• Journal of Food Hygiene and Safety
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• v.38 no.6
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• pp.442-448
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• 2023

This study aimed to prepare kombucha, a fermented tea beverage, containing Dendropanax morbiferus (DM) leaves and roots, and analyze its antioxidant and intracellular activities. We compared the pH change, total acidity, radical scavenging activity, and oxygen radical absorbance capacity (ORAC) of kombucha fermented with black tea alone and that with added DM leaves or roots during fermentation. Using RAW 264.7, we evaluated the effects of kombucha containing different DM parts on nitric oxide (NO) production and inflammation-related cytokine content in cells. Kombucha containing ethanol extracts of DM leaves (BTK-E-DML) and roots (BTK-E-DMR) showed higher radical scavenging activity and ORAC 3 d after fermentation than that prepared from black tea alone (BTK-Ori). In an in vitro experiment using RAW 264.7, samples were treated with 8 mg/mL kombucha considering cytotoxicity; the lipopolysaccharide (LPS)-induced NO content significantly reduced after BTK-E-DML and BTK-EDMR treatments compared with that after BTK-Ori treatment. Additionally, the levels of interleukin-6 and tumor necrosis factor-alpha, which were LPS-stimulated inflammatory cytokines, significantly decreased in cells treated with BTK-E-DML and BTK-E-DMR 15 d after fermentation compared with those treated with BTK-Ori. In conclusion, these results demonstrate that kombucha fermented with the leaves and roots of DM increases antioxidant activity and can significantly regulate inflammatory responses at the cellular level.

• Microbiology and Biotechnology Letters
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• v.11 no.3
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• pp.223-231
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• 1983

The productions of beta-exotoxin from sixteen Bacillus thuringiensis strains were examined by Micrococus flava primarily, and then measured by spectrophotometer during culturing in Conner and Hansen mineral salts medium at 28$^{\circ}C$. Also the toxic effects of the toxin to mice were checked. The growth of Bacillus thuringiensis K2 and BTK2-T1, -T13, -T33 and -T40 got into stationary phase at 6 hour culture and then maintained it up to 48 hours without severe fluctuation. The production of beta-exotoxin from the strains, BTK2, BTK2-T1, -T13, -T17 and -T33 appeared at 6 hour culture and the amounts of the toxin were about 40 $\mu\textrm{g}$/$m\ell$ at 6 hour culture, approximately 70 $\mu\textrm{g}$/$m\ell$ at 12 hours, approximately 85$\mu\textrm{g}$/$m\ell$ from 24 hours to 48 hours. At 48 hour-culture, BTK2 produced 80 $\mu\textrm{g}$/$m\ell$ of beta-exotoxin (5.5$\times$10$^{8}$ cells/$m\ell$, BTK2-T13 produced 84 $\mu\textrm{g}$/$m\ell$ (4.3$\times$10$^{8}$ cells/$m\ell$), BTK2-T17 produced 87$\mu\textrm{g}$/$m\ell$ (1.4$\times$10$^{8}$ cells/$m\ell$), and BTK2-T33 produced 84 $\mu\textrm{g}$/$m\ell$ (4.9$\times$10$^{8}$ cells/$m\ell$). All other serotypes also produced beta-exotoxin. At 48 hour culture, BTK-37 produced 88$\mu\textrm{g}$/$m\ell$ (6.1$\times$10$^{8}$ cells/$m\ell$), BTK-35 produced 81 $\mu\textrm{g}$/$m\ell$), and the rest of them produced less than 70 $\mu\textrm{g}$/$m\ell$. To check the toxicity of beta-exotoxin and B. thuringiensis, the cultured media with microorganisms were inoculated to mice by per os, intraperiloneal, subcutaneous and intracerebral injection, and nasal cavity inoculation for 30 days. However, the toxin did not kill all of the treated mice.

• Clinical and Experimental Pediatrics
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• v.45 no.2
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• pp.183-191
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• 2002

Purpose : X-linked agammaglobulinemia(XLA) is an immunodeficiency caused by abnormalities in Bruton's tyrosine kinase(Btk), and is characterized by a deficiency of peripheral blood B cells. We studied the cytoplasmic expression of Btk protein and analyzed the Btk gene in peripheral blood mononuclear cells from two siblings and one cousin with XLA, as well as additional family members. Methods : Btk protein expression was analyzed by flow cytometry. Isolation of the coding sequence of the Btk gene was performed by amplification using the reverse transcription-polymerase chain reaction(RT-PCR) technique. Sequence alterations were screened by the single-stranded conformation polymorphism(SSCP) method and characterized by standard sequencing protocols. Results : Cytoplasmic expression of Btk protein in monocytes was not detected in three patients with XLA. In addition, Btk protein analysis clearly showed cellular mosaicism in monocytes from four obligate carriers, findings further supported by SSCP. A single base pair mutation(T to C) in Btk-exon three, which encodes the PH domain, was identified in four XLA patients. A diagnostic sequencing analysis was established to detect heterozygotic pattern in 4 carrier females. Furthermore, we found significant clinical heterogeneity in individuals with the same gene mutation. Conclusion : The implicating genetic alteration provided valuable clues to the pathogenesis of XLA in Korea and the flow cytometric analysis was suggested as a useful tool for rapid detection of XLA patients and carriers. The present study has identified a genetic mutation in the Btk coding region and demonstrated heterogeneity in clinical manifestations among patients with the same mutation. A flow cytometric analysis was found to be informative in establishing a deficiency of Btk protein in both patients and carriers and is recommended as a frontline procedure in the molecular diagnosis and work-up of XLA.

• Microbiology and Biotechnology Letters
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• v.13 no.3
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• pp.315-319
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• 1985

Delta-endotoxin crystals of B. thuringiensis var. kurstari and B. thuringiensis var. israelensis were purified by NaBr density gradient centrifigation and the wet weight of the BTK endotoxin was approximately 23.79％ of the cell wet weight and that of BTI was 25％. The shape of BTK crystal was bipyramidal, whose size was 1.7${\mu}{\textrm}{m}$ $\times$ 0.9${\mu}{\textrm}{m}$ and that of BTI was a spheroid, whose size was about 1.6$\times$0.45${\mu}{\textrm}{m}$. The molecular weight of BTK crystal protein was approximately 134,000 daltons and that of BTI was about 128,000 daltons.

• Clinical and Experimental Pediatrics
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• v.45 no.3
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• pp.302-310
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• 2002

Purpose : X-linked agammaglobulinemia(XLA) is an immunodeficiency caused by abnormalities in Bruton's tyrosine kinase(Btk), and is characterized by a deficiency of peripheral blood B cells. We studied cytoplasmic expression of Btk protein and analyzed the Btk gene in peripheral blood mononuclear cells(PBMC) from three XLA families in Korea. Methods : Heparinized venous blood samples were collected from four XLA patients and additional family members in three unrelated XLA families. Mononuclear cells were separated from their blood and the intracellular Btk protein was characterized by a flow cytometry. The mutation analysis was performed using direct sequencing. Results : Cytoplasmic expression of Btk protein in monocytes was not detected in the patients with XLA. We observed a novel deletion and two point mutations within introns(intron 1 and intron 18) resulting in alternative splicings. In XLA family 2, a 980 bp deletion(from intron 9+191 T to intron 10-215 C) including exon 10 was found in patient P2. He was the only sporadic case in this study, because his mother and brother showed a normal Btk expression by flow cytometry. Conclusion : These identified genetic alterations support the molecular heterogeneity of Btk gene in XLA disease. Additionally, by means of flow cytometric analysis, we diagnosed three hypogammaglobulinemia patients as XLA. Advancements in diagnostic methods has facilitated a prompt and definite diagnosis of this disease.

• Microbiology and Biotechnology Letters
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• v.26 no.4
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• pp.369-372
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• 1998

Microbial insecticide formulations were prepared by liquid and semi-solid fermentations using Bacillus thuringiensis subsp. kurstaki, HL-106 (BTK-HL106), B. thuringiensis subsp. israelensis HL-63 (BTI-HL63) and B. sphaericus 1593 (BS-1593) strains. The liquid fermentation medium contained molasses 2%, dextrose 1.5%, peptone 2%, D-xylose 0.025%, CaCl$_2$ 0.1%, K$_2$HPO$_4$ 0.1%, KH$_2$PO$_4$ 0.1%, MgSO$_4$$.$7H$_2$O 0.03%, FeSO$_4$$.$7H$_2$O 0.002%, ZnSO$_4$$.$7H$_2$O 0.02%. The composition of the semi-solid fermentation medium was rice bran 45.2%, zeolite 31%, yeast powder 0.02%, corn powder 5%, dextrose 3%, lime 0.3%, NaCl 0.06%, CaCl$_2$ 0.02%, and H$_2$O 15.42%. Insecticide formulations produced in the liquid fermentation named BTK-HL106, BTI-HL63 and BS-1593 pesticides and those in the semi-solid fermentation were designated as BTK-HL106-1, BTI-HL63-1 and BS-1593-1 pesticides, respectively. The number of spore (endotoxin crystals) was 2.65${\times}$10$\^$9/ spores per $m\ell$ in the BTK-HL106 and 3.5${\times}$10$\^$10/ in the BTK-HL106-1 3.8${\times}$10$\^$9/ spores in the BTI-HL63 and 7.0${\times}$10$\^$10/ in the BTI-HL63-1, and 7.5${\times}$10$\^$9/ in the BS-1593 and 1.4${\times}$10$\^$10/ in the BS-1593-1. The spores in the BS-1593 formulation was produced two times more than the other formulations. The spores in the BTI-HL63-1 were contained twice than those in the BTK-HL106-1, and five times than those in the BS-1593-1. The results indicated that spore (endotoxin crystals) productions in the semi-solid fermentation increased about ten times than those in the liquid fermentations. $LC_{50}$s of the BTI-HL63 and BS-1593 were 4.5 $\mu\textrm{g}$, and those of the BTI-HL63-1 and BS-1593-1 were 1.5 $\mu\textrm{g}$. $LC_{50}$ of the BTK-HL106 was 1.5 mg and that of the BTK-HL106-1 was 0.9 mg. The $LC_{50}$s of the formulations in the semi-solid fermentations showed about two to three times higher than those in the liquid fermentations.

• Pediatric Infection and Vaccine
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• v.24 no.3
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• pp.152-159
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• 2017

Purpose: X-linked agammaglobulinemia (XLA) is a primary immunodeficiency caused by mutations in the Bruton's tyrosine kinase (Btk) gene. The aim of this study was to investigate the clinical manifestations, molecular features, and treatment status of XLA in Korean patients at Seoul National University Children's Hospital. Methods: Fourteen Korean boys with XLA showing serum agammaglobulinemia, non-detectable to less than 2% of peripheral B-cells, and mutation of the Btk gene were enrolled. We observed the clinical features, laboratory findings, status of treatment, and complications in these XLA patients. Results: All XLA patients had a history of recurrent bacterial infections before diagnosis, and 20% of them had a neutropenia. Of the XLA patients 35.7% had a family history of XLA and 75% of their mothers were carriers. Btk gene analysis showed variable gene mutations in Xq22 including 9 amino acid substitutions, 3 frameshifts, 1 premature stop codon, and 1 splice defect. After intravenous immunoglobulin replacement therapy, infection episodes decreased, but complications such as bronchiectasis and chronic sinusitis remained. Conclusions: In patients less than 4 years of age with recurrent infection, analysis of serum gamma globulin levels and the Btk gene are recommended for the early diagnosis of XLA and for the appropriate prevention of recurrent infection.

• Journal of Microbiology and Biotechnology
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• v.4 no.4
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• pp.322-326
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• 1994

Bacillus thuringiensis strain BT-14 was isolated from alfalfa dust in Korea. The strain BT-14 produced one bipyramidal crystal and one spore in the cell. The biochemical characteristics of the strain BT-14 were similar to those of Bacillus thuringiensis subsp. kurstaki HD-l. Examination of its antibiotic resistance revealed that while the strain BT-14 was less resistant than BTK HD-l to ampicillin, gentamycin, neomycin and tobramycin, it was more resistant to amikacin than BTK HD-l. The $\delta$-endotoxin crystal of strain BT-14 consisted of a single protein with a high molecular weight of ca 135 KD on a 10% SDS-PACE. The strain BT-14 contained at least nine different plasmids with sizes of 2.9, 5.3, 5.8, 6.2, 9.4, 15.1, 18.1, 23.1 and 79 Kb. In insect bioassay, the isolated strain BT-14 showed lethality of 67% against Plutella xylostella larvae at dilution of 5$\times$$l0^{-4}$ (5$\times$l0 to 3$\times$$l0^2$ spores/ml), which is, almost equivalent to that of BTK HD-l.

• Tuberculosis and Respiratory Diseases
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• v.65 no.3
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• pp.207-211
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• 2008

X-linked agammaglobulinemia is the most common type of primary immunodeficiency disorder. Mutation ofthe cytoplasmic tyrosine kinase gene, Btk (Bruton's tyrosine kinase), is known to be the etiology of X-linked agammaglobulinemia. The patients with this disease manifest a B-cell deficiency and low levels of serum immunoglobulin; due to the deficient antibodies, they suffers from recurrent upper and lower respiratory infections. We report here a 24-year-old male with an initial clinical impression of recurrent pneumonia and bronchiectasis. The patient presented with marked pan-hypogammaglobulinemia and the absence of circulating B-lymphocytes on the immunologic study, and he carried a splicing mutation of intron 2 in the Btk gene (IVS2 -3C>G).

• Clinical and Experimental Pediatrics
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• v.59 no.sup1
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• pp.49-52
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• 2016

X-linked agammaglobulinemia (XLA) is a hereditary humoral immunodeficiency that results from Bruton's tyrosine kinase (BTK ) gene mutations. These mutations cause defects in B-cell development, resulting in the virtual absence of these lymphocytes from the peripheral circulation. Consequently, this absence leads to a profound deficiency of lg all isotypes, and an increased susceptibility to encapsulated bacterial infections. A 15-month-old Korean boy presented with recurrent sinusitis and otitis media after 6 months of age, and had a family history of 2 maternal uncles with XLA. Laboratory tests revealed a profound deficiency of Ig isotypes, and a decreased count of $CD19^+$ B cells in the peripheral circulation. Based on his family history and our laboratory test results, he was diagnosed with XLA. We performed BTK gene analysis of peripheral blood samples obtained from family members to confirm the diagnosis. Mutational analysis revealed a novel hemizygous frameshift mutation (c.82delC, p.Arg28Alafs*5), in the BTK gene. His mother and maternal grandmother were heterozygous carriers of this mutation and his two maternal uncles were hemizygous at the same position. After XLA diagnosis, intravenous immunoglobulin (400 mg/kg, monthly) treatment was initiated; recurrent sinusitis and otitis media were subsequently brought under control. To our knowledge, this is the first reported case of a Korean pedigree with a novel mutation in the BTK gene.