• 제목/요약/키워드: in vitro infection method

검색결과 77건 처리시간 0.021초

Expression on Echinostoma hortense Infection Manifestation on the Cytokine of the Splenocytes of BALB/c and C3H/HeN Mice

  • Ryang Yong-Suk;Cho Yoon-Kyung;IM Jee-Aee;Lee Kyu-Je;Kim Insik;Sung Ho-Joong
    • 대한의생명과학회지
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    • 제10권3호
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    • pp.219-230
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    • 2004
  • This experiment was performed to examine the in vitro and in vivo affects of the two different haplotype strains of mice, BALB/c and C3H/HeN infected with Echinostoma hortense, and the manifestation of the profiles of cytokine in the splenocytes. In the in vitro experiment, the two mice's splenocytes were divided and stimulated with antigen of crude extracts and the antigen of excretory and secretory products of an adult warm and the manifestation of cytokine mRNA was verified with RT-PCR. As a result, the two different strains of mice both strongly manifested the Th2 cytokine rather than the Thl cytokine and in the case of the Th2 cytokine, the BALB/c mice manifested more strongly than the C3H/HeN mice. In the experiment using the ELISA method, the protem cytokine manifestation had the same result as the mRNA experiment. In the in vivo experiment, the mice was infected via oral route with the metacercaria of the Echinostoma hortense and the manifestation of cytokine was verified by RT-PCR and ELISA and the results were the same as the in vitro experiment. Therefore, in the two strains of BALB/c and C3H/HeN, the C3H/HeN showed a higher susceptivity to the Echinostoma hortense.

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Anti-Toxoplasma Activities of Zea Mays and Eryngium Caucasicum Extracts, In Vitro and In Vivo

  • Ahmadpour, Ehsan;Ebrahimzadeh, Mohammad Ali;Sharif, Mehdi;Edalatian, Sara;Sarvi, Shahabeddin;Montazeri, Mahbobeh;Mehrzadi, Saeed;Akbari, Mohammad;Rahimi, Mohammad Taghi;Daryani, Ahmad
    • 대한약침학회지
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    • 제22권3호
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    • pp.154-159
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    • 2019
  • Objectives: Toxoplasmosis is a worldwide health problem that caused by intracellular apicomplexan parasite, Toxoplasma gondii (T. gondii). Considering that the available drugs for toxoplasmosis have serious host toxicity, the aim of the current study was to survey the in vitro and in vivo anti-Toxoplasma activity of Zea mays (Z. mays) and Eryngium caucasicum (E. caucasicum) extracts. Methods: Four concentrations (5, 10, 25, and $50mg\;mL^{-1}$) of Z. mays and E. caucasicum methanolic extracts for 30, 60, 120, and 180 min were incubated with infected macrophages and then the viability of RH strain of T. gondii tachyzoites was evaluated by trypan blue staining method. Also, we evaluated the survival rate of acutely infected mice with the extracts (100 and $200mg\;kg^{-1}\;day^{-1}$) intraperitoneally for 5 days after infection with $2{\times}104$ tachyzoites of T. gondii. Results: The anti-Toxoplasma effect of the methanolic extracts were extremely significant compared to the negative control group in all exposure times (P < 0.05). The Z. mays (10, 25 and $50mg\;mL^{-1}$) killed 100% of the parasites after 180 and 120 min exposure, respectively. Also, high toxoplasmacidal activity was observed with E. caucasicum extract. Furthermore, treatment of experimentally infected mice with the Z. mays (100, $200mg\;kg^{-1}\;day^{-1}$) and E. caucasicum ($100mg\;kg^{-1}\;day^{-1}$) significantly increased their survival rate compared to untreated infected control (P < 0.05). Conclusion: These extracts are promising candidates for further medicine development on toxoplasmosis. However, further investigations are necessary to clarify effective fractions of the Z. mays and E. caucasicum extracts and the mechanisms of action.

In Vitro and in Vivo Wound Healing Properties of Plasma and Serum from Crocodylus siamensis Blood

  • Jangpromma, Nisachon;Preecharram, Sutthidech;Srilert, Thanawan;Maijaroen, Surachai;Mahakunakorn, Pramote;Nualkaew, Natsajee;Daduang, Sakda;Klaynongsruang, Sompong
    • Journal of Microbiology and Biotechnology
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    • 제26권6호
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    • pp.1140-1147
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    • 2016
  • The plasma and serum of Crocodylus siamensis have previously been reported to exhibit potent antimicrobial, antioxidant, and anti-inflammatory activities. During wound healing, these biological properties play a crucial role for supporting the formation of new tissue around the injured skin in the recovery process. Thus, this study aimed to evaluate the wound healing properties of C. siamensis plasma and serum. The collected data demonstrate that crocodile plasma and serum were able to activate in vitro proliferation and migration of HaCaT, a human keratinocyte cell line, which represents an essential phase in the wound healing process. With respect to investigating cell migration, a scratch wound experiment was performed which revealed the ability of plasma and serum to decrease the gap of wounds in a dose-dependent manner. Consistent with the in vitro results, remarkably enhanced wound repair was also observed in a mouse excisional skin wound model after treatment with plasma or serum. The effects of C. siamensis plasma and serum on wound healing were further elucidated by treating wound infections by Staphylococcus aureus ATCC 25923 on mice skin coupled with a histological method. The results indicate that crocodile plasma and serum promote the prevention of wound infection and boost the re-epithelialization necessary for the formation of new skin. Therefore, this work represents the first study to demonstrate the efficiency of C. siamensis plasma and serum with respect to their wound healing properties and strongly supports the utilization of C. siamensis plasma and serum as therapeutic products for injured skin treatment.

Ultraviolet-C 조사의 살균 효과 (The Effect of Ultraviolet-C Radiation on Disinfection)

  • 최홍식;최규환;박소연
    • 한국전문물리치료학회지
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    • 제9권3호
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    • pp.93-99
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    • 2002
  • Traditionally, ultraviolet (UV) has been used for treating the pressure sore and skin wound. The effects of UVA and UVB radiation on disinfection have been reported. The purpose of this study was to examine the effectiveness of UVC radiation on disinfection of Escherichia coli, Staphylococcus aureus, Salmonella typhimurium in vitro. Three bacterium were radiated by UVC (250 nm, 20 seconds) and incubated at $37^{\circ}C$ for 24 hours at the agar culture medium. Kill rates of all three bacterium were 99.9%. UVC radiated on three kinds of bacterium for 30 or 60 seconds. Kill rates were 99.9% both 30 and 60 seconds. This data suggests that UV light at 250 nm could be a useful method to minimize infection and shorten healing time in pressure sore and skin wound condition.

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무병종서 생산을 위한 감자X바이러스 및 엽권바이러스에 관한 연구 (Studies on the Potato Virus X and Potato Leaf Roll Virus for Disease-free Seed Potato Production)

  • 최정일
    • 한국작물학회지
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    • 제7권1호
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    • pp.31-63
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    • 1969
  • A series of experiment was carried out to study on the production of disease-free seed potatoes at the Alpine Experiment Station from 1960 to 1968, which initiated a study of comparison on degeneration of plain warm region and high altitude products and the effect of latent potato virus X (PVX) and potato leaf roll virus(PLRV) on degeneration. Particular observations were made on some aspect of the nature of potato virus disease and its control such as concentrations of PVX, range of host plants, physical properties such as concentrations of PYX, range of host plants, physical properties and carrying effect of insects, by investigating 9 different areas of the main potato producing regions (Kimhae, Taegu, Choongju, Taejoen, Suwon, Kwangju, Chonju, Cheju and Chinju). Highly purified anti-serum was separated and tested for control of the virus disease and also various method of prevention and control of PLRV were observed, using cultivation of sprouted seed tubers, early harvesting method, and systemic chemicals. The results obtained are summarized as follows; 1. Potato yield in the plain region decreased by 32.8~66.3% in the first year cultivation of seed potatoes from colder region, and the rate of virus infection was 92.9 to 95.4%. 2. Plants of three families including, 20 species were susceptible to the PVX, and among the plants Salvia officinalis of a habits only was the carrier while the symptom of Digitalis purpurea of Screphulariaceae was masked. Necrosis and ring spot was occurred in most pJants of the Solanaceae and ring spot symptom also was observed in Nicotiana tabacum L. var. White Burley and in N. glutinosa. 3. The 8$C_2$ strain of virus had the following physical properties; thermal inactivation point, 68-$72^{\circ}C$ : dilution inactivation point, above 1, 000, 000 dilution: ageing in vitro, 240-360 days: and ageing in dry plant tissue, 30 days. 4. Myzus persicae and Oxya spp. did not transmit the 8$C_2$ strain of potato virus. 5. Virus was purified through the ammonium sulphate isolating method, and higher titer value, 1/2048 was obtained through anti-serum test. 6. Inhibition Chenopodiacae on the virus infection of potato was remarkable, and inhibition of local lesion host also was observed. 7, By earlier planting of sprouted seed tubers, growth period could be prolonged by 10 to 12 days. 8. Earlier harvest decreased much the rate of virus infection of seed potatoes. 9. According to the results of aphid control trial using systemic soil insecticides at Kangnung and Taekwanlyung, PSP 204, Disyston and Thimet was effective to aphid control. In particular, control effect of twice treatments of PSP 204 was great. 10. Treatmental effect of those chemicals lasted about 60-70 days. However, single foliar application of emulsified chemicals was not effective to potato virus control. 11. The effect of PSP 204, Disyston, and Thimet on the control of potato leaf roll virus was great, particularly in the case of two treatments of PSP 204, at Kangnung as well as at Taekwanlyung. Higher negative correlationship between the control effect of potato leaf roll virus and potato yield was observed showing the value r=-0.85 at Kangnung, and r=-0.87 at Taekwanlyung. 12. Differences in the control effects among PSP 204, Disyston, and Thimet was not noticed.

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Development of a novel reverse transcription PCR and its application to field sample testing for feline calicivirus prevalence in healthy stray cats in Korea

  • Kim, Sung Jae;Park, Yong Ho;Park, Kun Taek
    • Journal of Veterinary Science
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    • 제21권5호
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    • pp.71.1-71.10
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    • 2020
  • Background: Feline calicivirus (FCV) is a major and highly infectious pathogen in cats worldwide. However, there have been limited studies about the status of FCV infections in Korea. Objectives: To investigate the current status of FCV infections in stray cats in Korea. Methods: A novel reverse transcription polymerase chain reaction (RT-PCR) assay was developed based on the conserved nucleotide sequences of reported FCV strains. Field swab samples were collected from 122 cats (2 hospital admitted cats and 120 stray cats) in 2016 and 2017. All the samples were tested by virus isolation and 2 different RT-PCRs, including the novel RT-PCR, for the detection of FCV. Results: The novel RT-PCR assay showed no cross-reactivity to the nucleic acids of the other feline pathogens tested, and the limit of detection was calculated as 100 TCID50/mL based on an in vitro assessment. The novel RT-PCR assay detected 5 positive samples from the 122 field samples, which showed perfect agreement with the results of the virus isolation method. In contrast, another RT-PCR assay used in a previous study in Korea detected no positive samples. The prevalence of FCV infection in stray cats was 2.5% (3/120) based on the results of virus isolation and the novel RT-PCR assays. Conclusions: The current study is the first report of the detection and prevalence of FCV in stray cats in Korea. The novel RT-PCR assay developed in this study showed high sensitivity and specificity, which indicates a useful diagnostic assay to identify FCV infection in cats.

A Rapid Radicle Assay for Prescreening Antagonistic Bacteria Against Phytophthora capsici on Pepper

  • Chang, Sung-Hwan;Kwack, Min-Sun;Kim, Yun-Sung;Lee, Jung-Yeop;Kim, Ki-Deok
    • Mycobiology
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    • 제29권4호
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    • pp.218-223
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    • 2001
  • A rapid radicle assay for prescreening antagonistic bacteria to Phytophthora capsic4 causal agent of Phytophthora blight of pepper was developed. Sixty-four bacterial strains with in vitro antifungal activity selected out of 1,400 strains isolated from soils of Ansung, Chunan, Koyang, and Paju, Korea in 1998 were used for development of the bioassay. Uniformly germinated pepper seeds dipped in bacterial cells for 3 hours were placed near the edges of growing mycelia of P. capsici on water agar containing 0.02% glucose. Five-week-old pepper plants(cv. Nockwang) were inoculated to compare with results of the radicle assay developed in this study. For plant inoculation, pepper seeds were sown in potting mixtures incorporated with the bacterial strains, then transplanted into steam-sterilized soils 3 weeks later. Plants were hole-inoculated with zoospores of P. capsici 2 weeks after transplanting. Disease incidence and severity were determined in radicle and plant assessments, respectively. In radicle assay, six strains, GK-B15, GK-B25, OA-B26, OA-B36, PK-B09, and VK-B14 consistently showed the significant(P=0.05) disease reduction against radicle infection by the fungus, four of which also did in plant assessments. Strains OA-B36 and GK-B15 consistently reduced the fungal infection in both the radicle assay and the plant assessment. Therefore, prescreening strains using the radicle assay developed in this study followed by plant assay could reduce time and labor, and improved the possibility of selecting antagonistic bacteria for control of Phytophthora blight of peppers.

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Effect of Temperature on the Killing of Opisthorchis viverrini Eggs In Vitro

  • Boueroy, Parichart;Duenngai, Kunyarat;Eamudomkarn, Chatanun;Sripan, Panupan;Boonmars, Thidarut;Pumhirunroj, Benjamabhorn;Artchayasawat, Atchara;Songsri, Jiraporn;Chomphumee, Kanpicha;Rattanasuwan, Panaratana;Laummaunwai, Porntip;Khueangchiangkhwang, Sukhonthip;Boonjaraspinyo, Sirintip
    • Parasites, Hosts and Diseases
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    • 제57권1호
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    • pp.49-53
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    • 2019
  • Contaminated liver fluke egg in the environment has led to the high prevalence of human opisthorchiasis associated with cholangiocarcinoma in Southeast Asia. To find the effective lessening methods of Opisthorchis viverrini eggs in the contaminated environment, we investigated the temperature conditions for killing of these trematode eggs in vitro. Numerous O. viverrini eggs were obtained in the proximal part of uteri of adult worms from experimental hamsters. Mature eggs with miracidium were allocated by experimental groups (2 control: positive and negative and 4 treatment: 50, 60, 70, and $80^{\circ}C$) with 0.85% saline, and treated by the experimental plan. Eggs in each experimental groups were observed under the confocal microscope after stain with Propidium Iodide (PI) to evaluate the effect of temperatures. Eggs in 70 and $80^{\circ}C$ groups were all killed after over 10 min heated. Majority of eggs in $60^{\circ}C$ (10, 15, and 30 min heated), 70 and $80^{\circ}C$ (5 min heated) groups were inactivated. However in $50^{\circ}C$ group, below half of eggs were to be killed in all time lapse (10, 15 and 30 min). In order to prevent O. viverrini infection and cholangiocarcinoma, direct treatment of sewage by heating at 70 or $80^{\circ}C$ at least 10 min is essential. Therefore, treatment of O. viverrini eggs at a high temperature is a potential method for controlling egg contamination in sewage.

젖소의 건유기 유방염 치료에 있어서 생분해 cephalexin microspheres의 효과 (Effects of Biodegradable Cephalexin Microspheres in Dry Cow Mastitis Therapy)

  • Hwang, Cheol-Yang
    • 한국임상수의학회지
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    • 제19권2호
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    • pp.228-235
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    • 2002
  • 유방염은 젖소에 있어서 유량의 감소, 유질 저하로 인한 우유 폐기문제, 젖소의 도태, 치료비 및 노동력의 부담등을 초래하는 가장 경제적 손실이 큰 질환이다. 현재까지 비유기 최종 착유 후에 항생제를 주입하는 건유기 치료법이 유방염 관리에 가장 효과적이며 폭넓게 쓰이고 있는 방법으로 알려져 있다. 하지만, 건유기 치료는 건유기 주입 항생제 제품들이 건유기 초기에만 지속적인 활성을 지니기 때문에 건유기 말기와 분만 전기에 있어서의 신규 감염은 방어할 수 없다. 따라서 본 연구는 건유기 유방염 관리를 위해 PLGA를 이용한 서방형 생분해 cephalexin microsphere를 제조하여 이의 임상적 효능을 평가하고자 하였다. PLGA는 무독성의 조직 반응을 일으키지 않는 특성으로 인해 약물 방출 조절 체계의 일환으로 인정을 받아왔다. 본 연구에서 cephalexin microsphere는 표면에 특징적인 구멍을 가지고 있는 구형 모양으로 확인이 되었으며 약물 방출 시험에서 초기 과다 방출 이후로 21일간 약물의 방출이 점차 감소한 뒤 3주와 4주 사이에 2차 방출을 보이는 맥동성 방출 양상이 관찰되었다. 현장적용 시험에서는 cephalexin microsphere를 주입한 실험군에서 대조군에 비해 분만 후 신규 감염율 및 치료율, 평균 체세포 수의 변화의 측면에서 볼 때 유의성 있는 차이가 있음을 확인하였다. 따라서 생분해 microsphere를 이용한 건유기 치료법은 건유기 동안 신규 감염을 예방하고 기감염을 감소시킴으로써 종전의 건유기 치료를 좀 더 효과적으로 증진시킬 수 있을 것으로 사료된다.

패혈증 치료제 개발을 위한 황백이 포함된 생약혼합제제 ABHC의 항균 효능 (Antibacterial Activity of Herbal Complex ABHC for Development of Novel Therapeutic Agent Against Sepsis)

  • 이기만;이금선;김유리;박준우;부경준;임동술;강태진
    • 생약학회지
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    • 제50권3호
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    • pp.191-197
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    • 2019
  • Sepsis, an infectious disease, is a life-threatening condition that arises when the response to infection causes injury to tissues and organs. The purpose of this study was to demonstrate whether ABHC-1 and ABHC-2, two functional extracts from herbal complex, have an anti-bacterial effect against Escherchia coli in vivo, in vitro experimental model. ABHC-1 and ABHC-2 showed the antibacterial activity against the bacteria by paper disc method. The minimum inhibitory concentration (MIC) was measured using alamar blue reagent. The MIC was shown at $60{\mu}g/ml$ from ABHC-1 and $500{\mu}g/ml$ from ABHC-2 against E. coli. We next examined the effect of ABHCs on the production of inflammatory cytokine, such as tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$), which is related to the induction of inflammation, in RAW 264.7 cell. ABHC-1 and ABHC-2 increased $TNF-{\alpha}$ production of RAW 264.7 cell in a dose-dependent manner while two extract decreased $TNF-{\alpha}$ production in lipopolysaccharide (LPS)-stimulated RAW 264.7 cell in a dose-dependent manner. At a dose of $1{\times}10^8$ E. coli. i.p., non-treated mice were succumbed, while most of mice treated with ABHC-1 were survived. Therefore, our results suggest that ABHC-1 has anti-bacterial activity and can be a novel therapeutic agent against infectious diseases.