Biochemistry Commons^™

Evidence For An Epigenetic Mechanism By Which Hsp90 Acts As A Capacitor For Morphological Evolution, Vincent E. Sollars, Xiangyi Lu, Li Xiao, Xiaoyan Wang, Mark D. Garfinkel, Douglas M. Ruden

Biochemistry and Microbiology

Morphological alterations have been shown to occur in Drosophila melanogaster when function of Hsp90 (heat shock 0-kDa protein 1α, encoded by Hsp83) is compromised during development1. Genetic selection maintains the altered phenotypes in subsequent generations1. Recent experiments have shown, however, that phenotypic variation still occurs in nearly isogenic recombinant inbred strains of Arabidopsis thaliana2. Using a sensitized isogenic D. melanogaster strain, iso-KrIf-1, we confirm this finding and present evidence supporting an epigenetic mechanism for Hsp90’s capacitor function, whereby reduced activity of Hsp90 induces a heritably altered chromatin state. The altered chromatin state is evidenced by ectopic expression …

Calcium Binding To Calmodulin Mutants Monitored By Domain-Specific Intrinsic Phenylalanine And Tyrosine Fluorescence, W. S. Vanscyoc, B. R. Sorensen, E. Rusinova, William Laws, J. B. A. Ross, M. A. Shea

Chemistry and Biochemistry Faculty Publications

Cooperative calcium binding to the two homologous domains of calmodulin (CaM) induces conformational changes that regulate its association with and activation of numerous cellular target proteins. Calcium binding to the pair of high-affinity sites (III and IV in the C-domain) can be monitored by observing calcium-dependent changes in intrinsic tyrosine fluorescence intensity (lambda(ex)/lambda(em) of 277/320 nm). However, calcium binding to the low-affinity sites (I and II in the N-domain) is more difficult to measure with optical spectroscopy because that domain of CaM does not contain tryptophan or tyrosine. We recently demonstrated that calcium-dependent changes in intrinsic phenylalanine fluorescence (lambdaex/lambdaem of …

Anisomycin Activates Jnk And Sensitises Du 145 Prostate Carcinoma Cells To Fas Mediated Apoptosis, James Curtin, Thomas Cotter

Articles

Treatment of the hormone refractory prostate cancer cell line DU 145 with sublethal concentrations of chemotherapeutic drugs has been reported to sensitise these cells to Fas mediated apoptosis. However, the mechanism by which this occurs has not been determined. Our group has shown that inhibition of JNK activity completely abrogates the effects of chemotherapeutic drugs. Using anisomycin, a potent JNK agonist, we have demonstrated a role for JNK in Fas mediated apoptosis in DU 145 cells. Inhibition of Caspase 8 and Caspase 9 completely inhibits this process which suggests that DU 145 cells require mitochondrial amplification of the Fas apoptotic …

Electrically Mediated Delivery Of Vector Plasmid Dna Elicits An Antitumor Effect, L. Heller, D. Coppola

Bioelectrics Publications

In vivo electroporation is an efficient means of increasing plasmid DNA delivery to normal tissues, such as skin and muscle, as well as directly to tumors. In the experiments described here, plasmid DNA was delivered by in vivo electroporation to B16 mouse melanomas using two very different pulsing protocols. Reporter expression increased 21- or 42-fold, respectively with electroporation over injection alone. The growth of experimental melanomas with an approximate diameter of 4 mm on the day of treatment was monitored after electroporation delivery of reporter plasmid DNA. Remarkably, short-term complete regressions using one of these pulsing protocols occurred in up …

Guanine And 7,8-Dihydro-8-Oxo-Guanine-Specific Oxidation In Dna By Chromium(V), Kent D. Sugden, Brooke Martin

Chemistry and Biochemistry Faculty Publications

The hexavalent oxidation state of chromium [Cr(VI)] is a well-established human carcinogen, although the mechanism of cancer induction is currently unknown. Intracellular reduction of Cr(VI) forms Cr(V), which is thought to play a fundamental role in the mechanism of DNA damage by this carcinogen. Two separate pathways of DNA damage, an oxidative pathway and a metal-binding pathway, have been proposed to account for the lesions observed in cell systems. We have used a model Cr(V) complex, N,N´-ethylenebis(salicylidene-animato)oxochromium(V) [Cr(V)-Salen], to investigate the oxidative pathway of DNA damage and to elucidate the lesions generated from this oxidation process. Reaction of Cr(V)-Salen with …

Monte Carlo Simulation On The Indirect Exchange Interactions Of Co-Doped Zno Film, Abraham F. Jalbout, Hanning Chen, Scott L. Whittenburg

Chemistry and Biochemistry Faculty Publications

Monte Carlo simulations using a three-dimensional lattice model studied the Ruderman–Kittel–Kasuya–Yosida (RKKY) indirect exchange interaction of doped magnetic Co ions in ZnOfilms. The results of the calculations show that the RKKY interaction in Co-doped ZnO is long ranged and its magnitude is proportional to (inverse of the distance from a central ion). The sign oscillates with a frequency that depends on the concentration of the carrier. The long-distance sum of the RKKY indirect exchange energies is positive indicating that these materials are ferromagnetic, in direct correlation with previously reported results.

The Atp:Co(I)Rrinoid Adenosyltransferase (Coba) Enzyme Of Salmonella Enterica Requires The 2'-Oh Group Of Atp For Function And Yields Inorganic Triphosphate As Its Reaction Byproduct, Maris V. Fonseca, Nicole R. Buan, Alexander R. Horswill, Ivan Rayment, Jorge C. Escalante-Semerena

Department of Biochemistry: Faculty Publications

The specificity of the ATP:corrinoid adenosyltransferase (CobA) enzyme of Salmonella enterica serovar Typhimurium LT2 for its nucleotide substrate was tested using ATP analogs and alternative nucleotide donors. The enzyme showed broad specificity for the nucleotide base and required the 2’-OH group of the ribosyl moiety of ATP for activity. ³¹P NMR spectroscopy was used to identify inorganic triphosphate (PPP_i) as the byproduct of the reaction catalyzed by the CobA enzyme. Cleavage of triphosphate into pyrophosphate and orthophosphate did not occur, indicating that triphosphate cleavage was not required for release of the adenosylcorrinoid product. Triphosphate was a strong …

Glutathione Levels And Bax Activation During Apoptosis Due To Oxidative Stress In Cells Expressing Wild-Type And Mutant Cystic Fibrosis Transmembrane Conductance Regulator, Thomas Jungas, Iris Motta, Francis Duffieux, Pascale Fanen, Veronique Stoven, David M. Ojcius

All Dugoni School of Dentistry Faculty Articles

Cystic fibrosis is characterized by chronic inflammation and an imbalance in the concentrations of alveolar and lung oxidants and antioxidants, which result in cell damage. Modifications in lung glutathione concentrations are recognized as a salient feature of inflammatory lung diseases such as cystic fibrosis, and glutathione plays a major role in protection against oxidative stress and is important in modulation of apoptosis. The cystic fibrosis transmembrane conductance regulator (CFTR) is permeable to Cl−, larger organic ions, and reduced and oxidized forms of glutathione, and the ΔF508 CFTR mutation found in cystic fibrosis patients has been correlated with impaired glutathione transport …

Comparison Of Water Quality, Zooplankton Density, And Cover In Razorback Sucker (Xyrauchen Texanus [Abbott]) Spawning Areas Of Lake Mead And Lake Mohave, Michael E. Golden, Paul B. Holden, Southern Nevada Water Authority

Publications (WR)

Las Vegas Bay and Echo Bay in Lake Mead have small, self-sustaining populations of razorback sucker (Xyrauchen texanus [Abbot]). Increased productivity and cover have been hypothesized as reasons for successful recruitment of razorback sucker in Lake Mead. Conversely, reproduction has been documented on Lake Mohave, another lower Colorado River reservoir, but no recruitment has been observed. In 2000, BIO-WEST, Inc. was contracted by the Southern Nevada Water Authority to design and implement a study to examine nutrient levels, zooplankton density, and cover in areas with and without razorback sucker recruitment success. We sampled Echo Bay, Las Vegas Bay, and …

Regulation And Measurement Of Oxidative Stress In Apoptosis, James Curtin, Maryanne Donovan, Thomas Cotter

Articles

Cells are constantly generating reactive oxygen species (ROS) during aerobic metabolism. As a consequence, each cell is equipped with an extensive antioxidant defence system to combat excessive production of ROS. Oxidative stress occurs in cells when the generation of ROS overwhelms the cell's natural antioxidant defences. There is a growing consensus that oxidative stress and the redox state of a cell plays a pivotal role in regulating apoptosis, a tightly controlled form of cell death in which a cell partakes in its own demise. More recently, a role for reactive nitrogen species (RNI) as both positive and negative regulators of …

Magnetization Reversal Of Elliptical Co/Cu/Co Pseudo-Spin Valve Dots, Ngocnga Dao, Scott L. Whittenburg, Y. Hao, Leszek M. Malkinski, Jian Qing Wang, C. A. Ross

Chemistry and Biochemistry Faculty Publications

We present our recent simulated results on Cr (5 nm)/ Cu (5 nm)/ Co (5 nm)/ Cu (3 nm)/ Co (2 nm) pseudo-spin valve dots. The simulated results agree qualitatively with the experimental results. Three different sizes of elliptical dots, and were simulated. Our simulations show that in these types of dots magnetization reversal occurs by the formation of domain walls: for and for No domain wall was observed in the reversal of the dots. For such dots, the simulated loops show a small two-step reversal pattern with the thin upper layer partially reversing followed by complete reversal of both …

Inhibition Of Apoptosis By Gamma Interferon In Cells And Mice Infected With Chlamydia Muridarum (The Mouse Pneumonitis Strain Of Chlamydia Trachomatis), Jean-Luc Perfettini, Toni Darville, Alice Dautry-Varsat, Roger G. Rank, David M. Ojcius

All Dugoni School of Dentistry Faculty Articles

The effect of gamma interferon (IFN-γ) on apoptosis due to infection by Chlamydia muridarum (the mouse pneumonitis strain of Chlamydia trachomatis) was studied in epithelial cells in culture and in the genital tracts of mice. IFN-γ concentrations that induce the formation of aberrant, persistent chlamydiae inhibit apoptosis due to C. muridarum infection. In cells treated with an IFN-γ concentration that leads to the development of a heterogenous population of normal and aberrant Chlamydia vacuoles, apoptosis was inhibited preferentially in cells that contained the aberrant vacuoles. The inhibitory effect of IFN-γ appears to be due in part to expression of host …

Vectorial Acylation In Saccharomyces Cerevisiae, Zhiying Zou, Fumin Tong, Nils J. Færgeman, Claus Børsting, Paul N. Black, Concetta Dirusso

Department of Biochemistry: Faculty Publications

In Saccharomyces cerevisiae Fat1p and fatty acyl-CoA synthetase (FACS) are hypothesized to couple import and activation of exogenous fatty acids by a process called vectorial acylation. Molecular genetic and biochemical studies were used to define further the functional and physical interactions between these proteins. Multicopy extragenic suppressors were selected in strains carrying deletions in FAA1 and FAA4 or FAA1 and FAT1. Each strain is unable to grow under synthetic lethal conditions when exogenous long-chain fatty acids are required, and neither strain accumulates the fluorescent long-chain fatty acid C₁-BODIPY-C₁₂ indicating a fatty acid transport defect. By using these …

Serotyping Group B Streptococci In A Small Community Hospital: An Analysis Of Distribution And Site Of Isolation, Jennifer M. Smith, Jason A. Rexroth, David G. Chaffin, Susan H. Jackman

Biochemistry and Microbiology

Objective: To determine the prevalence and site of isolation of different serotypes of group B streptococcus (GBS) colonization or infection at a small community hospital.

Methods: GBS isolates were obtained from a small community hospital and were then serotyped as la, Ib, II, III, IV, V or non-typeable. Hospital records were reviewed for patient sex, age and pregnancy status as well as the site of GBS isolation.

Results: GBS serotypes Ia, III and V were most common and accounted for over 60% of the total number of isolates. Serotype Ia was most prevalent in reproductive-age females, while serotypes V and …

Murine Epidermal Cell Antigen (Skn)-Directed Autoimmunity Induced By Transfer Of Cd4+ T Cells, Susan H. Jackman, Shivaleela Keerthy, Giselle Perry

Biochemistry and Microbiology

While pathogenic T cells have been identified for several diseases with epithelial cell damage, an autoimmune T cell-mediated response targeted against a known keratinocyte antigen has not been reported. Previously we described an autoimmune response directed to the mouse epidermal cell antigens, Skn. For our murine model, primed Skn-immune lymphocytes are adoptively transferred to recipients, which develop lesions at the site of mild skin trauma. In this study we investigated the nature of the autoimmune component of the Skn response. A time-course study demonstrated a relationship between the number of primed Sknimmune cells injected and the severity of skin lesions …

The Southern African Regional Science Initiative (Safari 2000): Overview Of The Dry Season Field Campaign, R. J. Swap, H. J. Annegarn, J. T. Suttles, J. Haywood, M. C. Helmlinger, C. Hely, Peter V. Hobbs, B. N. Holben, J. Ji, M. D. King, T. Landmann, W. Maenhaut, L. Otter, B. Pak, S. J. Piketh, S. Platnick, J. Privette, D. Roy, A. M. Thompson, Darold E. Ward, Robert J. Yokelson

Chemistry and Biochemistry Faculty Publications

The Southern African Regional Science Initiative (SAFARI 2000) is an international science project investigating the earth-atmosphere-human system in southern Africa. The programme was conducted over a two-year period from March 1999 to March 2001. The dry season field campaign (August-September 2000) was the most intensive activity and involved over 200 scientists from eighteen countries. The main objectives were to characterize and quantify biogenic, pyrogenic and anthropogenic aerosol and trace gas emissions and their transport and transformations in the atmosphere, and to validate NASA's Earth Observing System's satellite Terra within a scientific context. Five aircraft - two South African Weather Service …

Calorimetric Study Of Block-Copolymers Of Poly(N-Butyl Acrylate) And Gradient Poly(N-Butyl Acrylate-Co-Methyl Methacrylate), A I. Buzin, M Pyda, P Costanzo, K Matyjaszewski, B Wunderlich

Chemistry and Biochemistry

The nanophase separation in diblock and triblock copolymers consisting of immiscible poly(n-butyl acrylate) (block A) and gradient copolymers of methyl methacrylate (MMA) and n-butyl acrylate (n BA) (block M/A) were investigated by means of their heat capacity, C_p, as a function of the composition of the blocks M/A and temperature. In all copolymers studied, both blocks are represented by their C_p and glass transition temperature, T_g, as well as the broadening of the transition-temperature range. The low-temperature transition of the blocks A is always close to that of the …

Flexibility Of Biv Tar-Tat: Models Of Peptide Binding, M Hsieh, E D. Collins, T Blomquist, Brooke Lustig

Faculty Publications, Chemistry

No abstract provided.

Preparation Of 6-Fluoro-8-Quinolinol And Related 6-Fluoroquinolines / Herman Gershon, Donald D. Clarke, And Muriel Gershon Department Of Chemistry, Fordham University, Bronx, New York 10458 U.S.A., New York Botanical Garden, Bronx, New York 10458, U.S.A., Herman Gershon, Donald Dudley Clarke Phd, Muriel Gershon

Chemistry Faculty Publications

6-Fluoro-8-quinolinol was prepared from 2-amino-5-fluorophenol by a Skraup synthesis. No synergism was observed between 5-fluoro- and 6-fluoro-8-quinolinols or between 6-fluoro-8-quinolinolsuoro- and 7-fluoro-8-quinolinols against any of the six fungi in our test system (Aspergillus niger, A. oryzae, Myrothecium verrucaria, Trichoderma viride, Mucor cirinelloides, and Trichophyton mentagrophytes) in Sabouraud dextrose broth. Unlike the fluoro-8-quinolinols, the 8-quinolinols comparably substituted with chlorine or bromine did form synergistic mixtures. This is attributed to steric factors

Familial Als-Associated Mutations Decrease The Thermal Stability Of Distinctly Metallated Species Of Human Copper/Zinc Superoxide Dismutase, Daryl K. Eggers, J. A. Rodriguez, J. S. Valentine, J. A. Roe, A. Tiwari, R. H. Brown, L. J. Hayward

Faculty Publications, Chemistry

We report the thermal stability of wild type (WT) and 14 different variants of human copper/zinc superoxide dismutase (SOD1) associated with familial amyotrophic lateral sclerosis (FALS). Multiple endothermic unfolding transitions were observed by differential scanning calorimetry for partially metallated SOD1 enzymes isolated from a baculovirus system. We correlated the metal ion contents of SOD1 variants with the occurrence of distinct melting transitions. Altered thermal stability upon reduction of copper with dithionite identified transitions resulting from the unfolding of copper-containing SOD1 species. We demonstrated that copper or zinc binding to a subset of “WT-like” FALS mutants (A4V, L38V, G41S, G72S, D76Y, …

Fluorescence In Situ Hybridization On Sperm Using Alkaline Denaturation, L. J. Aridgides, M. Stacey, L. Brihn, D. Scott, C. Osgood

Bioelectrics Publications

Fluorescence in situ hybridization (FISH) permits the detection of unique DNA sequences of target DNA within single cells. FISH allows the identifica­tion of DNA sequences within individ­ual cells and readily identifies genetic changes within a mixed population of cells. However, problems arise in main­taining nuclear morphology, particular­ly following harsh denaturation proce­dures where cells are heated to 80°C, causing the nuclear boundary to be­come blurred or the cell to completely burst. In an attempt to preserve cellular morphology and high hybridization ef­ficiency, particularly in fresh tissue samples, we evaluated a FISH protocol using alkaline denaturation in place of heat.

Preparation And Antifungal Activity Of 3-Iodo- And 6-Iodo-8-Quinolinols / Herman Gershon, Donald D. Clarke, John J. Mcmahon, And Muriel Gershon Department Of Chemistry, Fordham University, Bronx, Ny 10458-9993, Usa New York Botanical Garden, Bronx, Ny 10458-9993, Usa, Herman Gershon, Donald Dudley Clarke Phd, John J. Mcmahon, Muriel Gershon

Chemistry Faculty Publications

3-Iodo- and 6-Iodo-8-quinolinols were prepared and tested against six fungi: Aspergillus niger, A. oryzae, Myrothecium verrucaria, Trichoderma viride, Mucor cirinelloides, and Trichophyton mentagrophytes in Sabouraud dextrose broth. A comparison with the previously known 5-iodo- and 7-iodo-8-quinolinols showed that the 6-iodo isomer was the most active

Role Of Bcl-2 Family Members In Caspase-Independent Apoptosis Due To Chlamydia Infection, Jean-Luc Perfettini, John C. Reed, Nicole Israel, Jean-Claude Martinou, Alice Dautry-Varsat, David M. Ojcius

All Dugoni School of Dentistry Faculty Articles

Infection with an obligate intracellular bacterium, the Chlamydia trachomatis lymphogranuloma venereum (LGV/L2) strain or the guinea pig inclusion conjunctivitis serovar of Chlamydia psittaci, leads to apoptosis of host cells. The apoptosis is not affected by a broad-spectrum caspase inhibitor, and caspase-3 is not activated in infected cells, suggesting that apoptosis mediated by these two strains of Chlamydia is independent of known caspases. Overexpression of the proapoptotic Bcl-2 family member, Bax, was previously shown to induce caspase-independent apoptosis, and we find that Bax is activated and translocates from the cytosol to the mitochondria in C. psittaci-infected cells. C. psittaci-induced apoptosis is …

The Atp:Co(I)Rrinoid Adenosyltransferase (Coba) Enzyme Of Salmonella Enterica Requires The 2’-Oh Group Of Atp For Function And Yields Inorganic Triphosphate As Its Reaction Byproduct, Maris V. Fonseca, Nicole R. Baun, Alexander R. Horswill, Ivan Rayment, Jorge C. Escalante-Semerena

Department of Biochemistry: Faculty Publications

The specificity of the ATP:corrinoid adenosyltransferase

(CobA) enzyme of Salmonella enterica serovar

Typhimurium LT2 for its nucleotide substrate was

tested using ATP analogs and alternative nucleotide donors.

The enzyme showed broad specificity for the nucleotide

base and required the 2’-OH group of the ribosyl

moiety of ATP for activity. ³¹P NMR spectroscopy

was used to identify inorganic triphosphate (PPP_i) as

the byproduct of the reaction catalyzed by the CobA

enzyme. Cleavage of triphosphate into pyrophosphate

and orthophosphate did not occur, indicating that

triphosphate cleavage was not required for release of

the adenosylcorrinoid product. Triphosphate was …

The Effect Of Cytosine Methylation On The Structure And Geometry Of The Holliday Junction: The Structure Of D(Ccggtacm5 Cgg) At 1.5 Å Resolution, Jeff Vargason, P. Shing Ho

Faculty Publications - Department of Biological & Molecular Science

No abstract provided.

Impact Of Sea Grass Density On Carbonate Dissolution In Bahamian Sediments, David J. Burdige, Richard C. Zimmerman

OES Faculty Publications

Carbonate dissolution has been widely observed in shallow water tropical sediments. However, sediment budgets C have generally not been closed with respect to the amount of acid required to produce the observed carbonate dissolution. Recently it has been suggested that enhanced oxygen transport into sediments through the roots and rhizomes of sea grasses might play a role in resolving this mass balance problem. We conducted studies of sea grass-carbonate sediment interactions around Lee Stocking Island, Exuma Islands, Bahamas to further examine this problem. Our studies showed that alkalinity, total dissolved inorganic carbon (ΣCO₂) and Ca²⁺ increased with …

Glass Capillary Tube Feeding: A Method For Infecting Nymphal Ixodes Scapularis (Acari: Ixodidae) With The Lyme Disease Spirochete Borrelia Burgdorferi, Anne H. Broadwater, Daniel E. Sonenshine, Wayne L. Hynes, Shane Ceraul, Aravinda M. De Silva

Biological Sciences Faculty Publications

We evaluated an artificial capillary feeding method to infect nymphal Ixodes scapularis (Say) ticks with Borrelia burgdorferi, the causative agent of Lyme disease. Thirty to 70% of the nymphs were infected after feeding for 2.5 h from glass capillary tubes filled with a solution of spirochetes. Capillary infection was stable and persisted in the nymphs for at least 10 d after feeding. Capillary feeding also maintained natural vector competence patterns because I. scapularis ticks acquired infection unlike Dermacentor variablis (Say), which did not become infected. Capillary infected I. scapularis nymphs were capable of transmitting the infection to naive mice …