• Clinical and Experimental Reproductive Medicine
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• v.48 no.3
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• pp.229-235
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• 2021

Objective: In this study, the effects of Lactobacillus acidophilus on testosterone (TES), follicle-stimulating hormone (FSH), luteinizing hormone (LH), androgen-binding protein (ABP), factor-associated apoptosis (FAS), and total cholesterol (TC), as well as histopathological changes, were investigated in male rats fed a high-cholesterol diet. Methods: The study included three groups. The control (C) group was fed standard-diet for 8 weeks. The hypercholesterolemia (HC) group was fed a 2% cholesterol-diet for 8 weeks. The therapeutic group (HCL) was fed a 2% cholesterol-diet for 8 weeks and administered L. acidophilus for the last 4 weeks. FSH, TES, and FAS levels in testicular tissue were determined using an enzyme-linked immunosorbent assay (ELISA), while another sample was examined histopathologically. LH and ABP levels were determined using ELISA, and serum TC levels were assessed via an autoanalyzer. Results: In the HC group, the TC levels were significantly higher and the LH levels were lower (p<0.05) than in the C group. The ABP levels were lower (p>0.05). In the HCL group, the LH and ABP levels were higher (p>0.05) and the TC level significantly lower (p<0.05) than in the HC group. The TES and FSH levels were lower, and the FAS levels were higher, in the HC than in the C group (p<0.05). In the HCL group, levels of all three resembled control levels. Histologically, in the testicular tissue of the HC group, the cells in the tubular wall exhibited atrophy, vacuolization, and reduced wall structure integrity. However, in the HCL group, these deteriorations were largely reversed. Conclusion: Supplementary dietary administration of an L. acidophilus to hypercholesterolemic male rats positively impacted testicular tissue and male fertility hormone levels.

• Korean Journal of Veterinary Service
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• v.43 no.4
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• pp.211-216
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• 2020

Coxiella burnetii is the causative agent of Q fever which is a zoonosis occuring in both humans and animals worldwide. The purpose of this study was to investigate the seroprevalence of C. burnetii in Korean native goat in Gyeongnam province, Korea. A total of 1,365 goat blood samples from 273 farms in Gyeongnam province were collected between 2018 and 2019. Among them, 177 (13.0%) samples out of 71 (26.0%) farms were seropositive for C. burnetii by ELISA. Seroprevalence were 15.4% and 10.9% in 2018 and 2019, respectively. According to the region, seroprevalence in western, central, eastern, northern and southern areas of Gyeongnam province were 16.6%, 17.8%, 8.0%, 11.6% and 10.8%, respectively. Seroprevalence was increased with breeding scale (Head<10:7.0%, 10≤Head<50:8.7%, 50≤Head<100:13.6%, 100≤Head:28.8%). Seroprevalence according to the season showed highest in summer (18.9%) and lowest in winter (9.4%). These results indicated that C. burnetii infection is widespread among Korean native goats of Gyeongnam province in Korea and further study needs to prevent the circulation of other livestock with Korean native goat.

• Parasites, Hosts and Diseases
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• v.61 no.4
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• pp.418-427
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• 2023

Toxoplasma gondii infections are primarily diagnosed by serological assays, whereas molecular and fluorescence-based techniques are garnering attention for their high sensitivity in detecting these infections. Nevertheless, each detection method has its limitations. The toxoplasmosis detection capabilities of most of the currently available methods have not been evaluated under identical experimental conditions. This study aimed to assess the diagnostic potential of enzyme-linked immunosorbent assay (ELISA), real-time polymerase chain reaction (RT-PCR), immunohistochemistry (IHC), and immunofluorescence (IF) in BALB/c mice experimentally infected with various doses of T. gondii ME49. The detection of toxoplasmosis from sera and brain tissues was markedly enhanced in mice subjected to high infection doses (200 and 300 cysts) compared to those subjected to lower doses (10 and 50 cysts) for all the detection methods. Additionally, increased B1 gene expression levels and cyst sizes were observed in the brain tissues of the mice. Importantly, IHC, IF, and ELISA, but not RT-PCR, successfully detected T. gondii infections at the lowest infection dose (10 cysts) in the brain. These findings may prove beneficial while designing experimental methodologies for detecting T. gondii infections in mice.

• Parasites, Hosts and Diseases
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• v.47 no.3
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• pp.219-225
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• 2009

The seroprevalence of cryptosporidiosis was examined using patients' sera collected from hospitals located in 4 different areas of the Republic of Korea. ELISA was used to measure antibody titers against Cryptosporidium parvum antigens from a total of 2,394 serum samples, which were collected randomly from patients in local hospitals; 1)Chungbuk National University Hospital, 2) Konkuk University Hospital, 3) local hospitals in Chuncheon, Gangwon-do (province), 4) Jeonnam National University Hospital, from 2002 through 2003. Of the2,394 samples assayed, 34%, 26%, and 56% were positive for C. parvum-specific IgG, IgM, and IgA antibodies, respectively, Positive IgG titers were most common in sera from Jeonnam National University Hospital, Gwangju, Jeollanam-do, and positive IgM titers were most common in sera from Chungbuk National University Hospital, Cheongju, Chuncheonqbuk-do. The seropositivity was positively correlated with age for both the IgG and IgA antibodies but was negatively correlated with age for the IgM antibodies. Western blotting revealed that 92%, 83%, and 77% of sera positive for IgG, IgM, and IgA ELISA reacted with 27-kDa antigens, respectively. These results suggested that infection with Cryptosporidium in hospital patients occurs more commonly than previously reported in the Republic of Korea.

• BMB Reports
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• v.29 no.3
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• pp.192-199
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• 1996

A rapid and sensitive method has been developed to detect a-fetoprotein (AFP) in human serum by a two-site sandwich enzyme-linked immunosorbent assay (ELISA) with monoclonal antibodies (MAbs) for human AFP within 1 h. To obtain the most sensitive and reliable MAbs. 12 kinds of MAbs (HPJ1 to HPJ12) as a capture antibody and 4 kinds of horseradish peroxidase (HRP) conjugated MAbs as a tracer antibody were investigated. Among these, only HPJ 10-HRP conjugated HPJ 1 (HPJ 10-HPJ $1^*$) and HPJ 11-HRP conjugated HPJ 10 (HPJ 11-HPJ $10^*$) were chosen as candidates based on the linearity of the standard curve and the sensitivity of the assay. To further characterize these two pairs. MAbs against human AFP were purified from hybridoma cells. conjugated with HRP. and then characterized to optimize the two-site sandwich ELISA The HPJ 10-HPJ $1^*$ pair showed a sensitivity of 1 ng/ml and a better reproducibility than the HPJ 11-HPJ $10^*$ pair when the human sera were incubated at $37^{\circ}C$ for 30 min. The results obtained for 480 randomly selected human sera showed 0~20 ng/ml of AFP values for the normal human sera. To test the utility of our kit, AFP concentrations were determined for 951 human sera (including 85 normal sera, 480 random blood sera, 213 HBsAg-positives. 50 anti-HCV antibody positives. and 47 malignant diseases) and compared with other commercially available AFP detecting kits. These results show that the present two-site sandwich ELISA method is a rapid, sensitive, and reliable procedure for detecting AFP in human serum.

• Journal of Microbiology and Biotechnology
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• v.21 no.7
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• pp.719-727
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• 2011

Botulism is a neuroparalytic disease caused by Clostridium botulinum, which produces seven (A-G) antigenically diverse neurotoxins (BoNTs). BoNTs are the most poisonous substances known to humans, with a median lethal dose ($LD_{50}$) of approximately 1 ng/kg of body weight. Owing to their extreme potency and lethality, they have the potential to be used as a bioterrorism agent. The mouse bioassay is the gold standard for the detection of botulinum neurotoxins; however, it requires at least 3-4 days for completion. Attempts have been made to develop an ELISA-based detection system, which is potentially an easier and more rapid method of botulinum neurotoxin detection. The present study was designed using a synthetic gene approach. The synthetic gene encoding the catalytic domain of BoNT serotype B from amino acids 1-450 was constructed with PCR overlapping primers (BoNT/B LC), cloned in a pQE30 UA vector, and expressed in an E. coli M15 host system. Recombinant protein production was optimized at 0.5 mM IPTG final concentration, 4 h post induction, resulting in a maximum yield of recombinant proteins. The immunogenic nature of the recombinant BoNT/B LC protein was evaluated by ELISA. Antibodies were raised in BALB/c mice using various adjuvants. A significant rise in antibody titer (p<0.05) was observed in the Alum group, followed by the Titermax Classic group, Freund's adjuvant, and the Titermax Gold group. These developed high-titer antibodies may prove useful for the detection of botulinum neurotoxins in food and clinical samples.

• Ocean and Polar Research
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• v.41 no.2
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• pp.79-88
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• 2019

In this study, the inhibitory effect of Atriplex gmelinii C. A. Mey. against the activity of MMP-2 and MMP-9 secreted from phorbol-12-myristate-13-acetate (PMA)-stimulated HT-1080 cells was evaluated by gelatin zymography and enzyme-linked immunosorbent assay (ELISA), reverse transcription polymerase-chain reaction (RT-PCR), and Western blot assay. Specimens of the halophyte A. gmelinii were extracted twice for 24 hr with methylene chloride ($CH_2Cl_2$), and then twice with methanol (MeOH), in turn. Each extract significantly inhibited the enzymatic activities in gelatin zymography and MMP ELISA kit, and expression of MMP-2 and 9 in mRNA and protein levels. Two crude extracts were combined and then the combined crude extracts were fractionated into n-hexane, 85% aqueous methanol (85% aq.MeOH), n-butanol (n-BuOH), and water ($H_2O$) fractions, according to solvent polarity. Among solvent-partitioned fractions, the 85% aq.MeOH fraction showed the strongest inhibitory effect against MMP-2 and -9 in gelatin zymography and MMP ELISA kit. In RT-PCR, all solvent-partitioned fractions significantly suppressed mRNA expression of MMP-2 and -9. On the other hand, in Western blot assay, all solvent-partitioned fractions except $H_2O$ significantly reduced expression levels of protein. HT 1080 cell migration was most significantly inhibited by the n-BuOH fraction followed by the 85% aq.MeOH and $H_2O$ fractions. These results suggest that A. gmelinii could be used as a potential source to inhibit tumor cell metastasis.

• IMMUNE NETWORK
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• v.1 no.2
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• pp.170-178
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• 2001

Background: Korean mistletoe (Viscum album) extract has been found to posses immunostimulatory activity. In this study, Korean mistletoe extract, M11C (non-lectin components), was used to know whether this extract might activate mouse peritoneal macrophages to produce interleukin $1{\beta}$ (IL-$1{\beta}$). Methods: Hemagglutination assay was carried out to examine whether M11C contained a lectin or not. To know the effect of M11C on the production of IL-$1{\beta}$, the macrophages were treated by the M11C, and then collected the supernatant (M11C stimulated macrophages-conditioned media; MMCM). MMCM was analyzed for the IL-$1{\beta}$ quantification and mRNA expression by means of ELISA and RT-PCR, respectively. Results: Maximum effective dose and time of M11C on IL-$1{\beta}$ production from macrophages were $20{\mu}g/m{\ell}$ and 8 hours, respectively. This ELISA data was reconfirmed by immunoblotting assay. indicating that M11C is a good candidate for an immunomodulator. The dose and time dependent effects of M11C on the expression of IL-$1{\beta}$ mRNA from macrophages was also shown in expression of mRNA detected by RT-PCR. Treatment dose and time for the maximum expression of IL-$1{\beta}$ mRNA were $20{\mu}g/m{\ell}$ and 4 hours, respectively. Maximum gene expression of IL-$1{\beta}$ was much earlier than maximum production of it. Conclusion: As results, Korean mistletoe extract, M11C, may be used for an immunomodulator. This will be able to make up for and solve the problems caused by existent immunoagent with many adverse effects through many other studies in future including one molecule extraction.

• BMB Reports
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• v.40 no.4
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• pp.459-466
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• 2007

The cDNA sequence of the Japanese flounder (Paralychthys olivaceus) IgD has been previously reported (GenBank accession no. AB052658) and this was followed by the detection of IgD mRNA expression in some flounder organ tissues. However, it has not been determined whether the flounder IgD gene is virtually expressed into IgD protein. To characterize the flounder immunoglobulins utilized in elucidating the mechanism, evolution and diversity of the flounder immune system, antibodies specific to IgD and IgM were necessary. In the present study, partial flounder recombinant IgD (rIgD), IgM (rIgM) and the conserved regions of IgD and IgM (rCIg) were produced by cloning the cDNA sequence using isotype specific primers which were designed to produce unique fragments of IgD and IgM specific amino acid sequences. The production of recombinant Igs was ascertained by SDS-gel electrophoresis and immunoblot analysis using anti-T7$\cdot}$Taq antibody. The produced recombinant Igs were purified using affinity columns, and used as immunogens. Antibodies specific to the isotype of flounder Igs were generated by immunizing rabbits with rfIgs and the antibodies produced were identified by enzyme-linked immunosorbent assay (ELISA) and immunoblotting. Specificities of the generated antibodies were evaluated by testing cross-reactivity between recombinant IgM and IgD. By ELISA, rabbit antibodies against the rfIgD fragment (anti-rfIgD) failed to recognize any kind of flounder serum Igs, whereas respective antibodies against rfCIg (anti-rfCIg) and rfIgM fragments (anti-rfIgM) reacted with serum Igs. Likewise, in immunoblot assays, though anti-rfIgD did not, both anti-rfCIg and anti-rfIgM bound with the ~85 kd flounder IgM heavy chain. By flow cytometry analysis, anti-rfCIg, anti-rfIgD and anti-rfIgM reacted with 6%, 3% and 6.5% of cells, respectively, suggesting that flounder IgD is not secreted in serum but expressed on flounder B-like cell surfaces as in mammals. Antibodies produced against recombinant flounder Igs could be used to develop sandwich assay systems for detecting flounder Igs and for further investigating the flounder immune system.

• Journal of Food Hygiene and Safety
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• v.16 no.4
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• pp.274-279
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• 2001

Generally, non-aflatoxigenic fungi, such as Aspergillus oryzae, and Aspergillus are main microflora in Korean traditional fermented foods including Meju and soybean paste, but sometimes, Aspergillus flavus and Aspergillus parasiticus can be contaminated and accumulated aflatoxins during fermentation and storage. So the screening of aflatoxigenic strains in fermented traditional food is very important to improve the sanitary quality of those foods. In this work, we screened aflatoxin producing fungi from commercial Meju and soybean paste in Western Gyeongnam by immunoassay. Samples were randomly purchased from market of the commercial Meju(10 EA) and soybean paste(20 EA) in nine areas of Western Gyeongnam. Of the samples collected,24 strains and 22 strains of Aspergillus sp. were isolated from Meju and soybean paste, respectively. The isolated strains were cultured on SLS media at $25^{\circ}C$ for 15 days. The cultured broth were extracted with ethyl acetate and were analysed to determine aflatoxin B$_1$(AFB$_1$) by direct competitive ELISA(DC-ELISA). Six strains(25%) isolated from Meju, and 2 strains(9%) isolated from saybean paste, were confined as aflatoxin producing strains. The average range of aflatoxin productivity of isolates from Meju was 54.6 $\pm$ 38.7 ng/ml and that from soybean paste was 11.1 $\pm$ 8.6 ng/ml, respectively. Among them, isolated strain No. M-5-4 produced a high level of AFBl and showed 98.26 ng/ml of AFB$_1$. Every isolates were also re-confined their AFB$_1$productivity by thin layer chromatography(TLC). The TLC results also showed same trend as DC-ELISA results. As the above results, the screening of hazard mycotoxigenic fungi from traditional fermented foods should be necessary for the safety and the application of HACCP system in the food manufactory in Korea.