Intestinal Lymph Research Articles

The Impact of Lymphatic Transport on the Systemic Disposition of Lipophilic Drugs

This work investigates the influence of drug absorption route (intestinal lymphatics vs. blood supply) on drug pharmacokinetics and tissue distribution. To achieve this aim, the pharmacokinetics and tissue distribution of model compounds [1,1-bis(4-chlorophenyl)-2,2,2-trichloroethane, DDT; halofantrine] and lipids were assessed following intravenous delivery in lymph lipoproteins or plasma, and were found to differ significantly. For DDT, the clearance (CL) and volume of distribution (Vd ) were higher, whereas for halofantrine, CL and V(d) were lower, after entry in lymph versus plasma due, in particular, to differences in adipose tissue and liver uptake. In a recent study, halofantrine CL and V(d) were similar following entry in lymph or entry in plasma into the systemic circulation of animals predosed with lymph, whereas in the current study, predosing lymph did not influence DDT CL and V(d). For compounds such as DDT, changes to the route of absorption may thus directly impact on pharmacokinetics and tissue distribution, whereas for halofantrine factors that influence lymphatic transport may, by altering systemic lipoprotein concentrations, indirectly impact pharmacokinetics and tissue distribution. Ultimately, careful control of dosing conditions (formulation, prandial state), and thus the extent of lymphatic transport, may be important in assuring reproducible efficacy and toxicity for lymphatically transported drugs.

Early Trauma-Hemorrhage–Induced Splenic and Thymic Apoptosis Is Gut-Mediated and Toll-Like Receptor 4-Dependent

Immune depression after trauma-hemorrhage has been implicated as an important factor in the pathogenesis of sepsis and septic-organ failure. Although recent studies have implicated immune-cell apoptosis as an important factor in the evolution of this posttrauma immune-suppressed state, neither the initial triggers that induce this response nor the cellular pathways through which these triggering pathways act have been fully defined. Thus, the current study tests the hypothesis that acute splenic and thymic immune-cell apoptosis developing after trauma-hemorrhagic shock (T/HS) is due to gut-derived factors carried in intestinal lymph and that this T/HS lymph-induced immune depressed state is mediated through Toll-like receptor 4 (TLR4). The first set of experiments documented that T/HS caused both thymic and splenic immune-cell apoptosis as measured by TUNEL (terminal deoxynucleotidyl transferase dUTP nick end labeling) and caspase-3 immunohistochemistry and that this increase in apoptosis was totally abrogated by mesenteric lymph duct ligation. In subsequent experiments, mesenteric lymph collected from animals subjected to T/HS or trauma-sham shock were injected into TLR4-deficient (TLR4mut) mice or their wild-type (WT) littermates. Trauma-hemorrhagic shock, but not trauma-sham shock, lymph caused splenic apoptosis in the WT mice. However, the TLR4mut mice were resistant to T/HS lymph-induced splenic apoptosis. Furthermore, the WT, but not the TLR4mut mice developed splenic apoptosis after actual T/HS. In conclusion, gut-derived factors appear to initiate a sequence of events that leads to an acute increase in splenic and thymic immune-cell apoptosis, and this process is TLR4-dependent.

Ultrasonographic and general pathologic features assessment of small intestinal lymphoma

To study the ultrasonographic and morphologic features of intestinal lymphoma. The ultrasonographic and morphologic features of 19 patients with intestinal lymphoma were retrospectively analyzed. RESULTS All the patients were confirmed as non-Hodgkin's lymphoma by endoscopy or pathology. Of these 19 cases,the involved locations included ileum(n=14),duodenum(n=3),and ileocecum(n=2). The main sonographic types included mass type(n=12),bowel wall thickening type(n=1),and nonspecific signs(n=6). The mass type was characterized by solid and cystic-solid hypoechoic(including marked hypoechoic)foci,with well-defined margin,and rich blood signals were visible in large masses. The bowel wall thickening type was characterized by the thicking of bowel walls,showing hypoechoic(including marked hypoechoic),along with posterior acoustic enhancement. The nonspecific signs included e.g. dilatation of intestine and mesenteric lymph node enlargement. Morphologically,the tumor was featured by intestinal mass in intestinal mucosa or the circumferential thickening of intestinal wall. Small intestinal lymphoma has typical ultrasonographic features,and ultrasonography can provide useful information in the diagnosis of small intestinal lymphoma.

A unifying concept for periampullary adenocarcinomas

A unifying concept for periampullary adenocarcinomas

Dysregulated neutrophil function promotes chronic intestinal parasite infection (P3068)

Abstract Clearance of intestinal helminth infection and acquired resistance is dependent on an appropriate type-2 immune response. In the Trichuris muris (Tm) infection model in mice, resistant strains expel worms via a Th2 cell-mediated response whereas susceptible strains produce high levels of Th1 cell-associated cytokines and become chronically infected. Myeloid-cell specific deletion of Ship1 (SHIPΔLysM) in a normally resistant mouse strain (C57Bl6) results in chronic infection because of inappropriate production of IL12 in the intestine and mesenteric lymph nodes (mLNs) in Tm-infected mice. Although Ship1-deficient macrophages overproduce IL12 in vitro and in vivo and thus likely contribute to the Tm-susceptibility of SHIPΔLysM mice; surprisingly, deletion of Ship1 in the neutrophil lineage alone (SHIPΔPMN) is sufficient to render a resistant strain susceptible. Chronic worm burden in SHIPΔPMN mice is associated with elevated IL12, IFNγ and decreased IL13 production in the intestine and mLNs. These data suggest that regulation of neutrophil function by Ship1 is required for parasite clearance in response to Tm infection. Although neutrophils are rapidly recruited to the intestine following Tm infection, neutrophils are not required for clearance of parasites. However, our results suggest that dysregulated neutrophil activity can contribute to chronic parasitic infection.

Su1777 GIP Activation of Signaling Pathways Traffick PepT1 Proteins to the Apical Membrane of Intestinal Epithelial Cells

Background: In our previous study, using lymph fistula rat model, we have shown that fat ingestion stimulates release of mast cell mediators such as histamine, rat mucosal mast cell protease II (RMCP II), and prostaglandin D2 (PGD2) into intestinal lymph; however, it is unclear whether these responses are different depending on the type of fatty acid ingested. Aim: The purpose of this study was to determine the effects of dietary long-chain fatty acids with varying degrees of saturation on intestinal mast cell activation. Methods: We investigated lymphatic mast cell mediator concentrations after administration of three types of triacylglycerols, trilinolein (C18:2 n-6), trilinolenin (C18:3 n-3) or triolein (C18:1 n-9) by using the lymph fistula rat model. After cannulation of the major mesenteric lymph duct and duodenum of male Sprague-Dawley rat, a 3 ml bolus mixture of phosphate-buffered, taurocholatestabilized emulsion containing 120μmol of trilinolein or trilinolenin or triolein was provided to each animal through the intraduodenal feeding tube. The emulsion medium without triacylglycerol was used as control. Lymph was continuously collected for 6 hours and analyzed for RMCPII, histamine, PGD2, triglyceride and protein content. Results: Infusion of trilinolein and trilinolenin significantly increased the lymphatic release of mast cell mediators and influenced lymphatic protein concentration. In contrast, triolein did not induce the release of mast cell mediators. The peak RMCPII concentration after infusion of trilinolein (432.97 ± 77.53 ng/ml) and trilinolenin (317.50 ± 47.59 ng/ml) were significantly greater than triolein (141.46 ± 23.46 ng/ml) ( P,0.01, P,0.05, respectively). Similar to the RMCPII response, the peak values of histamine were significantly greater than triolein (12.97 ± 0.62ng/ml) for trilinolein (34.2 ± 1.7 ng/ml, P,0.01, n=3) and trilinolenin (19.85 ± 1.06ng/ ml, P,0.01, n=3). Infusion of trilinolein induced a significant increase of lymphatic PGD2 concentration peaked at 2 hour compared with the control (1461.23 ± 133.12 vs. 842.30 ± 129.02 pg/ml, P,0.05), but triolein did not induce (978.43 ± 153.74 pg/ml). Conclusion: These observations identify that the intestinal mast cell activation induced by dietary fat is dependent on the degree of unsaturation in fatty acid.

Su1776 Activation of Rat Intestinal Mast Cells Depends on Types of Unsaturated Long-Chain Fatty Acids

Background: In our previous study, using lymph fistula rat model, we have shown that fat ingestion stimulates release of mast cell mediators such as histamine, rat mucosal mast cell protease II (RMCP II), and prostaglandin D2 (PGD2) into intestinal lymph; however, it is unclear whether these responses are different depending on the type of fatty acid ingested. Aim: The purpose of this study was to determine the effects of dietary long-chain fatty acids with varying degrees of saturation on intestinal mast cell activation. Methods: We investigated lymphatic mast cell mediator concentrations after administration of three types of triacylglycerols, trilinolein (C18:2 n-6), trilinolenin (C18:3 n-3) or triolein (C18:1 n-9) by using the lymph fistula rat model. After cannulation of the major mesenteric lymph duct and duodenum of male Sprague-Dawley rat, a 3 ml bolus mixture of phosphate-buffered, taurocholatestabilized emulsion containing 120μmol of trilinolein or trilinolenin or triolein was provided to each animal through the intraduodenal feeding tube. The emulsion medium without triacylglycerol was used as control. Lymph was continuously collected for 6 hours and analyzed for RMCPII, histamine, PGD2, triglyceride and protein content. Results: Infusion of trilinolein and trilinolenin significantly increased the lymphatic release of mast cell mediators and influenced lymphatic protein concentration. In contrast, triolein did not induce the release of mast cell mediators. The peak RMCPII concentration after infusion of trilinolein (432.97 ± 77.53 ng/ml) and trilinolenin (317.50 ± 47.59 ng/ml) were significantly greater than triolein (141.46 ± 23.46 ng/ml) ( P,0.01, P,0.05, respectively). Similar to the RMCPII response, the peak values of histamine were significantly greater than triolein (12.97 ± 0.62ng/ml) for trilinolein (34.2 ± 1.7 ng/ml, P,0.01, n=3) and trilinolenin (19.85 ± 1.06ng/ ml, P,0.01, n=3). Infusion of trilinolein induced a significant increase of lymphatic PGD2 concentration peaked at 2 hour compared with the control (1461.23 ± 133.12 vs. 842.30 ± 129.02 pg/ml, P,0.05), but triolein did not induce (978.43 ± 153.74 pg/ml). Conclusion: These observations identify that the intestinal mast cell activation induced by dietary fat is dependent on the degree of unsaturation in fatty acid.

MULTICENTRIC LYMPHOMA IN A GIANT ANTEATER (MYRMECOPHAGA TRIDACTYLA)

Neoplastic disease is not well documented in giant anteaters. This report describes a disseminated lymphoma in an adult male giant anteater (Myrmecophaga tridactyla) from the City Zoo of Curitiba, State of Paraná, Brazil. No clinical signs were noticed before its death, except for a slight inappetence. At postmortem examination, pale white to yellow, variably sized nodules infiltrated the heart, liver, and intestinal lymph nodes. Histologically, two distinct cell populations were present in the nodular lesions: one characterized by smaller cells, primarily lymphocytic in nature, and another characterized by larger rounded cells with loose chromatin and frequently indented nuclei resembling histiocytes. Giant binucleated cells were occasionally observed. Mitotic figures numbered 2-3 mitotic figures/x400 field. Both cellular populations presented with moderate pleomorphism, large nuclei, a high nucleus-to-cytoplasm ratio, distinct nucleoli, and coarse nuclear chromatin. The neoplasia was classified as a form of multicentric lymphohistiocytic lymphoma (Rappaport Classification) and as an intermediate grade lymphoma (National Cancer Institute Working Formulation).

Study the Toll-like Receptor 2 and Dectin-1 in mice treated with beta‐ glucans and infected with Salmonella Typhimurium

Objective: Toll-like receptors (TLRs) are essential components for the induction of innate immune responses in different tissues including the small intestine, spleen and mesenteric lymph nodes. We investigated the expression of TLR2 in these tissues in mice infected with Salmonella Typhimurium. In this study we have examined how dectin-1, a lectin family receptor for β-glucans, collaborates with TLR-2 in recognition of microbes.Methods :In this study two groups of mice were treated with the β- glucan (local and commercial extract) and infected with(1x10 6 CFU/ml) S. Typhimurium at different intervals group 1 at day zero and group 2 after seven days of the experiment .The immunohistochemistory assay has been performed to detect TLR-2 and dectine-1 in the tissues of small intestine, spleen and mesenteric lymph nodes of the mice; the principle ofthis assay was done according to the manufacturer instructions (U.S.biological) .Results: In the immunohistochemistory assay, positive cells were indicated by cytoplasmic staining and the percentage of positive in 100 cells was recorded and graded. The grades were 1+ (<25%), 2+ (25-75%) and 3+ (>75%). Statistical analysis with Chi- Square tests revealed that there were significant differences (p˂0.05) between groups, also between grades within the same group in the small intestine, spleen with TLR2 expression; also we demonstrated the same results with Dectin-1 expression in the small intestine and mesenteric lymph nodes, except in the spleen which showed that there were no any relevant differences (p˂ 0.05 ) between the groups and the cell- grades.

Lactobacillus reuteri DSM 17938 Changes the Frequency of Foxp3+ Regulatory T Cells in the Intestine and Mesenteric Lymph Node in Experimental Necrotizing Enterocolitis

Necrotizing enterocolitis (NEC) is an inflammatory disease of the intestine in premature infants. Lactobacillus reuteri DSM 17938 improves survival and reduces the incidence and severity of NEC in a rodent model. Foxp3+ regulatory T cells (Tregs) maintain intestinal homeostasis by controlling inflammation and inducing tolerance. To determine whether there are insufficient numbers of Tregs to control inflammation in NEC and to determine if LR17938 increases the frequency of Tregs, we studied selected groups of newborn Sprague-Dawley rats according to feeding plan: dam±LR17938, formula±LR17938, and NEC±LR17938. NEC was induced by gavage feeding with special formula and exposure to hypoxic conditions. Lymphocytes isolated from ileum, mesenteric lymph nodes (MLN), spleen and thymus were labeled for T cell surface markers (CD3, CD4, CD8) and intracellular Foxp3; and labeled cells were analyzed by flow cytometry. The percentage of CD3+ T cells and Foxp3+ Tregs in the ileum significantly decreased in pups with NEC, compared to normal controls. Feeding LR17938 to neonatal rats with NEC increased the % of Foxp3+ T cells in the ileum while decreasing the percentage of cells in the MLN. Administration of LR17938 to dam-fed rats significantly increased Foxp3+Tregs in the ileum as early as day of life (DOL)1 but did not produce an increase in Tregs in formula-fed rats on DOL1. These results suggest that factors in breast milk may enhance the early immunomodulatory effects of LR17938. An anti-inflammatory effect of LR17938 in NEC was associated with the modulation of immune responses and induction and what appears to be migration of Foxp3+ Tregs to the diseased gut. Probiotic-facilitated development of Tregs might play an important role in the prevention of NEC.

Lymphatic diamine oxidase secretion stimulated by fat absorption is linked with histamine release

Diamine oxidase (DAO) is abundantly expressed in mammalian small intestine catalyzing the oxidative breakdown of polyamines and histamine. The aim of this study was to determine the relationship between stimulation of intestinal diamine oxidase secretion with intestinal fat absorption and histamine release. Conscious intestinal lymph fistula rats were used. The mesenteric lymph ducts were cannulated and intraduodenal tubes were installed for the infusion of Liposyn II 20% (an intralipid emulsion). Lymphatic DAO activity and protein secretion were analyzed by radiometric assay and Western blot, respectively. Lymphatic histamine concentration was measured by ELISA. Infusion of Liposyn II (4.43 kcal/3 ml) resulted in a ~3.5-fold increase in lymphatic DAO protein secretion and DAO activity, peaking at 1 h and lasting for 3 h. Liposyn II infusion also increased the lymphatic histamine release, a substrate for DAO. To determine the relationship of DAO release with histamine release, histamine was administered intraperitoneally (10 mg/kg) in fasting rats and resulted in a significant doubling in lymphatic DAO activity, supporting a link between histamine and DAO. In addition, ip administration of the histamine H4 receptor antagonist JNJ7777120 significantly reduced the Liposyn II-induced DAO output by 65.9%, whereas H(1) (pyrilamine maleate), H(2) (ranitidine), and H(3) (thioperamide maleate) receptor antagonists had little effect. We conclude that DAO secretion may contribute to the catabolism of histamine released during fat absorption and this is probably mediated through the histamine H(4) receptor.

The presence of Brucella ceti ST26 in a striped dolphin (Stenella coeruleoalba) with meningoencephalitis from the Mediterranean Sea

Brucella spp. was isolated from brain, lung and intestinal lymph nodes of a dead adult male striped dolphin (Stenella coeruleoalba) found stranded on the Tyrrhenian coast (Tuscany, Italy) of the Mediterranean Sea in February 2012. Brucella spp. was associated with moderate to severe lesions of meningoencephalitis. A co-infection by Toxoplasma gondii was also demonstrated at brain level by means of molecular and histopathologic methods. The Brucella isolate was further characterized based on a fragment-specific polymerase chain reaction (PCR) approach, consisting of a set of five specific PCRs, targeting specific chromosomal IS711 locations for marine mammal Brucellae, as described previously. The isolate was thus classified as Brucella ceti I; V fragment-positive (or B. ceti dolphin type), according to previous studies. Multi Locus Sequence Analysis demonstrated that the isolate belongs to Sequence Type 26, while omp2 (omp2a and omp2b genes) sequence analysis further confirmed the isolate belonged to this group of strains. This is the first report of Brucella spp. from marine mammals in the Mediterranean Sea, and represents a further observation that this strain group is associated with hosts of the Family Delphinidae, and particularly with the striped dolphins, also in the Mediterranean area, thus constituting a further biological hazard of concern for this vulnerable subpopulation.

Primary central nervous system histiocytic sarcoma presenting as a postradiation sarcoma: case report and literature review

Histiocytic sarcoma (HS) is a rare neoplasm that occurs most commonly in the intestinal tract, skin, soft tissue, and lymph node. The incidence of primary central nervous system (CNS) HS is even rarer, with a total of 6 cases reported in the literature. An etiologic link has not been identified for CNS HS, and the current case of primary CNS HS is unique in that an etiologic link to prior radiation therapy is identified, associated with complex cytogenetic abnormalities in the tumor. Although radiation-associated sarcomas can present as any number of different pathologic entities, this is the first reported case of a radiation-associated CNS HS. The pathologic and immunophenotypic characteristics of this case, with a nearly obscuring heavy inflammatory infiltrate and expression of monocytic/histiocytic markers (CD163, CD68, CD4, fascin), are characteristic of CNS HS. A discussion of the differential diagnosis and review of relevant literature are presented.

Primary intestinal lymphangiectasia diagnosed by double-balloon enteroscopy and treated by medium-chain triglycerides: a case report

IntroductionPrimary intestinal lymphangiectasia is a disorder characterized by exudative enteropathy resulting from morphologic abnormalities of the intestinal lymphatics. Intestinal lymphangiectasia can be primary or secondary, so the diagnosis of primary intestinal lymphangiectasia must first exclude the possibility of secondary intestinal lymphangiectasia. A double-balloon enteroscopy and biopsy, as well as the pathology can be used to confirm the diagnosis of intestinal lymphangiectasia. A polymeric diet containing medium-chain triglycerides and total parenteral nutrition may be a useful therapy.Case presentationA 17-year-old girl of Mongoloid ethnicity was admitted to our hospital with a history of diarrhea and edema. She was diagnosed with protein-losing enteropathy caused by intestinal lymphangiectasia. This was confirmed by a double-balloon enteroscopy and multi-dot biopsy. After treatment with total parenteral nutrition in hospital, which was followed by a low-fat and medium-chain triglyceride diet at home, she was totally relieved of her symptoms.ConclusionIntestinal lymphangiectasia can be diagnosed with a double-balloon enteroscopy and multi-dot biopsy, as well as the pathology of small intestinal tissue showing edema of the submucosa and lymphangiectasia. Because intestinal lymphangiectasia can be primary or secondary, the diagnosis of primary intestinal lymphangiectasia must first exclude the possibility of secondary intestinal lymphangiectasia. A positive clinical response to the special diet therapy, namely a low-fat and medium-chain triglyceride diet, can further confirm the diagnosis of primary intestinal lymphangiectasia.

High Fat and High Cholesterol Diet Induces DPP-IV Activity in Intestinal Lymph

Recent studies have reported that dipeptidyl-peptitase IV (DPP-IV) is correlated with diabetic conditions and also with dyslipidemia caused by overnutrition, especially a high fat diet. However, the role of DPP-IV in diabetes during dyslipidemia has been unclear. We utilized a lymph fistula rat model to determine whether intestinal lymph, which absorbs dietary fats, is affected by a chronic high-fat and high-cholesterol diet (HFHC). HFHC diet rats showed significantly higher DPP-IV activity in intestinal lymph and plasma compared to rats receiving a normal chow diet. In addition, HFHC diet rats showed significantly increased DPP-IV mRNA expression in the intestine. However, DPP-IV mRNA in the lymphocytes isolated from intestinal lymph and mesenteric lymph nodes did not show significant differences from that in the normal diet rats. In conclusion, HFHC diets increased DPP-IV expression in intestinal lymph; these results indicate the applicability of a previously unrecognized role for DPP-IV in metabolic disorders, including diabetes.

Genetic similarity of circulating and small intestinal virus at the end stage of acute pathogenic simian-human immunodeficiency virus infection

To understand the pathogenicity of acquired immune deficiency syndrome (AIDS), it is important to clarify where, when and how the virus replicates in the body of infected individuals. To identify the major virus replication site at the end stage of SHIV infection, we investigated the systemic tissues of SHIV-infected monkeys that developed AIDS-like disease. We quantified proviral DNA, and compared the mutation patterns of the viruses in various systemic tissues and in peripheral blood through phylogenetic analysis of the full genome sequence. We found that the amounts of proviral DNA detected in internal tissues were higher than those in peripheral blood mononuclear cells. In the sequence and phylogenetic tree analyses, the mutation patterns of the viruses in each tissue were generally different. However, the mutation pattern of the viruses in the jejunum and mesenteric lymph node were most similar to that of plasma viral RNA among the tissues examined in all three monkeys. In two of the three monkeys, which were euthanized earlier, viruses in the jejunum and mesenteric lymph node occupied the root position of the phylogenetic tree. Furthermore, in these tissues, more than 50% of SHIV-expressing cells were identified as macrophages based on co-expression of CD68. These results suggest that macrophages of the small intestine and/or mesenteric lymph node are the major virus production site at the end stage of SHIV infection of macaques.

The Effects of Erythropoietin on Bacterial Translocation and Inflammatory Response in an Experimental Intestinal Obstruction Model in Rats / Uticaj Eritropoetina Na Bakterijsku Translokaciju I Inflamatorni Odgovor U Eksperimentalnom Modelu Intestinalne Opstrukcije Kod Pacova

SummaryBackground:Intestinal obstruction results in distortion of balance of antiinflammatory cytokines and release of oxidants, and also leads to bacterial translocation, sepsis and multiple organ failure. Asymmetric dimethylarginine is related to multiple organ failure as a new prognostic marker. Erythropoietin reduces the inflammatory response by decreasing the levels of proinflammatory cytokines and cytokine-induced apoptosis. In this study, we aimed to investigate the effectiveness of erythropoietin in reducing the severity of bacterial translocation and inflammatory response after intestinal obstruction and the relation between asymmetric dimethylarginine and inflammatory markers.Methods:Forty Wistar albino rats (200-250 g) were divided into 4 groups as follows: Group 1 (Sham), only ileocaecal junction dissection; Group 2 (Erythropoietin), ileocaecal junction dissection and 3000 lU/kg erythropoietin subcutaneously; Group 3 (Intestinal Obstruction), complete ileal ligation; Group 4 (Intestinal Obstruction + Erythropoietin), complete ileal ligation and 3000 IU/kg erythropoietin subcutaneously. After 24 hours, the rats were sacrificed by taking blood from the heart for biochemical analyses. Peritoneal swab culture, liver, mesenteric lymph nodes, spleen and ileum were collected for microbiological and histopathological examinations.Results:Erythropoietin reduced the secretion of inflammatory cytokines, oxidative damage and bacterial translocation, prevented the formation of inflammatory changes in the intestine, liver, spleen and mesenteric lymph nodes,and also significantly prevented the formation of intestinal damage after intestinal obstruction (p<0.05). Conclusions: Asymmetric dimethylarginine levels did not differ between the groups. Erythropoietin may be useful to preserve from intestinal injury and related sepsis in patients with intestinal obstruction. Asymmetric dimethylarginine is not a suitable prognostic marker.

Biodistribution and Molecular Studies on Orally Administered Nanoparticle-AON Complexes Encapsulated with Alginate Aiming at Inducing Dystrophin Rescue inmdxMice

We have previously demonstrated that intraperitoneal injections of 2′-O-methyl-phosphorothioate (2′OMePS) antisense oligoribonucleotides adsorbed onto a cationic core-shell nanoparticles (NPs), termed ZM2, provoke dystrophin restoration in the muscles of mdx mice. The aim of the present work was to evaluate the oral route as an alternative way of administration for ZM2-antisense oligoribonucleotides complexes. The biodistribution and elimination of nanoparticles were evaluated after single and multiple oral doses of IR-dye conjugated nanoparticles. Labeled nanoparticles were tracked in vivo as well as in tissue cryosections, urines and feces by Odyssey infrared imaging system, and revealed a permanence in the intestine and abdominal lymph nodes for 72 hours to 7 days before being eliminated. We subsequently tested alginate-free and alginate-encapsulated ZM2-antisense oligoribonucleotides (AON) complexes orally administered 2 and 3 times per week, respectively, in mdx mice for a total of 12 weeks. Treatment with alginate ZM2-AON induced a slight dystrophin rescue in diaphragm and intestine smooth muscles, while no dystrophin was detected in alginate-free ZM2-AON treated mice. These data encourage further experiments on oral administration testing of NP and AON complexes, possibly translatable in oligoribonucleotides-mediated molecular therapies.

Prevalence of Swine Viral and Bacterial Pathogens in Rodents and Stray Cats Captured around Pig Farms in Korea

ABSTRACTIn 2008, 102 rodents and 24 stray cats from the areas around 9 pig farms in northeast South Korea were used to determine the prevalence of the following selected swine pathogens: ten viral pathogens [porcine epidemic diarrhea virus (PEDV), transmissible gastroenteritis virus (TGEV), rotavirus, classical swine fever virus (CSFV), porcine circovirus type 2 (PCV2), encephalomyocarditis virus (EMCV), porcine reproductive and respiratory syndrome virus (PRRSV), porcine parvovirus (PPV), pseudorabies virus (PRV) and Japanese encephalitis virus (JEV)] and four bacterial pathogens (Brucella, Leptospira, Salmonella and Lawsonia intracellularis). In total, 1,260 tissue samples from 102 rodents and 24 stray cats were examined by specific PCR and RT-PCR assays, including tissue samples of the brain, tonsils, lungs, heart, liver, kidneys, spleen, small intestine, large intestine and mesenteric lymph nodes. The percentages of PCR-positive rodents for the porcine pathogens were as follows: 63.7% for Leptospira, 39.2% for Brucella, 6.8% for Salmonella, 15.7% for L. intracellularis, 14.7% for PCV2 and 3.9% for EMCV. The percentages of PCR-positive stray cats for the swine pathogens were as follows: 62.5% for Leptospira, 25% for Brucella, 12.5% for Salmonella, 12.5% for L. intracellularis and 4.2% for PEDV. These results may be helpful for developing control measures to prevent the spread of infectious diseases of pigs.

The Effect of Plant Tissue and Vaccine Formulation on the Oral Immunogenicity of a Model Plant-Made Antigen in Sheep

Antigen-specific antibody responses against a model antigen (the B subunit of the heat labile toxin of enterotoxigenic Escherichia coli, LTB) were studied in sheep following oral immunisation with plant-made and delivered vaccines. Delivery from a root-based vehicle resulted in antigen-specific immune responses in mucosal secretions of the abomasum and small intestine and mesenteric lymph nodes. Immune responses from the corresponding leaf-based vaccine were more robust and included stimulation of antigen-specific antibodies in mucosal secretions of the abomasum. These findings suggest that oral delivery of a plant bioencapsulated antigen can survive passage through the rumen to elicit mucosal and systemic immune responses in sheep. Moreover, the plant tissue used as the vaccine delivery vehicle affects the magnitude of these responses.