Valores de referência para a difusão do monóxido de carbono (fator de transferência) em uma amostra brasileira da raça branca / Reference values for the carbon monoxide diffusion (transfer factor) in a brazilian sample of white race

RESUMO Objetivo Derivar valores de referência em adultos brancos para a difusão do monóxido de carbono (DCO) em uma amostra de diferentes locais do Brasil por um mesmo modelo de equipamento (Sensormedics) e comparar os resultados com os derivados pelas equações de Crapo, Miller e Neder, e da proposta pelo Global Lung Initiative (GLI). Métodos Os testes foram realizados de acordo com as normas sugeridas pela ATS/ERS, em 2005, em seis cidades brasileiras, em 120 voluntários adultos de cada sexo, não fumantes, sem anemia referida e sem doenças pulmonares ou cardíacas. Os previstos foram derivados por regressões lineares e as diferenças entre os valores previstos por alguns autores e os observados no estudo atual foram calculadas. Resultados Nos homens, a idade variou de 25 a 88 anos e a estatura, de 156 a 189 cm. Nas mulheres, a idade variou de 21 a 92 anos e a estatura, de 140 a 176 cm. A DCO se correlacionou de maneira significativa positivamente com a estatura e negativamente com a idade. Os valores previstos pelas equações de Crapo, Neder e Miller foram maiores em comparação aos obtidos pelo estudo atual (p<0,01) em ambos os sexos. Nos homens, os valores não diferiram quando comparados aos calculados pelo GLI (p=0,29); nas mulheres, os valores derivados pelo GLI foram levemente maiores: 0,99 mL/min/mmHg (p<0,01). Conclusões Novos valores previstos para a DCO foram derivados em uma amostra de adultos brancos no Brasil. Os valores previstos são semelhantes aos compilados pelas equações GLI e diferem de equações propostas anteriormente.

ABSTRACT Objective To derive reference values from white race adults, for DCO in a sample from different sites in Brazil, through the same equipment model (Sensormedics), and compare the results with the derivatives from Crapo, Miller, Neder equations and from the Global Lung Initiative (GLI) proposal. Methods The tests were performed according to the norms suggested by ATS/ERS in 2005 in six Brazilian cities, with 120 adult volunteers of each gender, non-smokers, without referred anemia and without lung or cardio diseases. The expected values were derived from linear regressions and the differences between the values forecasted by some authors and the ones observed in the current study were calculated. Results Among men, the age varied between 25 and 88 years old, and the height varied between 140 and 176 cm. DCO was correlated significantly and positively with the height and negatively with the age. The values forecasted by Crapo, Neder, and Miller equations were higher in comparison with the ones obtained by the current study (p<0.01) in both genders. Among men, the values did not differ when compared to the ones calculated by GLI (p=0.29); among women, the values derived by GLI were slightly higher: 0.99 ml/min/mmHg (p<0.01). Conclusion new values forecasted for DCO were derived in a sample of white adults in Brazil. The forecasted values are similar to the ones complied by GLI equations and differ from the previously proposed equations.

Activation of locus coeruleus heme oxygenase-carbon monoxide pathway promoted an anxiolytic-like effect in rats

The heme oxygenase-carbon monoxide pathway has been shown to play an important role in many physiological processes and is capable of altering nociception modulation in the nervous system by stimulating soluble guanylate cyclase (sGC). In the central nervous system, the locus coeruleus (LC) is known to be a region that expresses the heme oxygenase enzyme (HO), which catalyzes the metabolism of heme to carbon monoxide (CO). Additionally, several lines of evidence have suggested that the LC can be involved in the modulation of emotional states such as fear and anxiety. The purpose of this investigation was to evaluate the activation of the heme oxygenase-carbon monoxide pathway in the LC in the modulation of anxiety by using the elevated plus maze test (EPM) and light-dark box test (LDB) in rats. Experiments were performed on adult male Wistar rats weighing 250-300 g (n=182). The results showed that the intra-LC microinjection of heme-lysinate (600 nmol), a substrate for the enzyme HO, increased the number of entries into the open arms and the percentage of time spent in open arms in the elevated plus maze test, indicating a decrease in anxiety. Additionally, in the LDB test, intra-LC administration of heme-lysinate promoted an increase on time spent in the light compartment of the box. The intracerebroventricular microinjection of guanylate cyclase, an sGC inhibitor followed by the intra-LC microinjection of the heme-lysinate blocked the anxiolytic-like reaction on the EPM test and LDB test. It can therefore be concluded that CO in the LC produced by the HO pathway and acting via cGMP plays an anxiolytic-like role in the LC of rats.

Elevated Erythrocyte Sedimentation Rate Is Predictive of Interstitial Lung Disease and Mortality in Dermatomyositis: a Korean Retrospective Cohort Study

Interstitial lung disease (ILD) is a major cause of death in patients with dermatomyositis (DM). This study was aimed to examine the utility of the erythrocyte sedimentation rate (ESR) as a predictor of ILD and prognostic marker of mortality in patients with DM. One hundred-and-fourteen patients with DM were examined, including 28 with clinically amyopathic DM (CADM). A diagnosis of ILD was made based on high resolution computed tomography (HRCT) scans. The association between elevated ESR and pulmonary impairment and mortality was then examined. ILD was diagnosed in 53 (46.5%) of 114 DM patients. Cancer was diagnosed in 2 (3.8%) of 53 DM patients with ILD and in 24 (92.3%) of those without ILD (P or = 30 mm/hour had significantly higher mortality than those with ESR < 30 mm/hour (P = 0.002, log-rank test). Patients with a persistently high ESR despite immunosuppressive therapy was associated with higher mortality than those with a normalized ESR (P = 0.039, log-rank test). Elevated ESR is associated with increased mortality in patients with DM due to respiratory failure. Thus, monitoring ESR should be an integral part of the clinical care of DM patients.

Acute stress-induced antinociception is cGMP-dependent but heme oxygenase-independent

Endogenous carbon monoxide (CO), which is produced by the enzyme heme oxygenase (HO), participates as a neuromodulator in physiological processes such as thermoregulation and nociception by stimulating the formation of 3′,5′-cyclic guanosine monophosphate (cGMP). In particular, the acute physical restraint-induced fever of rats can be blocked by inhibiting the enzyme HO. A previous study reported that the HO-CO-cGMP pathway plays a key phasic antinociceptive role in modulating noninflammatory acute pain. Thus, this study evaluated the involvement of the HO-CO-cGMP pathway in antinociception induced by acute stress in male Wistar rats (250-300 g; n=8/group) using the analgesia index (AI) in the tail flick test. The results showed that antinociception induced by acute stress was not dependent on the HO-CO-cGMP pathway, as neither treatment with the HO inhibitor ZnDBPG nor heme-lysinate altered the AI. However, antinociception was dependent on cGMP activity because pretreatment with the guanylate cyclase inhibitor 1H-[1,2,4] oxadiazolo [4,3-a] quinoxaline-1-one (ODQ) blocked the increase in the AI induced by acute stress.

Dificultades en la elección de una ecuación de referencia para la interpretación de los resultados de capacidad de difusión de monóxido de carbono / Difficulties in the selection of a reference equation for the interpretation of the results of DLCO

Introduction: In the absence of national reference equations, the ATS recommends comparing the results of the diffusion capacity of healthy subjects in a representative sample of the population with international equations and choosing among them, the one that provides the lowest sum of the residues. Objective: To compare reference equations available in the literature and identify which best meets the selection criteria. Methods: We reviewed 10 reference equations, for each one we calculated the sum of the residues for a sample of 71 healthy subjects and described the characteristics that affect the variability of each one. Results: Since 1985 we have used the single breath method. Only Thompson et al prediction equations 2008 were obtained with instantaneous reading of CO. Equations with the lowest sum of residues (Miller, Roca and Cotes) include smokers and former smokers. Conclusions: We need an equation in non-smokers with methodology that ensures low variability.

Introducción: En ausencia de ecuaciones de referencia nacionales, la ATS recomienda comparar los resultados de capacidad de difusión de monóxido de carbono de sujetos sanos en una muestra representativa de la población, con ecuaciones internacionales y escoger entre estas, aquella que presente la menor suma de los residuos. Objetivo: Comparar las ecuaciones de referencia disponibles en la literatura e identificar cuál cumple mejor los criterios de selección. Método: Revisamos 10 ecuaciones de referencia; calculamos la suma de los residuos de cada una de ellas para una muestra de 71 sujetos sanos y describimos las características que inciden en la variabilidad de cada una. Resultados: Desde 1985 se ha utilizado el método de respiración única. Sólo las ecuaciones de Thompson y cols. 2008fueron obtenidas con lectura instantánea. Las ecuaciones que presentan menor suma de residuos (Miller, Roca y Cotes) incluyen fumadores y ex fumadores. Conclusiones: Es necesaria una ecuación nacional en sujetos sanos no fumadores, con metodología que asegure baja variabilidad.

Central chemoreceptors and neural mechanisms of cardiorespiratory control

The arterial partial pressure (P CO2) of carbon dioxide is virtually constant because of the close match between the metabolic production of this gas and its excretion via breathing. Blood gas homeostasis does not rely solely on changes in lung ventilation, but also to a considerable extent on circulatory adjustments that regulate the transport of CO2 from its sites of production to the lungs. The neural mechanisms that coordinate circulatory and ventilatory changes to achieve blood gas homeostasis are the subject of this review. Emphasis will be placed on the control of sympathetic outflow by central chemoreceptors. High levels of CO2 exert an excitatory effect on sympathetic outflow that is mediated by specialized chemoreceptors such as the neurons located in the retrotrapezoid region. In addition, high CO2 causes an aversive awareness in conscious animals, activating wake-promoting pathways such as the noradrenergic neurons. These neuronal groups, which may also be directly activated by brain acidification, have projections that contribute to the CO2-induced rise in breathing and sympathetic outflow. However, since the level of activity of the retrotrapezoid nucleus is regulated by converging inputs from wake-promoting systems, behavior-specific inputs from higher centers and by chemical drive, the main focus of the present manuscript is to review the contribution of central chemoreceptors to the control of autonomic and respiratory mechanisms.

Breath carbon monoxide levels in different forms of smoking.

Background and Objectives. Bidi, cigarette, hookah and chillum are common modes of tobacco smoking in India. Many people consider hookah and chillum smoking less toxic because smoke is filtered through water or wet cloth. We evaluated the toxicity of tobacco smoking by measuring end-tidal carbon monoxide (eCO) levels after various modes of smoking. Methods. Eighteen healthy smokers who smoked bidi, cigarette, hookah and chillum on six days were studied. They smoked one bidi, one cigarette, five minutes hookah, one serve (15 minutes) hookah, five minutes chillum and one serve (15 minutes) chillum on six days randomly. The eCO values were measured before initiation of smoking and for a period of one hour after the smoking session. Increase in eCO values in comparison to baseline after different modes of smoking was compared. Results. In comparison to baseline, mean eCO levels were raised by 4.94 (0.96) parts per million (ppm) immediately and 4.17 (1.07) ppm 60 minutes after cigarette smoking. Bidi smoking caused slightly less increase in mean eCO levels (3.17 [0.82]). One serve of hookah and chillum smoking caused elevation of mean eCO values by almost eight-folds higher than that of cigarette smoking. Five minutes of smoking with hookah (22.18 [5.29]) and one serve of hookah (33.0 [8.76]) and chillum (40.14 [12.73]) caused significantly higher values of mean increase in eCO in comparison to cigarette smoking (p<0.001). Conclusion. With regard to eCO levels, hookah and chillum smoking are much more toxic than cigarette smoking.

Risk Factors of Postoperative Pneumonia after Lung Cancer Surgery

The purpose of this study was to investigate risk factors of postoperative pneumonia (POP) after lung cancer surgery. The 417 lung cancer patients who underwent surgical resection in a tertiary referral hospital were included. Clinical, radiological and laboratory data were reviewed retrospectively. Male and female ratio was 267:150 (median age, 65 yr). The incidence of POP was 6.2% (26 of 417) and in-hospital mortality was 27% among those patients. By univariate analysis, age > or = 70 yr (P or = 4.2 hr (P = 0.043), intraoperative red blood cells (RBC) transfusion (P = 0.004), presence of postoperative complications other than pneumonia (P = 0.020), forced expiratory volume in 1 second/forced vital capacity (FEV1/FVC) or = 0.15 mg/dL (P = 0.001) were related with risk of POP. Multivariate analysis showed that age > or = 70 yr (OR = 3.563, P = 0.014), intraoperative RBC transfusion (OR = 4.669, P = 0.033), the presence of postoperative complications other than pneumonia (OR = 3.032, P = 0.046), and FEV1/FVC < 70% (OR = 3.898, P = 0.011) were independent risk factors of POP. In conclusion, patients with advanced age, intraoperative RBC transfusion, postoperative complications other than pneumonia and a decreased FEV1/FVC ratio have a higher risk for pneumonia after lung cancer surgery.

Prevalência de tabagismo ativo e passivo em uma população de asmáticos / Prevalence of active and passive smoking in a population of patients with asthma

O tabagismo causa intensa reação inflamatória nas vias aéreas e, em asmáticos, está associado com piores desfechos clínicos. O objetivo desse estudo foi determinar a prevalência de tabagismo ativo e passivo em uma população de asmáticos. A amostra de pacientes com asma (n = 100) consistiu em 47 não-fumantes, 33 ex-fumantes e 3 fumantes ativos. A maioria dos pacientes tinha asma moderada ou grave. A média de CO exalado foi de 9,34 ppb nos tabagistas atuais, 4,19 ppb nos fumantes passivos e 3,98 ppb tanto nos não-fumantes quanto nos ex-fumantes. Concluímos que a prevalência da exposição à fumaça do tabaco é alta entre asmáticos.

Smoking causes an intense inflammatory reaction in the airways and is associated with worse clinical outcomes in patients with asthma. The objective of this study was to determine the prevalence of active and passive smoking in a population of patients with asthma. The sample of asthma patients (n = 100) consisted of 47 nonsmokers, 33 former smokers, 17 passive smokers and 3 active smokers. Most had moderate or severe asthma. Mean exhaled CO was 9.34 ppb in smokers, 4.19 ppb in passive smokers, 3.98 ppb in nonsmokers and 3.98 ppb in former smokers. We conclude that the prevalence of exposure to tobacco smoke is high among asthma patients.

Antinociception synergy between the peripheral and spinal sites of the heme oxygenase-carbon monoxide pathway

We have shown that the peripheral and spinal cord heme oxygenase (HO)-carbon monoxide (CO)-soluble guanylate cyclase-cGMP pathways play an important role in antinociception in the rat experimental formalin model. Our objective was to determine if there is synergism between peripheral (paw) and spinal HO-CO pathways in nociception. Rats were handled and adapted to the experimental environment for a few days before the formalin test, in which 50 µL of a 1 percent formalin was injected subcutaneously into the dorsal surface of the right hind paw. The animals were then observed for 1 h and the frequency of flinching behavior was taken to represent the nociceptive response. Thirty minutes before the test, rats were pretreated with intrathecal injections of the HO inhibitor, zinc deuteroporphyrin 2,4-bis glycol (ZnDPBG) or heme-lysinate, which is a substrate of the HO pathway. The paw treatments took place 20 min before the test. Low doses of ZnDPBG did not increase nociception, while a low heme-lysinate dose did not change flinching behavior after paw or spinal injections. Combined subactive spinal (50 nmol) and peripheral (40 nmol) low doses of ZnDPBG induced hypernociception (increase of 80 percent in the first and 25 percent in the second phase flinching), whereas combined spinal-peripheral heme-lysinate (50 and 30 nmol) led to second phase antinociception (40 percent reduction in flinching). These findings suggest a synergy between the peripheral and spinal HO-CO pathways. Local activation of the HO system probably regulates the nociception initiation in peripheral tissue and participates in buffering the emerging nociceptive signals at the peripheral and spinal sites of action. In short, an antinociceptive synergy exists between peripheral and spinal HO pathways, which may reduce the doses required and side effects.

Compromiso de la función pulmonar en pacientes con fíbrosis pulmonar idiopática / Pulmonary function impairment in patients with idiopathic pulmonary fibrosis

Idiopathic pulmonary fibrosis (LPF) is the most frequent of idiopathic interstitial pneumonias. The most sensitive lung function test for its diagnosis is a low DLCO register. Objective: To analyze LPF's clinical features and to establish its functional disability by DLCO at the time of diagnosis and also to correlate DLCO with 6-minute walked distance test (DR6), saturation pre and post DR6 and force a vital capacity (FVC). Methods: We studied retrospective data from 31 patients with LPF, analyzing personal and family history. We also determined pulmonary function parameters: DLCO (stratified in 4 groups: normal, mild, moderate and severe impairment), FVC, DR6 with pre and post oxygen saturation. Functional impairment according to DLCO was severe in 7patients and moderate in 19 out of the 31 patients. Results: An inverse association was observed between DLCO impairment and oxygen saturation levéis in rest, exercise and the difference between them (p < 0.001). Conclusion: Oxygen saturation level in exercise and rest, as well as the difference between them, have a good correlation with DLCO for LPF diagnosis.

La fibrosis pulmonar idiopática (FPL) es la más común de las neumonías intersticiales idiopáticas. De los exámenes de función pulmonar, la DLCO es la más sensible para su diagnóstico. Objetivo: Revisar las características de los pacientes con FPL y determinar su compromiso funcional según DLCO. Correlacionar la DLCO con el test de caminata de 6 minutos (DR6), saturación pre y post DR6 y CVF. Método: Estudio retrospectivo de 31 pacientes con FPL, se analizó antecedentes clínicos, familiares e índices de función pulmonar: DLCO (estratificada en normal, disminución leve, moderada y severa), CVF, DR6 y Sa0(2) antes y después de DR6. Resultados: El compromiso funcional observado según DLCO fue grave en 7 pacientes y moderado en 19. Hubo asociación inversa entre gravedad según DLCO, saturación en reposo y en ejercicio y con la diferencia de saturación en reposo y ejercicio (p < 0,001). Conclusión: La saturación de oxígeno en reposo y ejercicio, así como la diferencia entre ellas, tuvieron buena correlación con DLCO para el diagnóstico de FPL.

Correlação dos achados tomográficos com parâmetros de função pulmonar na fibrose pulmonar idiopática em não fumantes / Correlation of tomographic findings with pulmonary function parameters in nonsmoking patients with idiopathic pulmonary fibrosis

OBJETIVO: Correlacionar os achados tomográficos com os parâmetros de função pulmonar em portadores de fibrose pulmonar idiopática (FPI). MÉTODOS: Foi realizado um estudo de corte transversal, em que foram avaliados 30 pacientes não tabagistas, portadores de FPI. Utilizando um sistema de escore semiquantitativo, os seguintes achados na tomografia computadorizada de alta resolução (TCAR) foram quantificados: extensão total da doença intersticial (Tot), infiltrado reticular e faveolamento (Ret+Fav), e opacidade em vidro fosco (Vif). As variáveis funcionais foram mensuradas através de espirometria, técnica de oscilações forçadas (TOF), método da diluição com hélio e método da respiração única para medir a capacidade de difusão do monóxido de carbono (DLCO). RESULTADOS: Dos 30 pacientes estudados, 18 eram mulheres e 12 eram homens, com média de idade de 70,9 anos. Foram encontradas correlações significativas de Tot e Ret+Fav com as medidas de capacidade vital forçada (CVF), capacidade pulmonar total (CPT), DLCO e complacência dinâmica do sistema respiratório (correlações negativas), e de Vif com volume residual/CPT (correlação positiva). A relação fluxo expiratório forçado entre 25 e 75 por cento da CVF/CVF (FEF25-75 por cento/CVF) correlacionou-se positivamente com Tot, Ret+Fav e Vif. CONCLUSÕES: Em portadores de FPI, as medidas de volume, difusão e complacência dinâmica são as variáveis fisiológicas que melhor refletem a extensão da doença intersticial na TCAR.

OBJECTIVE: To correlate tomographic findings with pulmonary function parameters in patients with idiopathic pulmonary fibrosis (IPF). METHODS: A cross-sectional study was carried out, in which 30 nonsmoking patients with IPF were evaluated. Using a semiquantitative scoring system, the following high-resolution computerized tomography (HRCT) findings were quantified: total interstitial disease (TID), reticular abnormality/honeycombing, and ground-glass opacity (GGO). The functional variables were measured by spirometry, forced oscillation technique (FOT), helium dilution method, as well as the single-breath method of measuring diffusion capacity of the lung for carbon monoxide (DLCO). RESULTS: Of the 30 patients studied, 18 were female, and 12 were male, with a mean age of 70.9 years. We found that TID and reticular abnormality and honeycombing correlated significantly (negative correlations) with the measurements of forced vital capacity (FVC), total lung capacity (TLC), DLCO, and dynamic respiratory compliance were found, as well as that GGO correlated significantly (and positively) with residual volume/TLC. The ratio of forced expiratory flow between 25 and 75 percent of FVC to FVC (FEF25-75 percent/FVC) correlated positively with TID, reticular abnormality/honeycombing, and GGO. CONCLUSION: In IPF patients, the measurements of volume, diffusion, and dynamic compliance are the physiological variables which best reflect the extent of the interstitial disease on HRCT scans.

Compromiso pulmonar en esclerosis sistémica / Lung involvement in systemic sclerosis

El objetivo del estudio fue determinar las características clínicas de los pacientes con esclerodermia y compromiso pulmonar y evaluar si existen factores clínicos predictores de mayor riesgo de enfermedad intersticial. Se estudiaron en forma retrospectiva 40 pacientes con esclerodermia. Fueron divididos en 2 grupos: capacidad de difusión del monóxido de carbono (DLCO) normal (n = 22) y DLCO disminuida (n = 18, 45%). Los pacientes con DLCO disminuida no fueron diferentes en edad (51.1 más o menos 13.5 vs. 53.5 más o menos 9.3 años, p = 0.5182), sexo (varones 13.6%, p = 0.6088 ), presencia de Raynaud (86.6% vs. 85%, p = 0.6272), síndrome de ojo seco (6.2% vs. 10.5%, p = 1.0000) prevalencia de enfermedad difusa (94.1% vs. 83.3%, p = 0.6026) o de dilatación esofágica. El tiempo de evolución de la enfermedad no fue diferente. La sensibilidad de la disnea para detectar una DLCO alterada fue 46.6% con una especificidad del 90% y la de la caída de la saturación de O2 (SaO2) del 71.4% y 80% respectivamente. Los pacientes con DLCO baja tuvieron mayor prevalencia de anticuerpos anti-Scl 70 positivos (5/9 vs. 0/11, p = 0.0081) y de incapacidad ventilatoria restrictiva aunque en 56.7% de los pacientes con DLCO disminuida la capacidad pulmonar total (CPT) era normal. La presencia de hipertensión pulmonar medida por ecocardiograma Doppler fue idéntica (11/13 vs. 10/11, p = 1.0000). Los pacientes con DLCO disminuida tuvieron una prevalencia muy superior de tomografía computada de tórax con evidencias de compromiso intersticial (82.3% vs. 5.8%, p menor o igual a 0.0001). En conclusión, nuestros datos sugieren que la disminución de la DLCO es un hallazgo, muy frecuentemente asociado a TAC de tórax con compromiso intersticial y que no hay variables clínicas que permitan predecir su anormalidad.

The objective of this study was to determine clinical predictors of interstitial lung disease in patients with systemic sclerosis (SSc) and pulmonary involvement as defined by presence of a decreased diffusing capacity for carbon monoxide (DLCO). Forty subjects with SSc were retrospectively evaluated. Patients were categorized according to their level of DLCO (less than or more than or equal to 80% of predicted). Sensitivity of dyspnea to detect a decreased DLCO was 46.6% and specificity 90%, whereas oxygen desaturation showed a sensitivity of 71.4% and a specificity of 80%. Patients with decreased DLCO (n = 18) were not different in age (51.1 more or less than 13.5 vs. 53.5 more or less than 9.3 y, p = 0.5182), sex (male 13.6%, p = 0.6088), prevalence of Raynaud (86.6% vs. 85%, p = 0.6272), sicca syndrome (6.2% vs. 10.5% p = 1.0000) diffuse cutaneous involvement (94.1% vs. 83.3%, p = 0.6026) or esophageal dilatation. The duration of symptoms since diagnosis was no different. Prevalence of pulmonary hypertension assessed by Doppler echocardiography or abnormal nailfold capillaroscopic findings were identical in both populations. Patients with low DLCO had a significatly higher prevalence of anti topoisomerase antibodies. (5/9 vs. 0/11, p = 0.0081) and restrictive lung disease. Patients with low DLCO showed a significantly higher prevalence of abnormal HRCT findings suggestive of ILD (82.3% vs. 5.8%, p less than or equal to 0.0001). We conclude that a low DLCO is a frequent finding in SSc patients, strongly associated with HRCT signs of ILD. We have not found clinical factors predictive for a low DLCO.

Existence of heme oxygenase-carbon monoxide-cyclic guanosine monophosphate pathway in human trabecular meshwork cells in vitro / 华中科技大学学报（医学）（英德文版）

To confirm the existence of heme oxygenase (HO)- carbon monoxide (CO)- cyclic guanosine monophosphate (cGMP) pathway in the cultured human trabecular meshwork cells (HT-MCs) in vitro, and to evaluate the inductive role of hemin on this pathway, HTMCs of the third to fourth generation were cultured in vitro. Reverse transcripase-polymerase chain reaction (RT-PCR) was employed for detection of HO-1 and HO-2 mRNA. Immunohistochemical staining was used to detect HO-1 and HO-2 proteins. Hemin was added into the culture solution. The HO-1 mRNA levels were quantified by RT-PCR. The relative amount of carbon monoxide released into the media was measured with the quantifying carbon monoxide hemoglobin (HbCO) by spectrophotometry. Radioimmunoassay was used to determine changes of cGMP in HTMCs. The results showed that cultured cells had the specific characteristics of HTMCs. Both HO-1 and HO-2 genes were expressed in HTMCs, as well as HO-1 and HO-2 proteins in HTMCs. Hemin induced HO-1 mRNA, HbCO and cGMP in a dose-dependent manner. In conclusion, HO-CO-cGMP pathway exists in the cultured HTMCs and can be induced by hemin. Pharmacological stimulation of HO-CO-cGMP pathway may constitute a novel therapeutic approach to rescuing glaucoma.

The role of endogenous carbon monoxide in the hypoxic vascular remodeling of rat model of hypoxic pulmonary hypertension / 华中科技大学学报（医学）（英德文版）

We investigated the expression of heme oxygenase-1 (HO-1) gene and production of endogenous carbon monoxide (CO) in the rat lung tissue at different time points of chronic hypoxic pulmonary hypertension and the effect of hemin on the expression of HO-1 gene and pulmonary hypertension. A rat model of hypoxic pulmonary hypertension was recreated by exposure to intermittent normobaric hypoxic environment (10% O2). Reverse transcriptase polymerase chain reaction (RT-PCR) was performed to determine the level of HO-1 mRNA in the rat lung tissue and double wave length spectrophotometry was used to evaluate the quantity of COHb in arterial blood. Cardiac catheterization was employed to measure the right ventricular systolic pressure (RVSP) and HE staining was performed in dissected lung tissue to observe the pathological changes of the intra-acinar pulmonary arteries (IAPA). It was found that (1) There was a low level of HO-1 mRNA in normal rat lung tissue, but the level of HO-1 mRNA increased by 2-4 times in the lung tissue of hypoxic rats (P<0.01). The quantity of COHb was 2-3 times those of control group (P<0.01 or P<0.05). These were accompanied by the increased of RVSP and the thickened IAPA; (2) Hemin could keep the HO-1 mRNA and COHb in the hypoxic rat lung tissue at a high level, and partially suppressed the increase of rat RVSP, thereby ameliorating the pathological changes of IAPA. In conclusion, the upregulation of the expression of HO-1 gene and production of CO in the rat lung of hypoxic pulmonary hypertension plays a role of inhibition in the development of hypoxic pulmonary hypertension. Hemin has a therapeutic effect on hypoxic pulmonary hypertension.

Role of crosslinked protein in lung injury following total body irradiation and bone marrow transplantation

The aberrant protein crosslinks formation during lung injury as results total body irradiation (TBI) and bone marrow transplantation (BMT) therapy has been examined as apossible contributory factor in organ or tissue pathogenesis. Female C3HeB/ FeJ mice were used for an experimental animal. Carbon monoxide uptake (V(CO)) was measured at 1, 2, 3, 4 and 5 months after TBI at respective doses of 12, 14, 16 and 18 Gy 16 h prior to syngeneic BMT. Also as a measure of aberrant protein crosslinking in the inured tissues, transglutaminase (TGase)-activities and crosslinked protein were examined along with thrombin, a protease known to activate TGases. Reductions of VCO were detected following TBI and BMT. Activities of thrombin and TGase 1, and crosslinked protein in bronchoalveolar lavage (BAL) fluid of the mice 1 wk after TBI at 12 Gy and BMT were identified and found to be elevated in the treated animals. These findings suggest that elevated levels of crosslinked proteins and TGase I in the bronchoalveolar larvage during the lung injury could have enhanced the organ pathogenesis following TBI and BMT.

Induction of the expression of heme oxygenase gene in PC12 cells by hypoxia / 华中科技大学学报（医学）（英德文版）

To investigate the expression of the HO-1 gene in PC12 cells in hypoxic environment and gain further insight to the role of HO-1 in cerebral ischemia, PC12 cells were exposed to hypoxia environment (95% N2, 5% CO2) for 0.5 h, 1 h, 4 h, 8 h, 12 h, 24 h respectively. The level of HO-1 mRNA was examined by reverse transcriptase polymerase chain reaction (RT-PCR); the volume of COHb in the media were measured spectrophotometrically and the cGMP concentration of PC12 cell extracts was determined by radioimmunoassay. We found that after exposure to hypoxia for 1 h, 4 h, 8 h, 12 h, 24 h, HO-1 mRNA increased by 3%, 4%, 17%, 31% 36% as compared with that in control group respectively (P < 0.01 or P < 0.05); the COHb increased by 12%, 29%, 59%, 88%, 94% as compared with that in control group respectively (P < 0.01 or P < 0.05), and the cGMP concentration were 2.2, 3.4, 5.2, 8.1, 10.9-fold as that of the control group (P < 0.01). We are led to conclude that hypoxia induced the expression of HO-1 gene, the production of endogenous CO, and the concentration of cGMP was elevated as well.

Carbon monoxide: from toxin to endogenous modulator of cardiovascular functions

Carbon monoxide (CO) is a pollutant commonly recognized for its toxicological attributes, including CNS and cardiovascular effects. But CO is also formed endogenously in mammalian tissues. Endogenously formed CO normally arises from heme degradation in a reaction catalyzed by heme oxygenase. While inhibitors of endogenous CO production can raise arterial pressure, heme loading can enhance CO production and lead to vasodepression. Both central and peripheral tissues possess heme oxygenases and generate CO from heme, but the inability of heme substrate to cross the blood brain barrier suggests the CNS heme-heme oxygenase-CO system may be independent of the periphery. In the CNS, CO apparently acts in the nucleus tractus solitarii (NTS) promoting changes in glutamatergic neurotransmission and lowering blood pressure. At the periphery, the heme-heme oxygenase-CO system can affect cardiovascular functions in a two-fold manner; specifically: 1) heme-derived CO generated within vascular smooth muscle (VSM) can promote vasodilation, but 2) its actions on the endothelium apparently can promote vasoconstriction. Thus, it seems reasonable that the CNS-, VSM- and endothelial-dependent actions of the heme-heme oxygenase-CO system may all affect cardiac output and vascular resistance, and subsequently blood pressure

Comparison of single breath and steady state methods for measurement of lung transfer factor for CO in normal subjects and smokers.

Pulmonary diffusing capacity (lung transfer factor for carbon monoxide) has been assessed by the single breath (TLCOSB) and steady state (TLCOSS) techniques in well matched 90 females, 31 non-smoker and 29 smoker males, 18-50 years of age. Both TLCOSB and TLCOSS are significantly lower in females compared to non-smoker males (P < 0.001). Tobacco smoking statistically significantly reduces TLCOSB as well as TLCOSS in smokers as compared to non-smokers. There is a statistically significant correlation of age with TLCOSS and TLCOSB in all the three groups (r = -0.702, -0.360 and 0.300 for TLCOSB and r = -0.481, -0.355 and 0.380 for TLCOSS in non-smoker males, smoker males and females respectively). TLCOSB is 30.43 +/- 4.89, 27.29 +/- 4.54 and 26.13 +/- 3.60 ml/mmHg/min, while TLCOSS is 19.47 +/- 5.26, 16.69 +/- 3.27 and 18.24 +/- 3.78 ml/mm/Hg/min in non-smoker males, smoker males and females respectively. A fairly good correlation between the TLCOSB and TLCOSS in male, both non-smoker and smoker, as well as the female subjects was observed. TLCOSS is lower than TLCOSB in all the three groups. Even in smokers of moderate intensity both of these tests are influenced to a nearly similar extent.

Transfer lung factor for Co (TLCO) and its components (DM & VC) in flour mill workers.

Tlco, Kco (Tlco/Va), Va, Vc and Dm were measured in 25 flour mill workers (non-smoker 9, smoker 16) and 50 healthy control subjects (25 non-smoker, 25 smoker) of similar anthropometric parameters and socio-economic status. Tlco (P < 0.05) was decreased significantly and rest of the parameters were decreased insignificantly in flour mill workers as compared to control subjects. In non-smoker flour mill workers none of the parameters altered significantly as compared to non-smoker control subjects. Statistically significant reduction was seen in smoker workers only in Tlco (P < 0.05) and its component Dm (P < 0.05) as compared to smoker control subjects. Further it has been observed that less duration of flour dust exposure (< 5 years) in all flour mill workers and smoker flour mill workers caused negligible fall in Tlco, whereas longer duration of flour dust exposure (> 5 years) in both the groups caused highly significant fall in Tlco (P < 0.01) which was contributed to by highly significant fall in Dm (P < 0.001).