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Role of murine Peyer's patch lymphocytes against primary and challenge infections with Cryptosporidium parvum

  • Guk, Sang-Mee;Chai, Jong-Yil
    • Parasites, Hosts and Diseases
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    • v.45 no.3
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    • pp.175-180
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    • 2007
  • In order to determine the role of Peyer's patch lymphocytes (PPL) in self-clearing of Cryptosporidium parvum infection in murine models, changes in PPL subsets, their cytokine expression, and in vitro IgG1 and IgA secretions by PPL were observed in primary- and challenge-infected C57BL/6 mice. In primary-infected mice, the percentages of CD4+ T cells, CD8+ T cells, slgA+ B cells, IL-2+ T cells, and $IFN-{\gamma}+$ T cells among the PPL, increased significantly (P < 0.05) on day 10 post-infection (PI). Secretion of IgG1 and IgA in vitro by PPL also increased on day 10 PI. However, all these responses, with the exception of IgG1 and IgA secretions, decreased in challenge-infected mice on day 7 post-challenge (= day 13 PI); their IgG1 and IgA levels were higher (P > 0.05) than those in primary-infected mice. The results suggest that murine PPL play an important role in self-clearing of primary C. parvum infections through proliferation of CD4+, CD8+, IL-2+, and $IFN-{\gamma}+$ T cells, and IgG1 and IgA-secreting 8 cells. In challenge infections, the role of T cells is reduced whereas that of 8 cells secreting IgA appeared to be continuously important.

The effect of human mesenchymal stem cell injection on pain behavior in chronic post-ischemia pain mice

  • Yoo, Sie Hyeon;Lee, Sung Hyun;Lee, Seunghwan;Park, Jae Hong;Lee, Seunghyeon;Jin, Heecheol;Park, Hue Jung
    • The Korean Journal of Pain
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    • v.33 no.1
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    • pp.23-29
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    • 2020
  • Background: Neuropathic pain (NP) is considered a clinically incurable condition despite various treatment options due to its diverse causes and complicated disease mechanisms. Since the early 2000s, multipotent human mesenchymal stem cells (hMSCs) have been used in the treatment of NP in animal models. However, the effects of hMSC injections have not been studied in chronic post-ischemia pain (CPIP) mice models. Here, we investigated whether intrathecal (IT) and intrapaw (IP) injections of hMSCs can reduce mechanical allodynia in CPIP model mice. Methods: Seventeen CPIP C57/BL6 mice were selected and randomized into four groups: IT sham (n = 4), IT stem (n = 5), IP sham (n = 4), and IP stem (n = 4). Mice in the IT sham and IT stem groups received an injection of 5 μL saline and 2 × 104 hMSCs, respectively, while mice in the IP sham and IP stem groups received an injection of 5 μL saline and 2 × 105 hMSCs, respectively. Mechanical allodynia was assessed using von Frey filaments from pre-injection to 30 days post-injection. Glial fibrillary acidic protein (GFAP) expression in the spinal cord and dorsal root ganglia were also evaluated. Results: IT and IP injections of hMSCs improved mechanical allodynia. GFAP expression was decreased on day 25 post-injection compared with the sham group. Injections of hMSCs improved allodynia and GFAP expression was decreased compared with the sham group. Conclusions: These results suggested that hMSCs may be also another treatment modality in NP model by ischemia-reperfusion.

Description of a Postflexion Larva Specimen of Liopropoma japonicum (Döderlein) off Jeju Island, Korea (제주도 근해에서 채집된 가시우럭(Liopropoma japonicum) 후기자어의 외부형태 관찰)

  • Kim, Sung;Yoo, Jae-Myung;Lee, Eun-Kyung
    • Ocean and Polar Research
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    • v.26 no.4
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    • pp.685-688
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    • 2004
  • The morphology of the post-larva of Liopropoma japonicum of the family serranidae is described from one specimen collected off Jeju Island ($32^{\circ}42'N,\;127^{\circ}45'E$) on May 3, 2001. The body is laterally compressed and caudal peduncle deep at 19.4 mm BL. Anus locates at a vertical through posteriormost(8th) dorsal-fin spine. Dorsal fin rays were VIII, 14; anal fin rays III, 10; pectroal fin rays 16; pelvic fin rays I, 5. A caudal fin is rounded. Length of elongate second and third dorsal-fin spines are 499% and 423% of the body length, respectively. A series of 50 sensory pores on body is elongated to caudal fin. Head spination is well developed. Melanophores are moderately large and dense in the head, and are small and rare in teh neck. Being freshly collected, the body color is yellowish-red and tail is semitransparent. The head is yellowish-red and the brain is dark-red.

Influence of Aspirin on Pilocarpine-Induced Epilepsy in Mice

  • Jeong, Kyoung Hoon;Kim, Joo Youn;Choi, Yun-Sik;Lee, Mun-Yong;Kim, Seong Yun
    • The Korean Journal of Physiology and Pharmacology
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    • v.17 no.1
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    • pp.15-21
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    • 2013
  • Aspirin (acetylsalicylic acid) is one of the most widely used therapeutic agents based on its pharmacological actions, including anti-inflammatory, analgesic, anti-pyretic, and anti-thrombotic effects. In this study, we investigated the effects of aspirin on seizure susceptibility and hippocampal neuropathology following pilocarpine-induced status epilepticus (SE). SE was induced by pilocarpine hydrochloride (280 mg/kg, i.p.) administration in C57BL/6 mice (aged 8 weeks). Aspirin was administered daily (15 mg/kg or 150 mg/kg, i.p.) for 10 days starting 3 days before SE, continuing until 6 days after SE. After pilocarpine injection, SE onset time and mortality were recorded. Neuronal cell death was examined using cresyl violet and Fluoro-Jade staining, and glial responses were observed 7 days post SE using immunohistochemistry. In the aspirin-treated group, the onset time of SE was significantly shortened and mortality was markedly increased compared to the control group. However, in this study, aspirin treatment did not affect SE-induced neuronal cell death or astroglial and microglial responses in the hippocampus. In conclusion, these results suggest that the safety of aspirin should be reevaluated in some patients, especially with neurological disorders such as temporal lobe epilepsy.

Expression and Characterization of Recombinant E2 Protein of Hepatitis C Virus by Insect Cell/Baculovirus Expression System

  • Han, Bong-Kwan;Lee, Bum-Yong;Min, Mi-Kyung;Jung, Kyung-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.8 no.4
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    • pp.361-368
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    • 1998
  • The E2 protein of HCV (hepatitis C virus) is thought to have a potential role in the development of subunit vaccines and diagnostics. To express it by the insect cell/baculovirus expression (Bacu) system, we constructed a recombinant Autographa californica nuclear polyhedrosis virus (AcIL3E2), determined the most appropriate expression conditions in terms of host cell line and culture medium, and characterized the expressed HCV E2 protein. A culture system using Trichoplusia ni BTI-TN5Bl-4 cells and SF 900IISFM medium expressed a relatively high level of HCV E2 protein. It was revealed that its glycosylation properties and subcellular localization were almost the same as the ones in the mammalian cell expression system previously reported, suggesting the recombinant HCV E2 protein derived from our Bacu system can be utilized for development of a subunit vaccine and diagnostics. Interestingly, HCV E2 protein was not degraded at all even at 43 h post-heat shock in the heat shock-induced necrotic cells, probably due to its integration into the microsomal membrane, indicating that heat shock can be employed to purify HCV E2 protein.

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Time gap between oocyst shedding and antibody responses in mice infected with Cryptosporidium parvum

  • Yu, Jae-Ran;Lee, Soo-Ung
    • Parasites, Hosts and Diseases
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    • v.45 no.3
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    • pp.225-228
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    • 2007
  • We observed the time gap between oocyst shedding and antibody responses in mice (3-week-old C57BL/6J females) infected with Cryptosporidium parvum. Oocyst shedding was verified by modified acid-fast staining. The individually collected mouse sera were assessed for C. parvum IgM and IgG antibodies by enzyme-linked immunosorbent assay from 5 to 25 weeks after infection. The results showed that C. parvum oocysts were shed from day 5 to 51 post-infection (PI). The IgM antibody titers to C. parvum peaked at week 5 PI, whereas the IgG antibody titers achieved maximum levels at week 25 PI. The results revealed that IgM responses to C. parvum infection occurred during the early stage of infection and overlapped with the oocyst shedding period, whereas IgG responses occurred during the late stage and was not correlated with oocyst shedding. Hence, IgM antibody detection may prove helpful for the diagnosis of acute cryptosporidiosis, and IgG antibody detection may prove effective for the detection of past infection and endemicity.

Extracellular Overproduction of $\beta$-Cyclodextrin Glucanotransferase in a Recombinant E. coli Using Secretive Expression System

  • Lee, Kwang-Woo;Shin, Hyun-Dong;Lee, Yong-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.12 no.5
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    • pp.753-759
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    • 2002
  • $\beta$-Cyclodextrin glucanotransferase ($\beta$-CGTase) was overproduced extracellularly using recombinant E. coli by transforming the plasmid pECGT harboring a secretive signal peptide. The $\beta$-CGTase gene of alkalophilic Bacillus firmus var alkalophilus was inserted into the high expression vector pET20b(+) containing a secretive pelB signal peptide, and then transformed into E. coli BL2l(DE3)pLysS. The optimum culture conditions fer the overproduction of $\beta$-CGTase were determined to be TB medium containing 0.5% (w/v) soluble starch at post-induction temperature of $25^{\circ}C$. A significant amount of $\beta$-CGTase, up to 5.83 U/ml, which was nine times higher than that in the parent strain B. firmus var. alkalophilus, was overproduced in the extracellular compartment. A pH-stat fed-batch cultivation of the recombinant E. coli was also performed to achieve the secretive overproduction of $\beta$-CGTase at a high cell density, resulting in production of up to 21.6 U/ml of $\beta$-CGTase.

Characterization of the Open Reading Frame 35 of Bombyx mori Nucleopolyhedrovirus

  • Zhu, Ying Min;Li, Guo Hui;Yao, Qin;Chen, Ke Ping;Guo, Zhong Jian
    • International Journal of Industrial Entomology and Biomaterials
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    • v.21 no.2
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    • pp.157-162
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    • 2010
  • Open reading frame 35 (bm35) of the Bombyx mori nucleopolyhedrovirus (BmNPV) is a special gene whose homologues are only found in some group-I nucleopolyhedroviruses, suggesting that bm35 plays a specific role in the viral life cycle. This paper described the characterization of BmNPV bm35. Computerassisted sequence analysis shows that a putative RING finger motif is observed in the protein, Bm35 encoded by bm35. The coding sequence of bm35 was amplified and subcloned into the vector pET30a(+) and the $(His)_6$-tagged fusion protein His-Bm35 was expressed in the Escherichia coli BL21 (DE3) LysS cells. The bm35 transcript and Bm35 protein were detected in BmNPV-infected BmN cells at 12~48 h post infection (p.i.) by RT-PCR and Western blot analysis using the polyclonal antibody generated by immunizing a rabbit with purified $(His)_6$-tagged Bm35, suggesting that bm35 is synthesized in the late stage of BmNPV infection cycle. Bm35 was not a structural component associated with budded virus (BV) and occlusion derived virus (ODV). These data indicated that bm35 is a functional gene in the BmNPV life cycle.

In vitro and in vivo inhibition of Helicobacter pylori by Lactobacilllus paracasei HP7

  • Hong, Seong-Soo;Lee, Hyun-A;Kim, Joo Yun;Jeong, Ji-Woong;Shim, Jae-Jung;Lee, Jung Lyoul;Sim, Jae-Hun;Chung, Yungho;Kim, Okjin
    • Laboraroty Animal Research
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    • v.34 no.4
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    • pp.216-222
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    • 2018
  • The efficacy of standard therapeutic strategies for Helicobacter pylori (H. pylori) infection is decreasing over time due to the emergence of drug-resistant strains. As an alternative, the present study investigated the capacity of Lactobacilllus paracasei (L. paracasei) HP7, isolated from kimchi, to inhibit H. pylori growth. The effects of L. paracasei HP7 on H. pylori adhesion and H. pylori-induced inflammation were examined in AGS human gastric adenocarcinoma epithelial cells and a mouse model of H. pylori SS1 infection. L. paracasei HP7 reduced H. pylori adhesion to AGS cells and suppressed the inflammatory response in infected cells by downregulating interleukin-8. H. pylori colonization in the stomach of C57BL/6 mice was demonstrated by rapid urease test, and results showed significant decrease in mice post-treated with L. paracasei HP7. Additionally, L. paracasei HP7 decreased gastric inflammation and epithelial lesions in the stomach of H. pylori-infected mice. These results demonstrate that L. paracasei HP7 treatment can inhibit H. pylori growth and is thus a promising treatment for patients with gastric symptoms such as gastritis that are caused by H. pylori infection.

Phosphodiesterase III Inhibitor Cilostazol Protects Amyloid β-Induced Neuronal Cell Injury via Peroxisome Proliferator-Activated Receptor-γ Activation (Amyloid β에 의해 유도된 신경세포 손상에 대한 phosphodiesterase III inhibitor인 cilostazol의 신경보호 효과)

  • Park, Sun-Haeng;Kim, Ji-Hyun;Bae, Sun-Sik;Hong, Ki-Whan;Choi, Byung-Tae;Shin, Hwa-Kyoung
    • Journal of Life Science
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    • v.21 no.5
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    • pp.647-655
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    • 2011
  • The neurotoxicity of aggregated amyloid ${\beta}$ ($A{\beta}$) has been implicated as a critical cause in the pathogenesis of Alzheimer's disease (AD). It can cause neurotoxicity in AD by evoking a cascade of apoptosis to neuron. Here, we investigated the neuroprotective effects of cilostazol, which acts as a phosphodiesterase III inhibitor, on $A{\beta}_{25-35}$-induced cytotoxicity in mouse neuronal cells and cognitive decline in the C57BL/6J AD mouse model via peroxisome proliferator-activated receptor (PPAR)-${\gamma}$ activation. $A{\beta}_{25-35}$ significantly reduced cell viability and increased the number of apoptotic-like cells. Cilostazol treatment recovered cells from $A{\beta}$-induced cell death as well as rosiglitazone, a PPAR-${\gamma}$ activator. These effects were suppressed by GW9662, an antagonist of PPAR-${\gamma}$ activity, indicative of a PPAR-${\gamma}$-mediated signaling. In addition, cilostazol and rosiglitazone also restored PPAR-${\gamma}$ activity levels that had been altered as a result of $A{\beta}_{25-35}$ treatment, which were antagonized by GW9662. Furthermore, cilostazol also markedly decreased the number of apoptotic-like cells and decreased the Bax/Bcl-2 ratio. Intracerebroventricular injection of $A{\beta}_{25-35}$ in C57BL/6J mice resulted in impaired cognitive function. Oral administration of cilostazol (20 mg/kg) for 2 weeks before $A{\beta}_{25-35}$ injection and once a day for 4 weeks post-surgery almost completely prevented the $A{\beta}_{25-35}$-induced cognitive deficits, as did rosiglitazone. Taken together, our findings suggest that cilostazol could attenuate $A{\beta}_{25-35}$-induced neuronal cell injury and apoptosis as well as promote the survival of neuronal cells, subsequently improving cognitive decline in AD, partly because of PPAR-${\gamma}$ activation. The phosphodiesterase III inhibitor cilostazol may be the basis of a novel strategy for the therapy of AD.