Introduction: Numerous studies have found associations between exposure to airborne particulate matter and respiratory morbidity and mortality. However, the varying composition, dependant on the different sources of particulate matter, and its effect on processes of inflammation and oxidative stress in the airways has not been completely elucidated. The use of airway biomarkers, fractional exhaled nitric oxide (FENO) and exhaled breath condensate (EBC), can provide valuable insight into processes of inflammation and oxidative stress in the airways. In these studies we sought to characterize the associations between airborne pollutant exposures and their sources and airway biomarkers. Methods:The study population of interest for Chapters III-V is a subset of children currently enrolled in a birth cohort under the Columbia Children's Center for Environmental Health. Chapter III refers to the validation in this population of a method for partitioning FENO contributions from the proximal and distal airways. Chapters IV and V refer to the implementation of this method in the study of ambient metals and residential proximity to relevant sources of particulate matter (PM) and black carbon (BC). Briefly, the children of African-American and Dominican mothers living in low-income NYC neighborhoods had FENO samples collected offline at constant flow rates of 50, 83 and 100 mL/sec at ages 9 and/or 11. Surrogate measures for bronchial flux NO (JNO) and alveolar (Calv) NO concentrations were estimated using a previously validated mathematical model. Wheeze in the last 12 months was assessed by the International Study of Asthma and Allergies in Childhood (ISAAC) questionnaire. Seroatopy was determined by specific IgE at age 9. For Chapter IV, in order to examine the associations between metal fractions of particulate matter and airway inflammation, ambient measures of Ni, V, Zn and Fe were obtained from a local central monitoring site and averaged over nine days based on three 24 hour measures every third day. Seroatopy was determined by specific IgE at age 7. For Chapter V, residential distance to a major road, truck route and bus stop density, area covered by major roads, stationary point sources (SPS), toxic release inventory sites (TRIS) and commercial buildings, and number of buildings burning residual oil within 250-meters of each child's home were determined. These variables were selected a priori as potential important sources or airborne PM and BC. For Chapter VI, the study population was comprised of seven- and eight-year-old children enrolled in an asthma case-control study in New York City. Seven day averages of domestic levels of particulate matter <2.5 microns (PM2.5), BC and environmental tobacco smoke (ETS) were measured. Urea and 8-isoprostane were measured by liquid chromatography tandem mass spectrometry (LC/MS/MS) in EBC collected during home visits. All data were analyzed with SPSS. Results: In our first study, children with seroatopy had significantly higher median JNO p<0.001) when compared to non-seroatopic children; however, median Calv was not significantly different between these two groups (p=0.644). Children with wheeze in the past year had significantly higher median Calv (p<0.001), but not JNO (295 vs. 165 pL/s, p=0.241) when compared with children without wheeze. In our second study, ambient V and Fe concentrations were associated positively with FENO50 (p=0.018, p=0.027). Ambient Fe was associated positively with JNO (p=0.017). Ambient Ni and V concentrations were associated positively with Calv (p=0.004, p=0.018 respectively). A stronger association of Ni concentrations with Calv was observed among the children with seroatopy. In our third study, there were no significant associations between any of the air pollution indicator variables and FENO parameters in adjusted models. In our final study, PM2.5and BC, but not ETS, were significantly associated with increases in 8-isoprostane (p<0.05 for both) after adjustment for covariates. In a co-pollutant model including PM2.5, BC and ETS, only BC remained a statistically significant predictor of 8-isoprostane. Discussion: Recent exposure to airborne pollutants was associated with increased levels of biomarkers of airway inflammation and oxidative stress measured in exhaled breath. The metal and BC fractions of PM might be more relevant to the understanding adverse respiratory outcomes related to air pollution exposure.

Highly pathogenic avian influenza H5N1, which is panzootic in poultry, continues to spread and becomes endemic in Asia, Africa, and Europe. It causes human disease with high fatality (about 60%) and continues to pose a pandemic threat. The pathological lesions associated with human H5N1 disease is Acute Respiratory Distress Syndrome (ARDS). The biological basis underlying the development of ARDS in human H5N1 disease remains controversial. Clinical, animal and in vitro studies suggested that the differences between H5N1 influenza viruss and low pathogenic influenza viruses in regard to viral replication, tissue tropism and cytokine dysregulation may contribute to disease pathogenesis. We previously found delayed onset of apoptosis in influenza H5N1 virus infected human peripheral blood monocyte-derived macrophages. This may allow a longer survival time for the virus in target cells for prolonged viral replication, which may contribute to the pathogenesis of H5N1 disease. As human bronchial and alveolar epithelial cells are target cells of influenza virus, I explored if influenza H5N1 and H1N1 virus infected human respiratory epithelial cells displayed differential apoptotic response and dissected the apoptotic pathways triggered by influenza virus infection. In this study, the apoptotic response in highly pathogenic influenza H5N1 viruses, A/HK/483/97 and A/Vietnam/1203/04, their precursor avian influenza H9N2 virus, A/Quail/HK/G1/97, and seasonal H1N1 virus, A/HK/54/98 infected primary human alveolar and bronchial epithelial cells was compared by TUNEL. A delayed onset of apoptosis in influenza H5N1 viruses and avian H9N2 virus infected alveolar epithelial cells was observed; the pattern was similar in bronchial epithelial cells. Concomitantly, by Western blotting, a delay in caspase 3 activation in H5N1 virus (A/HK/483/97) infected alveolar epithelial cells compared to H1N1 virus (A/HK/54/98) infected cells was shown. Also, influenza H5N1 and H1N1 virus induced apoptosis through both intrinsic and extrinsic pathways in human alveolar epithelial cells. Chemokine IP-10 was differentially up-regulated in influenza H5N1 virus infected alveolar epithelial cells, but its relationship to the delayed onset of apoptosis requires further studies. TRAIL, an upstream signaling molecule of extrinsic apoptotic pathway, mRNA was up-regulated in influenza H5N1 infected alveolar epithelial cells but not in influenza H1N1 infected cells. Using recombinant viruses, I showed that the 627 amino acid residue on PB2 of H5N1 virus and mutation of amino acids on 253 and 591 residues on PB2 of H9N2 virus contribute to the TRAIL upregulation. Immunohistochemical staining of physiologically relevant ex vivo model of human bronchus showed that influenza H5N1 (A/Vietnam/3046/04) and H9N2 (A/Quail/HK/G1/97) virus did not infect human bronchi as well as human H1N1 (A/HK/54/98) virus. Profiling of apoptosis related genes showed that TRAIL tends to be up-regulated in H5N1 virus infected bronchi ex vivo. This study demonstrated the delayed onset of apoptosis by H5N1 virus infected respiratory epithelial cells may be a mean for influenza virus to have prolonged replication within the human respiratory tract and contribute to disease severity. The results generated provide a robust research agenda, yielding critical information that elucidate molecular mechanisms, such as TRAIL up-regulation, that may contribute to the virulence and pathogenesis in human H5N1 disease. / HKU 3 Minute Thesis Award, 2rd Runner-up (2011) / published_or_final_version / Pathology / Master / Master of Philosophy

Background: Respiratory diseases are common worldwide, which are caused by various respiratory viruses. As symptoms caused by these viruses are similar, laboratory diagnosis is essential to distinguish the virus. Conventionally, respiratory viruses are isolated by cell culture with a panel of cell lines. However, handling of several cell lines is labour intensive, and the turnaround time of conventional culture is long. In previous study, the use of human colon adeno-carcinoma (Caco-2) in conventional culture was investigated. The study has proven that Caco-2 is generally susceptible to the eight common respiratory viruses, i.e. Adenovirus, Influenza A and B, Respiratory Syncytial virus, Parainfluenza virus 1, 2,3 and 4. As turnaround time of conventional culture is long; therefore, in this study, rapid shell vial culture using Caco-2 cells were evaluated. Moreover, the application of Caco-2 shell vial culture on recovering human metapneumovirus (hMPV) was also investigated. Materials and methods: This study consisted of four stages. First, recovery of viruses by conventional culture and shell vial culture of Caco-2 were compared. Specimens were added to conventional culture and shell vial simultaneously. For conventional culture, formation of CPE was examined daily and IF staining was performed when CPE was indicated; meanwhile, shell vial culture were incubated for seven days and stained with IF to detect infected cells. In stage two, the effect of incubating shell vial culture in rolling drum was investigated. Shell vials inoculated with the same specimen in duplicate were incubated in rolling drum and without rolling drum simultaneously. IF staining was performed in day 2, and results were obtained. For those which are IF negative in day 2, second shell vial was further incubated to seven days before harvest. In the next stage, a large batch of samples was used to evaluate on the use of Caco-2 shell vial culture in day 2 and day 7. Lastly, Caco-2 shell vial and conventional culture and LLC-MK2 conventional culture were tested for isolation of hMPV. Results: Compared to Caco-2 conventional culture, recovery rate of shell vial culture was elevated slightly. When experimenting on the effect of incubation in rolling drum, results showed that recovery rate was raised in shell vial with rolling drum in day 2, moreover, the percentage of positive cells were increased significantly (p value < 0.05). Furthermore, in the evaluation of Caco-2 shell vial in day 2 and day 7, 75% of samples were isolated in day 2 while 85% were recovered in day 7. Lastly, in the investigation on recovery of hMPV, 53%, 42% and 17% hMPV positive cases were isolated by Caco-2 shell vial, Caco-2 conventional culture and LLC-MK2 conventional culture respectively. Conclusion: First, although recovery rate by shell vial and conventional culture were similar, turnaround time was reduced from a week to a few days by shell vial culture. Therefore, Caco-2 shell vial culture is a more efficient than Caco-2 conventional culture in isolating respiratory viruses. The study also showed that incubation of shell vial in rolling drum able to increase the number of positive cells. Furthermore, in this study, Caco-2 cells were also shown to be more efficient in isolating hMPV when compare to LLC-MK2 cells. / published_or_final_version / Microbiology / Master / Master of Medical Sciences

No abstract available. / Thesis (M.Med.)-University of KwaZulu-Natal, 2004.

To investigate the possibility that malnutrition is an important factor that prolongs respiratory failure (PRF), I studied the effects of pharmacologic injections of recombinant human growth hormone (rhGH), an important anabolic stimulus, on nutritional and respiratory parameters in patients requiring mechanical ventilation for more than three days. Patients were excluded from consideration if dominating factors known to prolong ventilatory failure had not been stabilized. Over ten months, 106 patients in PRF were evaluated, but only six met the selection criteria. Three patients were randomized to receive standard nutritional support, and three into a group that received the equivalent nutrition plus 5 mg/day of rhGH for 14 days or until withdrawal of mechanical ventilation. Baseline characteristics of the selected patients were divergent as demonstrated by body mass indexes ranging from 14 to 42 (kg/m$ sp2),$ baseline maximal inspiratory pressures (PI$ sb{ max}$ from $-$15 to $-$70 cm H$ sb2$O, and Day 1 N balances from $-$13.5 to 1.2 g N/day. Despite increased plasma insulin-like growth factor-1 concentrations, the mean daily N balances of the rhGH-treated group were no better than the controls (1.3 $ pm$ 5.0 vs. 0.4 $ pm$ 2.6 g N/day; Mean $ pm$ SD), nor were there differences in PI$ sb{ max},$ level of ventilatory assistance required, and days to weaning. The persistence of respiratory failure in the overwhelming majority of patients in PRF appears to be due to factors already known to prevent weaning from mechanical ventilation. Even the carefully selected patients enrolled in the present study were insufficiently homogeneous or stable enough to allow proper testing of the experimental hypothesis.

A cross-sectional epidemiologic survey of the respiratory health status was conducted in the adult (15 years and older) Indian South African population resident in Lenasia, Johannesburg to study the distribution and determinants of respiratory symptoms, disease and lung function level. A slightly modified self-administered version of a standardised respiratory health questionnaire and a wedge spirometer was used. There were a high proportion of current smokers among men. Although women smoked less than men in other communities they nevertheless smoked on average more heavily than other Indian South African women. Indian men and women who smoked had a high prevalence of respiratory symptoms. The women also demonstrated an increased susceptibility to the effects of cigarette smoking when compared with women in other communities. Indians in this study had spirometric lung function levels that were lower than that recorded in recent studies in Blacks and Whites in South Africa. Respiratory symptoms, disease and lung function level were examined in a multiple logistic regression model which contained all the potential determinants recorded in the present study. Voluntary tobacco smoking, recent chest illnesses and any kind of heart trouble was associated with a significant risk for having most of the respiratory symptoms and diseases in men and women. In addition exposure to dust in the work environment, little or no exercise,>Std. 8 education a history of any kind of chest trouble and respiratory trouble before the age of 16 years was associated with an increased risk for having respiratory symptoms in men in this model. An increased risk for respiratory symptoms was demonstrated in women only with age. Age and standing height were the most important determinants of lung function level in men and women in the regression model. Dust exposure in the work environment was associated with a significantly lower lung function level in men. Alcohol consumption and a history of whooping cough was also independently associated with a lower lung function level in men but were of borderline significance. In women involuntary /passive tobacco smoke exposure and respiratory trouble before the age of 16 years were associated with a lower lung function level. Women who spent most of their lives in a rural area and those who had a university education had a higher lung function level. The deleterious effects of smoking on lung function were minimal in this study possibly because lung function was performed only in subjects in the 18-45 year age category. A "healthy smoker" effect was demonstrated in men. Men who ever smoked and were without cardiorespiratory symptoms had significantly higher lung function levels compared to men who never smoked and were without symptoms. / Thesis (M.D.)-University of Natal, Durban, 1992.

Introduction: Respiratory tract symptoms (RTS), common in athletes during heavy training and after events, result in impaired readiness for events and race times. Since the 1980’s exercise immunologists have investigated the aetiological factors surrounding the development of exercise induced RTS in order to develop effective preventative strategies. A number of theories have been put forward and explored, such as increased susceptibility to infection, ‘run-away’ inflammatory response and reactivation of prior viral infection. It has been suggested that the mechanisms producing exercise induced inflammation could potentiate allergic responses in sensitized individuals and recently allergic response has been proposed as a potential contributor to exercise induced RTS. Certainly allergic reactions can produce a range of respiratory symptoms; however the relationship between allergic sensitization, allergic reaction and the incidence of post-exercise RTS has not been well defined. Objectives: The primary objective of this study was to document the incidence of RTS for two weeks before and two weeks after a three-day trail run and relate these to the general systemic and salivary immunological profile as well as atopic status of the participants. The secondary objective was to validate the use of the Phadiatop® assay as a predictor of allergy-associated post-race RTS in trail runners. Study Design and Methods: The study formed part of a larger, descriptive field study examining the physiological responses of trail runners during the Three Cranes Challenge, a multi-day 95 km event divided into three stages, in Karkloof, KwaZulu-Natal. Outcome measures examined included self- reported RTS over a 31 day period (pre, during and post race), as well as pre-race Phadiatop® status, salivary IgA (sIgA) concentrations and changes in concentrations of serum IgE (sIgE), cortisol, high sensitivity C-Reactive Protein (hs-CRP) and differential leukocyte counts. The haematological and salivary parameters were obtained at 8 time points before, during and after the event. A convenience sample of 22 individuals was used and two separate analyses were conducted on the data. The inclusion criteria of the first analysis were met by 14 participants. In this analysis, the incidence of RTS was related to each participant’s general immunological profile. Sixteen of the subjects met the inclusion criteria for the second analysis, in which their Phadiatop® status was related to their sIgE and blood eosinophil and basophil concentrations in order to establish the validity of the Phadiatop® assay in predicting the development of allergy–associated postexercise RTS in trail runners. Results: In the first analysis, 78.6 % (n=11) of subjects met the criteria for positive diagnosis of upper respiratory symptoms (URS) during the two week post-race period. In four subjects (36.4 %), URS appeared to be of inflammatory origin, but these were not linked to systemic markers of an allergic response. Of the URS positive subjects, six (54.5 %) presented with markers of infection, three (27.3 %) with markers of a de novo infection and three (27.3%) with a profile suggestive of reactivation of previous infection. Of those presenting with markers of infection 66.7 % (n=4) had concomitantly elevated levels of IgE suggestive of allergic response. There was, however, no evidence of isolated allergic reaction independent of other causes amongst the symptomatic subjects. In the second analysis, 75% (n=12) of runners presented with post-race RTS and seven of these were Phadiatop® positive. In four of the Phadiatop® positive RTS subjects, symptoms appeared to be of allergic origin. Although total IgE concentrations were significantly higher (p< 0.01) in Phadiatop® positive group, there was no significant difference between the eosinophil and basophil concentrations or post-race RTS of the positive and negative groups (p>0.05). Of the four subjects who did not develop RTS, three were Phadiatop® positive. Conclusion: Respiratory tract symptoms in trail runners have a multi-factorial aetiology. A link between concurrent markers of an allergic response and infection is common in symptomatic trail runners. The Phadiatop® assay does not accurately predict the incidence of allergic postexercise RTS in trail runners. / Thesis (M.Med.Sc.)-University of KwaZulu-Natal, Westville, 2012.

In order to evaluate the respiratory health status of spray painters exposed to paints containing hexamethylene diisocyanates (HDI) and to obtain more insight into the relationship between occupational exposures to isocyanates and chronic obstructive airway diseases, a follow up study on 33 of an original cohort of 40 randomly selected workers was undertaken. The original investigation was conducted by the author in 1989. The subjects were studied using a standardised American Thoracic Society (ATS) approved respiratory health questionnaire, baseline pre and post shift spirometry and ambulatory peak flow monitoring. Bronchial hyperresponsiveness tests using histamine (PC20) were performed. Immunological tests including IgE, RAST (HDI), and house dust mite evaluations were also made. The subjects were stratified into exposed (n=20), partially exposed (n=5) and no longer exposed (n=7) groups. One subject was excluded from the group analysis because of his indeterminate isocyanate exposure. Warehouse assistants (n=30) in a non-exposed occupation were used as controls. The worker's compliance with safety regulations and the employers provision of safety requirements was assessed by means of a questionnaire. The environmental conditions in the workplace were measured by the evaluation of the isocyanate concentrations at the worker's breathing zone. Spray booth efficiency was measured using measurements of airflow velocities and airflow patterns within the booth. Longitudinal changes in respiratory health status was assessed by comparison with baseline data studied in 1989. The exposed group showed the largest mean cross-shift declines of 297 ml (± 83.8) in forced expiratory volume in one second (FEV1). The decline in the partially exposed group was 282 ml (± 102.7) and 54 ml (± 140) in the no longer exposed group. The results of the first study, when compared with the second study, showed a mean cross-shift decline in FEV1 of 130.5 ml. (± 203) (p=0.0002) and 297ml. (± 323) (p=0.0001) respectively. Furthermore, of the spray painters examined, 10 (25%) showed clinically significant cross-shift declines in FEV1 viz. decreases >250 ml in the first study (n=40) compared with 9 (45%) in the second study (n=33). In contrast to the HDI exposed spray painters, a closely matched control group (n=30) showed a mean cross-shift increase in FEV1 of 17.4 ml ( ± 63.04). Only 2 subjects had a diagnosis of asthma which was made in childhood and not related to occupation. The mean annual baseline decline in FEV1 was greatest in the exposed group 41.25 ml (25% showed a decline greater than >90 ml per annum). These values exceeded the predicted annual declines for both smokers and non smokers due to age. The decline in the no longer exposed group was 7.85 ml per annum. Immunological tests showed no correlation with declines in FEV1 . This study demonstrates the difficulties in correlating immunological status with clinical and lung function findings in workers exposed to HDI, as a means of predicting occupational asthma. Although measurements in cross-shift declines in FEV1 appear to be a suitable predictor of occupational asthma, in some cases it was found that the forced expiratory flow rate (FEF 25-75 %) was a more sensitive predictor of early changes in the small airways. The mean isocyanate concentration in the spray painter's breathing zone was 14.65 mg/m3 (±12.219), exceeding the current South African Occupational Exposure Limit - Control Limit (OEL-CL) of 0.07 mg/m3 for isocyanates. Fifty per cent of the subjects suffered from eye irritation and 40% had dermatitis of the hand. This was expected since none of the spray painters wore goggles or gloves. Whilst no subject had evidence of clinical asthma related to spray painting, a large proportion demonstrated significant cross- shift changes in lung function implying short- term adverse effects of exposure. In addition longitudinal declines in lung function which was worse in those who continued spray painting in the follow-up study, is of major concern. The lack of cases of clinical or occupational asthma may be due to the healthy worker effect. Recommendations include, routine spirometric lung function testing of all spray painters, the use of high volume-low pressure spray guns and the wearing of positive pressure airline masks complying with the South African Bureau of Standards (SABS) safety standard. In terms of current legislation it was further recommended that spray booths be regularly monitored, including the measurement of HDI concentrations, airflow velocities and airflow patterns within the booth and the implementation and enforcement of stricter control measures. Workers demonstrating excessive declines in both cross-shift and longitudinal spirometry, require special attention. / Thesis (M.Med.Sc.)-University of Natal, 1997.

Orientador: Irma de Godoy / Banca: Hugo Hyung Bok Yoo / Banca: Fabio de Oliveira Pitta / Banca: Maria Christina L. O. Machado / Banca: Alberto Cukier / Resumo: Nos últimos anos, vários estudos avaliaram os marcadores da doença associados à freqüência de exacerbação, hospitalização, readmissão e mortalidade em pacientes portadores de doença pulmonar obstrutiva crônica (DPOC). Entretanto, estudos que avaliaram os marcadores associados à ocorrência de exacerbação em pacientes ambulatoriais são limitados. Assim, o objetivo deste estudo foi identificar os fatores de predição da ocorrência de exacerbação no período de um ano em 120 pacientes com DPOC atendidos no Ambulatório de Pneumologia da Faculdade de Medicina de Botucatu - Unesp. Os pacientes tiveram o diagnóstico de DPOC confirmado e foram submetidos às seguintes avaliações: espirometria pré e pós-broncodilatador, composição do corpo (antropometria e bioimpedância), qualidade de vida por meio do Saint George’s Respiratory Questionnaire (SGRQ), intensidade da dispnéia por meio da escala modificada Medical Research Council (MMRC) e do índice basal de dispnéia (BDI) e tolerância ao exercício (distância percorrida em 6 minutos – DP6). Em seguida foi calculado o índice BODE de acordo com os pontos de corte do volume expiratório no primeiro segundo (VEF1), do índice de massa do corpo, do MMRC e da DP6. Durante o período de acompanhamento de um ano, 60 pacientes (50%) apresentaram pelo menos um episódio de exacerbação da doença e, em conseqüência da agudização, 25 pacientes foram hospitalizados. Comorbidades extra-pulmonares foram causa de hospitalização em oito pacientes e de óbito em cinco pacientes. Na avaliação inicial, os pacientes que exacerbaram tinham maior comprometimento da função pulmonar e da troca gasosa, valores mais elevados do índice BODE e maior proporção de pacientes com DPOC III e IV. Além disso, apresentavam menores valores de DP6, maior sensação de dispnéi... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Markers of disease severity have been associated with mortality, occurrence and frequency of hospitalization and readmission due disease exacerbation in chronic obstructive pulmonary disease (COPD) patients. However, information about predictor factors for the occurrence of exacerbation in ambulatory COPD patients is scarce. Therefore, the aim of the present study was to identify predictor factors for the occurrence of exacerbation in 120 patients with COPD followed during one year in the outpatient clinic at Universidade Estadual Paulista (UNESP, Paulista State University) School of Medicine at Botucatu, located in the State of São Paulo, Brazil. The patients had the diagnosis of COPD confirmed and underwent to the following evaluations: pre- and postbronchodilator spirometry, body composition (anthropometry and bioimpedance), health-related quality of life (Saint George’s Respiratory Questionnaire -SGRQ), dyspnea scores (Medical Research Council –MMRC and basal dispnea index -BDI) and exercise tolerance (6MWD). Bode index was calculated taking in consideration the cutt off points for forced volume in the first second (FEV1), body mass index, MMRC and 6MWD. During the followup period 60 patients (50%) presented at least one exacerbation episode and, as consequence, 25 patients were hospitalized. Eight patients were hospitalized and five died due to non-pulmonary comorbidities. At baseline, patients with exacerbations during the follow-up period presented lower values of airway obstruction indexes and of arterial blood gases and higher values of BODE score and proportion of COPD patients class III and IV. In addition, the values of 6MWD were lower, dyspnea sensation was higher and the health- related quality of life was more deteriorated in these patients. No significative associations were found between gender, corticosteroid use... (Complete abstract click electronic access below) / Doutor

No description available.