Edward Rossomando - Academia.edu (original) (raw)

Papers by Edward Rossomando

Dental Hypotheses, 2021

After W.D. Miller proved a causal relationship between microbes and dental caries and periodontit... more After W.D. Miller proved a causal relationship between microbes and dental caries and periodontitis, the repair and replacement of damaged or lost teeth resulting from microbial activity dominated 20th century dental practice. In this study, I predict that in the 21st century dental practice will shift to the treatment of those dental diseases not caused by microbes. As dentists already treat some nonmicrobial diseases, I will focus on craniofacial malformations, the group of nonmicrobial diseases usually called birth defects. Some examples include dental dysplasias, cleft lip and palate, and malocclusion. In this study, I introduce the word “dysmorphogenesis” (to replace the term birth defect) as it more appropriately ascribes this subset of nonmicrobial diseases results to mistakes during the formation of craniofacial structures. As dysmorphogenic diseases occur during gestation, their diagnosis and especially their treatment require intervention during embryogenesis. Fortunately,...

Dental Hypotheses, 2021

This paper examines the delivery of oral health care and compares the delivery in 2020 with that ... more This paper examines the delivery of oral health care and compares the delivery in 2020 with that before 1900. While equipment and materials have changed, the design of the office, and the use of personnel within the office has remained remarkably similar. The doctor is responsible for diagnosis and treatment with some support services provided by hygienists and assistants. This piece work delivery model contrasts with an assembly line model. The assembly line model is explored here in a hypothetical visit to a dental office in 2025. The paper describes the layout of practice as well as the role of the doctor and auxiliary personnel in the office.

Journal of Endodontics, 1991

Root canal samples, taken from periapical tissue exudates during routine root canal treatment pro... more Root canal samples, taken from periapical tissue exudates during routine root canal treatment procedures, were processed for identification of tumor necrosis factor using a mouse anti-human monoclonal antibody and enzyme-linked immunosorbent assay. Detectable levels of tumor necrosis factor were identified in periapical tissue exudates in chronic apical periodontitis.

Compendium of continuing education in dentistry (Jamesburg, N.J. : 1995), 2009

Compendium of continuing education in dentistry (Jamesburg, N.J. : 1995), 2005

Journal of Chromatography B: Biomedical Sciences and Applications, 1992

A batch separation procedure has been developed for retrieval of tumor necrosis factor (TNF) alph... more A batch separation procedure has been developed for retrieval of tumor necrosis factor (TNF) alpha from the microliter volumes of fluid isolated from the human temporomandibular joint (TMJ). Paramagnetic beads coated with monoclonal antibodies for TNF were used. The beads, and bound TNF, were recovered from solution with the aid of a magnetic field. The amount of bead-bound TNF was quantified using an immuno-based assay developed in this laboratory called the cluster assay. The cluster assay was specific for TNF and linear up to 10 ng. Using these methods we found that TMJ fluid contained 0.2-4.2 ng per 100 microliters of fluid with a mean value of 1.9 ng and a standard deviation of 1.1 ng. This study demonstrates the utility of batch immunomagnetic separation for the concentration and purification of proteins, and the cluster assay for quantification of proteins from microliter volumes of body fluids.

Compendium of continuing education in dentistry (Jamesburg, N.J. : 1995), 2004

New products don’t simply appear in the dental office. New products begin life as an idea, which ... more New products don’t simply appear in the dental office. New products begin life as an idea, which first must be captured so that with creativity, energy, time, and money it will be patented and become an “invention.” However, this is only the beginning. To become a product, the invention must traverse a path that is not clearly marked and where obstacles can stall its progress at every step along the way. The slow rate of introduction of new products to the dental profession suggests that the existing process for new product development needs improvement. The research to improve the process must begin at the start of the process—the idea stage. Who has the ideas, how to improve their capture, and how to encourage their conversion to inventions need to be examined. Next, research on how to transform the invention to a product is necessary. Generally, licensing, obtaining venture capital, prototype development, and obtaining regulatory approval are involved. Finally, it is crucial for the next generation of dental academics and practioners to have knowledge, experience, and skills on how to generate the new ideas and incorporate them as new products into their practice. It follows that dental school curricula should include courses in new product development and encourage dental students to participate in the research into these areas. I predict that dental school graduates who receive education and research experience in new product development will be more likely to contribute ideas for new products. They also will be more likely to accept new products into their practices. As a result, the rate of new product development will accelerate.

Nucleic Acids Research, 1997

A protocol for increasing transcription from plasmid expression vectors is presented. A vector co... more A protocol for increasing transcription from plasmid expression vectors is presented. A vector containing chloramphenicol acetyltransferase (CAT) gene was digested leaving the transcription cassette intact. Heat inactivation of restriction enzymes followed by ligation of the digestion products yielded concatemers which migrated as a single band in agarose gel electrophoresis. Mouse fibroblasts transfected with the concatemers gave a CAT activity that was 14-fold greater than that of cells transfected with a similar mass (equimolar gene number) of the native plasmid. The effect was independent of promoter type, restriction enzyme, number of restriction sites and with a noted exception, cell line.

Molecular and Cellular Biochemistry, 1986

AMP deaminase, the activity that catalyzes the deamination of AMP to form IMP and NH 3 has been m... more AMP deaminase, the activity that catalyzes the deamination of AMP to form IMP and NH 3 has been measured in Dictyostelium discoideum. A new procedure to assay the activity of this enzyme was developed using formycin 5'-monophosphate, a fluorescent analog of AMP as the substrate, and ionpaired reverse phase HPLC to separate the reactants and products. Quantitation of the formycin containing compounds was accomplished at 290 nm. At this wavelength adenosine containing compounds were not detected and activity could be monitored in the presence of its activator ATE The AMP deaminase activity in vegetative cells was 7.4 nmols/min/mg proteins while the activity in cells measured at 2 and 6 hrs after starvationinduced growth-arrest was 376 nmols/min/mg protein.., a 51-fold increase. When vegetative cells were treated with hadacidin, a drug that restricts de novo AMP synthesis and pinocytosis, the activity of the AMP deaminase was 511 nmols/min/mg protein.., a 70-fold increase compared to that in untreated vegetative cells. Smaller increases were noted following the inhibition of growth with the drugs cerulenin and vinblastine, as well as after the inhibition of de novo GMP synthesis with the drug mycophenolic acid or the partial inhibition of de novo AMP synthesis with analogs of hadacidin, N-hydroxyglycine and Nformylglycine. In addition, when the activity of two other enzymes involved in purine metabolism, namely adenosine kinase and hypoxanthine-guanine phosphoribosyl transferase, was measured in vegetative cells, and the activity of both compared to that measured in starvation and hadacidin induced growth-arrested cells, showed no significant changes. These data suggest that the changes in the activity of the AMP deaminase are in response to nutrient deprivation and further, that as a consequence of the increase in AMP deaminase activity, ammonia will be produced and an increase in pH should follow. The production of ammonia and its effect on development implicates the AMP deaminase in the early differentiation of this organism.

The Journal of Organic Chemistry, 1988

... Wei-Che Sun,+ Pamela M. Guy,' Jessica H. Jahngen,* Edward F. Rossomando,t and Ed... more ... Wei-Che Sun,+ Pamela M. Guy,' Jessica H. Jahngen,* Edward F. Rossomando,t and Edwin G. E. Jahngen*it Department of Chemistry, University of Lowell, Lowell, Massachusetts 01854, and Department of Oral Biology ... (8) Moon, S.; Duchin, L.; Cooney, JV Tetrahedron Lett. ...

FEMS Immunology & Medical Microbiology, 2007

This cross-sectional study evaluated the association between radiographic evidence of alveolar bo... more This cross-sectional study evaluated the association between radiographic evidence of alveolar bone loss and the concentration of host-derived bone resorptive factors (interleukin-1 beta, tumor necrosis factor-alpha, interleukin-6, prostaglandin-E2), and markers of bone turnover [pyridinoline cross-linked carboxyterminal telopeptide of type I collagen (ICTP), osteocalcin, osteonectin] in stimulated human whole saliva collected from 110 untreated dental patients. Alveolar bone loss scores for each patient were derived from radiographic examination. Variables positively associated with increased bone loss score were: age, current smoking, use of bisphosphonate drugs, and salivary interleukin-1beta levels above the median. Salivary osteonectin levels above the median were associated with a decreased bone loss score. Additional in vitro studies were carried out to determine the fate of interleukin-1beta, interleukin-6 and tumor necrosis factor-alpha added to whole and parotid saliva. All cytokines added to saliva were detected in significantly lower concentrations than when added to buffer alone. Protease inhibitors added to saliva did not prevent the reduction in detection of biomarkers. Variation in time of incubation, repeated cycles of freezing and thawing, or exposure to dimethylsulfoxide did not appreciably affect the measurement of cytokines in saliva. These results suggest that detection of biomarkers by conventional immunoassays may underestimate the actual quantity of molecules in saliva.

Digestive Diseases and Sciences, 1989

This study was designed to characterize neutrophil chemotactic factors released by gastric tissue... more This study was designed to characterize neutrophil chemotactic factors released by gastric tissue. Full-thickness rabbit stomach (organ culture) was prepared and incubated in Ringer's solution at 37~ Culture supernatants were collected at 1, 2, 3, and 4 hr and assayed for neutrophil chemotactic activity in modified Boyden chambers. High levels of chemotactic activity were seen at 3 hr of incubation. Antral and fundic tissue were equally capable of producing neutrophil chemotactic activity. In addition, high levels of activity were seen from both the serosal and mucosal surfaces. Initial biochemical characterization of these gastric-derived factors revealed that: (1) a majority of the activity (80-90%) exhibited molecular weight values of greater than 300 kDa, (2) the chemotactic activity Was heat stable but was partially reduced by treatment with a protease, subtilisin (37% inhibition), and (3) 70-80% of the activity in the supernatants was extracted into organic solvent (ethyl acetate). These factors may prove to be important in recruitment of neutrophils to areas of gastric injury.

Cytokine, 1996

Interaction of bacterial products and antigens from several putative periodontal pathogens with i... more Interaction of bacterial products and antigens from several putative periodontal pathogens with inflammatory cells has been reported to result in the release of cytokines such as interleukin 1, 1-3 a key mediator of various immunological and inflammatory phenomena. 4 In humans, IL-1 has been shown to exist in two distinct forms, IL-1α and IL-1β, which were encoded by separate genes. IL-1β is found to be somewhat more active than IL-1α. 5-7 A wide range of both in vitro and in vivo activity of IL-1 has been reported, including induction of PGE 2 and collagenase synthesis by cultured fibroblasts, 8-12 and increased production of IL-2 by T-lymphocyte and interferon. 13 Moreover, IL-1 is one of the factors known to stimulate bone resorption and secretion of proteinases and may be involved in the attachment loss and bone resorption which are characteristic features of periodontitis. 14-20 All of these IL-1β-dependent mechanisms may contribute to the inflammation and destruction of the bone and soft connective tissue in periodontal disease. 12,21-23 Recent clinical studies have illustrated that the amount of IL-1β is much higher in the periodontitis pockets or in the underlying inflamed gingival tissue than in healthy sites, and IL-1β is markedly reduced following treatment. 22,24-26 The presence of cytokines may be detected earlier than the appearance of acute symptoms of periodontal disease. 27,28 Measurement of IL-1β in GCF or tissue of periodontitis patients has been suggested as an important aid to monitor the severity of disease.

Bioconjugate Chemistry, 1996

A new method for coupling proteins to plasmid expression vectors is presented. Biotin was covalen... more A new method for coupling proteins to plasmid expression vectors is presented. Biotin was covalently attached to a plasmid expression vector containing a chloramphenicol acetyltransferase (CAT) gene. The specific label was one biotin per 100 bp. An electrophoretic mobility shift assay showed that the plasmid was capable of binding multiple streptavidin molecules. When transfected into mouse fibroblasts, the biotinylated plasmid retained 40% of the native plasmid's biological activity, as determined by CAT assay, and was not affected by the binding of streptavidin. The method allows for attachment of any protein to plasmid DNA expression vector while retaining biological function. Hybrid plasmids in which the transcription cassettes were kept free of biotin label were constructed by digesting biotinylated and unbiotinylated plasmids at sites outside the transcription cassette and re-ligating the digestion products. Electron microscopy studies show that the ligation products formed large tangled assemblages of plasmid DNA. When equimolar (with respect to gene number) amounts of these large hybrid biotinylated plasmids were transfected into mouse fibroblasts by means of calcium phosphate precipitation, an increase in CAT expression 25-fold greater than that of original biotinylated plasmid was observed. Slot-blot analysis of total DNA extracted from transfected cells shows that this enhanced activity was not due to increased transfection efficiency. Receptor-mediated delivery could not be shown when a complex comprising biotinylated asialoglycoprotien/streptavidin/biotinylated CAT expression vector was placed in media containing Hep G2 cells.

Biochemical and Biophysical Research Communications, 1970

Abstract Myosin monomer can be separated from aggregated myosin and heterogeneous low molecular w... more Abstract Myosin monomer can be separated from aggregated myosin and heterogeneous low molecular weight contaminants by chromatography on 4% or 2% agarose. The monomer was characterized by its ATPase activity, which was constant across the peak, and its molecular weight (about 465,000). Chromatographic studies of aged samples show that aggregation is accompanied by the release of low molecular weight material. Molecular sieve chromatography provides a new approach to the purification of myosin which should be useful in studies on its molecular structure.

Biochemical and Biophysical Research Communications, 1972

Abstract In this report we describe the identification and preliminary characterization of an ade... more Abstract In this report we describe the identification and preliminary characterization of an adenyl cyclase in cell free extracts of D. discoideum . The specific activity of this enzyme was found to remain constant throughout growth and fruiting body construction, and the enzyme does not appear to turn over at an appreciable rate at any time. Its peculiar reaction kinetics as observed in crude extracts offer one possible explanation of the pulsating feature of the chemotactic attraction (1). In addition, a c-AMP stimulated disappearance of ATP has been noted. At least part of the latter can be ascribed to the action of a phosphokinase.

Dental Hypotheses, 2021

After W.D. Miller proved a causal relationship between microbes and dental caries and periodontit... more After W.D. Miller proved a causal relationship between microbes and dental caries and periodontitis, the repair and replacement of damaged or lost teeth resulting from microbial activity dominated 20th century dental practice. In this study, I predict that in the 21st century dental practice will shift to the treatment of those dental diseases not caused by microbes. As dentists already treat some nonmicrobial diseases, I will focus on craniofacial malformations, the group of nonmicrobial diseases usually called birth defects. Some examples include dental dysplasias, cleft lip and palate, and malocclusion. In this study, I introduce the word “dysmorphogenesis” (to replace the term birth defect) as it more appropriately ascribes this subset of nonmicrobial diseases results to mistakes during the formation of craniofacial structures. As dysmorphogenic diseases occur during gestation, their diagnosis and especially their treatment require intervention during embryogenesis. Fortunately,...

Dental Hypotheses, 2021

This paper examines the delivery of oral health care and compares the delivery in 2020 with that ... more This paper examines the delivery of oral health care and compares the delivery in 2020 with that before 1900. While equipment and materials have changed, the design of the office, and the use of personnel within the office has remained remarkably similar. The doctor is responsible for diagnosis and treatment with some support services provided by hygienists and assistants. This piece work delivery model contrasts with an assembly line model. The assembly line model is explored here in a hypothetical visit to a dental office in 2025. The paper describes the layout of practice as well as the role of the doctor and auxiliary personnel in the office.

Journal of Endodontics, 1991

Root canal samples, taken from periapical tissue exudates during routine root canal treatment pro... more Root canal samples, taken from periapical tissue exudates during routine root canal treatment procedures, were processed for identification of tumor necrosis factor using a mouse anti-human monoclonal antibody and enzyme-linked immunosorbent assay. Detectable levels of tumor necrosis factor were identified in periapical tissue exudates in chronic apical periodontitis.

Compendium of continuing education in dentistry (Jamesburg, N.J. : 1995), 2009

Compendium of continuing education in dentistry (Jamesburg, N.J. : 1995), 2005

Journal of Chromatography B: Biomedical Sciences and Applications, 1992

A batch separation procedure has been developed for retrieval of tumor necrosis factor (TNF) alph... more A batch separation procedure has been developed for retrieval of tumor necrosis factor (TNF) alpha from the microliter volumes of fluid isolated from the human temporomandibular joint (TMJ). Paramagnetic beads coated with monoclonal antibodies for TNF were used. The beads, and bound TNF, were recovered from solution with the aid of a magnetic field. The amount of bead-bound TNF was quantified using an immuno-based assay developed in this laboratory called the cluster assay. The cluster assay was specific for TNF and linear up to 10 ng. Using these methods we found that TMJ fluid contained 0.2-4.2 ng per 100 microliters of fluid with a mean value of 1.9 ng and a standard deviation of 1.1 ng. This study demonstrates the utility of batch immunomagnetic separation for the concentration and purification of proteins, and the cluster assay for quantification of proteins from microliter volumes of body fluids.

Compendium of continuing education in dentistry (Jamesburg, N.J. : 1995), 2004

New products don’t simply appear in the dental office. New products begin life as an idea, which ... more New products don’t simply appear in the dental office. New products begin life as an idea, which first must be captured so that with creativity, energy, time, and money it will be patented and become an “invention.” However, this is only the beginning. To become a product, the invention must traverse a path that is not clearly marked and where obstacles can stall its progress at every step along the way. The slow rate of introduction of new products to the dental profession suggests that the existing process for new product development needs improvement. The research to improve the process must begin at the start of the process—the idea stage. Who has the ideas, how to improve their capture, and how to encourage their conversion to inventions need to be examined. Next, research on how to transform the invention to a product is necessary. Generally, licensing, obtaining venture capital, prototype development, and obtaining regulatory approval are involved. Finally, it is crucial for the next generation of dental academics and practioners to have knowledge, experience, and skills on how to generate the new ideas and incorporate them as new products into their practice. It follows that dental school curricula should include courses in new product development and encourage dental students to participate in the research into these areas. I predict that dental school graduates who receive education and research experience in new product development will be more likely to contribute ideas for new products. They also will be more likely to accept new products into their practices. As a result, the rate of new product development will accelerate.

Nucleic Acids Research, 1997

A protocol for increasing transcription from plasmid expression vectors is presented. A vector co... more A protocol for increasing transcription from plasmid expression vectors is presented. A vector containing chloramphenicol acetyltransferase (CAT) gene was digested leaving the transcription cassette intact. Heat inactivation of restriction enzymes followed by ligation of the digestion products yielded concatemers which migrated as a single band in agarose gel electrophoresis. Mouse fibroblasts transfected with the concatemers gave a CAT activity that was 14-fold greater than that of cells transfected with a similar mass (equimolar gene number) of the native plasmid. The effect was independent of promoter type, restriction enzyme, number of restriction sites and with a noted exception, cell line.

Molecular and Cellular Biochemistry, 1986

AMP deaminase, the activity that catalyzes the deamination of AMP to form IMP and NH 3 has been m... more AMP deaminase, the activity that catalyzes the deamination of AMP to form IMP and NH 3 has been measured in Dictyostelium discoideum. A new procedure to assay the activity of this enzyme was developed using formycin 5'-monophosphate, a fluorescent analog of AMP as the substrate, and ionpaired reverse phase HPLC to separate the reactants and products. Quantitation of the formycin containing compounds was accomplished at 290 nm. At this wavelength adenosine containing compounds were not detected and activity could be monitored in the presence of its activator ATE The AMP deaminase activity in vegetative cells was 7.4 nmols/min/mg proteins while the activity in cells measured at 2 and 6 hrs after starvationinduced growth-arrest was 376 nmols/min/mg protein.., a 51-fold increase. When vegetative cells were treated with hadacidin, a drug that restricts de novo AMP synthesis and pinocytosis, the activity of the AMP deaminase was 511 nmols/min/mg protein.., a 70-fold increase compared to that in untreated vegetative cells. Smaller increases were noted following the inhibition of growth with the drugs cerulenin and vinblastine, as well as after the inhibition of de novo GMP synthesis with the drug mycophenolic acid or the partial inhibition of de novo AMP synthesis with analogs of hadacidin, N-hydroxyglycine and Nformylglycine. In addition, when the activity of two other enzymes involved in purine metabolism, namely adenosine kinase and hypoxanthine-guanine phosphoribosyl transferase, was measured in vegetative cells, and the activity of both compared to that measured in starvation and hadacidin induced growth-arrested cells, showed no significant changes. These data suggest that the changes in the activity of the AMP deaminase are in response to nutrient deprivation and further, that as a consequence of the increase in AMP deaminase activity, ammonia will be produced and an increase in pH should follow. The production of ammonia and its effect on development implicates the AMP deaminase in the early differentiation of this organism.

The Journal of Organic Chemistry, 1988

... Wei-Che Sun,+ Pamela M. Guy,' Jessica H. Jahngen,* Edward F. Rossomando,t and Ed... more ... Wei-Che Sun,+ Pamela M. Guy,' Jessica H. Jahngen,* Edward F. Rossomando,t and Edwin G. E. Jahngen*it Department of Chemistry, University of Lowell, Lowell, Massachusetts 01854, and Department of Oral Biology ... (8) Moon, S.; Duchin, L.; Cooney, JV Tetrahedron Lett. ...

FEMS Immunology & Medical Microbiology, 2007

This cross-sectional study evaluated the association between radiographic evidence of alveolar bo... more This cross-sectional study evaluated the association between radiographic evidence of alveolar bone loss and the concentration of host-derived bone resorptive factors (interleukin-1 beta, tumor necrosis factor-alpha, interleukin-6, prostaglandin-E2), and markers of bone turnover [pyridinoline cross-linked carboxyterminal telopeptide of type I collagen (ICTP), osteocalcin, osteonectin] in stimulated human whole saliva collected from 110 untreated dental patients. Alveolar bone loss scores for each patient were derived from radiographic examination. Variables positively associated with increased bone loss score were: age, current smoking, use of bisphosphonate drugs, and salivary interleukin-1beta levels above the median. Salivary osteonectin levels above the median were associated with a decreased bone loss score. Additional in vitro studies were carried out to determine the fate of interleukin-1beta, interleukin-6 and tumor necrosis factor-alpha added to whole and parotid saliva. All cytokines added to saliva were detected in significantly lower concentrations than when added to buffer alone. Protease inhibitors added to saliva did not prevent the reduction in detection of biomarkers. Variation in time of incubation, repeated cycles of freezing and thawing, or exposure to dimethylsulfoxide did not appreciably affect the measurement of cytokines in saliva. These results suggest that detection of biomarkers by conventional immunoassays may underestimate the actual quantity of molecules in saliva.

Digestive Diseases and Sciences, 1989

This study was designed to characterize neutrophil chemotactic factors released by gastric tissue... more This study was designed to characterize neutrophil chemotactic factors released by gastric tissue. Full-thickness rabbit stomach (organ culture) was prepared and incubated in Ringer's solution at 37~ Culture supernatants were collected at 1, 2, 3, and 4 hr and assayed for neutrophil chemotactic activity in modified Boyden chambers. High levels of chemotactic activity were seen at 3 hr of incubation. Antral and fundic tissue were equally capable of producing neutrophil chemotactic activity. In addition, high levels of activity were seen from both the serosal and mucosal surfaces. Initial biochemical characterization of these gastric-derived factors revealed that: (1) a majority of the activity (80-90%) exhibited molecular weight values of greater than 300 kDa, (2) the chemotactic activity Was heat stable but was partially reduced by treatment with a protease, subtilisin (37% inhibition), and (3) 70-80% of the activity in the supernatants was extracted into organic solvent (ethyl acetate). These factors may prove to be important in recruitment of neutrophils to areas of gastric injury.

Cytokine, 1996

Interaction of bacterial products and antigens from several putative periodontal pathogens with i... more Interaction of bacterial products and antigens from several putative periodontal pathogens with inflammatory cells has been reported to result in the release of cytokines such as interleukin 1, 1-3 a key mediator of various immunological and inflammatory phenomena. 4 In humans, IL-1 has been shown to exist in two distinct forms, IL-1α and IL-1β, which were encoded by separate genes. IL-1β is found to be somewhat more active than IL-1α. 5-7 A wide range of both in vitro and in vivo activity of IL-1 has been reported, including induction of PGE 2 and collagenase synthesis by cultured fibroblasts, 8-12 and increased production of IL-2 by T-lymphocyte and interferon. 13 Moreover, IL-1 is one of the factors known to stimulate bone resorption and secretion of proteinases and may be involved in the attachment loss and bone resorption which are characteristic features of periodontitis. 14-20 All of these IL-1β-dependent mechanisms may contribute to the inflammation and destruction of the bone and soft connective tissue in periodontal disease. 12,21-23 Recent clinical studies have illustrated that the amount of IL-1β is much higher in the periodontitis pockets or in the underlying inflamed gingival tissue than in healthy sites, and IL-1β is markedly reduced following treatment. 22,24-26 The presence of cytokines may be detected earlier than the appearance of acute symptoms of periodontal disease. 27,28 Measurement of IL-1β in GCF or tissue of periodontitis patients has been suggested as an important aid to monitor the severity of disease.

Bioconjugate Chemistry, 1996

A new method for coupling proteins to plasmid expression vectors is presented. Biotin was covalen... more A new method for coupling proteins to plasmid expression vectors is presented. Biotin was covalently attached to a plasmid expression vector containing a chloramphenicol acetyltransferase (CAT) gene. The specific label was one biotin per 100 bp. An electrophoretic mobility shift assay showed that the plasmid was capable of binding multiple streptavidin molecules. When transfected into mouse fibroblasts, the biotinylated plasmid retained 40% of the native plasmid's biological activity, as determined by CAT assay, and was not affected by the binding of streptavidin. The method allows for attachment of any protein to plasmid DNA expression vector while retaining biological function. Hybrid plasmids in which the transcription cassettes were kept free of biotin label were constructed by digesting biotinylated and unbiotinylated plasmids at sites outside the transcription cassette and re-ligating the digestion products. Electron microscopy studies show that the ligation products formed large tangled assemblages of plasmid DNA. When equimolar (with respect to gene number) amounts of these large hybrid biotinylated plasmids were transfected into mouse fibroblasts by means of calcium phosphate precipitation, an increase in CAT expression 25-fold greater than that of original biotinylated plasmid was observed. Slot-blot analysis of total DNA extracted from transfected cells shows that this enhanced activity was not due to increased transfection efficiency. Receptor-mediated delivery could not be shown when a complex comprising biotinylated asialoglycoprotien/streptavidin/biotinylated CAT expression vector was placed in media containing Hep G2 cells.

Biochemical and Biophysical Research Communications, 1970

Abstract Myosin monomer can be separated from aggregated myosin and heterogeneous low molecular w... more Abstract Myosin monomer can be separated from aggregated myosin and heterogeneous low molecular weight contaminants by chromatography on 4% or 2% agarose. The monomer was characterized by its ATPase activity, which was constant across the peak, and its molecular weight (about 465,000). Chromatographic studies of aged samples show that aggregation is accompanied by the release of low molecular weight material. Molecular sieve chromatography provides a new approach to the purification of myosin which should be useful in studies on its molecular structure.

Biochemical and Biophysical Research Communications, 1972

Abstract In this report we describe the identification and preliminary characterization of an ade... more Abstract In this report we describe the identification and preliminary characterization of an adenyl cyclase in cell free extracts of D. discoideum . The specific activity of this enzyme was found to remain constant throughout growth and fruiting body construction, and the enzyme does not appear to turn over at an appreciable rate at any time. Its peculiar reaction kinetics as observed in crude extracts offer one possible explanation of the pulsating feature of the chemotactic attraction (1). In addition, a c-AMP stimulated disappearance of ATP has been noted. At least part of the latter can be ascribed to the action of a phosphokinase.