Inhibitory Effects of Selected Medicinal Plants on Bacterial Growth of Methicillin-Resistant Staphylococcus aureus.

Methicillin-resistant Staphylococcus aureus (MRSA) is a serious threat to global public health due to its capacity of tolerate conventional antibiotics. Medicinal plants are traditionally used to treat infectious diseases caused by bacterial pathogens. In the present study, 16 medicinal plants were screened for antibacterial activities to preselect more effective species. Ethanol extracts of selected medicinal plants (Caesalpinia sappan L., Glycyrrhiza uralensis Fisch., Sanguisorba officinalis L., and Uncaria gambir Roxb) were partitioned successively with different solvents (n-hexane, chloroform, ethyl acetate, 1-butanol, and water). Disc diffusion assay and broth microdilution were performed to evaluate the antibacterial activities of plant extracts and fractions against Staphylococcus aureus strains. Furthermore, the cytotoxicity of the extracts and fractions was determined against the human hepatoma (HepG2) and human lung carcinoma (A549) cell lines using a trypan blue exclusion method. A few extracts and fractions showed significant inhibitory effects on the bacterial growth of all tested strains, including multidrug-resistance (MDR) clinical isolates. The ethyl acetate fraction of C. sappan had the most potent effects with minimum inhibitory/bactericidal concentrations (MIC/MBC) of 31.2/62.5 µg/mL and showed low cytotoxicity with over 90% cell viability in both cells. Our results suggest that medicinal plants have considerable potential as alternatives to conventional antibiotics.

Korean Red Ginseng and Ginsenoside Rg3 Suppress Asian Sand Dust-Induced Epithelial-Mesenchymal Transition in Nasal Epithelial Cells.

Chronic rhinosinusitis (CRS) is characterized by chronic inflammation of the sinonasal mucosa with epithelial dedifferentiation toward the mesenchymal phenotype, known as the epithelial-mesenchymal transition (EMT). Asian sand dust (ASD) can induce nasal mucosal inflammation and cause the development of EMT. Korean red ginseng (KRG) and ginsenoside Rg3 have been used as traditional herbal medicines to treat various diseases. The aim of this study was to investigate their effect on ASD-induced EMT in nasal epithelial cells. Primary nasal epithelial cells were incubated with ASD with or without KRG or Rg3, and the production of transforming growth factor-ß1 (TGF-ß1) and interleukin (IL)-8 was measured. EMT markers were determined by RT-PCR, Western blot analysis, and confocal microscopy, and transcription factor expression by Western blot analysis. The effect on cell migration was evaluated using the wound scratch assay. Results showed ASD-induced TGF-ß1 production, downregulation of E-cadherin, and upregulation of fibronectin in nasal epithelial cells. KRG and Rg3 suppressed TGF-ß1 production (31.7% to 43.1%), upregulated the expression of E-cadherin (26.4% to 88.3% in mRNA), and downregulated that of fibronectin (14.2% to 46.2% in mRNA and 52.3% to 70.2% in protein). In addition, they suppressed the ASD-induced phosphorylation of ERK, p38, and mTOR, as well as inhibiting the ASD-induced migration of nasal epithelial cells (25.2% to 41.5%). The results of this study demonstrate that KRG and Rg3 inhibit ASD-induced EMT by suppressing the activation of ERK, p38, and mTOR signaling pathways in nasal epithelial cells.

Anti-Oxidant Activity and Dust-Proof Effect of Chitosan with Different Molecular Weights.

High molecular weight chitosan (HMWC) was degraded to prepare chitosan with different molecular weight based on the fenton reaction, which can produce aggressive OH-radicals produced from hydrogen peroxide in the presence of catalytic metal ions. The relative molecular weight, anti-oxidant activity, and fine dust removal effect of chitosan hydrolysates were elucidated to define their molecular weight and their potent biological activity. Our results demonstrate that chitosan hydrolysates derived from the hydrolysis of HMWC may possess significant free-radical scavenging activity as good anti-oxidants against the radical scavenging activity of DPPH and ABTS, respectively. Furthermore, chitosan hydrolysates can effectively eliminate fine dust, which may contain some particulate matter (PM) and unknown species of microorganisms from the air, suggesting that our data provide important information for producing air filters, dust-proof masks and skin cleaner for the purpose of human healthcare and well-being.

Potent biomedical applications of isolated polysaccharides from marine microalgae Tetraselmis species.

Microalgae Tetraselmis species were used to evaluate the biological characteristics of water-soluble polysaccharides (WSPs) as one of the significant bioactive substances (BAS) from these photosynthetic microalgae species. Compositional analysis of these BAS shows that they are mainly composed of WSPs along with negligible amount of proteins and lipids. WSPs were partially purified and characterized for their compositional, structural and biological properties such as antioxidant, tyrosinase inhibitory activity and antifungal activies. These WSPs showed the significant antioxidant, antifungal and tyrosinase inhibitory activities, respectively. The outcomes of this study demonstrated that WSPs can be the potent source of biological moieties for further investigations along with specific potent biological activities.

Natural Phytochemical and Visible Light at Different Wavelengths Show Synergistic Antibacterial Activity against Staphylococcus aureus.

As the risk of antibiotic-resistant bacteria increases, interest in non-antibiotic treatment is also increasing. Among the methods used in non-antibiotic therapy, natural antibiotics such as essential oils have disadvantages such as low efficiency. In the case of phototherapy, the light used for antibacterial activities has low penetration into the human body because of its short wavelength, making it of low medical utility. To solve this problem, this study aimed to determine conditions for enhancing the antibacterial activity of natural phytochemicals and visible light. Four natural phytochemical extracts that showed high antibacterial properties in previous studies were analyzed. Synergistic effects on antibacterial activity and cytotoxicity were determined when natural phytochemical extracts and visible light were simultaneously used. As a result, it was confirmed that the antibacterial activity increased by four times when Sanguisorba officinalis L. was irradiated with 465 nm for 10 min and 520 nm for 40 min, and Uncaria gambir Roxb. was irradiated with 465 nm for 10 min and 520 nm for 60 min compared to when Sanguisorba officinalis L. and Uncaria gambir Roxb. were used alone. The synergistic effect on antibacterial activity was independent of the absorption peak of the natural phytochemical extracts. In addition, in the case of natural phytochemical extracts with improved antibacterial activity, it was confirmed that the improvement of antibacterial activity was increased in inverse proportion to the light irradiation wavelength and in proportion to the light irradiation time. The antibacterial activity was enhanced regardless of antibiotic resistance. In the case of cytotoxicity, it was confirmed that there was no toxicity to A549 cells when treated with 465 nm, the shortest wavelength among the natural phytochemical extracts. These results show how to replace blue light, which has been underutilized due to its low transmittance and cytotoxicity. They also demonstrate the high medical potential of using natural phytochemical and visible light as a combination therapy.

The effect of Asian sand dust-activated respiratory epithelial cells on activation and migration of eosinophils.

UNLABELLED: CONTEST: Asian sand dust (ASD) contains various chemical and microbiological materials. ASD aggravate the inflammatory response of respiratory epithelial cells and symptoms of asthma. OBJECTIVE: To evaluate the inflammatory effects of ASD on the activation of bronchial epithelial cells and the effect on the activation and migration of eosinophils. MATERIALS AND METHODS: BEAS-2B cells were exposed to three forms of ASD: particles less than 10 µm in diameter (PM), dried sand dust (SD) and sand dust collected from the Gobi Desert (GB). Activation of the epithelial cells was determined using interleukin-6 (IL-6), IL-8, granulocyte macrophage colony-stimulating factor (GM-CSF), regulated on activation normal T expressed and secreted (RANTES), and eotaxin. Eosinophil migration was induced with bronchial epithelial cell conditioned medium. Eosinophils were stimulated with the ASDs and production of superoxide and eosinophil cationic protein was measured. RESULTS: PM and SD enhanced the production of IL-6, IL-8 and RANTES. However, only IL-6 production was significantly increased with GB. Conditioned medium stimulated PM and SD enhanced the migration of eosinophils. PM and SD strongly activated eosinophils. DISCUSSION AND CONCLUSIONS: ASD, which contains smaller particles and air pollutants, might exacerbate the inflammatory process of bronchial tissue and asthmatic symptoms with the production of inflammatory mediators and tissue eosinophilia.

Comparing the Phylogenetic Distribution of Multilocus Sequence Typing, Staphylococcal Protein A, and Staphylococcal Cassette Chromosome Mec Types in Methicillin-Resistant Staphylococcus Aureus (MRSA) in Korea from 1994 to 2020.

Staphylococcus aureus (S. aureus) bacteremia is one of the most frequent and severe bacterial infections worldwide. Methicillin-resistant Staphylococcus aureus (MRSA) is a serious human pathogen that can cause a wide variety of infections. Comparative genetic analyses have shown that despite the existence of a vast number of genotypes, genotypes are restricted to certain geographical locations. By comparing multilocus sequence typing (MLST) and SCCmec types from 1994 to 2020, the present study intended to discover which genotype genes were related to MRSA infections. MLST, Staphylococcus aureus protein A (spa), and SCCmec typings were performed to determine their relationship during those years. Results revealed that MRSA isolates in the Republic of Korea were distributed among all major staphylococcal cassette chromosome mec (SCCmec) types. The majority of SCCmec isolates belonged to SCCmec type II and type IV. The majority of MLST had the sequence type (ST) 72, 239, 8, or 188. By contrast, minorities belonged to ST22 (SCCmec IV), ST772 (SCCmec V), and ST672 (SCCmec V) genotypes. The SCCmec type was determined for various types. The spa type was dispersed, seemingly regardless of its multidrug resistance property. The MLST type was found to be similar to the existing typical type. These results showed some correlations between resistance characteristics and types according to the characteristics of the MLST types distributed, compared to previous papers. Reports on genotype distribution of MLST and SCCmec types in MRSA are rare. These results show a clear distribution of MLST and SCCmec types of MRSA from 1994 to 2020 in the Republic of Korea.

In Vitro Synergistic Inhibitory Effects of Plant Extract Combinations on Bacterial Growth of Methicillin-Resistant Staphylococcus aureus.

Methicillin-resistant Staphylococcus aureus (MRSA) is one of the most common pathogens of healthcare-associated infections. Medicinal plants have long been used in the traditional treatment of diseases or syndromes worldwide. Combined use of plant extracts could improve the effectiveness of pharmacological action by obtaining synergism, acting on multiple targets simultaneously, reducing the doses of individual components, and minimizing side effects. We aimed to investigate the synergistic inhibitory effects of selected medicinal plants (Caesalpinia sappan L. (CS), Glycyrrhiza uralensis Fisch. (GU), Sanguisorba officinalis L. (SO), and Uncaria gambir Roxb. (UG)) on the bacterial growth of MRSA and its clinical isolates. SO and UG extracts generated the best synergistic interaction as adjudged by checkerboard synergy assays. MICs of the individual extracts decreased 4-fold from 250 to 62.5 µg/mL, respectively. The SO + UG combination was further evaluated for its effects on bacterial growth inhibition, minimum bactericidal/inhibitory concentration (MBC/MIC) ratio, and time-kill kinetics. The results indicate that the SO + UG combination synergistically inhibited the bacterial growth of MRSA strains with bactericidal effects. SO + UG combination also exhibited more potent effects against clinical isolates. In multistep resistance selection experiments, both standard and isolates of MRSA showed no resistance to the SO + UG combination even after repeated exposure over fourteen passages. Our data suggest that using plant extract combinations could be a potential strategy to treat MRSA infections.

Asian sand dust enhances rhinovirus-induced cytokine secretion and viral replication in human nasal epithelial cells.

CONTEXT: Asian sand dust (ASD) originating in the arid deserts of Mongolia and China causes annual severe air pollution events in the Asia-Pacific area, including Korea, Japan, and China. ASD is thought to impact public health by aggravating or inducing respiratory illness. Among the most common respiratory illnesses is the common cold caused by rhinovirus (RV) infection. To date, however, the impact of ASD on RV infection has not been studied. OBJECTIVE: In this study, we investigated the effect of ASD on RV infection in human nasal epithelial cells. METHODS: Primary human nasal epithelial cells grown at an air-liquid interface were treated with ASD and/or RV. After RV infections were confirmed using semi-nested reverse transcription-polymerase chain reaction (RT-PCR), mRNA expression and protein secretion of the inflammatory cytokines interferon-γ (IFN-γ), interleukin-1ß (IL-1ß),IL-6, and IL-8, indicators of the severity of RV-induced inflammation, were measured by real-time PCR and enzyme-linked immunosorbent assays. Viral titer was also assayed by culturing viruses to compare viral replication between RV-only and ASD-plus-RV groups. RESULTS: ASD significantly increased RV-induced IFN-γ, IL-1ß, IL-6, and IL-8 mRNA levels and protein secretion in primary nasal epithelial cells. In addition, ASD caused a significant increase in RV replication. CONCLUSIONS: Our results suggest that ASD may potentiate common cold symptoms associated with RV infection not only by enhancing IFN-γ, IL-1ß, IL-6, and IL-8 secretion, but also by increasing viral replication.

Asian dust and titanium dioxide particles-induced inflammation and oxidative DNA damage in C57BL/6 mice.

Inhaled particulate matter (PM) might influence many adverse health effects in human body, including increased exacerbations of pulmonary and cardiovascular diseases. In this study, we examined the associations between PM and pulmonary adverse effects. Two types of particles, Asian dust (AD) and titanium dioxide (TiO(2)), were administered intratracheally to C57BL/6 mice. The mice were exposed to saline and saline suspensions of 20 mg/kg of AD, TiO(2) particles twice a week for 12 weeks. Following exposure with these particles, the lungs were analyzed histopathologically by hematoxylin and eosin (H&E) and Masson's trichrome (MT) staining. Oxidative injuries were determined by immunohistochemistry (IHC) for 8-oxoguanine in the lungs and Comet assays in peripheral blood mononuclear cells (PBMCs) of C57BL/6 mice. Mice exposed to AD and TiO(2) showed significant inflammatory changes and oxidative damages in the lungs as compared with the control group. DNA damage in PBMCs was also increased significantly in AD and TiO(2)-exposed mice. However, lung fibrosis was minimal and there was no significant difference between PM exposed and control mice. Exposure to AD and TiO(2) particles-induced similar inflammatory damages in the lungs and elicited oxidative DNA damage in the PBMCs.

Novel spa and Multi-Locus Sequence Types (MLST) of Staphylococcus Aureus Samples Isolated from Clinical Specimens in Korean.

Staphylococcus aureus bacteremia is one of the most frequent and severe bacterial infections worldwide. The increased incidence of S. aureus infections with a diverse pattern of S. aureus protein A (spa) types across different geographic regions is a global challenge. This study investigated a novel spa type of methicillin-resistant S. aureus in a clinically isolated specimen. A total of 109 clinical S. aureus samples were subjected to 19 sets of antimicrobial susceptibility testing using the Kirby-Bauer disk diffusion method. Molecular typing was performed with S. aureus protein A (spa) and multi-locus sequence types (MLST) via polymerase chain reaction and sequencing. The methicillin-resistant S. aureus samples in our study accounted for 55.05% (60/109) of the total. A novel spa type was detected in five (5/60) strains. This gh22 isolate was identified in antimicrobial susceptibility tests of 15 kinds of antibiotics. Antibiotic resistance genes included mecA, TEM, aac(6')-aph(2"), ermA, and tetM. Eleven S. aureus samples were classified as t2460, t338, t324, t693, five unknown spa types (new spa types), and undefined MLST (novel MLST). We report a high prevalence rate of t2460 methicillin-resistant S. aureus samples in our country. Additionally, novel spa gh22, MLST ST4613, and clonal compact CC5-type strains (T1:M1:B1:B1:M1:E1:K1, r26:r17:r34:r34:r17:r13:r16, mlst;1:4:1:4:559:495:10) showing multidrug resistance were identified among S. aureus samples.

Effect of exposure to Asian sand dust-Particulate matter on liver Tenascin-C expression in human cancer cell and mouse hepatic tissue.

Asian Sand Dust-Particulate Matter (ASD-PM) aerosol brings large amounts of wind-eroded soil particles containing high concentrations of metallic components caused by industrialization and vehicles. Proinflammatory and cytotoxic cytokines trigger local inflammatory responses and cause a systematically high incidence of cardiovascular and other diseases. Tenascin C (Tn-C) is known to be expressed in damaged tissue or in a developmental stage of tissue. In this study, we examined the expression of Tn-C and Fibronectin in human cancer-cell lines and in liver tissue of mice treated with ASD-PM to investigate the inflammatory and cell-damage effects of ASD-PM. In our in vivo study, mice were intratracheally instilled with saline suspensions of ASD-PM particles. Instillation of these particles was repeated twice a week for 12 weeks and the liver tissues were stained with hematoxylin, eosin, and Masson's trichrome, and we carried out an IF. Tn-C expression in liver tissues was detected by RT-PCR and western blot analysis. In the results, the expression of Tn-C increased in a dose-dependent manner in both RNA and Immunofluorescence assay (IF). In our in vitro study, A549 and Hep3B cell lines were incubated in culture media with Transforming Growth Factor-Beta1(TGF-ß1) and ASD-PM. Immunofluorescence microscopy images showed a two times stronger expression of fluorescence in the ASD-treated group than in that treated with TGF-ß1. They also showed a stronger expression of Tn-C in proportion to the concentration of ASD-PM. We confirmed that ASD-PM when inhaled formally migrated to other organs and induced Tn-C expression. ASD-PM containing metals causes expression of Tn-C in liver tissue in proportion to the concentration of ASD-PM.

Rarity of TLR4 Asp299Gly and Thr399Ile polymorphisms in the Korean population.

PURPOSE: Activation of the innate immune system and chronic low-grade inflammation are thought to be involved in the pathogenesis of atherosclerosis and also thought to be associated with type 2 diabetes and its complications. As a receptor for bacterial lipopolysaccharide and heat-shock proteins, Toll-like receptor 4 (TLR4) is one of the central regulators of the immune response. Recent studies have reported an association between TLR4 polymorphisms and diabetes and its complications in Caucasian populations. MATERIALS AND METHODS: In this study, we analyzed the association between TLR4 gene polymorphisms in patients with features of type 2 diabetes and healthy controls in Korea. Two polymorphisms of the TLR4 gene (Asp299Gly and Thr399Ile) were examined in 225 diabetic patients and 153 healthy controls using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and single-strand conformation polymorphism (SSCP). RESULTS: No Asp299Gly or Thr399Ile mutations were detected in any of the 378 subjects. Seven subjects from each group who had slightly different SSCP patterns were selected for sequencing, but we found no TLR4 polymorphisms on Exon3. The Asp299Gly and Thr399Ile TLR4 gene polymorphisms were absent in both groups, which was similar to the results for Japanese and Chinese Han subjects. CONCLUSION: Our data and other Asian data suggest that a racial difference can be found in the frequency of the TLR4 polymorphism.

Tumor-associated carbonic anhydrases are linked to metastases in primary cervical cancer.

PURPOSE: Carbonic anhydrase IX (CA9) has emerged as an important surrogate marker for hypoxia in solid tumors. CA12 shares a role with CA9 in acidification of micromilieu but it is less strictly regulated by hypoxia than CA9. In this study, we investigated expression of CA9 and CA12 mRNA in primary cervical cancer. We also examined whether CA9 expression can be an indicator of reoxygenation of tumor by measuring its mRNA expression during fractionated radiotherapy. METHODS: Tumor tissues were obtained from 59 patients with uterine cervical cancer who underwent radiotherapy, and a second biopsy was taken after patients had received either 10 or 20 Gy of radiation. The follow-up period ranged from 2.4 to 75 months (median=23 months). The ratio of CA9 and beta-actin mRNA expression was determined both pre- and during radiation treatment by RT-PCR. RESULTS: CA9 and CA12 mRNA expression was detected in 62.7 and 88.1% of tumors (i.e. patients), respectively, and co-expression was observed in 61% of patients. Multivariate analysis revealed that CA9 expression was the most significant factor associated with metastasis-free survival (P=0.008, hazard ratio 34.8), whereas CA12 mRNA expression was linked to a lower risk of metastasis (P=0.007, hazard ratio of 0.07). Tumor CA9 expression was not altered following either 10 or 20 Gy of radiotherapy. CONCLUSION: The strong correlation between CA9 expression and metastasis suggests that CA9 expression might be an important indicator for identifying patients who require more aggressive systemic therapy.

Dexamethasone inhibits hypoxia-induced epithelial-mesenchymal transition in colon cancer.

AIM: To elucidate the effects of dexamethasone on hypoxia-induced epithelial-to-mesenchymal transition (EMT) in colon cancer. METHODS: Human colon cancer HCT116 and HT29 cells were exposed to normoxic (21%) and hypoxic (1%) conditions. First, the effect of dexamethasone on cell viability was examined by MTT cell proliferation assay. In order to measure the expression levels of EMT markers (Snail, Slug, Twist, E-cadherin, and integrin αVß6) and hypoxia-related genes [Hypoxia-inducible factor-1α (HIF-1α) and vascular endothelial growth factor (VEGF)] by dexamethasone, quantitative real-time polymerase chain reaction and western blot analysis were performed. Furthermore, the morphological changes of colon cancer cells and the expression pattern of E-cadherin by dexamethasone were detected through immunocytochemistry. Finally, the effects of dexamethasone on the invasiveness and migration of colon cancer cells were elucidated using matrigel invasion, migration, and wound healing migration assays. RESULTS: Under hypoxia, dexamethasone treatment inhibited HIF-1α protein level and its downstream gene, VEGF mRNA level in the colon cancer cell lines, HCT116 and HT29. In addition, the presence of dexamethasone down-regulated the mRNA levels of hypoxia-induced Snail, Slug, and Twist, all transcriptional factors of EMT, as well as hypoxia-induced integrin αVß6 protein level, a well-known EMT marker for colon cancer cells. Furthermore, reduced E-cadherin in hypoxic condition was found to be recoverable by treating with dexamethasone in both colon cancer cell lines. Similarly, under hypoxic conditions, dexamethasone restored the growth pattern and morphological phenotype reminiscent of colon cancer cells grown under normoxic conditions; dexamethasone blocked the migration and invasion of both colorectal cancer cell lines in hypoxia. CONCLUSION: Our study suggested that dexamethasone has inhibitory effects on cell migration and invasion by suppressing EMT of colon cancer cell lines in hypoxic condition.

Expression of cathelicidin in human salivary glands.

BACKGROUND: Salivary secretions play a critical role in maintaining oral health via innate host defense mechanisms and secretion of secretory IgA. One of the antimicrobial peptides, LL-37, is the only cathelicidin protein that has yet been identified in humans. Cathelicidins are a family of peptides thought to provide an innate defensive barrier against a variety of potential microbial pathogens. OBJECTIVES: To examine the expression of cathelicidin in human salivary glands and to investigate up-regulation of cathelicidin in inflammatory conditions. DESIGN: Reverse transcriptase-polymerase chain reaction and immunohistochemical staining were performed on 20 salivary gland tissues, 10 from normal salivary glands and 10 from glands with chronic sialadenitis. RESULTS: Cathelicidin messenger RNA transcripts were detected in the tissues from the normal salivary glands and the glands with chronic sialadenitis. The level of cathelicidin messenger RNA in glands with chronic sialadenitis was significantly increased compared with that in normal salivary glands. Cathelicidin protein was expressed in the glandular epithelium of the normal salivary glands and the glands with chronic sialadenitis. CONCLUSION: The results indicate that cathelicidin might play an important role in innate host defense of human salivary glands.

Antimicrobial peptide LL-37 is upregulated in chronic nasal inflammatory disease.

OBJECTIVE: LL-37 is one of the antimicrobial peptides and the only member of the cathelicidin family identified so far in humans. We attempted to find a correlation between LL-37 peptide and inflammation of the nasal mucosa. MATERIAL AND METHODS: Nasal mucosa specimens were obtained from 15 chronic infective rhinitis patients and 6 normal controls. Immunohistochemical staining was used to examine the localization of LL-37 and reverse transcriptase polymerase chain reaction (RT-PCR) was used to determine the levels of LL-37, IL-1beta and IL-8 in tissue. RESULTS: LL-37 peptide was primarily localized in the surface of the epithelia, in the serous and mucous cells of the submucosal glands and in stromal inflammatory cells. The number of LL-37 immunoreactive cells in inflammatory nasal mucosa was significantly increased compared with normal nasal tissue. Using RT-PCR, LL-37 mRNA was detected in 3/6 normal turbinate samples but in all cases with inflammatory nasal tissues. IL-1beta and IL-8 transcripts exhibited a similar pattern to that of LL-37. CONCLUSIONS: We suggest that LL-37 is one of the antimicrobial peptides found in human nasal mucosa and that it participates in the innate immune system of the nasal mucosa.

[Polymorphism in codons 10 and 25 of the transforming growth factor-beta1 gene in Korean population and in patients with liver cirrhosis and hepatocellular carcinoma].

BACKGROUND/AIMS: The genetic polymorphism of transforming growth factor-beta1 (TGF-beta1) at codons 10 and 25 which influences the production of TGF-beta1 is related to fibrogenesis in the lung and liver. We evaluated the genetic polymorphism at codons 10 and 25 in controls and in patients with liver cirrhosis (LC) and hepatocellular carcinoma (HCC). METHODS: Blood samples were collected from controls (n=35), patients with LC (n=64), and HCC (n=49). Genomic DNA was isolated and polymerase chain reaction (PCR) was done for a segment including codons 10 and 25. The results of direct sequencing for PCR products were compared between the controls and the patients. RESULTS: There was no genetic polymorphism at codon 25 and three types of genetic polymorphism at codon 10. The leucine homozygous genotype (CTG/CTG) at codon 10 was more common in patients with LC than the controls (p=0.01) and especially in patients with LC caused by HBV (p=0.004). The polymorphism at codons 10 in patients with HCC was similar to the controls. However, leucine homozygous genotype was more common in patients with HCC of uninodular morphology than those of massive morphology (p=0.007). CONCLUSIONS: The genetic polymorphism of TGF-beta1 at codon 10 might be associated with LC and morphology of HCC. The potential usefulness of TGF-beta1 genotyping needs further studies in large scale.

Bacillus thuringiensis: a specific gamma-cyclodextrin producer strain.

An anaerobic microbial isolate Bacillus species, designated B. thuringiensis GU-2, was isolated from soil as a specific γ-cyclodextrin (CD) producer strain in alkaline medium under anaerobic conditions. The optimum pH and temperature for bacterial growth and γ-CD production were estimated to be pH 8.5 and 37°C in the presence of 1.0% starch substrate, respectively. A high purity yield >95% of γ-CD from the total CD yield in the reaction mixture was obtained from starch that was supposed to be converted by gamma-cyclodextrin glycotransferase, tentatively named as γ-CGTase. The maximum γ-CGTase activity was estimated at 2.45U/mL under optimized condition. This is the first report demonstrating the generation of a specific γ-cyclodextrin (CD) producer strain by the action of a γ-CGTase under anaerobic conditions.