Jayasimha Rao - Academia.edu (original) (raw)

Papers by Jayasimha Rao

Research paper thumbnail of Catheter-Based Medical Device Biofilm Ablation Using Histotripsy: A Parameter Study

Ultrasound in Medicine & Biology

Research paper thumbnail of Bacterial infection and endotoxin in female reproductive tract in rats: correlation with the developmental status of preimplantation embryos

Indian journal of experimental biology, 1998

During mammalian preimplantation development, a substantial numbers of embryos are believed to be... more During mammalian preimplantation development, a substantial numbers of embryos are believed to be lost for reasons that are unclear. Using female rats, we investigated whether the developmental status of embryos is influenced by bacterial infection and endotoxin in the reproductive tract. From the vagina of cycling rats (n = 11), 21 bacterial isolates were identified; they were Streptococcus faecalis (S. faecalis; 38%), Escherichia coli (E. coli; 19%), Acinetobactor calcoaceticus (A. calcoaceticus; 14%), and coagulase negative staphylococcus (14%), Micrococcus sp. (5%), Bacillus subtilis (B. subtilis; 5%) and Proteus vulgaris (P. vulgaris; 5%). From the vagina of day 4 pregnant rats (n = 12), 26 isolates were identified; they were S. faecalis (23%), A. calcoaceticus (23%), E. coli (15%), Micrococcus sp. (15%), B. subtilis (8%), P. vulgaris (4%), Staphylococcus aureus (4%), beta-hemolytic streptococcus (4%) and Pseudomonas aeruginosa (4%). Gram negative bacteria found in the vagina o...

Research paper thumbnail of A Novel Oxidized Low-Density Lipoprotein-Binding Protein, Asp-Hemolysin, Recognizes Lysophosphatidylcholine

Biological & Pharmaceutical Bulletin, 2002

Research paper thumbnail of Purification and characterization of esterases D-1 and D-2 from human erythrocytes

Archives of Biochemistry and Biophysics, 1988

Esterase D-1 (carboxylesterase; carboxylic-ester hydrolase, EC 3.1.1.1) was purified to homogenei... more Esterase D-1 (carboxylesterase; carboxylic-ester hydrolase, EC 3.1.1.1) was purified to homogeneity and esterase D-2 was highly purified from human erythrocytes. A new procedure, which included fractionation with ammonium sulfate, hydrophobic chromatography on a Toyopearl HW-65 column, and chromatographies on CM-cellulose and hydroxylapatite columns, was developed. Esterases D-1 and D-2 were purified about 9000- and 5600-fold over the precipitates with 65% saturated ammonium sulfate in 14 and 35% yields, respectively. The minimum molecular weights of esterases D-1 and D-2 were estimated to be 35,000 based on the mobilities on sodium dodecyl sulfate-polyacrylamide gel electrophoresis with or without 2-mercaptoethanol. The molecular weights of both enzymes were calculated to be 76,000 by gel filtration. These findings indicated that these two enzymes consisted of dimer without an intermolecular disulfide bond(s). Amino acid analysis of esterase D-1 showed that the total residues of aspartic acid plus asparagine, glutamic acid plus glutamine, glycine, and leucine represent about 40% of the total amino acid residues. Esterases D-1 and D-2 have almost identical biochemical characteristics, including Km values, sensitivities to sulfhydryl reagents, and molecular weights. Esterase D-2 cross-reacted with a rabbit antibody raised against the purified esterase D-1.

Research paper thumbnail of Rapid diagnosis of typhoid fever by a new ELISA test with porin antigens

Indian Journal of Medical Microbiology, 1993

... 1993 : 11(3), 191-196 RAPID DIAGNOSIS OF TYPHOID FEVER BY A NEW ELISA TEST WITH PORIN ANTIGEN... more ... 1993 : 11(3), 191-196 RAPID DIAGNOSIS OF TYPHOID FEVER BY A NEW ELISA TEST WITH PORIN ANTIGENS Jayasimha Rao*, M. Raghunath** and Nandan Singh ... MKJ Sarma & KM Nair of NIN for the useful discussions, and valuable help in the development of this method. ...

Research paper thumbnail of Novel Phosphorylcholine‐Containing Protein ofPseudomonas aeruginosaChronic Infection Isolates Interacts with Airway Epithelial Cells

The Journal of Infectious Diseases, 2008

Pseudomonas aeruginosa undergoes phase variation in the expression of the phosphorylcholine (ChoP... more Pseudomonas aeruginosa undergoes phase variation in the expression of the phosphorylcholine (ChoP) epitope, a structure crucial for the virulence of several respiratory pathogens. In this study, ChoP expression analysis comparing organisms from acute and chronic infections revealed that expression of ChoP at 37°C was higher among strains from chronic infections. Coimmunoprecipitation experiments and mass spectrometry analysis demonstrated that ChoP was on the protein elongation factor Tu. The presence of ChoP at the surface was confirmed by immunofluorescence and flow cytometry analysis of intact bacteria. Pretreatment of bronchial epithelial cells or mice with a platelet-activating factor receptor (PAFR) antagonist reduced adhesion and invasion of the ChoP-positive P. aeruginosa isolates. Results of this study suggest that ChoP expression may represent a novel phenotype expressed by the chronic infection isolates that could mediate P. aeruginosa colonization of the epithelial airway by means of the interaction with the PAFR.

Research paper thumbnail of RahU: An inducible and functionally pleiotropic protein in Pseudomonas aeruginosa modulates innate immunity and inflammation in host cells

Cellular Immunology, 2011

The aim of this study was to define the functional role of a recently identified RahU protein fro... more The aim of this study was to define the functional role of a recently identified RahU protein from Pseudomonas aeruginosa in macrophages and its role in bacterial defense. Recombinant (r)-RahU had no significant effect on cell apoptosis or cell viability in human monocytic THP-1 cells. Gene expression array of murine macrophage cells (RAW 264.7) stimulated with LPS showed modulation of common transcripts (by r-RahU and predisone) involved in inflammation. Functional cellular analysis showed RAW cells incubated with r-RahU at 1.0-10 μg/ml (0.06-0.6 μM) inhibited accumulation of nitric oxide (NO) in the presence of LPS by 10-50%. The IC(50) of r-RahU (0.6 μM) was distinct from the known inhibitors of NO production: prednisone (50 μM) and L-NMMA (100 μM). r-RahU also significantly inhibited chemotactic activity of THP-1 cells toward CCL2 or chemotactic supernatants from apoptotic T-cells. These reports show previously unknown pleiotropic properties of RahU in modulating both microbial physiology and host innate immunity.

Research paper thumbnail of Comparisons of Two Proteomic Analyses of Non-Mucoid and Mucoid Pseudomonas aeruginosa Clinical Isolates from a Cystic Fibrosis Patient

Frontiers in Microbiology, 2011

Research paper thumbnail of Prospects of riboflavin carrier protein (RCP) as an antifertility vaccine in male and female mammals

Human Reproduction Update, 1997

Introduction 325 RCP in the chicken oocyte 326 Evolutionary conservation 327 Structural modificat... more Introduction 325 RCP in the chicken oocyte 326 Evolutionary conservation 327 Structural modification of RCP for eliciting bioeffective antibodies 327 Effects of active immunization with SDS-RCM-RCP on reproductive performance of female Bonnet monkeys 328 Potential stage of immunointerference with pregnancy 328 Evaluation of C-terminal peptide of RCP as an immunogen 329 Evaluation of an N-terminal peptide (NTP) of RCP as the immunogen 330 RCP in male reproduction 330 Conclusions and future prospects 332 Acknowledgements 333 References 333 Riboflavin carrier protein (RCP) is obligatorily involved in yolk deposition of the vitamin, riboflavin, in the developing oocyte of the hen. The production of this protein is inducible by oestrogen. It is evolutionarily conserved in terms of its physicochemical, immunological and functional characteristics. It is the prime mediator of vitamin supply to the developing fetus in mammals, including primates. Passive immunoneutralization of the protein terminates pregnancy in rats. Active immunization of rats and bonnet monkeys with avian RCP prevents pregnancy without causing any adverse physiological effects of the mother in terms of her vitamin status, reproductive cycles or reproductive-endocrine profile. Denatured, linearized RCP is more effective in eliciting neutralizing antibodies capable of interfering with embryonic viability either before or during peri-implantation stages. Two defined stretches of sequential epitopes, one located at the N-terminus and the other at the C-terminus of the protein have been identified. Active immunization with either of these epitopes conjugated with diphtheria toxoid curtails pregnancy in rats and monkeys. Immunohistochemical localization of RCP on ovulated oocytes and early embryos shows that the antibodies cause degeneration only of early embryos. RCP is produced intra-testicularly and becomes localized on acrosomal surface of mammalian spermatozoa. Active immunization of male rats and monkeys with denatured RCP markedly reduces fertility by impairing the fertilizing potential of spermatozoa. These findings suggest that RCP, or its defined fragments, could be a novel, first generation vaccine for regulating fertility in both the sexes.

Research paper thumbnail of Host derived inflammatory phospholipids regulate rahU (PA0122) Gene, Protein, and Biofilm Formation in Pseudomonas aeruginosa

Cellular Immunology, 2011

Research paper thumbnail of Outer Dense Fiber Proteins Are Dominant Postobstruction Autoantigens in Adult Lewis Rats 1

Obstruction of the male reproductive tract commonly results in generation of antisperm autoantibo... more Obstruction of the male reproductive tract commonly results in generation of antisperm autoantibodies. However, only a few of the sperm autoantigens recognized by these antibodies have been characterized. To identify postobstruction rat sperm autoantigens, sperm proteins were separated by two-dimensional (2-D) gel electrophoresis. Spots corresponding to proteins that were stained by at least 50 % of postvasectomy rat sera on 2-D Western blots were removed from polyacrylamide gels and microsequenced by tandem mass spectrometry. From a total of 21 spots, 12 contained peptides that matched solely to either of two outer dense fiber proteins, odf1 or odf2. Six additional spots contained peptides comprising odf1 or odf2 and were accompanied by peptides representing other proteins. Only three spots lacked outer dense fiber peptides but did contain sequences of other known proteins. The results indicate that the outer dense fiber proteins odf1 and odf2 are dominant postobstruction autoantig...

Research paper thumbnail of Counter immuno electrophoresis of antibodies to porins (S.typhi) in serodiagnosis of typhoid fever

Indian Journal of Medical Microbiology, 1996

ABSTRACT The counter immunoelectrophoresis (CIEP) was applied to screen blood samples collected a... more ABSTRACT The counter immunoelectrophoresis (CIEP) was applied to screen blood samples collected at a single time point during second week (10-15 days) of fever from : 86 patients with typhoid fever (widal positive), five patients who were positive by blood culture for S.typhi, 27 other pyrexial cases (widal negative) and 29 healthy controls (widal negative). The percentage of widal and blood culture positive samples that were 63.7 percent positive for antiporin antibodies by the present CIEP method was found to be significantly higher (p<0.001) than that in healthy controls and other widal negative pyrexial cases. Based on these observations the characteristics of the CIEP method were computed and were found to show: sensitivity 63.7 percent, specificity 96.4 percent, accuracy 76.2 percent, positive predictive value 96.6 percent, and negative predictive value 62.0 percent respectively. None of the samples were positive for porins in serum by this method. The results suggest the usefulness of CIEP for antibodies to porins, as a simple rapid method for serodiagnosis of typhoid fever. However, the method does not seem to be useful for detection of porin antigen in early diagnosis of typhoid fever.

Research paper thumbnail of Complete Genome Sequence of Pseudomonas aeruginosa CMC-097, Isolated from a Ventilator-Associated Pneumonia Patient, Containing a Novel Carbapenem Resistance Class 1 Integron

Microbiology Resource Announcements, 2021

We report the complete genome of a clinical strain of Pseudomonas aeruginosa, CMC-097, which was ... more We report the complete genome of a clinical strain of Pseudomonas aeruginosa, CMC-097, which was isolated from a ventilator-associated pneumonia patient with a chronic infection. Illumina sequence reads were assembled using Geneious to yield a 7,044,064-bp circular chromosome containing a carbapenem resistance integron, In2020. ABSTRACT We report the complete genome of a clinical strain of Pseudomonas aeruginosa CMC-097, which was isolated from a ventilator-associated pneumonia patient with a chronic infection. Illumina sequence reads were assembled using Geneious to yield a 7,044,064-bp circular chromosome containing a carbapenem resistance integron, In2020.

Research paper thumbnail of Complete Genome Sequence of Pseudomonas aeruginosa CMC-115, a Clinical Strain from an Acute Ventilator-Associated Pneumonia Patient

Microbiology Resource Announcements, 2020

We report the complete genome of clinical strain Pseudomonas aeruginosa CMC-115, which was isolat... more We report the complete genome of clinical strain Pseudomonas aeruginosa CMC-115, which was isolated from an acute ventilator-associated pneumonia patient. Illumina sequencing reads were assembled using Geneious to yield a 6,375,262-bp circular chromosome that exhibited an unusual ferrichrome receptor in the pyoverdine synthesis locus and the absence of type 3 secretion system genes. ABSTRACT We report the complete genome of clinical strain Pseudomonas aeruginosa CMC-115, which was isolated from an acute ventilator-associated pneumonia patient. Illumina sequencing reads were assembled using Geneious to yield a 6,375,262-bp circular chromosome that exhibited an unusual ferrichrome receptor in the pyoverdine synthesis locus and the absence of type 3 secretion system genes.

Research paper thumbnail of Multidrug-resistant acinetobacter baumannii – plasmid-borne carbapenem and aminoglycoside co-resistance causing outbreak in Southwest Virginia

Research paper thumbnail of 1009. Biofilm Formation in Acinetobacter Baumannii Clinical Isolates

Open Forum Infectious Diseases

Background Multidrug resistant Acinetobacter baumannii (MDR-Ab) is a Gram-negative bacterium know... more Background Multidrug resistant Acinetobacter baumannii (MDR-Ab) is a Gram-negative bacterium known for causing severe nosocomial infections, attributed in part to its formation of biofilm. Siderophore is a virulence factor known to support biofilm formation by regulating iron availability. In this study, we screened 44 isolates of MDR-Ab from our Gram-negative repository to determine the strains that phenotypically form biofilm and produce siderophore. The results were compared to Pseudomonas aeruginosa PAO1, which produces both biofilm and siderophore. Methods Isolates were grown overnight in minimal M9 medium supplemented with casamino acids and hydroxyquinones at 37°C. Bacterial cells were normalized (to OD 600=0.01) and a standard diluted 10-3 tube was used in the study. A 96-well plate was inoculated with 100 microliters of each isolate in quadruplicates. This process was repeated in Tygon tubes with 50 microliters of each isolate in triplicates. The plate and Tygon tubes were ...

Research paper thumbnail of Focused Ultrasound Biofilm Ablation: Investigation of Histotripsy for the Treatment of Catheter-Associated Urinary Tract Infections (CAUTIs)

IEEE Transactions on Ultrasonics, Ferroelectrics, and Frequency Control

Urinary catheters often become contaminated with biofilms, resulting in catheter-associated urina... more Urinary catheters often become contaminated with biofilms, resulting in catheter-associated urinary tract infections (CAUTIs) that adversely affect patient outcomes. Histotripsy is a noninvasive focused ultrasound therapy previously developed for the noninvasive ablation of cancerous tumors and soft tissues. Histotripsy has also previously shown the ability to treat biofilms on glass slides and surgical meshes. Here, we investigate the potential of histotripsy for the treatment of CAUTIs for the first time in vitro. Clinically relevant catheter materials (Tygon, Silicone, and latex catheter mimics) and commonly used clinical catheters were tested to determine the feasibility of producing luminal histotripsy bubble clouds. A Pseudomonas aeruginosa (strain PA14) biofilm model was developed and tested to produce luminal biofilms in an in vitro Tygon catheter mimic. This model was treated with histotripsy to determine the ability to remove a luminal biofilm. Finally, the bactericidal effects of histotripsy were tested by treating PA14 suspended inside the Tygon catheter mimic. Results showed that histotripsy produced precise luminal cavitation within all tested catheter mimics and clinical catheters. Histotripsy treatment of a PA14 biofilm with histotripsy reduced luminal biofilm OD590 signal down to background levels. Further, the treatment of suspended PA14 in Manuscript

Research paper thumbnail of 2606. A Divergent Ferrichrome Receptor Associated with an Insertional Element (IS3) Identified on Novel Locus in Clinical Strain of Pseudomonas aeruginosa

Open Forum Infectious Diseases

Background Pseudomonas aeruginosa is an aerobic Gram-negative bacterium that causes life-threaten... more Background Pseudomonas aeruginosa is an aerobic Gram-negative bacterium that causes life-threatening acute and chronic infections in humans. Genotypic mutations and phenotypic variations are key features of its antimicrobial resistance and adaptation to the host environment. Pyoverdine associated genes and divergent receptors play a key role in acute Pseudomonas infections. This study seeks to address the heterogeneity of ferrichrome-iron receptor (fpvA) expression, its effect on pathogenicity and its propensity to cause acute infections clinically. Genetic and phenotypic variation of a clinical isolates of P. aeruginosa (PA097 and PA115) were identified by complete genome sequencing method. Methods An IRB-approved prospective study collected 38 P. aeruginosa clinical isolates and stored at Carilion Medical Center. Two genetically unrelated clinical strains were selected from tracheal aspirates: PA097 and PA115. These isolates were characterized by pyoverdine (pvd) quantification in...

Research paper thumbnail of 2603. Biofilm Formation as a Predictive Marker of Prognosis for Escherichia coli Sepsis

Open Forum Infectious Diseases

Background Escherichia coli is the Gram-negative organism most commonly associated with bloodstre... more Background Escherichia coli is the Gram-negative organism most commonly associated with bloodstream infections and death due to sepsis. Timely administration of appropriate antibiotic(s) plays a significant role in improving patient outcomes. E. coli expresses virulence factors (VFs) such as biofilm formation and motility phenotypes which play a role in bacterial attachment and dissemination by enabling immune system evasion and host migration. The role of these VFs in bacteremia prognosis is not well characterized. Our study aims to evaluate the clinical characteristics and outcomes of E. coli bacteremia patients specifically in relation to biofilm forming isolates. Methods 91 E. coli bacteremia clinical isolates were consecutively collected from patients between 2013 to 2015. Virulence factor phenotypes were determined by in vitro biofilm formation, motility, and milk hydrolysis. Clinical patient data associated with the isolates were abstracted from the electronic medical records...

Research paper thumbnail of 352Clinical and molecular characteristics of NDM-1 harboring Multi-Drug Resistant Gram Negative Bacteria at Carilion Medical Center

Open Forum Infectious Diseases

Research paper thumbnail of Catheter-Based Medical Device Biofilm Ablation Using Histotripsy: A Parameter Study

Ultrasound in Medicine & Biology

Research paper thumbnail of Bacterial infection and endotoxin in female reproductive tract in rats: correlation with the developmental status of preimplantation embryos

Indian journal of experimental biology, 1998

During mammalian preimplantation development, a substantial numbers of embryos are believed to be... more During mammalian preimplantation development, a substantial numbers of embryos are believed to be lost for reasons that are unclear. Using female rats, we investigated whether the developmental status of embryos is influenced by bacterial infection and endotoxin in the reproductive tract. From the vagina of cycling rats (n = 11), 21 bacterial isolates were identified; they were Streptococcus faecalis (S. faecalis; 38%), Escherichia coli (E. coli; 19%), Acinetobactor calcoaceticus (A. calcoaceticus; 14%), and coagulase negative staphylococcus (14%), Micrococcus sp. (5%), Bacillus subtilis (B. subtilis; 5%) and Proteus vulgaris (P. vulgaris; 5%). From the vagina of day 4 pregnant rats (n = 12), 26 isolates were identified; they were S. faecalis (23%), A. calcoaceticus (23%), E. coli (15%), Micrococcus sp. (15%), B. subtilis (8%), P. vulgaris (4%), Staphylococcus aureus (4%), beta-hemolytic streptococcus (4%) and Pseudomonas aeruginosa (4%). Gram negative bacteria found in the vagina o...

Research paper thumbnail of A Novel Oxidized Low-Density Lipoprotein-Binding Protein, Asp-Hemolysin, Recognizes Lysophosphatidylcholine

Biological & Pharmaceutical Bulletin, 2002

Research paper thumbnail of Purification and characterization of esterases D-1 and D-2 from human erythrocytes

Archives of Biochemistry and Biophysics, 1988

Esterase D-1 (carboxylesterase; carboxylic-ester hydrolase, EC 3.1.1.1) was purified to homogenei... more Esterase D-1 (carboxylesterase; carboxylic-ester hydrolase, EC 3.1.1.1) was purified to homogeneity and esterase D-2 was highly purified from human erythrocytes. A new procedure, which included fractionation with ammonium sulfate, hydrophobic chromatography on a Toyopearl HW-65 column, and chromatographies on CM-cellulose and hydroxylapatite columns, was developed. Esterases D-1 and D-2 were purified about 9000- and 5600-fold over the precipitates with 65% saturated ammonium sulfate in 14 and 35% yields, respectively. The minimum molecular weights of esterases D-1 and D-2 were estimated to be 35,000 based on the mobilities on sodium dodecyl sulfate-polyacrylamide gel electrophoresis with or without 2-mercaptoethanol. The molecular weights of both enzymes were calculated to be 76,000 by gel filtration. These findings indicated that these two enzymes consisted of dimer without an intermolecular disulfide bond(s). Amino acid analysis of esterase D-1 showed that the total residues of aspartic acid plus asparagine, glutamic acid plus glutamine, glycine, and leucine represent about 40% of the total amino acid residues. Esterases D-1 and D-2 have almost identical biochemical characteristics, including Km values, sensitivities to sulfhydryl reagents, and molecular weights. Esterase D-2 cross-reacted with a rabbit antibody raised against the purified esterase D-1.

Research paper thumbnail of Rapid diagnosis of typhoid fever by a new ELISA test with porin antigens

Indian Journal of Medical Microbiology, 1993

... 1993 : 11(3), 191-196 RAPID DIAGNOSIS OF TYPHOID FEVER BY A NEW ELISA TEST WITH PORIN ANTIGEN... more ... 1993 : 11(3), 191-196 RAPID DIAGNOSIS OF TYPHOID FEVER BY A NEW ELISA TEST WITH PORIN ANTIGENS Jayasimha Rao*, M. Raghunath** and Nandan Singh ... MKJ Sarma & KM Nair of NIN for the useful discussions, and valuable help in the development of this method. ...

Research paper thumbnail of Novel Phosphorylcholine‐Containing Protein ofPseudomonas aeruginosaChronic Infection Isolates Interacts with Airway Epithelial Cells

The Journal of Infectious Diseases, 2008

Pseudomonas aeruginosa undergoes phase variation in the expression of the phosphorylcholine (ChoP... more Pseudomonas aeruginosa undergoes phase variation in the expression of the phosphorylcholine (ChoP) epitope, a structure crucial for the virulence of several respiratory pathogens. In this study, ChoP expression analysis comparing organisms from acute and chronic infections revealed that expression of ChoP at 37°C was higher among strains from chronic infections. Coimmunoprecipitation experiments and mass spectrometry analysis demonstrated that ChoP was on the protein elongation factor Tu. The presence of ChoP at the surface was confirmed by immunofluorescence and flow cytometry analysis of intact bacteria. Pretreatment of bronchial epithelial cells or mice with a platelet-activating factor receptor (PAFR) antagonist reduced adhesion and invasion of the ChoP-positive P. aeruginosa isolates. Results of this study suggest that ChoP expression may represent a novel phenotype expressed by the chronic infection isolates that could mediate P. aeruginosa colonization of the epithelial airway by means of the interaction with the PAFR.

Research paper thumbnail of RahU: An inducible and functionally pleiotropic protein in Pseudomonas aeruginosa modulates innate immunity and inflammation in host cells

Cellular Immunology, 2011

The aim of this study was to define the functional role of a recently identified RahU protein fro... more The aim of this study was to define the functional role of a recently identified RahU protein from Pseudomonas aeruginosa in macrophages and its role in bacterial defense. Recombinant (r)-RahU had no significant effect on cell apoptosis or cell viability in human monocytic THP-1 cells. Gene expression array of murine macrophage cells (RAW 264.7) stimulated with LPS showed modulation of common transcripts (by r-RahU and predisone) involved in inflammation. Functional cellular analysis showed RAW cells incubated with r-RahU at 1.0-10 μg/ml (0.06-0.6 μM) inhibited accumulation of nitric oxide (NO) in the presence of LPS by 10-50%. The IC(50) of r-RahU (0.6 μM) was distinct from the known inhibitors of NO production: prednisone (50 μM) and L-NMMA (100 μM). r-RahU also significantly inhibited chemotactic activity of THP-1 cells toward CCL2 or chemotactic supernatants from apoptotic T-cells. These reports show previously unknown pleiotropic properties of RahU in modulating both microbial physiology and host innate immunity.

Research paper thumbnail of Comparisons of Two Proteomic Analyses of Non-Mucoid and Mucoid Pseudomonas aeruginosa Clinical Isolates from a Cystic Fibrosis Patient

Frontiers in Microbiology, 2011

Research paper thumbnail of Prospects of riboflavin carrier protein (RCP) as an antifertility vaccine in male and female mammals

Human Reproduction Update, 1997

Introduction 325 RCP in the chicken oocyte 326 Evolutionary conservation 327 Structural modificat... more Introduction 325 RCP in the chicken oocyte 326 Evolutionary conservation 327 Structural modification of RCP for eliciting bioeffective antibodies 327 Effects of active immunization with SDS-RCM-RCP on reproductive performance of female Bonnet monkeys 328 Potential stage of immunointerference with pregnancy 328 Evaluation of C-terminal peptide of RCP as an immunogen 329 Evaluation of an N-terminal peptide (NTP) of RCP as the immunogen 330 RCP in male reproduction 330 Conclusions and future prospects 332 Acknowledgements 333 References 333 Riboflavin carrier protein (RCP) is obligatorily involved in yolk deposition of the vitamin, riboflavin, in the developing oocyte of the hen. The production of this protein is inducible by oestrogen. It is evolutionarily conserved in terms of its physicochemical, immunological and functional characteristics. It is the prime mediator of vitamin supply to the developing fetus in mammals, including primates. Passive immunoneutralization of the protein terminates pregnancy in rats. Active immunization of rats and bonnet monkeys with avian RCP prevents pregnancy without causing any adverse physiological effects of the mother in terms of her vitamin status, reproductive cycles or reproductive-endocrine profile. Denatured, linearized RCP is more effective in eliciting neutralizing antibodies capable of interfering with embryonic viability either before or during peri-implantation stages. Two defined stretches of sequential epitopes, one located at the N-terminus and the other at the C-terminus of the protein have been identified. Active immunization with either of these epitopes conjugated with diphtheria toxoid curtails pregnancy in rats and monkeys. Immunohistochemical localization of RCP on ovulated oocytes and early embryos shows that the antibodies cause degeneration only of early embryos. RCP is produced intra-testicularly and becomes localized on acrosomal surface of mammalian spermatozoa. Active immunization of male rats and monkeys with denatured RCP markedly reduces fertility by impairing the fertilizing potential of spermatozoa. These findings suggest that RCP, or its defined fragments, could be a novel, first generation vaccine for regulating fertility in both the sexes.

Research paper thumbnail of Host derived inflammatory phospholipids regulate rahU (PA0122) Gene, Protein, and Biofilm Formation in Pseudomonas aeruginosa

Cellular Immunology, 2011

Research paper thumbnail of Outer Dense Fiber Proteins Are Dominant Postobstruction Autoantigens in Adult Lewis Rats 1

Obstruction of the male reproductive tract commonly results in generation of antisperm autoantibo... more Obstruction of the male reproductive tract commonly results in generation of antisperm autoantibodies. However, only a few of the sperm autoantigens recognized by these antibodies have been characterized. To identify postobstruction rat sperm autoantigens, sperm proteins were separated by two-dimensional (2-D) gel electrophoresis. Spots corresponding to proteins that were stained by at least 50 % of postvasectomy rat sera on 2-D Western blots were removed from polyacrylamide gels and microsequenced by tandem mass spectrometry. From a total of 21 spots, 12 contained peptides that matched solely to either of two outer dense fiber proteins, odf1 or odf2. Six additional spots contained peptides comprising odf1 or odf2 and were accompanied by peptides representing other proteins. Only three spots lacked outer dense fiber peptides but did contain sequences of other known proteins. The results indicate that the outer dense fiber proteins odf1 and odf2 are dominant postobstruction autoantig...

Research paper thumbnail of Counter immuno electrophoresis of antibodies to porins (S.typhi) in serodiagnosis of typhoid fever

Indian Journal of Medical Microbiology, 1996

ABSTRACT The counter immunoelectrophoresis (CIEP) was applied to screen blood samples collected a... more ABSTRACT The counter immunoelectrophoresis (CIEP) was applied to screen blood samples collected at a single time point during second week (10-15 days) of fever from : 86 patients with typhoid fever (widal positive), five patients who were positive by blood culture for S.typhi, 27 other pyrexial cases (widal negative) and 29 healthy controls (widal negative). The percentage of widal and blood culture positive samples that were 63.7 percent positive for antiporin antibodies by the present CIEP method was found to be significantly higher (p<0.001) than that in healthy controls and other widal negative pyrexial cases. Based on these observations the characteristics of the CIEP method were computed and were found to show: sensitivity 63.7 percent, specificity 96.4 percent, accuracy 76.2 percent, positive predictive value 96.6 percent, and negative predictive value 62.0 percent respectively. None of the samples were positive for porins in serum by this method. The results suggest the usefulness of CIEP for antibodies to porins, as a simple rapid method for serodiagnosis of typhoid fever. However, the method does not seem to be useful for detection of porin antigen in early diagnosis of typhoid fever.

Research paper thumbnail of Complete Genome Sequence of Pseudomonas aeruginosa CMC-097, Isolated from a Ventilator-Associated Pneumonia Patient, Containing a Novel Carbapenem Resistance Class 1 Integron

Microbiology Resource Announcements, 2021

We report the complete genome of a clinical strain of Pseudomonas aeruginosa, CMC-097, which was ... more We report the complete genome of a clinical strain of Pseudomonas aeruginosa, CMC-097, which was isolated from a ventilator-associated pneumonia patient with a chronic infection. Illumina sequence reads were assembled using Geneious to yield a 7,044,064-bp circular chromosome containing a carbapenem resistance integron, In2020. ABSTRACT We report the complete genome of a clinical strain of Pseudomonas aeruginosa CMC-097, which was isolated from a ventilator-associated pneumonia patient with a chronic infection. Illumina sequence reads were assembled using Geneious to yield a 7,044,064-bp circular chromosome containing a carbapenem resistance integron, In2020.

Research paper thumbnail of Complete Genome Sequence of Pseudomonas aeruginosa CMC-115, a Clinical Strain from an Acute Ventilator-Associated Pneumonia Patient

Microbiology Resource Announcements, 2020

We report the complete genome of clinical strain Pseudomonas aeruginosa CMC-115, which was isolat... more We report the complete genome of clinical strain Pseudomonas aeruginosa CMC-115, which was isolated from an acute ventilator-associated pneumonia patient. Illumina sequencing reads were assembled using Geneious to yield a 6,375,262-bp circular chromosome that exhibited an unusual ferrichrome receptor in the pyoverdine synthesis locus and the absence of type 3 secretion system genes. ABSTRACT We report the complete genome of clinical strain Pseudomonas aeruginosa CMC-115, which was isolated from an acute ventilator-associated pneumonia patient. Illumina sequencing reads were assembled using Geneious to yield a 6,375,262-bp circular chromosome that exhibited an unusual ferrichrome receptor in the pyoverdine synthesis locus and the absence of type 3 secretion system genes.

Research paper thumbnail of Multidrug-resistant acinetobacter baumannii – plasmid-borne carbapenem and aminoglycoside co-resistance causing outbreak in Southwest Virginia

Research paper thumbnail of 1009. Biofilm Formation in Acinetobacter Baumannii Clinical Isolates

Open Forum Infectious Diseases

Background Multidrug resistant Acinetobacter baumannii (MDR-Ab) is a Gram-negative bacterium know... more Background Multidrug resistant Acinetobacter baumannii (MDR-Ab) is a Gram-negative bacterium known for causing severe nosocomial infections, attributed in part to its formation of biofilm. Siderophore is a virulence factor known to support biofilm formation by regulating iron availability. In this study, we screened 44 isolates of MDR-Ab from our Gram-negative repository to determine the strains that phenotypically form biofilm and produce siderophore. The results were compared to Pseudomonas aeruginosa PAO1, which produces both biofilm and siderophore. Methods Isolates were grown overnight in minimal M9 medium supplemented with casamino acids and hydroxyquinones at 37°C. Bacterial cells were normalized (to OD 600=0.01) and a standard diluted 10-3 tube was used in the study. A 96-well plate was inoculated with 100 microliters of each isolate in quadruplicates. This process was repeated in Tygon tubes with 50 microliters of each isolate in triplicates. The plate and Tygon tubes were ...

Research paper thumbnail of Focused Ultrasound Biofilm Ablation: Investigation of Histotripsy for the Treatment of Catheter-Associated Urinary Tract Infections (CAUTIs)

IEEE Transactions on Ultrasonics, Ferroelectrics, and Frequency Control

Urinary catheters often become contaminated with biofilms, resulting in catheter-associated urina... more Urinary catheters often become contaminated with biofilms, resulting in catheter-associated urinary tract infections (CAUTIs) that adversely affect patient outcomes. Histotripsy is a noninvasive focused ultrasound therapy previously developed for the noninvasive ablation of cancerous tumors and soft tissues. Histotripsy has also previously shown the ability to treat biofilms on glass slides and surgical meshes. Here, we investigate the potential of histotripsy for the treatment of CAUTIs for the first time in vitro. Clinically relevant catheter materials (Tygon, Silicone, and latex catheter mimics) and commonly used clinical catheters were tested to determine the feasibility of producing luminal histotripsy bubble clouds. A Pseudomonas aeruginosa (strain PA14) biofilm model was developed and tested to produce luminal biofilms in an in vitro Tygon catheter mimic. This model was treated with histotripsy to determine the ability to remove a luminal biofilm. Finally, the bactericidal effects of histotripsy were tested by treating PA14 suspended inside the Tygon catheter mimic. Results showed that histotripsy produced precise luminal cavitation within all tested catheter mimics and clinical catheters. Histotripsy treatment of a PA14 biofilm with histotripsy reduced luminal biofilm OD590 signal down to background levels. Further, the treatment of suspended PA14 in Manuscript

Research paper thumbnail of 2606. A Divergent Ferrichrome Receptor Associated with an Insertional Element (IS3) Identified on Novel Locus in Clinical Strain of Pseudomonas aeruginosa

Open Forum Infectious Diseases

Background Pseudomonas aeruginosa is an aerobic Gram-negative bacterium that causes life-threaten... more Background Pseudomonas aeruginosa is an aerobic Gram-negative bacterium that causes life-threatening acute and chronic infections in humans. Genotypic mutations and phenotypic variations are key features of its antimicrobial resistance and adaptation to the host environment. Pyoverdine associated genes and divergent receptors play a key role in acute Pseudomonas infections. This study seeks to address the heterogeneity of ferrichrome-iron receptor (fpvA) expression, its effect on pathogenicity and its propensity to cause acute infections clinically. Genetic and phenotypic variation of a clinical isolates of P. aeruginosa (PA097 and PA115) were identified by complete genome sequencing method. Methods An IRB-approved prospective study collected 38 P. aeruginosa clinical isolates and stored at Carilion Medical Center. Two genetically unrelated clinical strains were selected from tracheal aspirates: PA097 and PA115. These isolates were characterized by pyoverdine (pvd) quantification in...

Research paper thumbnail of 2603. Biofilm Formation as a Predictive Marker of Prognosis for Escherichia coli Sepsis

Open Forum Infectious Diseases

Background Escherichia coli is the Gram-negative organism most commonly associated with bloodstre... more Background Escherichia coli is the Gram-negative organism most commonly associated with bloodstream infections and death due to sepsis. Timely administration of appropriate antibiotic(s) plays a significant role in improving patient outcomes. E. coli expresses virulence factors (VFs) such as biofilm formation and motility phenotypes which play a role in bacterial attachment and dissemination by enabling immune system evasion and host migration. The role of these VFs in bacteremia prognosis is not well characterized. Our study aims to evaluate the clinical characteristics and outcomes of E. coli bacteremia patients specifically in relation to biofilm forming isolates. Methods 91 E. coli bacteremia clinical isolates were consecutively collected from patients between 2013 to 2015. Virulence factor phenotypes were determined by in vitro biofilm formation, motility, and milk hydrolysis. Clinical patient data associated with the isolates were abstracted from the electronic medical records...

Research paper thumbnail of 352Clinical and molecular characteristics of NDM-1 harboring Multi-Drug Resistant Gram Negative Bacteria at Carilion Medical Center

Open Forum Infectious Diseases