• Title/Summary/Keyword: cells$

Search Result 48,448, Processing Time 0.068 seconds

Cold Shock Response and Low Temperature Stable Transcript of DEAD-box RNA Helicase in Bacillus subtilis (DEAD-box RNA Helicase 유전자가 결핍된 Bacillus subtilis의 저온 충격 반응성과 저온 안정성 전사물)

  • Oh, Eun-Ha;Lee, Sang-Soo
    • Korean Journal of Microbiology
    • /
    • v.47 no.4
    • /
    • pp.289-294
    • /
    • 2011
  • We investigated the cold shock sensitivity of DEAD-box RNA helicase gene deleted strains of in Bacillus subtilis CU1065. To understand cold shock effects, cells were cultivated at $37^{\circ}C$ to log phase ($O.D_{600}$=0.5-0.6) and then temperature was shifted to $15^{\circ}C$. Cold shock slow down the growth rate of wild type and deleted strains of DEAD-box RNA helicase gene (ydbR, yfmL, yqfR, deaD). The growth rate of ydbR deleted strain is 5 times severely reduced compared to that of wild type strain (CU1065). But the growth rate of other three (yfmL, yqfR, deaD) deleted strains is nearly equal to the growth rate of wild type. Compared to $37^{\circ}C$, the amount of ydbR and yqfR mRNA transcripts are increased at the growth temperature of $15^{\circ}C$. On the other hands the mRNA transcripts of yfmL and deaD are not changed at both conditions of $37^{\circ}C$ and $15^{\circ}C$. Upon cold shock treatment ydbR mRNA transcript is clearly increased. After treatment of rifampicin (bacteria transcription inhibitor) the amount of ydbR mRNA was measured. Temperature shift from $37^{\circ}C$ to $15^{\circ}C$ and rifampicin treatment showed slowly decay of ydbR mRNA. But at $37^{\circ}C$ and rifampicin treatment ydbR mRNA is rapidly reduced. These results showed that cold shock induction of ydbR mRNA resulted from the stability of ydbR mRNA and not from the transcription induction of ydbR. In relation to these results, we found the cold box element of csp (cold shock protein gene) in 5' untranslated region of ydbR gene. Cold shock induction of ydbR is caused by the stability of ydbR mRNA like the stability of csp mRNA.

Cold shock sensitive growth of Bacillus subtilis mutants deleted for genes involved in fatty acid synthesis (지방산 생합성 관련 유전자 결손 Bacillus subtilis 균주들의 저온충격 민감성 생장)

  • Kim, Do Hyung;Lee, Sang Soo
    • Korean Journal of Microbiology
    • /
    • v.54 no.1
    • /
    • pp.9-17
    • /
    • 2018
  • To investigate the role of bkdR, sigL, yplP, and des genes which were known to be involved in fatty acid synthesis and sensitive at low temperature, deletion mutants of Bacillus subtilis CU1065 and JH642 were constructed. To determine the low temperature sensitivity of these genes, we compared the growth curves of cells at $37^{\circ}C$ and $15^{\circ}C$. At $37^{\circ}C$, wild type and deletion mutants showed almost similar growth but only bkdR deletion strain at $15^{\circ}C$ showed very slow growing compared with wild type. At $15^{\circ}C$ sigL and yplP deletions were somewhat slower or similar to those of wild type strain. Double and triple mutants for bkdR, sigL, yplP deletions were constructed and grown at $20^{\circ}C$ in LB agar to investigate cold sensitive growth. Double or triple deletions including bkdR deletion showed cold sensitive growing. In order to identify more clearly cold sensitive growth, the experiments were carried out under cold shock conditions in which the temperature was lowered from $37^{\circ}C$ to $15^{\circ}C$ at the point of 0.4 optical densities at 600 nm. In these cold shock experiments, only bkdR deletion showed significantly lower growing and additional des deletion increases cold sensitivity. The bkdR activates the bkd operon, which catabolized isoleucine, valine and leucine, amino acids and produce precursors for the synthesis of branched fatty acids. At cold shock growing of bkdR deletion strain, isoleucine recovered cold sensitivity of bkdR deletion but valine did not restore cold sensitivity. Isoleucine is used as a precursor for the synthesis of anteiso-branched fatty acids. On the other hand, valine is used as a precursor for the synthesis of iso-branched fatty acids. This indicates that anteiso-branched fatty acid plays an important role at the cold shock condition.

Histopathological and Serological Observations on Experimental Anisakiasis of Rabbits (실험적 토끼 아니사키스증에서의 조직학적 병변과 혈청 항체가의 경시적 변동 양상)

  • 홍성태;이손형
    • Parasites, Hosts and Diseases
    • /
    • v.25 no.2
    • /
    • pp.168-180
    • /
    • 1987
  • This study was performed to observe histopathological changes and serological reactions in chronic anisakiasis of rabbits. Each rabbit was infected per os with 30 larvae of Anisakis type I. Their sera were collected chronologically and the rabbits were killed for histopathological examination, 3, 13, 20, 30, 60, 90 and 150 days after the infection. The results were summarized as below. 1. Most of the larvae were recovered from the stomach, but a few from the omentum, intestine, mesentery and abdominal wall. The recovery rates and distribution of worms by organ were not differed by duration of infection. 2. Histologically the lesion was abscess type on 13 days, i.e., the dead worms were surrounded by fibrinous exudate, histiocytes and thick zone of numerous inflammatory cells. After 30 days, histiocytes were found to invade the worms and the lesion was changing into abscessgranulomatous type. Also a calcified worm was found on the 30th day. After then the worms were observed to be dissolved slowly until 90 days. On 150 day, only one calcified worm was observed. 3. The levels of serum IgG antibody by ELISA reached their maximum 30 days after the infection. After then, it decreased slowly until 150 days after the infection. Above serological and histopathological findings indicated that antigenic stimulation from degenerating Anisakis larvae was the greatest during the first 30 days after infection. This period was corresponding with the beginning of worm resolution or calcification. Serologic test by ELISA would be a valuable tool for confirming chronic anisakiasis.

  • PDF

Usability of Urinary Reagent Strips in Diagnosis of Meningitis (뇌수막염의 진단에서 뇨검사 시험지를 이용한 뇌척수액 검사의 유용성)

  • Jeong, Jin Yeong;Park, Sang Kyu
    • Pediatric Infection and Vaccine
    • /
    • v.4 no.2
    • /
    • pp.265-270
    • /
    • 1997
  • Purpose: Rapid diagnosis and prompt treatment remain the cornerstone of management of patients with meningitis. Examination of cerebrospinal fluid(CSF) for glucose, protein, cells, and organisms is necessary for accurate diagnosis, but in many parts of the world facilities do not exist to do so. We tested CSF samples from 69 children with suspected meningitis with urinary reagent strips and analyzed the results to know the usability of urinary reagent strips in diagnosis of meningitis. Methods: 69 CSF samples obtained from children with suspected meningitis were analysed. Each sample was divided into two; one was sent to the laboratory for routine diagnostic evaluation, and the other was tested with the reagent strip(Combur10 Test M). Laboratory values of CSF glucose, protein, and leucocytes for each CSF sample were plotted against the corresponding reagent-strip category. Results: There was good agreement between the dipstick and laboratory values for CSF protein and cell count but not for glucose. Conclusion: Rapid diagnosis of meningitis could be made with three component of the Combur10 test strip. It is easy to do and would be of particular value to those working in parts of the world where no laboratory facilities exist.

  • PDF

Effects of Oxidative DNA Damage and Genetic Polymorphism of the Glutathione Peroxidase 1 (GPX1) and 8-Oxoguanine Glycosylase 1 (hOGG1) on Lung Cancer (GPX1 및 hOGG1 유전자다형성에 따른 유전자의 산화적 손상 및 폐암 발생 위험도 평가)

  • Lee, Chul-Ho;Lee, Kye-Young;Choe, Kang-Hyeon;Hong, Yun-Chul;Noh, Sung-Il;Eom, Sang-Yong;Ko, Young-Jun;Zhang, Yan-Wei;Yim, Dong-Hyuk;Kang, Jong-Won;Kim, Heon;Kim, Yong-Dae
    • Journal of Preventive Medicine and Public Health
    • /
    • v.39 no.2
    • /
    • pp.130-134
    • /
    • 2006
  • Objectives : Oxidative DNA damage is a known risk factor of lung cancer. The glutathione peroxidase (GPX) antioxidant enzyme that reduces hydrogen peroxide and lipid peroxides plays a significant role in protecting cells from the oxidative stress induced by reactive oxygen species. The aim of this case-control study was to investigate effects of oxidative stress and genetic polymorphisms of the GPX1 genes and the interaction between them in the carcinogenesis of lung cancer. Methods : Two hundreds patients with lung cancer and 200 age- and sex-matched controls were enrolled in this study. Every subject was asked to complete a questionnaire concerning their smoking habits and their environmental exposure to PAHs. The genotypes of the GPX1 and 8-oxoguanine glycosylase 1 (hOGG1) genes were examined and the concentrations of urinary hydroxypyrene (1-OHP), 2-naphthol and 8-hydroxydeoxyguanosine (8-OH-dG) were measured. Results : Cigarette smoking was a significant risk factor for lung cancer. The levels of urinary 8-OH-dG were higher in the patients (p<0.001), whereas the urinary 1-OHP and 2-naphthol levels were higher in the controls. The GPX1 codon 198 polymorphism was associated with an increased risk of lung cancer. Individuals carrying the Pro/Leu or Leu/Leu genotype of GPX1 were at a higher risk for lung cancer (adjusted OR=2.29). In addition, these individuals were shown to have high urinary 8-OH-dG concentrations compared to the individuals with the GPX1 Pro/Pro genotype. On the other hand, the polymorphism of the hOGG1 gene did not affect the lung cancer risk and the oxidative DNA damage. Conclusions : These results lead to a conclusion that individuals with the GPX1 Pro/Leu or Leu/Leu genotype would be more susceptible to the lung cancer induced by oxidative stress than those individuals with the Pro/Pro genotype.

Mineral Contents and Physiological Activities of Dried Sea Tangle (Laminaria japonica) Collected from Gijang and Wando in Korea. (기장산과 완도산 건 다시마의 무기성분 및 생리활성 분석)

  • Choi, Jae-Suk;Shin, Su-Hwa;Ha, Yu-Mi;Kim, Yang-Chun;Kim, Tae-Bong;Park, Sun-Mee;Choi, In-Soon;Song, Hyo-Ju;Choi, Young-Ju
    • Journal of Life Science
    • /
    • v.18 no.4
    • /
    • pp.474-481
    • /
    • 2008
  • This research was performed to determine the proximate compositions, mineral contents, alginic acid, antioxidative activities and amino acids of sea tangles collected from Gijang and Wando area. Crude protein and ash contents were higher in Gijang sea tangle, whereas carbohydrate and moisture were higher in Wando in general. Mineral contents of Gijang sea tangle were higher than Wando. Especially, Na and K was the most abundant in both Gijang and Wando sea tangles. Alginic acid content was almost similar in both sea tangles. The major free amino acids were glutamic acid, aspartic acid, alanine, proline and hydroxyproline in both Gijang and Wando sea tangles. Antioxidative activity of methanol extract of sea tangle was measured by using DPPH radical scavenging and SOD-like activity. DPPH radical scavenging and SOD-like activity were about 17% ($40\;{\mu}g/ml$) and 7% ($5\;{\mu}g/ml$) higher, respectively, in Wando sea tangle. When stimulate the macrophages RAW264.7 cells with lipopolysaccharide (LPS), inhibition of NO synthesis of the methanol extract was 11% higher in Wando sea tangle comparing with Gijang samples.

Anti-inflammatory Effect and Antioxidative Activities of Ingredients used in Bibimbab (비빔밥 재료의 항염증 및 항산화 효과)

  • Ko, Yu-Jin;Seol, Hui-Gyeong;Lee, Gyeong-Ran;Jeong, Gye-Im;Ryu, Chung-Ho
    • Journal of Life Science
    • /
    • v.23 no.2
    • /
    • pp.213-221
    • /
    • 2013
  • Bibimbab (mixed rice) is a traditional Korean one-dish meal. This study was carried out to investigate the anti-inflammatory and antioxidative effects of raw and seasoned ingredients used in Bibimbab (Cucurbita moschata P., Platycodon grandiflorum A., Vigna radiata L., Porphyra yezonensis udea, Allium ampeloprasum L., Pterdium aguilinum, Raphanus sativus). Human mast cells (HMC-1) were pretreated with 70% ethanol extracts of Bibimbab and further cultured for an appropriate time after the addition of phorbol 12-myristate 13-acetate (PMA) and calcium ionophore A23187. Cell viability was determined by an MTT assay. None of the ingredients showed cytotoxic effects at a concentration of 1.0 ${\mu}g/ml$. Anti-inflammatory effects were analyzed at 0.01, 0.1, and 1.0 mg/ml concentrations of the raw, seasoned ingredients of PMA. A23187 stimulated HMC-1. Among the various ingredients, seasoned A. ampeloprasum L. extract showed the highest inhibition of TNF-${\alpha}$ and IL-6 secretion (90% and 93%, respectively) at a concentration of 1.0 mg/ml. The R. sativus extract showed the highest inhibition (85%) of IL-8 secretion. DPPH analysis of the antioxidation properties of the ingredients showed that raw and seasoned A. ampeloprasum extracts exhibited the highest DPPH free radical scavenging activity (67.50 and 73.65%, respectively). These results suggest that seasoned ingredients used in Bibimbab have lower anti-inflammatory effects in relation to TNF-${\alpha}$ and IL-6 secretion than raw ingredients in PMA- and A23187-treated HMC-1. In addition, the seasoned ingredients showed a tendency to increase antioxidative activity. Therefore, the ingredients used in Bibimbab have potential as anti-inflammatory and antioxidation agents.

Fructus Sophorae Enhances the Production of Prostaglandin E2 and Tumor Necrosis Factor-α through Activation of MAPKs and PI3K/AKT Signaling Pathways in Murine Macrophages (대식세포에서 MAPKs 및 PI3K/AKT 신호전달계 활성을 통한 괴각 추출물의 prostaglandin E2 및 tumor necrosis factor-α 생성의 촉진)

  • Kang, Young-Soon;Han, Min Ho;Lee, Moon Hee;Hong, Su Hyun;Park, Heungsik;Jung, Jae-Chul;Lee, Jeongrai;Lee, Eun-Woo;Kang, Kyung Hwa;Kim, Cheol Min;Kim, Byung-Woo;Choi, Yung Hyun
    • Journal of Life Science
    • /
    • v.23 no.11
    • /
    • pp.1397-1403
    • /
    • 2013
  • Fructus Sophorae, the dried ripe fruit of Styphnolobium japonicum (L.), is an herbal ingredient used in traditional Oriental medicine. This study was carried out to investigate the effects of Fructus Sophorae extracts (FSE) on immune modulation in a murine RAW 264.7 macrophage model. As immune response parameters, the production of prostaglandin $E_2$ ($PGE_2$) and tumor necrotic $factor-{\alpha}$ ($TNF-{\alpha}$) were evaluated. Our data revealed that FSE increased the macrophage activation and the production of $PGE_2$ and $TNF-{\alpha}$, which was consistently correlated with upregulation of cyclooxygenase-2 (COX-2) and $TNF-{\alpha}$ expression at both transcriptional and translational levels. On comparative cytokine protein array, FSE significantly increased several cytokines, which was associated with phosphorylation of mitogen- activated protein kinases (MAPKs), including extracellular signal-regulated kinase (ERK), p38 MAPK and c-Jun N-terminal kinase (JNK), and Akt in RAW 264.7 cells. However, each inhibitor of these molecules attenuated the FSE-induced $PGE_2$ production. These results indicate that FSE activated macrophages through the activation of MAPKs and phosphatidylinositol-3-kinase (PI3K)/Akt signaling pathways in RAW 264.7 macrophages. These findings suggest that FSE may provide a promising source of an immunoenhancing agent.

Monocentric Derivative Y Chromosome with Duplication of the SRY Gene in an Azoospermic Male (무정자증 불임남성에서 관찰된 SRY 유전자의 중복을 동반한 일동원체성 derivative Y 염색체)

  • Choi, Eun-Young;Lee, Bom-Yi;Park, Ju-Yeon;Lee, Yeon-Woo;Oh, Ah-Rum;Lee, Shin-Young;Kim, Shin-Young;Han, You-Jung;Lee, Mee-Bum;Ryu, Hyun-Mee;Seo, Ju-Tae;Park, So-Yeon
    • Journal of Genetic Medicine
    • /
    • v.7 no.2
    • /
    • pp.160-164
    • /
    • 2010
  • Structural abnormalities of the Y chromosome affect normal testicular differentiation and spermatogenesis. The present case showed a rare monocentric derivative Y chromosome with partial duplication of Yp including the SRY gene and deletion of Yq12 heterochromatin. The karyotype was 46,X,der(Y)(pter${\rightarrow}$q11.23::p11.2${\rightarrow}$pter).ish der(Y)(DYZ3+,DYZ1-,SRY++), confirmed through a FISH study. Even though the patient possessed an abnormal Y chromosome, testicular biopsy showed normal testicular volumes in the proband, with gonadal hormonal levels in the normal range but bilateral varicocele and hypospermatogenesis. We speculate that the abnormal Y chromosome arose from sister chromatids during Y chromosome recombination or intra chromosomal NAHR (non-allelic homologous recombination) during meiosis in the patient's father or in the very early stages of embryogenesis. The derivative Y chromosome might interfere in the meiotic stage of spermatogenesis, leading to the developmental arrest of germ cells. The present case illustrates that the infertility phenotype can have various causes. Also, it emphasizes the importance of accurate and various genetic analyses and could aid in male infertility treatment.

Relationship between the Expression of Sodium Iodide Symporter and the Findings of $^{99m}Tc-MIBI$ Scintimammography in the Primary Breast Cancer (원발성 유방암에서 Sodium Iodide Symporter의 발현과 $^{99m}Tc-MIBI$ 유방스캔의 관계)

  • Seok, Ju-Won;Kim, Seong-Jang;Kwak, Hi-Suk;Lee, Chang-Hun;Kim, In-Ju;Kim, Yong-Ki;Bae, Young-Tae;Kim, Dong-Soo
    • The Korean Journal of Nuclear Medicine
    • /
    • v.36 no.6
    • /
    • pp.325-332
    • /
    • 2002
  • Purpose: Human Na+/I- symporter (hNIS) is known to be expressed in many tissues other than thyroid gland. The breast cancer cells are one of them and the possibility of radioiodine therapy in treatment of the breast cancer may be suggested. We investigated the expression rate of hNIS and the relationship between the expression of hNIS and the finding of 99mTc-MIBI scintimammographv in the breast cancer Materials and Methods: Surgically proved 56 patients with breast cancer were the subjects of this study The expression of hNIS were evaluated by immunohistochemistry and the results were compared to the findings of 99m7c-MIBI scintimammography. Results: Overall expression rate of hNIS was 41.1% in 56 patients. According to the pathologic diagnosis, it was 42.9% in 49 patients with invasive ductal carcinoma and 28.6% in the 7 patients with ductal carcinoma in situ. The expression rate of hNIS in the 41 cases with a focal increased uptake at he breast lesion on 99m7c-MIBI sointimammogram was 31.7%. That in the 15 cases without any abnormal uptake on the scan was significantly higher(65.7%, p<0.05). Conclusion: The expression rate of hNIS in the patients with breast cancer was not so high. The rate was higher in the patients with no increased uptake at the breast lesion on 99m7c-MIBI scintimammography.