• Parasites, Hosts and Diseases
• /
• v.54 no.1
• /
• pp.21-29
• /
• 2016

The sodium hydrogen exchanger 1 (NHE1), which functions in maintaining the ratio of $Na^+$ and $H^+$ ions, is widely distributed in cell plasma membranes. It plays a prominent role in pH balancing, cell proliferation, differentiation, adhesion, and migration. However, its exact subcellular location and biological functions in Toxoplasma gondii are largely unclear. In this study, we cloned the C-terminal sequence of T. gondii NHE1 (TgNHE1) incorporating the C-terminal peptide of NHE1 (C-NHE1) into the pGEX4T-1 expression plasmid. The peptide sequence was predicted to have good antigenicity based on the information obtained from an immune epitope database. After induction of heterologous gene expression with isopropyl-b-D-thiogalactoside, the recombinant C-NHE1 protein successfully expressed in a soluble form was purified by glutathione sepharose beads as an immunogen for production of a rabbit polyclonal antiserum. The specificity of this antiserum was confirmed by western blotting and immunofluorescence. The antiserum could reduce T. gondii invasion into host cells, indicated by the decreased TgNHE1 expression in T. gondii parasites that were pre-incubated with antiserum in the process of cell entry. Furthermore, the antiserum reduced the virulence of T. gondii parasites to host cells in vitro, possibly by blocking the release of $Ca^{2+}$. In this regard, this antiserum has potential to be a valuable tool for further studies of TgNHE1.

• Asian-Australasian Journal of Animal Sciences
• /
• v.20 no.11
• /
• pp.1670-1676
• /
• 2007

Fifty repeat breeder (RB) Friesian cows were allocated to five groups of 10 cows each, to determine the effect of progesterone (P4) supplement on P4 concentrations and pregnancy rates during the periods of corpus luteum (CL) formation and development between days 2-7 and 7-12 following a spontaneous or $PGF_{2{\alpha}}$-induced estrus. Cows were artificially inseminated during $PGF_{2{\alpha}}$-induced (PGF-P4-d2 and PGF-P4-d7 groups) or spontaneous (S-P4-d2, S-P4-d7, and control groups) estrus. Progesterone-releasing intravaginal device (PRID) devoid of estrogen capsule were inserted either on d 2 (PGF-P4-d2 and S-P4-d2 groups) or d 7 (PGF-P4-d7 and S-P4-d7 groups) post-insemination and left in place for 5 days. Control cows did not receive any treatment. Blood samples were collected for progesterone analysis from all cows once daily for 4 days starting on the day of estrus (d 0) and once every 3 days thereafter until d 22. Progesterone treatment by day interaction accounted for higher plasma P4 in treated than non-treated control cows. Progesterone concentrations differed significantly (p<0.05) during metestrus (d 2 to d 7) but not during diestrus (d 7 to d 12). $PGF_{2{\alpha}}$ treatment, lactation number, service number or their interactions did not affect progesterone concentrations and pregnancy rates. Therefore, cows were grouped according to the day of P4 supplement irrespective of the $PGF_{2{\alpha}}$ treatment. Progesterone supplement on d 7 but not d 2 significantly increased (p<0.03) pregnancy rates in repeat breeding cows with four or more previous services but not in cows in their third service. In conclusion, post-insemination P4 supplement to repeat breeding cows with four or more previous services improved pregnancy rates and should be advocated when no specific reason for infertility is diagnosed. Further studies with larger numbers of repeat breeding cows under field conditions are needed to ascertain the findings of this study.

• Journal of the Korea Society of Computer and Information
• /
• v.25 no.3
• /
• pp.185-190
• /
• 2020

The purpose of this study was to investigate the effects of systemic chronic inflammatory state on dry eye syndrome as a local chronic inflammatory condition. From June 16, 2016 to December 31, 2016, 726 patients who visited the general health examination center were screened using the Schirmer's test and the metabolic syndrome screening test. A total of 214 patients who were studied for hs-CRP and HbA1c were simultaneously selected for the study as well. Dry eye syndrome significantly increased in higher age groups (p<0.001) and women (p=0.020); there was no significant relationship with fasting plasma glucose, hs-CRP. In addition, as HbA1c increased the risk for dry eye syndrome also increased (β=1.960, p=0.025). Therefore, dry eye syndrome may not caused by microvascular changes in the lacrimal gland due to chronic inflammatory conditions. In diabetic patients, long-term blood glucose control may be more important than short-term blood glucose control. The high prevalence of dry eye syndrome in diabetic patients is thought to be due to autonomic dysfunction rather than microvascular changes caused by chronic inflammation.

• IMMUNE NETWORK
• /
• v.4 no.4
• /
• pp.216-223
• /
• 2004

Background: It is well known that IgA isotype switching is induced by $TGF-{\beta}1$. LPS-activated mouse normal B cells well differentiate into IgA secreting plasma cells under the influence of $TGF-{\beta}1$. Nevertheless, there are lots of difficulties in studying normal B cells in detail because it is not simple to obtain highly purified B cells, showing low reproducibility and transfection efficacy, moreover impossible to keep continuous culture. To overcome these obstacles, it is desperately needed to develop B cell line which acts like normal B cells. In the present study, we investigated whether CH12F3-2A lymphoma cells are appropriate for studying IgA isotype switching event. Methods: CH12F3-2A B cell line was treated with LPS and $TGF-{\beta}1$, then levels of germ-line (GL) transcripts were measured by RT-PCR, and $GL{\alpha}$ promoter activity was measured by luciferase assay. In addition, membrane IgA (mIgA) expression and IgA secretion were determined by FACS and ELISA, respectively. Results: $TGF-{\beta}1$, regardless of the presence of LPS, increased level of $GL{\alpha}$ transcripts but not $GL{\gamma}2b$ transcripts. However, IgA secretion was increased dramatically by co-stimulation of LPS and $TGF-{\beta}1$. Both mIgA and IgA secretion in the presence of $TGF-{\beta}1$ were further increased by over-expression of Smad3/4. Finally, $GL{\alpha}$ promoter activity was increased by $TGF-{\beta}1$. Conclusion: CH12F3-2A cell line acts quite similarly to the normal B cells which have been previously reported regarding IgA expression. Thus, CH12F3-2A lymphoma cell line appears to be adequate for the investigation of the mechanism(s) of IgA isotype switching at the cellular and molecular levels.

• IMMUNE NETWORK
• /
• v.14 no.1
• /
• pp.21-29
• /
• 2014

Follicular helper T ($T_{FH}$) cells are recently highlighted as their crucial role for humoral immunity to infection as well as their abnormal control to induce autoimmune disease. During an infection, na$\ddot{i}$ve T cells are differentiating into $T_{FH}$ cells which mediate memory B cells and long-lived plasma cells in germinal center (GC). $T_{FH}$ cells are characterized by their expression of master regulator, Bcl-6, and chemokine receptor, CXCR5, which are essential for the migration of T cells into the B cell follicle. Within the follicle, crosstalk occurs between B cells and $T_{FH}$ cells, leading to class switch recombination and affinity maturation. Various signaling molecules, including cytokines, surface molecules, and transcription factors are involved in $T_{FH}$ cell differentiation. IL-6 and IL-21 cytokine-mediated STAT signaling pathways, including STAT1 and STAT3, are crucial for inducing Bcl-6 expression and $T_{FH}$ cell differentiation. $T_{FH}$ cells express important surface molecules such as ICOS, PD-1, IL-21, BTLA, SAP and CD40L for mediating the interaction between T and B cells. Recently, two types of microRNA (miRNA) were found to be involved in the regulation of $T_{FH}$ cells. The miR-17-92 cluster induces Bcl-6 and $T_{FH}$ cell differentiation, whereas miR-10a negatively regulates Bcl-6 expression in T cells. In addition, follicular regulatory T ($T_{FR}$) cells are studied as thymus-derived $CXCR5^+PD-1^+Foxp3^+\;T_{reg}$ cells that play a significant role in limiting the GC response. Regulation of $T_{FH}$ cell differentiation and the GC reaction via miRNA and $T_{FR}$ cells could be important regulatory mechanisms for maintaining immune tolerance and preventing autoimmune diseases such as systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA). Here, we review recent studies on the various factors that affect $T_{FH}$ cell differentiation, and the role of $T_{FH}$ cells in autoimmune diseases.

• Asian-Australasian Journal of Animal Sciences
• /
• v.28 no.12
• /
• pp.1729-1735
• /
• 2015

This study was conducted to evaluate the effect of medicinal plant by-products (MPB) supplementation to a total mixed ration (TMR) on growth, carcass characteristics and economic efficacy in the late fattening period of Hanwoo steers. Twenty seven steers (body weight [BW], $573{\pm}57kg$) were assigned to 3 treatment groups so that each treatment based on BW contained 9 animals. All groups received ad libitum TMR throughout the feeding trial until slaughter (from 24 to 30 months of age) and treatments were as follows: control, 1,000 g/kg TMR; treatment 1 (T1), 970 g/kg TMR and 30 g/kg MPB; treatment 2 (T2), 950 g/kg TMR and 50 g/kg MPB. Initial and final BW were not different among treatments. Resultant data were analyzed using general linear models of SAS. Average daily gain and feed efficiency were higher (p<0.05) for T1 than control, but there was no difference between control and T2. Plasma albumin showed low-, intermediate- and high-level (p<0.05) for control, T1 and T2, whereas non-esterified fatty acid was high-, intermediate- and high-level (p<0.05) for control, T1 and T2, respectively. Carcass weight, carcass rate, backfat thickness and rib eye muscle area were not affected by MPB supplementation, whereas quality and yield grades were highest (p<0.05) for T1 and T2, respectively. Daily feed costs were decreased by 0.5% and 0.8% and carcass prices were increased by 18.1% and 7.6% for T1 and T2 compared to control, resulting from substituting TMR with 30 and 50 g/kg MPB, respectively. In conclusion, the substituting TMR by 30 g/kg MPB may be a potential feed supplement approach to improve economic efficacy in the late fattening period of Hanwoo steers.

• Applied Microscopy
• /
• v.32 no.4
• /
• pp.377-383
• /
• 2002

We have detected the murine zinc transporter, ZnT3, and zinc ions in the mouse choroid plexus by immunocytochemistry (ICC) and zinc selenium autometallography ($ZnSe^{AMG}$), respectively. BALB/c mice served as experimental animals. Routine floating ABC immunocytochemical procedures were used for the ZnT3 immunocytochemistry, and the mice were injected intraperitoneally (i.p.) with sodium selenide (10 mg/kg) for the zinc selenium autometallography. The choroid plexus showed weak immunoreactivity (Ir) for ZnT3. At high magnification, ZnT3-Ir was seen to be located in the choroid epithelium and the connective tissue of the capillaries. At the EM level, a high electron density of ZnT3-immunoreactivity was restricted to vesicle membranes as well as microvilli in the apical membrane. In contrast, immunostaining of ZnT3 was completely absent in the basolateral plasma membrane and other cell organelles. After silver enhancement, fine $ZnSe^{AMG}$ grains were observed in both the epithelial and endothelial cells of the choroid plexus. Few $ZnSe^{AMG}$ grains present in the cell bodies of the choroid epithelial cells were located in multivesicular bodies. It is striking that very many $ZnSe^{AMG}$ grains were observed in the endothelial cells of the capillaries. These findings establish the choroid plexus as a non-neuronal pool of zinc ions in the brain, although the functional significance of this pool is not clear. The choroid epithelium, however, may play an important role in the transportation of zinc between the CSF and brain tissue.

• Journal of Life Science
• /
• v.26 no.11
• /
• pp.1345-1354
• /
• 2016

Alopecia areata (AA) is a common and tissue-specific autoimmune disease of hair follicle resulting in the loss of hair on the scalp and elsewhere on the body. Hair follicles is a unique organ because it has its own immune system and hormonal milieu and has a different immune state at each hair cycle stage. The collapses of anagen-dependent hair follicle immune privilege arise autoimmune attack, inducing ectopic MHC class I expression in the hair follicle epithelium and autoantigen presentation to autoreactive CD8+T cells, which results in AA. Clinical and experimental studies have pointed that psychological stress may also influence the hair follicle immune/hormone systems and contribute to the induction of AA. The key pathogenesis of AA is associated with immune privilege guardians (including ACTH, ${\alpha}-MSH$, and $TGF-{\beta}$), natural killer group 2D-positive (NKG2D+) cells (including NK and CD8+T cells), and stress hormones (including CRH and substance P). Effective treatments for AA are still demanded. One of the future targets of treatment will be the modification of hair follicle immune privilege including stress. Recent studies have reported that JAK inhibitors and immunomodulators used in other autoimmune disease, such as psoriasis, atopic dermatitis, and rheumatoid arthritis, Tregs, platelet-rich plasma therapy, statins, and prostaglandin anaolgues are effective for AA. Here the article reviews the recent understanding in the pathogenesis associated with perifollicular endocrine/immunology and new treatments of AA.

• Parasites, Hosts and Diseases
• /
• v.56 no.1
• /
• pp.11-19
• /
• 2018

In Brazil, visceral leishmaniasis (VL) is expanding and becoming urbanized, especially in non-endemic areas such as the State of Rio Grande do Sul. Considering that infected dogs are the main reservoir for zoonotic VL, this study evaluated the prevalence of canine visceral leishmaniasis (CVL) in the metropolitan area of Porto Alegre, a new area of expansion of VL in Brazil. Serum and plasma from 405 asymptomatic dogs from the municipalities of Canoas (n=107), $S\tilde{a}o$ Leopoldo (n=216), and Novo Hamburgo (n=82) were tested for CVL using immunochromatographic ($DPP^{(R)}$) and ELISA $EIE^{(R)}$ assays (2 assays officially adopted by the Brazilian government for the diagnosis of CVL) and real-time PCR to confirm the results. There was no agreement among serological and real-time PCR results, indicating that the Leishmania infection in asymptomatic animals with low parasite load, confirmed by negative parasitological tests (smears and parasite culture), need to be evaluated by molecular methods. The prevalence of LVC in the metropolitan region of Porto Alegre, confirmed by real-time PCR was 4% (5.6% in Canoas and 4.6% in $S\tilde{a}o$ Leopoldo). The use of molecular method is essential for accurate diagnosis of CVL, especially in asymptomatic dogs in non-endemic areas.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.26 no.2
• /
• pp.308-313
• /
• 1997

To investigate the effects of Angelica keiskei Koidz on cholesterol metabolism, male Sprague Dawley rats were fed diets containing 5% of A. keiskei flour for 6 weeks. A. keiskei decreased plasma contents of total cholesterol, LDL-cholesterol and triglyceride while showing no change in HDL-cholesterol, resulting in reduction of atherosclerotic index. Decrease in liver concentration of triglyceride and increases in fecal excretion of cholesterol, total neutral steroid and bile acid were also observed. Those results suggest that A. keiskei improves hypercholesterolemia through, at least in part, reducing the absorption of cholesterol and enhancing the catabolism of cholesterol to bile acids.