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PubMed2001

The frequency of founder mutations in the BRCA1, BRCA2, and APC genes in Australian Ashkenazi Jews: implications for the generality of U.S. population data.

BACKGROUND: Several studies have shown that Ashkenazi Jews in the United States and Israel have a high prevalence of the founder mutations BRCA1 185delAG, BRCA1 5382insC, BRCA2 6174delT, and APC I1307K at frequencies of 1.0--1.1%, 0.2--0.3%, 0.6--1.4%, and 6.1--7.0%, respectively. The objective of this study was to compare the prevalence of these alleles in the Australian Jewish population with that of U.S. Jews. Australian Jews have a different history of migration, with less opportunity for changes in allele frequency due to conversion or intermarriage with non-Jewish Australians. The results obtained therefore can be used to assess whether U.S. data can be generalized to other Jewish populations. SUBJECTS AND METHODS. Subject samples were ascertained through a screening program for Tay-Sachs disease as part of a community-based screening program in New South Wales and Victoria. DNA extracted from 1200 deidentified blood samples was tested using amplification refractory mutation system polymerase chain reaction. RESULTS: The allele frequencies found were as follows: BRCA1 185delAG 1.25% (95% confidence interval [CI], 0.62--1.88%), BRCA1 5382insC 0.25% (95% CI, 0--0.53%), BRCA2 6174delT 1.08% (95% CI, 0.50--1.67%), and APC I1307K 8.67% (95% CI, 7.07--10.26%). The prevalence of breast carcinoma predisposition alleles therefore is greater than 2.5% in Australian Ashkenazim. CONCLUSIONS: There were no significant differences between the allele frequencies in Australian Ashkenazim and those identified in other studies with similar ascertainment strategies, despite the different migration patterns of Australian Jews. This suggests the broad applicability of the U.S. and Israeli data, not only to Australian Ashkenazim, but also to Ashkenazi communities throughout the world.

Adult↗
PubMed2000

Identifying migratory Salmo trutta using carbon and nitrogen stable isotope ratios.

Many Salmo trutta populations consist of non-anadromous (freshwater-resident) brown trout and anadromous (sea-run migratory) sea trout. Although adult brown trout and sea trout can usually be identified using differences in size and body colouration, it is not possible to easily identify eggs/alevins as the progeny of brown trout or sea trout. In this study we show that delta(13)C and delta(15)N, measured using a continuous flow isotope ratio mass spectrometer (CF-IRMS), can accurately identify fish eggs as the progeny of freshwater-resident (delta(13)C(egg) = -25.7 +/- 1.9 per thousand,delta(15)N(egg) = 9.2 +/- 1.8 per thousand) or migratory (delta(13)C(egg) = -19.9 +/- 1.1 per thousand, delta(15)N(egg) = 14. 3 +/- 1.5 per thousand) adult female Salmo trutta. Case studies show that stable isotope analysis is a more reliable technique for distinguishing anadromous adult fish than differentiation using morphological characteristics. For example, stable isotope analysis of brown trout from Loch Eck, Scotland, revealed that some individuals possessed delta(13)C and delta(15)N signatures indicative of marine feeding despite visual identification as freshwater-resident fish. It is most likely that these fish are misidentified sea trout although it possible that these fish may be brown trout that have adopted an estuarine feeding strategy to avoid interspecific competition for food within Loch Eck with salmon, powan and Arctic charr. Most stable isotope studies of fish ecology use terminal tissue sampling to provide sufficient biological material for isotopic analysis; however, our study suggests that adipose fin tissue could provide a comparable measure of delta(13)C and delta(15)N. Such a strategy would be invaluable when studying the trophic ecology or migration patterns of fish of high conservation value.

Adipose Tissue↗
PubMed2000

Diversity of peripheral blood mononuclear cells as revealed by a novel multiple microgel "comet assay".

Multiple microgel comet assay (MMCA) is a metho-dological adaptation of the single-cell gel electrophoresis assay in which we have introduced the use of standard agarose plug molds in an attempt to improve and expand the applications of the assay. We focused on the study of the heterogeneity of peripheral blood mononuclear cells (PBMC) at the level of the basal single-strand breakage and the DNA damage induction caused by ionizing radiation. Differences among subpopulations were also investigated at the level of chromatin organization and methylation after NotI digestion of microgel-embedded cells. In parallel experiments, the NotI-digested nucleoids were also analyzed with the use of pulsed-field gel electrophoresis (PFGE) and the DNA migration patterns were compared with the corresponding patterns from the MMCA. Significant heterogeneity in the distribution of the oxidative DNA damage, as well as intracellular variations in the NotI digestion patterns were observed in the cell population of PBMC. The combined use of both the comet assay and PFGE provides a useful model for analysis of variation in DNA damage in individual cells as well as information on size of DNA fragments.

Chromatin↗
PubMed2000

Chemokines define distinct microenvironments in the developing thymus.

During thymus development, prothymocytes home to the thymus where they migrate as maturing thymocytes from the cortex to the medulla. Chemotaxis assays show that developing T cells of newborn mice respond to certain chemokines depending on their differentiation state. In situ expression analyses indicate that the same chemokines are expressed in distinct microenvironments within the thymic stroma. Expression of chemokines is regulated temporally during embryogenesis; in the alymphoid early thymic anlage, only TECK, SDF-1 and SLC but not ELC, MDC or TARC are expressed. Fetal blood prothymocytes destined to colonize the thymus respond to the embryonic chemokines TECK and SDF-1 in chemotaxis assays with high efficacy. The in vivo significance of this finding is demonstrated by studies in the nude mouse where the thymic anlage lacks TECK and SDF-1 expression and prothymocytes home to the parathyroid anlage rather than to the thymic anlage. Developing thymocytes respond to chemokines expressed in distinct microenvironments within the thymic stroma in a way that correlates well with the previously observed migration pattern from cortex to medulla. The complexity of these chemokine-defined microenvironments increases as the thymic anlage develops to a mature thymus.

Animals↗
PubMed2001

Fabrication of quartz microchips with optical slit and development of a linear imaging UV detector for microchip electrophoresis systems.

We have developed quartz microchips for electrophoresis and a linear imaging UV detector along with the microchip. The microchips have an optical slit, which cut off the stray light in order to improve the sensitivity of UV absorption detection on the chip, at the bonding interface. They have been successfully fabricated on synthesized quartz glass substrates using the hydrofluoric acid (HF) solution bonding method. The signal level of UV absorption detection was effectively improved by applying microchips with the "on-chip" optical slit. It is also possible to improve the signal-to-noise ratio by repetitive scanning of linear photodiode array located along the separation channel, and signal averaging during elimination of the potential. Furthermore, the analysis may be performed until the separation of the target component is complete, because the real-time migration pattern of each component in the sample can be seen just as in a slab-gel electrophoresis, thus enabling a shorter analysis time.

Electrophoresis↗
PubMed2002

Theater as a mechanism for increasing farm health and safety knowledge.

BACKGROUND: The agricultural industry has one of the highest injury and fatality rates of all industries in the US. Hispanic farm workers constitute the largest percentage of farm workers on Eastern Washington farms. Literacy levels, language skills, and migration patterns need to be considered when developing farm health and safety education. Theater was chosen as a method to provide health education and farm safety training to farm workers and their families living in a three county region of Eastern Washington. METHODS: The most urgent health and safety education needs of Hispanic farm workers were identified by a series of focus groups and key informant interviews. The resulting data was used to develop four Spanish one-act plays, which were presented in each of the three counties. To test the effectiveness of theater as an educational tool each of the plays was accompanied by pre-and post-play self-report questionnaires. These were analyzed using two-tailed Chi-square (chi(2)) tests of significance and a one-tail Marginal Homogeneity statistic. RESULTS: A total of 301 persons completed pre-test post-test questionnaires; 185 were farm workers and 115 were local community members. Thirteen of seventeen questions designed to measure information gained directly from the plays showed a significant degree of positive knowledge change (P = < 0.10). Follow-up interviews conducted 2 months after play viewing suggest that participants retain some of the health and safety messages in the plays. CONCLUSIONS: The hypothesis, "appropriate farm health and safety knowledge increases as a result of attendance at a one-act Spanish play enacted by a community players' group" was validated since 13 of 17 questions showed a significant degree of positive knowledge change. Overall, participants indicated that they liked the plays, recalled the story lines easily, and would attend additional plays if given the opportunity.

Adult↗
PubMed1982

A nonparametric distance analysis of biochemical genetic data from the Aland Islands, Finland.

Biochemical genetic data from 3272 individuals from the Aland Islands, Finland, are analyzed using a recently developed set of nonparametric genetic distance measures. These measures are more robust than the traditional methods used in previous studies of this population. While there was general agreement in the results of the traditional and nonparametric approaches, some important differences were seen. In these cases, the nonparametric methods gave result more congruent with population history, geographic pripinquity, and migration patterns. Heterozygosity measures were also calculated for the Aland Islands population and compared to values obtained for Jewish populations. The total heterozygosity values were very similar.

Finland↗
PubMed1987

Genetic structure of the Utah Mormons: isonymy analysis.

Isonymy analysis is reported for a sample of 188,895 marriages extracted from the Utah Genealogical Database. Inbreeding rates estimated by isonymy are low, ranging from 0.005 for the earliest marriage cohort (1800-1809) to 0.0008 in the most recent cohort (1950-1959). The inbreeding values decrease considerably through time, but they are consistently higher than inbreeding values estimated from pedigrees. Several explanations are offered for this, including polyphyletism of surnames and the presence of Scandinavian patronyms in this population. Random isonymy between subdivisions is also compared with random kinship estimated from migration matrices. In terms of within-subdivision kinship, the two approaches yield similar results. However, the results are quite dissimilar for between-subdivision kinship. This reflects the recent and nonrandom settlement of Utah by different ethnic groups with different surname distributions. In later time periods, the correlations between the two types of kinship estimates increase, showing that migration patterns (which are strongly determined by geographic distance) exert an increasing influence on the distribution of surnames. Logistic regression is performed on a subset of marriages (n = 88,202), using isonymous vs. nonisonymous marriage as the dependent variable. The independent variables are year of marriage, geographic distance between husband's and wife's birthplaces, endogamous vs. exogamous marriage, and population sizes of husband's and wife's birthplaces. Year of marriage and geographic distance are shown to be significant independent predictors of isonymous marriage.

Christianity↗
PubMed1988

Inbreeding and genetic disease in Sottunga, Finland.

The contribution of inbreeding to the prevalence of recessive genetic diseases in the Aland Island parish of Sottunga is investigated. Genealogical data for 3,030 individuals spanning up to 15 generations were used to estimate inbreeding. This small island community shows a low average inbreeding value of .0031 for the period 1725-1975. A cohort analysis shows that inbreeding increased from 1750 to 1900, when maximum inbreeding for those born in Sottunga reached .0057. A sharp decline in inbreeding occurred thereafter. Individuals with island-born parents made the largest contributions to inbreeding in all time periods compared to those with one or two migrant parents. These trends are consistent with changing migration patterns and isolate breakdown in Aland since 1900. An analysis of pedigree development demonstrates that remote consanguinity contributed more to inbreeding through time than close consanguinity. Both the number of common ancestors and the number of paths of relationship between spouses increased dramatically through time, the latter at a much faster rate. The contribution to average inbreeding per path, however, diminished rapidly through time. This analysis indicates that inbreeding does not account for the high incidence of autosomal recessive disorders, such as tapetoretinal disease, found in the parish.

Consanguinity↗
PubMed1988

Statistical analysis of the migration component of genetic drift.

Statistical methods are introduced for analysis of the migration component of genetic drift, i.e., of the stochastic changes that affect allele frequencies during migration between local groups. Attention focuses on alpha M, a parameter that measures the extent to which this component of drift departs from the ideal of independent random sampling, and which can be interpreted as a measure of the extent to which migration is kin-structured. It is shown that alpha M can be estimated from genetic data, even in the absence of information about the genealogical relationships of migrants, and Monte-Carlo simulations are used to approximate the sampling distribution of the estimator under the null hypothesis of independent random sampling. Application of these methods to data from the Aland Islands, Finland, shows that the migration pattern there is consistent with the hypothesis of independent random sampling.

Finland↗
PubMed1996

High frequencies of identical T cell clonotypes in synovial tissues of rheumatoid arthritis patients suggest the occurrence of common antigen-driven immune responses.

OBJECTIVE: To investigate T cell antigen receptor (TCR) clonotypes in rheumatoid arthritis (RA) lesions. METHODS: Reverse transcriptase-polymerase chain reaction with TCR V beta family-specific primers and subsequent single-strand conformation polymorphism (SSCP) analysis were performed. Direct nucleotide sequencing was also conducted. RESULTS: A distinct clonal expansion of T cells was observed in the synovium. Furthermore, identical bands in samples of different areas of the same lesion were obtained by SSCP analysis. Nucleotide sequencing revealed that T cell clonotypes of identical mobility on SSCP analysis had the same nucleotide sequence and thus were identical clones. In 6 RA patients, 60-100% of the expanded T cell clonotypes had identical migration patterns in 2 different samples, indicating that this percentage represents commonly existing T cell clonotypes in the affected joint. Furthermore, the J beta 2.1 gene segment was used predominantly by the TCR V beta clonotypes that commonly expanded in the different portions of the same joint. CONCLUSION: These results suggest that the immune response in RA is not random, but rather is driven by common stimuli.

Adult↗
PubMed2003

Neural crest and cardiovascular development: a 20-year perspective.

BACKGROUND: Twenty years ago this year was the first publication describing a region of neural crest cells necessary for normal cardiovascular development. Ablation of this region in chick resulted in persistent truncus arteriosus, mispatterning of the great vessels, outflow malalignments, and hypoplasia or aplasia of the pharyngeal glands. METHODS: We begin with a historical perspective and then review the progress that has been made in the ensuing 20 years in determining the direct and indirect contributions of the neural crest cells, now termed cardiac neural crest cells, in cardiovascular and pharyngeal arch development. Many of the molecular pathways that are now known to influence the specification, migration, patterning and final targeting of the cardiac neural crest cells are also reviewed. RESULTS: Although much knowledge has been gained by using many genetic manipulations to understand the cardiac neural crest cells' role in cardiovascular development, most models fail to explain the phenotypes seen in syndromic and non-syndromic human congenital heart defects, such as the DiGeorge syndrome. CONCLUSIONS: We propose that the cardiac neural crest exists as part of a larger cardiocraniofacial morphogenetic field and describe several human syndromes that result from abnormal development of this field.

Abnormalities, Multiple↗
PubMed2002

The corticostriatal junction: a crucial region for forebrain development and evolution.

Most parts of the brain are conserved across reptiles and birds (sauropsids) and mammals. Two major qualitative differences occur in the upper part, or pallium, of the telencephalon, the most rostral part of the brain. Mammals have a six-layered neocortex and also exhibit a different morphological organization in the lateral half, or sector, of their pallium than do sauropsids. These differences of lateral pallial construction may derive from small but crucial differences in migration patterns of neuronal precursors generated at or above the corner of the lateral ventricle, the corticostriatal junction (CS). Sauropsids have a large structure, the dorsal ventricular ridge, that is proliferated from this region, and its anterior part (ADVR) receives ascending projections from the dorsal thalamus. Mammals have multiple structures in this same region-the lateral part of neocortex, amygdala, and claustrum-endopiriform formation. We propose here that, as the degree of development of structures that form the deeper tier of the pallium varies across the stages of embryology and across phylogeny, mutations may have occurred during evolution at the origin of mammals that had profound consequences for the fate of neural populations generated in the region of the CS and its neighboring pallial germinal zone.

Animals↗
PubMed1995

Organization of permanent and transient neuropeptide Y-immunoreactive neuron groups and fiber systems in the developing hamster diencephalon.

The development of neuropeptide Y-immunoreactive (NPY-IR) cell and fiber systems in the hamster diencephalon was studied. Eight perinatal groups of NPY-IR neurons develop into 12 distinct sets in nuclei of the adult diencephalon and mesencephalon. NPY-IR neurons of the thalamic precommissural nucleus, nucleus of the optic tract, and olivary pretectal nucleus are derived from the superior group. Those in the adult magnocellular nucleus of the posterior commissure and deep mesencephalic nucleus are from the dorsal group. An arcuate group contributes neurons to the arcuate nucleus and median eminence and a mammillary group transiently exists in the mammillary region. A medial group gives rise to two sets of neurons, one that migrates to the intergeniculate leaflet and another that develops in the medial nucleus reuniens. A very large ventral group provides NPY-IR neurons to the adult medial zona incerta and caudal reticular thalamus. Groups of NPY-IR neurons also appear in the bed nucleus of the stria terminalis and centromedian thalamic nucleus. Superior group neurons may undergo apoptosis. In several groups, neurons become fewer during development, and NPY-IR may disappear. NPY-IR neurons of several groups initially migrate away from the neuroepithelial zone with later emergence of a distinct, persistent set of NPY-IR neurons in the same neuroepithelial region. The data show that neuropeptide content can be used to identify particular sets of neurons early in development, thereby allowing migration patterns to be followed and principles of brain development to be elucidated.

Animals↗
PubMed1989

Activation of human T cells by toxic shock syndrome toxin-1: the toxin-binding structures expressed on human lymphoid cells acting as accessory cells are HLA class II molecules.

Toxic shock syndrome toxin-1 (TSST-1)-binding assay using 125I-labeled TSST-1 showed the presence of specific TSST-1 binding in a B cell fraction of human peripheral blood mononuclear cells and L cells transfected with DR2 genes or DR4 genes but not in a T cell fraction and control L cells. Fixation with paraformaldehyde, an inhibitor of antigen processing, did not remove TSST-1-binding activity of the transfectants. Binding of 125I-labeled TSST-1 to the transfectants was reduced by an anti-DR monoclonal antibody. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed the presence of a single band with TSST-1-binding activity and the same migration pattern as DR heterodimers. TSST-1-induced T cell responses, proliferation and interleukin 2 (IL2) production were observed in the presence of the transfectants but not in the presence of control L cells, while concanavalin A-induced IL2 production was observed in the presence of either the transfectants or control L cells. Presence of an anti-DR monoclonal antibody inhibited the TSST-1-induced responses. Paraformaldehyde-fixed Daudi cells were effective in supporting TSST-1-induced IL2 production by T cells. These results indicate that HLA class II molecules directly bind intact TSST-1 and perform an essential role as the TSST-1-binding structures on accessory cells in T cell activation by the toxin.

Antigen-Presenting Cells↗
PubMed1993

Beta 2-adrenergic stimulation causes detachment of natural killer cells from cultured endothelium.

Physical exercise, mental stress, or infusion of beta-adrenergic agonists result in an increase in the number of natural killer (NK) cells in the peripheral circulation. In view of the specific migration pattern of NK cells in vivo, it has been suggested that these cells may be released from the marginating pool in blood vessels. In the present report, the in vitro effect of catecholamines on the adhesion of NK cells to unstimulated human endothelial cells (EC) was characterized. Peripheral blood mononuclear cells were allowed to adhere to monolayers of EC, after which the adherent lymphocyte fraction was analyzed phenotypically by flow cytometry. NK cells were found to adhere preferentially to EC, a process that was reversed by the addition of various adrenergic agonists. Catecholamines selectively affected adhesion of NK cells and had no effect on T cell adhesion to EC, as was determined by the use of purified cell populations. Detachment of NK cells from EC could be achieved by short incubations (5 min) with epinephrine (EPI) and was concentration-dependent, with an ED50 of 2 x 10(-10)M. Using a panel of alpha- and beta-adrenergic agonists and antagonists, we show that the detachment of NK cells is mediated via beta 2-adrenergic receptors. In line with the lower affinity for beta 2-adrenergic receptors, norepinephrine was less effective than EPI in inducing detachment of NK cells from EC. Direct activation of adenylate-cyclase with forskolin gave similar results as observed with EPI, indicating that signaling through cAMP is necessary to induce detachment of NK cells from EC. The results of the present study lend support to the hypothesis that catecholamines, via beta 2-adrenergic receptors, can induce recruitment of NK cells from the marginating pool to the circulating pool, by changing the adhesive interactions between NK cells and EC.

Adenylyl Cyclases↗
PubMed1994

Evidence for cell surface association of CD2 and LFA-1 (CD11a/CD18) on T lymphocytes.

Previous studies have reported an association of the cell surface adhesion molecule CD2 with the T cell receptor and with CD45 on mouse and human T lymphocytes. In this study the association of CD2 with cell surface molecules was investigated using cell surface biotinylation of T lymphocytes, coupled with immunoprecipitation using two CD2-specific monoclonal antibodies (mAb) (RM2-5 and 12-15) and analysis by SDS-PAGE. Although both CD2 mAb immunoprecipitated CD2 from lysates of murine lymphocytes, it was found that mAb 12-15, but not RM2-5, co-precipitated two other molecules of 95 and 180 kDa. Subsequent studies revealed that the 95- and 180-kDa molecules were associated with a subspecies of CD2 (approximately 5%) on thymocytes, the antigen-specific T cell line D10, and splenic T cells but not B cells. Two lines of evidence were obtained consistent with the 95- and 180-kDa molecules being the beta and alpha chains of LFA-1. Firstly, an analysis of 12-15 mAb immunoprecipitates on 4-12% gels under reducing and nonreducing conditions shows that the 95- and 180-kDa molecules have a molecular weight and migration pattern identical to LFA-1. Secondly, depletion of LFA-1 from lysates with LFA-1 mAb abolished the ability of CD2 mAb 12-15 to co-precipitate the 95- and 180-kDa molecules, thereby identifying these as the beta and alpha chains of mouse LFA-1, respectively. These results provide evidence for the first time for an association of LFA-1 and CD2 on mouse T lymphocytes, and suggest that the association occurs with an immunologically distinct subspecies of CD2 molecules.

Animals↗
PubMed2004

Homing of immature thymocytes to the subcapsular microenvironment within the thymus is not an absolute requirement for T cell development.

T cell development is thought to occur in distinct microenvironments within the thymus. Namely, the subcapsular zone, the cortex and the medulla have been described to support expansion of the immature thymocyte pool, positive selection of useful specificities and elimination of potentially self-reactive specificities, respectively. Consistent with this model, thymocytes show a highly ordered migration pattern and move into these niches in the expected sequence. Here we show that the chemokine receptor CCR9 plays a nonredundant role in the homing of immature thymocytes to the subcapsular zone. In CCR9-deficient mice, T cells in early stages of development do not accumulate in their physiological microenvironment underneath the thymic capsule and are instead homogeneously distributed across the thymic cortex. Remarkably, this abnormality does not result in a detectable defect in T cell development in CCR9-deficient mice, suggesting that the transit of immature thymocytes through the subcapsular microenvironment is not an absolute requirement for proper T cell development.

Animals↗