitis and 6 with Crohn's disease who had failed to respond to conventional therapy. Patients were treated with weekly apheresis using a granulocyte removal column (GI, Otsuka Pharmaceutical Co., Ltd., London, U.K.). We found a mean reduction in circulating granulocytes of 1.29 x 10(9) cells/L with no significant alterations in red blood cell monocyte, total lymphocyte, absolute T-helper, or T-cytotoxic lymphocyte counts. There were no significant changes in complement levels or immunoglobulin subclasses. There was a significant increase in granulocyte adhesion and a reduction in L-selectin expression. The removal of granulocytes is unlikely to explain the effect of granulocytapheresis. The markedly increased expression of alpha(m) integrin/Mac-1 and low L-selectin expression alter the capability of granulocytes to migrate to sites of inflammation and may be responsible for the improvement observed in patients treated with granulocyte apheresis.

Adult↗
PubMed1997

Five-year incidence and prediction of dementia and cognitive decline in a population sample of women aged 70-79 at baseline.

OBJECTIVE: To determine the incidence of dementia and cognitive decline and their predictors in a population sample. SETTINGS: A rural geographically defined population served by a single health centre. SAMPLE: 75% of the 70-74-year-old, 100% of the 75-79-year-old women registered with the practice. METHOD: Interview the total sample using an augmented version of CAMDEX, including informant interview, and reaction times at baseline and 5 years later. IQ was estimated at baseline only. RESULTS: 365 women were interviewed at baseline, 237 at follow-up. Incidence of dementia of all severities was 3.1% per annum (95% CI, 2.2-4.3). CAMCOG declined by 12.6 points in those who became demented, MMSE by 3 points. Observed, self-reported and informant-reported cognitive difficulty at baseline all predicted later incidence of dementia. The additional use of estimated IQ and reaction times did not increase the efficiency of either cross-sectional detection of dementia or prediction of dementia at 5 years. CONCLUSIONS: The incidence of dementia was in the range reported by other studies; slight cognitive decline was noted in the total population, increasing with age and marked in incident dementia. Of clinical importance was the finding that both the individuals and the informants of those who subsequently demented were aware of decline 5 years earlier. Simple and relatively easily administered cognitive tests were found to be the most efficient method of detection of dementia.

Aged↗
PubMed1997

Circulating plasma cells in multiple myeloma: characterization and correlation with disease stage.

The aim of this study was to develop a flow cytometric test to quantitate low levels of circulating myeloma plasma cells, and to determine the relationship of these cells with disease stage. Cells were characterized using five-parameter flow cytometric analysis with a panel of antibodies, and results were evaluated by comparison with fluorescent consensus-primer IgH-PCR. Bone marrow myeloma plasma cells, defined by high CD38 and Syndecan-1 expression, did not express CD10, 23, 30, 34 or 45RO, and demonstrated weak expression of CD37 and CD45. 65% of patients had CD19- 56+ plasma cells, 30% CD19- 56(low), and 5% CD19+ 56+, and these two antigens discriminated myeloma from normal plasma cells, which were all CD19+ 56(low). Peripheral blood myeloma plasma cells had the same composite phenotype, but expressed significantly lower levels of CD56 and Syndecan-1, and were detected in 75% (38/51) of patients at presentation, 92% (11/12) of patients in relapse, and 40% (4/10) of stem cell harvests. Circulating plasma cells were not detectable in patients in CR (n = 9) or normals (n = 10), at a sensitivity of up to 1 in 10,000 cells. There was good correlation between the flow cytometric test and IgH-PCR results: myeloma plasma cells were detectable by flow cytometry in all PCR positive samples, and samples with no detectable myeloma plasma cells were PCR negative. Absolute numbers decreased in patients responding to treatment, remained elevated in patients with refractory disease, and increased in patients undergoing relapse. We conclude that flow cytometry can provide an effective aternative to IgH-PCR that will allow quantitative assessment of low levels of residual disease.

Adult↗
PubMed1996

Immortalized neural cells from trisomy 16 mice as models for Alzheimer's disease.

The trisomy 16 mouse (Ts16) is a general accepted animal model for both Downs syndrome (DS) and Alzheimer's Disease (AD). However, the efficacy of this model is severely hampered by the fact that Ts16 is lethal after about 18-20 days of gestation. Chimeras, long-term tissue culture and neural transplantation of Ts16 material have previously been used to overcome this limitation presented by death in utero of the Ts16. In this paper we describe a new strategy to overcome this limitation, i.e. immortalization of primary cells from Ts16 mice with retrovirus-mediated gene transfer of a temperature sensitive immortalizing oncogene. By this method we have obtained a total of 21 stable cell lines from Ts16 hippocampus, Ts16 cortex, normal hippocampus, and normal cortex. So far, two of the cell lines have been karyotyped and as expected, the cell line immortalized from Ts16 embryos has retained three copies of chromosome 16. We are currently characterizing these cell lines with respect to expression of APP, T-antigen, Nestin, GFAP, NF and Map-2. Moreover, the processing and secretion of APP fragments are being investigated by immunoblotting. In summary, we have immortalized CNS cells from Ts16 mice and we expect that these cell lines will be useful as in vitro and in vivo models for studying various aspects of the pathology of Alzheimer's disease.

Alzheimer Disease↗
PubMed1996

Helminth infracommunities in Litoria genimaculata (Amphibia:Anura) from Birthday Creek, an upland rainforest stream in northern Queensland, Australia.

The helminth fauna of Litoria genimaculata, a rainforest frog from northern Queensland, was quantified from 53 adult male frogs collected at monthly intervals between April 1990 and March 1991. The helminth fauna of this species was depauperate (6 species: Mesocoelium sp., Parapolystoma bulliense, Austraplectana sp., Onchocercidae gen. sp., Cosmocerca sp. and an unidentified nematode larva). The most commonly encountered species was P. bulliense, but the intestinal infracommunity was dominated by the digenean Mesocoelium sp. Fifty-five per cent of frogs were infected with only 1 helminth species and only 1 frog had more than 2 species, resulting in low diversity values. These results support previous studies which indicate that amphibians have depauperate helminth communities.

Animals↗
PubMed1995

Influence of laparoscopic and conventional cholecystectomy upon cell-mediated immunity.

Surgery, trauma and anaesthesia induce a state of transient immunosuppression. Laparoscopic cholecystectomy has several well documented clinical advantages over traditional cholecystectomy and provokes a lower acute phase response, thought to be a result of the smaller wound size. The influence of laparoscopic cholecystectomy (21 patients) and conventional open cholecystectomy (13 patients) upon components of the cell-mediated immune system was investigated. Cell-mediated immunity was studied by in vitro assays of T lymphocyte proliferation to different mitogens, and by natural killer cell cytotoxicity using a standard 51Cr release assay. Blood samples were taken before and 24 h after the start of the operation. In the sample taken after operation there was significant depression of T lymphocyte proliferation to phytohaemagglutinin (stimulation index 149.4 versus 33.3, P < 0.002), staphylococcal enterotoxin B (85.2 versus 52.6, P = 0.01) and toxic shock syndrome toxin (48.4 versus 14.8, P = 0.08) in the group of patients who underwent open surgery, but not in the group treated by laparoscopic surgery. There was a small but statistically insignificant decrease of natural killer cell cytotoxicity in both groups of patients. These findings suggest that laparoscopic cholecystectomy causes less depression of cell-mediated immunity than open cholecystectomy.

Adult↗
PubMed1995

Immunophenotypic and DNA genotypic analysis of T-cell and NK-cell subpopulations in patients with B-cell chronic lymphocytic leukaemia (B-CLL).

Absolute numbers and distributions of peripheral blood T-cells and NK cells were immunophenotypically determined in 21 patients with B-CLL and compared with those obtained from a series of 13 elderly normal controls with an age range of 60-87 years. For absolute CD3+, CD4+ and CD8+ T-cell, and CD16+ NK subpopulation numbers, there were no consistent differences between the normal and B-CLL groups although some individual patient variation was seen. Immunophenotypic analyses did however reveal that CD3+ T-cells in almost half (10/21) of the B-CLL patients were Ia+ (defined as > 20% positive cells), compared to 0/13 of the elderly control group (p < 0.001), and that the proportions of CD4+ and CD8+ cells expressing membrane CD45RO were significantly increased compared to the control group. Subdivision of the B-CLL cases into those with low (< 20%) and high (> 20%) proportions of CD3+ T-cells co-expressing Ia further showed that CD45RO expression by CD4+ fractions was particularly prominent in the Ia+ subgroup, and that the relative increase of CD4+CD45RO+ cells was primarily a consequence of decreased absolute numbers of CD4+CD45RA+ lymphocytes. This study also examined extracted DNA from enriched CD3+ T-cell fractions (obtained by immunomagnetic bead selection in 9 of the B-CLL cases) by PCR analysis with two primers for the T-cell gamma gene locus. With the V gamma C (consensus) primer, 8/9 cases were polyclonal and the remaining case was oligoclonal. For comparison, 7/9 CD3+ fractions were oligoclonal with the V gamma 9 primer with the other two cases being polyclonal. No monoclonal CD3+ components were found. It is suggested that the observed increased Ia expression by CD3+ cells and the predominance of CD4+ cells expressing membrane CD45RO in patients with B-CLL may be of potential relevance to understanding the pathogenesis and patterns of disease progression.

Aged↗
PubMed1995

Clonal CD3+CD8+ large granular lymphocyte (LGL)/NK-associated (NKa) expansions: primary malignancies or secondary reactive phenomena?

This study reports the clinical, haematological and immunophenotypic features of a series of 25 patients with clonal expansions of large granular lymphocytes (LGL)/NK-associated (NKa) cells. These showed a male predominance (16:9) with a median age of 67 (range 38-91) years; four had a documented history of rheumatoid arthritis, a further 18 had diverse clinical disorders, and the remaining three were clinically well. Mild anaemia was found in approximately half the patients and a lymphocytosis (seen in approximately 70% of the cases) was usually modest (< 10.0 x 10(9)/l). Neutropenia was the most frequently observed feature, and this was typically persistent in nature. Serum studies revealed few consistent features although positive rheumatoid factor and increased soluble CD8 levels were noted in 67% and 87% of those cases tested. Phenotypically, all cases were CD2+CD3+CD8+ and expressed membrane TCR alpha beta chains; most (17/22) were additionally CD5+ and (19/22) CD7+. The staining intensities of CD5 and CD7 antigens were however lower than that of normal CD4+ and CD8+ blood lymphocytes. Expression of NKa antigens was variable although 16/22 cases were CD16+CD56- and 19/22 were CD57+. Clonal CD3+CD8+ LGL/NKa expansions with a CD16+CD56+ composite phenotype were not seen in this patient series. Analyses of 'activation' antigens showed a consistent lack of CD25 expression by CD3+ cells, but increased CD3/Ia co-expression was found in a high proportion (19/25) of cases. Studies of CD45R isoform expression by CD8+ LGL/NKa cell fractions revealed a consistent CD45RA+RO- profile for all cases tested.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗