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PubMed2006

Inhibition of Escherichia coli O157:H7, Listeria monocytogenes and Staphylococcus aureus on sliced roast beef by cetylpyridinium chloride and acidified sodium chlorite.

The effects of 0.5% cetylpyridinium chloride (CPC), 0.12% acidified sodium chlorite (ASC) and a mix of equal volume of the two (0.25% CPC-0.06% ASC) on Escherichia coli O157:H7, Listeria monocytogenes and Staphylococcus aureus were evaluated on inoculated sliced roast beef. The antimicrobial agents were, respectively, sprayed on the beef surfaces and tray absorbent pads, and samples were stored at 4 degrees C for 10 days (d). At 0 d, L. monocytogenes and S. aureus were reduced to undetectable levels in 2 h after spraying with CPC. CPC-ASC treatment reduced E. coli O157:H7, L. monocytogenes and S. aureus by 4.07, 6.37 and 4.32 log cfu/cm2, respectively, at 0 d. ASC treatment reduced the population of E. coli O157:H7 by 6.09 log cfu/cm2 at 10 d. CPC treatment caused a slight discoloration and ASC-treated beef surfaces demonstrated the lowest redness and highest lightness. The grey colour and off-odour were significant in the ASC-treated beef samples, while CPC-treated samples demonstrated less off-odor and brown colour from 0 to 4 d. Based on our results, it appears that the application of CPC on sliced roast beef can extend the shelf-life of the product without impairing its quality.

Animals↗

PubMed2005

Growth factor deposition in anterior subcapsular cataract.

PURPOSE: To characterize immunohistochemically the distribution of growth factors and extracellular matrix (ECM) components in an anterior subcapsular cataract (ASC) and to determine the role of growth factors in the development of ASC. SETTING: Department of Ophthalmology, Wakayama Medical University, Wakayama, Japan. METHODS: During cataract surgery in 22 patients, anterior capsules with an ASC were obtained. Sections of each specimen were immunostained with a panel of antibodies against ECM components, growth factors, cytoskeletal components, and signal transduction-related molecules. RESULTS: Collagen types I, V, and VI; fibronectin; fibrillin-1; and latent transforming growth factor beta binding protein-1 (LTBP-1) were localized to the ECM in ASC tissues. Collagen IV was localized to the ECM and the capsule. Lens epithelial cells (LECs) were positive for alpha-smooth muscle actin (alphaSMA). Lens epithelial cells and ECM stained for transforming growth factor beta2 (TGFbeta2) and TGFbeta3 in all samples, but TGFbeta1 latency-associated peptide (TGFbeta1-LAP) were detected in some samples. Fibroblast growth factor-2 (FGF-2) and hepatocyte growth factor-alpha (HGF-alpha) were localized to the ECM. Lens epithelial cells with nuclear staining for Erk-1, the mitogen-activated protein kinase (MAP kinase) cascade-related molecule, and Smad3, 1 of the Smad family members involving TGFbeta signaling, were detected. CONCLUSIONS: Matrix components (ie, collagen types, fibronectin, fibrillin-1), as well as growth factors such as TGFbeta1-LAP, TGFbeta2, TGFbeta3, FGF-2, and HGF-alpha, were detected in ASC. Fibrillin-1 might serve as a repository for TGFbetas. These growth factors may modulate the phenotypic alteration and behavior of LECs. The MAP kinase cascade and TGFbeta signaling are both activated in LECs in ASC.

Adult↗

PubMed2005

Both direct and collagen-mediated signals are required for active vitamin D3-elicited differentiation of human osteoblastic cells: roles of osterix, an osteoblast-related transcription factor.

In order to investigate the mechanisms by which 1alpha,25(OH)2 vitamin D3 (VD3) stimulates the differentiation of human osteoblasts, we cultured MG-63, which is a human osteoblastic cell line, in the presence or absence of VD3 and/or L-ascorbic acid 2-phosphate (Asc 2-P), a long-acting vitamin C derivative. The cell growth rate was decreased by the presence of VD3 in the culture medium. Type I collagen synthesis and alkaline phosphatase (ALP) activity, which are markers of early stage osteoblast differentiation, were stimulated by the presence of VD3 as well as by that of Asc 2-P. The co-presence of Asc 2-P and VD3 had a synergistic effect on the collagen synthesis and ALP activity of the cells. Inhibition of collagen synthesis by the addition of inhibitors of collagen synthesis to the medium attenuated the stimulative effect of VD3 and Asc 2-P on the ALP activity. Transfection of the cells with siRNA-expressing vectors for COL1A1 decreased the expression level of ALP mRNA in addition to that of COL1A1. On the other hand, ALP activity was significantly increased, and the growth rate was decreased, when the cells were cultured on type I collagen-coated dishes. These effects were not seen when the cells were cultured on dishes coated with heat-denatured collagen. VD3 also increased the mRNA levels for Runx2 and osterix, which are transcription factors critical for osteoblast differentiation, as well as those of differentiation markers such as bone/liver/kidney type ALP, COL1A1, (the gene for the alpha1 chain of type I collagen), and osteocalcin, in the cells. Normal human osteoblasts and human bone marrow-derived mesenchymal stem cells (hBMSC) showed quite similar responses to VD3. These results indicate that VD3-stimulated gene expression of type I collagen and that mature type I collagen produced in the presence of Asc 2-P mediates at least a part of the stimulative effects of Asc 2-P and VD3 on the differentiation of these human osteoblastic cells. Levels of mRNAs for ALP and COL1A1 were increased, but the level of Runx2 was decreased, by the expression of osterix in MG-63 cells. These results also suggest that VD3 controls the growth and differentiation of human osteoblastic cells by regulating the gene expression of osteoblast-related transcription factors as well as that of type I collagen, and that the co-presence of both signals is essential for VD3 to express full activity toward the differentiation of human osteoblasts.

Alkaline Phosphatase↗

PubMed2006

Successful nursing student practicum in an ambulatory surgery center.

ALTHOUGH MOST ACADEMIC programs use hospital-based sites for perioperative clinical experiences, the exponential growth in the number of ambulatory surgery centers (ASCs) and the increased variety of procedures performed in these settings make ASCs excellent venues for student learning experiences. NURSING STUDENTS in one program in Florida have the opportunity to take a perioperative practicum designed to allow them to learn about all aspects of patient care and administrative functions in ASCs. THE ASC PRACTICUM EXPERIENCE is popular with students and staff members alike, and ASC administrators are seeking ways to increase the number of students in the program in an effort to attract new graduates to ASCs.

Ambulatory Surgical Procedures↗

PubMed2001

The structure of symptom report in asthma: a reevaluation.

OBJECTIVES: We studied the structure of symptom report in a sample of British asthma patients using the Asthma Symptom Checklist (ASC). METHODS: The ASC was administered to 193 patients, together with a questionnaire on demographics and asthma-related information and the Perceived Control of Asthma Questionnaire. RESULTS: Principal Component Analysis yielded evidence for a six-dimensional structure of the ASC, with positively correlated subscales for panic-fear, irritation, obstruction-dyspnea, obstruction-congestion, fatigue, and hyperventilation symptoms. Individual subscales showed good to excellent item characteristics and internal consistencies in individual subscales. Panic-fear and obstruction-congestion were related to self-reported duration of oral corticosteroid intake. Other ASC subscales were positively related to aspects of health care utilization, limitations in daily activity, and lower perceived control of asthma mainly in women. The dyspnea subscale of the ASC showed only little relationship with either of these variables. CONCLUSION: The ASC can reliably assess the multidimensional structure of asthma symptom report. Its subscales are associated with important aspects of asthma management behavior.

Administration, Oral↗

PubMed2001

Angelica sinensis modulates migration and proliferation of gastric epithelial cells.

A crude extract from Angelica sinensis (ASCE), which mainly consists of polysaccharides, prevents ethanol- or indomethacin-induced gastric mucosal damage and promotes ulcer healing. The aim of this study was to test the hypothesis that ASCE has a direct stimulating effect on gastric epithelial cells for wound healing. We found that ASCE significantly promoted the migration of epithelial cells over an artificial wound on the surface of an RGM-1 monolayer. The extract also stimulated DNA synthesis in a dose-dependent manner and concomitantly increased EGF mRNA expression. Co-incubation of ASCE with anti-EGF antibody reduced the speed of migration and the DNA synthesis, which however were still higher than the control without ASCE. These results strongly suggest that ASCE has a direct wound healing effect on gastric mucosa, and this is acting partially through an EGF-mediated pathway.

Animals↗

PubMed2003

Effect of polysaccharides from Angelica sinensis on gastric ulcer healing.

Our previous study showed that a crude extract from Angelica sinensis (ASCE), which mainly consisted of polysaccharides, significantly promoted migration and proliferation of normal gastric epithelial cells. These results strongly suggest that ASCE has a direct wound healing effect on gastric mucosa. However, there is no report concerning the effect of ASCE on gastric ulcer healing in animal models. In this study, we found that ASCE promoted ulcer healing. The area of the ulcer was reduced. This was accompanied with a significant increase in mucus synthesis when compared with the control. Angiogenesis was inhibited by the treatment of ASCE. Cell proliferation, ODC and EGFR protein expression was not affected in this process. Thus, the mechanism of how ASCE accelerates ulcer healing in addition to its effect on mucus synthesis remains to be investigated.

Angelica sinensis↗

PubMed2000

Cloning and characterization of a human brain Na(+)-independent transporter for small neutral amino acids that transports D-serine with high affinity.

We isolated a cDNA for the human homologue of system asc transporter Asc-1 from human brain. The encoded protein designated as hAsc-1 (human Asc-1) exhibited 91 % sequence identity to mouse Asc-1. Consistent with mouse Asc-1, hAsc-1 required 4F2 heavy chain for its functional expression in Xenopus oocytes. hAsc-1 exhibited the properties of amino acid transport system asc which transports small neutral amino acids in a Na(+)-independent manner. hAsc-1 transported D-serine at high affinity with a K(m) value of 22.8 microM. In brain, 2.0 kb mRNA was highly expressed. hAsc-1 gene was mapped to human chromosome 19, region q12-q13.1. Because of the high-affinity transport with the K(m) value close to the physiological concentration of D-serine, together with the high levels of expression in brain, hAsc-1 is proposed to play significant roles in the D-serine mobilization in brain.

Amino Acid Transport System y+↗

PubMed1997

Quantitation of hydroxyl radicals produced by radiation and copper-linked oxidation of ascorbate by 2-deoxy-D-ribose method.

We have established controlled conditions for studying the reaction of chemically and radiolytically produced hydroxyl radical (.OH) with 2-deoxy-D-ribose (2-DR). Ascorbate (ASC) or dithiothreitol (DTT) and cuprous or cupric ions were used to generate the OH-radical. The OH-radical was detected using the classical method of measuring the amount of thiobarbituric acid reactive products (TBARP) formed by .OH-mediated 2-DR degradation, but using sensitive fluorescent detection of the TBARP production to quantify the OH-radical. All experiments were performed with adequate O(2) concentrations. The copper reaction with ASC consumes O(2) in a manner that is strongly dependent on copper concentration, and less dependent on ascorbate concentration. For an independent check of the Cu2+ catalyzed ASC oxidation kinetics, the decay of ASC absorbency at 265 nm, as well as the increase of H(2)O(2) absorbency at approximately 240 nm, were also monitored. These spectral changes agree well with the O(2) consumption data. TBARP production from 2-DR incubated with a Cu2+-ASC mixture or gamma-irradiated were also compared. gamma-Irradiation of 2-DR solutions shows a dose and 2-DR concentration dependent increase of TBARP generation. Other electron donors, such as DTT, are more complicated in their mechanism of OH-radical production. Incubation of 2-DR with Cu2+-DTT mixtures shows a delay (approximately 50 min) before OH-radical generation is detected. Our results suggest that the Cu2+-ASC reaction can be used to mimic the effects of ionizing radiation with respect to OH-radical generation. The good reproducibility and relative simplicity of the 2-DR method with fluorescence detection indicates its usefulness for the quantitation of the OH-radical generated radiolytically or chemically in carefully controlled model systems.

Ascorbic Acid↗

PubMed2002

Comparative permeation studies for delta-aminolevulinic acid and its n-butylester through stratum corneum and artificial skin constructs.

The improvement of the permeation properties through excised human stratum corneum and artificial skin constructs (ASC) of delta-aminolevulinic-n-butylester (ABE) compared with delta-aminolevulinic acid (ALA) was investigated. For this purpose the permeated amounts of each substance were determined depending on time in a Franz diffusion cell experiment with stratum corneum and ASC, respectively. Furthermore the barrier properties of ASC were compared with those of stratum corneum. Detection of both substances was performed by high-performance liquid chromatography (HPLC) analysis. For the determination of ABE a new HPLC method was developed. ABE could be determined with the new HPLC method with sufficient sensitivity (detection limit: 0.1 microg/ml) after derivatisation with o-phthalaldehyde (OPA). Stratum corneum and ASC were more permeable for ABE than for ALA. The permeation coefficient P of ABE through stratum corneum was nearly ten-fold higher than that of ALA. Using ASC as permeation barrier the permeation coefficient of ABE was about 22-fold higher than that of ALA. ABE and ALA permeated 142-fold and 64-fold, respectively, faster through ASC than through stratum corneum.

Aminolevulinic Acid↗

PubMed2002

Effect of tibolone (Org OD14) and its metabolites on aromatase and estrone sulfatase activity in human breast adipose stromal cells and in MCF-7 and T47D breast cancer cells.

Tibolone (Org OD14) is a synthetic steroid used for post-menopausal hormone replacement therapy (HRT). Since HRT might increase breast cancer risk, it is important to determine the possible effects of tibolone on breast tissues. Tibolone and its metabolites Org 4094, Org 30126 and Org OM38 have been reported to inhibit estrone sulfatase activity in MCF-7 and T47D breast cancer cell lines, which suggest beneficial effects on hormone dependent breast cancer by reducing local production of free estrogens. Breast adipose stromal cells (ASCs) contain aromatase activity-an obligatory step in the biosynthesis of estrogens-and possibly contain sulfatase activity. We investigated the effects of tibolone, its metabolites and the pure progestin Org 2058 on PGE(2)-stimulated aromatase activity and on sulfatase activity in human ASC primary cultures and on sulfatase activity in MCF-7 and T47D cell lines. In MCF-7, tibolone and metabolites, but not Org 2058, were found to inhibit sulfatase activity. In T47D, tibolone inhibited sulfatase only at 10(-6)M, although weakly. ASC had high sulfatase activity, which was inhibited by 10(-6)M of tibolone, Org 4094 and Org 30126, but not by Org OM38 or Org 2058. Surprisingly, aromatase activity in ASC was increased by both tibolone and Org 2058 at 10(-6)M. As ligand binding assay results and immunohistochemistry indicated the absence of progesterone and estrogen receptors in ASC, these effects on aromatase and sulfatase activity in ASC likely take place by other routes. Because tibolone and its metabolites inhibit sulfatase activity, and because tibolone only increases aromatase activity at a high concentration, we conclude that effects of tibolone on the breast are probably safe.

17-Hydroxysteroid Dehydrogenases↗

PubMed2000

Internet-based monitoring and benchmarking in ambulatory surgery centers.

BACKGROUND: Each year the number of surgical procedures performed on an outpatient basis increases, yet relatively little is known about assessing and improving quality of care in ambulatory surgery. Conventional methods for evaluating outcomes, which are based on assessment of inpatient services, are inadequate in the rapidly changing, geographically dispersed field of ambulatory surgery. Internet-based systems for improving outcomes and establishing benchmarks may be feasible and timely. METHODS: Eleven freestanding ambulatory surgery centers (ASCs) reported process and outcome data for 3,966 outpatient surgical procedures to an outcomes monitoring system (OMS), during a demonstration period from April 1997 to April 1999. ASCs downloaded software and protocol manuals from the OMS Web site. Centers securely submitted clinical information on perioperative process and outcome measures and postoperative patient telephone interviews. Feedback to centers ranged from current and historical rates of surgical and postsurgical complications to patient satisfaction and the adequacy of postsurgical pain relief. RESULTS: ASCs were able to successfully implement the data collection protocols and transmit data to the OMS. Data security efforts were successful in preventing the transmission of patient identifiers. Feedback reports to ASCs were used to institute changes in ASC staffing, patient care, and patient education, as well as for accreditation and marketing. The demonstration also pointed out shortcomings in the OMS, such as the need to simplify hardware and software installation as well as data collection and transfer methods, which have been addressed in subsequent OMS versions. DISCUSSION: Internet-based benchmarking for geographically dispersed outpatient health care facilities, such as ASCs, is feasible and likely to play a major role in this effort.

Benchmarking↗

PubMed2002

Quantitative analysis of wild-type and precore mutant hepatitis B virus in carriers.

In the present study, we have analyzed the amount of precore wild-type hepatitis B virus (HBV) (wild-type) and precore mutant HBV (nt 1896: G-->A) (precore mutant) of HBV carriers; 31 asymptomatic healthy carriers (ASC) and 28 patients with chronic hepatitis (CH). Wild-type and precore mutant were quantified using sensitive and specific quantification methods: competitive wild-type-sequence-specific assay and competitive mutation-site-specific assay with different sets of specific primers and internal controls. Median serum levels of wild-type and precore mutant were 9.60 and 8.60 log copies/ml (median percentages of precore mutant in total HBV-DNA: 11.7%) in HBeAg(+) ASC, 8.48 and 8.00 (33.3%) in HBeAg(+) CH, and 6.30 and 6.85 (84.7%) in anti-HBe(+) CH, respectively, showing higher levels of the relative amount of precore mutant to wild-type along with HBeAg/anti-HBe status. Only precore mutant, but not wild-type was detected in anti-HBe(+) ASC. Although median percentages of precore mutant at the anti-HBe(+) ASC and CH stages were much higher than those at the HBeAg(+) ASC and CH stages, a substantial amount of precore mutant was found even at the HBeAg(+) stages. Existence of a substantial amount of precore mutant even in HBeAg(+) ASC suggests that the occurrence of precore mutant is not always closely associated with seroconversion from HBeAg to anti-HBe.

Journal Article↗

PubMed2003

Solid state coordination chemistry of oxomolybdenum organoarsonate materials.

The oxomolybdenum-arsonate system was investigated under hydrothermal conditions in the presence of charge-compensating copper(II)-organonitrogen complex cations as secondary building blocks. A series of materials of the Mo/O/As/Cu/ligand family has been prepared and structurally characterized. The architectures of the products reflect the identity of the arsonate component and the organonitrogen ligand, as well as the reaction conditions. The structural versatility of this emerging class of compounds is manifested by the one-dimensional structures of [[Cu(o-phen)(H(2)O)(2)](2)Mo(6)O(18)(O(3)AsOH)(2)] (1), [[Cu(terpy)](2)Mo(4)O(13)H(AsO(4))(2)].2H(2)O (2.2H(2)O), [[Cu(2,2'-bpy)(H(2)O)](2)Mo(6)O(18)(O(3)AsC(6)H(5))(2)].2H(2)O (4.2H(2)O), and [[Cu(o-phen)(H(2)O)](2)[Mo(6)O(18)(O(3)AsC(6)H(5))(2)]].4H(2)O (5.4H(2)O), by the two-dimensional materials [[Cu(2)(tpyprz)(H(2)O)(2)]Mo(6)O(18)(O(3)AsOH)(2)].2H(2)O (3.2H(2)O), [[Cu(terpy)](2)Mo(6)O(18)(O(3)AsC(6)H(5))(2)].H(2)O (6.H(2)O), and [[Cu(2)(tpyprz)]Mo(6)O(18)(O(3)AsC(6)H(5))(2)].2H(2)O (7.2H(2)O), and the molecular clusters [[Cu(2,2'-bpy)(2)](2)Mo(12)O(34)(O(3)AsC(6)H(5))(4)].2.35H(2)O (8.2.35H(2)O) and [Cu(o-phen)(H(2)O)(3)][Cu(o-phen)(2)Mo(12)O(34) (O(3)AsC(6)H(5))(4)].3H(2)O (9.3H(2)O).

Journal Article↗

PubMed2003

Antiproliferative and proapoptotic effect of ascorbyl stearate in human pancreatic cancer cells: association with decreased expression of insulin-like growth factor 1 receptor.

Pancreatic cancer is an aggressive tumor with short median survival and high mortality rate. Alternative therapeutic modalities are currently being evaluated for pancreatic cancer. Here we studied the effects of ascorbyl stearate (Asc-S), a nontoxic, lipophilic derivative of ascorbic acid, on pancreatic cancer. Treatment of human pancreatic carcinoma cells with Asc-S (50-200 microM) resulted in a dose-dependent inhibition of their proliferation. Asc-S slowed down the cell cycle, accumulating, PANC-1 cells in late G2-M phase. Furthermore, Asc-S treatment (150 microM) markedly inhibited growth in soft agar and facilitated apoptosis of PANC-1 cells but not of Capan-2 cells. These effects were accompanied by a significant reduction in insulin-like growth factor 1 receptor (IGF1-R) expression, as compared to untreated controls. Interestingly, Capan-2 cells, the least responsive to Asc-S treatment, did not overexpress the IGF1-R. The results demonstrate the efficacy of Asc-S in inhibing growth of pancreatic cancer cells and warrant additional studies to explore the potential utility of this compound as an alternative and/or adjuvant therapeutic modality for pancreatic cancer.

Apoptosis↗

PubMed2003

Mucosal and systemic immune responses to plasmid protein pgp3 in patients with genital and ocular Chlamydia trachomatis infection.

The circulating and cervical B cell responses to Chlamydia trachomatis plasmid protein pgp3 were characterized in children and adults with ocular or genital chlamydial infection using the enzyme-linked immunospot assay (ELISPOT) and ELISA. No pgp3-specific ASCs were detected in healthy controls, but predominantly IgA ASCs were detected in UK adults with uncomplicated cervicitis or urethritis (P = 0.03, 0.019). In patients with extragenital complications or pelvic inflammatory disease a mixed response with more IgG and IgM ASCs was evident, suggesting a breach of mucosal immune compartmentalization with more extensive infection. In women with chlamydial cervicitis, ASCs secreting predominantly IgA, but also IgG, to pgp3 were present in cervix at presentation, with a frequency 30-50 times higher than blood. Cervical ASC numbers, especially IgG, fell markedly six weeks after antibiotic treatment. We detected principally IgA pgp3-specific antibody secreting cells (ASCs) in children resident in a Gambian endemic area, with a trend towards suppression of IgA responses during intense trachomatous inflammation (P = 0.06), as previously reported for other chlamydial antigens, and in keeping with the findings in genital disease. These data provide a rationale for further studies of immune responses to pgp3 in humans and animal models of chlamydia-induced disease, and its potential use in diagnostic assays and protective immunization strategies.

Adult↗

PubMed2001

Short-term immunoglobulin A B-cell memory resides in intestinal lymphoid tissues but not in bone marrow of gnotobiotic pigs inoculated with Wa human rotavirus.

Immunological memory is important for protecting the host from reinfection. To investigate the development and sites of residence of intestinal memory B cells, and their role in protective immunity to reinfection with an enteric virus, we assessed the association between memory B cell and antibody-secreting cell (ASC) responses and protection using a gnotobiotic pig model for human rotavirus (HRV) infection and diarrhoea. The isotypes, quantities and tissue distribution of rotavirus-specific memory B cells and ASC were evaluated prechallenge (28 and 83 postinoculation days [PID]) and postchallenge (7 postchallenge days [PCD]), using enzyme-linked immunospot (ELISPOT) assay, in gnotobiotic pigs inoculated once with virulent or three times with attenuated HRV and challenged at PID 28 with the corresponding virulent HRV. Complete protection against HRV shedding and diarrhoea was associated with significantly higher numbers of immunoglobulin A (IgA) and immunoglobulin G (IgG) memory B cells and ASC in the ileum of virulent HRV-inoculated pigs at challenge. In contrast, pigs inoculated with attenuated HRV had lower numbers of IgA and IgG memory B cells and ASC in intestinal lymphoid tissues, but higher numbers in the spleen. The bone marrow had the lowest mean numbers of IgA and IgG memory B cells and ASC prechallenge in both groups of HRV-inoculated pigs. Therefore, bone marrow was not a site for IgA and IgG rotavirus-specific antibody production or for memory B cells after inoculation with live rotavirus, from 28 PID up to at least 83 PID. The effect of in vitro antigen dose was examined and it was determined to play an important role in the development of ASC from memory B cells for the different tissues examined.

Animals↗

PubMed2001

Regional immune responses with stage-specific antigen recognition profiles develop in lymph nodes of pigs following Ascaris suum larval migration.

The early life-cycle of the pig round worm, Ascaris suum, involves well-defined larval development in the liver, lungs and finally the small intestine. Distinct regional immune responses to larval antigens of A. suum were observed in the draining lymph nodes of immunized and challenged pigs during larval migration. This was reflected in a transient enlargement of the stimulated lymph nodes, due to increases in numbers of B cells and CD4 T cells, and the production of A. suum-specific antibody by antibody secreting cell (ASC) cultures. Larval antigen recognition pattern of antibodies in serum, bile and draining lymph node ASC culture supernatant (ASC-probes) was examined by immunoblotting. This revealed distinct organ-specific recognition patterns of larval-specific antigens by the draining lymph nodes at different times after challenge. In particular, an early larval 42 kDa antigen was recognized specifically by ASC-probes of the liver lymph nodes at 7 but not 14 days postchallenge (pc) which was not detected in other lymph nodes, serum or bile of the same pig. Similarly, a late larval antigen of 34 kDa was uniquely detected by lung and jejunal ASC-probes at 14 days pc. These observations demonstrate how development of distinct regional immune responses in tissues with different antigen stimulation can be monitored with ASC-probes and flow cytometry.

Animals↗