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1
Hassan, H., Gayatri Jaishankar, and Demetrio Macariola. "The "Non" Whooping Cough." Digital Commons @ East Tennessee State University, 2020. https://dc.etsu.edu/etsu-works/8861.
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Torvaldsen, Siranda. "The epidemiology and prevention of pertussis in Australia." University of Sydney. Paediatrics and Child Health, 2001. http://hdl.handle.net/2123/808.
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Pertussis (whooping cough) remains an important public health problem in Australia. Although mortality and morbidity from pertussis declined dramatically following the introduction of mass vaccination programs in 1953, the level of morbidity remains unacceptably high for a vaccine-preventable disease. Aims and methods The primary aims of this thesis were (i) to ascertain the epidemiology of pertussis in Australia between 1993 and 2000 by analysing and interpreting sources of routinely collected data on pertussis; and (ii) to examine the effectiveness of vaccination against pertussis in a number of ways. Data from three primary national sources (notifications of disease, hospitalisations for pertussis and death certificates) were used to examine the burden from pertussis in Australia over these eight years. Analyses included the age distribution of cases, temporal and geographic trends, comparisons of notification and hospitalisation data, and the impact of differences in the method of diagnosis of notified cases between years and age groups. In addition to analyses at the national level using data from the national databases, further detailed analyses were undertaken at the State level for New South Wales (NSW), the most populous Australian State. Pertussis vaccine coverage was estimated using data from the recently established Australian Childhood Immunisation Register (ACIR); these data were also used to track the transition from whole-cell to acellular pertussis vaccines. The different types of studies used to evaluate vaccine effectiveness were reviewed, and a method suitable for ongoing estimation of vaccine effectiveness in Australia was developed. This was then applied to the NSW data, to determine the effectiveness of pertussis vaccination in this State. Main findings The annual notification rate for pertussis in Australia ranged from 23�59 per 100 000 population over the eight years. Infants had the highest notification and hospitalisation rates in Australia � they accounted for 5percent of notifications, 61percent of hospitalisations and 100percent of deaths. Age-specific notification and hospitalisation rates in children aged less than two years strongly suggested a protective effect of vaccination, with the greatest reduction in rate coinciding with eligibility to receive a second dose of pertussis vaccine at four months of age. Notification rates among 5�9 year olds progressively decreased in successive age cohorts, consistent with an effect of the introduction in 1994 of a pertussis vaccine booster for preschool-aged children. Although adults (persons aged 15 years or more) accounted for half the notifications, they had the lowest notification rate. The highest numbers of pertussis notifications were in 1997, when most jurisdictions experienced an epidemic. Notification and hospitalisation rates varied across the States and Territories and also across smaller geographic regions in NSW. Areas and years with high notification rates tended to also have high hospitalisation rates, suggesting that trends in notifications reflected trends in incidence. The number of infant hospitalisations in NSW between July 1993 and June 1999 exceeded the number of notifications by 32percent, highlighting the extent of under-notification. Overall, and particularly amongst those aged more than 12 months, the majority of cases notified in NSW were based on the results of serological tests. The proportion diagnosed by culture of the organism was greatest in infants; the proportion diagnosed by serological tests increased with age. There was no evidence that the use of serology had increased since 1994 in NSW, hence changes in notification rates after this time are unlikely to be attributable to increased use of serological diagnosis. ACIR records indicated that in December 2000, 92percent of one-year-old children had received three doses of diphtheria-tetanus-pertussis (DTP) vaccine and 90percent of two-year-olds had received four doses. Vaccine coverage varied by jurisdiction. Since 1997, there was an increased use of DTP vaccines containing acellular pertussis components with a corresponding decrease in the use of vaccines containing whole-cell components. In 2000, almost all DTP vaccines administered contained acellular pertussis components. The results of the vaccine effectiveness study showed that pertussis vaccination was highly effective at preventing pertussis in NSW children, as measured by notified cases. Vaccine effectiveness was highest (91percent) in the youngest age group (8�23 months) and lowest (78percent) in the oldest age group (9�13 years). The screening method has not previously been used to estimate pertussis vaccine effectiveness in Australia. Conclusions This thesis demonstrates the value of integrating varied data sources in estimating the disease burden from pertussis. The data presented here show that the disease burden is substantial in all age groups, despite high levels of vaccine coverage in infants and children. This problem of disease control does not appear to be due to lack of vaccine effectiveness, but there is evidence of waning immunity over time. The analyses presented here form a basis for the ongoing monitoring of trends in pertussis epidemiology following the replacement of whole-cell by acellular pertussis vaccines, and will assist consideration of the need for additional booster doses in adolescents and adults.
3
Piyawong, Wirawan. "Spatio-temporal numerical modelling of whooping cough dynamics." Thesis, Brunel University, 2001. http://bura.brunel.ac.uk/handle/2438/6626.
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The SIR (Susceptible/Infectious/Recovered) whooping cough model involving nonlinear ordinary differential equations is studied and extended to incorporate (i) diffusion (ii) convection and (iii) diffusion-convection in one-space dimension. Firstand second-order finite-difference methods are developed to obtained the numerical solutions of the ordinary differential equations. Though implicit in nature, with the resulting improvements in stability, the methods are applied explicitly. The proposed methods are economical and reliable in comparison to classical numerical methods. When extended to the numerical solutions of the partial differential equations, the solutions are found by solving a system of linear algebraic equations at each time step, as opposed to solving a non-linear system, which often happens when solving non-linear partial differential equations.
4
Johnston, I. D. A. "The current severity and longterm sequelae of whooping cough." Thesis, University of Cambridge, 1985. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.605661.
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Cheung, Yung-yan Terence. "Whooping cough : are we seeing the reemergence of the infection in Hong Kong? /." Click to view the E-thesis via HKUTO, 2005. http://sunzi.lib.hku.hk/hkuto/record/b39724049.
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McKenna, Philip Rood. "Winging it : a bold step toward the whooping crane's return." Thesis, Massachusetts Institute of Technology, 2006. http://hdl.handle.net/1721.1/39445.
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Thesis (S.M. in Science Writing)--Massachusetts Institute of Technology, Dept. of Humanities, Graduate Program in Science Writing, 2006.Includes bibliographical references (leaves 41-42).
Since the fall of 2001, biologists have taught endangered whooping cranes how to migrate over a once-lost course stretching from the wetlands of central Wisconsin to the mud flats of Florida's Gulf Coast. Wildlife biologists did this through an unusual method of reintroduction: training the endangered birds to follow behind ultralight airplanes for the entire 1,200-mile journey. The technique is highly invasive and expensive, but by the summer of 2005, it had established the first population of whooping cranes migrating east of the Mississippi in more than one hundred years. To supplement these ultralight-led migrations, crane biologists tried a new approach in the fall of 2005. Biologists with the International Crane Foundation of Baraboo, Wisconsin, and the U.S. Fish and Wildlife Service released four captive-bred whooping cranes directly into the wild. Biologists hoped that there were enough graduates of the ultralight program already making the migration for a few first timers to simply follow the older birds south. But no one knew if this bold new experiment, which relied entirely on the young birds following older non-related birds, would work. This thesis follows a year in the life of Maya, Poe, Waldo and Jumblies-the first four "Direct Autumn Release" birds.
(cont.) The story begins with their parent's artificial insemination in the spring of 2005, describes their last-minute Thanksgiving-Day departure, and follows their successful southern migrations through Tennessee and Florida. The thesis relates the concerns of the biologists, who spent countless hours raising and tracking these birds. It also recounts historic episodes in the 80-year ongoing effort to save Grus Americana, the whooping crane, while providing a larger significance for why the conservation of biodiversity is needed now more than ever.
by Philip Rood McKenna.
S.M.in Science Writing
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Gil, de Weir Karine. "Whooping crane (Grus americana) demography and environmental factors in a population growth simulation model." Texas A&M University, 2005. http://hdl.handle.net/1969.1/3778.
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The Whooping Crane (Grus americana) is among North AmericaÂs most
charismatic species. Between 1938 and 2004, the population that migrates between
Aransas National Wildlife Refuge (ANWR) and Wood Buffalo National Park (WBNP),
grew from 18 to 217 individuals. The recovery plan objective for this endangered
species is to downlist the population in 2035, but this requires interpretive assessment of
population responses to environmental factors over the long term. I analyzed 27 years of
banding data, 37 years of nest monitoring data, and 20 years of winter reports to estimate
age-specific mortality and fecundity rates. The resulting life table yielded an intrinsic
rate of increase (r) of 0.14/y, a net reproductive rate (Ro) of 6.4/y, and a mean length of
a generation (G) of 13y.
Path analysis of environmental factors, demographic variables (natality and
mortality), and the finite rate of population increase (lambda) showed that annual
mortality, temperatures from the ANWR, WBNP and at a migration stop-over in Nebraska, and pond water depth were good predictors of lambda variability. However,
other environmental factors were significantly correlated: at ANWR, October- March
temperature (extreme minimum and maximum), December temperature (mean and
extreme minimum), November-January precipitation, and September-March freshwater
inflow; at WBNP, March-September precipitation, March-May temperature, and
temperatures during the September - October fall migration. The Pacific Decadal
Oscillation (PDO) affected lambda indirectly through environmental factors in Nebraska
and ANWR.
I graphically analyzed relevant data trends from 1967 to 2004 to identify the
relation between phases of PDO and environmental and demographic variables. During
PDO cold phases, a synchronization of Âextreme environmental values was observed
from the different regions; during warm phases extreme environmental values were
scattered. Most periods of Whooping Crane population decline happened during cold
phases.
I developed a compartment model to represent Whooping Crane population
dynamics utilizing the new data on survivorship and fecundity from banded birds. The
model was capable of simulating historical population trends with adjustments in brood
success and egg mortality. The model will allow future studies to test population
responses to various environmental scenarios at the WBNP, during fall and spring
migrations, and at the ANWR.
8
Srikannathasan, Velupillai. "Biochemical and structural analysis of Bordetella pertussis lipopolysaccharide A biosynthetic pathway enzymes." Thesis, University of East Anglia, 2002. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.273576.
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Al-Fellah, Giamal Nouri. "Inactivation of Bordetella pertussis by rat lung lavage fluids (LLF)." Thesis, University of Glasgow, 1997. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.263427.
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Brotherston, Christopher. "Interaction of Bordetella pertussis adenylate cyclase toxin with target cells." Thesis, University of Glasgow, 1997. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.263429.
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MacDonald-Fyall, Julia. "The protective and immunomodulatory properties of Bordetella pertussis adenylate cyclase toxin." Thesis, University of Glasgow, 2002. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.248267.
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Garrod, Tracey. "Partial purification and characterisation of Bordetella bronchiseptica dermonecrotic toxin." Thesis, University of Reading, 1994. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.239514.
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Funnell, Simon Gordon Paul. "Mechanisms of colonisation of mammalian tissues by Bordetella pertussis." Thesis, Open University, 1994. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.239726.
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Advani, Abdolreza. "Epidemiological characterisation of Bordatella pertussis in Sweden, 1970-2004 /." Stockholm, 2007. http://diss.kib.ki.se/2007/978-91-7357-052-7/.
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Gorringe, A. R. "The effects of growth conditions on the expression of virulence determinants of Bordetella pertussis." Thesis, University of Southampton, 1988. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.235294.
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Berry, P. R. "Production of monoclonal antibodies to Bordetella pertussis as a means of identifying protective antigens." Thesis, University of Reading, 1987. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.376819.
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Li, Jing-Li. "A molecular study of virulence factors of Bordetella species." Thesis, Open University, 1990. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.303193.
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A 1kb HinfI DNA fragment, containing a repeat DNA sequence element was isolated from a Bordetella pertussis BP348 cosmid gene bank. This repeat DNA sequence has subsequently been named IS148. The insertion sequence was demonstrated by have a high copy number only in B.pertissus and to be absent in Bordetella parapertussis and Bordetella bronchiseptica. Using a restriction fragment of IS148 labelled with radioactivity as a probe in DNA or whole cell dot blots between 10-100 cells could be detected or from 100pb-1ng DNA. Furthermore, a pair of oligonucleotide primers was synthesised corresponding to a central region of IS148 that could generate a 242bp fragment upon PCR amplification. The use of PCR amplification with specific primers allows the detection of 0.5pg of B.pertussis chromosomal DNA and as little as one B.pertussis cell. The prn genes encoding the outer membrane P.70 and P.68 pertactin from B.parapertussis and B.bronchiseptica have been cloned, sequenced and expressed in Escherichia coli. Analysis of the DNA sequences reveal that the genes have open reading frames capable of encoding proteins with molecular weights of 95,177 (P.95, B.parapertussis) and 93,996 (P.94, B.bronchiseptica). These precursors molecular are processed to form the P.70 and P.68 antigens on the surface of B.parapertussis and B.bronchiseptica respectively. Heterologous expression of the full-length gene encoding P.95 and P.94 in E.coli result in similar processing, with the P.70 and P.68 antigens targetted to the bacterial outer membrane. Comparison of P.95, P.94 and P.93, encoding homologous proteins from B.parapertussis, B.bronchiseptica and B.pertussis, shows a high degree (> 90%) of homology. The major differences between all three proteins occur in the number of repeat of the two families (Gly-Gly-Xaa-Xaa-Pro)n and (Pro-Gln-Pro)n of reiterated sequence motifs.
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Cheung, Yung-yan Terence, and 張勇仁. "Whooping cough: are we seeing the reemergenceof the infection in Hong Kong?" Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2005. http://hub.hku.hk/bib/B39724049.
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White, Jennifer L. "Management of captive whooping cranes (Grus americana ) to improve breeding behaviour and success." Thesis, National Library of Canada = Bibliothèque nationale du Canada, 2000. http://www.collectionscanada.ca/obj/s4/f2/dsk1/tape4/PQDD_0029/MQ66980.pdf.
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Pillay, Victoria. "A systematic review of the symptomatic treatment of the cough in whooping cough." Master's thesis, University of Cape Town, 2002. http://hdl.handle.net/11427/8789.
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Includes bibliographical references.Background: There are between 20 - 40 million cases of whooping cough annually world-wide, 90% of which occur in developing countries, resulting in an estimated 200 - 300 O00 fatalities each year. Much of the morbidity is due to the paroxysmal cough. Corticosteroids, salbutamol (a β₂- adrenergic stimulant), and pertussis-specific immunoglobulin have been proposed as standard treatment for the cough. Antihistamines have also been administered. No systematic review of the effectiveness of any of these interventions or others has been performed. Objective: in this systematic review we aim to assess the effectiveness of interventions used to reduce the severity of the coughing paroxysms in whooping cough in children and adults. Selection criteria: Randomised and quasi randomised controlled trials of any intervention that reduced the severity of the coughing paroxysms in whooping cough; excluding antibiotics and vaccines. Study selection: All interventions aimed at reducing the severity of the coughing paroxysms in children or adults with whooping cough with any of the following outcome measures met inclusion criteria for the review; i) frequency of paroxysms of coughing (primary outcome), ii) frequency of vomiting, iii) frequency of whoop, iv) frequency of cyanotic spells, v) development of serious complications, vi) mortality from any cause, vii) side effects due to medication, viii) admission to hospital, and ix) duration of hospital stay. Search strategy: We searched the Cochrane Controlled Trials register, Acute Respiratory infectious Disease Group Specialised Trials register, MEDLINE, LILACS, scanned reference lists of identified trials, contacted authors of identified trials and the relevant pharmaceutical companies. Data collection and analysis: Studies were selected, quality assessed and data extracted by two reviewers independently. Results: Nine studies satisfied the inclusion criteria but four had insufficient data for further meta-analysis of our pre-specified outcomes. Studies were old and poorly reported. The largest study had a total sample size of 49 and the smallest study nine. All studies were performed in industrialised settings. Eligible studies assessed diphenhyramine, pertussis immunoglobulin, dexamethasone and salbutamol. No statistically significant benefit was found for any of the interventions. Diphenhydramine was associated with a mean increase of 1.90 coughing spells per 24 hours [95%Cl - 4.66; 8.46] and pertussis immunoglobulin a mean decrease in hospital stay of 0.70 days [95% Cl -3.79; 2.39]. and a mean reduction of 3.10 whoops per 24 hours [95% CI - 6.22; 0.02]. Dexamethasone resulted in a mean decrease in hospital stay of 3.45 days [95% Cl - 15.34; 8.44] and salbutamol in a weighted mean decrease in coughing paroxysms of 0.95 per 24 hours [95% Cl - 6.21; 4.31]. Reviewers' conclusion: Although assessments have been performed on a whole range of interventions, including diphenhyramine, pertussis immunoglobulin, dexamethasone and salbutamol, insufficient evidence exists to draw conclusions about the effects of any of them.
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Sidey, Fiona M. "Metabolic effects of Bordetella pertussis." Thesis, University of Strathclyde, 1987. http://oleg.lib.strath.ac.uk:80/R/?func=dbin-jump-full&object_id=20352.
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The present work confirmed that B. pertussis infection or pertussis toxin produce hypoglycaemia in mice. The hypoglycaemia was associated with hyperinsulinaemia, and both were abolished by destruction of the pancreatic β cells with alloxan. Impaired glucose counterregulatory mechanisms may also contribute to pertussis-induced hypoglycaemia, as the hypoglycaemic action of insulin was prolonged in pertussis infected mice. On the other hand, impaired responsiveness to lower doses of insulin was found. Pertussis-induced hyperinsulinaemia had two components. First, the increase in serum insulin in response to food intake was both greater and more prolonged in pertussis-infected mice. Second, infected or pertussis toxin-treated animals, unlike controls, showed a marked increase in serum insulin in response to certain stresses, such as ether, histamine, anoxia and 2-deoxyglucose. However, other stresses (LPS, cold and hypoxia) did not cause hyperinsulinaemia in pertussis infected mice. Stress-induced hyperinsulinaemia was also seen in normal mice receiving the a2- adrenoceptor blocking drug idazoxan. Stress-induced hyperinsulinaemia in a2 adrenoceptor blocked mice, but not in pertussis-treated mice, was prevented by β adrenoceptor blockade using propranolol. Adrenal demedullation or ganglionic blockade (using hexamethonium) in normal mice also allowed stress induced hyperinsulinaemia. Thus, adrenal medullary catecholamines may normally serve to prevent stress induced hyperinsulinaemia, which becomes unmasked when they are absent or when their action is prevented. Stress-induced hyperinsulinaemia in pertussis treated mice was unlikely to involve autonomic, cholinergic oropioid mechanisms as it was not blocked by hexamethonium, atropine or naloxone. Human infants with pertussis showed no hypoglycaemia compared with non-pertussis controls, although their plasma insulin concentrations were slightly but significantly raised. It remains possible that hyperinsulinaemia with resultant profound hypoglycaemia might occur in susceptible patients following exposure to pertussis-toxin (either during the disease or following vaccination).
22
Khan, Farhat Mirza. "Effects of toxoiding agents on protective antigens of Bordetella pertussis and on other proteins." Thesis, University of Glasgow, 1997. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.360122.
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Lund, Sarah Jane. "Virulence of Bordetella parapertussis : a comparison of ovine and human isolates." Thesis, University of Glasgow, 1999. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.314103.
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Phillips, Linda Jane. "Immunization against Bordetella pertussis." Thesis, Kansas State University, 1985. http://hdl.handle.net/2097/9871.
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Fry, Scott Robert. "The development and evaluation of DNA vaccines against whooping cough using a murine respiratory model of infection." University of Southern Queensland, Faculty of Sciences, 2006. http://eprints.usq.edu.au/archive/00004787/.
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In this study, a suite of single antigen DNA vaccines, combination DNA vaccines and dual modality vaccines, were developed and evaluated for their potential to induce humoral and cell-mediated immune responses, and protective efficacy against Bordetella pertussis, the aetiological agent of whooping cough, using the mouse respiratory challenge model.
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Cahill, Edward Sean. "Antigen delivery systems for nasal immunisation against B. pertussis." Thesis, University of Nottingham, 1996. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.321455.
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Castillo, María Esther, Carlos Bada, Aguila Olguita Del, Helasvuo Verónica Petrozzi, Ore Verónica Casabona, Isabel Reyes, and Valle Mendoza Juana Mercedes Del. "Detection of Bordetella pertussis using a PCR test in infants younger 3 than one year old hospitalized with whooping cough in five 4 Peruvian hospitals." International Society for Infectious Diseases (Int J Infect Dis), 2015. http://hdl.handle.net/10757/582607.
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Objectives
To report the incidence, epidemiology, and clinical features of Bordetella pertussis in Peruvian infants under 1 year old.
Patients and methods
A prospective cross-sectional study was conducted in five hospitals in Peru from January 2010 to July 2012. A total of 392 infants under 1 year old were admitted with a clinical diagnosis of whooping cough and tested for B. pertussis by PCR.
Results
The pertussis toxin and IS481 genes were detected in 39.54% (155/392) of the cases. Infants aged less than 3 months were the most affected, with a prevalence of 73.55% (114/155). The most common household contact was the mother, identified in 20% (31/155) of cases. Paroxysm of coughing (89.03%, 138/155), cyanosis (68.39%, 106/155), respiratory distress (67.09%, 104/155), and breastfeeding difficulties (39.35%, 61/155) were the most frequent symptoms reported.
Conclusion
An increase in pertussis cases has been reported in recent years in Peru, despite national immunization efforts. Surveillance with PCR for B. pertussis is essential, especially in infants less than 1 year old, in whom a higher rate of disease-related complications and higher mortality have been reported.This 312 work was supported by Sanofi Aventis del Peru. Conflict 313 of interest: On behalf of all authors, the corresponding author 314 states that there are no conflicts of interest or funding related 315 to this study
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del, Valle-Mendoza Juana, Wilmer Silva-Caso, Miguel Angel Aguilar-Luis, Valle-Vargas Cristina del, Erico Cieza-Mora, Johanna Martins-Luna, Ronald Aquino-Ortega, Andrea Silva-Vásquez, Jorge Bazán-Mayra, and Pablo Weilg. "Bordetella pertussis in children hospitalized with a respiratory infection: clinical characteristics and pathogen detection in household contacts." BioMed Central Ltd, 2018. http://hdl.handle.net/10757/624653.
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Objective: Describe the prevalence of Bordetella pertussis via PCR in children under 5 years old hospitalized as probable cases of pertussis and report the most common clinical features among them. Results: A positive PCR result for B. pertussis was observed in 20.5% of our samples (18/88), one-third of them were from infants between 2 and 3 months old. The most common symptoms were paroxysms of coughing (88.9%), difficulty breathing (72.2%), cyanosis (77.8%) and fever (50%). The mother was the most common symptomatic carrier (27.8%), followed by uncles/aunts (22.2%) among children with pertussis.This work was supported by fourth research incentive of the Universidad Peruana de Ciencias Aplicadas (UPC), Lima‑Peru.
Revisión por pares
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LaFever, Kristin E. "Spatial and temporal winter territory use and behavioral responses of whooping cranes to human activities." [College Station, Tex. : Texas A&M University, 2006. http://hdl.handle.net/1969.1/ETD-TAMU-1877.
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J, Lawrence Andrew. "Rapid laboratory diagnosis of Bordetella pertussis infection (whooping cough) using seriological and DNA based detection techniques /." Title page, contents and summary only, 1994. http://web4.library.adelaide.edu.au/theses/09S.M/09s.ml419.pdf.
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Zorzeto, Tatiane Queiroz. "Resposta humoral e celular de lactentes vacinados com pertussis celular total ou modificada pela extração de lipopolissacarideo." [s.n.], 2008. http://repositorio.unicamp.br/jspui/handle/REPOSIP/311081.
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Orientador: Maria Marluce dos Santos VilelaDissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciencias Medicas
Made available in DSpace on 2018-08-11T01:20:12Z (GMT). No. of bitstreams: 1 Zorzeto_TatianeQueiroz_M.pdf: 1695884 bytes, checksum: 267f0534bc256440762ad9d78bc6402d (MD5) Previous issue date: 2008
Resumo: A associação temporal de eventos adversos de variada gravidade à imunização com pertussis celular total (DTP) tem estimulado o desenvolvimento de vacinas antipertussis menos reatogênicas. Este ensaio clínico fase I visou à avaliação da imunogenicidade da vacina pertussis celular modificada pela extração do lipopolissacarídeo (DTPm) em comparação com a vacina convencional (DTP). Um total de 234 lactentes foi imunizado aos dois, quatro e seis meses de idade com DTPm ou DTP. Os títulos de anticorpos para os componentes pertussis, tétano, difteria e hepatite B foram determinados um mês após a terceira dose de vacina. A proliferação de células T CD3+ foi avaliada por citometria de fluxo após seis dias de cultivo de células mononucleares de sangue periférico estimuladas com células inativadas de B. pertussis ou com fitohemaglutinina (PHA). Células CD4+, CD8+ e TCR ?d+ foram identificadas no gate de blastos. Os níveis de IFN-?, TNF-a, IL-4 e IL-10 no sobrenadante de cultura foram quantificados por ensaio imunoenzimático (ELISA). A vacina modificada DTPm mostrou-se inferior à DTP quanto ao título de anticorpos antipertussis, mas não houve diferença de resposta aos outros componentes vacinais avaliados. A porcentagem líquida de blastos sob estímulo da B. pertussis foi menor no grupo que recebeu três doses de DTPm (mediana de 3,9% para DTPm e 6,2% para DTP, p=0,029), mas as freqüências de células CD4+, CD8+ e ?d+ em proliferação e as concentrações de citocinas não diferiram entre os grupos. A vacina DTPm não apresentou, portanto, imunogenicidade similar à da vacina DTP convencional nos ensaios laboratoriais
Abstract: Concerns about systemic reactions after immunization with whole-cell pertussis (wP) have stimulated efforts to produce less reactogenic vaccines. This phase I comparative trial aimed the efficacy evaluation of a cellular pertussis vaccine with low lipopolysaccharide (LPS) content (wPlow) in comparison to conventional wP vaccine. A total of 234 infants was vaccinated at 2, 4, and 6 months with conventional wP or wPlow. Serum antibody titers to pertussis, diphtheria, tetanus and hepatitis B were measured 1 month after the third dose of vaccine. Proliferation of CD3+ T cells was evaluated by flow cytometry after 6 days of peripheral blood mononuclear cells culture, with heat-killed B. pertussis or phytohemagglutinin (PHA) stimulation. CD4+, CD8+ and TCR ?d+ cells were identified in the gate of blast lymphocytes. IFN-?, TNF-a, IL-4 and IL-10 levels in supernatants were determined by ELISA. wPlow was inferior to wP in terms of anti-pertussis titers, but there was no diference in other serum antibody evaluations. Net percent blasts in cultures with B. pertussis was lower in the group vaccinated with wPlow (medians of 3.9% and for wPlow and 6.2% for wP; p=0.029), but the frequency of proliferating CD4+, CD8+ and ?d+ cells and the cytokine concentrations in supernatants were similar between vaccination groups. Therefore, wPlow wasn't as imunogenic as conventional wP in experimental evaluations
Mestrado
Saude da Criança e do Adolescente
Mestre em Saude da Criança e do Adolescente
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Laverty, Meghan. "Association Between Maternal Pertussis Vaccination During Pregnancy and Early Childhood Health Outcomes." Thesis, Université d'Ottawa / University of Ottawa, 2020. http://hdl.handle.net/10393/40084.
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Background: Little is known about whether there are any longer-term adverse health effects in children following Tdap (tetanus, diphtheria, and acellular pertussis) vaccination during pregnancy. Objective: To assess the association between maternal Tdap vaccination during pregnancy and risk of the following early childhood adverse health outcomes: (1) infections (upper and lower respiratory tract infections, gastrointestinal infections, and otitis media), (2) pediatric asthma, (3) neoplasm, (4) vision or hearing loss, and (5) urgent and in-patient health services utilization. Methods: This retrospective cohort study used multiple linked health administrative databases in the province of Ontario, Canada containing vaccine information in mothers and information on health outcomes in their children up to age 6 years. Infants exposed to prenatal Tdap were matched 1:5 with unexposed infants and outcomes were compared using hazard ratios and incidence rate ratios. Results: No significant adverse associations between prenatal Tdap and our study outcomes were observed. Inverse associations were found with upper respiratory infections (adjusted incidence rate ratio [aIRR]: 0.96, 95% CI: 0.93-0.99), lower respiratory infections (aIRR: 0.93, 95% CI: 0.89-0.98), gastrointestinal infections (aIRR: 0.88, 95% CI: 0.82-0.94), and urgent and in-patient health service utilization (aIRR: 0.95, 95% CI: 0.94-0.97). Conclusions: Our findings support the long-term safety of Tdap administration in pregnancy.
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Del, Valle Mendoza Juana Mercedes, Oré Veronica Casabona, Helasvuo Veronica Petrozzi, Tapia Angela Cornejo, Pablo Weilg, Maria J. Pons, Mora Erico Cieza, Mayra Jorge Bazán, Pacherres Hernan Cornejo, and Joaquin Ruiz. "Bordetella pertussis diagnosis in children under five years of age in the Regional Hospital of Cajamarca, Northern Peru." The Journal of Infection in Developing Countries (JIDC), 2015. http://hdl.handle.net/10757/605267.
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Introduction: Bordetella pertussis is an important human pathogen that causes whooping cough (pertussis), an endemic illness responsible of
significant morbidity and mortality, especially in infants and children. Worldwide, there are an estimated of 16 million cases of pertussis,
resulting in about 195,000 child deaths per year. In Peru, pertussis is a major health problem that has been on the increase despite
immunization efforts. The objective of this study was to determine the prevalence of B. pertussis among children under five years of age
suspected to have whopping cough in Cajamarca, Peru.
Methodology: Children diagnosed with whooping cough admitted to the Hospital Regional de Cajamarca from August 2010 to July 2013
were included. Nasopharyngeal samples were obtained for B. pertussis culture and polymerase chain reaction (PCR) detection.
Results: In 133 children, the pertussis toxin and IS481 gene were detected in 38.35% (51/133) of the cases by PCR, while only 9.02%
(12/133) of the Bordetella cultures were positive. The most frequent symptoms in patients with positive B. pertussis were paroxysm of
coughing 68.63% (35/51), cyanosis 56.86% (29/51), respiratory distress 43.14% (22/51), and fever 39.22% (20/51). Pneumonia and acute
bronchial obstructive syndrome were present in 17.65% (9/51) and 13.72% (7/51) of the cases, respectively.
Conclusions: B. pertussis is responsible for an important proportion of whooping cough in hospitalized children in Cajamarca. Epidemiologic
surveillance programs for B. pertussis are essential in Peru, especially in children who could most benefit from the vaccine.
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Black, Andrew James. "The stochastic dynamics of epidemic models." Thesis, University of Manchester, 2010. https://www.research.manchester.ac.uk/portal/en/theses/the-stochastic-dynamics-of-epidemic-models(196cf4a1-2db2-4696-bc64-64fb28cb0b7d).html.
Full textAbstract:
This thesis is concerned with quantifying the dynamical role of stochasticity in models of recurrent epidemics. Although the simulation of stochastic models can accurately capture the qualitative epidemic patterns of childhood diseases, there is still considerable discussion concerning the basic mechanisms generating these patterns. The novel aspect of this thesis is the use of analytic methods to quantify the results from simulations. All the models are formulated as continuous time Markov processes, the temporal evolutions of which is described by a master equation. This is expanded in the inverse system size, which decomposes the full stochastic dynamics into a macroscopic part, described by deterministic equations, plus a stochastic fluctuating part. The first part examines the inclusion of non-exponential latent and infectious periods into the the standard susceptible-infectious-recovered model. The method of stages is used to formulate the problem as a Markov process and thus derive a power spectrum for the stochastic oscillations. This model is used to understand the dynamics of whooping cough, which we show to be the mixture of an annual limit cycle plus resonant stochastic oscillations. This limit cycle is generated by the time-dependent external forcing, but we show that the spectrum is close to that predicted by the unforced model. It is demonstrated that adding distributed infectious periods only changes the frequency and amplitude of the stochastic oscillations---the basic mechanisms remain the same. In the final part of this thesis, the effect of seasonal forcing is studied with an analysis of the full time-dependent master equation. The comprehensive nature of this approach allows us to give a coherent picture of the dynamics which unifies past work, but which also provides a systematic method for predicting the periods of oscillations seen in measles epidemics. In the pre-vaccination regime the dynamics are dominated by a period doubling bifurcation, which leads to large biennial oscillations in the deterministic dynamics. Vaccination is shown to move the system away from the biennial limit cycle and into a region where there is an annual limit cycle and stochastic oscillations, similar to whooping cough. Finite size effects are investigated and found to be of considerable importance for measles dynamics, especially in the biennial regime.
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Williams, Emily G. "Whooping cough among Western Cree and Ojibwa fur-trading communities in subarctic Canada : a mathematical-modeling approach /." free to MU campus, to others for purchase, 2004. http://wwwlib.umi.com/cr/mo/fullcit?p1421166.
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Del, Valle-Mendoza Juana, Valle-Vargas Cristina del, Ronald Aquino-Ortega, Valle Luis J. Del, Erico Cieza-Mora, Wilmer Silva-Caso, Jorge Bazán-Mayra, et al. "Clinical characteristics and molecular detection of in hospitalized children with a clinical diagnosis of whooping cough in Peru." Tehran University of Medical Sciences, 2021. http://hdl.handle.net/10757/655868.
Full textAbstract:
Pertussis is an infectious disease caused by the Gram-negative bacterium Bordetella pertussis. In Peru, actual public health programs indicate that vaccination against B. pertussis must be mandatory and generalized, besides all detected cases must be reported. The objective of this study was to determine the prevalence of B. pertussis among children under five years of age with a presumptive diagnosis of whopping cough in Cajamarca, a region located in northern Peru.Background and Objectives: Pertussis is an infectious disease caused by the Gram-negative bacterium Bordetella pertussis. In Peru, actual public health programs indicate that vaccination against B. pertussis must be mandatory and generalized, be-sides all detected cases must be reported. The objective of this study was to determine the prevalence of B. pertussis among children under five years of age with a presumptive diagnosis of whopping cough in Cajamarca, a region located in northern Peru. Materials and Methods: The population of this cross-sectional study were children under 5 years old hospitalized as presumptive cases of pertussis during December 2017 to December 2018. The nasopharyngeal samples were analyzed by real-time PCR for the detection of B. pertussis. Results: B. pertussis was identified as PCR + in 42.3% of our sample (33/78). The clinical presentation that was observed most frequently includes paroxysmal coughing (97%), difficulty breathing (69.7%), cyanosis (72.7%) and post-tussive em-esis (60.6%). Additionally, pneumonia was the most observed complication (33.3%). Four of the patients with PCR+ for B. pertussis presented only lymphocytosis, five only leukocytosis, two patients with decreased leukocytosis and lymphocytes and only one patient with leukopenia and relative lymphocytosis. There was a percentage of 84.8% of unvaccinated children in the PCR+ group. Finally, the mother was the most frequent symptom carrier (18.2%). Conclusion: In conclusion, in the studied population there is a high rate of PCR+ cases for B. pertussis. Laboratory values may show leukopenia or lymphopenia in patients with pertussis. It is necessary to use appropriate laboratory diagnostic tests in all infants with respiratory symptoms for B. pertussis. Since, the clinical diagnosis overestimates the diagnosis of pertussis.
Revisión por pares
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Warren, Andrew Eugene. "The use of inhaled beclomethasone to decrease the duration of paroxysmal coughing in pediatric patients with pertussis : results and methodologic issues in a randomized clinical trial /." St. John's, NF : [s.n.], 1997.
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Hegerle, Nicolas. "Evolution of Bordetella pertussis and Bordetella parapertussis under acellular Pertussis vaccine pressure : What future for whooping cough and Pertussis vaccination ?" Paris 7, 2014. http://www.theses.fr/2014PA077038.
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La coqueluche est une maladie respiratoire dangereuse pour les nouveaux nés non vaccinés. Celle-ci est causée par Bordetella pertussis et Bordetella parapertussis, deux bactéries à gram négatif dont le seul hôte connu est l'homme. L'introduction de la vaccination contre B. Pertussis, le pathogène majoritairement responsable de cette maladie, à grandement impacté l'épidémiologie de la coqueluche ainsi que les populations de B. Pertussis circulant dans la population humaine. Les premier vaccins introduit dans les 1950 et utilisés pour la primo vaccination et le rappel enfant, dit à germes entier, ont permit de contrôler les isolats circulants semblables aux souches vaccinales. L'introduction des vaccines acellulaire, ne ciblant que quelques antigènes bactériens, pour le rappel adolescent puis pour la primo-vaccination et le rappel adulte a changé la donne. En plus d'un changement de l'immunité induite par le vaccin on a assisté à une augmentation de l'immunité vaccinale au sein de la population. Quelques années après l'introduction des vaccins acellulaires nous avons mis en évidence l'augmentation de la prévalence d'isolat n'exprimant plus un des antigènes vaccinaux alors que la population restait inchangée par rapport à l'ère post-germe entier au niveau génétique. La caractérisation phénotypique des ces isolats nous a permit de montrer qu'ils n'étaient pas plus virulent dans différents modèles in vitro ou in vivo mais qu'ils pouvaient présenter un avantage sélectif dans population immunisée par des vaccins acellulaires ciblant cet antigène en question. Bien que les vaccins restent efficaces, la surveillance doit continuerWhooping cough is an acute respiratory disease life threatening for unvaccinated young children. It is caused by Bordetella pertussis and Bordetella parapertussis, two gram-negative bacteria restricted to humans. The introduction of vaccination against B. Pertussis in the 1950s greatly changed the epidemiology of pertussis as well as the bacterial population itself. Whole cell vaccines were first introduced for children immunization and first booster and enabled to control isolates similar to strains included in the vaccines. Acellular pertussis vaccines, only targeting few bacterial antigens, were later introduced for adolescent booster vaccination before being generalized to the whole population, including adults and new-borns. In addition to a change in the type of vaccine-induced immunity, B. Pertussis also had to face increased herd immunity. Few years alter acellular pertussis vaccine introduction we demonstrated a temporal increase in the prevalence of isolates lacking the production of one vaccine antigen, pertactin, while the population remained quite monomorphic at the genetic level as compare to the post-whole cell vaccine era (allelic stability of known virulence factors). We characterized these isolates at the phenotypic level and demonstrated that they remain as virulent in different in vitro and in vivo models of host pathogen interaction while they might present a selective advantage in an acellular immunized background targeting pertactin. Although vaccines remain effective, surveillance must continue to follow the evolution of these isolate prevalence and the possible impact of pertactin loss on vaccine effectiveness
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McMillen, Janet L. "Productivity and movements of the greater sandhill crane population at Seney National Wildlife Refuge: potential for an antroduction of whooping cranes." The Ohio State University, 1988. http://rave.ohiolink.edu/etdc/view?acc_num=osu1298917399.
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McMillen, Janet L. "Productivity and movements of the greater sandhill crane population at Seney National Wildlife Refuge : potential for an introduction of whooping cranes /." The Ohio State University, 1988. http://rave.ohiolink.edu/etdc/view?acc_num=osu1487588939090135.
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Correia, Carolina Argondizo. "Influência da vacinação com dTpa em gestantes no perfil da resposta imunológica contra a Bordetella pertussis na criança." Universidade de São Paulo, 2018. http://www.teses.usp.br/teses/disponiveis/99/99131/tde-18052018-101733/.
Full textAbstract:
A Bordetella pertussis é a bactéria causadora da coqueluche, doença infectocontagiosa que tem reemergido em diversos países apesar das altas coberturas vacinais. Como os indivíduos mais afetados são crianças menores de seis meses de idade, a vacinação de gestantes com uma dose de vacina adsorvida contra tétano, difteria e coqueluche (B. pertussis acelular - dTpa) foi proposta. O objetivo é promover a transferência de altos títulos de anticorpos maternos ao feto e recémnascido, resultando na sua proteção até que o seu esquema vacinal esteja concluído. Esta estratégia é empregada em diversos países e no final de 2014 foi introduzida no Brasil. Entretanto, pouco se sabe sobre sua eficácia, efetividade e interação com a posterior vacinação da criança, que é realizada com a formulação de células inteiras (DTP), enquanto outros países substituíram-na pela formulação acelular (DTPa). Sabe-se que a vacinação materna induz anticorpos que ajudam a proteger recém-nascidos da infecção, mas podem, também, neutralizar a vacinação da criança, diminuindo a eficiência vacinal, além de promover a transferência de antígenos para o feto, induzindo desvio do perfil de resposta. Esses aspectos são particularmente importantes e devem ser investigados quando novos programas de vacinação materna são implantados. Sendo assim, o objetivo do presente estudo foi avaliar a resposta celular contra B. pertussis em parturientes e neonatos no dia do parto, e em lactentes após vacinação completa contra a coqueluche com DTP no primeiro ano de vida, comparando-se o grupo proveniente de mães que foram vacinadas durante a gestação com as que não receberam a dose de reforço. As células mononucleares do sangue periférico e de cordão umbilical dos participantes foram isoladas e estimuladas com antígeno bacteriano ou antígeno policlonal em cultura. Após o tempo estipulado, verificou-se o perfil de expressão gênica por qPCR nas células e a concentração de citocinas de interesse em sobrenadante, por meio de citometria de fluxo. Os dados obtidos mostram que a vacina na gestação induz resposta celular favorável à proteção contra infecção em gestantes, com produção de citocinas de perfil Th1 após estímulo antigênico, enquanto que nos neonatos poucas citocinas estavam acima dos limites de detecção. Quando se comparou a resposta de lactentes nascidos de mães vacinadas ou não durante a gestação, não foi encontrada diferença significativa nos níveis de citocinas produzidos, sugerindo que os anticorpos maternos presentes durante o desenvolvimento fetal e os primeiros meses de vida das crianças não interferiu no processo de reconhecimento e geração de resposta específica. Apesar do número amostral limitado, este trabalho mostra um panorama da interação da vacinação materna com a resposta celular à vacinação da criança no seu primeiro ano de vida, mostrando que a princípio a dose de reforço com dTpa durante a gestação é capaz de gerar resposta celular protetora nas gestantes, prevenindo-as de transmitir a doença a seus filhos, e a resposta humoral produzida não interfere na geração de resposta celular de seus filhos após o seu próprio esquema de imunização.Bordetella pertussis is the bacterial agent of whooping cough, an infectious and contagious re-emerging disease despite high vaccine coverage. As the most affected are children younger than six months of age, a dose of tetanus/diphtheria/pertussis vaccine (acellular pertussis - Tdap) in pregnancy was proposed. It is aimed to promote the transfer of high maternal antibodies\' titres to the foetus and newborns, resulting in their protection until their own vaccination scheme is completed. This strategy is present in several countries, and it was implemented in Brazil by the end of 2014. However, little is known about its efficacy, effectiveness and interaction with the subsequent child\'s vaccination, which comprehends the whole-cell formulation (DTP), while other countries replaced it for the acellular version (DTaP). Maternal vaccination induces antibodies that would protect newborns from infection, but they can also neutralize the effect of the child vaccination, reducing the vaccine efficiency. Besides, it can promote the transfer of antigens to the foetus, which can induce deviation in the response pattern. These aspects are particularly important and should be investigated when new maternal vaccination programs are implemented. Therefore, the aim of this project was to evaluate the cellular response against B. pertussis in women at labour and their neonates, as in young children after complete pertussis vaccination by the first year of life, comparing groups from vaccinated and non-vaccinated women during pregnancy. Peripheral blood and cord blood mononuclear cells were isolated and stimulated with bacterial or polyclonal antigen in culture. After the stipulated time the gene expression profile and cytokine concentration were evaluated by qPCR and flow cytometry. Data from the present study show that pregnancy vaccination inducing favourable cellular response for protection against infection in pregnant women, with Th1 cytokines\' production upon antigenic stimulus, while in newborns few cytokines were detected above the detection limit. When comparing children\'s response, born either from vaccinated or unvaccinated mothers, cytokine levels were not significantly different, suggesting that maternal antibodies present during foetal development and first months of life do not interfere with recognition and cellular response generation to vaccination. Despite limited sample size, this work shows a broader view of interaction between maternal and child vaccination, showing that the Tdap boost dose during pregnancy may be able to generate protective response in women, preventing them to transmit the disease to children, and the induced humoral response may not interfere in generating cellular response in children after their own vaccination.
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Yilmaz, Cigdem. "Immune Responses Against The Recombinant Fimx And Putative Peptidyl-prolyl Cis-trans Isomerase From Bordetella Pertussis." Master's thesis, METU, 2011. http://etd.lib.metu.edu.tr/upload/12613684/index.pdf.
Full textAbstract:
Whooping cough (pertussis) is a highly contagious respiratory infection caused by Bordetella pertussis. It becomes widespread among adolescent and adults as well as infants. Although availability of effective pertussis vaccines seems to decrease the incidence of the disease, B. pertussis circulation in population has not been eliminated. It is thought that the antigenic drifts in major protective antigens and continued circulation of B. pertussis strains will result in gradual loss of the efficacy of the current pertussis vaccines. Therefore, development of more effective acellular pertussis vaccines with conserved protective proteins is a convenient strategy to provide a better protection against whooping cough.
In this study, immune responses against putative peptidyl-prolyl cis-trans isomerase (PPIase) which was shown to be immunogenic in B. pertussis for the first time by our immunoproteome group and FimX whose expression was found higher in our local Saadet strain were determined in mice. The genes encoding FimX and putative PPIase were amplified by PCR, cloned into pGEM®-T Easy vector and sequenced. The genes were then introduced into pET-28a (+) vector and they were expressed in Escherichia coli BL21(DE3) cells. The recombinant proteins were purified by His-tag affinity chromatography and dialyzed. After Western blot analyses, 20 µ
g and 80 µ
g recombinant FimX and 80 µ
g recombinant putative PPIase were used to immunize BALB/c mice (16-18 g) at day 0 and 21. The mice were challenged intranasally with 2.5 x 109 live B. pertussis Saadet cells. Before second immunization and challenge, the sera were collected to carry out ELISA for measurement of serum-specific IgG levels. According to ELISA results, IgG levels in the mice immunized with 20 µ
g and 80 µ
g recombinant FimX were found significantly higher than in control groups at both first and second vaccinations (p<
0.01). On the other hand, immunization with 160 µ
g recombinant putative PPIase provided a significant increase in IgG level (p<
0.05) only at second vaccination. The lungs of the mice were removed at day 2, 5, 8 after challenge and bacterial colonization was determined. No significant decrease in bacterial colonization was observed in the lungs of the mice immunized with 20 µ
g and 80 µ
g recombinant FimX and 80 µ
g recombinant putative PPIase with respect to control groups. After respiratory challenge and second immunization (at day 30) with 20 µ
g and 80 µ
g recombinant FimX, the spleens of the mice were removed and a spleen cell culture was obtained. Supernatants were collected after induction of the cells with the recombinant protein and cytokine ELISA was carried out to measure IFN-&gamma
level. No significant difference was observed between control and vaccinated mice in terms of IFN-&gamma
production.
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Dupré, Elian. "La régulation de la virulence chez Bordetella pertussis : BvgS, modèle original de capteur de système à deux composants." Thesis, Lille 2, 2013. http://www.theses.fr/2013LIL2S022/document.
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La virulence de Bordetella pertussis, agent de la coqueluche, est liée à un arsenal de facteurs de virulence dont l’expression est régulée par le système à deux composants BvgAS. BvgA est le régulateur de réponse et BvgS le capteur du système, qui possède 3 domaines putatifs de perception de signaux. Il s’agit de 2 domaines périplasmiques « Venus FlyTrap » (VFT), reliés par un segment transmembranaire à un domaine PAS (Per-ARNT-Sim) cytoplasmique qui fait la jonction avec l’histidine-kinase. Les signaux perçus par ces domaines capteurs sont inconnus, mais une température de 37°C est suffisante pour maintenir le système actif en laboratoire. Cette activité peut être modulée négativement par des composés chimiques, comme le MgSO4 ou le nicotinate, qui à concentrations suffisantes entraînent le passage de la bactérie en phase avirulente.Nous nous sommes intéressés aux domaines VFT de BvgS. Ces domaines, ubiquitaires, sont composés de 2 lobes reliés par une charnière délimitant une cavité qui permet la fixation d’un ligand spécifique stabilisant le VFT sous une forme fermée.Les domaines VFT de BvgS ont pu être cristallisés et s’organisent en un dimère entrelacé définissant de larges interfaces entre les 4 VFTs. Les VFT2 sont fermés sans ligand et les VFT1 ouverts, et la fermeture artificielle de ces domaines par des ponts disulfure a montré qu’il s’agit de la conformation active de BvgS. L’importance des interfaces entre les domaines VFT pour la fonction de BvgS a été démontrée par mutagenèse dirigée. Un signal positif proviendrait du périplasme pour être transmis à travers la membrane par les interfaces entre les VFT et intégré via un couplage fonctionnel en trans entre ces domaines et les hélices pré-membranaires, dites H19.Ces hélices se prolongeraient à travers la membrane et dans le cytoplasme jusqu’au domaine PAS. Les domaines PAS sont ubiquitaires, avec une structure fortement conservée en feuillet à 5 brins recouvert d’hélices délimitant une cavité. Ils sont impliqués dans diverses fonctions biologiques, selon leur capacité de liaison d’un ligand. Certains domaines PAS fonctionneraient sans ligand et pourraient servir d’adaptateurs ou d’amplificateurs de signal.Nous avons pu mettre en évidence la capacité de dimérisation de PASBvg, confirmant la nature dimérique du capteur BvgS. Des substitutions de résidus de la cavité de PASBvg indiqueraient que l’intégrité de la cavité de PASBvg est nécessaire au passage de signaux positifs et négatifs provenant du périplasme. La fixation de ligand dans la cavité n’a pu être démontrée mais n’est pas exclue. D’autre part, certains résidus sont nécessaires au couplage du domaine PAS avec ses hélices flanquantes pour la transmission de signal. La perte de ces interactions déstabilise significativement PASBvg et rend BvgS inactif.Un message positif proviendrait du périplasme et serait maintenu par le domaine PAS, dans une conformation rigide, permettant aussi la transmission des signaux modulateursVirulence of Bordetella pertussis, the whooping cough agent, is due to a plethora of virulence factors which expression is regulated by the two-component system BvgAS. BvgA is a classical response regulator and BvgS the sensor. BvgS contains 3 putative sensor domains, 2 periplasmic Venus FlyTrap (VFT), linked through a transmembrane segment to a cytoplasmic PAS domain preceding the histidine-kinase. Signals perceived by those sensor domains are still unknown, but a 37°C temperature is sufficient to maintain the system active under laboratory conditions. This activity can be down-modulated by chemical compounds, such as MgSO4 or nicotinate, which at sufficient concentration allows the bacteria to switch to avirulent phase.We investigated the role of BvgS VFT domains. VFTs are ubiquitous domains composed of 2 lobes linked by a hinge hence forming a cleft where a specific ligand can bind and stabilize the VFT in its closed conformation.BvgS VFT domains were crystalized and form an intricate dimer defining large interfaces between the 4 VFTs. VFT2s are closed without a ligand and VFT1s are opened, artificial closure of these domains via a disulfide bond indicates that this is the active conformation of BvgS. The role of the interfaces was probed by site-directed mutagenesis. A positive signal might originate from the periplasm to be transmitted through the membrane by the interfaces and integrated by a functional coupling between the VFT2s and the helices preceding the membrane, H19.These helices should be continued through the membrane and the cytoplasm to the PAS domain. Pas domains are ubiquitous with a highly conserved structure, a 5 stranded sheet surrounded by helices defining a cavity. Pas domains are involved in a wide variety of physiological processes, depending on their ability to bind a ligand. Some PAS might function without a ligand and could then be signal adaptors or amplifiers.We demonstrated PASBvg was dimeric, confirming the dimeric nature of BvgS. Cavity residues were substituted indicating that integrity of the cavity is necessary to maintain activity and modulation capacity coming from the periplasmic moiety. Ligand binding wasn’t demonstrated but couldn’t be excluded. Some residues are needed for the correct coupling of the PAS domain to its flanking helices and hence signal transmission. Loss of these connections generates a strong destabilization of PASBvg and turns BvgS inactive.A positive signal might come from the periplasmic moiety and shoul be maintaines by the PAS domain, which is in a rigid conformation also allowing the transmission of negative signals
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Wang, Kay Yee. "Mycoplasma pneumoniae and Bordetella pertussis in patients with persistent cough in primary care." Thesis, University of Oxford, 2012. http://ora.ox.ac.uk/objects/uuid:74b15a97-708d-4927-b0aa-ce802f4af2aa.
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Background: Persistent cough following an acute respiratory tract infection is a challenging and frequently encountered problem in primary care. Mycoplasma pneumoniae (M. pneumoniae) and Bordetella pertussis (pertussis) particularly predispose patients to persistent cough. Whilst the incidence of M. pneumoniae is highest in children, pertussis may also occur in adults. Method: Four studies were conducted for this thesis. First, a systematic review to assess the diagnostic accuracy of symptoms and signs in the clinical recognition of M. pneumoniae. Second, a retrospective analysis of a cohort of children with persistent cough to assess the prognostic value of diagnosing M. pneumoniae. Third, a prospective cohort study to estimate the prevalence of M. pneumoniae and pertussis in children with persistent cough following recent changes in vaccination policy. Fourth, a double-blind randomised placebo-controlled trial to determine the effectiveness of montelukast in the treatment of persistent cough and pertussis-induced cough in adults. Results: M. pneumoniae and pertussis can each be found in one-sixth of children who present in primary care with persistent cough. Although coverage with the preschool pertussis booster vaccine is high, its efficacy wanes rapidly, with the likelihood of pertussis increasing by 30% per year after vaccination. Montelukast is not an effective treatment for persistent cough, but may be an effective treatment for pertussis-induced cough. Median duration of cough in children with M. pneumoniae is only one-third of that in children with pertussis (39 days versus 118 days). However, the diagnostic accuracy of symptoms and signs in the clinical recognition of M. pneumoniae is limited. Since M. pneumoniae occurs in cyclical epidemics, clinicians should consider current prevalence of M. pneumoniae when making a clinical diagnosis. Conclusions: Diagnosing M. pneumoniae and pertussis can help clinicians give patients an explanation for their cough and inform them about its likely prognosis. At the moment, clinicians should adopt a conservative approach to managing postinfectious persistent cough. A further trial is needed to assess the efficacy of montelukast for the treatment of pertussis-induced cough.
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Galhardo, Cynthia Soares. "Fatores que determinam a produção de IL-12 em macrófagos murinos ativados por Bordetella pertussis e B. parapertussis." Universidade de São Paulo, 2013. http://www.teses.usp.br/teses/disponiveis/87/87131/tde-15052014-101030/.
Full textAbstract:
Bordetella pertussis e B. parapertussis são agentes etiológicos da coqueluche. A IL-12 liga a imunidade inata e adaptativa. Investigamos alguns mecanismos que controlam a síntese de IL-12 em macrófagos medulares murinos (MfDM) ativados in vitro com estas duas espécies de bactérias. Demonstramos que IL-12p40 e TNF-a foram produzidos pelos MfDM ativados com qualquer uma das bactérias. A síntese de IL-12p40 foi dependente de TNF-a, MyD88 e NFkB e independente de MAPK p38 e ERK 1/2. Durante a estimulação com B. pertussis a produção de IL-12p40 foi dependente de TLR-4, mas com B. parapertussis envolveu outras vias independentes de MyD88 e TLR-4. Estas bactérias não induziram a síntese de IL-12p70, necessitando de sinais moleculares adicionais de IFN-g, que aumentou a síntese desta citocina. A produção de IL-12 p70 aumentou após o bloqueio das vias PI3K, MAPK p38 e ERK1/2 assim como após a adição exógena de PT sobre MfDM ativados com B. parapertussis. Portanto, diversas vias de sinais dependentes e independentes de TLR-4 controlam a produção de IL-12 neste modelo.Bordetella pertussis and B. parapertussis are etiological agents of whooping cough. IL-12 links the innate and adaptive immunity. We investigated the ability of both bacteria to modulate IL-12 by in vitro activation of bone marrow derived macrophages (MfDM). We demonstrated that IL-12p40 and TNF-a were produced after stimulation of cells with either bacterium. IL-12p40 production was dependent on TNF-a, MyD88 and NFkB but independent of MAPK p38 and ERK 1/2. During B. pertussis activation the production of IL-12p40 was dependent on TLR-4, while B. parapertussis activation was MyD88 and TLR-4 independent. However, the bacteria alone did not induce IL-12p70 synthesis, requiring IFN-g as an additional signal. Evidences indicated MAPK p38, ERK1/2 and PI3K during B. pertussis and B. parapertussis activation, as well as the exogenous addition of PT to B. parapertussis activated MfDM, was critical for the up regulation of IL-12p70. This finding indicates that different TLR-4 dependent and independent signaling pathways may control the production of IL-12 in this model.
46
Freitas, Angela Carvalho. "Avaliação de novas estratégias vacinais contra a coqueluche no município de São Paulo." Universidade de São Paulo, 2008. http://www.teses.usp.br/teses/disponiveis/6/6132/tde-26082008-153502/.
Full textAbstract:
Introdução: A coqueluche é caracterizada por tosse paroxística, pode levar menores de um ano de idade ao óbito, deixar seqüelas e exacerbar quadros respiratórios crônicos. A imunidade após a doença ou vacina não é para toda a vida. Nos países desenvolvidos, apesar de altas coberturas vacinais e do controle da doença entre as décadas de 50 e 80, desde o final dos anos 80 é observado o aumento dos casos em adolescentes, adultos e lactentes, sendo indicado o reforço vacinal para adolescentes e adultos. No Brasil a doença aparenta estar sob controle, mas há um estudo teórico que demonstra a possibilidade de aumento dos casos. Objetivo: Avaliar novas estratégias de reforço vacinal contra a coqueluche no município de São Paulo. Metodologia: Desenvolvimento de um modelo matemático determinístico, dinâmico e dependente da idade dos indivíduos. Simulações com o esquema vacinal atual e: (i) novo reforço aos 12 anos com coberturas vacinais de 10%, 35%, 45% e 70%; (ii) reforços aos 12 anos e aos 20 anos de idade, com 35% e 70% de cobertura, respectivamente. Introdução de contato heterogêneo da população com o uso de uma matriz de contato. Fontes dos dados: Banco de dados do Sistema Único de Saúde (DATASUS), Centro de Vigilância Epidemiológica da Secretaria de Estado da Saúde de São Paulo e a literatura nacional e internacional. Uso dos programas Berkeley Madonna® para resolução das equações diferenciais e Microsoft Excel® para o cálculo da matriz de contato e das forças de infecção. Realização de teste de sensibilidade do modelo. Resultados: A vacinação com cobertura de 10% aos 12 anos de idade reduziu os casos entre os próprios adolescentes (10 a 19 anos); com cobertura de 35%, 45% e 70% reduziu os casos em 59%, 65% e 73%, respectivamente; a vacinação em conjunto aos 12 anos com cobertura de 35% e aos 20 anos com cobertura de 70% reduziu 62% dos casos. Conclusões: Há benefício ao vacinar os adolescentes, inclusive com baixa cobertura vacinal, portanto tal estratégia demonstra-se promissora para o controle da coqueluche. Não houve ganho ao acrescentar apenas um reforço para os adultos (20 anos). Os resultados são concordantes com o que há na literatura e permitiram um primeiro panorama para auxiliar na abordagem do problema. Estudos com diferentes estratégias de vacinação de adultos e estudos de custo-benefício são recomendados.Background: Whooping cough is a respiratory tract infection caused by Bordetella pertussis and characterized by paroxysmal cough that usually causes complications for infants, including death, and for people with chronic respiratory diseases. Immunity against pertussis after infection or vaccination is not everlasting. Despite of high childhood immunization coverage and the disease control from the 50's to 80's, since late 80's developed countries notified high levels of pertussis in adolescents and adults. This reappearance has not being detected in Brazil yet, but at least one formal study has demonstrated the possibility of this change in the next years. Objective: Evaluating new pertussis vaccine's booster for adolescents and adults in São Paulo city. Methods: Development of a deterministic, compartmental and age-dependent model accounting for immunity waning. The data was retrieved from literature, Surveillance Center of the State of São Paulo (CVE), and the Brazilian national health data system (DATASUS). Data manipulation used Berkeley Madonna® and Microsoft Excel®. Vaccination strategies included the current vaccination scheme, plus (i) 10%, 35%, 45% or 70% vaccine coverage for those at the age of 12 and (ii) both 35% and 70% vaccine coverage at the ages 12 and 20, respectively. The Who Acquire Infection from Whom (WAIFW) matrices' method was used to assume age related transmission rates. Sensitivity analysis was performed. Results: Booster vaccination for 12 years youths, at 10% coverage, yields disease reduction only among adolescents (10 to 19 years); coverage up to 35% yields disease reduction for all ages; at 35%, 45% and 70% coverage, the reduction achieves 59%, 65% and 73%. Booster vaccination at 12 and 20 years, with coverage at 35% and 70% respectively, yields 62% cases reduction. Discussion: Results suggest that adolescent's vaccine booster could reduce pertussis occurrence for all ages, including infants, as also demonstrated by other authors. In contrast, only one vaccine booster for adults (20 years) achieves insignificant results. In conclusion, we have been able to demonstrate that, in São Paulo, the adolescent vaccine booster strategy is a promising police to further reduce whooping cough occurrence. However, cost effective analysis and other adults' vaccination strategies are recommended.
47
Prestes, Ana Fabíola Rollo de Oliveira. "Avaliação de adjuvantes em novas formulações de vacina tríplice bacteriana." Universidade de São Paulo, 2009. http://www.teses.usp.br/teses/disponiveis/87/87131/tde-28042009-111641/.
Full textAbstract:
As vacinas pertussis celulares apresentam certa reatogenicidade e as acelulares, menos reatogênicas, têm seu uso restrito, devido a seu alto custo. O Instituto Butantan desenvolveu uma vacina pertussis celular (Plow), com baixo teor de lipopolissacarídeo e outra acelular (Pa), por metodologia simples e econômica. Essas preparações, combinadas aos toxóides diftérico e tetânico (DTPlow e DTPa, respectivamente), foram comparadas à DTP tradicional, com diferentes adjuvantes: vitamina A, surfactante pulmonar, BCG, monofosforil lipídeo A (MPL) e Al(OH)3. A resposta humoral em camundongos foi semelhante para as diferentes formulações e independente do adjuvante utilizado. As vacinas induziram níveis equilibrados de IgG1/IgG2a anti-pertussis e a DTPlow mostrou-se menos reatogênica, induzindo níveis significativamente menores de IL-6 sérica. A adição de MPL sugeriu tendência de proteção contra a colonização nasotraqueal no grupo imunizado com DTPa e levou à indução de IFN-g nos grupos imunizados com DTP e DTPa, sugerindo possível atividade imunomodulatória para Th1.The whole cell pertussis vaccines present some reactogenicity and the acellular, less reactogenic, have prohibitive use due to its high cost. Instituto Butantan developed a less reactogenic whole cell pertussis vaccine (Plow), with low lipopolysaccharide content and an acellular vaccine (Pa), by simple and economic methodology. These preparations, combined to diphtheria and tetanus toxoids (DTPlow and DTPa, respectively), were compared with the traditional DTP, with different adjuvants: vitamin A, pulmonary surfactant, BCG, monophosphoryl lipid A (MPL) and Al(OH)3. The humoral immune response induced in mice by the different vaccine formulations, was similar and independent of the adjuvant used. The vaccines induced balanced levels of IgG1/IgG2a anti-pertussis and DTPlow showed to be less rectogenic, inducing lower levels of serum IL-6. The use of MPL suggested to induce higher protection against nasotracheal colonization in DTPa group and induced IFN-g in the DTP and DTPa groups, suggesting a possible immunemodulatory activity for Th1.
48
Ferronato, Angela Esposito. "Identificação de vírus respiratórios em lactentes internados com suspeita clínica de coqueluche." Universidade de São Paulo, 2017. http://www.teses.usp.br/teses/disponiveis/5/5141/tde-28032018-101026/.
Full textAbstract:
Introdução: a coqueluche é uma doença causada pela Bordetella pertussis (BP), sendo mais frequente e grave em lactentes menores de um ano de idade. Com a introdução da vacina, houve redução na incidência mundial da doença, porém nos últimos 10 anos observa-se uma recrudescência. Pode apresentar-se de forma menos característica em lactentes, especialmente antes do final do esquema vacinal para o primeiro ano de vida. O quadro clínico, nesses pacientes, pode ser semelhante ao das infecções por vírus respiratórios (VR) que são os agentes etiológicos mais frequentes nas infecções de vias aéreas, nessa faixa etária. São necessários estudos que avaliem a importância da pesquisa de VR em lactentes com suspeita clínica de coqueluche. Objetivos: em lactentes com suspeita de coqueluche: identificar as prevalências de BP, VR e codetecções; analisar e comparar as características clínicas e a evolução, segundo a etiologia identificada e analisar o impacto do diagnóstico etiológico sobre o uso de macrolídeos. Métodos: estudo de coorte prospectivo, com crianças menores de um ano de idade, hospitalizadas com suspeita clínica de coqueluche entre junho de 2014 e junho de 2016 e submetidas à pesquisa etiológica para identificação de BP (\"swab\" de nasofaringe para cultura e/ou PCR) e pesquisa de VR (aspirado de nasofaringe para imunofluorescência indireta). Dados clínicos, demográficos e evolutivos foram coletados com o preenchimento de protocolo clínico-laboratorial padronizado. Resultados: no período de estudo foram analisados 59 lactentes. Em 18 (30,5%) houve identificação de BP, em 23 (39%) de algum vírus respiratório. Em quatro (7%), houve codetecção de BP e algum VR. O vírus mais frequentemente identificado foi o VSR (73%). As características com maior sensibilidade para o diagnóstico de infecção por BP foram tosse seguida de cianose e ser filho de mãe não vacinada com dTpa. Sibilos e desconforto respiratório apresentaram alta sensibilidade para a identificação de VR. Na análise bivariada apresentaram maior chance de infecção por BP: menor idade (OR = 1,86), ausência de febre (OR = 4,9), não ser vacinado para coqueluche (OR = 4,4), leucocitose superior a 20.000/mm3 (OR = 5,4), linfocitose superior a 10.000/mm3 (OR = 4,0) e de infecção por VR: sibilos (OR = 4,33). Após o ajuste para confundidores, os maiores preditores para BP de forma independente foram: ausência de sibilos (OR =5,7) e leucocitose superior a 20.000/mm3 (OR = 5,38). O número de pacientes com codetecção não permitiu a análise comparativa de gravidade com aqueles com agente único. Em apenas um paciente o resultado da pesquisa viral positiva resultou em suspensão de macrolídeo. Conclusão: além da BP, os VR também foram etiologias frequentes nos lactentes com suspeita clínica de coqueluche, além de casos de codetecção de BP e VR. Foram identificadas características clínicas/laboratoriais sugestivas, porém não patognomônicas das etiologias identificadas o que corrobora a necessidade da pesquisa etiológica para VR, nessa situação clínicaIntroduction: Pertussis is a disease caused by Bordetella pertussis (BP), being more frequent and severe in infants less than one year old. After vaccine introduction, there was a reduction in the global incidence of the disease, but in the last ten years there was a resurgence. It may present less characteristically in infants, especially before the end of the vaccine scheme for the first year of life. The clinical picture in these patients may be similar to that of respiratory virus infections (VR), which are the most frequent etiologic agents in airway infections in this age group. Studies is necessary to evaluate the importance of RV research in infants with clinical suspicion of pertussis. Objectives: In infants with suspected pertussis: identify the prevalence of BP, VR and codetections; analyze and compare the clinical characteristics and evolution according to the identified etiology and analyze the impact of the etiological diagnosis on the use of macrolides. Methods: A prospective cohort study with children under one year of age hospitalized with suspected clinical pertussis between June 2014 and June 2016 and submitted to etiological research to identify BP (nasopharynx swab for culture and/or PCR) and VR (nasopharyngeal aspirate for indirect immunofluorescence). Clinical, demographic and evolution data were collected with the completion of a standardized clinical-laboratory protocol. Results: During the study period, 59 infants were analyzed. In 18 (30.5%) there was identification of BP, in 23 (39%) of some respiratory virus. In four (7%), there was BP detection and some RV. The virus most frequently identified was RSV (73%). The characteristics with greater sensitivity for the diagnosis of BP infection were cough followed by cyanosis and the mother\'s non-vaccinated dTpa. Wheezing and respiratory distress presented high sensitivity for RV identification. In the bivariate analysis they presented a greater chance of BP infection: lower age (OR = 1.86), absence of fever (OR = 4.9), not being vaccinated for pertussis (OR = 4.4), leukocytosis higher than 20,000/mm3 (OR = 5.4), lymphocytosis greater than 10,000/mm3 (OR = 4.0) and RV infection: wheezing (OR = 4.33). After adjustment for confounders, the largest predictors for BP independently were: no wheezing (OR = 5.7) and leukocytosis higher than 20,000/mm3 (OR = 5.38). The number of patients with codetection did not allow the comparative analysis of severity with those with single agent. In only one patient, the result of positive viral research resulted in macrolide suspension. Conclusion: In addition to BP, RVs were also frequent etiologies in infants with clinical suspicion of whooping cough, as well as cases of BP and VR codetection. Clinical/laboratory characteristics suggestive, but not pathognomonic, of the identified etiologies have been identified, which corroborates the need for etiological research for RV in this clinical situation
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Naninck, Thibaut. "Etude de l'infection par Bordetella pertussis dans un modèle de coqueluche chez le primate non-humain : Apports de l'imagerie in vivo." Thesis, Université Paris-Saclay (ComUE), 2018. http://www.theses.fr/2018SACLS526.
Full textAbstract:
La coqueluche est une pathologie due à la bactérie Bordetella pertussis qui touche les voies respiratoires des patients infectés causant toux, leucocytose, fièvre, et dont les symptômes peuvent aller jusqu’au décès chez les individus les plus à risque (nouveau-nés et enfants immunodéprimés en particulier). Ciblée par différents programmes vaccinaux depuis de nombreuses années, cette pathologie sévit à nouveau dans de nombreux pays développés où le nombre de cas augmente fortement depuis la fin des années 2000. Cette résurgence montre la nécessité de développer de nouvelles stratégies afin de comprendre les mécanismes de l’infection par B. pertussis. Dans ce contexte, la recherche préclinique apparaît comme essentielle pour comprendre la physiopathologie de la coqueluche. De nombreux modèles animaux ont été décrits pour l’étude de la coqueluche mais aucun de ces modèles ne permet de reproduire l’ensemble du spectre des symptômes cliniques de la pathologie, notamment la toux. Cependant, au cours des dernières années un modèle d’infection par Bordetella pertussis chez le jeune babouin a été développé aux Etats-Unis et permet de reproduire la pathologie observée chez l’homme, notamment concernant la toux et la transmission. Ce modèle semble ainsi très prometteur pour l’étude de la physiopathologie de la coqueluche.Cependant, de nombreuses inconnues subsistent dans ce modèle, notamment concernant la colonisation bactérienne et les interactions entre la bactérie et l’hôte. Nous avons ainsi cherché dans cette étude à évaluer d’une part l’impact de différents facteurs comme l’âge des animaux, la dose d’infection ainsi que la voie d’exposition sur la pathologie déclarée par les babouins suite à l’infection par la souche B1917 de B. pertussis afin de pouvoir proposer un parallèle avec les données cliniques disponibles. Nous avons également développé l’utilisation de techniques d’imagerie in vivo comme l’endomicroscopie confocale couplée à la bronchoscopie afin d’étudier la localisation et la cinétique de colonisation et certaines interactions du pathogène dans le tractus respiratoire inférieur au cours de la pathologie. Cette étude nous a ainsi permis d’approfondir les connaissances de physiopathologie de la coqueluche dans ce modèle babouin et consolidera cet outil précieux pour l’évaluation des futures stratégies de prévention contre cette pathologieWhooping cough, or pertussis, is a respiratory disease caused by Bordetella pertussis bacterial colonization of human airways. Main symptoms are cough, leukocytosis, fever and may even be lethal for some patients (e.g. newborn infants and immuno-deficient patients). Despite a good vaccination coverage worldwide against pertussis, whooping cough cases have been re-increasing in several developed countries in the past twenty years. This resurgence points out the crucial need to develop new control strategies and to better understand pertussis pathophysiology, notably using appropriate animal models. Numerous preclinical models including mice, rats, rabbits and swine have been described for B. pertussis infection studies. However, none of these models reproduce the full spectrum of clinical pertussis symptoms, especially cough. The recent baboon model of whooping cough described in the last few years in the US appears to be a very relevant model for pertussis pathophysiology studies as these animals reproduced all clinical symptoms as observed in humans including cough.However, many aspects of bacterial colonization and interactions with the host have yet to be described in this model.We have then evaluated diverse parameters such as animal age, the inoculum dose and the exposition route on the pathology symptoms and immune responses developed by baboons following B. pertussis B1917 strain inoculation in order to draw a parallel with human clinical data. We also developed in this model in vivo imaging techniques like confocal endomicroscopy coupled with bronchoscopy in order to evaluate bacterial colonization kinetics, localization and some interactions in the lower respiratory tract of infected baboons. Then, this study brought additional data on whooping cough physiopathology in this baboon model, which will be crucial for evaluating future prevention strategies against pertussis disease
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Apak, Aycan. "Assessment Of Immune Protective Capacity Of The Recombinant Iron-superoxide Dismutase (fesod) From Bordetella Pertussis." Master's thesis, METU, 2011. http://etd.lib.metu.edu.tr/upload/12613998/index.pdf.
Full textAbstract:
Whooping cough (pertussis) is a highly contagious acute respiratory disease caused by the strict human pathogen Bordetella pertussis, a gram-negative coccobacillus. The worldwide mass-vaccination was started in 1940s and to date, a number of whole-cell (Pw) and acellular pertussis vaccine (Pa) formulations were developed. Yet the current vaccines are incapable of providing sustained, lifelong immunity and eliminating subclinical infections, which pose a threat especially for unimmunized infants as well as adolescents and adults. Thus, finding new protein candidates with high immune protective capacities is necessary to enhance the clinical efficacy of current acellular pertussis (Pa) vaccines.
In this study, iron-superoxide dismutase (FeSOD) protein was investigated for its capacity of conferring protectivity as well as stimulating humoral and cellular responses against B. pertussis infection in a mouse model. For this purpose, sodB gene, which encodes iron-superoxide dismutase FeSOD protein, was amplified from the genomic DNA of the universal B. pertussis strain &lsquoTohama I&rsquo
and sequentially cloned to pGEM®
-T subcloning and pET-28a(+) expression vectors. Afterwards sodb/pET28a(+) construct was introduced to E. coli BL21(DE3) cells and the gene was overexpressed therein via IPTG induction. The expressed FeSOD protein was then purified by affinity chromatography and its previously reported immunogenicity was confirmed by Western blot. After filter-sterilization, the protein was adsorbed to alum [Al(OH)3] adjuvant and introduced to BALB/c twice at three weeks intervals intraperitoneally at a concentration of 20 &mu
g purified FeSOD protein/mouse. Another group of mice were immunized in tandem with heat-inactivated whole-cell suspension of B. pertussis. Ten days after the second immunization, mice were intranasally challenged with the local &lsquo
Saadet&rsquo
strain of B. pertussis. Next the lungs of groups of mice were excised, homogenized and plated as serial dilutions on days 5, 8 and 14 post-challenge, and viable lung CFU counts were carried out. Whole cell immunization conferred complete bacterial clearance following B. pertussis intranasal infection while FeSOD immunization failed to attain such protection. In addition to the protectivity assay, ELISA was performed to assess the humoral (i.e. IgG) immune response triggered upon FeSOD- and whole-cell immunizations and a statistically significant increase in anti-FeSOD IgG production was observed in FeSOD-immunized group. Finally, cellular immune response was tested via cytokine (IFN-&gamma
) assay, in which spleens of mice were excised, splenocytes were cultured and the level of IFN-&gamma
production upon FeSOD addition to the cultures was measured via ELISA. This test showed that whole-cell immunization triggered IFN-&gamma
production at significant levels while FeSOD-immunization did not
indicating the failure of alum-adsorbed FeSOD immunization in inducing cell-mediated immune response.