Oral Ingestion of Yuzu Seed Oil Suppresses the Development of Atopic Dermatitis-like Skin Lesions in NC/Nga Mice.

Long-term oral ingestion of unheated yuzu seed oil in humans reduces lipid peroxides in the blood. Moreover, yuzu seed oil contains limonin, which can induce antioxidant and anti-inflammatory effects by activating the transcription factor nuclear factor erythroid 2-related factor 2 (Nrf2). Previously, Nrf2 has been shown to reduce atopic dermatitis (AD). Therefore, we hypothesized that ingesting unheated yuzu seed oil can regulate AD through Nrf2. An AD model was established using NC/Nga mice through repeated local exposure to mite antigens. Unheated and purified yuzu seed oil (100 µL/mice) or water (control, 100 µL/mice) was administered orally once a day using a gastric cannula for rodents for 28 days. On day 28, mice in the unheated yuzu seed oil group exhibited significantly lower clinical skin severity scores and ear thickness than those in the purified yuzu seed oil and water groups. Serum histamine levels remained unaltered among the three AD-induced groups. Serum Dermatophagoides farina body (Dfb)-specific immunoglobulin E (IgE) levels were significantly lower in the unheated yuzu seed oil group. Oral ingestion of yuzu seed oil in NC/Nga AD model mice significantly suppressed dermatitis deterioration and decreased serum IgE levels. Clinical trials (n = 41) have already confirmed that unheated yuzu oil is safe for long-term intake, further suggesting its potential use in improving AD symptoms.

Successful treatment of multiple microbleeds in a large area of the small bowel by transcatheter arterial embolization using imipenem/cilastatin as embolization material.

A 44-year-old man with chronic idiopathic pseudo-intestinal obstruction and lumbar disc herniation presented with orthostatic dizziness, black vomiting, and stools. He was suspected to have an ulcer caused by nonsteroidal anti-inflammatory drugs and treated conservatively but continued to have transfusion-dependent anemia. Trans-arterial contrast-enhanced computed tomography showed multiple microbleeds in the small intestine. We diffusely embolized 7 small intestine branches of the superior mesenteric artery using imipenem/cilastatin on 2 separate occasions. This stopped the bleeding, and the patient progressed well without ischemic complications and was discharged on the 25th postoperative day. Transcatheter arterial embolization with imipenem/cilastatin may be a viable treatment option for patients with multiple small bowel bleeds in a large area of the small intestine that are unresponsive to conservative treatment or endoscopic methods.

Transcatheter arterial embolization for acute lower gastrointestinal bleeding using imipenem/cilastatin: a single-center retrospective study.

BACKGROUND: Transcatheter arterial embolization (TAE) is a standard treatment for acute lower gastrointestinal bleeding (LGIB) in situations where endoscopic approaches are impossible or ineffective. Various embolic materials, such as metallic coils and N-butyl cyanoacrylate, are used. This study aimed to evaluate the clinical outcomes of an imipenem/cilastatin (IPM/CS) mixture as an embolic agent in TAE for acute LGIB. RESULTS: Twelve patients (mean age, 67 years) with LGIB treated with TAE using IPM/CS were retrospectively evaluated between February 2014 and September 2022. All patients showed evidence of extravasation on computed tomography and 50% (6/12) also showed evidence on angiography. The technical success rate for TAE in this study was 100%, including in patients who showed active extravasation on angiography. The clinical success rate was 83.3% (10/12), with two patients experiencing rebleeding within 24 h after the procedure. No ischemic complications were observed, and no bleeding episodes or other complications were reported during the follow-up period. CONCLUSIONS: This study revealed that using IPM/CS as an embolic agent in TAE for acute LGIB may be safe and effective, even in cases of active bleeding.

Imaging findings and complications of transcatheter interventional treatments via the inferior phrenic arteries in patients with hepatocellular carcinoma.

OBJECTIVE: To evaluate imaging findings and complications from transcatheter interventional treatment of hepatocellular carcinoma via the inferior phrenic arteries. MATERIAL & METHODS: 40 procedures in 25 patients (19 men; age range, 57-89 years) were retrospectively reviewed in this study. In all procedures, a micro-catheter was selectively inserted in the right inferior phrenic artery (n = 39) or left inferior phrenic artery (n = 1), and transcatheter arterial chemoembolization (n = 39) or transcatheter arterial embolization (n = 1) was performed. Imaging findings and patient charts were reviewed, and complications until time of discharge (median hospitalization period, 10.5 days; range, 3-21) were assessed. RESULTS: On angiography or computed tomography during angiography, collateral circulation from the right inferior phrenic artery to the pulmonary artery was seen in eight of 39 procedures (seven patients, 28%). In seven of these procedures, Lipiodol deposition was seen on the unenhanced computed tomography just after the procedure (post-procedure computed tomography) in the pulmonary arteries or pleura, and in six procedures, the deposited Lipiodol was noted to have spread into adjacent lung fields on the one week follow-up computed tomography. Branches of the right inferior phrenic artery were seen along the right margin of the heart in 18 procedures, and Lipiodol deposition was seen along the right margin of the heart on post-procedure computed tomography in four procedures. Complications occurred in 21 of 39 procedures of right inferior phrenic artery intervention (53%): shoulder pain in 18 (45%), pleural effusion in 14 (35%), basal atelectasis in 11 (28%), paroxysmal atrial fibrillation in two (5%) and hemoptysis in one (3%). In 14 procedures (35.9%), pleural effusion was seen on follow-up computed tomography examinations, and 11 (28.2%) of these procedures also showed basal atelectasis. However, only three procedures with pleural effusion showed Lipiodol deposition on the post-procedure computed tomography. In one patient who underwent transcatheter arterial chemoembolization twice via the right inferior phrenic artery, atrial fibrillation occurred after both procedures. CONCLUSIONS: Transcatheter arterial chemoembolization or transcatheter arterial embolization via the inferior phrenic artery in patients with hepatocellular carcinoma was relatively safe. Shoulder pain was the most frequent complication, and required only conservative treatment. There was no clear connection between pleural effusion or basal atelectasis and collateral circulation from the right inferior phrenic artery to the pulmonary artery depicted on angiography, computed tomography during angiography or post-procedure computed tomography.

The adjuvant effect of Sophy ß-glucan to the antibody response in poultry immunized by the avian influenza A H5N1 and H5N2 vaccines.

Avian influenza virus vaccines produced in oil-emulsified inactivated form with antigen content of at least 160 hemagglutinin units (HAU) induced immunity in birds. However, in addition to enhancing the effect of the adjuvant(s), other additional supplemented biological compounds included in inactivated vaccines could produce higher levels of antibody. We examined in chickens, Vietnamese ducks, and muscovy ducks the adjuvant effect of Sophy ß-glucan (SBG), a ß-1,3-1,6 glucan produced by the black yeast Aureobasidium pollulans strain AF0-202, when administered with an avian influenza H5 subtype vaccine. In Experiment 1, 40 chickens (ISA Brown hybrid), allocated to four groups of ten each, were immunized with Oil-H5N1(VN), Oil-H5N1(CN), Oil-H5N2(CN), and saline (control group), respectively. In Experiment 2, chickens (ISA Brown hybrid), muscovy ducks (French hybrid), and Vietnamese ducks (indigenous Vietnamese) were used to further assess the effect of SBG on immunogenicity of the Oil-H5N1(VN) Vietnamese vaccine. ELISA and hemagglutination inhibition (HI) assays were used to assess the antibody response. The H5 subtype vaccines initiated significantly higher immune responses in the animals dosed with SBG, with 1.0-1.5 log2 higher HI titers and 10-20% ELISA seroconversion, compared with those not dosed with ß-glucan. Notably, some of the animals dosed with SBG induced HI titers higher than 9.0 log2 following boosting immunization. Taken together, our serial studies indicated that SBG is a potential effector, such as enhancing the immune response to the H5 vaccines tested.

Effect of Sophy ß-glucan on immunity and growth performance in broiler chicken.

This study was carried out to determine the effect of Sophy ß-glucan on immunity and growth performance in broilers. One group was treated with 1% ß-glucan ad libitum with water and the other group was kept as the control. Vaccination for Infectious Bursal Disease was carried out on days 16 and 21. Blood samples were collected from birds, and antibody titres against IBD were measured. The mean body weight of the ß-glucan treated group was significantly (P<0.05) higher than that of the control group. The mean antibody titres measured on days 25, 36 and 42 were significantly (P<0.05) higher in 1% ß-glucan treated group than that of the control group, suggesting the presence of immune stimulating effect of ß-glucan.

Aureobasidium-derived soluble branched (1,3-1,6) beta-glucan (Sophy beta-glucan) enhances natural killer activity in Leishmania amazonensis-infected mice.

The beta-glucans derived from yeast cell walls have been reported for having many immunomodulatory activities in vivo and in vitro. In this study, Aureobasidium-derived soluble branched (1,3-1,6) beta-glucan (Sophy beta-glucan) was checked for natural killer (NK) activity and for the production of IFN-gamma and IL-4 in Leishmania amazonensis infection. The main experiment was performed with a group of female C57BL/6 and BALB/c mice, orally supplemented with 5% of Sophy beta-glucan and infected with promastogotes of L. amazonensis (1 x 10(7)) into the footpad. Increase in the footpad thickness with time was observed in BALB/c mice in spite of the oral Sophy beta-glucan supplement, but it was less in C57BL/6 mice. The difference in overall mean footpad thickness between 'infection only' versus 'infection + glucan' groups was statistically significant (P < 0.001). High NK activity in C57BL/6 than BALB/c mice was observed in 'glucan only' group compared to the control group and also in 'infection + glucan' group compared to 'infection only' group. The difference in the NK activity among these groups was significant (P < 0.05). The IFN-gamma level increased at weeks 7 and 8 post-infection in C57BL/6 mice and was significantly high in 'infection + glucan' group compared to the 'infection only' group (P < 0.05). IL-4 levels did not increase up to detectable levels throughout the study. The results led a conclusion that Sophy beta-glucan enhances NK activity and cellular immunity in L. amazonensis-infected mice.

Development of a highly sensitive IgG-ELISA based on recombinant arginine kinase of Toxocara canis for serodiagnosis of visceral larva migrans in the murine model.

Toxocariasis is a worldwide zoonotic disease caused by the ascarid nematode Toxocara canis. The most common method available for serodiagnosis of toxocariasis is an enzyme-linked immunosorbent assay (ELISA) test using Toxocara excretory-secretory antigen (TES). The present study describes the development of IgG-ELISA based on antiserum prepared against the recombinant arginine kinase of Toxocara canis. Antiserum was prepared against the purified recombinant arginine kinase (AK) using 6-week-old female Japanese white rabbits. Serum samples were collected from experimentally infected BALB/c and C57BL/6 mice at different time periods. The IgG-ELISA was performed using serum samples from mice (infected/uninfected) and TES antigen with antiserum prepared against the recombinant-AK. The optical density (OD450) was measured at 450 nm using a micro-plate ELISA reader. There were significant differences (P<0.01) in the absorbance between infected and control serum samples. Further, we obtained 100% sensitivity for the serum samples from T. canis-infected mice. Therefore, it is suggested that the recombinant-AK based IgG-ELISA could be applied for immunodiagnosis of human toxocariasis. However, it is necessary to evaluate the specificity of this recombinant antigen with similar geohelminth infections.

Toxocara canis: molecular cloning, characterization, expression and comparison of the kinetics of cDNA-derived arginine kinase.

Arginine kinase (AK) is a member of a highly conserved family of phosphagen kinases. We determined the cDNA sequence of Toxocara canis AK, cloned it in pMAL plasmid and expressed it in Escherichia coli as a fusion protein with maltose-binding protein. The protein has a theoretical molecular mass of 45,376 Da and an estimated isoelectric point (pI) of 8.38. Alignment of the cDNA-derived amino acid sequence of T. canis AK with other phosphagen kinase sequences showed high amino acid identity with other nematode AKs, and phylogenetic analysis placed it as a distinct branch within a nematode AK cluster. Analysis of the N-terminus sequence of T. canis AK revealed the presence of a signal targeting peptide presumably targeting this protein to cytosol or endoplasmic reticulum (ER). T. canis AK showed high activity for l-arginine. The kinetic constants (K(m) = 0.12 mM, K(cat) = 29.18, and K(d) = 0.23 mM) and V(max) (43.76 micromolPi/min/mg protein) of T. canis recombinant-AK were determined for the forward reaction. It also exhibited a synergism for substrate binding (K(d)(Arg)/K(m)(Arg)=1.96). Comparison of K(cat)/K(m)(Arg) values in various arginine kinases indicates that T. canis AK has a high catalytic efficiency (248.19s(-1)mM(-1)). The present study contains the first description of arginine kinase in a zoonotic nematode. The determination of T. canis AK and its phosphagen biosynthetic pathway, which is completely different from those in mammalian host tissues, suggests this enzyme as a possible novel chemotherapy target for VLM syndrome in humans.

IL-5-Induced Eosinophils Suppress the Growth of Leishmania amazonensis In Vivo and Kill Promastigotes In Vitro in Response to Either IL-4 or IFN-gamma.

In IL-5 transgenic mice (C3H/HeN-TgN(IL-5)-Imeg), in which 50% of peripheral blood leukocytes are eosinophils, the development of infection by Leishmania amazonensis was clearly suppressed. To determine mechanistically how this protozoan parasite is killed, we performed in vitro killing experiments. Either IL-4 or IFN-gamma effectively stimulated eosinophils to kill Leishmania amazonensis promastigotes, and most of the killing was inhibited by catalase but not by the NO inhibitor L-N5-(1-iminoethyl)-ornithine, suggesting that hydrogen peroxide is responsible for the killing of L. amazonensis by eosinophils. There was no significant degranulation of eosinophils in the culture, because eosinophil peroxidase was not detected in culture supernatants when L. amazonensis promastigotes were killed by activated eosinophils. Such resistance was also observed in BALB/c mice, which are highly susceptible to L. amazonensis. Expression plasmids for IL-4, IL-5, and IFN-gamma were transferred into muscle by electroporation in vivo starting 1 week before infection. Expression plasmid for IL-5 was most effective in slowing the development of infection among three expression plasmids. Expression plasmid for IL-4 was slightly effective and that for IFN-gamma had no effect on the progress of disease. These results suggest that IL-5 gene transfer into muscle by electroporation is useful as a supplementary protection method against L. amazonensis infection.

Mitochondrial transmembrane potential is diminished in phorbol myristate acetate-stimulated peritoneal resident macrophages isolated from wild-type mice, but not in those from gp91-phox-deficient mice.

Macrophages produce superoxide (O2-) during phagocytosis or upon stimulation with a variety of agents including phorbol myristate acetate (PMA) through the activation of NADPH oxidase, and the formed O2- is converted to other reactive oxygen species (ROS) such as hydrogen peroxide (H2O2). The aim of the present study was to elucidate the effect of the intracellularly produced ROS on mitochondrial transmembrane potential (MTP) in mouse (C57BL/6) peritoneal resident macrophages stimulated with PMA. Using a fluorescent dye, succinimidyl ester of dichlorodihydrofluorescein (H2DCFDA), O2- was visualized in intracellular compartments in a certain subpopulation of macrophages isolated from wild-type mice. Cells deficient in gp91-phox, one of the membrane components of NADPH oxidase, were negative for the fluorescence. When cells were loaded with both H2DCFDA and MitoCapture, a fluorescent dye for mitochondria, mitochondrial fluorescence was diminished in O2- -producing cells, but not in O2- -deficient cells. Flow cytometry also revealed the decrease of mitochondrial fluorescence in wild-type cells, but not in gp91-phox-deficient cells. The loss of mitochondrial fluorescence was prevented by microinjection of catalase into cells. The present findings demonstrate that MTP is diminished by ROS, including the H2O2 dismutated from O2-, produced intracellularly by activation of the NADPH oxidase in mouse peritoneal resident macrophages stimulated with PMA.