Background: All-trans-retinoyl beta-D-glucuronide, a water-soluble glucuronic acid conjugate of a... more Background: All-trans-retinoyl beta-D-glucuronide, a water-soluble glucuronic acid conjugate of all-trans-retinoic acid, has demonstrated high biological activity and low toxicity in most in vitro and in vivo models. Since the reparative effects of retinoids on epithelium are well-known, our aim was to study the effect(s) of intravenously-administered all-trans-retinoyl beta-D-glucuronide in lambs with chronic bacterial bronchopneumonia. Material/methods: Two groups of lambs were inoculated intrabronchially with either pyrogen-free saline or Mannheimia haemolytica. Thirty-three days later, lambs were injected four times at five-day intervals with 2 mL of 116 mM all-trans-retinoyl beta-D-glucuronide (6.0-9.3 mmol/kg or 2.86-4.42 mg/kg animal body weight) in dimethyl sulfoxide, or dimethyl sulfoxide alone. Animal behavior and signs of clinical illness were logged daily. Lung and liver samples were assessed for histopathology, while serum and liver samples were extracted for retinoids and analyzed by reversed-phase gradient high-performance liquid chromatography. Results: Repeated injections of highly concentrated all-trans-retinoyl beta-D-glucuronide did not cause changes in appetite, activity or other behaviors nor did it cause histologic lesions in liver and lung. However, it had no effect on resolution of lung lesions resultant of chronic Mannheimia haemolytica bronchopneumonia. Conclusions: Repeated intravenous administration of high amounts of all-trans-retinoyl beta-D-glucuronide to lambs did not elicit signs of gross or microscopic toxicity. However, administering all-trans-retinoyl beta-D-glucuronide too late in the progression of bacterial pneumonia is thought to be the main reason for its lack of effect in this model. A retinoid lactone derivative was detected in sheep serum and liver several days after treatment.
Medical science monitor : international medical journal of experimental and clinical research, 2002
All-trans-retinoyl beta-D-glucuronide, a water-soluble glucuronic acid conjugate of all-trans-ret... more All-trans-retinoyl beta-D-glucuronide, a water-soluble glucuronic acid conjugate of all-trans-retinoic acid, has demonstrated high biological activity and low toxicity in most in vitro and in vivo models. Since the reparative effects of retinoids on epithelium are well-known, our aim was to study the effect(s) of intravenously-administered all-trans-retinoyl beta-D-glucuronide in lambs with chronic bacterial bronchopneumonia. Two groups of lambs were inoculated intrabronchially with either pyrogen-free saline or Mannheimia haemolytica. Thirty-three days later, lambs were injected four times at five-day intervals with 2 mL of 116 mM all-trans-retinoyl beta-D-glucuronide (6.0-9.3 mmol/kg or 2.86-4.42 mg/kg animal body weight) in dimethyl sulfoxide, or dimethyl sulfoxide alone. Animal behavior and signs of clinical illness were logged daily. Lung and liver samples were assessed for histopathology, while serum and liver samples were extracted for retinoids and analyzed by reversed-phase...
Treatment of root canals with ultraviolet C (UVC) light-emitting diodes (LED) is a promising adju... more Treatment of root canals with ultraviolet C (UVC) light-emitting diodes (LED) is a promising adjunct to current root canal disinfection procedures that are essential for preserving the health and the healing of periapical tissues. Here, we hypothesized that UVC LED treatment would kill microorganisms associated with endodontic infections and kill microorganisms in instrumented root canal models. Three different UVC LED units were constructed. One unit emitted 265 nm from a 12 mW LED, one unit emitted 265 nm from a 22.5 mW LED, and one unit emitted 280 nm from an 8 mW LED. Power levels emitted from the three units were measured without and with fiberoptic (FO) filaments using an energy meter (PM100D, Thorlabs, Inc., Newton, NJ USA) with a standard photodiode sensor (S120VC, 200-1100 nm, 50 mW; Thorlabs, Newton, NJ USA) for 30 seconds. The raw data from the energy meter readings for these 3 units is listed in Table 1. This includes the wavelength the meter used to record the data for each unit (Wavelength read at (nm)), the distance the unit was from the sensor (Distance from sensor (mm)), the minimum power value recorded from the meter (Minimum value (µW)), the maximum power value recorded from the meter (Maximum value (µW)), the mean value recorded from the meter (Mean value (µW)), the standard deviation of the mean value recorded from the meter (Standard deviation (µW)), the number of meter readings (Number of readings (n)), and the time of recording (Time (sec)). The raw data from the antimicrobial activities of these 3 units on Candida albicans ATCC 64124, Enterococcus faecalis ATCC 29212, Streptococcus sanguinis ATCC 10556, Staphylococcus aureus ATCC 29213, and methicillin-resistant S. aureus (MRSA #7) treated on discs in laboratory assays is listed in Table 2. This includes the length of time each microorganism was treated (Group (seconds)), the UVC LED unit used to treat each microorganism (UVC LED treatment), the replication number (Replication), the microorganism treated (Microorganisms), and the numbe [...]
Although some vaccines and antibiotics have been very effective in preventing and treating respir... more Although some vaccines and antibiotics have been very effective in preventing and treating respiratory disease, they have not been fully satisfactory. Recently, components of the innate immune system have been increasingly appreciated for their role in host defense against microbial pathogens. These molecules include lysozyme and lactoferrin, but recent work in cattle, sheep, man and other species have identified new classes of peptides expressed by respiratory epithelial cells that have potent microcidal activity in nanogram quantities. These peptides, termed antimicrobial peptides (AMP), include defensins, cathelicidins and anionic peptides. Some are expressed continuously whereas expression of others is stimulated by infection/inflammation. In calves, we have found that two AMP from the defensin family, tracheal antimicrobial peptide (TAP) and lingual antimicrobial peptide (LAP), are expressed in the newborn and increased in response to Mannheimia (Pasteurella) haemolytica infection. In contrast, sheep beta defensin-1 (SBD-1) is not induced by infection and, in fact, appears decreased during viral infection with paramyxovirus-3 (PI-3). Decreased SBD-1 by Pl-3 may increase the lung's susceptibility to secondary infections or re-infections. Other innate defense molecules include proteins released with lung surfactant. Surfactant protein A and D (SAD) can opsonize and aggregate respiratory syncytial virus (RSV) and activate alveolar macrophages. Preliminary work suggests that chronic bacterial infections result in reduced SpA and SpD expression and we are currently assessing SAD expression in response to Pl-3 and RSV. A long-term goal of our work is to identify ways to up-regulate expression of AMP and/or surfactant proteins in the neonate and at times of stress in older animals in order to r educe microbial colonization. Other investigators are seeking ways to utilize AMP's as a new class of antibiotics .
American Journal of Veterinary Research, May 1, 2001
Objectives-To determine effects of selectin inhibitor TBC1269 on neutrophil infiltration, and neu... more Objectives-To determine effects of selectin inhibitor TBC1269 on neutrophil infiltration, and neutrophilassociated injury during pneumonia induced by Mannheimia haemolytica and concentration of antimicrobial anionic peptide (AAP) in bronchoalveolar lavage fluid (BALF) as well as antimicrobial activity of BALF from healthy (control) neonatal calves, neonatal calves with M haemolytica-induced pneumonia, neonatal calves with prior treatment with TBC1269, and adult cattle. Animals-Eighteen 1-to 3-day-old calves and 9 adult cattle. Procedure-Calves were inoculated with M haemolytica or pyrogen-free saline (0.14M NaCl) solution into the right cranial lung lobe, and BALF was collected 2 or 6 hours after inoculation. Thirty minutes before and 2 hours after inoculation, 4 calves received TBC1269. The BALF collected from 9 adult cattle was used for comparison of BALF AAP concentration and antimicrobial activity. Protein concentration and neutrophil differential percentage and degeneration in BALF were determined. An ELISA and killing assay were used to determine BALF AAP concentration and antimicrobial activity, respectively. Results-Total protein concentration was significantly decreased in BALF from calves receiving TBC1269. Similar concentrations of AAP were detected in BALF from all calves, which were 3-fold higher than those in BALF from adult cattle. However, BALF from neonates had little or no anti-M haemolytica activity. Conclusions and Clinical Relevance-These results suggest that TBC1269 decreases pulmonary tissue injury in neonatal calves infected with M haemolytica. Although AAP is detectable in neonatal BALF at 3 times the concentration detected in adult BALF, neonatal BALF lacks antimicrobial activity for M haemolytica.
FREE-LIVING AND HOST-ASSOCIATED PROTOZOA AS TRAINING CAMPS FOR INTRACELLULAR PATHOGENS Steve A. C... more FREE-LIVING AND HOST-ASSOCIATED PROTOZOA AS TRAINING CAMPS FOR INTRACELLULAR PATHOGENS Steve A. Carlson, Sharon K. Franklin, and Mark A. Rasmussen 9 In 1976, a group of American Legion members gathered in Philadelphia, PA, for a convention. ...
Ovine progressive pneumonia is caused by a lentivirus of known infectivity only for sheep and goa... more Ovine progressive pneumonia is caused by a lentivirus of known infectivity only for sheep and goats. Virus susceptibility of 11 other species of animals was examined. Species included cattle, chickens, deer, dogs, goats, hamsters, horses, mice, pigs, rabbits, and rats. Of these species, only goats and rabbits could be experimentally infected with the virus. The infection in rabbits was acute, and virus did not persist or induce antibody production as it does in sheep and goats. Sera obtained from several people working in close contact with the virus and from several wild species, with unknown exposure history, were tested for antibodies to viral antigens. All results were negative. Knowledge of the host range of this virus is important for scientific studies and for virus eradication programs.
Cell-mediated immune mechanisms may play a role in the pathogenesis and prevention of pneumonia i... more Cell-mediated immune mechanisms may play a role in the pathogenesis and prevention of pneumonia in cattle caused by Pasteurella haemolytica serotype A1. To determine the circumstances required to stimulate and identify cell-mediated immune responses, calves were vaccinated with a commercial P. haemolytica bacterin or a live commercial P. haemolytica vaccine, or were infected intratracheally with virulent P. haemolytica. All calves were challenge-exposed intratracheally with P. haemolytica 31 d after vaccination or prior infection. Peripheral blood mononuclear cells and mediastinal and superficial cervical lymph node cells were stimulated with antigens prepared from P. haemolytica to evaluate in vitro proliferative responses and gamma-interferon production as measures of cell-mediated immunity. Strong proliferative responses and gamma-interferon production were detected in lymph node cells from calves vaccinated with the live vaccine and from infected calves, especially in response to stimulation with an outer membrane protein preparation from P. haemolytica. Greater proliferative responses and gamma-interferon production were associated with the lymph node nearer the site of bacterin administration (superficial cervical lymph node) or the site of infection (mediastinal lymph node), whereas greater proliferative responses and gamma-interferon production were associated with the more distant lymph node (mediastinal lymph node) in calves vaccinated with the live vaccine. Neither proliferative responses nor gamma-interferon production were detected in peripheral blood mononuclear cells from calves that were vaccinated for or infected with P. haemolytica. Antileukotoxin antibody titers were determined by a serum neutralization assay, and protection against pneumonic lesions was more closely correlated with antileukotoxin antibody responses than with lymphocyte proliferation or gamma-interferon responses.
A field trial to evaluate a whole cell vaccine for the prevention of caseous lymphadenitis (CLA) ... more A field trial to evaluate a whole cell vaccine for the prevention of caseous lymphadenitis (CLA) in sheep and goats was performed in one goat herd and one sheep flock over a period of three years. In goats, there was a nonstatistically significant trend for fewer cases of CLA in the vaccinated animals compared to the controls. In sheep, from six months to 36 months postinitial vaccination, the proportion of vaccinated sheep that developed CLA was significantly less (p less than 0.05) than in the control sheep. The antibody titers to Corynebacterium pseudotuberculosis as detected by microagglutination assay were significantly different (p less than 0.0001) at all times except at the initial vaccination. Swellings occurred at the vaccination site at an incidence level of 29.6% in goats and 34.1% in sheep. The vaccine appeared to be efficacious in reducing the proportion of sheep that developed CLA when challenged naturally in a field situation.
In Alberta, caseous lymphadenitis (CLA) is one of the leading causes of lamb and mutton carcass c... more In Alberta, caseous lymphadenitis (CLA) is one of the leading causes of lamb and mutton carcass condemnation. In this study, serologic results confirmed a high (50-94%) incidence of exposure to Corynebacterium pseudotuberculosis, the causative agent of CLA, in mature, unvaccinated sheep in southern Alberta. To assess the efficacy and impact of vaccination with 2 commercial (Glanvac-6 and Case-Vac) and 1 experimental (WC+ MDP-GDP) CLA vaccines, a series of 3 field trials in 3249 ewes and lambs was conducted in affected flocks from 1992-1996. Efficacy was assessed from the serological response to vaccination, prevalence and size of injection site reactions by treatment, and the incidence of CLA abscesses. Overall, agglutinating antibody titres to C. pseudotuberculosis in lambs vaccinated with WC+MDP-GDP and Case-Vac remained significantly elevated above nonvaccinated control lambs for the 12 mo period after the initial vaccination. Lambs vaccinated with the WC/MDP-GDP maintained higher titres (P < 0.06) than those vaccinated with Case-Vac for the period from 6 to 12 mo after vaccination. Agglutinating antibody titres for lambs vaccinated with Glanvac did not differ from those of controls at any point during the 12 mo period after vaccination. The number of injection site reactions was elevated in lambs vaccinated with Glanvac as compared to those vaccinated with WC+MDP-GDP but the size of injection site reactions did not significantly differ. Sheep vaccinated with WC+ MDP-GDP also had a reduced incidence of putative CLA abscesses, although confirmation of the presence of C. pseudotuberculosis was only successful in a small number of instances.
Almost two decades after identification of the CFTR gene, we lack answers to many questions about... more Almost two decades after identification of the CFTR gene, we lack answers to many questions about the pathogenesis of cystic fibrosis (CF), and it remains a lethal disease. Mice with a disrupted CFTR gene have greatly facilitated CF studies, but they fail to develop the characteristic pancreatic, lung, intestinal, liver, and other CF manifestations. Therefore, we produced pigs with a targeted disruption of both CFTR alleles. These animals exhibited defective chloride transport. They also developed meconium ileus, exocrine pancreatic destruction, and focal biliary cirrhosis, replicating abnormalities seen in newborn patients with CF. This swine model may provide opportunities to address persistent questions about CF pathogenesis and accelerate discovery of treatments and preventions. Understanding human disease often requires animal models. Mice have been the overwhelming species of choice because methods for specifically altering their genome have
Although some vaccines and antibiotics have been very effective in preventing and treating respir... more Although some vaccines and antibiotics have been very effective in preventing and treating respiratory disease, they have not been fully satisfactory. Recently, components of the innate immune system have been increasingly appreciated for their role in host defense against microbial pathogens. These molecules include lysozyme and lactoferrin, but recent work in cattle, sheep, man and other species have identified new classes of peptides expressed by respiratory epithelial cells that have potent microcidal activity in nanogram quantities. These peptides, termed antimicrobial peptides (AMP), include defensins, cathelicidins and anionic peptides. Some are expressed continuously whereas expression of others is stimulated by infection/inflammation. In calves, we have found that two AMP from the defensin family, tracheal antimicrobial peptide (TAP) and lingual antimicrobial peptide (LAP), are expressed in the newborn and increased in response to Mannheimia (Pasteurella) haemolytica infection. In contrast, sheep beta defensin-1 (SBD-1) is not induced by infection and, in fact , appears decreased during viral infection with paramyxovirus-3 (Pl-3). Decreased SBD-1 by Pl-3 may increase the lung's susceptibility to secondary infections or re-infections. Other innate defense molecules include proteins released with lung surfactant. Surfactant protein A and D (SAD) can opsonize and aggregate respiratory syncytial virus (RSV) and activate alveolar macrophages. Preliminary work suggests that chronic bacterial infections result in reduced SpA and SpD expression and we are currently assessing SAD expression in response to PI-3 and RSV. A long-term goal of our work is to identify ways to up-regulate expression of AMP and/or surfactant proteins in the neonate and at times of stress in older animals in order to reduce microbial colonization. Other investigators are seeking ways to utilize AMP's as a new class of antibiotics .
CXCL10 (IP-10) is a small 10 kDa chemokine with antimicrobial activity. It is induced by IFN-γ, c... more CXCL10 (IP-10) is a small 10 kDa chemokine with antimicrobial activity. It is induced by IFN-γ, chemoattracts mononuclear cells, and promotes adhesion of T cells. Recently, we detected CXCL10 on the surface of the skin and in the oral cavity. In the current study, we used broth microdilution and radial diffusion assays to show that CXCL10 inhibits the growth of Escherichia coli, Staphylococcus aureus, Corynebacterium jeikeium, Corynebacterium striatum, and Candida albicans HMV4C, but not Corynebacterium bovis, Streptococcus mutans, Streptococcus mitis, Streptococcus sanguinis, Fusobacterium nucleatum, Aggregatibacter actinomycetemcomitans, Poryphromonas gingivalis, or C. albicans ATCC 64124. The reason for the selective antimicrobial activity is not yet known. However, antimicrobial activity of CXCL10 may be related to its composition and structure, as a
Ultraviolet C (UVC) light emitting diode (LED) can kill the endodontic pathogen Enterococcus faec... more Ultraviolet C (UVC) light emitting diode (LED) can kill the endodontic pathogen Enterococcus faecalis and has the potential to kill other oral microorganisms associated with endodontic infections. This same bacteriocidal device shows great promise in the stimulation of periapical healing and pain reduction resulting from inflammation in root canals. Previously, we found that 255 nm UVC LED killed E. faecalis and induced the production of cellular biomarkers in HEPM cells and gingival fibroblasts (Morio et al., 2019). Here, we extend those findings and hypothesize that UVC LED at other wavelengths and power levels kill microorganisms associated with root canal infections. Units emitting UVC LED at 265 nm (12 mW), 265 nm (22.5 mW), and 280 nm (8 mW) wavelenths were assembled and the energy levels of their emissions were measured. The energy doses in millijoules (mJ) were calculated from the power readings of the meter (µW) × time of exposure (seconds). Ex vivo models of root canals were prepared in extracted, instrumented, single canal human premolars. Five cultures of microorganisms were treated with 265 nm (12 mW), 265 nm (22.5 mW), or 280 nm (8 mW) UVC LED on discs in laboratory assays and 4 cultures of microorganisms were treated with 265 nm (22.5 mW) UVC LED in root canals of extracted, instrumented teeth. After UVC LED treatment, all microorganisms were cultivated on microbiological media. Colony forming units (CFU) of viable microorganisms treated with UVC LED were counted and compared with those of viable microorganisms not treated with UVC LED as controls. Tukey's Honestly Significant Difference was used to determine statistical significances (0.05). Units emitting UVC LED at 265 nm (12 mW), 265 nm (22.5 mW), and 280 nm (8 mW) killed Candida albicans, Staphylococcus aureus, methicillin-resistant S. aureus (MRSA), E. faecalis, and Streptococcus sanguinis after 30-90 seconds of exposure in laboratory assays (p < 0.05). Microbial killing differed among treatment times, UVC LED wavelengths, power levels of each unit, and specific microorganism. The unit emitting UVC LED at 265 nm (22.5 mW) killed C. albicans, S. aureus, MRSA, and E. faecalis in 30 s in root canals of extracted, instrumented teeth (p < 0.05). This dataset can be reused to assess the ability of other wavelengths and power levels to kill microorganisms as well as improve procedures for treating endodontic infections and inflammation in root canals.
Background: All-trans-retinoyl beta-D-glucuronide, a water-soluble glucuronic acid conjugate of a... more Background: All-trans-retinoyl beta-D-glucuronide, a water-soluble glucuronic acid conjugate of all-trans-retinoic acid, has demonstrated high biological activity and low toxicity in most in vitro and in vivo models. Since the reparative effects of retinoids on epithelium are well-known, our aim was to study the effect(s) of intravenously-administered all-trans-retinoyl beta-D-glucuronide in lambs with chronic bacterial bronchopneumonia. Material/methods: Two groups of lambs were inoculated intrabronchially with either pyrogen-free saline or Mannheimia haemolytica. Thirty-three days later, lambs were injected four times at five-day intervals with 2 mL of 116 mM all-trans-retinoyl beta-D-glucuronide (6.0-9.3 mmol/kg or 2.86-4.42 mg/kg animal body weight) in dimethyl sulfoxide, or dimethyl sulfoxide alone. Animal behavior and signs of clinical illness were logged daily. Lung and liver samples were assessed for histopathology, while serum and liver samples were extracted for retinoids and analyzed by reversed-phase gradient high-performance liquid chromatography. Results: Repeated injections of highly concentrated all-trans-retinoyl beta-D-glucuronide did not cause changes in appetite, activity or other behaviors nor did it cause histologic lesions in liver and lung. However, it had no effect on resolution of lung lesions resultant of chronic Mannheimia haemolytica bronchopneumonia. Conclusions: Repeated intravenous administration of high amounts of all-trans-retinoyl beta-D-glucuronide to lambs did not elicit signs of gross or microscopic toxicity. However, administering all-trans-retinoyl beta-D-glucuronide too late in the progression of bacterial pneumonia is thought to be the main reason for its lack of effect in this model. A retinoid lactone derivative was detected in sheep serum and liver several days after treatment.
Medical science monitor : international medical journal of experimental and clinical research, 2002
All-trans-retinoyl beta-D-glucuronide, a water-soluble glucuronic acid conjugate of all-trans-ret... more All-trans-retinoyl beta-D-glucuronide, a water-soluble glucuronic acid conjugate of all-trans-retinoic acid, has demonstrated high biological activity and low toxicity in most in vitro and in vivo models. Since the reparative effects of retinoids on epithelium are well-known, our aim was to study the effect(s) of intravenously-administered all-trans-retinoyl beta-D-glucuronide in lambs with chronic bacterial bronchopneumonia. Two groups of lambs were inoculated intrabronchially with either pyrogen-free saline or Mannheimia haemolytica. Thirty-three days later, lambs were injected four times at five-day intervals with 2 mL of 116 mM all-trans-retinoyl beta-D-glucuronide (6.0-9.3 mmol/kg or 2.86-4.42 mg/kg animal body weight) in dimethyl sulfoxide, or dimethyl sulfoxide alone. Animal behavior and signs of clinical illness were logged daily. Lung and liver samples were assessed for histopathology, while serum and liver samples were extracted for retinoids and analyzed by reversed-phase...
Treatment of root canals with ultraviolet C (UVC) light-emitting diodes (LED) is a promising adju... more Treatment of root canals with ultraviolet C (UVC) light-emitting diodes (LED) is a promising adjunct to current root canal disinfection procedures that are essential for preserving the health and the healing of periapical tissues. Here, we hypothesized that UVC LED treatment would kill microorganisms associated with endodontic infections and kill microorganisms in instrumented root canal models. Three different UVC LED units were constructed. One unit emitted 265 nm from a 12 mW LED, one unit emitted 265 nm from a 22.5 mW LED, and one unit emitted 280 nm from an 8 mW LED. Power levels emitted from the three units were measured without and with fiberoptic (FO) filaments using an energy meter (PM100D, Thorlabs, Inc., Newton, NJ USA) with a standard photodiode sensor (S120VC, 200-1100 nm, 50 mW; Thorlabs, Newton, NJ USA) for 30 seconds. The raw data from the energy meter readings for these 3 units is listed in Table 1. This includes the wavelength the meter used to record the data for each unit (Wavelength read at (nm)), the distance the unit was from the sensor (Distance from sensor (mm)), the minimum power value recorded from the meter (Minimum value (µW)), the maximum power value recorded from the meter (Maximum value (µW)), the mean value recorded from the meter (Mean value (µW)), the standard deviation of the mean value recorded from the meter (Standard deviation (µW)), the number of meter readings (Number of readings (n)), and the time of recording (Time (sec)). The raw data from the antimicrobial activities of these 3 units on Candida albicans ATCC 64124, Enterococcus faecalis ATCC 29212, Streptococcus sanguinis ATCC 10556, Staphylococcus aureus ATCC 29213, and methicillin-resistant S. aureus (MRSA #7) treated on discs in laboratory assays is listed in Table 2. This includes the length of time each microorganism was treated (Group (seconds)), the UVC LED unit used to treat each microorganism (UVC LED treatment), the replication number (Replication), the microorganism treated (Microorganisms), and the numbe [...]
Although some vaccines and antibiotics have been very effective in preventing and treating respir... more Although some vaccines and antibiotics have been very effective in preventing and treating respiratory disease, they have not been fully satisfactory. Recently, components of the innate immune system have been increasingly appreciated for their role in host defense against microbial pathogens. These molecules include lysozyme and lactoferrin, but recent work in cattle, sheep, man and other species have identified new classes of peptides expressed by respiratory epithelial cells that have potent microcidal activity in nanogram quantities. These peptides, termed antimicrobial peptides (AMP), include defensins, cathelicidins and anionic peptides. Some are expressed continuously whereas expression of others is stimulated by infection/inflammation. In calves, we have found that two AMP from the defensin family, tracheal antimicrobial peptide (TAP) and lingual antimicrobial peptide (LAP), are expressed in the newborn and increased in response to Mannheimia (Pasteurella) haemolytica infection. In contrast, sheep beta defensin-1 (SBD-1) is not induced by infection and, in fact, appears decreased during viral infection with paramyxovirus-3 (PI-3). Decreased SBD-1 by Pl-3 may increase the lung's susceptibility to secondary infections or re-infections. Other innate defense molecules include proteins released with lung surfactant. Surfactant protein A and D (SAD) can opsonize and aggregate respiratory syncytial virus (RSV) and activate alveolar macrophages. Preliminary work suggests that chronic bacterial infections result in reduced SpA and SpD expression and we are currently assessing SAD expression in response to Pl-3 and RSV. A long-term goal of our work is to identify ways to up-regulate expression of AMP and/or surfactant proteins in the neonate and at times of stress in older animals in order to r educe microbial colonization. Other investigators are seeking ways to utilize AMP's as a new class of antibiotics .
American Journal of Veterinary Research, May 1, 2001
Objectives-To determine effects of selectin inhibitor TBC1269 on neutrophil infiltration, and neu... more Objectives-To determine effects of selectin inhibitor TBC1269 on neutrophil infiltration, and neutrophilassociated injury during pneumonia induced by Mannheimia haemolytica and concentration of antimicrobial anionic peptide (AAP) in bronchoalveolar lavage fluid (BALF) as well as antimicrobial activity of BALF from healthy (control) neonatal calves, neonatal calves with M haemolytica-induced pneumonia, neonatal calves with prior treatment with TBC1269, and adult cattle. Animals-Eighteen 1-to 3-day-old calves and 9 adult cattle. Procedure-Calves were inoculated with M haemolytica or pyrogen-free saline (0.14M NaCl) solution into the right cranial lung lobe, and BALF was collected 2 or 6 hours after inoculation. Thirty minutes before and 2 hours after inoculation, 4 calves received TBC1269. The BALF collected from 9 adult cattle was used for comparison of BALF AAP concentration and antimicrobial activity. Protein concentration and neutrophil differential percentage and degeneration in BALF were determined. An ELISA and killing assay were used to determine BALF AAP concentration and antimicrobial activity, respectively. Results-Total protein concentration was significantly decreased in BALF from calves receiving TBC1269. Similar concentrations of AAP were detected in BALF from all calves, which were 3-fold higher than those in BALF from adult cattle. However, BALF from neonates had little or no anti-M haemolytica activity. Conclusions and Clinical Relevance-These results suggest that TBC1269 decreases pulmonary tissue injury in neonatal calves infected with M haemolytica. Although AAP is detectable in neonatal BALF at 3 times the concentration detected in adult BALF, neonatal BALF lacks antimicrobial activity for M haemolytica.
FREE-LIVING AND HOST-ASSOCIATED PROTOZOA AS TRAINING CAMPS FOR INTRACELLULAR PATHOGENS Steve A. C... more FREE-LIVING AND HOST-ASSOCIATED PROTOZOA AS TRAINING CAMPS FOR INTRACELLULAR PATHOGENS Steve A. Carlson, Sharon K. Franklin, and Mark A. Rasmussen 9 In 1976, a group of American Legion members gathered in Philadelphia, PA, for a convention. ...
Ovine progressive pneumonia is caused by a lentivirus of known infectivity only for sheep and goa... more Ovine progressive pneumonia is caused by a lentivirus of known infectivity only for sheep and goats. Virus susceptibility of 11 other species of animals was examined. Species included cattle, chickens, deer, dogs, goats, hamsters, horses, mice, pigs, rabbits, and rats. Of these species, only goats and rabbits could be experimentally infected with the virus. The infection in rabbits was acute, and virus did not persist or induce antibody production as it does in sheep and goats. Sera obtained from several people working in close contact with the virus and from several wild species, with unknown exposure history, were tested for antibodies to viral antigens. All results were negative. Knowledge of the host range of this virus is important for scientific studies and for virus eradication programs.
Cell-mediated immune mechanisms may play a role in the pathogenesis and prevention of pneumonia i... more Cell-mediated immune mechanisms may play a role in the pathogenesis and prevention of pneumonia in cattle caused by Pasteurella haemolytica serotype A1. To determine the circumstances required to stimulate and identify cell-mediated immune responses, calves were vaccinated with a commercial P. haemolytica bacterin or a live commercial P. haemolytica vaccine, or were infected intratracheally with virulent P. haemolytica. All calves were challenge-exposed intratracheally with P. haemolytica 31 d after vaccination or prior infection. Peripheral blood mononuclear cells and mediastinal and superficial cervical lymph node cells were stimulated with antigens prepared from P. haemolytica to evaluate in vitro proliferative responses and gamma-interferon production as measures of cell-mediated immunity. Strong proliferative responses and gamma-interferon production were detected in lymph node cells from calves vaccinated with the live vaccine and from infected calves, especially in response to stimulation with an outer membrane protein preparation from P. haemolytica. Greater proliferative responses and gamma-interferon production were associated with the lymph node nearer the site of bacterin administration (superficial cervical lymph node) or the site of infection (mediastinal lymph node), whereas greater proliferative responses and gamma-interferon production were associated with the more distant lymph node (mediastinal lymph node) in calves vaccinated with the live vaccine. Neither proliferative responses nor gamma-interferon production were detected in peripheral blood mononuclear cells from calves that were vaccinated for or infected with P. haemolytica. Antileukotoxin antibody titers were determined by a serum neutralization assay, and protection against pneumonic lesions was more closely correlated with antileukotoxin antibody responses than with lymphocyte proliferation or gamma-interferon responses.
A field trial to evaluate a whole cell vaccine for the prevention of caseous lymphadenitis (CLA) ... more A field trial to evaluate a whole cell vaccine for the prevention of caseous lymphadenitis (CLA) in sheep and goats was performed in one goat herd and one sheep flock over a period of three years. In goats, there was a nonstatistically significant trend for fewer cases of CLA in the vaccinated animals compared to the controls. In sheep, from six months to 36 months postinitial vaccination, the proportion of vaccinated sheep that developed CLA was significantly less (p less than 0.05) than in the control sheep. The antibody titers to Corynebacterium pseudotuberculosis as detected by microagglutination assay were significantly different (p less than 0.0001) at all times except at the initial vaccination. Swellings occurred at the vaccination site at an incidence level of 29.6% in goats and 34.1% in sheep. The vaccine appeared to be efficacious in reducing the proportion of sheep that developed CLA when challenged naturally in a field situation.
In Alberta, caseous lymphadenitis (CLA) is one of the leading causes of lamb and mutton carcass c... more In Alberta, caseous lymphadenitis (CLA) is one of the leading causes of lamb and mutton carcass condemnation. In this study, serologic results confirmed a high (50-94%) incidence of exposure to Corynebacterium pseudotuberculosis, the causative agent of CLA, in mature, unvaccinated sheep in southern Alberta. To assess the efficacy and impact of vaccination with 2 commercial (Glanvac-6 and Case-Vac) and 1 experimental (WC+ MDP-GDP) CLA vaccines, a series of 3 field trials in 3249 ewes and lambs was conducted in affected flocks from 1992-1996. Efficacy was assessed from the serological response to vaccination, prevalence and size of injection site reactions by treatment, and the incidence of CLA abscesses. Overall, agglutinating antibody titres to C. pseudotuberculosis in lambs vaccinated with WC+MDP-GDP and Case-Vac remained significantly elevated above nonvaccinated control lambs for the 12 mo period after the initial vaccination. Lambs vaccinated with the WC/MDP-GDP maintained higher titres (P < 0.06) than those vaccinated with Case-Vac for the period from 6 to 12 mo after vaccination. Agglutinating antibody titres for lambs vaccinated with Glanvac did not differ from those of controls at any point during the 12 mo period after vaccination. The number of injection site reactions was elevated in lambs vaccinated with Glanvac as compared to those vaccinated with WC+MDP-GDP but the size of injection site reactions did not significantly differ. Sheep vaccinated with WC+ MDP-GDP also had a reduced incidence of putative CLA abscesses, although confirmation of the presence of C. pseudotuberculosis was only successful in a small number of instances.
Almost two decades after identification of the CFTR gene, we lack answers to many questions about... more Almost two decades after identification of the CFTR gene, we lack answers to many questions about the pathogenesis of cystic fibrosis (CF), and it remains a lethal disease. Mice with a disrupted CFTR gene have greatly facilitated CF studies, but they fail to develop the characteristic pancreatic, lung, intestinal, liver, and other CF manifestations. Therefore, we produced pigs with a targeted disruption of both CFTR alleles. These animals exhibited defective chloride transport. They also developed meconium ileus, exocrine pancreatic destruction, and focal biliary cirrhosis, replicating abnormalities seen in newborn patients with CF. This swine model may provide opportunities to address persistent questions about CF pathogenesis and accelerate discovery of treatments and preventions. Understanding human disease often requires animal models. Mice have been the overwhelming species of choice because methods for specifically altering their genome have
Although some vaccines and antibiotics have been very effective in preventing and treating respir... more Although some vaccines and antibiotics have been very effective in preventing and treating respiratory disease, they have not been fully satisfactory. Recently, components of the innate immune system have been increasingly appreciated for their role in host defense against microbial pathogens. These molecules include lysozyme and lactoferrin, but recent work in cattle, sheep, man and other species have identified new classes of peptides expressed by respiratory epithelial cells that have potent microcidal activity in nanogram quantities. These peptides, termed antimicrobial peptides (AMP), include defensins, cathelicidins and anionic peptides. Some are expressed continuously whereas expression of others is stimulated by infection/inflammation. In calves, we have found that two AMP from the defensin family, tracheal antimicrobial peptide (TAP) and lingual antimicrobial peptide (LAP), are expressed in the newborn and increased in response to Mannheimia (Pasteurella) haemolytica infection. In contrast, sheep beta defensin-1 (SBD-1) is not induced by infection and, in fact , appears decreased during viral infection with paramyxovirus-3 (Pl-3). Decreased SBD-1 by Pl-3 may increase the lung's susceptibility to secondary infections or re-infections. Other innate defense molecules include proteins released with lung surfactant. Surfactant protein A and D (SAD) can opsonize and aggregate respiratory syncytial virus (RSV) and activate alveolar macrophages. Preliminary work suggests that chronic bacterial infections result in reduced SpA and SpD expression and we are currently assessing SAD expression in response to PI-3 and RSV. A long-term goal of our work is to identify ways to up-regulate expression of AMP and/or surfactant proteins in the neonate and at times of stress in older animals in order to reduce microbial colonization. Other investigators are seeking ways to utilize AMP's as a new class of antibiotics .
CXCL10 (IP-10) is a small 10 kDa chemokine with antimicrobial activity. It is induced by IFN-γ, c... more CXCL10 (IP-10) is a small 10 kDa chemokine with antimicrobial activity. It is induced by IFN-γ, chemoattracts mononuclear cells, and promotes adhesion of T cells. Recently, we detected CXCL10 on the surface of the skin and in the oral cavity. In the current study, we used broth microdilution and radial diffusion assays to show that CXCL10 inhibits the growth of Escherichia coli, Staphylococcus aureus, Corynebacterium jeikeium, Corynebacterium striatum, and Candida albicans HMV4C, but not Corynebacterium bovis, Streptococcus mutans, Streptococcus mitis, Streptococcus sanguinis, Fusobacterium nucleatum, Aggregatibacter actinomycetemcomitans, Poryphromonas gingivalis, or C. albicans ATCC 64124. The reason for the selective antimicrobial activity is not yet known. However, antimicrobial activity of CXCL10 may be related to its composition and structure, as a
Ultraviolet C (UVC) light emitting diode (LED) can kill the endodontic pathogen Enterococcus faec... more Ultraviolet C (UVC) light emitting diode (LED) can kill the endodontic pathogen Enterococcus faecalis and has the potential to kill other oral microorganisms associated with endodontic infections. This same bacteriocidal device shows great promise in the stimulation of periapical healing and pain reduction resulting from inflammation in root canals. Previously, we found that 255 nm UVC LED killed E. faecalis and induced the production of cellular biomarkers in HEPM cells and gingival fibroblasts (Morio et al., 2019). Here, we extend those findings and hypothesize that UVC LED at other wavelengths and power levels kill microorganisms associated with root canal infections. Units emitting UVC LED at 265 nm (12 mW), 265 nm (22.5 mW), and 280 nm (8 mW) wavelenths were assembled and the energy levels of their emissions were measured. The energy doses in millijoules (mJ) were calculated from the power readings of the meter (µW) × time of exposure (seconds). Ex vivo models of root canals were prepared in extracted, instrumented, single canal human premolars. Five cultures of microorganisms were treated with 265 nm (12 mW), 265 nm (22.5 mW), or 280 nm (8 mW) UVC LED on discs in laboratory assays and 4 cultures of microorganisms were treated with 265 nm (22.5 mW) UVC LED in root canals of extracted, instrumented teeth. After UVC LED treatment, all microorganisms were cultivated on microbiological media. Colony forming units (CFU) of viable microorganisms treated with UVC LED were counted and compared with those of viable microorganisms not treated with UVC LED as controls. Tukey's Honestly Significant Difference was used to determine statistical significances (0.05). Units emitting UVC LED at 265 nm (12 mW), 265 nm (22.5 mW), and 280 nm (8 mW) killed Candida albicans, Staphylococcus aureus, methicillin-resistant S. aureus (MRSA), E. faecalis, and Streptococcus sanguinis after 30-90 seconds of exposure in laboratory assays (p < 0.05). Microbial killing differed among treatment times, UVC LED wavelengths, power levels of each unit, and specific microorganism. The unit emitting UVC LED at 265 nm (22.5 mW) killed C. albicans, S. aureus, MRSA, and E. faecalis in 30 s in root canals of extracted, instrumented teeth (p < 0.05). This dataset can be reused to assess the ability of other wavelengths and power levels to kill microorganisms as well as improve procedures for treating endodontic infections and inflammation in root canals.
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Papers by Kim Brogden