Association between the CYP1A1 gene polymorphism and susceptibility to emphysema and lung cancer.

Aim-To investigate cytochrome P4501A1 (CYP1A1) polymorphism and susceptibility to emphysema and lung cancer.Methods-A novel polymerase chain reaction (PCR) for genotyping the CYP1A1 polymorphism, corresponding to putative low or high enzyme activity, was developed to genotype lung cancer resection samples which had been assessed macroscopically for the presence of centriacinar and panacinar emphysema. Samples were collected and genotyped from a group of patients with chronic obstructive airways disease. A control group of anonymous blood donations was genotyped to determine the basal levels of the polymorphism in the Scottish population.Results-The high activity allele of the CYP1A1 gene is associated with susceptibility to centriacinar emphysema and lung cancer but not panacinar emphysema. CYP1A1 polymorphism is not linked to lung cancer in the absence of emphysema, nor to chronic obstructive airways disease which is the clinical manifestation of emphysema, particularly of the panacinar type.Conclusions-Susceptibility to emphysema and lung cancer is associated with polymorphism of the P4501A1 gene. A trend towards damage of centriacinar pattern has been detected, which supports the theory that centriacinar emphysema results from local, direct damage to the respiratory bronchioles from exposure to cigarette smoke.

Neutrophil sequestration in lungs removed at surgery. The effect of microscopic emphysema.

Neutrophils within the lungs are considered to play an important role in the pathogenesis of pulmonary emphysema. We have studied the intravascular distribution of reinjected autologous 111In-labeled neutrophils in lung specimens resected 10 min after reinjection from 10 patients undergoing surgery for peripheral bronchogenic tumors. An excess of neutrophils relative to that expected for the 99mTc-labeled erythrocyte blood volume was confirmed in all specimens (range, 3- to 136-fold). In seven specimens which were completely examined, this excess displayed a skewed distribution, with a median neutrophil sequestration of 20-fold excess, and correlated with local blood volume (r = -0.51; p < 0.001). There was also a significant correlation between alveolar wall surface area per unit volume of lung (AWUV) and neutrophil excess, when randomly selected tissue blocks from each specimen were analyzed (r = 0.34, n = 51, p = 0.012). This same trend was demonstrated when whole specimen median values were considered (r = 0.64, n = 7, p = 0.07). Thus in areas of the lungs with lower AWUV values (increasing microscopic emphysema), fewer neutrophils were present. These studies add further support to the view that emphysema per se is not associated with an increased sequestration of pulmonary intravascular neutrophils.

Collagen content of alveolar wall tissue in emphysematous and non-emphysematous lungs.

BACKGROUND: Emphysema is currently defined as "a condition of the lung characterised by abnormal, permanent enlargement of the airspaces distal to the terminal bronchiole, accompanied by destruction of their walls, and without obvious fibrosis." The functional and morphological changes that occur in emphysema have largely been attributed to changes in alveolar elastin rather than in collagen. A study was performed to determine whether the amount of collagen in the alveolar wall changes with age in the lungs of non-smokers and of smokers with different types of macroscopically defined emphysema in relation to a microscopic measurement of lung structure. METHODS: Total alveolar wall collagen was measured (as hydroxyproline) in known volumes of distended lung tissue (by reverse phase high pressure liquid chromatography) in the lungs of non-smokers (n = 23) and in regions sampled away from emphysematous lesions in the lungs of 36 smokers (four with no emphysema, 13 with centriacinar emphysema (CAE), nine with panacinar emphysema (PAE), and 10 with a mixture (MIX) of both PAE and CAE). Mean lung airspace wall surface area per unit volume (AWUV) was calculated from at least six random blocks per lung and on histological sections immediately adjacent to those prepared for collagen measurement with a rapid scanning device (fast interval processor). RESULTS: In non-smokers there was no significant correlation between the amount of collagen in the alveolar wall tissue and either mean lung AWUV or increasing patient age when amounts of collagen were expressed either per unit volume of distended lung (40 mm3 sample) or per unit surface area of airspace wall tissue. Smokers without emphysema had similar amounts of collagen to non-smokers. Lungs with PAE and MIX, but not CAE alone, contained significantly more collagen than normal when expressed per unit volume of airspace wall tissue whereas all groups, including CAE, contained significantly raised amounts of collagen when expressed per unit surface area. CONCLUSIONS: There is no significant age related change in the collagen content of the lungs of non-smokers which suggests that, as AWUV is lost with age, the main collagenous framework is maintained. However, in smokers with emphysema there is a loss of airspace wall tissue in regions remote from the macroscopic lesions that is accompanied by a net increase in collagen mass. The greater accumulation of collagen in MIX lungs than in CAE lungs suggests a greater degree of structural damage, indicative of an alternative pathogenetic mechanism operating between the different types of emphysema. Our results suggest an active alveolar wall fibrosis in emphysema as a consequence of cigarette smoking. It is suggested that the definition of emphysema may require further revision to include such change.

Relation between distal airspace size, bronchiolar attachments, and lung function.

BACKGROUND--Smoking related fixed airway obstruction may be due to airway scarring and narrowing or decreased support due to loss of adjacent alveolar walls. In this study of resected specimens, preoperative pulmonary function was compared with results of a morphometric study of lung structure. METHODS--Morphometric measurements were made on 42 inflation fixed lung specimens as follows: airspace wall surface area per unit volume (AWUV) was measured on at least 25 l mm2 histological fields from each specimen, expressed as a mean, and the mean of the lowest five measurements for each case (LF5). Minimum diameter, maximum diameter, diameter ratio (ellipticality), lumen area, and lumen circumference were measured on at least 16 non-respiratory bronchioles from each lung. Peribronchiolar alveolar support was measured as mean interalveolar attachment distance (IAAD). Measurements of pulmonary function included forced expiratory volume in one second (FEV1) (absolute and % predicted values; n = 42), slope of phase III (single breath nitrogen test; n = 28), closing volume (expressed as a percentage of vital capacity (CV/VC%); n = 28). RESULTS--Bronchiolar size was not independently related to the tests of lung function used. Percentage predicted FEV1 was related to mean IAAD, ellipticality, and mean AWUV. CV/VC% showed significant relation with ellipticality, mean AWUV, and LF5 AWUV. Slope of phase III increased with increasing IAAD. Significant correlations were found between ellipticality and AWUV (mean and LF5), and between ellipticality and IAAD. Both IAAD and ellipticality were significantly increased in patients with abnormally low FEV1. CONCLUSION--Destruction of airspace walls, particularly those attached to the peripheral bronchioles, is more influential in determining airflow limitation than bronchiolar size.

Age related increase in the intra-alveolar macrophage population of non-smokers.

The alveolar macrophage population is higher in smokers than in non-smokers. An age related increase in the alveolar macrophage number in the lungs of non-smokers has been found, whether expressed per unit lung volume or per unit lung surface area.

Microscopic emphysema in relation to age and smoking habit.

BACKGROUND: The increases in airspace size within the human lung associated with microscopic emphysema can be assessed by measuring the airspace wall surface area per unit volume of lung tissue (AWUV). In a previous study the limits of normality of AWUV with age were estimated in lifelong non-smokers by the 95% prediction limits of the regression line for these variables. The aims of this study were to study the incidence of microscopically assessed emphysema in a group of smokers and to examine the influence of smoking habit on the susceptibility to and severity of microscopically assessed emphysema. METHODS: AWUV was measured on tissue sections from 125 lung specimens obtained from tobacco smokers (mean age 61.1 (range 33-85) years) with the fast interval processor, a rapid automatic scanning device. The mean AWUV value was calculated for each specimen and this figure was plotted against the age of the subject. The limits of normal AWUV were plotted, and AWUV values below these limits were taken as indicative of microscopically assessed emphysema. Details of the number of cigarettes smoked each day were obtained for 97 of the smokers. These subjects were grouped according to smoking habit: group 1, 1-19 cigarettes/day; group 2, 20-29 cigarettes/day; group 3, at least 30 cigarettes/day. The AWUV results from each of these groups were then assessed. RESULTS: Mean AWUV decreased with age in this group of smokers, but only 26% had microscopically assessed emphysema, indicating that within the group there were two subgroups of smokers with differing susceptibility to microscopically assessed emphysema. There were no sex differences in the incidence of microscopically assessed emphysema, nor were the incidence and severity increased with increased daily cigarette consumption. CONCLUSIONS: Susceptibility to microscopically assessed emphysema was found to be similar in male and female smokers. Daily cigarette consumption did not appear to be the primary factor influencing the susceptibility to or severity of microscopically assessed emphysema. Susceptibility differences within the smoking population should be taken into consideration in studies of the pathogenesis of emphysema.

Airspace size in lungs of lifelong non-smokers: effect of age and sex.

BACKGROUND: Emphysema is defined as the abnormal enlargement of airspaces distal to the terminal bronchiole, but the limits of normality of airspace size have never been defined. The aims of the study were to examine the effects of age and sex on airspace size in non-smokers and to define the limits of normal airspace size. METHODS: Airspace size was measured in terms of airspace wall surface area per unit volume of lung tissue (AWUV). AWUV was measured on histological sections of lung tissue with an automated scanning system, the fast interval processor. Thirty eight lifelong non-smokers were studied, 15 male and 23 female, with an age range of 21-93 years. Macroscopic emphysema was assessed semi-quantitatively on the midsagittal slice of each lung specimen. RESULTS: The relation between mean AWUV and age was negative (r = -0.78). This relation was linear, and the 95% prediction limits of its regression line were used as the limits of normality of AWUV in this sample. The AWUV-age relationship was similar in the men and women studied. Localised areas of macroscopic emphysema were found in three specimens whose mean AWUV measurements were within the normal range. CONCLUSIONS: There is a normal increase in airspace size associated with advancing age in adult lungs. The limits of normal AWUV between ages 21 and 93 years have been defined, and it is proposed that lungs with a mean AWUV below the 95% prediction limit should be considered as having emphysema.

Intra-alveolar macrophage numbers in current smokers and non-smokers: a morphometric study of tissue sections.

BACKGROUND: The alveolar macrophage is believed to be important in the defence of the lung and possibly in the pathogenesis of lung disease. Cell counts in bronchoalveolar lavage fluid have suggested that smokers have an increased number of alveolar macrophages but have not enabled the number to be related to a measure of lung structure. METHODS: The number of alveolar macrophages was counted in histological sections from lung resection specimens from a group of smokers and non-smokers. The results were related to a measurement of lung structure obtained by means of an automated morphometric technique and expressed in terms of units of lung volume or of lung surface area. RESULTS: The smokers had a significantly increased number of alveolar macrophages per unit lung volume and per unit surface area, through the relative increase was less than has appeared from bronchoalveolar lavage studies. When smokers and non-smokers with similar lung structure were compared the smokers had more alveolar macrophages, indicating that smoking and not loss of lung structure is responsible for the increase. CONCLUSIONS: Smokers had more alveolar macrophages than non-smokers when the number was expressed quantitatively with respect to the underlying architecture. Changes in cell populations postulated to be important in the pathogenesis of disease within the lung should be related to lung architecture because this may vary considerably between individuals.

Changes in neutrophil morphology and morphometry following exposure to cigarette smoke.

Acute cigarette smoking delays neutrophils within the pulmonary circulation in some smokers. Evidence from an in-vitro Micropore filter model of the pulmonary capillaries indicates that this may be due to a smoke induced decrease in cell deformability. In order to determine whether changes in cell shape are associated with the observed decrease in neutrophil deformability following smoke exposure, cell morphology, using scanning electron microscopy, and morphometric measurements, made using transmission electron microscopy, were performed on aliquots of neutrophils harvested from whole blood in non-smoking subjects before and after exposure in vitro to cigarette smoke. Smoke exposure increased the maximum diameter and circumference of neutrophils, without changing their area. There was also a change in the maximum to minimum cell diameter ratio, which indicated that the cells had become less spherical. Scanning electron microscopy showed that smoke exposed cells had developed blebbing of their surface membranes, suggestive of an oxidative injury to the cell membrane rather than the shape changes associated with cell activation. These changes in the morphology and morphometry of smoke exposed neutrophils may contribute to the reduction in cell deformability induced by cigarette smoke.

Microscopic and macroscopic measurements of emphysema: relation to carbon monoxide gas transfer.

BACKGROUND: Studies of the relation between the severity of structural change in emphysema and physiological abnormality have been based on macroscopic assessments, which have not been truly quantitative or sensitive enough to detect early changes. With a highly reproducible method for measuring emphysema using histological sections and a semiautomatic image analysis system, this quantitative assessment of emphysema was compared with a semiquantitative macroscopic assessment of emphysema and measurements of carbon monoxide gas transfer. METHODS: Microscopic and macroscopic measurements of emphysema on 44 thoracotomy specimens were compared; only two were from non-smokers. Airspace wall surface area per unit volume was measured microscopically with an automatic image analyser and expressed as both the mean airspace wall surface area per unit volume and the mean value of the five fields with the lowest values. Macroscopic emphysema was measured directly on a tracing of the midsagittal slice using a digitising tablet attached to a microcomputer and expressed as a percentage of the total area of lung. In cases with centriacinar emphysema the number of discrete lesions was counted. RESULTS: The area of macroscopic emphysema ranged from 0 to 78% of the total area of lung examined, but most patients had less than 1% involvement so that the distribution was highly skewed. Both mean airspace wall surface area per unit volume and the mean of five fields with the lowest airspace wall surface area per unit volume were normally distributed, with mean airspace areas ranging from 8.8 to 25.4 mm2/mm3 (mean 18.1 mm2/mm3). In lobes with centriacinar emphysema the number of discrete lesions correlated with airspace wall surface area per unit volume and with preoperative carbon monoxide transfer factor (TLCO) per unit lung volume. However, other measurements of macroscopic emphysema did not correlate with loss of alveolar wall surface area, and there was considerable overlap between subjects with no or minimal macroscopic emphysema and those with more severe disease. TLCO correlated with both mean airspace wall surface area per unit volume and the mean of five fields with the lowest airspace wall surface area per unit volume but not with the severity of macroscopic emphysema. CONCLUSION: If emphysema is to be quantified it must be measured microscopically; macroscopic measurements do not, in general, reflect the microscopic loss of airspace wall.

New automated technique for assessing emphysema on histological sections.

The assessment of emphysema in human lungs has traditionally been based on observations made on whole lung slices. These methods are inappropriate for the study of early emphysema, because as much as 75% of the alveolar wall surface area may have been lost by the time airspaces are visible to the naked eye. A new, automated image analysis system, the Fast Interval Processor (FIP), was used to measure airspace wall surface area per unit volume of lung tissue (AWUV). AWUV was measured on histological sections of lung tissue and expressed in mm2/mm3. The study sample consisted of resection specimens from 40 patients (32 men and 8 women whose ages ranged from 23-74 years). Histological sections from the inflated specimens were scanned using the FIP, and a mean AWUV value was calculated for each. The intra- and interobserver reproducibility of this method of measuring AWUV were examined. The results obtained using the FIP were also compared with those from an established image analysis system. The FIP is a fast, efficient technique which gave highly reproducible results comparable with those obtained with an established and much more time consuming measuring technique.

Fortification of curry powder with NaFe(111)EDTA in an iron-deficient population: initial survey of iron status.

A survey of iron status was conducted in 984 volunteers (404 males and 580 females) from an Fe-deficient population before an Fe-fortification trial. Hemoglobin, percentage saturation of transferrin, and serum ferritin were used to assess Fe status and to calculate body Fe stores. Almost 30% of males and 60% of females had evidence of Fe deficiency. The distribution of body Fe stores for both males and females was shifted to the left compared with a population in the United States. In females 24% had Fe-deficiency anemia, 13% Fe-deficient erythropoiesis, and 16% depleted stores. Multiple regression analysis failed to show any relationship in women between age, parity, and duration of menses and measurements of Fe status. In males Fe deficiency was more frequent for those less than 18 y and alcohol abusers had increased serum ferritin and calculated body Fe compared with nondrinkers.

Fortification of curry powder with NaFe(111)EDTA in an iron-deficient population: report of a controlled iron-fortification trial.

A targeted, double-blind controlled iron fortification trial using Fe(111)-EDTA in masala (curry powder) was directed towards an Fe-deficient Indian population for 2 y. The Fe status of the fortified group improved more than that of control subjects. Improvement reached significance over control subjects for females in hemoglobin (p = 0.0005), ferritin (p = 0.0002), and body Fe stores (p = 0.001) and for males in ferritin (p = 0.04). The prevalence of Fe-deficiency anemia (IDA) decreased from 22 to 5% in fortified females. Premenopausal women, multipara women, and women with prolonged menstruation or initial IDA benefitted most from fortification. The mean increase in body Fe stores in females with initial IDA was 9.0 +/- 1.3 mmol, representing an increased absorption of 12 mumol/d. Fortified subjects with normal Fe status did not accumulate excessive body Fe and there was no alteration in serum Zn concentrations. Targeted fortification is a safe and effective means of combatting Fe deficiency.

The effects of fruit juices and fruits on the absorption of iron from a rice meal.

The effects of the chemical composition of fruit juices and fruit on the absorption of iron from a rice (Oryza sativa) meal were measured in 234 parous Indian women, using the erythrocyte utilization of radioactive Fe method. The corrected geometric mean Fe absorptions with different juices varied between 0.040 and 0.129, with the variation correlating closely with the ascorbic acid contents of the juices (rs 0.838, P less than 0.01). Ascorbic acid was not the only organic acid responsible for the promoting effects of citrus fruit juices on Fe absorption. Fe absorption from laboratory 'orange juice' (100 ml water, 33 mg ascorbic acid and 750 mg citric acid) was significantly better than that from 100 ml water and 33 mg ascorbic acid alone (0.097 and 0.059 respectively), while Fe absorption from 100 ml orange juice (28 mg ascorbic acid) was better than that from 100 ml water containing the same amount of ascorbic acid (0.139 and 0.098 respectively). Finally, Fe absorption from laboratory 'lemon juice' (100 ml orange juice and 4 g citric acid) was significantly better than that from 100 ml orange juice (0.226 and 0.166 respectively). The corrected geometric mean Fe absorption from the rice meal was 0.025. Several fruits had little or no effect on Fe absorption from the meal (0.013-0.024). These included grape (Vitis vinifera), peach (Prunus persica), apple (Malus sylvestris) and avocado pear (Persea americana). Fruit with a mild to moderate enhancing effect on Fe absorption (0.031-0.088) included strawberry (Fragaria sp.) (uncorrected values), plum (Prunus domestica), rhubarb (Rheum rhaponticum), banana (Musa cavendishii), mango (Mangifera indica), pear (Pyrus communis), cantaloup (Cucumis melo) and pineapple (Ananas comosus) (uncorrected values). Guava (Psidium guajava) and pawpaw (Carica papaya) markedly increased Fe absorption (0.126-0.293). There was a close correlation between Fe absorption and the ascorbic acid content of the fruits tested (rs 0.738, P less than 0.0001). There was also a weaker but significant correlation with the citric acid content (rs 0.55, P less than 0.03). Although this may have reflected a direct effect of citric acid on Fe absorption, it should be noted that fruits containing citric acid also contained ascorbic acid (rs 0.70, P less than 0.002).(ABSTRACT TRUNCATED AT 400 WORDS)

Factors influencing the absorption of iron from soya-bean protein products.

The absorption of iron from soya-bean (Glycine hispida)-based and milk-based infant formulas was assessed in 138 multiparous Indian women, using the erythrocyte utilization of radioactive Fe method. Fe absorption was significantly greater from the basal milk formula (1.5 g protein) than it was from the basal soya-bean formula (2.3 g protein), with geometric mean values of 0.083 and 0.044 respectively. Ascorbic acid markedly increased Fe absorption from the milk-based formula in a dose-dependent fashion. The increase was fivefold when the ascorbic acid:Fe ratio on a weight-for-weight basis was 6:1 and over tenfold when it was 20:1. In contrast, ascorbic acid had a less-marked effect on the absorption of Fe from the soya-bean-based formula, with only a two- to threefold increase at an ascorbic acid:Fe ratio of 20:1. The geometric mean Fe absorption from the soya-bean formula (1.27 mg Fe, 2.3 g isolated soya-bean protein (ISP] was somewhat less than that from the same amounts of ISP and ascorbic acid made up in milk (0.075 and 0.113 respectively). However, a direct comparison between the soya-bean formula in milk and in water showed no significant difference (0.043 and 0.060 respectively). Fe absorption from a drink containing 10 g ISP and 30 mg ascorbic acid was significantly better than that from a similar drink containing the soya-bean flour from which ISP is extracted (0.044 and 0.027 respectively). Heating ISP to 200 degrees for 2 h before its use had no effect on Fe availability.(ABSTRACT TRUNCATED AT 250 WORDS)

The relative effect of ascorbic acid on iron absorption from soy-based and milk-based infant formulas.

The effect of varying concentrations of ascorbic acid on the absorption of iron from a soy-based infant milk formula containing 6 mg iron/100 g was examined in 64 adult Indian females using the extrinsic radioactive tag method. The corrected geometric mean absorption from the basic soy formula was only 1.8%. Addition of ascorbic acid in a concentration of 40 mg/100 g, did not significantly increase absorption (3.3%; t = 1.8, p greater than 0.07) but raising the concentration to 80 mg/100 g did so (6.9%; t = 2.4, p less than 0.02). No further significant increase was noted when the concentration of ascorbic acid was increased to 160 mg/100 g (7.7%; t = 0.4, p greater than 0.7). The inhibitory effect of soy on iron absorption was further demonstrated by a direct comparison between the soy-based formula and a similar product based on cows' milk. The comparison was made at two concentrations of ascorbic acid. At 40 mg/100 g the geometric mean iron absorption from the soy formula was 2.4% compared with 5.3% from the milk formula (t = 2.8, p less than 0.02), while the corresponding values at 80 mg ascorbic acid/100 g were 7.2 and 19.5%, respectively (t = 3.4, p less than 0.02). The present results confirm the marked inhibitory effect of soy protein on iron absorption and calculations from the absorption figures suggest that such formulas should contain at least 12 mg/100 g iron together with ascorbic acid in a molar ratio of approximately 4:1 if they are to be adequate in terms of iron nutrition.

Factors affecting the absorption of iron from cereals.

Non-haem-iron absorption from a variety of cereal and fibre meals was measured in parous Indian women, using the erythrocyte utilization of radioactive Fe method. The present study was undertaken to establish whether alteration of the phytate and polyphenol contents of sorghum (Sorghum vulgare) affected Fe absorption from sorghum meals, and to assess the influence of fibre on Fe absorption. Removing the outer layers of sorghum grain by pearling reduced the polyphenol and phytate contents by 96 and 92% respectively. This treatment significantly increased the geometric mean Fe absorption from 0.017 to 0.035 (t 3.9, P less than 0.005). The geometric mean Fe absorption from a sorghum cultivar that lacked polyphenols (albino sorghum) was 0.043, which was significantly greater than the 0.019 absorbed from bird-proof sorghum, a cultivar with a high polyphenol content (t 2.83, P less than 0.05). Fe was less well absorbed from the phytate-rich pearlings of the albino sorghum than from the pearled albino sorghum (0.015 v. 0.035 (t 8.4, P less than 0.0005]. Addition of sodium phytate to a highly Fe-bioavailable broccoli (Brassica oleracea) meal reduced Fe absorption from 0.185 to 0.037. The geometric mean Fe absorption from malted sorghum porridge was 0.024 when 9.5 mg ascorbic acid were added and 0.094 when the ascorbic acid was increased to 50 mg (t 3.33, P less than 0.005). This enhancing effect of 50 mg ascorbic acid was significantly depressed to 0.04 by tea (t 38.1, P less than 0.0005).(ABSTRACT TRUNCATED AT 250 WORDS)