Spreading rate dependence of gravity anomalies along oceanic transform faults.

Mid-ocean ridge morphology and crustal accretion are known to depend on the spreading rate of the ridge. Slow-spreading mid-ocean-ridge segments exhibit significant crustal thinning towards transform and non-transform offsets, which is thought to arise from a three-dimensional process of buoyant mantle upwelling and melt migration focused beneath the centres of ridge segments. In contrast, fast-spreading mid-ocean ridges are characterized by smaller, segment-scale variations in crustal thickness, which reflect more uniform mantle upwelling beneath the ridge axis. Here we present a systematic study of the residual mantle Bouguer gravity anomaly of 19 oceanic transform faults that reveals a strong correlation between gravity signature and spreading rate. Previous studies have shown that slow-slipping transform faults are marked by more positive gravity anomalies than their adjacent ridge segments, but our analysis reveals that intermediate and fast-slipping transform faults exhibit more negative gravity anomalies than their adjacent ridge segments. This finding indicates that there is a mass deficit at intermediate- and fast-slipping transform faults, which could reflect increased rock porosity, serpentinization of mantle peridotite, and/or crustal thickening. The most negative anomalies correspond to topographic highs flanking the transform faults, rather than to transform troughs (where deformation is probably focused and porosity and alteration are expected to be greatest), indicating that crustal thickening could be an important contributor to the negative gravity anomalies observed. This finding in turn suggests that three-dimensional magma accretion may occur near intermediate- and fast-slipping transform faults.

The effect of transforming growth factor beta on rates of procollagen synthesis and degradation in vitro.

Transforming growth factor beta (TGF beta) is known to stimulate procollagen production and steady-state levels of procollagen mRNAs, but its ability to affect post-translational processing of procollagen has been little studied. This paper demonstrates the application of recently developed ultrasensitive methods for measuring hydroxyproline to assess rates of procollagen synthesis and degradation in vitro with and without TGF beta. Foetal rat fibroblasts synthesized 8.63 +/- 0.21 pmol hydroxyproline/micrograms DNA per h, which corresponds to approx. 40 molecules of procollagen/cell per s. Addition of TGF beta to cultures increased total amounts of procollagen synthesized and degraded by 112% and 82%, respectively, but there was a significant decrease in the proportion of procollagen degraded (control, 38.0 +/- 1.1%; TGF beta, 32.3 +/- 0.9%; P less than 0.005). This study demonstrates a novel mechanism which may contribute to the TGF beta-induced increase in procollagen production by fibroblasts.

Increased collagen synthesis and decreased collagen degradation in right ventricular hypertrophy induced by pressure overload.

OBJECTIVE: The aim was to examine the effect of pressure overload in rabbits on ventricular collagen metabolism and procollagen gene expression. METHODS: Right ventricular hypertrophy was induced by banding the pulmonary artery such that the diameter of the vessel was reduced by 50%, and animals killed in groups after two and 14 days. Collagen synthesis and degradation of newly synthesised collagen were assessed following a single intravenous injection of 3H-proline with a flooding dose of non-radioactive proline, given 3 h before the animals were killed. Northern and slot blot analyses were performed to measure procollagen alpha 1(I) mRNA. RESULTS: The fractional collagen synthesis rate increased sixfold in the right ventricle only 2 d after pulmonary artery banding (p < 0.001), then fell to just over double the control value by 14 d (p < 0.05 from control). The proportion of newly synthesised collagen degraded decreased from 50.7(SD 12.8)% to 26.8(15.8)% in 2 d (p < 0.05) and remained at this level. The procollagen alpha 1(I) mRNA level increased by more than fourfold in the right ventricle 2 d after the onset of pressure overload, and was less than three times control levels at 14 d. CONCLUSIONS: The development of right ventricular hypertrophy is associated with a rapid increase in collagen production, with regulation at multiple sites in the biosynthetic pathway. This regulation occurs at both transcriptional and post translational levels.

Adhesive interactions between fibroblasts and polymorphonuclear neutrophils in vitro.

Although the in vivo interaction between polymorphonuclear neutrophils (PMN) and fibroblasts may be important, these pathways have not been well studied. We have investigated the adherence of PMN to monolayers of human fetal lung fibroblasts, using a microtiter plate assay based upon the uptake by cells of the vital stain Rose Bengal. Stimulation with phorbol myristate acetate (PMA) caused a significant increase of adherence over basal levels which was rapid in onset and plateaued at 5 min. Adhesion was dependent on the leucocyte integrin family of glycoproteins, notably on Mac-1, since monoclonal antibodies toward the beta chain (CD18) and alpha chain (CD11b) of Mac-1 almost completely suppressed PMA-induced PMN adhesion (88% and 77% inhibition, respectively). Adhesion was also inhibited by the peptides RGDS and GRGDS (24.2% and 26.6%, respectively using 1 mM peptide). Prestimulation of fibroblasts for longer time periods (5 and 24 h) with interleukin 1 alpha and tumor necrosis factor alpha, but not transforming growth factor beta, also resulted in a significant increase in adhesion of unstimulated PMN (after 24 h preincubation, 10 U/ml IL1 alpha stimulated adhesion by 179% of control, 500 U/ml TNF alpha by 157%). This indicated that there are both PMN- and fibroblast-dependent pathways for PMN adhesion. Components of the extracellular matrix of fibroblasts do not appear to play important roles in the adhesion process since addition of fibronectin and type IV collagen, or of purified antibodies to fibronectin and types I and IV collagen, did not affect PMA-induced PMN adhesion.(ABSTRACT TRUNCATED AT 250 WORDS)

Thrombin stimulates fibroblast chemotaxis and replication.

The serine protease alpha-thrombin, a product of the circulating zymogen prothrombin, plays multiple roles in homeostasis and coagulation. During blood clotting, it is present within fibrin matrices and is likely to be presented to local cell populations. It is known to be a fibroblast mitogen, but its effects on fibroblast recruitment have not been assessed. Here we compared the effect of human alpha-thrombin on chemotaxis and proliferation of human and rat skin fibroblasts and assessed the mechanism of these actions. Fibroblast chemotaxis was assayed using a 48-well Boyden chamber and replication assessed by a spectrophotometric method, based upon the uptake and subsequent elution of methylene blue by fibroblasts. Two fibroblast cell lines were used; fetal rat skin (FR) and newborn human foreskin (HS68). Human alpha-thrombin stimulated FR fibroblast chemotaxis over a wide range of doses (10(-12) M to 10(-7) M). Maximal migration was seen at 10(-10) M; 39 +/- 2.5 cells/high power field (h.p.f.) compared with 19 +/- 3 cells/h.p.f. for media control. In the same assay platelet-derived growth factor, a well characterized fibroblast chemoattractant, caused a maximal stimulation of 44 +/- 5 cells/h.p.f. at a concentration of 3 x 10(-9) M. A similar stimulation was observed with HS68 fibroblasts, although for this cell line maximal chemotaxis (190 +/- 12.5% of control) was seen at 10(-8) M thrombin. Fibroblast replication was optimal at 1.25 x 10(-9) M thrombin (134 +/- 4 and 127 +/- 5% of control for FR and HS68, respectively).(ABSTRACT TRUNCATED AT 250 WORDS)

Collagen production and replication by cardiac fibroblasts is enhanced in response to diverse classes of growth factors.

The tissue distribution and cellular effects of platelet-derived growth factor (PDGF), basic fibroblast growth factor (bFGF), transforming growth factor beta-1 (TGF beta 1) and insulin-like growth factor 1 (IGF-1) suggest a potential role for these factors in cardiovascular matrix deposition. The objective of this study was to assess the capacity of these growth factors to promote cardiac fibroblast collagen production and replication in vitro which will lead to studies identifying their role in vivo during cardiac development and disease. Fibroblasts were isolated from fetal rat hearts by explant culture, and their response to growth factors was assessed with respect to fibroblast replication and collagen synthesis. Fibroblast replication was stimulated by PDGF and by bFGF.IGF-1 and TGF beta 1 had no effect on fibroblast replication. Collagen production was stimulated by all of the growth factors tested in order of potency TGF beta 1 > PDGF, IGF > bFGF. None of the growth factors affected the proportion of newly synthesized collagen rapidly degraded. We have shown that TGF beta 1, PDGF, bFGF and IGF-1 are all capable of increasing collagen deposition by cardiac fibroblasts by either stimulating fibroblast replication or collagen synthesis or both. The sensitivity of cardiac fibroblasts to these factors is consistent with their playing a role in the rapid changes in cardiac collagen deposition seen during development and disease.

Cytokine regulation of mesothelial cell proliferation in vitro and in vivo.

Previous studies have demonstrated mitogenic effects of several mediators on mesothelial cells in vitro, but their effects in vivo have not been investigated. The aim of this study was to examine the effects of various cytokines on normal mesothelial cell proliferation in vitro and in vivo and correlate the findings in both assay systems. In vitro proliferation was assessed using a technique based on the uptake and subsequent release of methylene blue. Autoradiographic methods were applied in a murine model to assess mitogenic activity of these factors on mesothelium in vivo. In vitro data demonstrated a dose-dependent increase in human mesothelial cell proliferation by all mediators examined: at optimal concentrations, proliferation was enhanced between 26.53 +/- 3.77% standard deviation (SD), p < 0.001 for fibroblast growth factor-2 (FGF-2) and 114.58 +/- 6.97%, p < 0.001 for platelet-derived growth factor-AB (PDGF-AB) above control medium. In vivo, DNA synthesis in mesothelial cells was stimulated by FGF-2 (29.52 +/- 5.85% labeled cells, compared with 7.04 +/- 4.36% for control medium; p < 0.001), tumor necrosis factor-alpha (TNF-alpha; 13.14 +/- 4.55% compared with 7.23 +/- 2.85; p < 0.005) and PDGF-BB (11.53 +/- 4.74% compared with 4.67 +/- 3.48%; p < 0.005). Transforming growth factor-beta1 (TGF-beta1) and epidermal growth factor (EGF) had no effect on DNA synthesis in mesothelial cells in vivo. It is concluded that FGF2, TNF-alpha, and PDGF stimulate mesothelial cell proliferation in vitro and in vivo, whereas TGF-beta1 and EGF only had a mitogenic effect in vitro at the concentrations examined. The mitogenic potency of the different PDGF isoforms in vitro was consistent with PDGF-alpha and beta receptor expression.

Thrombin.

Thrombin is a multifunctional serine protease which plays a central role in haemostasis by regulating platelet aggregation and blood coagulation. It is formed from its precursor prothrombin following tissue injury and converts fibrinogen to fibrin in the final step of the clotting cascade. It also promotes numerous cellular effects including chemotaxis, proliferation, extracellular matrix turnover and release of cytokines. These actions of thrombin on cells have been implicated in tissue repair processes and in the pathogenesis of inflammatory and fibroproliferative disorders such as pulmonary fibrosis and atherosclerosis. Thrombin mediates its cellular effects by proteolytically activating cell surface receptors. Presently, two such receptors have been described and their roles in regulation of these functions are currently being investigated. The discovery of multiple thrombin receptors creates the possibility of selective receptor blockade of specific thrombin mediated events. New drugs with these actions should add to our current repertoire of thrombin inhibitors used to treat thrombotic diseases.

Mechanisms of tissue repair: from wound healing to fibrosis.

To set the scene for this Directed Issue on Mechanisms of Tissue Repair of The International Journal of Biochemistry and Cell Biology, this introductory overview briefly describes the process of wound healing and highlights some of the key recent advances in this field of research. It emphasizes the importance of cell-cell and cell-matrix interactions, particularly relating to the role of cell surface adhesion molecules, and describes developments that have led to a better understanding of the dynamic nature of matrix turnover with reference to negative and positive mediators that regulate procollagen gene expression and protein production. An important component of this Directed Issue is concerned with the development of tissue fibrosis, which accompanies a number of disease states and demonstrates remarkable parallels with the normal wound healing process; excessive amounts of matrix are laid down but the resolution of scarring, which would be anticipated in wound healing, is impaired. The possible mechanisms involved in fibrosis are discussed here. Since cytokines play an important role in regulating cell function such as proliferation, migration and matrix synthesis, it is the balance of these mediators which is likely to play a key role in regulating the initiation, progression and resolution of wounds. Finally, this review highlights areas of tissue repair research in which recent developments have important clinical implications that may lead to novel therapeutic strategies.

The pathogenesis of pulmonary fibrosis: is there a fibrosis gene?

Interstitial fibrosis is seen in the lung in response to a variety of insults, and often appears stereotypical in terms of its clinical and pathological features. However, exposure to a known aetiological factor does not always lead to fibrosis. For example in bleomycin-induced pulmonary fibrosis, a wide variation in response is seen both in humans and in animal models, which is not completely accounted for by known risk factors. These observations and the existence of a number of familial forms of lung fibrosis suggest a genetic predisposition. Current hypotheses concerning the pathogenesis of pulmonary fibrosis propose an initial stage involving the influx of inflammatory cells into the interstitium. These cells, together with activated resident cells are then thought to release polypeptide mediators that stimulate the fibroblast proliferation and matrix protein synthesis typical of these disorders. Genetic influences could have an important role in regulating a number of these events, altering the immunological response to injury or modulating collagen metabolism in the lung. However, despite recent advances in molecular genetic techniques, there have been few human studies to date. Most have concentrated on genetic loci with a high degree of polymorphism such as the human leucocyte antigen (HLA) system and yield conflicting results. Others offer tantalising but as yet, incomplete insights into the mechanisms involved. Defining the genetic abnormalities underlying both the familial forms of pulmonary fibrosis and the variations seen in response to lung injury should enhance our understanding of the pathogenic processes and help to focus research in this area.

Age-related alterations in collagen and total protein metabolism determined in cultured rat dermal fibroblasts: age-related trends parallel those observed in rat skin in vivo.

The cultured fibroblast has been extensively used as a model system to study aging. However, few studies have examined the veracity of observations obtained in cultured fibroblasts aged in vitro to those made in animal tissues in vivo. This paper compares age-related alterations in collagen metabolism measured in cultured cells with previously reported results in the aging rat (Mays et al. (1991) Biochem. J. 276, 307-313). Age-related changes in collagen synthesis in rat skin fibroblasts in vitro over 30 population doublings were determined based on the production of hydroxy-[14C]proline. Degradation of newly synthesized collagen was based on the appearance of free hydroxy-[14C]proline in the culture system. Total protein synthesis rates were based on the incorporation of [14C]proline into proteins. In vitro rates of collagen synthesis decreased 5-fold over 30 population doublings (P < 0.05). Degradation of newly synthesized collagen increased from 33.0 +/- 0.8% (n = 4, SEM) to 45.2 +/- 1.1% (n = 4; P < 0.05) over the same period, with a maximum after 25 population doublings of 55.8 +/- 1.1% (n = 4). Total protein synthesis rates decreased by one-half over 30 population doublings (P < 0.05). The results indicated that collagen production decreased as cells aged in vitro and that this was due to both changes in synthesis and degradation. The results demonstrate that age-related alterations in collagen and total protein metabolism of skin fibroblasts in culture were similar to those reported previously for skin in vivo, suggesting that for studies of these processes, fibroblasts in culture provide an appropriate model.

Changes in collagen metabolism in response to endothelin-1: evidence for fibroblast heterogeneity.

Endothelin-1 (Et-1) is a 21-amino acid peptide primarily synthesized by endothelial cells. It was originally classified as a potent vasoconstrictor but recent evidence suggests that it also possesses a wide variety of non-vascular actions. It stimulates fibroblast and smooth muscle cell proliferation and it has been shown to stimulate fibroblast collagen metabolism. However, studies on its ability to regulate collagen production remain incomplete, and its effect on post-translational processing of procollagen has not been studied. This report details the effect of Et-1 on the rates of procollagen synthesis and degradation in two fibroblast cell lines; human foetal lung (HFL-1) and whole foetal rat fibroblasts (Rat 2). Fibroblast cultures were incubated for 24 hr in the presence or absence of Et-1 before procollagen metabolism was determined by measuring hydroxyproline. Non-collagen metabolism was also determined in these cultures from the uptake of tritiated phenylalanine. Et-1 stimulated procollagen synthesis in HFL-1 fibroblasts and reduced synthesis in Rat 2 cells. The response was dose dependent with the greatest effect at 1.10(-6) M Et-1 for both cell types (155 +/- 6% of control (mean +/- SD, n = 6, P < 0.01) and 61 +/- 4% of control (n = 4, P < 0.01) for HFL-1 and Rat 2 fibroblasts, respectively). Non-collagen protein synthesis was increased to 148 +/- 5% of control (P < 0.05) at 1.10(-6) M Et-1. Non-collagen protein synthesis remained unaffected in the HFL-1 fibroblast cultures. Procollagen degradation, expressed as a proportion of total procollagen synthesis, was decreased in HFL-1 fibroblasts (control, 29 +/- 2%; Et-1, 1.10(-6) M; 21 +/- 2%; P < 0.01), and increased in Rat 2 fibroblasts (control 42 +/- 1%; Et-1, 1.10(-6) M; 49 +/- 1%; P < 0.01). Blocking of the EtA receptor for Et-1, using the receptor antagonist-BQ123, abolished the effect of Et-1 on procollagen metabolism in both cell types. These results suggest that different populations of fibroblasts exhibit heterogeneous responses to Et-1. It is concluded that Et-1 may play an important role in the extent and distribution of fibrosis seen in diseases associated with the overproduction of Et-1.

Age-related changes in rates of protein synthesis and degradation in rat tissues.

It has been hypothesised that a diminished capacity for protein synthesis and degradation underlies a decreased adaptability to environmental stimuli seen during ageing. In this study rates of total protein synthesis and degradation were examined in rats between 1 and 24 months of age. Synthesis rates in heart, lung, skeletal muscle and skin were based on the uptake of [14C]proline into protein when administered with a flooding dose of unlabelled proline. Degradation rates were derived from the difference between protein deposition and synthesis rates. Total protein synthesis rates in 1-month-old animals ranged from 20.4 +/- 1.3% per day (S.E.M.) in skeletal muscle to 39.6 +/- 1.3% per day in lung. In heart, lung and skeletal muscle, synthesis rates decreased 2-fold during the first 6 months of life, while over the same period in skin they decreased 6-fold. Degradation accounted for the bulk of protein synthesised at all ages, and the age-related changes for rates for breakdown closely mirrored those for synthesis. These results, do not support the hypothesis that a general decrease in protein turnover underlies a diminished adaptability in older animals.

Age-related changes in the proportion of types I and III collagen.

Previous studies of the age-related changes in interstitial collagens, have suggested that the proportion of type III collagen compared with type I decreases with age. In this study collagen concentration and the proportion of types were measured in heart, lung and skin of male Lewis rats aged between 1 day and 2 years. Collagen concentration, based on hydroxyproline levels, increased in all tissues up until 6 months of age, thereafter it increased in heart and lung, yet decreased in skin. The relative proportions of types I and III collagen were assessed after cyanogen bromide digestion of the tissue and separation of the resultant peptides by sodium dodecyl sulphate polyacrylamide gel electrophoresis. At 2 weeks of age collagen type III represented about one-third of types I and III in all tissues. After this age the proportion of type III increased up to 1 year in both heart (53.9 +/- 1.8%) and lung (47.7 +/- 2.8%), whereas it decreased in skin (18.6 +/- 1.5%). These data show that there are age-related changes in both collagen concentration and the relative proportions of types I and III collagen, but that the direction of these changes differs between tissues.

Thrombin stimulates smooth muscle cell procollagen synthesis and mRNA levels via a PAR-1 mediated mechanism.

Thrombin is a serine protease involved in haemostasis which exerts a number of cellular effects, including stimulating mesenchymal cell migration, proliferation, and has been implicated both in normal wound healing and pathological conditions associated with hyperproliferation of smooth muscle cells such as atherosclerosis and restenosis. We hypothesize that thrombin, in addition to its proliferative effects, may also influence the deposition of matrix proteins at sites of vascular injury by directly stimulating smooth muscle cell procollagen production. 10 nM thrombin significantly stimulated rat aortic smooth muscle cell procollagen production by 34 +/- 3% compared to media control cells over a 48 h incubation period, and increased steady state alpha1(I) procollagen mRNA levels by up to 104 +/- 22%. These effects are mediated via interaction of thrombin with the PAR-1 receptor since TRAP (Thrombin Receptor Activating Peptide) stimulated procollagen production by 23 +/- 0.5%. In addition, conditioned medium from thrombin-treated cells stimulated procollagen production by 30 +/- 3% suggesting that thrombin is acting via the production and/or release of an autocrine mediator. These data suggest a novel role for thrombin in vascular wound healing and the development of pathological conditions associated with increased connective tissue deposition.

Bleomycin-induced lung injury in the rat: effects of the platelet-activating factor (PAF) receptor antagonist BN 52021 and platelet depletion.

Bleomycin is a highly effective antitumor agent, but pulmonary toxicity, characterized by an acute inflammatory reaction and associated pulmonary edema, limits clinical use of the drug. Platelets and platelet-activating factor (PAF), a membrane-derived phospholipid, have been implicated in the mechanisms that can mediate pulmonary microvascular injury. We sought to investigate the role of PAF in bleomycin-induced lung injury in the rat, using the PAF receptor antagonist BN 52021; and the role of platelets though the use of an anti-platelet antibody. Lung injury was induced by intratracheal bleomycin (1.5 mg) and assessed by measurements of lung wet weight and total pulmonary extravascular albumin space (TPEAS). Bleomycin caused a significant increase in both indices after 48 hr, compared with control animals (p less than 0.05). A single dose of BN 52021 (20 mg/kg orally) significantly reduced the bleomycin-induced increase in lung weight, but not the rise in TPEAS (p greater than 0.05). Increasing the dose of BN 52021 (20 mg/kg/12 hr, orally) had no additional effect. Reducing circulating platelet numbers by approximately 75% had no effect on either the increase in lung weight or TPEAS, observed 48 hr after bleomycin (p greater than 0.05). PAF may partially contribute to the acute inflammatory reaction seen after intratracheal bleomycin in rats.

Transpulmonary gradient of type III procollagen peptides: acute effects of cardio-pulmonary bypass.

Type III procollagen N-peptides (PIIINPs) are believed to be released in stoichiometric amounts as type III collagen molecules are secreted from cells. We hypothesized that if the human lung actively produces type III collagen a detectable transpulmonary gradient in PIIINPs would exist in normal individuals that might be altered following a pulmonary insult. PIIINPs were therefore measured by radioimmunoassay in serum taken simultaneously from the pulmonary artery (PA) and left ventricle/aorta (LV) in 11 patients undergoing routine cardiac catheterisation. Mean PIIINP levels +/- SEM in LV were 66.8 +/- 5.4 micrograms.ml-1 and 59.9 +/- 4.1 micrograms.ml-1 in PA (p less than 0.04). In 6 patients, repeat measurements taken 4 h after cardiopulmonary bypass revealed a significant fall in PA values to 43.8 +/- 2.6 micrograms.ml-1 (p less than 0.001) and abolition of the transpulmonary gradient. These results suggest the adult human lung actively synthesis type III collagen and that, in the short term, cardiopulmonary bypass inhibits this process.

Long-term changes in mouse lung following inhalation of a fibrosis-inducing dose of 239PuO2: changes in collagen synthesis and degradation rates.

Mice were exposed by nose-only inhalation to 239PuO2, which resulted in an IAD of 1110 +/- 29 Bq. At various times after exposure, rates of collagen metabolism were measured using validated in vivo methods based on the administration of radiolabelled proline, together with a large flooding dose of unlabelled proline and measurement of its incorporation into lung collagen as hydroxyproline. Dramatic increases in both synthesis and degradation rates of collagen were observed. At 54 days after exposure the fractional synthesis rates in experimental mice were almost five times those in controls (control: 3.2 +/- 0.6%/day, 239PuO2-exposed: 14.5 +/- 0.4%/day) and by 300 days synthesis rates, although declining, were still more than double the control values. A similar pattern of change was observed for collagen degradation. The combination of changes in synthesis and degradation rates led to a 60% increase in lung collagen content by 300 days (control: 3.05 +/- 0.24 mg/lung, 239PuO2-exposed: 4.88 +/- 0.42 mg/lung). The data suggest that extensive remodelling of the lung connective tissue matrix occurs during development of fibrosis and that, over long periods of time, small imbalances between synthesis and degradation may result in quite large increases in protein content.

Effects of a parenteral supplement of folic acid and its interaction with level of feed intake on hepatic tissues and growth performance of young dairy heifers.

Forty-seven dairy heifers of approximately 10 d of age were assigned to a factorial experiment in which a supplement of folic acid (0 or 40 mg) administered weekly by i.m. injection and level of feed intake were the two factors studied. The heifers were weaned after 5 wk of experimentation. Following weaning, and until the end of the experiment, 11 wk later, they had ad libitum access to grass hay and concentrates at two different levels, ad libitum or restricted, to allow a body weight gain of 700 g/d. A supplement of folic acid (P less than .05) and ad libitum access to feed (P less than .05) increased the mean concentration of serum folates. Blood hemoglobin and packed cell volume were not affected by the level of feed intake. However, they were both increased (P less than .05) by the supplement of folic acid. Average daily gain was analyzed over three different periods: 0 to 5 wk (before weaning), 5 to 10 wk, and 10 to 16 wk. Average daily gain was increased by the supplement of folic acid during the second period (P less than .05) and by ad libitum access to feed during the last two periods (P less than .05). Ad libitum access to feed increased (P less than .05) weight of the liver, decreased the (P less than .05) concentrations of RNA and DNA, and increased (P less than .05) the ratios of protein/DNA and RNA/DNA. The supplement of folic acid decreased (P less than .05) weight of the liver and increased the ratio RNA/DNA (P less than .05). These effects of supplement of folic acid on growth performance and on hematological cells may reflect a lack of folic acid during the weeks after weaning.

Pulmonary gene therapy. Realistic hope for the future, or false dawn in the promised land?

Pulmonary gene therapy offers the hope of treatment for conditions such as cystic fibrosis, lung cancer, pulmonary fibrosis and acute respiratory distress syndrome for which current therapy is inadequate. Although initial clinical trials in cystic fibrosis and non-small cell lung cancer have shown promise the results have not been as good as might have been anticipated. However, clinical improvement has been demonstrated in conditions such as haemophilia [82], cardiovascular disease [83], head and neck cancer [84] and X-linked severe combined immunodeficiency disease [85]. The lack of success of pulmonary gene therapy is due, in part on the physical barriers to transfection perfected by the lung to prevent toxicity from inhaled particles, and partly due to the poor transfection efficiency of non-viral systems, and the immunogenicity of viral systems, of gene transfer. The LID vector goes some way to addressing the problems associated with current gene delivery strategies. With continued improvements in the properties of both viral and non-viral gene delivery systems leading to improved transfection efficiency with reduced toxicity, as well as the development of strategies aimed at reducing the physical barriers to pulmonary transfection, and targeting gene delivery systems to the site of injury, it is likely that pulmonary gene therapy will be used successfully to ameliorate a number of devastating pulmonary conditions.