• Title/Summary/Keyword: Cell cultures

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The Purification and Immunogenicity of TB-14 Recombinant Protein of Mycobacterium tuberculosis (결핵균 특이 TB-14 재조합 단백질의 분리 및 세포성 면역반응에 미치는 영향)

  • Song, Ho-Yeon;Kim, Young-Hee;Kim, Chang-Hwan;Min, Young-Ki;Kim, Dae-Joong;Ko, Kwang-Kjune
    • Tuberculosis and Respiratory Diseases
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    • v.61 no.3
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    • pp.239-247
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    • 2006
  • Background: Culture filtrate proteins secreted by mycobacteria are thought to play an important role in inducing protective immunity and to develop new methods for diagnosing tuberculosis. Methods: A culture filtrate protein of M. avium that was strongly reactive with goat antiserum against M. intracellulare was constructed. Its homologous protein (TB-14) in M. tuberculosis was cloned, expressed and purified. The inductions of IFN-${\gamma}$ stimulated with $10{\mu}g$ of TB-14 recombinant protein and $10{\mu}g$ PPD were estimated by using whole bloods from seven PPD (-) subjects, seven PPD (+) healthy volunteers and nine tuberculosis patients. Results: M. avium culture filtrate protein was confirmed as a hypothetical protein that was termed contig 116. A novel 14-kDa recombinant protein (TB-14) of M. tuberculosis was composed of 148 amino acids, including 30 amino acids of the signal peptide, and it showed 78% homology with M. avium. In the PPD (+) healthy volunteers, recombinant TB-14 protein strongly induced the secretion of IFN-${\gamma}$ in whole blood cultures. Conclusion: These results suggest that TB-14 recombinant protein might play an important role in inducing cell-mediated immunity against tuberculosis. Furthermore, TB-14 protein antigen and its antiserum will be available for the development of new diagnostic tools for tuberculosis.

The Hepatotprotective and Antioxidative Effects of Onion (Allium cepa) Extracts in Rat Hepatocyte Primary Culture (양파(Allium cepa) 추출물의 간보호 및 항산화 효과)

  • Rhim, Tae-Jin;Lim, Sang-Cheol
    • Proceedings of the Plant Resources Society of Korea Conference
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    • v.18 no.1
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    • pp.52-60
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    • 2005
  • The objectives of present study were to investigate the hepatoprotective and antioxidative effects of onion extracts. Primary cultures of rat hepatocytes were incubated with 1.5 mM tert-butyl hydroperoxide(t-BHP), potent oxidizing agent for liver injury for 1 hr in the presence or absence of various concentrations (0, 0.01, 0.05, 0.1 or 0.3 mg/ml) of onion extract. Cytotoxicity and cell viability were determined by measuring glutamic oxaloacetic transaminase(GOT) activity, lactate dehydrogenase(LDH) activity and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide(MTT) value. Lipid peroxidation was evaluated using thiobarbituric acid reactive substances(TBARS) assay. Effects on antioxidant system were determined by measuring catalase, glutathione peroxidase(GSH-Px), glutathione reductase(GSH-Rd) activities as well as DNA strand breaking assay. Incubation with t-BHP alone increased GOT and LDH activities and TBARS concentration but decreased MTT reduction. Onion extracts at the concentration of 0.05 mg/ml began to decrease GOT and LDH activities induced by 1.5 mM t-BHP. Decreased MTT reduction began to be increased by onion extract at the concentration of 0.01 mg/ml. Onion extracts at the concentration of 0.01 mg/ml began to decrease TBARS concentration induced by t-BHP. Taken together, onion extracts prevented t-BHP-induced hepatocyte injury and lipid peroxidation. Catalase, GSH-Px and GSH-Rd activities of hepatocytes were significantly decreased by 1.5 mM t-BHP for 1 hr incubation. Onion extracts, on the other hand, at the concentration of 0.1 mg/ml began to prevent t-BHP-induced decrease in catalase, GSH-Px and GSH-Rd activities. Onion extracts prevented hydroxyl radical-induced single-strand breakage in dose-dependent manner when plasmid DNA was incubated with various concentrations of onion extracts in the presence of Fenton regents producing hydroxyl radical. These results demonstrate that onion extracts suppressed t-BHP-induced cytoctoxicity, decreased viability and lipid peroxidation and increased GSH-Px, GSH-Rd and catalase activities. Thus hepatoprotective and antioxidant effects of onion extract seem to be due to, at least in part, the increase in antioxidant enzyme activities as well as prevention from hydroxyl radical-induced oxidation, followed by inhibition in lipid peroxidation.

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THE ASPECT OF PROLIFERATION AND BONE NODULE FORMATION IN OSTEOBLAST-LIKE CELLS DERIVED FROM FETAL RAT CALVARIA IN VITRO (백서 태자 두 개관에서 유래된 조골세포의 증식 및 골결절 형성양상)

  • Kim, Shi-Hyeong;Nam, Soon-Hyeun;Shin, Hong-In
    • Journal of the korean academy of Pediatric Dentistry
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    • v.24 no.1
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    • pp.1-17
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    • 1997
  • The purpose of this study was to investigate the aspects of proliferation and bone nodule formation of osteogenic precursor cells. To determine the effects of ascorbic acid and dexamethasone upon capacity of osteoblast proliferation and bone nodule formation, cells were maintained in the presence of one or some of these additives for up to 30 days. Group I culture was maintained in standard medium(DMEM plus 10% plus antibiotics), group II was maintained in supplemented medium containing dexamethasone, group III was maintained in supplemented medium containing ascorbic acid and sodium-${\beta}$-glycerophosphate, and group IV was maintained in supplemented containing ascorbic acid, sodium-${\beta}$-glycerophosphate and dexamethasone. Morphology of bone nodules was observed with light microscope and electron microscope. The results were as follows: ${\bullet}$ Proliferation capacity of osteoblasts was not affected by single use of dexamethasone, but it was chiefly affected by ascorbic acid. ${\bullet}$ Cellular morphology was fibroblastic appearance initially, but, it was gradually changed to polygonal shape accompanied by confluency stage. ${\bullet}$ Pluripotent mesenchymal cells existed during primary culture, they were differentiated to adipocyte, chondrocyte, osteocyte according to culture condition. ${\bullet}$ Dexamethasone increased bone nodule formation under the condition that the culture was maintained with supplemented medium ascorbic acid and sodium-${\beta}$-glycerophosphate. ${\bullet}$ when the cultures were stained with alizarin red, the group supplemented with dexamethasone, ascorbic acid and sodium-${\beta}$-glycerophosphate showed the marked increase of bone nodule formation, but the group supplemented with ascorbic acid and sodium-${\beta}$-glycerophosphate revealed only small amounts of bone nodules. And the groups cultured without ascorbic acid showed no observed any of bone-like mass independent of dexamethasone addition.

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Etiology and Clinical Manifestations of Fever in Infants Younger than 3 Months Old: A Single Institution Study, 2008-2010 (3개월 미만 영아의 발열에 대한 원인 및 임상양상에 대한 단일기관 연구; 2008-2010)

  • Seok, Joon Young;Kang, Ji Eun;Cho, Eun Young;Choi, Eun Hwa;Lee, Hoan Jong
    • Pediatric Infection and Vaccine
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    • v.19 no.3
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    • pp.121-130
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    • 2012
  • Purpose : The purpose of this study is to investigate clinical features and causative organisms in febrile infants younger than three months, to help identification of high risk patients for serious bacterial infection (SBI). Methods : A total of 313 febrile infants younger than three months, who had visited Seoul National University Children's Hospital from January 2008 to December 2010 were included. Clinical features, laboratory findings, causative organisms, and risk factors of SBI were analyzed by retrospective chart review. Causative bacterial or viral pathogens were identified by gram stain and cultures, rapid antigen tests, or the polymerase chain reaction from clinically reliable sources. Results : Among 313 infants, etiologic organisms were identified in 127 cases (40.6%). Among 39 cases of bacterial infections, Escherichia coli (66.7%) and Streptococcus agalactiae (12.8%) were common. Enterovirus (33.7%), respiratory syncytial virus (19.8%), and rhinovirus (18.8%) were frequently detected in 88 cases of viral infection. Patients with SBI (39 cases) showed significantly higher values of the white blood cell count ($14,473{\pm}6,824/mm^3$ vs. $11,254{\pm}5,775/mm^3$, P=0.002) and the C-reactive protein ($6.32{\pm}8.51mg/L$ vs. $1.28{\pm}2.35mg/L$, P<0.001) than those without SBI (274 cases). The clinical risk factors for SBI were the male (OR 3.7, 95% CI 1.5-8.9), the presence of neurologic symptoms (OR 4.8, 95% CI 1.4-16.8), and the absence of family members with respiratory symptoms (OR 3.6, 95% CI 1.2-11.3). Conclusion : This study identified common pathogens and risk factors for SBI in febrile infants younger than three months. These findings may be useful to guide management of febrile young infants.

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Diagnostic Values of pH of Vaginal Discharge, Amine Test, and Microscopic Examination of Infectious Vaginitis (감염성 질염에 대한 질내 산도측정, Amine 검사 및 현미경 검사의 진단적 가치)

  • Choi, Jae-Dong;Kim, Gi-Duk;Koh, Min-Whan;Lee, Tae-Hyung;Chung, Wun-Yong
    • Journal of Yeungnam Medical Science
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    • v.5 no.2
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    • pp.129-139
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    • 1988
  • Vaginitis is one of the most common disease in gynecologic field in recent days. About 90% of these patients suffer from the infection of the vagina caused by Gardnerella vaginalis, Candida, or Trichomonas, either alone or in combination. For the effective diagnosis and management, it is essential to get an accurate identification of the causative agent. Applying simple and easy diagnostic methods such as pH of vaginal discharge, amine test and microscopic examination of wet mount preparation of normal saline and 10% KOH to 549 cases of randomly selected patients visiting Out-patient Department of Obstetrics and Gynecology of Yeung-nam University Hospital through May 1st to Sept. 31st, 1987, the following results were obtained. 1. In the Gardnerella vaginitis vaginal pH was more than 5.0 in 81.0% of total cases, positive amine test in 62.8%, decreased lactobacilli in 77.4% & decreased WBC counts in 70.1%, In the Trichomonas vaginitis vaginal pH more than 5.0 was noted in 93.3% of the cases, negative amine test in 90%, decreased lactobacilli in 90% & increased WBC counts in 93.3%, whereas Candida vaginitis and normal groups showed vaginal pH of 3.0~4.0 in 83.2%, negative amine test in 100%, normal distribution of lactobacilli in 89.7%, normal WBC counts in 72.4%. 2. The accuracy rates of physical diagnosis by wet mount preparation of normal saline and 10% KOH revealed 26.3%, 47.5% % 70,0% in Gardnerella vaginitis, Candida vaginitis, and Trichomonas vaginitis, respectively. The vaginal pH and amine test showed 83.2% of accuracy rate in Candida vaginitis group, 60.6% in Gardnerella vaginitis group and 83.3% III Trichomonas vaginitis group. 3. In 23 cases of Gardnerella vaginitis showing vaginal pH of 3.0 or 4.0 and positive amine test, the clue cells were observed in 10% or less in 12 cases, 30% in 5 cases, 50% in 4 cases and 50% or more in 2 cases. In summary vaginal pH and amine test could be useful in screening and differentiating the different types of infectious vaginitis. Furthermore by adding microscopic examination of normal saline and 10% KOH wet mount preparation to them, they not only could obtain higher diagnostic accuracy rate but would be more valuable in selecting the patients requiring cultures to confirm the diagnoses.

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Soluble Triggering Receptor Expressed on Myeloid cells-1: Role in the Diagnosis of Pleural Effusions (흉수의 감별 진단 시 Soluble Triggering Receptor Expressed on Myeloid Cells-1 (sTREM-1)의 유용성)

  • Kim, Jung-Hyun;Park, Eun-Young;Kim, Won-Hee;Park, Woong;Jeong, Hye-Cheol;Lee, Ji-Hyun;Kim, Eun-Kyung
    • Tuberculosis and Respiratory Diseases
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    • v.62 no.4
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    • pp.290-298
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    • 2007
  • Background: The currently available diagnostic markers for pleural effusion have a limited role. The soluble triggering receptor expressed on myeloid cells-1 (sTREM-1) is a molecule recently reported to play an important role in the myeloid cell mediated inflammatory response, and is up regulated in the body fluid by bacterial or fungal products. This study examined the expression of sTREM-1 in pleural effusion. Methods: Between April 2004 and December 2005, 48 patients with pleural effusions were enrolled in this study. The pleural fluids were taken and analyzed for the total protein, glucose, lactate dehydrogenase (LDH), adenosine deaminase (ADA), and sTREM-1. Bacterial cultures and cytology tests were also performed. Results: The clinical diagnoses were 17 parapneumonic, 14 tuberculous, and 13 malignant effusions. Four patients presented with transudates. The mean ages of the parapneumonic, tuberculous and malignant effusion groups were $57.1{\pm}19.7$, $49.5{\pm}18.6$, $66.9{\pm}15.5$, and $76.0{\pm}18.1$. respectively. The level of sTREM-1 expression was significantly higher in the parapneumonic effusions ($344.0{\pm}488.7$) than in the tuberculous effusions ($81.7{\pm}56.6$) and malignant effusions ($39.3{\pm}19.6$). With a cut-off value of 55.4pg/ml, the sensitivity and specificity for a parapneumonic effusion was 70.6% and 74.1%. Conclusion: sTREM-1 expression is significantly higher in parapneumonic effusions, suggesting its potential role as an additional diagnostic marker for pleural effusions.

Orbital and Periorbital Cellulitis in Children: A Recent 7-Year Clinical Review (소아에서 안와와 안와주위 봉와직염에 관한 최근 7년간의 임상적 고찰)

  • Choi, Jeong Ho;Hon, Sun Yeong;Park, Sung Sin;Cha, Sung Ho;Kim, Eun Jin;Lee, Jin;Chang, Jin Keun
    • Pediatric Infection and Vaccine
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    • v.15 no.1
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    • pp.36-44
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    • 2008
  • Purpose : Orbital cellulitis is rare, but it could be from the serious complication of sinusitis in children. It is often difficult to distinguish periorbital cellulitis from orbital cellulitis. The purpose of this study is to describe the clinical features of orbital and periorbital cellulitis in a pediatric population and to assess the predisposing factors and their complications. Methods : Forty-one patients aged 18 years and younger who were admitted between January 2000 and December 2006 to Hanil General Hospital and Kyunghee University Hospital with orbital or periorbital cellulitis. The retrospective analyses included clinical characteristics of orbital and periorbital cellulitis, dermographics, past history, predisposing factors, clinical presentations, treatments, and complications. Results : Among 41 patients, 34 patients had periorbital cellulitis, 7 patients had orbital cellulitis. While paranasal sinus disease was the most common predisposing cause in orbital cases, skin lesion, insect bite, dacrocystitis and conjunctivitis were the common causes in periorbital cases. In comparison with periorbital cases, orbital cases had higher level of white blood cell count, erythrocyte sedimentation rate, and C-reactive protein. Blood cultures were taken in 23 patients, but only one had Staphylococcus aureus from blood. Only one case had surgical incision and drainage and recovered without complications. Conclusion : From the results of our data, when patient shows erythematous swelling of periorbital area with opthalmoplegia, chemosis and proptosis, orbital CT scan is required to make diagnosis of orbital cellulitis. Most cases of orbital cellulitis can be treated successfully without surgical intervention. If there is no clinical improvement, repeated CT scan and/or surgical intervention should be considered.

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Clinical Significance of Enterovirus in Febrile Illness of Young Children (하절기에 발열을 주소로 입원한 3개월 이하의 영아에서 장바이러스 감염)

  • Kwak, Ji-Yeon;Cho, Mi-Hyun;Kim, Sung-Eun;Kang, Suk-Ho;Kim, Mi-Ok;Ma, Sang-Hyuk;Lee, Kyu-Man
    • Pediatric Infection and Vaccine
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    • v.8 no.1
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    • pp.94-100
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    • 2001
  • Purpose : Enterovirus is a common cause of aseptic meningitis and nonspecific febrile illness in young children. During the summer and fall months, enterovirus-infected young children are frequently admitted and evaluated to rule out bacterial sepsis and/or meningitis. The purpose of this study was to evaluate the relationship between nonpolio enterovirus infection and febrile illness in infants under 3 months of age during the summer, fall months by using a stool culture to identify the presence of enterovirus. Methods : Patients included febrile infants under 3 months of age admitted to Masan Fatima Hospital for sepsis evaluation from May 1999 to September 1999. Cultures were performed from stool and Cerebrospinal fluid samples and then were tested for enterovirus infection. Viral isolation and serotype identification were performed by cell culture and immunofluorescent testing. Enteroviruses not typed by immunofluorescent testing were confirmed by reverse transcription-polymerase chain reaction. Results : A total of 44 febrile infants were enrolled; of those, 20(45%) were positive for enterovirus. Two enterovirus culture-positive infants had concomitant urinary tract infection and one had Kawasaki disease. All infants infected with an enterovirus recovered without complications. Serotype of 20 enteroviruses were isolated from stool, 3 of echovirus type 9, 1 of echovirus type 11, 1 Coxsachievirus type B4, 15 of untyped enteroviruses. One untyped enterovirus was isolated in the CSF. Conclusion : Nonpolio enterovirus infections are associated with nonspecific febrile illnesses in infants under 3 months of age.

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Studies on the Ginseng Plants(III) -Radioactive Sodium $Acetate-U-C^{14}$ Feeding Experiments- (인삼식물(人參植物)에 관한 연구(III) -동위원소화합물(洞位元素化合物) Sodium $Acetate-U-C^{14}$을 투여한 실험-)

  • Kim, Jung-Yun;Staba, E. John
    • Korean Journal of Pharmacognosy
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    • v.5 no.2
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    • pp.111-124
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    • 1974
  • The radioactive compound sodium $acetate-U-C^{14}\;(C^{14}-acetate)$ was administered to two- and four-year-old July and September American ginseng (Araliaceae, Panax quinquefolium L.) plants and cuttings. The $C^{14}-acetate$ uptake was approximately 99%. The autoradiochromatograms suggest that the saponins isolated by preparative thin-layer chromatography contained impurities, especially those isolated from the leaf and stem extracts. The root and fruit methanol extracts yielded relatively pure saponins. The large amounts of panaquilin B and its proximity to panaquilin C on preparative thin-layer plates resulted in some admixing. The average concentration (% plant dry weight) of semi-purified saponins were high in the leaves (13.8%), as compared to fruits (9.8%), stems (7.9%) and roots (6.3%). The average percentage of $C^{14}-acetate$ incorporation into panaquilins was 4.8%. The average percentage of $C^{14}-acetate$ incorporation into panaquilins B and C was higher (1.40% and 1.13%, respectively) than that into panaquilins C, (d), G-1 and G-2 (0.75%, 0.65%, 0.13% and 0.53%, respectively). Panaquilin synthesis may be depending upon the part, collection period and age of the plant. The average percentage of $C^{14}-acetate$ incorporation into panaquilin B is high in roots (0.58%) and stems (0.48%); that into panaquilins C and (d) high in leaves (0.40% and 0.45%, respectively); and that into panaquilin E high in roots and leaves (0.55% and 0.50%, respectively). Panaquilin G-2 was synthesized in all parts of plants. The panaquilins appear to be biosynthesized more actively in July than September (exception-panaquilin G-1). Panaquilins B, C and G-1 may be biosynthesized more actively in four-year-old plants and panaquilins (d) and E more actively in two-year-old plants. The results from expectance with cuttings suggest that the panaquilins are synthesized de novo in the above-ground parts of ginseng plants, and that panaquilin G-1 may be synthesized de novo in the leaf. It is known from the tissue culture studies that panaquilins are produced by leaf, stem and root callus tissues and cailus-root cultures of American and Korean ginseng plants. Panaquilins may actively be synthesized de novo in most any cell or organ of the ginseng plants. It was verified that $C^{14}-acetate$ was incorporated into the panaxadiol portions of the panaquilins of two-year-old plants (sp. act. 0.56 mmcCi/mg) and four-year-old plants $(sp.\;act.\;0.54\;m{\mu}Ci/mg)$.

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Pathogens and Prognotic Factors for Early Onset Sepsis in Very Low Birth Weight Infants (극소 저체중 출생아에서 조기 패혈증의 원인균과 예후인자)

  • Kim, Yi-Sun;Kim, Jin-Kyu;Yoo, Hye-Soo;Ahn, So-Yoon;Seo, Hyun-Ju;Choi, Seo-Heui;Park, Soo-Kyung;Jung, Yu-Jin;Kim, Myo-Jing;Jeon, Ga-Won;Koo, Soo-Hyun;Lee, Kyung-Hoon;Chang, Yun-Sil;Park, Won-Soon
    • Neonatal Medicine
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    • v.16 no.2
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    • pp.163-171
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    • 2009
  • Purpose: This study was conducted to determine the incidence, causative pathogens, risk factors and mortality for early onset sepsis in the first three days in very low birth weight infants. Methods: The medical records of 1,124 very low birth weight infants admitted to the neonatal intensive care unit of Samsung Medical Center between November 1994 and December 2008 were retrospectively reviewed. The incidence, causative pathogens, risk factors, and mortality for early onset sepsis in the first 3 days of life in very low birth weight infants were evaluated. Results: Early onset sepsis, as confirmed by positive blood cultures, was present in 17 of 1,124 infants (1.5%). Sixty-four percent of the isolated pathogens were gram-positive bacteria and 35% of the isolated pathogens were gram-negative bacteria. The dominant pathogens of early onset sepsis included Staphylococcus aureus (23.5%), Esherichia coli (23.5%), and Enterococcus (17.6%). Vaginal delivery (adjusted odds ratio [OR], 3.7; 95% confidence interval [CI], 1.3-10.3; P=0.01) was associated with early onset sepsis. The overall mortality (adjusted hazard ratio, 3.0; 95% CI, 1.4-6.5; adjusted P=0.0039) and mortality within 72 hours of life (adjusted hazard ratio, 6.5; 95% CI, 2.2-18.9; adjusted P=0.0005) of infants with early onset sepsis were higher than that of uninfected infants. Conclusion: Early onset sepsis remains an uncommon, but potentially lethal problem among very low birth weight infants. Knowledge of the likely causative organisms and risk factors for early onset sepsis can aid in instituting prompt and appropriate therapy, in order to minimize mortality.