• Title/Summary/Keyword: protein detection

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Studies on Host-Virus Interaction of Poxviruses 1. Cytochemical, Autoradiographic and Immunocytological Analysis in Cowpox Virus-FL Cell System (Poxvirus 감염(感染)에 있어서의 Virus-숙주세포(宿主細胞)의 상호관계(相互關係) 1. Cowpox Virus-FL 세포계(細胞系)의 세포화학적(細胞化學的) Autoradiography 및 세포면역학적해석(細胞免疫學的解析))

  • Kim, Uh Ho
    • Korean Journal of Veterinary Research
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    • v.15 no.1
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    • pp.57-67
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    • 1975
  • The poxvirus group is considered to be a typical cytoplasmic inclusion forming virus. Every poxvirus has been reported to produce only one kind of inclusion in the infected tissues. A vague concept that inclusions of poxviruses are eosinophilic or acidophilic has prevailed. Although many papers and theories about the nature of the inclusion have been presented, most of them are not quite convincing on the point of the relations with virus multiplication, and an analysis of papers published showed that there seem to be many discrepancies in the descriptions of the nature of the poxvirus inclusions. Comparative studies on host-virus interaction with cowpox, orf, swinepox and fowlpox viruses which selected from each Group (I-IV) of poxviruses were performed from the morphological and virological standpoints. At first, in cowpox virus-FL cell system, as a comparative model, cytoplasmic inclusion, nucleic acid metabolism by autoradiography and detection of viral antigen by immunofluorescence were studied and obtained the results as follows: 1. The focus-like cytopathic effect (CPE) at early stage developed to entire culture at terminal stage of infection, and also the developing status of CPE was correlated to viral doses for inoculation. Two kinds of cytoplasmic inclusions which named A and B type were easily observed by Giemsa, hematoxylin-eosin (H & E) and May-Greenwald Giemsa (MGG) stainings in the infected cells. The B type inclusions were formed at early stage of infection and the A type inclusions were produced subsequently the B type formation. The B type which common type inclusion in poxviruses was a small compact or aggregate at early stage and developed to a large diffuse body at terminal stage of infection. On the other hand, the A type inclusion which depend upon the kind of virus was appeared as round and discrete shape, and its size and number was increased gradually during the culture period. It was characteristic to form distinct halos around the both types of inclusions in acid fixed, H & E stained preparations of infected cultures. The B type inclusion was always positive in Feulgen reaction and showed as DNA containing body but the A type inclusion was not. 2. In the relationship between inclusion and DNA metabolism of infected cells by the qualitative autoradiography using 3H-thymidine, the appearance of silver grains was coincided with B type inclusion but not with A type inclusion. This showed that the DNA synthesis was proceeded in all B type inclusions except those in the terminal stage with a diffuse form. This suggested that the B type inclusions are only sites of DNA synthesis and this was proceeded after the cell infection independently. The activity of DNA synthesis of the inclusions was nearly the same as that of the nucleic of normal cells and non-inclusion bearing cells. and non-inclusion bearing cells. Regardless of the size of the degree of DNA synthesis of the B type inclusion, inclusion bearing cells all showed remarkable suppression of nuclear DNA synthesis. 3. By the direct fluorescent antibody technique viral antigen in infected cells was detected. The B type inclusions have been proved to contain a great deal of viral antigen, whereas the basic substance of A type inclusion did not show antigenicity except the round edge. It was suggested that the round edge fluorescence might be caused by the glare of cytoplasmic viral antigen which pushed out and concentrated by the A type inclusion development. 4. Hemorrhagic red pock formations on chorioallantoic membrane of embryonated chicken egg had proved the characteristic of used viral strain. 5. By the above studies on the nature of two types of inclusions and the role they play in virus multiplication, it was concluded that the B type inclusion must be the site of the synthesis of viral DNA and protein as well as the site of the virus.

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Allergenicity Change of Soybean Proteins by Thermal Treatment Methods (열처리 방법에 따른 품종별 콩 단백질의 항원성 변화)

  • Seol, Hui-Gyeong;Ko, Yu-Jin;Kim, Eun-Jung;Lee, Gyeong-Lan;Kim, Do-Gyeong;Lee, Jeong-Ok;Ahn, Kang-Mo;Ryu, Chung-Ho
    • Journal of Life Science
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    • v.22 no.4
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    • pp.524-531
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    • 2012
  • Soybean is one of the most common food materials causing food hypersensitivity reactions in Korea. In this study, we have investigated the effect of roasting and fermentation on the allergenicity of soybean. Three kinds of soybean ($Daepung$, $Daewon$, and $Taegwang$) were prepared as raw, roasted, and fermented by $Bacillus$ $subtilis$ GSK 3580, and then their proteins were extracted. The proteins were separated using SDS-PAGE, and the detection of IgE specific to soybean proteins was performed by immunoblotting using 7 sera of soybean allergy patients and non-allergic control individuals. Serum specific IgE to soybean was measured by ELISA. The SDS-PAGE of raw soybean proteins showed various-sized bands ranging from 9 to 76 kDa, which are known as major allergens. In particular, 9, 21, 34, 52, 72, and 76 kDa proteins are known as LTP, Kunits trypsin inhibitor, $Gly$ m Bd 30K, ${\beta}$-subunit, ${\alpha}$-subunit, and ${\alpha}$'-subunit of ${\beta}$-conglycinin, respectively; these are major allergens in soybean. In contrast, only peptides of less than 35 kDa were found in roasted and fermented soybeans. IgE immunoblot analysis of three roasted species of soybeans commonly detected at 38-40 kDa and 10-15 kDa. The protein bands in fermented soybean showed very weak signals or were not detected. In addition, the reactivity of most patients' sera to soybean was decreased after roasting and fermentation. With these results, it may be concluded that the allergenicity of soybeans is reduced by the roasting and fermentation processes. It is supposed that allergenic proteins in soybean were degraded by heat treatment methods and proteolytic enzymes were secreted from fermenting microorganisms.

Effect of the Solvent Fractions of Zingiber officinale Roscoe against Thrombintreated Tumor Invasion in Human Oral Squamous Carcinoma YD-10B Cells (YD-10B 인간구강암세포주에서 생강 유기용매 분획물의 항산화, 트롬빈억제 및 thrombin에 의해 처리된 암 침윤 및 전이 억제 효과)

  • Kim, Eun-Jung;Kim, Jun-Ho
    • Journal of Life Science
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    • v.26 no.11
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    • pp.1289-1297
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    • 2016
  • Oral squamous cell carcinoma (OSCC) is a common malignant tumor in the oral cavity, comprising up to 90% of oral cancer. Oral cancer is characterized by a marked tendency of local invasiveness and is good for early detection and treatment; therefore, it is recognized as a good model for cancer prevention. The present study investigated the antioxidant, thrombin inhibitory, and anti-invasive activities of the solvent fractions of Zingiber officinale Roscoe. Samples were fractionated into hexane, chloroform, ethyl acetate, butanol, and water fractions, and each of these was assayed individually. The water fraction showed the highest extraction yield at 9.79%(w/w). Anti-oxidative activity was analyzed by DPPH assay. Thrombin inhibitory activity was used to analyze thrombin inhibitor assay. Cell viability was detected by the MTS assay. The activity and mRNA expression of MMP-2 and MMP-9 in human oral squamous carcinoma YD-10B cells were examined by zymography and RT-PCR. The antioxidative activities of hexane and water fractions were 92.38% and 92.96%, respectively. In the thrombin inhibitory activity test, water fraction was the highest, with a value of 65.86%. MMP-2/-9 activation was increased in phorbol 12-myristate 13-acetate (PMA)-induced YD-10B cells. MMP-9 activation was increased in thrombin-treated YD-10B cells. In PMA- or thrombin-treated YD-10B cells, the increased mRNA expression and protein activation of MMP-2/-9 were significantly inhibited in the hexane fraction. Therefore, the hexane fraction obtained from a Zingiber officinale Roscoe water extract is a promising therapeutic anti-invasive agent in oral cancer.

DISEASE DIAGNOSED AND DESCRIBED BY NIRS

  • Tsenkova, Roumiana N.
    • Proceedings of the Korean Society of Near Infrared Spectroscopy Conference
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    • 2001.06a
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    • pp.1031-1031
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    • 2001
  • The mammary gland is made up of remarkably sensitive tissue, which has the capability of producing a large volume of secretion, milk, under normal or healthy conditions. When bacteria enter the gland and establish an infection (mastitis), inflammation is initiated accompanied by an influx of white cells from the blood stream, by altered secretory function, and changes in the volume and composition of secretion. Cell numbers in milk are closely associated with inflammation and udder health. These somatic cell counts (SCC) are accepted as the international standard measurement of milk quality in dairy and for mastitis diagnosis. NIR Spectra of unhomogenized composite milk samples from 14 cows (healthy and mastitic), 7days after parturition and during the next 30 days of lactation were measured. Different multivariate analysis techniques were used to diagnose the disease at very early stage and determine how the spectral properties of milk vary with its composition and animal health. PLS model for prediction of somatic cell count (SCC) based on NIR milk spectra was made. The best accuracy of determination for the 1100-2500nm range was found using smoothed absorbance data and 10 PLS factors. The standard error of prediction for independent validation set of samples was 0.382, correlation coefficient 0.854 and the variation coefficient 7.63%. It has been found that SCC determination by NIR milk spectra was indirect and based on the related changes in milk composition. From the spectral changes, we learned that when mastitis occurred, the most significant factors that simultaneously influenced milk spectra were alteration of milk proteins and changes in ionic concentration of milk. It was consistent with the results we obtained further when applied 2DCOS. Two-dimensional correlation analysis of NIR milk spectra was done to assess the changes in milk composition, which occur when somatic cell count (SCC) levels vary. The synchronous correlation map revealed that when SCC increases, protein levels increase while water and lactose levels decrease. Results from the analysis of the asynchronous plot indicated that changes in water and fat absorptions occur before other milk components. In addition, the technique was used to assess the changes in milk during a period when SCC levels do not vary appreciably. Results indicated that milk components are in equilibrium and no appreciable change in a given component was seen with respect to another. This was found in both healthy and mastitic animals. However, milk components were found to vary with SCC content regardless of the range considered. This important finding demonstrates that 2-D correlation analysis may be used to track even subtle changes in milk composition in individual cows. To find out the right threshold for SCC when used for mastitis diagnosis at cow level, classification of milk samples was performed using soft independent modeling of class analogy (SIMCA) and different spectral data pretreatment. Two levels of SCC - 200 000 cells/$m\ell$ and 300 000 cells/$m\ell$, respectively, were set up and compared as thresholds to discriminate between healthy and mastitic cows. The best detection accuracy was found with 200 000 cells/$m\ell$ as threshold for mastitis and smoothed absorbance data: - 98% of the milk samples in the calibration set and 87% of the samples in the independent test set were correctly classified. When the spectral information was studied it was found that the successful mastitis diagnosis was based on reviling the spectral changes related to the corresponding changes in milk composition. NIRS combined with different ways of spectral data ruining can provide faster and nondestructive alternative to current methods for mastitis diagnosis and a new inside into disease understanding at molecular level.

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Shelf-life of Prepacked Kimbab and Sandwiches Marketed in Convenience Stores at Refrigerated Condition (편의점에서 판매되는 김밥 및 샌드위치의 냉장조건에서의 유통기한)

  • Koo, Min-Seon;Kim, Yoon-Sook;Shin, Dong-Bin;Oh, Se-Wook;Chun, Hyang-Sook
    • Journal of Food Hygiene and Safety
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    • v.22 no.4
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    • pp.323-331
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    • 2007
  • This study was designed to estimate self-life of Kimbab and sandwiches marketed in convenience store. While the 12 different type of Kimbab (n=6) and sandwiches (n=6) were kept at $10^{\circ}C$ for 72 hours, quality changes including volatile basic nitrogen, aerobic plate count, pathogens detection and sensorial property was monitored, and effective quality indicators were selected. Volatile basic nitrogen, indicator for protein deterioration was slightly increased during storage periods in all samples. E. coli, Staphylococcus aureus, Salmonella spp. and Vibrio parahaemolyticus were not detected from any of samples. Change of aerobic plate count of Kimbab and sandwiches were increased moderately but increased dramatically after 48 hours of storage. Overall acceptability were maintained over 5, purchasing power limit, for 40 hours in 4 general Kimbab, 48 hours in 2 samgak Kimbab and 42 hours in 2 sandwiches. Shelf-life of each item was calculated from regression equation between reference limit from effective quality indicators, aerobic plate count and sensory property, and storage period. Estimated shelf-lives of general Kimbab were $15{\sim}33$ hours, samgak Kimbab were 32 hours and sandwiches were $27{\sim}30$ hours at $10^{\circ}C$ refrigerated condition.

Detection of Human Papillomavirus and Expression of p53, c-erbB-2 Protein in Inverted Papilloma of the Nasal Cavity and Paranasal Sinuses (비강 및 부비동의 반전성 유두종에서 인유두종바이러스검출과 p53및 c-erbB-2의 발현)

  • Cho Jae-Shik;Baik Joon;Lim Sang-Chul;Cho Yeon;Yoon Je-Hwan;Seo Duk-Jung;Park Chang-Soo
    • Korean Journal of Head & Neck Oncology
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    • v.17 no.2
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    • pp.162-168
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    • 2001
  • Background: Inverted papilloma(IP) of the nasal cavity and paranasal sinuses is a benign neoplastic condition that can be associated with squamous cell carcinoma (SCC). Several studies have indicated an etiologic role for viruses in the development of inverted papilloma. And it is necessary to find out the significance of a biologic markers such as p53, c-erbB-2 to predict the malignant potential. The purposes of this study are to detect HPV in inverted papilloma of the nasal cavity and paranasal sinus, to examine role of HPV as an etiological agent, to examine the relationship between HPV subtype and malignant transformation of inverted papilloma, and to investigate the relation between expression rate of p53, c-erbB-2 and HPV in recurrent or malignant transformation cases. Material and Methods: Thirty two cases of inverted papilloma(IP) in the nasal cavity and paranasal sinuses were reviewed and classified into 3 groups; simple IP, IP with dysplasia group, IP with squamous cell carcinoma group. Paraffin embedded achival tissue was used in this study. The HPV was detected by in situ hybridzation (ISH) using HPV type 6/11, 16/18, 31/33/35 DNA probes. Expression of p53 and c-erbB-2 was examined by immunohistochemical staining. Results: 1) The HPV was detected in 6(19%) out of 32 cases. 2) The HPV 6/11 was dectected in 4 out of 21 cases of simple IP, HPV 16/18 in 1, HPV 31/33/35 in lout of 8 cases of IP with dysplasia respectively. 3) The positive expression of p53 was 13 cases out of 32 cases; 2 out of 21 cases of simple IP, all of 8 cases of IP with dysplasia and 3 cases of IP with squamous cell carcinoma 4) The positive expression of c-erbB-2 was in 24 out of 32 cases; 16 out of 21 cases of simple IP, 6 out of 8 cases of IP with dysplasia, 2 out of 3 cases of IP with squamous cell ca. 5) The recurrence of IP occurred in lout of 6 cases of positive for HPV, in 4 out of 26 cases negative for HPV. 6) The recurrence of IP occurred only in positive cases for p53. 7) The recurrence of IP occurred in 4(17%) out of 24 cases positive for c-erbB-2, in 1(13%) out of 8 cases negative for c-erbB-2. Conclusion: The p53 expression was associated with Inverted papillomas exhibiting evidence of malignant transformation. Also, there was a correlation between the p53 expression and recurrence.

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Effects of Glufosinate-Ammonium to Earthworms, Soil Microorganisms and Crops (제초제 glufosinate-ammonium의 지렁이 및 토양 미생물과 작물에 미치는 영향)

  • Kim, Yong-Seog;Jeon, Yong-Bae;Choi, Hae-Jin;Kim, Song-Mun;Kim, Sung-Min
    • The Korean Journal of Pesticide Science
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    • v.10 no.2
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    • pp.76-83
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    • 2006
  • In order to investigate the impacts of non-selective herbicide, glufosinate-ammonium (ammonium 4-[hydroxy(methyl)phosphinoyl] -DL-homoalaninate, GLA) to the non-target organisms, earthworm was exposed to GLA in the field soil for a month, and microbial populations in the soil were investigated after application of GLA. Simultaneously, the residues of GLA and its metabolite, 3-MPP were analyzed in the same soil. Meanwhile, to elucidate the influence of GLA to the growth of non-target crops incase of inter-furrow application, the amounts of carotenoid, chlorophyll, amino acid, proteins and sugars in the leaves of potato and chinese cabbage grown in the same field were investigated. In result, the dead earthworm was not observed during the test period, and the increasing rates of bodyweight were $9.410{\sim}11.603%$ in GLA-treated plots and 5.645% in GLA-untreated plots. The populations of fungi, bacteria and actinomycetes in the GLA-treated soils were $6.2{\times}10^4$, $1.5{\times}10^6$ and $5.7{\times}10^4$, respectively. They maintained relatively similar levels to the control which were $3.7{\times}10^4$, $3.7{\times}10^5$ and $3.7{\times}10^4$, respectively. In residue analysis, the limit of detection of GLA was 0.02 mg $kg^{-1}$, that of 3-MPP was the same level, and the half-life of GLA was 15 days in sandy clay loam soil. This result indicates that GLA was degraded very quickly in field soil. On the other hand, the amounts of physiological, biochemical components such as carotenoid, amino acid, chlorophyll, protein and sugar were ranged from 90.0 to 104.3% in potato and from 99.0 to 112.7% in chinese cabbage. Comparing with hand-weeded plots, it is indicated that GLA had not affected to the growth of non-target crops when applied at inter-furrow in crops-growing field.

Effects of Reactivation of Latent Epstein-Barr Virus Using Polymerase Chain Reaction on Acute Hepatitis A in Children (중합효소연쇄반응으로 확인된 엡스타인-바바이러스 재활성화가 소아 급성 A형 간염에 미치는 영향)

  • Baek, Seung-Hyon;Kim, Sang-Yong;Koh, Hong
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.14 no.1
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    • pp.59-66
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    • 2011
  • Purpose: We previously reported that concurrent reactivation of latent Epstein-Barr virus (EBV) in children with hepatitis A virus (HAV) infection is common and EBV reactivation with HAV infection adversely affects the clinical features of hepatitis. However, the incidence of concurrent reactivation was not accurate because the detection of EBV reactivation was based on serologic methods. Therefore, we studied the effects of polymerase chain reaction (PCR)-proven EBV reactivation, thus a more precise concurrence, on acute HAV infection in children. Methods: PCR were conducted in 34 patients, who had enrolled previous study and diagnosed with acute HAV infection between January 2008 and June 2010. Their medical records were reviewed. Results: Among 34 patients with acute HAV infection, 12 patients (35.3%) had EBV reactivation which was proven using serologic and molecular biologic techniques. There were significant differences in the peak levels of AST and ALT between the reactivated and non-reactivated groups (p=0.001 and p<0.001, respectively). The duration of full recovery from hepatitis was more prolonged in the reactivated group (p<0.001). Clinical parameters, such as serum protein (p<0.001) and albumin concentrations (p<0.001), atypical lymphocyte count (p=0.001), prothrombin time-international normalized ratio (PT-INR, p<0.001), and splenomegaly (p<0.001), showed significant differences. The clinical features in the reactivated sub-group >10 years of age revealed more liver dysfunction compared to the non-reactivated sub-group. A comparison with a previous study was performed. Conclusion: PCR-proven reactivation of latent EBV in children with HAV infection is common and EBV reactivation with HAV infection adversely affects the clinical features of hepatitis, especially in older children.

Composition of Amino Acids, Minerals, and Heavy Metals in Differently Cooked Laver (Porphyra tenera) (김(Porphyra tenera)의 조리방법에 따른 아미노산, 무기질, 중금속 함량 분석)

  • Hwang, Eun-Sun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.8
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    • pp.1270-1276
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    • 2013
  • Laver, a red algae belonging to the genus Porphyra tenera, is one of the most widely consumed edible seaweeds in Korea, China, and Japan. Lavers are usually consumed in dried, roasted, and seasoned forms to improve their palatability. We evaluated the composition of amino acids, minerals, and trace heavy metals in these three differently cooked forms of laver. The moisture and ash contents of three differently cooked lavers ranged from 1.49~9.69% and 6.07~10.31%, respectively. The crude protein and lipid content ranged from 17.24~36.88% and 0.52~42.42%, respectively. Dried laver was found to be a good source of amino acids such as taurine, alanine, and glutamic acid (871.10 mg, 833.53 mg, and 719.77 mg per 100 g dry weight, respectively). Laver was a good source of macro and micro minerals such as K, Ca, Mg, Na, P, I, and Fe, although laver more extensively cooked (roasted and seasoned) contained less minerals compared to the dried form. Mercury levels in the three differently cooked forms of laver were all less than 100 ng/g dry weight (the limit of detection with our methodology). The levels of arsenic were the most abundant elements in the differently cooked laver. There was a clear variation, depending on the cooking process, in terms of amino acid, mineral, and trace metal contents of laver.

Low Frequency Noise Induces Stress Responses in the Rat (흰쥐에서 저주파소음에 의한 스트레스 반응)

  • Choi, Woong-Ki;Lee, Kyu-Sop;Joung, Hye-Young;Lee, Young-Chang;Sohn, Jin-Hun;Lee, Bae-Hwan;Pyun, Kwang-Ho;Shim, In-Sop
    • Science of Emotion and Sensibility
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    • v.10 no.3
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    • pp.411-418
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    • 2007
  • Exposure to low frequency noise(LFN) can lead to vibroacoustic diseases(VADs), which include a systemic disease with lesions in a broad spectrum of organs and a psychiatric condition. It is known that VAD is an established risk factor for the development of many psychological conditions in humans and rodents, including major depression and anxiety disorder. The present study investigated the effects of LFN on neuronal stress responses in the rat brain. The neuronal expression of the proto-oncogene c-fos in the paraventricular nucleus(PVN) of the hypothalamus and tyrosine hydroxylase(TH) in the LC was observed. The immunocytochemical detection of the Fos protein and TH has been used as a marker of neuronal activation in response to stress. In addition, corticosterone concentration was evaluated by using an enzyme-linked immunosorbent assay(ELISA). The LFN groups were exposed to 32.5Hz and 125Hz of noise(4hr/day for 2days). The numbers of c-fos and TH-immunoreactive cells in the PVN and LC were significantly increased in the LFN groups(32.5Hz and 125Hz) compared to the normal group. Corticosterone concentration in plasma was also increased in LFN groups. The present results demonstrated that exposure with LFN produced a pronounced increase in expression of c-Fos and TH in stress-relevant brain areas. These results suggest that the neural characteristics involved in LFN are similar to those activated by typical processive stressors. These results also suggest that the central and peripheral activations by LFN may be related to LFN-related negative behavioral dysfunctions such as VADs.

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