This study was conducted for quantitative microbial risk assessment (QMRA) of Clostridium perfringens with consumption on ham and sausage products in Korea, according to Codex guidelines. Frame-work model as product-retail-consumption pathway composed with initial contamination level, the time and temperature in distributions, and consumption data sets for ham and sausage products and also used the published predictive growth and dose-response models for Cl. perfringens. The simulation model and formulas with Microsoft@ Excel spreadsheet program using these data sets was developed and simulated with @RISK. The probability of foodborne disease by Cl. perfringens with consumption of the ham and sausage products per person per day was estimated as $3.97{\times}10^{-11}{\pm}1.80{\times}10^{-9}$. There were also noted that limitations in this study and suggestion for development of QMRA in the future in Korea.
Amyloid-${\beta}$ peptide ($A{\beta}$), generated by proteolytic cleavage of the amyloid precursor protein (APP), plays a pivotal role in the pathogenesis of Alzheimer's disease (AD). The key step in the generation of $A{\beta}$ is cleavage of APP by beta-site APP-cleaving enzyme 1 (BACE1). Levels of BACE1 are increased in vulnerable regions of the AD brain, but the underlying mechanism is unknown. In the present study, we reported the effects of ferrous ions at subtoxic concentrations on the mRNA levels of BACE1 and a-disintegrin-and-metalloproteinase 10 (ADAM10) in PC12 cells and the cell responses to ferrous ions. The cell survival in PC12 cells significantly decreased with 0 to 0.3 mM $FeCl_2$, with 0.6 mM $FeCl_2$ treatment resulting in significant reductions by about 75%. 4,6-diamidino-2-phenylindole (DAPI) staining showed that the nuclei appeared fragmented in 0.2 and 0.3 mM $FeCl_2$. APP-${\alpha}$-carboxyl terminal fragment (APP-${\alpha}$-CTF) associations with ADAM10 and APP-${\beta}$-CTF with BACE1 were increased. Levels of ADAM10 and BACE1 mRNA increased in response to the concentrations of 0.25 mM, respectively. In addition, p-ERK and p-Bad (S112, S155) expressions were increased, suggesting that APP-CTF formation is related to ADAM10/ BACE1 expression. Levels of Bcl-2 protein were increased, but significant changes were not observed in the expression of Bax. These data suggest that ion-induced enhanced expression of AMDA10/BACE1 could be one of the causes for APP-${\alpha}/{\beta}$-CTF activation.
Electron microscopic studies were made to investigate changes in the fine structure of the liver, kidney and gills of Carassius carassius L. following exposure to 1 and 2.5 ppm of $HgCl_2$. The following results were obtained: 1. In the mercury-treated liver cells, an increase in the number of lysosomes were noticed. These lysosomes appeared to be of two types; round ones containing some crystalline structures and others with phagocytosed glycogen granules and mitochondria. Also observed were mitochondrial swelling where the matrix appeared less electrondense, and segregation of the nucleoli in the nucleus. 2. In the kidney, mercury treatment resulted in thickening of the basement membrane of the glomerulus, and appearance of vacuoles and cytoplasmic bodies in the proximal convoluted tubule. The vacuoles seemed to be formed from mitochondria. Nuclear shrinkage was also noticed at 2.5 ppm of $HgCl_2$. 3. Many large and small lysosomes appeared in response to mercury in the epithelial cells of the gill lamella. Also the lamellar membrane became fuzzy in appearance. 4. It can be concluded from these results that mercury-induced changes in the fine structure are associated with activation of detoxication processes and impairment of energy metabolism.
Kim, Il-Sup;Shin, Sun-Young;Kim, Young-Saeng;Kim, Hyun-Young;Yoon, Ho-Sung
Molecules and Cells
/
v.28
no.5
/
pp.479-487
/
2009
Glutathione reductase (GR) is an enzyme that recycles a key cellular antioxidant molecule glutathione (GSH) from its oxidized form (GSSG) thus maintaining cellular redox homeostasis. A recombinant plasmid to overexpress a GR of Brassica rapa subsp. pekinensis (BrGR) in E. coli BL21 (DE3) was constructed using an expression vector pKM260. Expression of the introduced gene was confirmed by semi-quantitative RT-PCR, immunoblotting and enzyme assays. Purification of the BrGR protein was performed by IMAC method and indicated that the BrGR was a dimmer. The BrGR required NADPH as a cofactor and specific activity was approximately 458 U. The BrGR-expressing E. coli cells showed increased GR activity and tolerance to $H_2O_2$, menadione, and heavy metal ($CdCl_2$, $ZnCl_2$ and $AlCl_2$)-mediated growth inhibition. The ectopic expression of BrGR provoked the co-regulation of a variety of antioxidant enzymes including catalase, superoxide dismutase, glutathione peroxidase, and glucose-6-phosphate dehydrogenase. Consequently, the transformed cells showed decreased hydroperoxide levels when exposed to stressful conditions. A proteomic analysis demonstrated the higher level of induction of proteins involved in glycolysis, detoxification/oxidative stress response, protein folding, transport/binding proteins, cell envelope/porins, and protein translation and modification when exposed to $H_2O_2$ stress. Taken together, these results indicate that the plant GR protein is functional in a cooperative way in the E. coli system to protect cells against oxidative stress.
Ahmed, Rania S;Embaby, Amira M;Hassan, Mostafa;Soliman, Nadia A;Abdel-Fattah, Yasser R
Microbiology and Biotechnology Letters
/
v.49
no.2
/
pp.181-191
/
2021
The present study addresses isolation, optimization, partial purification, and characterization of a haloalkaline serine protease from a newly isolated haloarchaeal strain isolated from Wadi El Natrun in Egypt. We expected that a two-step sequential statistical approach (one variable at a time, followed by response surface methodology) might maximize the production of the haloalkaline serine protease. The enzyme was partially purified using Hiprep 16/60 sephacryl S-100 HR gel filtration column. Molecular identification revealed the newly isolated haloarchaeon to be Natrialba hulunbeirensis strain WNHS14. Among several tested physicochemical determinants, casamino acids, KCl, and NaCl showed the most significant effects on enzyme production as determined from results of the One-Variable-At-A-time (OVAT) study. The BoxBehnken design localized the optimal levels of the three key determinants; casamino acids, KCl, and NaCl to be 0.5% (w/v), 0.02% (w/v), and 15% (w/v), respectively, obtaining 62.9 U/ml as the maximal amount of protease produced after treatment at 40℃, and pH 9 for 9 days with 6-fold enhancement in yield. The enzyme was partially purified after size exclusion chromatography with specific activity, purification fold, and yield of 1282.63 U/mg, 8.9, and 23%, respectively. The enzyme showed its maximal activity at pH, temperature, and NaCl concentration optima of 10, 75℃, and 2 M, respectively. Phenylmethylsulfonyl fluoride (PMSF, 5 mM) completely inhibited enzyme activity.
The barley grown in pot-soil was treated with the NaCl solution of -20 bar in osmotic potential for 10 days, varying the time of treatment: from 20th day before heading; from 10th day before heading and the time of heading. The greatest injury was observed in the case of salt stress at heading or at 10th day before heading: Culm length decreased by 87% : the number of spikes per plant by 82% ; the number of grain per spike by 92% : 1, 000-grain weight by 94% ; yield per pot by 75%. The results also greatly varied depending upon the cultivars and the time of salt stress. Salt stress at the time of heading or at 10th day before heading remarkably decreased yield and yield components. And in terms of grain yield the salt resistance was high in the order of Baegdong, Albori, Hyangmaeg, Olbori and Durubori.
Exposure of the root surface due to gingival recession after periodontal surgery, elicit pain response when exposed to mechanical, heat, chemical or osmotic irritation. Especially patients treated with periodontal surgery, show high frequency. There have been reports that the 1 out of 7 patients complains of dentinal hypersensitivity. There have been many studies on the clinical effects of various materials on the treatment of dentinal hypersensitivity. The purposes of this study were to evaluate the effect of sodium chloride and potassium oxalate and to observe the relationship between the dentinal hypersensitivity and surface characteristics such as dentinal tubule size and number. This study included 20 teeth which were scheduled for extraction and had no pulpal disease. These teeth were divided into Root planing group, EDTA group, NaCl group and Oxalate group. Dentinal hypersensitivity is measured by tactile, pressured air and cold water using NRS (Numerical Rating Scales). Teeth were extracted under local anesthesia and each specimen was sectioned to a size about 3 X 5 mm and was examined under the scanning electron microscope (X2,000) The results were as follows, 1. The EDTA group exhibited significantly increased dentinal hypersensitivity comparing with the other groups. 2. The NaCl and Oxalate groups showed significantly reduced dentinal hypersensitivity comparing with the EDTA group. 3. As a method for dentinal hypersensitivity measurement, it was presumed thet tactile sensitivity test was not sensitive method but air blast test and cold water test were adequate method. 4. In a SEM study, the root planing group exhibited amorphous smear layer and showed no dentinal tubule orifice, but the EDTA group showed the large number of dentinal tubules. On the other hand, the NaCl and Oxalate groups did not show exposed dentinal tubules. The NaCl group showed more rough root surface than the EDTA group, and the Oxalate group showed many participates to be presumed as calcium oxalate particle. As the results from this study, root planing couldn't expose the dentinal tubule and NaCl and potassium oxalate occluded exposed dentinal tubule effectively. Dentinal hypersensitivity has close relationship with the exposure of dentinal tubules, especially with it's size and number.
Kim, Nam-Song;Lee, Jae-Hyung;Koh, Dai-Ha;Ki, No-Suk;Hwang, In-Dam
Journal of Preventive Medicine and Public Health
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v.24
no.3
s.35
/
pp.287-304
/
1991
Tolerance to several toxic effects of cadmium, including lethality has been shown following pretreatment with cadmium and zinc. This study was designed to determine if tolerance also develops to Cd-induced hepatotoxicityandrenaltoxicity. Three groups of rats (A, B, C), each consisting of 16 rats, were studied and each group was divided into four subgroups (1, 2, 3, 4), 4 rats for each subgroup. Rats were subcutaneously pretreated with saline (A), $CdCl_2$ (0.5 mg/kg, B), and $ZnCl_2$ (13.0 mg/kg, C) during time periods of $1{\sim}6$ weeks. At the end of the period, rats were challenged with $CdCl_2$ (3.0, 6.0 and 9.0 mg/kg, ip). After giving the challenge dose, cadmium and metallothionein (MT) concentrations were determined and also observed the histologic change in liver and kidney. The concentration of cadmium in liver and kidney increased dose-dependently to the challenge dosage. These da indicate the kidney is a major target organ of chronic cadmium poisoning, and suggest that cadmium induced hepatic injury, via release of Cd-MT, may play an important role in the nephrotoxicity observed in response to long-term exposure to cadmium. In addition, histologic examination of group $A_2,\;A_3\;and\;A_4$ revealed moderate to severe cadmium toxicity, evidenced by infiltration of inflammatory cells, cell swelling, pyknosis, enlarged sinusoids and necrosis in liver, and tubule cell necrosis and degeneration in kidney. However, MT concentrations in liver and kidney were increased by the pretreatment of $CdCl_2$ and $ZnCl_2$, and their morphological findings were not significantly changed, comparing with control group. Higher MT concentration in liver and kidney observed in the pretreated groups constitutes a plausible explanation of the protective effects of pretreatment against the cadmium toxicity after challenge dosing.
Park, Hea-Woon;Son, Jo-Young;Cho, Yun-Woo;Hwang, Se-Jin;Kim, Su-Jeong;Ahn, Sang-Ho;Jang, Sung-Ho;Jung, Yong-Jae
The Journal of Korean Physical Therapy
/
v.22
no.3
/
pp.99-105
/
2010
Purpose: The purpose of this study was to evaluate the development of pain behavior and the expression of CCL2/CCR2 and CX3CL1/CX3CR1 at above and below the level of hemisection of the spinal cord in a rat model. Methods: Spinal cords of adult female Sprague-Dawley rats (n= 16, 200~250 g, 6~8 weeks old) were hemisected at T13 on the right side to develop the spinal hemisection injury model. We compared behavioral responses of the hemisection and of a sham surgery group. Behavioral tests for motor function (by the BBB locomotor scale), and for pain response for mechanical and cold allodynia were assessed postoperatively (PO) for 21 days. Expression of mRNA for chemokines and their receptors (CCL2/CCR2 and CX3CL1/CX3CR1) below and above the level of the spinal cord dissection were examined by RT-PCR. Results: We observed gradual motor improvement and the development of mechanical and cold allodynia on the ipsilateral hindpaw after spinal hemisection injury. We also found upregulation of mRNA expression of CCL2/CCR2 both above and below the level of spinal cord dissection but CX3CL1/CX3CR1 mRNA expression. Conclusion: Upregulation of CCL2/CCR2 is associated with neuropathic pain after spinal hemisection injury. CCL2/CCR2 may play an important role in the development of neuropathic pain after SCI as well as of peripheral neuropathic pain. These findings may improve understanding of the pathophysiological mechanism of neuropathic pain after SCI.
Kim, Tae-Kyung;Ku, Su-Kyung;Lee, Hye-Jin;Lee, Cheol-Won;Kim, Young-Boong;Jeon, Ki-Hong;Choi, Yun-Sang
Korean journal of food and cookery science
/
v.32
no.6
/
pp.716-723
/
2016
Purpose: This study was conducted to improve the quality characteristics of pork intestine through different pretreatment processes. Methods: We washed pork intestine by both physical (tap water, UV, and sonication) and chemical methods (alcohol, acetic acid, flour and NaCl) as pretreatment process. The physicochemical (pH, color, volatile basic nitrogen (VBN), and 2-thiobarbituric acid reactive substances (TBARS)) and microbial properties of pre-treated pork small intestine were evaluated. Results: The nature of the pretreatment method influenced the pH value of pork small intestine. The acetic acid treatment resulted in the lowest pH value. In physical method, the color value and the number of microorganism were significantly affected by sonication as compared to other treatments. TBARS value of pork small intestine after all the treatments was lower than the control. However, VBN exhibited no significant differences in its value irrespective of the nature of treatment. Appearance and control exhibited lowest value in response to sonication treatment. However, off-flavor and overall acceptability were higher in sonication treatment than other treatments. In chemical method involving NaCl and flour treatments, lightness and redness were lower than other treatments. Lowest VBN and TBARS values were noted in alcohol and acetic acid treatmentsand no growth of E. coli and coliform bacteria was observed. The other treatments resulted in lower values of VBN, TBARS, and microbial counts than the control. Appearance and color value after alcohol, acetic acid, and flour treatment were lower than the control and NaCl treatment. Off-flavor and overall acceptability of by-product after alcohol, flour, and NaCl treatments were higher than the control and acetic acid treatment. Conclusion: Overall, we present NaCl treatment and sonication treatment in the form of a combination pretreatment method as the optimal condition for processing pork intestine.
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