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Characterization of monoclonal antihuman small bowel and colon specific mucin antibodies.

Three human small bowel and colon mucosal specific monoclonal antibodies with distinct morphologic and electrophoretic characteristics were generated by fusion of immunized Balb/c spleen cells and murine plasmacytoma cells. Morphologic specificity by indirect immunofluorescence (IIF) revealed three antibody binding patterns corresponding to villus surface (TP-NG-43), goblet cell apical granules (TP-NG-2), and a combined surface/goblet cell apical granule antibody (TP-NG-20). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) produced three distinct electrophoretic migration patterns. These antibodies reacted with very high molecular weight determinants: TP-NG-2, one band greater than 400 kD; TP-NG-20, two bands corresponding to 370-400 kD; and TP-NG-43, two bands in the 350-400-kD range with smaller bands in the 50-94-kD range. Cross-reactivity with various other human organ systems was evaluated by indirect immunofluorescence and SDS-PAGE electrophoresis with Western blotting. By IIF, all three monoclonal antibodies reacted very strongly with components of gastric mucosa. Weak cross-reactivity was seen with colon, rectum and mucin-producing adenocarcinoma of the colon. No cross-reactivity was observed by IIF with other mucin-containing and non-mucin-containing tissues. However, cross-reactivity with gastric mucin was not detected by enzyme-linked immunosorbent assay (ELISA) and Western blotting. Antibody reactivity with mucin was confirmed by purifying various regional gastrointestinal mucins and by subsequent testing by ELISA. Monoclonal antibody affinity columns were prepared and evaluated. The utility of these methods will allow for further definition of important goblet cell mucin glycoprotein characteristics and isolation of mucin subpopulations.

Antibodies, Monoclonal

Patterns of immunoreactive pancreatic polypeptide in human plasma.

Pancreatic polypeptide (PP) is synthesized as an amino-terminal moiety of a precursor peptide and is released into plasma during stimulation as an amidated hormone (PP1-36). The purpose of this investigation was to ascertain the immunoreactive forms of PP in human plasma using HPLC chromatographic technique. Plasma was obtained from five normal volunteers under various postprandial intervals and from the Blood Bank. PP in each plasma sample was processed for HPLC analysis by immunoprecipitation and/or immunoaffinity extractions. Migration patterns of PP-forms were identified under isocratic elution. This study shows that human plasma contains four distinct immunoreactive (IR) forms of PP during stimulation by a protein-rich meal. These forms are PP1-36 (peak 4), PP3-36 (peak 3) and unidentified material migrating as peak 2 and peak 1. The corresponding migration constants were Kav 0.828 +/- 0.04, Kav 0.790 +/- 0.003, Kav 0.570 +/- 0.009 and Kav 0.409 +/- 0.007, respectively. The predominant fasting from of IR PP chromatographed as peak 1, while peaks 2 and 4 were reduced in amplitude. The 1 h and 3 h postprandial chromatograms of HPLC profiles of plasma PP were similar in shape but lower in relative magnitude and amplitude. The authenticity of peak 4 as the migration of native PP1-36 was confirmed using purified IR native PP1-36 extracted from human pancreas. Partial amino acid sequence analysis of PP peak 3 revealed deletions of two N-terminal amino acid residues. The chemical identities of peaks 1 and 2 are unknown but appear to differ from PP in peaks 3 and 4 by virtue of their migration profiles. It is concluded that there are at least four distinct IR forms of PP in human plasma. Native PP1-36 accounts for less than 1% of total PP after an overnight fast and is about 1/3 of total postprandial IR plasma PP. Discernment of the nature and etiology of forms of PP in plasma may provide a new understanding of the role of PP in mammalian physiology.

Adult

DNA relatedness, karyotyping and gene probing of Candida tropicalis, Candida albicans and its synonyms Candida stellatoidea and Candida claussenii.

Isolates of Candida albicans with varied phenotypes, including sucrose-negative variants (C. stellatoidea, serotypes A and B) and avirulent germ tube-negative forms (C. claussenii) showed significant (greater than 90%) DNA relatedness to classical C. albicans, but insignificant relatedness to C. tropicalis and sucrose-negative C. tropicalis. A transverse alternating-field gel electrophoresis procedure (TAFE) showed discrete karyotype patterns among the phenotypic variants of C. albicans including the sucrose-negative C. stellatoidea. The number of chromosome-sized DNA bands for C. tropicalis (7 bands) were within the range of bands observed for C. albicans (5 to 10 bands). The general DNA-migration pattern for C. albicans appeared distinct from that of C. tropicalis. An aspartyl proteinase (PrA) gene probe from C. albicans hybridized with chromosomal DNA from C. albicans, C. claussenii and C. stellatoidea but not with that from C. tropicalis.

Aspartic Acid Endopeptidases

Skin penetration of infective hookworm larvae. III. Comparative studies on the path of migration of the hookworms Ancylostoma braziliense, Ancylostoma ceylanicum, and Ancylostoma caninum.

The lateral skin areas of nine hookworm-free puppies were exposed to infective larvae of Ancylostoma braziliense, Ancylostoma ceylanicum and Ancylostoma caninum. Serial sections, stained with Harris' haematoxylin and eosin, showed that many similarities exist in the migration patterns of the hookworm species used. However, striking differences were observed between Ancylostoma braziliense on the one hand and Ancylostoma ceylanicum and Ancylostoma caninum on the other hand with regard to the speed with which the penetration occurred. Larvae of Ancylostoma braziliense may travel directly from the epidermis into the dermis--apart from migration via hair follicle systems--a path of migration we did not observe in infections with the other species used. Larvae of Ancylostoma braziliense seem to pass the skin more rapidly than those of Ancylostoma ceylanicum and Ancylostoma caninum. Many larvae of Ancylostoma ceylanicum managed to penetrate the skin 2--6 h after the onset of the exposure.

Ancylostoma

Transcultural nursing and health care issues in urban and rural contexts.

The distinction between urban and rural health care has been based largely on population density, with little attention given to contextual meaning of patterns related to the lifeways of the people. When considering sociocultural factors, the differentiation becomes more complex. In this paper some of the transcultural nursing issues and challenges are discussed in relation to cultural context and health care trends in urban and rural settings. Four urban-rural health-related issues with transcultural dimensions are examined. Migration patterns, diversity among rural communities, utilization of services, and change related to intervention strategies are identified as transcultural nursing knowledge and practice in relation to urban and rural contexts.

Health Services Needs and Demand

A bifunctional genetic regulatory element of the rat dopamine beta-hydroxylase gene influences cell type specificity and second messenger-mediated transcription.

Dopamine beta-hydroxylase, the enzyme which converts dopamine to norepinephrine, is expressed in a cell type-restricted pattern in neuroendocrine tissue. A segment of the rat gene containing 395 bases of 5'-flanking sequence regulates expression of a reporter gene in a cell type-selective pattern in mammalian cell cultures. Using deletion mutants of the 5'-flanking sequence, we have identified a 30-base genetic regulatory element, designated DB1, which enhances transcription from a heterologous promoter 5-20-fold in neuroendocrine cell lines. DB1-specific DNA-protein complexes are found in nuclear extracts from all cell lines examined, but the migration pattern differs between cell lines. The 5'-flanking region of the dopamine beta-hydroxylase gene is also responsive to cyclic AMP and phorbol ester treatment of SHSY-5Y neuroblastoma cells. The simultaneous presence of both effectors results in synergistic increases in DBH1 mRNA and reporter gene activity. The second messenger regulatory element was localized to the region containing the DB1 element, and reporter plasmids containing multiple copies of the DB1 element are responsive to treatment with inducers. The results of this study identify a cis-acting regulatory element which influences both cell type selectivity and second messenger responsiveness of the rat dopamine beta-hydroxylase gene.

Animals

Expression of cytokine genes, cytokine receptor genes, and transcription factors in cultured Hodgkin and Reed-Sternberg cells.

In the present study, we show by Northern blot analysis and enzyme linked immunosorbent assay that the Hodgkin's disease (HD)-derived cell lines HDLM-2 and KM-H2 express a variety of cytokine genes either constitutively or upon induction with phorbol ester 12-O-tetradecanoylphorbol-13-acetate. Cytokine genes expressed by HD-derived lines include granulocyte-macrophage colony-stimulating factor (CSF), macrophage-CSF, interleukin (IL)-1-alpha, IL-3, IL-5, IL-6, IL-8, leukemia inhibitory factor, tumor necrosis factor-alpha, tumor necrosis factor-beta, and transforming growth factor-beta, while transcripts and the corresponding proteins for granulocyte-CSF, IL-1-beta, IL-2, IL-4, IL-7, IL-10, and the JE/macrophage chemoattractant and activating factor gene were not detectable in cytoplasmic RNA and culture supernatants obtained from both lines. In addition, IL-2 receptor (R) p55 and macrophage-CSF R (c-fms) genes were expressed by both lines. HDLM-2, but not KM-H2 cells, exhibited the IL-6 R p80 and the IL-2 R p75 chain. Analysis of nuclear proteins that bind to oligonucleotides containing the consensus sequences of the transcription factors activation protein 1, nuclear factor (NF) kappa B, and NFAT 1 revealed a pattern for HD lines resembling that of activated T-cells: HDLM-2 and KM-H2 cells constitutively expressed NF binding to the NF of activated T-cells (type 1), previously described to be T-cell specific. In addition, NF kappa B-binding proteins obtained from both lines showed, in electrophoretic mobility shift assays, the same migration pattern as T-cell-derived proteins but differed from monocyte- and B-cell-derived proteins. UV cross-linking experiments confirmed that NF kappa B-binding proteins of M(r) 85,000, 75,000, and 50,000/55,000 were detectable in nuclear extracts obtained from T-cells and both HD lines, while monocytes and B-cells displayed the M(r) 50,000/55,000 and 75,000 NF kappa B complex only. Both HD lines also constitutively expressed transcripts for c-fos and c-jun, which are involved in heterodimeric formation of the transcription factor activation protein 1, as well as for the NF kappa B/KBF1 gene.

Antigens, CD

Epithelial migration on the tympanic membrane. An experimental study.

The existence of an epidermal migration pattern was studied in mice using a whole-mount autoradiographical technique. The mitotic activity appeared to be confined to an area close to the annulus tympanicus. A very slow migratory activity of the labelled cells, was observed, situated in the basal cell layer. Perforation of the tympanic membrane showed no effect on the position of the labelled cells. This slow migration described by several authors. The latter migration was suggested to represent a mechanism for cleaning the tympanic membrane.

Animals

Genomic reconstruction of the Pakistani Roma reveals dual South Asian ancestry, medieval bottlenecks, and the early dispersal routes of the Romani people.

The Roma people represent one of the largest and most historically enigmatic diasporas in Eurasia, illuminating human migration patterns and cultural resilience across continents. Despite extensive research on European Roma as the diaspora endpoint, the genetic legacy of their putative South Asian source populations remains critically underexplored, leaving fundamental gaps in understanding the pre-diaspora demographic structure and early dispersal dynamics. This study uniquely positions Pakistani Roma as a potential ancestral reservoir, offering a rare window into the pre-migration phase distinct from derived European Roma populations shaped by centuries of post-dispersal admixture. We analyze 82 Pakistani Roma from Punjab using high-resolution genome-wide SNP data and comprehensive mitochondrial haplogroup profiling to reconstruct their genetic origins, population structure, and historical trajectory. Analyses reveal a dual ancestry profile comprising 50-82% Indus Valley related, 20-30% Onge related, and up to 26% Steppe derived components, with three distinct subgroups exhibiting varying affinities along a South Asian to Central Western Eurasian continuum reflecting jati-like endogamy. A severe demographic bottleneck ~800 years ago coincides with medieval socio-political upheavals, while major Eurasian admixture is dated to ~660 years ago. Mitochondrial haplogroups H (45.12%) and M (26.83%) underscore dual maternal influences from West and South Eurasia. Pakistani Roma retain substantially higher South Asian ancestry than their European counterparts, establishing them as a genetically distinct population preserving the ancestral pre-diaspora state. These findings redefine the Romani origin narrative and underscore the critical value of understudied South Asian minorities in reconstructing complex human migration pathways and diaspora formation mechanisms.

Humans

Homing and progression patterns of childhood acute lymphoblastic leukemias in severe combined immunodeficiency mice.

The aim of this study was to analyze the homing and progression patterns of childhood acute lymphoblastic leukemias (ALL) in mice with severe combined immunodeficiency (SCID). Upon intraperitoneal (IP) transfer, cells from relapse samples of three children with T-lineage ALL spread hematogenously and infiltrated the non-lymphoid and/or lymphoid organs with a pattern reminiscent of the human clinical disease. These mice either died or were killed in extremis at a mean of 9 weeks. Moreover, cell lines established in vitro from two of these samples manifested identical homing and progression in the SCID mouse as compared with the original patients' cells. Thus, long-term culture of the primary leukemic T cells did not alter their invasive potential and migration pattern. When engrafted IP, three cell lines established from pre-B-ALL cases displayed primarily a lymphatic spread with induction of local tumor masses and kidney/liver nodules. Mice were killed at 11 to 13 weeks, but had not developed imminently fatal leukemia. However, when transferred intravenously, one pre-B ALL cell line was able to spread hematogenously and to infiltrate both lymphoid and non-lymphoid tissues. Overall, these data demonstrate that the SCID mouse provides an efficient and reproducible model to study the pathogenesis of childhood ALL, and may be a suitable system for evaluating therapy.

Animals

Serial scintigraphic imaging for visualization of passive transport processes in the human fallopian tube.

Spontaneous migration of a radionuclide tracer from the vagina to the peritoneum may be visualized by scintigraphic imaging (hysterosalpingoscintigraphy, HSS). A prospective study was designed to evaluate diagnostic criteria for normal tubal passage of a control group (n = 7) and to establish the predictive value of the HSS technique in defining functional deficiency in anatomically patent tubes. In 56 patients with tubal and unexplained infertility, a comparison between the results of the tracer migration study and of contrast hysterosalpingography and laparoscopy was made. The overall correlation was 65%. Clearly discrepant results (i.e. an abnormal migration pattern in anatomically patent tubes) were recorded in 18% and were positively, yet not significantly associated with tubal adhesive disease and with a history of tubal microsurgery. Interpretation of scans was equivocal in another 18% of patients due to undetectable ascension of the tracer to the uterus. It is suggested that the radionuclide is moved forward by the same passive transport processes which are concerned with support of the migration of spermatozoa to the ovum, and that failure of tubal migration of the tracer may render patients eligible for in-vitro fertilization/embryo transfer treatment.

Adult

Evaluation of the possible role of B cell receptors in the tendency of B cells to migrate into follicles in mice and chickens.

Chicken spleen and bursa cells were examined for the percentage of Fc receptor-bearing cells. Rosette formation was done with chicken 7S antibody-sensitized sheep erythrocytes and was inhibited by heat-aggregated chicken Ig. In the spleen, the percentage was found to increase with age to approximately 26% at 7 to 12 weeks. In contrast, only 3 to 5% of bursa cellss at this age demonstrated Fc receptors. Sleens from bursectomized chickens had 7--10% Fc receptor-bearing cells. In an attempt to determine a possible role of the C3 receptor on migration patterns, the effect of cobra venom factor (CVF) on the localization of transferred lymphoid cells was examined. Pretreatment of recipients with enough CVF to lower mean C3 levels to 11% of controls failed to affect follicular B cell localization in mice at either 24 or 48 h after transfer. Localization of thymus or bursa cells in chickens was similarly unaffected by CVF pretreatment. The possible roles of Fc and C3 receptors on migration of B lymphocytes into follicles and germinal centers were discussed.

Animals

Development of of macrophage migration inhibition in rabbits infected with virulent Treponema pallidum.

Peritoneal exudate cells from rabbits infected with Treponema pallidum Nichols were used as indicators of macrophage migration inhibitory factor activity. Between 3 and 15 weeks after infection, the migration of peritoneal exudate cells was inhibited in the presence of 3 to 25 microgram of T. phagedenis biovar Reiter protein per ml. Before this period, the migration patterns of peritoneal exudate cells from infected animals were uninhibited and similar to those from noninfected control rabbits. These observations were correlated with the development of active cell-mediated immunity during experimental T. pallidum infection.

Animals

C4 reference typing report.

Human C4 is most polymorphic at the protein level, distinction between allotypes of the C4A and C4B proteins resting on electrophoretic migration patterns and difference in hemolytic activity. The aim of the C4 reference typing has been the definition of reference variants, the assignment of rare variants, and the investigation of duplicated, deleted, or non-expressed and hybrid genes. Samples from 136 individuals, predominantly with known segregation, from 16 laboratories were investigated by standard electrophoretic techniques, for their relative hemolytic activity, reactivity with monoclonal antibodies and Rg/Ch reagents, alpha-, and beta-chain types, relative electrophoretic migration distance, as well as the C4/21-OH-TaqI RFLPs. The results were evaluated in three groups; they consisted in the definition of the eight most common C4 alleles, and the ten Rg/Ch standard phenotypes in group I. In group II twelve C4A and fourteen C4B duplications among 96 complotypes, as well as eighteen deleted/non-expressed C4A and twenty-two C4B alleles, and hybrid alleles were seen by correlation of lytic activity, electrophoretic mobility, and monoclonal and/or Rg/Ch reactivity. Group III consisted of the newly defined allotypes A 8, A 7, A 58, A 55, A 45, B 45, B 35, and B 22, furthermore of alleles subdividing the A 1/A 91, and the B 13/B 12/B 11 regions. The reference typing has allowed reclassification of the majority of described C4 allotypes and resulted in a revision of the C4 nomenclature.

Blood Grouping and Crossmatching

Quantitative studies on Heterakis gallinarum infections in the common fowl, Gallus gallus L.

The fertility, mortality, and migration patterns of Hetarakis gallinarum were studied in chickens with concomitant Parahistomonas wenrichi infections. H. gallinarum females were found to produce approximately 936 ova per day, when 50 days of age, and a total of 34,000 to 86,000 ova in a lifetime. There was no evidence of differential mortality between the sexes, nor of a preference for either the left or the right caecal organ of chickens. Both male and female worms are capable of migrating between caeca, and are expecially prone to do so when in the absence of individuals of the opposite sex.

Animals

Synthesis of the acid-soluble proteins in early cleaving embryos of the sea urchin. Cyclic synthesis of histones.

Synthesis of the acid-soluble proteins in the early cleavage stage of the sea urichin, Hemicentrotus pulcherrimus, was investigated. As detected by the incorporation of lysine, the acid-soluble proteins were synthesized periodically even before the first cleavage, differing from the pattern of incorporation of tryptophan into the fraction. Cyclic synthesis occurred almost in parallel with DNA synthesis. However, the phase and periodicity of cyclic synthesis of the acid-soluble protein fraction were quite different from those found in the hot TCA-insoluble (acid-insoluble) protein fraction. The acid-soluble proteins were adsorbed on cation exchange resin, Amberlite CG-50, and gave an elution profile similar to that found for calf thymus histones. The migration pattern of these proteins on acrylamide gel also resembled that of histones.

Animals

Maturing thymocytes in accelerated rejection of cardiac allografts in presensitized rats.

LBNF1 cardiac allografts are rejected within 36 hr in LEW rats sensitized with BN skin grafts 7 days earlier (acute rejection in unmodified hosts = 8 days). We have studied and compared the function and migration patterns of thymocytes one day after engraftment in sensitized recipients, unmodified hosts, and normal naive rats. Thymocytes from animals experiencing accelerated rejection were more mature and functionally active, as shown by a significant elevation in percentage of OX-44+ (CD37+) cells, increased alloreactivity to BN and WF antigens, and proliferative responses to Con A and exogenous IL-2. However, the cells could neither lyse BN targets in vitro nor trigger rejection of otherwise indefinitely functioning test cardiac allografts in immunologically unresponsive T cell-deficient (B) rats after adoptive transfer. The traffic of 111In-labeled thymocytes was then evaluated. The migration index increased significantly during accelerated graft rejection, with thymocytes preferentially circulating in the blood, penetrating peripheral lymph nodes--and, interestingly, migrating back to the thymus. Thus, immunoresponsive and functionally active thymocytes, which lack the ability to recognize primed specific antigen, appear during accelerated rejection of cardiac allografts in sensitized rats. These cells migrate to the periphery, and then return in large numbers to their site of origin, the thymus. Hence, this study describes a novel behavior of thymocytes in the state of host alloreactivity that is distinct from the physiological one in otherwise normal thymus.

Animals

Kinetics of liver trapping of infective larvae in murine toxocariasis.

Mice sensitized by prior infection with Toxocara canis eggs trap many larvae of a challenge infection within the liver. In this study the distribution of challenge larvae in sensitized mice was examined to determine the earliest onset of liver trapping and to establish if the previously described phenomenon truly represented larval trapping. In all experiments, C57BL/6J mice were infected with a sensitization dose of 125 infective T. canis eggs on day 0 postinfection (PI) and challenged with 500 infective eggs on day 28 PI. In the initial experiments, larval numbers were determined within the intestinal contents, intestinal wall, mesenteric tissues, liver, lungs, skeletal muscle, and brain of each mouse on days 0.5, 1, 2, 3, 5, and 6 postchallenge (PC). Migration patterns were similar among the test and control groups except the peak of larval numbers in the liver, seen at 1 day PC in control mice, was delayed until 3 days PC in the test group. Larval trapping occurred within the liver of test mice at least by day 5 PC. In subsequent experiments, larval numbers were determined within the liver, skeletal muscle, brain of each mouse, and within the eyes of each mouse group at 4, 8, 12, and 16 wk PC. Larval numbers within the liver of test mice were similar both at 5 days PC and 16 wk PC, implying that larvae were trapped in this organ rather than delayed in their migration to other body sites. Liver trapping did not protect the eyes or brain of sensitized mice from larval migration, nor did it result in larval killing.

Animals