Author:
- INTRODUCTION
- MATERIALS AND METHODS
- Construction of DNA vaccine vector containing 56-kD gene (pKarp56).
- Confirmation of expression of 56-kDa antigen in human kidney cell line 293.
- Indirect fluorescence assay (IFA).
- Western blot analysis.
- Immunization and challenge of mice.
- Monitor antibody responses by enzyme-linked immunosorbent assay (ELISA).
- RESULTS
- DISCUSSION
ProCite
RefWorks
Reference Manager
BibTeX
Zotero
EndNote
-
1
Brown GW, Robinson DM, Huxsoll DL, Ng TS, Lim KJ, Sannasey G, 1976. Scrub typhus: a common cause of illness in indigenous populations. Trans R Soc Trop Med Hyg 70 :444–448.
-
2
Brown GW, Saunders JP, Singh S, Huxsoll DL, Shirai A, 1978. Single dose doxycycline therapy for scrub typhus. Trans R Soc Trop Med Hyg 72 :412–416.
-
3
Watt G, Chouriyagune C, Ruangweerayud R, Watcharapichat P, Phulsuksombati D, Jongsakul K, Teja-Isavadham P, Bhodhidatta K, Corcoran KD, Dasch GA, Strickman D, 1996. Scrub typhus poorly responsive to antibiotics in northern Thailand. Lancet 348 :86–89.
-
4
Watt G, Kantipong P, Jongsakul K, Watcharapichat P, Phulsuksombati D, Strickman D, 2000. Doxycycline and rifampicin for mild scrub-typhus infections in northern Thailand: a randomised trial. Lancet 356 :1057–1061.
-
5
Mathai E, Rolain JM, Verghese GM, Abraham OC, Mathai D, Mathai M, Raoult D, 2003. Outbreak of scrub typhus in southern India during the cooler months. Ann N Y Acad Sci 990 :359–364.
-
6
Smadel J, Ley HL Jr, Diercks FH, Traub R, Tipton VJ, Frick LP, 1951. Immunization against scrub typhus. I. Combined living vaccine and chemoprophylaxis in volunteers. Am J Hyg 53 :317–325.
-
7
Smadel JE, Ley HL Jr, Diercks FH, Paterson PY, Wisseman CL Jr, Traub R, 1952. Immunization against scrub typhus: duration of immunity in volunteers following combined living vaccine and chemoprophylaxis. Am J Trop Med Hyg 1 :87–99.
-
8
Eisenberg GH Jr, Osterman JV, 1979. Gamma-irradiated scrub typhus immunogens: broad-spectrum immunity with combinations of rickettsial strains. Infect Immun 26 :131–136.
-
9
Smadel JE, Ley HL Jr, Diercks FH, Traub R, 1950. Immunity in scrub typhus: resistance to induced reinfection. Arch Pathol 50 :847–861.
-
10
Eisenberg GH Jr, Osterman JV, 1978. Effects of temperature on the stability of Rickettsia tsutsugamushi and gamma-irradiated scrub typhus immunogens. Infect Immun 22 :298–300.
-
11
Seong SY, Huh MS, Jang WJ, Park SG, Kim JG, Woo SG, Choi MS, Kim IS, Chang WH, 1997. Induction of homologous immune response to Rickettsia tsutsugamushi Boryong with partial 56-kilodalton recombinant antigen fused with the maltose-binding protein MBP-Bor56. Infect Immun 65 :1541–1545.
-
12
Seong SY, Kim HR, Huh MS, Park SG, Kang JS, Han TH, Choi MS, Chang WH, Kim IS, 1997. Induction of neutralizing antibody in mice by immunization with recombinant 56 kDa protein of Orientia tsutsugamushi. Vaccine 15 :1741–1747.
-
13
Seong SY, Kim MK, Lee SM, Odgerel Z, Choi MS, Han TH, Kim IS, Kang JS, Lim BU, 2000. Neutralization epitopes on the antigenic domain II of the Orientia tsutsugamushi 56-kDa protein revealed by monoclonal antibodies. Vaccine 19 :2–9.
-
14
Ching WM, Wang H, Eamsila C, Kelly DJ, Dasch GA, 1998. Expression and refolding of truncated recombinant major outer membrane protein antigen (r56) of Orientia tsutsugamushi and its use in an enzyme-linked immunosorbent assay. Clin Diagn Lab Immunol 5 :519–526.
-
15
Ching WM, Rowland D, Zhang Z, Bourgeois AL, Kelly D, Dasch GA, Devine PL, 2001. Early diagnosis of scrub typhus with a rapid flow assay using recombinant major outer membrane protein antigen (r56) of Orientia tsutsugamushi. Clin Diagn Lab Immunol 8 :409–414.
-
16
Kim IS, Seong SY, Woo SG, Choi MS, Kang JS, Chang WH, 1993. Rapid diagnosis of scrub typhus by a passive hemagglutination assay using recombinant 56-kDa polypeptides. J Clin Microbiol 31 :2057–2060.
-
17
Jang WJ, Huh MS, Park KH, Choi MS, Kim IS, 2003. Evaluation of an immunoglobulin M capture enzyme-linked immunosorbent assay for diagnosis of Orientia tsutsugamushi infection. Clin Diagn Lab Immunol 10 :394–398.
-
18
Hanson BA, 1983. Effect of immune serum on infectivity of Rickettsia tsutsugamushi. Infect Immun 42 :341–349.
-
19
Chattopadhyay S, Jiang J, Chan T-C, Chao CC, Ching W-M, Richards AL, 2005. Successful induction of humoral and cellular immune responses in cynomolgus monkeys to the scrub typhus vaccine candidate, Kp r56. Infect Immun 73 :5039–5047.
-
20
Nagata T, Aoshi T, Uchijima M, Suzuki M, Koide Y, 2004. Cytotoxic T-lymphocyte and helper T-lymphocyte-oriented DNA vaccination. DNA Cell Biol 23 :93–106.
-
21
Putnak R, Porter K, Schmaljohn C, 2003. DNA vaccines for flaviviruses. Adv Virus Res 61 :445–468.
-
22
Ada G, Ramshaw I, 2003. DNA vaccination. Expert Opin Emerg Drugs 8 :27–35.
-
23
Wolff JA, Malone RW, Williams P, Chong W, Acsadi G, Jani A, Felgner PL, 1990. Direct gene transfer into mouse muscle in vivo. Science 247 :1465–1468.
-
24
Ulmer JB, Donnelly JJ, Parker SE, Rhodes GH, Felgner PL, Dwarki VJ, Gromkowski SH, Deck RR, DeWitt CM, Friedman A, 1993. Heterologous protection against influenza by injection of DNA encoding a viral protein. Science 259 :1745.
-
25
Sedegah MR, Hedstrom R, Hobart P, Hoffman SL, 1994. Protection against malaria by immunization with plasmid DNA encoding circumsporozoite protein. Proc Natl Acad Sci USA 91 :9866–9870.
-
26
Cohen J, 1993. Naked DNA points way to vaccine. Science 259 :1691–1692.
-
27
Ohashi N, Nashimoto H, Ikeda H, Tamura A, 1992. Diversity of immunodominant 56-kDa type-specific antigen (TSA) of Rickettsia tsutsugamushi: sequence and comparative analyses of the genes encoding TSA homologues from four antigenic variants. J Biol Chem 267 :12728–12735.
-
28
Enatsu T, Urakami H, Tamura A, 1999. Phylogenetic analysis of Orientia tsutsugamushi strains based on the sequence homologies of 56-kDa type-specific antigen genes. FEMS Microbiol Lett 180 :163–169.
-
29
Stover CK, Marana DP, Carter JM, Roe BA, Mardis E, Oaks EV, 1990. The 56-kilodalton major protein antigen of Rickettsia tsutsugamushi: molecular cloning and sequence analysis of the sta56 gene and precise identification of a strain-specific epitope. Infect Immun 58 :2076–2084.
-
30
Sambrook J, Fritsch E, Maniatis T, 1989. Molecular Cloning: A Laboratory Manual. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press.
-
31
Moree MF, Hanson B, 1992. Growth characteristics and proteins of plaque-purified strains of Rickettsia tsutsugamushi. Infect Immun 60 :3405–3415.
-
32
Laemmli UK, 1970. Cleavage of structural proteins during assembly of the head of bacteriophage T4. Nature 227 :680.
-
33
Chan TC, Jiang J, Temenak JJ, Richards AL, 2003. Development of a rapid method for determining the infectious dose (ID)50 of Orientia tsutsugamushi in a scrub typhus mouse model for the evaluation of vaccine candidates. Vaccine 21 :4550–4554.
-
34
Jerrells TR, Osterman JV, 1982. Host defenses in experimental scrub typhus: deplayed-type hypersensitivity responses of inbred mice. Infect Immun 35 :117–123.
-
35
Jerrells TR, Palmer BA, Osterman JV, 1983. Gamma-irradiated scrub typhus immunogens: development of cell-mediated immunity after vaccination of inbred mice. Infect Immun 39 :262–269.
-
36
de Arruda LB, Chikhlikar PR, August JT, Marques ET, 2004. DNA vaccine encoding human immunodeficiency virus-1 Gag, targeted to the major histocompatibility complex II compartment by lysosomal-associated membrane protein, elicits enhanced long-term memory response. Immunology 112 :126–133.
-
37
Mwangi W, Brown WC, Lewin HA, Howard CJ, Hope JC, Baszler TV, Caplazi P, Abbott J, Palmer GH, 2002. DNA-encoded fetal liver tyrosine kinase 3 ligand and granulocyte macrophage-colony-stimulating factor increase dendritic cell recruitment to the inoculation site and enhance antigen-specific CD4+ T cell responses induced by DNA vaccination of out-bred animals. J Immunol 69 :3837–3846.
-
38
Raviprakash K, Kochel TJ, Ewing D, Simmons M, Phillips I, Hayes CG, Porter KR, 2000. Immunogenicity of dengue virus type 1 DNA vaccines expressing truncated and full length envelope protein. Vaccine 18 :2426–2434.
-
39
McNeel DG, Schiffman K, Disis ML, 1999. Immunization with recombinant human granulocyte-macrophage colony-stimulating factor as a vaccine adjuvant elicits both a cellular and humoral response to recombinant human granulocyte-macrophage colony-stimulating factor. Blood 93 :2653–2659.
-
40
Stobie L, Gurunathan S, Prussin C, Sacks DL, Glaichenhaus N, Wu CY, Seder RA, 2000. The role of antigen and IL-12 in sustaining Th1 memory cells in vivo: IL-12 is required to maintain memory/effector Th1 cells sufficient to mediate protection to an infectious parasite challenge. Proc Natl Acad Sci USA 97 :8427–8432.
-
41
Rao M, Matyas GR, Grieder F, Anderson K, Jahrling PB, Alving CR, 1999. Cytotoxic T lymphocytes to Ebola Zaire virus are induced in mice by immunization with liposomes containing lipid A. Vaccine 17 :2991–2998.
| Past two years | Past Year | Past 30 Days | |
|---|---|---|---|
| Abstract Views | 19 | 18 | 5 |
| Full Text Views | 305 | 86 | 0 |
| PDF Downloads | 66 | 16 | 0 |
PROTECTION AGAINST SCRUB TYPHUS BY A PLASMID VACCINE ENCODING THE 56-KD OUTER MEMBRANE PROTEIN ANTIGEN GENE
YI-SHENG NI
YI-SHENG NI
Rickettsial Diseases Department, U.S. Naval Medical Research Center, Silver Spring, Maryland; Uniformed Services University of the Health Sciences, Bethesda, Maryland
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TEIK-CHYE CHAN
TEIK-CHYE CHAN
Rickettsial Diseases Department, U.S. Naval Medical Research Center, Silver Spring, Maryland; Uniformed Services University of the Health Sciences, Bethesda, Maryland
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CHIEN-CHUNG CHAO
CHIEN-CHUNG CHAO
Rickettsial Diseases Department, U.S. Naval Medical Research Center, Silver Spring, Maryland; Uniformed Services University of the Health Sciences, Bethesda, Maryland
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ALLEN L. RICHARDS
ALLEN L. RICHARDS
Rickettsial Diseases Department, U.S. Naval Medical Research Center, Silver Spring, Maryland; Uniformed Services University of the Health Sciences, Bethesda, Maryland
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GREGORY A. DASCH
GREGORY A. DASCH
Rickettsial Diseases Department, U.S. Naval Medical Research Center, Silver Spring, Maryland; Uniformed Services University of the Health Sciences, Bethesda, Maryland
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WEI-MEI CHING
WEI-MEI CHING
Rickettsial Diseases Department, U.S. Naval Medical Research Center, Silver Spring, Maryland; Uniformed Services University of the Health Sciences, Bethesda, Maryland
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The 56-kD outer membrane protein of Orientia tsutsugamushi has previously been shown to be the immunodominant antigen in scrub typhus infections. Its gene was cloned into the DNA vaccine vector pVR1012 as a vaccine candidate (pKarp56). The in vitro expression of this 56-kD antigen by pKarp56 was confirmed in tissue culture by an indirect fluorescence assay and Western blot analysis. The initial antibody responses of mice immunized with varied doses of the pKarp56 were barely detected, but increases were observed after each of three subsequent booster immunizations. Although no protection was observed with a single immunization of pKarp56, after four immunizations, 60% of the mice survived a 1,000 × 50% lethal dose (LD50) challenge. These results specifically confirm the importance of the 56-kD protein antigen in protective immunity against O. tsutsugamushi and demonstrate the feasibility of DNA vaccines for the prevention of scrub typhus.
- INTRODUCTION
- MATERIALS AND METHODS
- Construction of DNA vaccine vector containing 56-kD gene (pKarp56).
- Confirmation of expression of 56-kDa antigen in human kidney cell line 293.
- Indirect fluorescence assay (IFA).
- Western blot analysis.
- Immunization and challenge of mice.
- Monitor antibody responses by enzyme-linked immunosorbent assay (ELISA).
- RESULTS
- DISCUSSION
ProCite
RefWorks
Reference Manager
BibTeX
Zotero
EndNote
-
1
Brown GW, Robinson DM, Huxsoll DL, Ng TS, Lim KJ, Sannasey G, 1976. Scrub typhus: a common cause of illness in indigenous populations. Trans R Soc Trop Med Hyg 70 :444–448.
-
2
Brown GW, Saunders JP, Singh S, Huxsoll DL, Shirai A, 1978. Single dose doxycycline therapy for scrub typhus. Trans R Soc Trop Med Hyg 72 :412–416.
-
3
Watt G, Chouriyagune C, Ruangweerayud R, Watcharapichat P, Phulsuksombati D, Jongsakul K, Teja-Isavadham P, Bhodhidatta K, Corcoran KD, Dasch GA, Strickman D, 1996. Scrub typhus poorly responsive to antibiotics in northern Thailand. Lancet 348 :86–89.
-
4
Watt G, Kantipong P, Jongsakul K, Watcharapichat P, Phulsuksombati D, Strickman D, 2000. Doxycycline and rifampicin for mild scrub-typhus infections in northern Thailand: a randomised trial. Lancet 356 :1057–1061.
-
5
Mathai E, Rolain JM, Verghese GM, Abraham OC, Mathai D, Mathai M, Raoult D, 2003. Outbreak of scrub typhus in southern India during the cooler months. Ann N Y Acad Sci 990 :359–364.
-
6
Smadel J, Ley HL Jr, Diercks FH, Traub R, Tipton VJ, Frick LP, 1951. Immunization against scrub typhus. I. Combined living vaccine and chemoprophylaxis in volunteers. Am J Hyg 53 :317–325.
-
7
Smadel JE, Ley HL Jr, Diercks FH, Paterson PY, Wisseman CL Jr, Traub R, 1952. Immunization against scrub typhus: duration of immunity in volunteers following combined living vaccine and chemoprophylaxis. Am J Trop Med Hyg 1 :87–99.
-
8
Eisenberg GH Jr, Osterman JV, 1979. Gamma-irradiated scrub typhus immunogens: broad-spectrum immunity with combinations of rickettsial strains. Infect Immun 26 :131–136.
-
9
Smadel JE, Ley HL Jr, Diercks FH, Traub R, 1950. Immunity in scrub typhus: resistance to induced reinfection. Arch Pathol 50 :847–861.
-
10
Eisenberg GH Jr, Osterman JV, 1978. Effects of temperature on the stability of Rickettsia tsutsugamushi and gamma-irradiated scrub typhus immunogens. Infect Immun 22 :298–300.
-
11
Seong SY, Huh MS, Jang WJ, Park SG, Kim JG, Woo SG, Choi MS, Kim IS, Chang WH, 1997. Induction of homologous immune response to Rickettsia tsutsugamushi Boryong with partial 56-kilodalton recombinant antigen fused with the maltose-binding protein MBP-Bor56. Infect Immun 65 :1541–1545.
-
12
Seong SY, Kim HR, Huh MS, Park SG, Kang JS, Han TH, Choi MS, Chang WH, Kim IS, 1997. Induction of neutralizing antibody in mice by immunization with recombinant 56 kDa protein of Orientia tsutsugamushi. Vaccine 15 :1741–1747.
-
13
Seong SY, Kim MK, Lee SM, Odgerel Z, Choi MS, Han TH, Kim IS, Kang JS, Lim BU, 2000. Neutralization epitopes on the antigenic domain II of the Orientia tsutsugamushi 56-kDa protein revealed by monoclonal antibodies. Vaccine 19 :2–9.
-
14
Ching WM, Wang H, Eamsila C, Kelly DJ, Dasch GA, 1998. Expression and refolding of truncated recombinant major outer membrane protein antigen (r56) of Orientia tsutsugamushi and its use in an enzyme-linked immunosorbent assay. Clin Diagn Lab Immunol 5 :519–526.
-
15
Ching WM, Rowland D, Zhang Z, Bourgeois AL, Kelly D, Dasch GA, Devine PL, 2001. Early diagnosis of scrub typhus with a rapid flow assay using recombinant major outer membrane protein antigen (r56) of Orientia tsutsugamushi. Clin Diagn Lab Immunol 8 :409–414.
-
16
Kim IS, Seong SY, Woo SG, Choi MS, Kang JS, Chang WH, 1993. Rapid diagnosis of scrub typhus by a passive hemagglutination assay using recombinant 56-kDa polypeptides. J Clin Microbiol 31 :2057–2060.
-
17
Jang WJ, Huh MS, Park KH, Choi MS, Kim IS, 2003. Evaluation of an immunoglobulin M capture enzyme-linked immunosorbent assay for diagnosis of Orientia tsutsugamushi infection. Clin Diagn Lab Immunol 10 :394–398.
-
18
Hanson BA, 1983. Effect of immune serum on infectivity of Rickettsia tsutsugamushi. Infect Immun 42 :341–349.
-
19
Chattopadhyay S, Jiang J, Chan T-C, Chao CC, Ching W-M, Richards AL, 2005. Successful induction of humoral and cellular immune responses in cynomolgus monkeys to the scrub typhus vaccine candidate, Kp r56. Infect Immun 73 :5039–5047.
-
20
Nagata T, Aoshi T, Uchijima M, Suzuki M, Koide Y, 2004. Cytotoxic T-lymphocyte and helper T-lymphocyte-oriented DNA vaccination. DNA Cell Biol 23 :93–106.
-
21
Putnak R, Porter K, Schmaljohn C, 2003. DNA vaccines for flaviviruses. Adv Virus Res 61 :445–468.
-
22
Ada G, Ramshaw I, 2003. DNA vaccination. Expert Opin Emerg Drugs 8 :27–35.
-
23
Wolff JA, Malone RW, Williams P, Chong W, Acsadi G, Jani A, Felgner PL, 1990. Direct gene transfer into mouse muscle in vivo. Science 247 :1465–1468.
-
24
Ulmer JB, Donnelly JJ, Parker SE, Rhodes GH, Felgner PL, Dwarki VJ, Gromkowski SH, Deck RR, DeWitt CM, Friedman A, 1993. Heterologous protection against influenza by injection of DNA encoding a viral protein. Science 259 :1745.
-
25
Sedegah MR, Hedstrom R, Hobart P, Hoffman SL, 1994. Protection against malaria by immunization with plasmid DNA encoding circumsporozoite protein. Proc Natl Acad Sci USA 91 :9866–9870.
-
26
Cohen J, 1993. Naked DNA points way to vaccine. Science 259 :1691–1692.
-
27
Ohashi N, Nashimoto H, Ikeda H, Tamura A, 1992. Diversity of immunodominant 56-kDa type-specific antigen (TSA) of Rickettsia tsutsugamushi: sequence and comparative analyses of the genes encoding TSA homologues from four antigenic variants. J Biol Chem 267 :12728–12735.
-
28
Enatsu T, Urakami H, Tamura A, 1999. Phylogenetic analysis of Orientia tsutsugamushi strains based on the sequence homologies of 56-kDa type-specific antigen genes. FEMS Microbiol Lett 180 :163–169.
-
29
Stover CK, Marana DP, Carter JM, Roe BA, Mardis E, Oaks EV, 1990. The 56-kilodalton major protein antigen of Rickettsia tsutsugamushi: molecular cloning and sequence analysis of the sta56 gene and precise identification of a strain-specific epitope. Infect Immun 58 :2076–2084.
-
30
Sambrook J, Fritsch E, Maniatis T, 1989. Molecular Cloning: A Laboratory Manual. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press.
-
31
Moree MF, Hanson B, 1992. Growth characteristics and proteins of plaque-purified strains of Rickettsia tsutsugamushi. Infect Immun 60 :3405–3415.
-
32
Laemmli UK, 1970. Cleavage of structural proteins during assembly of the head of bacteriophage T4. Nature 227 :680.
-
33
Chan TC, Jiang J, Temenak JJ, Richards AL, 2003. Development of a rapid method for determining the infectious dose (ID)50 of Orientia tsutsugamushi in a scrub typhus mouse model for the evaluation of vaccine candidates. Vaccine 21 :4550–4554.
-
34
Jerrells TR, Osterman JV, 1982. Host defenses in experimental scrub typhus: deplayed-type hypersensitivity responses of inbred mice. Infect Immun 35 :117–123.
-
35
Jerrells TR, Palmer BA, Osterman JV, 1983. Gamma-irradiated scrub typhus immunogens: development of cell-mediated immunity after vaccination of inbred mice. Infect Immun 39 :262–269.
-
36
de Arruda LB, Chikhlikar PR, August JT, Marques ET, 2004. DNA vaccine encoding human immunodeficiency virus-1 Gag, targeted to the major histocompatibility complex II compartment by lysosomal-associated membrane protein, elicits enhanced long-term memory response. Immunology 112 :126–133.
-
37
Mwangi W, Brown WC, Lewin HA, Howard CJ, Hope JC, Baszler TV, Caplazi P, Abbott J, Palmer GH, 2002. DNA-encoded fetal liver tyrosine kinase 3 ligand and granulocyte macrophage-colony-stimulating factor increase dendritic cell recruitment to the inoculation site and enhance antigen-specific CD4+ T cell responses induced by DNA vaccination of out-bred animals. J Immunol 69 :3837–3846.
-
38
Raviprakash K, Kochel TJ, Ewing D, Simmons M, Phillips I, Hayes CG, Porter KR, 2000. Immunogenicity of dengue virus type 1 DNA vaccines expressing truncated and full length envelope protein. Vaccine 18 :2426–2434.
-
39
McNeel DG, Schiffman K, Disis ML, 1999. Immunization with recombinant human granulocyte-macrophage colony-stimulating factor as a vaccine adjuvant elicits both a cellular and humoral response to recombinant human granulocyte-macrophage colony-stimulating factor. Blood 93 :2653–2659.
-
40
Stobie L, Gurunathan S, Prussin C, Sacks DL, Glaichenhaus N, Wu CY, Seder RA, 2000. The role of antigen and IL-12 in sustaining Th1 memory cells in vivo: IL-12 is required to maintain memory/effector Th1 cells sufficient to mediate protection to an infectious parasite challenge. Proc Natl Acad Sci USA 97 :8427–8432.
-
41
Rao M, Matyas GR, Grieder F, Anderson K, Jahrling PB, Alving CR, 1999. Cytotoxic T lymphocytes to Ebola Zaire virus are induced in mice by immunization with liposomes containing lipid A. Vaccine 17 :2991–2998.
| Past two years | Past Year | Past 30 Days | |
|---|---|---|---|
| Abstract Views | 19 | 18 | 5 |
| Full Text Views | 305 | 86 | 0 |
| PDF Downloads | 66 | 16 | 0 |