臺灣博碩士論文加值系統

陰道鞭毛蟲為WHO宣佈為最廣泛的非病毒性性接觸傳染疾病，已證實會造成HIV感染機率增加、與子宮頸癌、不孕症有關聯性，且其細胞週期的G2期較G1期長，但對陰道鞭毛蟲的仍不清楚。之前曾將陰道鞭毛蟲培養於低糖的卡路里限制液態培養基，發現會增加其存活時間，本研究想研究陰道鞭毛蟲於卡路里限制時其轉錄體基因調控機制，利用之前所建立的三個基因表達序列資料庫︰TvEST（5934條序列），TvCS（9941條序列）及TvG2M（10687條序列），進行複製擴增得到7668非重複性序列當作架構陰道鞭毛蟲微陣列晶片之探針。以卡路里限制培養，選擇延滯期、對數期及生長穩定期不同生長時期進行微陣列晶片分析，發現與卡路里限制相關的基因，明顯減少的有Hydrogenosomal malic enzyme (AP65-1)、Cyclophilin type 5、hypothetical proteins及MAPKKK，明顯增加的有28S ribosomal RNA protein, 40S ribosomal protein S3Ae, zzz-NO HIT與不同的hypothetical proteins，再以RT-PCR加以證實微陣列晶片之實驗結果。推論在卡路里限制時陰道鞭毛蟲之DNA複製、protein turnover相關基因是表達量增加的；蛋白質合成、醣解作用、產生能量相關基因是表達量降低的。

Trichomonas vaginalis is one of the most common non-viral sexually transmitted disease, and trichomoniasis is emerging as one of the most important cofactors in amplifying HIV transmission, also linked to this disease are cervical cancer and infertility. However, very little is known about the biology of T. vaginalis at molecular level. We aimed to study the effect of calorie-restriction on the growth of this unicellular parasite. Previous studies showed that calorie restriction can increase the life-span of many model organisms. A T. vaginalis cDNA microarray (TvARRAY V1) was fabricated by using 7668 nonredundant expressed sequence tags (EST) selected from three cDNA libraries: TvEST(5934 sequences), TvCS(9941 sequences) and TvG2M(10687 sequences).At lag、logarith- mic and stationary phases under calorie-restriction comparative micro- array analysis reveals calorie-restriction related genes including, downre- gulation：Hydrogenosomal malic enzyme (AP65-1),cyclophilin type 5, MAPKKK and Hypothetical proteins；upregulation：28S ribosomal RNA protein, 40S Ribosomal protein S3Ae, zzz-NO HIT and hypothetical pro- teins. Reverse transcription PCR analysis of selected genes also showed similar results, thereby supporting the reliability of the TvARRAY system. Thereby,calorie-restriction produced changes in gene expression consistent with increased DNA replication and protein turnover related genes；which decreased protein synthesis、glycolysis and energy production related genes.

摘要---------------------------------------------------------------------------------1
§ 第一章 序論--------------------------------------------------------------------4
1.1前言-----------------------------------------------------------------------------4
1.2 陰道鞭毛蟲外部型態超及微架構----------------------------------------5
1.3生理特性-----------------------------------------------------------------------6
1.4 陰道鞭毛蟲生活史與傳染途徑-------------------------------------------7
1.5 表現序列標籤Expressed Sequence Tag----------------------------------7
1.6 互補核酸微陣列晶片（cDNA microarray）------------------------------8
1.7 微陣列晶片實驗條件之選擇----------------------------------------------9
1.7.1 與CR相關基因----------------------------------------------------------10
1.7.2 CR in Saccharomyces cerevisiae ---------------------------------------10
1.7.3 CR in Drosophila---------------------------------------------------------11
1.7.4 CR in Mice-----------------------------------------------------------------11
1.7.5 CR in Caenorhabditis elegans------------------------------------------12
1.7.6 CR in Leishmania major ------------------------------------------------13
1.7.7 Glycolytic enzymes------------------------------------------------------14
1.7.7.1 Hexokinase--------------------------------------------------------------14
1.7.7.2 Lactate dehydrogenase-------------------------------------------------15
1.7.7.3 Glyceraldehyde-3-phosphate dehydrogenase ----------------------16
1.7.7.4 Enolase-------------------------------------------------------------------16
1.7.7.5 Glucose-6-phosphate isomerase--------------------------------------17
1.7.8 以卡路里限制做為實驗------------------------------------------------17

§第二章 材料方法--------------------------------------------------------------18
2.1 細胞株與培養基-----------------------------------------------------------18
2.2 陰道鞭毛蟲的觀察與計數-----------------------------------------------18
2.3緩沖液和溶液---------------------------------------------------------------18
2.4 核酸引子----------------------------------------------------------19
2.5連鎖聚合酶反應(Polymerase Chain Reaction)-------------------------19
2.6連鎖聚合酶反應產物純化------------------------------------------------19
2.7 核酸洋菜膠電泳分析（Agarose gel electrophoresis）-----------------20
2.8 以點接觸法藉點片裝置（Q Array, Genetix）將探針點到晶片上--------------------------------------------------------------------------------------20
2.9 陰道鞭毛蟲內總RNA的萃取----------------------------------------21
2.10 陰道鞭毛蟲內總RNA的純化------------------------------------------22
2.11 Agilent 2100 bioanalyzer測量RNA濃度------------------------------22
2.12. 微陣列雜交反應-------------------------------------------------------23
2.13微陣列結果分析-------------------------------------------------------26
2.14 反轉錄連鎖聚合酶反應-------------------------------------------------27

§第三章 結果--------------------------------------------------------------------28
3.1 微陣列晶片探針之挑選-------------------------------------------------29
3.2 微陣列晶片探針--------------------------------------------------------29
3.3 陰道鞭毛蟲之低糖培養--------------------------------------------------29
3.4 比較不同反轉錄酶primer對微陣列晶片雜交反應的影響---------30
3.5 比較不同反轉錄primer對微陣列晶片雜交的影響之實驗結果分 析----------------------------------------------------------------------------31
3.6 陰道鞭毛蟲不同低糖培養時間點之微陣列晶片雜交反應--------32
3.6.1 DNA複製------------------------------------------------------------------33
3.6.2 轉錄作用------------------------------------------------------------------34
3.6.3.1 轉譯作用----------------------------------------------------------------34
3.6.3.2 Protein turnover---------------------------------------------------------35
3.6.4 醣解作用------------------------------------------------------------------36
3.6.5 訊號傳遞------------------------------------------------------------------36
3.6.6 Lipid----------------------------------------------------------------------37
3.6.7 能量相關基因或蛋白質------------------------------------------------37
3.6.8 其他------------------------------------------------------------------------38
3.7 RT-PCR-----------------------------------------------------------------------39
§第四章 討論與結論-----------------------------------------------------------40
§參考文獻------------------------------------------------------------------------48
§附圖與表格---------------------------------------------------------------------61

1.Abonyi, A. (1995) Examination of nonflagellate and flagellate round forms of Trichomonas vaginalis by transmission electron microscopy. Appl. Parasitol. 36:303–310.
2.Ahuatzi, D., Herrero, P., de la Cera, T., and Moreno, F. (2004) The glucose-regulated nuclear localization of hexokinase 2 in Saccharomyces cerevisiae is Mig1-dependent. J. Biol. Chem. 279: 14440–14446.
3.Anderson, R.M., Bitterman, K.J., Wood, J.G., Medvedik, O., Cohen, H., Lin, S.S., Manchester, J.K., Gordon, J.I., and Sinclair, D.A. (2002) Manipulation of a nuclear NAD+ salvage pathway delays aging without altering steady-state NAD+ levels. J. Biol. Chem. 277: 18881–18890.
4.Anderson, R.M., Bitterman, K.J., Wood, J.G., Medvedik, O., and Sinclair, D.A. (2003) Nicotinamide and PNC1 govern lifespan extension by calorie restriction in Saccharomyces cerevisiae. Nature 423:181–185.
5.Arese, P., and Cappuccinelli, P. (1974) Glycolysis and the pentose phosphate cycle in Trichomonas vaginalis. I. Enzyme activity pattern and the constant proportion quintet. Int. J. Biochem. 5:859–865.
6.Arroyo, R., A. Gonzalez-Robles, A. Martinez-Palomo, and John, F. A. (1993) Signalling of Trichomonas vaginalis for amoeboid transformation and adhesion synthesis follows cytoadherence. Mol. Microbiol. 7:299–309.
7.Baerenstein, H. D. (1961) Lactic dehydrogenase in Trichomonas vaginalis. J. Parasitol. 47:279–284.
8.Baptiste, V., Laurent, V., Ali, O., and Denis, S. (2005) Stage-specific antileishmanial activity of an inhibitor of SIR2 histone deacetylase. Acta. Tropica. 94:107-115.
9.Benchimol, M., and de Souza, W. (1995) Carbohydrate involvement in the association of a prokaryotic cell with Trichomonas vaginalis and Tritrichomonas foetus. Parasitol. Res. 81:459–464.
10.Benchimol, M., Johnson, P. J., and de Souza, W. (1996) Morphoge- nesis of the hydrogenosome: an ultrastructural study. Biol. Cell 87:197–205.
11.Berrigan, D., Perkins, S., Haines, D.C., and Hursting, S. (2002) Adult-onset calorie restriction and fasting delay the spontaneous tumorigenesis in p53-deficient mice. Carcinogenesis 23:817–822.
12.Bilel Y., Ali T., and Ali O.(2005) A Leishmania major protein with
extensive homology to silent information regulator 2 of Saccharo- myces cerevisiae.Gene 169:115-118.
13.Blüher, M., Kahn, B.B., and Kahn, C.R. (2003) Extended longevity in mice lacking the insulin receptor in adipose tissue. Science 299: 572–574.
14.Brown, M. T. (1972) Trichomoniasis. Practitioner 209:639-644.
15.Brunet, A., Sweeney, L.B., Sturgill, J.F., Chua, K.F., Greer, P.L., Lin, Y., Tran, H., Ross, S.E., Mostoslavsky, R., and Cohen, H.Y., (2004) Stress-dependent regulation of FOXO transcription factors by the SIRT1 deacetylase. Science 303:2011–2015.
16.Bryson, J.M. et al. (2002) Increased hexokinase activity, of either
ectopic or endogenous origin, protects renal epithelial cells against
acute oxidant-induced cell death. J. Biol. Chem. 277:11392–11400.
17.Cameron, D. W., and Padian, N. S. (1990) Sexual transmission of HIV and the epidemiology of other sexually transmitted diseases. AIDS 4(Suppl. 1):S99–S103.
18.Cappuccinelli, P., Sellitto, C., Zicconi, D., and Juliano, C. (1987) Structural and molecular organization of Trichomonas vaginalis cytoskeleton. Acta Univ. Carol. Biol. 30:211–217.
19.Catterall, R. D., and Nicol, C. S. (1960) Is trichomonal infestation a venereal disease? Br. Med. J. 1: 1177.
20.Chapman, A., Linstead, D., Lloyd, D., and Williams, J. (1987) 13C-NMR reveals glycerol as a unexpected major metabolite of the protozoan parasite Trichomonas vaginalis. FEBS Lett. 191:287–292.
21.Chiu, C. H., Lin, W. D., Huang, S. Y., and Lee, Y.H. (2004) Effect of a C/EBP gene replacement on mitochondrial biogenesis in fat cells. Genes Dev. 18:1970–1975.
22.Cheol-Koo L., Richard, W., Tomoas, A. P. (2000) Gene-expression profile of the aging brain in mice. Nature genetics 25:294-297.
23.Chose, O., Noel, C., Gerbod, D., Brenner, C., Viscogliosi, E., and Roseto, A. (2002) A form of cell death with some features resembling apoptosis in the amitochondrial unicellular organism Trichomonas vaginalis. Exp. Cell Res. 276:32-9.
24.Clancy, D.J., Gems, D., Harshman, L.G., Oldham, S., Stocker, H.,
Hafen, E., Leevers, S.J., and Partridge, L. (2001) Extension of lifespan to CR in these mice as a first approximation of span by loss of CHICO, a Drosophila insulin receptor substrate protein. Science 292:104–106.
25.Cohen, H.Y., Miller, C., Bitterman, K.J., Wall, N.R., Hekking, B., Kessler, B., Howitz, K.T., Gorospe, M., de Cabo, R., and Sinclair, D.A. (2004) Calorie restriction promotes mammalian cell survival by inducing the SIRT1 deacetylase. Science 305:390–392.
26.Cooper, J.A. et al. (1984) Phosphorylation sites in enolase and lactate
dehydrogenase utilized by tyrosine protein kinases in vivo and in vitro. J. Biol. Chem. 259:7835–7841.
27.Cotch, M. F. (1990) Carriage of Trichomonas vaginalis (Tv) is associated with adverse pregnancy outcome, American Society for Microbiology, Washington, D.C. abstr. 681:199.
28.Diamond, L. S. (1986) In vitro cultivation of the Trichomonadidae: a state of the art review. Acta Univ. Carol. Biol. 30:221–228.
29.Diamond, L. S., Clark, C. G., and Cunnick, C. C. (1995) YI-S, a casein-free medium for axenic cultivation of Entamoeba histolytica, related Entamoeba, Giardia intestinalis and Trichomonas vaginalis. J. Eukaryot. Microbiol. 42: 277–278.
30.Diderich, J.A. et al. (2001) Physiological properties of Saccharomy- ces cerevisiae from which hexokinase II has been deleted. Appl. Environ. Microbiol. 67:1587–1593
31.Dino, P., Kiera, D., Renuka B., and Gray G. (1998) Clinical and Microbiological Aspects of Trichomonas vaginalis. Clin. Microb. Review :300-317.
32.de la Cera, T. et al. (2002) Mediator factor Med8p interacts with the
hexokinase 2: implication in the glucose signaling pathway of
Saccharomyces cerevisiae. J. Mol. Biol. 319:703–714.
33.Fari, A., Trevoux, R., and Verges. V. (1985) Diagnosis and significance of non-motile forms of Trichomonas vaginalis, abstr. 28a. In Abstracts of the International Symposium on trichomonads and Trichomoniasis.
34.Feo, S. et al. (2000) ENO1 gene product binds to the c-myc promoter
and acts as a transcriptional repressor: relationship with Myc promoter-binding protein 1 (MBP-1). FEBS Lett. 473:47–52.
35.Fouts, A. C., and Kraus, S. J. (1980) Trichomonas vaginalis: reevaluation of its clinical presentation and laboratory diagnosis. J. Infect. Dis. 141:137–143.
36.Frye, R. A. (2000) Phylogenetic classification of prokaryotic and eukaryotic Sir2-like proteins. Biochem.Biophys. Res. Comm. 273: 793–798.
37.Ghosh, A. K. et al. (1999) MBP-1physically associates with histone deacetylase for transcriptional repression. Biochem. Biophys. Res. Commun. 260:405-09.
38.Gottlob, K. et al. (2001) Inhibition of early apoptotic events by
Akt/PKB is dependent on the first committed step of glycolysis and
mitochondrial hexokinase. Genes Dev. 15:1406–1418.
39.Gram, I., Macaluso, M., Churchill, J. and Stalsberg, H. (1992) Trichomonas vaginalis (TV) and human papillomavirus (HPV) infection and the incidence of cervical intraepithelial neoplasia (CIN) grade III. Cancer Causes Control 3:231–236.
40.Grodstein, F., Goldman, M. B., and Cramer, D. W. (1993) Relation of tubal infertility to a history of sexually transmitted diseases. Am. J. Epidemiol. 137:577–584.
41.Grys, E. (1973) Diagnosis of trichomonadosis in the urinary tract of women (preliminary report). Wiad. Parazytol. 19:461–462. (In Polish.)
42.Hardy, P. H., Hardy, J., Nell, B. E., Graham, D. A., Spence, M. R., and Rosenbaum, R. C. (1984) Prevalence of six sexually transmitted disease agents among pregnant inner-city adolescents and pregnancy outcome. Lancet ii: 333–337.
43.Heath, J. P. (1981) Behaviour and pathogenicity of Trichomonas vaginalis in epithelial cell cultures: a study by light and scanning electron microscopy. Br. J. Vener. Dis. 57:106–117.
44.Heine, P., and McGregor, J. A. (1993) Trichomonas vaginalis: a reemerging pathogen. Clin. Obstet. Gynecol. 36:137–144.
45.Herrero, P. et al. (1995) Transcriptional regulation of the Saccharomyces cerevisiae HXK1, HXK2 and GLK1 genes. Yeast 11: 137–144
46.Herrero, P. et al. (1996) Identification and characterisation of two
transcriptional repressor elements within the coding sequence of the
Saccharomyces cerevisiae HXK2 gene. Nucleic Acids Res. :1822– 1828.
47.Herrero, P. et al. (1998) The hexokinase 2 protein participates in
regulatory DNA-protein complexes necessary for glucose repression of the SUC2 gene in Saccharomyces cerevisiae. FEBS Lett. 434:71– 76.
48.Honigberg, B. M., and King, V. M. (1964) Structure of Trichomonas vaginalis Donne. J. Parasitol. 50:345–364.
49.Honigberg, B. M., Gupta, P. K., Spence, M. R., Frost, J. K., Kuczyn- ska, Choromanski, K. L., and Warton, A. (1984) Pathogenicity of Trichomonas vaginalis:cytopathologic and histopathologic changes of the cervical epithelium. Obstet. Gynecol. 64:179–184.
50.Horner, D. S., Hirt, R. P., Kilvington, S., Lloyd, D., and Embley, T. M. (1996) Molecular data suggest an early aquisition of the mitochondrion endosymbiont. Proc. R. Soc. London Ser. B 263: 1053–1059.
51.Howitz, K.T., Bitterman, K.J., Cohen, H.Y., Lamming, D.W., Lavu, S., Wood, J.G., Zipkin, R.E., Chung, P., Kisielewski, A., and Zhang, L.L. (2003) Small molecule activators of sirtuins extend Saccharo- myces cerevisiae lifespan. Nature 425:191–196.
52.Hrdy, I., Mertens, E., and Van Schaftingen, E. (1993) Identification, purification and separation of different isoenzymes of NADP-specific malic enzymefrom Tritrichomonas foetus. Mol. Biochem. Parasitol. 57:253–260.
53.Hursting, S.D., Lavigne, J.A., Berrigan, D., Perkins, S.N., and Barrett, J.C. (2003) Calorie restriction, aging, and cancer prevention: mechanisms of action and applicability to humans. Annu. Rev. Med. 54:131–152.
54.Imai, S., Armstrong, C.M., Kaeberlein, M., and Guarente, L. (2000) Transcriptional silencing and longevity protein Sir2 is an NAD- dependent histone deacetylase. Nature 403:795–800.
55.Ingram, D.K., Anson, R.M., de Cabo, R., Mamczarz, J., Zhu, M., Mattison, J., Lane, M.A., and Roth, G.S. (2004). Development of calorie restriction mimetics as a prolongevity strategy. Ann. N. Y. Acad. Sci. 1019:412–423.
56.Ishitani, R. and Chuang, D.M. (1996) Glyceraldehyde-3-phosphate
dehydrogenase antisense oligodeoxynucleotides protect against cytosine arabinonucleoside-induced apoptosis in cultured cerebellar
neurons. Proc. Natl. Acad. Sci. U. S. A. 93: 9937–9941.
57.Johnson, P. J., d’Oliveira, C. E. Gorrell, T. E., and Muller M. (1990) Molecular analysis of the hydrogenosomal ferredoxin of the anaerobic protest Trichomonas vaginalis. Proc. Natl. Acad. Sci. USA 87: 6097–6101.
58.Johnson, P. J., Lahti, C. J., and Bradley, P. J. (1993) Biogenesis of the hydrogenosome in the anaerobic protist Trichomonas vaginalis. J. Parasitol. 79:664–670.
59.Kaeberlein, M., McVey, M., and Guarente, L. (1999) The SIR2/3/4 complex and SIR2 alone promote longevity in Saccharomyces cerevisiae by two different mechanisms. Genes Dev. 13: 2570–2580.
60.Kharsany, A. B., Hoosen,A. A., Moodley, J., Bagaratee, J., and Gouws, E. (1993) The association between sexually transmitted pathogens and cervical intra-epithelial neoplasia in a developing community. Genitourin. Med. 69:357–360.
61.Khoshnan, A., and John, F. A., (1993) Multiple double-stranded RNA segments are associated with virus particles infecting Trichomonas vaginalis. J. Virol. 67:6950–6955.
62.Kimura, K., Tissenbaum, H. A., Liu, Y., and Ruvkun, G. (1997) The daf-2 insulin receptor family member regulates longevity and diapause in Caenorhabditis elegans. Science 277:942–946.
63.Kim, J.W. et al. (2004) Evaluation of myc E-box phylogenetic footprints in glycolytic genes by chromatin immunoprecipitation assays. Mol.Cell. Biol. 24:5923–5936.
64.Laga, M., Nzila, N., and Goeman, J. (1991) The interrelationship of sexually transmitted diseases and HIV infection: implications for the control of both epidemics in Africa. AIDS 5(Suppl. 1):S55–S63.
65.Laga, M., Manoka, A., Kivuvu, M., Malele, B., Tuliza, M., Nzila, N., and Goeman, J. B. (1993) Non-ulcerative sexually transmitted diseases as risk factors for HIV-1 transmission in women: results from a cohort study. AIDS 7:95–102.
66.Laga, M., Alary, M., Nzila, N., Manoka, A. T., Tuliza, M., Behets, F., Goeman, J. S., and Louis, M. P. (1994) Condom promotion, sexually transmitted disease treatment, and declining incidence of HIV-1 infection in female Zairian sex workers. Lancet 344:246–248.
67.Lahti, C. J., d’Oliveira, C. E., and Johnson, P. J. (1992) Beta- succinylcoenzyme A synthetase from Trichomonas vaginalis is a soluble hydrogenosomal protein with an amino-terminal sequence that resembles mitochondrial presequences. J. Bacteriol. 174: 6822– 6830.
68.Landry, J., Sutton, A., Tafrov, S.T., Heller, R.C., Stebbins, J., Pillus, L., and Sternglanz, R. (2000) The silencing protein SIR2 and its homologs are NAD-dependent protein deacetylases. Proc. Natl. Acad. Sci. USA 97: 5807–5811.
69.Langley, E., Pearson, M., Faretta, M., Bauer, U.M., Frye, R.A., Minucci, S., Pelicci, P.G., and Kouzarides, T. (2002) Human SIR2 deacetylates p53 and antagonizes PML/p53-induced cellular sense- cence. EMBO J. 21: 2383–2396.
70.Lapan B, Friedman M. M. (1950) Glycogen and reducing substances in vaginal mucus: gestational and cyclical variations. Am J Obstet Gynecol . 1:361-376.
71.Lin, K., Dorman, J. B., Rodan, A. and Kenyon, C. (1997) daf-16: An HNF-3/forkhead family member that can function to double the life-span of Caenorhabditis elegans. Science 278:1319–1322.
72.Lin, S.J., Defossez, P.A., and Guarente, L. (2000) Requirement of NAD and SIR2 for life-span extension by calorie restriction in Saccharomyces cerevisiae. Science 289:2126–2128.
73.Luo, J., Nikolaev, A.Y., Imai, S., Chen, D., Su, F., Shiloh, A., Guarreente, L., and Gu, W. (2001). Negative control of p53 by Sir2α promotes cell survival under stress. Cell 107:137–148.
74.Lindmark, D. G., and Muller, M. (1973) Hydrogenosome, a cytoplasmic organelle of the anaerobic flagellate Tritrichomonas foetus, and its role in pyruvate metabolism. J. Biol. Chem. 248:7724–7728.
75.Lindmark, D. G., Muller, M., and Shio, H. (1975) Hydrogenosomes in Trichomonas vaginalis. J. Parasitol. 61:522–554.
76.Lindmark, D. G., Eckenrode, B. L., Halberg, L. A., and Dinbergs, I. D. (1989) Carbohydrate, energy and hydrogenosomal metabolism of Tritrichomonas foetus and Trichomonas vaginalis. J. Protozool. 36:214–216.
77.Lirosi, G., and Guarascio A. (1972) Effects of hormonal changes in the vaginal environment in the treatment of vaginitis especially due to Trichomonas. Minerva Ginecol. 24:23–27.
78.Luo, J., Nikolaev, A.Y., Imai, S., Chen, D., Su, F., Shiloh, A., Guarente, L., and Gu, W. ( 2001) Negative control of p53 by Sir2a promotes cell survival under stress. Cell 107:137–48.
79.Mack, S. R., and Muller, M. (1980) End products of carbohydrate metabolism in Trichomonas vaginalis. Comp. Biochem. Physiol. 67: 213–216.
80.Majewski, N. et al. (2004) Akt inhibits apoptosis downstream of BID
cleavage via a glucose-dependent mechanism involving mitochondrial hexokinases. Mol. Cell. Biol. 24:730–740.
81.Mallinson, D. J., Lockwood, B. C., Coombs, G. H., and North, M. J. (1994) Identification and molecular cloning of four cysteine proteinase genes from the pathogenic protozoon Trichomonas vaginalis. Microbiology 140:2725–2735.
82.Manson, P. R. (1979) Serodiagnosis of Trichomonas vaginalis infection by indirect fluorescent antibody test. J. Parasitol. 32: 1211–1215.
83.Marczak, R., Gorrell, T. E., and Muller, M. (1983) Hydrogenosomal ferredoxin of the anaerobic protozoon, Tritrichomonas foetus. J. Biol. Chem. 258:12427–12433.
84.McCay, C.M., Cromwell, M.F., and Maynard, L.A. (1935) The effect of retarded growth upon the length of life span and ultimate body size. J. Nutr. 10:63–79.
85.McLellan, R., Spence, M. Brockman, R., Raffel, M. L., and Smith, J. L., (1982) The clinical diagnosis of trichomoniasis. Obstet. Gynecol. 60:30–34.
86.Minkoff, H., Grunebaum, A. N., Schwarz, R. H., Feldman, J., Cummings, M. C., Clark, W. L., Pringle, G., and McCormack W. M. (1984) Risk factors for prematurity and premature rupture of membranes: a prospective study of the vaginal flora in pregnancy. Am. J. Obstet. Gynecol. 150:965–972.
87.Moldwin, R. M. (1992) Sexually transmitted protozoal infections. Trichomonas vaginalis, Entamoeba histolytica, and Giardia lamblia. Urol. Clin. North Am. 19:93–101.
88.Motta, M.C., Divecha, N., Lemieux, M., Kamel, C., Chen, D., Gu, W., Bultsma, Y., McBurney, M.W., and Guarente, L. (2004). Mammalian SIRT1 represses forkhead transcription factors. Cell 116: 551–563.
89.Pajvani, U.B., and Scherer, P.E. (2003). Adiponectin: systemic contributor to insulin sensitivity. Curr. Diab. Rep. 3:207–213.
90.Müller, M. (1980) The hydrogenosome. Symp. Soc. Gen. Microbiol. 30:127–142.
91.Müller, M. (1986) Reductive activation of nitroimidazoles in aerobic microorganisms. Biochem. Pharmacol. 35:37–41.
92.Müller, M. (1987) Hydrogenosomes of trichomonad flagellates. Acta Univ. Carol. Biol. 30:249–260.
93.Müller, M. (1988) Energy metabolism of protozoa without mitochondria. Annu. Rev. Microbiol. 42:465–488.
94.Müller, M. (1990) Biochemistry of Trichomonas vaginalis, p. 53–83. In B. M. Honigberg. (ed.), Trichomonads parasitic in humans. Springer-Verlag, NewYork, N.Y.
95.Müller, M. (1992) Energy metabolism of ancestral eukaryotes: hypothesis based on the biochemistry of amitochondriate parasitic protists. Chin. Med.J. 28:33–40.
96.Mullis, K., Fallona, F., Scharf, S., Saiki, R., Horn, G., and Erilich, H., (1986) Specific enzymatic amplification of DNA in vitro: the polymerase chain reaction. Cold Spring Harb Symp Quant Biol. 51: 263-73.
97.Niinaka, Y. et al. (1998) Expression and secretion of neuroleukin/ phosphohexoseisomerase/maturation factor as autocrine motility factor by tumor cells. Cancer Res. 58:2667–2674.
98.Ogg, S. et al. (1997) The Fork head transcription factor DAF-16 transduces insulin-like metabolic and longevity signals in C. elegans. Nature 389:994–999.
99.Ovcinnikov, N. M., Delektorskij, V. V., Turanova, E. N., and Yashkova, G. N. (1975) Further studies of Trichomonas vaginalis with transmission and scanning electron microscopy. Br. J. Vener. Dis. 51:357–375.
100.Pastorino, J.G. et al. (2002) Mitochondrial binding of hexokinase II inhibits Bax-induced cytochrome c release and apoptosis. J. Biol.
Chem. 277: 7610–7618.
101.Pereira-Neves, A., Riberio, K. C., Benchimol, M. (2003) Pseudocysts in trichomonads--new insights. Protist. 154: 313-29.
102.Picard, F., Kurtev, M., Chung, N., Topark-Ngarm, A., Senawong, T., Machado, R., Leid, M., McBurney, M.W., and Guarente, L. (2004). Sirt1 promotes fat mobilization in white adipocytes by reprssing PPAR-gamma. Nature 429: 771-776.
103.Rathmell, J.C. et al. (2003) Akt-directed glucose metabolism can
prevent Bax conformation change and promote growth factor independent survival. Mol. Cell. Biol. 23: 7315–7328.
104.Ray, R.B. et al. (1995) Human breast carcinoma cells transfected with the gene encoding a c-myc promoter-binding protein (MBP-1) inhibits tumors in nude mice. Cancer Res. 55:3747–3751.
105.Rein, M. F. and Chapel, T. A. (1975) Trichomoniasis, candidiasis, and the minor venereal disease. Clin. Obstet. Gynecol. 18:73-88.
106.Riley, D. E., Krieger, J. N., Rabinovitch, P. S. (1994) Trichomonas vaginalis: dominant G2 period and G2 phase arrest in a representative of an early branching eukaryotic lineage. J. Eukaryot. Microbiol. 41: 408-14.
107.Reiss, N. et al. (1986) Five enzymes of the glycolytic pathway serve as substrates for purified epidermal-growth-factor-receptor kinase. Biochem. J. 239:691–697.
108.Rodriguez, A. et al. (2001) The hexokinase 2 protein regulates the expression of the GLK1, HXK1 and HXK2 genes of Saccharomyces cerevisiae. Biochem. J. 355: 625–631
109.Rogina, B., Helfand, S. L., Frankel, S. (2002) Longevity regulation by Drosophila Rpd3 deacetylase and caloric restriction. Science 298: 1745.
110.Sebek, V. (1972) Trichomonas pyosalpinx. Cesk. Gynekol. 37:14-15. (In Czech)
111.Schena M., Shalon D., Davis R. W., and Brown P.O. (1995) Quanti- tative monitoring of gene expression patterns with a complementary DNA microarray. Science 270:467-470.
112.Sohal, R.S., and Weindruch, R. (1996) Oxidative stress, caloric restriction, and aging. Science 273:59–63.
113.Sreekumar, R., Unnikrishnan, J., Fu, A., Nygren, J., Short, K.R., Schimke, J., Barazzoni, R., and Nair, K.S. (2002) Effects of caloric restriction on mitochondrial function and gene transcripts in rat muscle. Am. J. Physiol. Endocrinol. Metab. 283:E38–E43.
114.Steinbuchel, A., and Muller, M. (1986) Glycerol, a metabolic end product of Trichomonas vaginalis and Tritrichomonas foetus. Mol. Biochem. Parasitol. 20:45–55.
115.Steinbuchel, A., and Muller, M. (1986) Anaerobic pyruvate metabolism of Tritrichomonas foetus and Trichomonas vaginalis hydrogenosomes. Mol.Biochem. Parasitol. 20:57–65.
116.Stephen, R. S. (2006) Use of microarray biomarkers to identify longevity therapeutics. Aging Cell 5:39-50.
117.Subramanian, A. and Miller, D.M. (2000) Structural analysis of alphaenolase. Mapping the functional domains involved in down-regulation of the c-myc protooncogene. J. Biol. Chem. 275: 5958–5965.
118.Sun, Y.J. et al. (1999) The crystal structure of a multifunctional
protein: phosphoglucose isomerase/autocrine motility factor/neuroleukin. Proc. Natl. Acad. Sci. U. S. A. 96, 5412–5417.
119.Tajima, H. et al. (1999) Over-expression of GAPDH induces apoptosis in COS-7 cells transfected with cloned GAPDH cDNAs. Neuroreport. 10:2029–2033.
120.ter Kuile, B. H. 1994. Carbohydrate metabolism and physiology of the parasitic protist Trichomonas vaginalis studied in chemostats. Microbiology 140:2495–2502.
121.ter Kuile, B. H. 1994. Adaptation of the carbon metabolism of Trichomonas vaginalis to the nature and availability of the carbon source. Microbiology 140:2503–2510.
122.ter Kuile, B. H., and M. Muller. 1995. Maltose utilization by extracellular hydrolysis followed by glucose transport in Trichomonas vaginalis. Parasitology 110:37–44.
123.Trumbly, R.J. (1992) Glucose repression in the yeast Saccharomyces cerevisiae. Mol. Microbiol. 6, 15–21
124.Vaziri, H., Dessain, S.K., Ng Eaton, E., Imai, S.I., Frye, R.A., Pandita, T.K., Guarente, L., and Weinberg, R.A. (2001) hSIR2(SIRT1) functions as an NAD-dependent p53 deacetylase. Cell 107:149–159.
125.Vergnes, B., Sereno, D., Madjidian-Sereno, N., Lemesre, J., and Ouaissi, A., (2002) Cytoplasmic SIR2 homologue overexpression promotes survival of Leishmania parasites by preventing programmed cell death. Gene 296:139–150.
126.Warton, A., and Honigberg B. M. (1979) Structure of trichomonads as revealed by scanning electron microscopy. J. Protozool. 26: 56–62.
127.Wellerson, R., and Kupferberg, A. B. (1962) On glycolysis in Trichomonas vaginalis. J. Protozool. 9:418–424.
128.Weindruch, R., and Walford, R.L. (1988) The Retardation of Aging and Disease by Dietary Restriction. (Springfield, Illinois: Charles C.Thomas)
129.Wisdom, A. R., and Dunlop, E. M. C. (1965) Trichomoniasis: study of the disease and its treatment in women and men. Br. J. Vener. Dis. 41:90–96.
130.Williams, K.R. et al. (1985) Identification of a nucleic acid helixdestabilizing protein from rat liver as lactate dehydrogenase-5. Proc. Natl. Acad. Sci. U. S. A. 82:5260–5264.
131.Whittington, M. J. (1957) Epidemiology of infections with Trichomonas vaginalis in the light of improved diagnostic methods. Br. J. Vener. Dis. 33:80–91.
132.Wolner-Hanssen, P., Krieger, J. N., Stevens, C. E., Kiviat, N. B., Koutsky, L., DeRouen, T., Hillier, S., and Holmes, K. K. (1989) Clinical manifestations of vaginal trichomoniasis. JAMA 261: 571–576.
133.World Health Organization. (1995) An overview of selected curable sexually transmitted diseases, p. 2–27. In Global program on AIDS. World Health Organization, Geneva, Switzerland.
134.Xiao, H., Massaro, D., Decarlo, G., and Clerch, L.B. (2004). Expression of lung uncoupling protein-2 mRNA is modulated developmentally and by caloric intake. Exp. Biol. Med. 229: 479– 485.
135.Yahiaoui, B., Taibi, A., and Ouaissi, A. (1996) A Leishmania major protein with extensive homology to silent information regulator 2 of Saccharomyces cerevisiae. Gene 169:115–118.
136.Yoon, K., Ryu, J. S., and Min, D. Y. (1991) Cytotoxicity of lymphokine activated peritoneal macrophages against Trichomonas vaginalis. Kor. J.Parasitol. 29:381–388. (In Korean.)
137.Zhang, Z. F., and Begg. C. B. (1994) Is Trichomonas vaginalis a cause of cervical neoplasia? Results from a combined analysis of 24 studies. Int. J. Epidemiol. 23:682–690.
138.Zheng, L. et al. (2003) S phase activation of the histone H2B promoter by OCA-S, a coactivator complex that contains GAPDH as a key component. Cell 114:255–266.