The mechanisms by which acidophilic bacteria such as T ferrooxidans and L ferrooxidans oxidize iron and/or sulfur is the topic of investigation by numerous researchers. In the environment, oxidation of solid mineral sulfides such as pyrite (FeS2) is greatly increased by acidophilic bacteria, and leads to production of acidic waters (known as acid mine drainage, AMD). T ferrooxidans are known to attach firmly to a solid mineral substrate such as pyrite and oxidize through direct of indirect mechanisms (Rossi, 1990), and involve accumulation of Fe 31, in the exopolymer layer, which is used as the oxidant (Blake et. al., 1992). It is believed that T ferrooxidans attach firmly to pyrite to aid in the sequestering of intermediate thiosulfate compounds, which are used as the primary energy source (Sand et al., 1995). While tremendous progress has been made on the processes involved oxidation of solid minerals such as pyrite by T ferrooxidans, and, L ferrooxidans, little has been done to show that these processes occur by these or other acidophilic organisms in nature by the same or similar mechanism. I am currently maintaining enrichment cultures of acidophilic organism, obtained by inoculating with AMD and pyritic sediment (average grain size = 100mm) from a site near Redding, CA. I would like to analyze the pyritic sediments and the microorganisms associated with them for elemental distribution by energy-filtered transmission electron microscopy (EFIEM). I am interested in using cryofixation techniques as a method for preserving the original elemental distributions within the periplasm. and the outer membrane of the organisms attached to pyrite. The reasons for this are 1) to detect the presence or absence of rusticyanin protein in the cells periplasmic space. The proteins contain copper, which should be easily detected by EFFEM; 2) to detect concentrations of iron in the outer membranes. This would allow us to confirm or disprove importance of the indirect and direct mechanisms of pyrite oxidation by these organisms. VVhile the short-term goals of this project involve cryofixation of organisms and pyrite form enrichment cultures, I would also like to apply this technique to """"""""fi-esh"""""""" samples from AMD sits. This will provide an important comparison of I)pure laboratory cultures, form which all of our current knowledge of iron and sulfur metabolism comes; 2) mixed laboratory cultures inoculated with natural sediments and water, and 3) fresh sediment sarnples from the environment. BCXZNTXFXC SUBPROJECT GRANT NUMBER: P41RR00570-27

Agency
National Institute of Health (NIH)
Institute
National Center for Research Resources (NCRR)
Type
Biotechnology Resource Grants (P41)
Project #
3P41RR000570-28
Application #
6278496
Study Section
Project Start
1998-07-01
Project End
2000-06-30
Budget Start
1997-10-01
Budget End
1998-09-30
Support Year
28
Fiscal Year
1998
Total Cost
Indirect Cost
Name
University of Wisconsin Madison
Department
Type
DUNS #
161202122
City
Madison
State
WI
Country
United States
Zip Code
53715
Malecki, Marek; Putzer, Emily; Sabo, Chelsea et al. (2014) Directed cardiomyogenesis of autologous human induced pluripotent stem cells recruited to infarcted myocardium with bioengineered antibodies. Mol Cell Ther 2:
Malecki, Marek (2014) 'Above all, do no harm': safeguarding pluripotent stem cell therapy against iatrogenic tumorigenesis. Stem Cell Res Ther 5:73
Mavroudi, Maria; Zarogoulidis, Paul; Porpodis, Konstantinos et al. (2014) Stem cells' guided gene therapy of cancer: New frontier in personalized and targeted therapy. J Cancer Res Ther (Manch) 2:22-33
Malecki, Marek; Dahlke, Jessica; Haig, Melissa et al. (2013) Eradication of Human Ovarian Cancer Cells by Transgenic Expression of Recombinant DNASE1, DNASE1L3, DNASE2, and DFFB Controlled by EGFR Promoter: Novel Strategy for Targeted Therapy of Cancer. J Genet Syndr Gene Ther 4:152
Zarogoulidis, Paul; Darwiche, Kaid; Sakkas, Antonios et al. (2013) Suicide Gene Therapy for Cancer - Current Strategies. J Genet Syndr Gene Ther 4:
Malecki, Marek; Sabo, Chelsea; Putzer, Emily et al. (2013) Recruitment and retention of human autologous CD34+ CD117+ CD133+ bone marrow stem cells to infarcted myocardium followed by directed vasculogenesis: Novel strategy for cardiac regeneration. Mol Cell Ther 1:
Malecki, Marek; LaVanne, Christine; Alhambra, Dominique et al. (2013) Safeguarding Stem Cell-Based Regenerative Therapy against Iatrogenic Cancerogenesis: Transgenic Expression of DNASE1, DNASE1L3, DNASE2, DFFB Controlled By POLA1 Promoter in Proliferating and Directed Differentiation Resisting Human Autologous Pluripotent J Stem Cell Res Ther Suppl 9:
Malecki, Marek; Tombokan, Xenia; Anderson, Mark et al. (2013) TRA-1-60(+), SSEA-4(+), POU5F1(+), SOX2(+), NANOG(+) Clones of Pluripotent Stem Cells in the Embryonal Carcinomas of the Testes. J Stem Cell Res Ther 3:
Malecki, Marek (2013) Improved targeting and enhanced retention of the human, autologous, fibroblast-derived, induced, pluripotent stem cells to the sarcomeres of the infarcted myocardium with the aid of the bioengineered, heterospecific, tetravalent antibodies. J Stem Cell Res Ther 3:
Malecki, Marek; Malecki, Bianca (2012) Routing of Biomolecules and Transgenes' Vectors in Nuclei of Oocytes. J Fertili In Vitro 2012:108-118

Showing the most recent 10 out of 24 publications