SP-C Content Research Articles

Distillation of treated dairy effluent using integrated solar dish collector and solar still: A case study of distillate output enhancement

A solar thermal distillation system was designed to be compatible with electrically powered conventional laboratory distillation assembly. Its efficacy was checked in converting the treated dairy effluent into distilled water. The experimental setup comprised of a solar parabolic dish collector (aperture area 4.15 m2) and a solar still (basin area 1 m2) and was designed such that both can be used separately or in the combination. The study was conducted during March – June 2023. The solar dish collector was tested with two receivers with different bottom materials viz., stainless-steel and brass, thereby an average daily yield of 6.5 litres and 6.9 litres were obtained for both receivers, respectively. Simultaneously, solar still produced 2.08 litres of water individually per day. Thus, the highest yield obtained for whole setup was 8.9 litres per day considering individual performances of both the systems. The daily average thermal efficiency of dish collector for was found 65.59 % with steel receiver and 63.33 % with brass receiver. Similarly, thermal efficiency of solar still was 35 %. The quality of distillate was analysed on 10 parameters viz., pH, TDS, TSS, COD, BOD, Oil & Grease, SPC, Coliform count, electrical conductivity, and chloride content which showed promising results.

Modifying humus-phosphorus-arsenic interactions in sludge composting: The strengthening of phosphorus availability and arsenic efflux detoxification mechanisms

Arsenic (As) in sewage sludge poses a significant threat to environmental and human health, which has attracted widespread attention. This study investigated the value of adding sodium percarbonate (SP) on phosphorus (P) availability and As efflux detoxification through HS-P-As interactions. Due to the unique structure of humus (HS) and the similar chemical properties of P and As, the conditions for HS-P-As interaction are provided. This study discussed the content, morphology and microbial communities of HS, P and As by using metagenomic and correlation analysis. The results showed that the humification index in the experiment group (SPC) was 2.34 times higher than that in the control group (CK). The available phosphorus (AP) content of SPC increased from 71.09 mg/kg to 126.14 mg/kg, and SPC was 1.11 times that of CK. The relative abundance of ACR3/ArsB increased. Pst, Actinomyces and Bacillus commonly participated in P and As conversion. The correlation analysis revealed that the humification process was enhanced, the AP was strengthened, and the As was efflux detoxified after SP amendment. All in all, this study elucidated the key mechanism of HS-P-As interaction and put forward a new strategy for sewage sludge resource utilization and detoxification.

Influences of dietary soy protein concentrate with taurine on growth and biochemical status of Acanthopagrus schlegelii juveniles

Taurine is a component that tends to be deficient in fish diets that contain higher plant‐derivative proteins. Acanthopagrus schlegelii juveniles (IBW − 11.28 g) were fed nine diets containing SPC levels (SPC40, SPC60, SPC80 and SPC100), or supplemented with 5 g/kg of taurine at each replacement level. Our results showed higher WGR and SGR in SPC60 + TAU (WGR; 224.67, SGR; 2.10) followed by SPC40 + TAU (WGR; 221.75, SGR; 2.09). Phosphorus retention efficiency was significantly raised as the SPC increased when compared with the control diet. Taurine addition showed an incline in the nitrogen retention efficiency. An effect was observed within FR between fish and the SPC content. ADCs of dry matter, crude lipid and phosphorous were affected by SPC, while taurine addition affected only crude lipid. A significant effect was seen in ash and phosphorus by the SPC and taurine. TAU supplementation showed an increased T‐AOC activity in the SPC80 + TAU (1.90), while malondialdehyde and catalase activity were reduced except in completely replaced fish meal groups. It could be concluded that black sea bream can accept the substitution of FM with 60% of SPC while at the inclusion of dietary TAU (5 g/kg) growth could be improved.

In-situ solidification/stabilization of heavy metals contaminated site soil using a dry jet mixing method and new hydroxyapatite based binder.

In this study, in-situ treatment using dry jet mixing construction method and SPC (single superphosphate and calcium oxide) new binder are used to solidify/stabilize a heavy metal contaminated site soil with relatively high content of organic matters. Time-dependent field performance of the soils at 41 and 326 days after treatment is evaluated, which includes electrical conductivity (EC), leachability of heavy metals and chemical oxygen demand (COD), soil penetration resistance, acid neutralization capacity (ANC), and chemical speciation of heavy metals. The results indicate that the stabilized soils exhibit satisfactory performance which is comparable with the laboratory study. In-situ SPC treatment significantly decreases EC values and increases penetration resistance values of the soils. Leachability of lead, zinc, cadmium and COD decreases with increasing SPC content or curing time. Large percentages of heave metals in the soils are transformed from exchangeable fractions to residual fractions after treatment. These, coupled with the improved ANC, result in low heavy metal leachability in stabilized soils.

Hyperoxia induces the apoptosis of alveolar epithelial cells and changes of pulmonary surfactant proteins.

To explore the changes of surfactant protein C, D (SP-C, SP-D) and apoptosis of alveolar epithelial cells (AEC) in the lung injury of neonatal rats induced by hyperoxia. We divided neonatal rats within 24 hours into two groups randomly: the air group (n = 50) and the hyperoxia group (n = 50). Rats in the air group and hyperoxia group were bred conventionally and in tanks with normal pressure and 90% concentration of oxygen, respectively. On the 1st, 3rd, 7th, 10th, and 14th day after exposure, lung tissue of 8 rats in each group was collected and stained with hematoxylin and eosin (HE). We observed and recorded pathologic changes of lung tissue and detected apoptosis rate of alveolar epithelial cells by TUNEL (TdT-mediated dUTP nick end labeling). We detected the content of SP-C, SP-D in bronchoalveolar lavage fluid (BALF) by ELISA (enzyme-linked immunosorbent assay). In the air group, the alveolar formed gradually with equable size and regular shape during growth. While the amount of alveolar decreased gradually and we observed small vessels dilation and increasing hemorrhage as well as increasing interstitial cells and swollen lung tissue in the hyperoxia group. In the air group, the content of SP-C in the BALF reduced during growth. However, the content of SP-C in the hyperoxia group was lower than that in the air group on the first day, higher on the third day, reached the peak on the seventh day and began to decrease on the tenth day and more obviously on the fourteenth day. The level of SP-D in the air group declined gradually with growing. On the first day, the content of SP-D in the hyperoxia group was similar to that in the air group. It began to increase on the third day, reached its peak on the seventh day, and began to decrease on the tenth day, more obviously on the fourteenth day. Long-term exposure to hyperoxia inhibits the development of alveolar. The apoptosis of alveolar epithelial cells increased and the content of SP-C, SP-D in the lung tissue first increased and then decreased with the increase of exposure time.

Stabilization and solidification of a heavy metal contaminated site soil using a hydroxyapatite based binder

Synthetic hydroxyapatite (HA) is an efficient and environment-friendly material for the remediation of heavy metal contaminated soils. However, the application of conventional HA powder in stabilizing contaminated soils is limited, due to high cost of final products, difficulties in synthesizing purified HA crystals. A new binder named SPC, which composes of single superphosphate (SSP) and calcium oxide (CaO), is presented as an alternative in this study. HA can form in the soil matrix by an acid-base reaction between SSP and CaO, resulting in a dense structure and improved mechanical properties of treated soils. Therefore, the SPC is capable of effectively immobilizing heavy metals and elevating strength of contaminated soils, meanwhile, maintaining relatively low cost. This paper presents a systematic investigation of the performance, reaction products, and microstructural properties of a lead (Pb), zinc (Zn), and cadmium (Cd) contaminated industrial site soil stabilized with SPC binder. The effects of SPC content and curing time on the pH, leachability and strength properties of the stabilized soils are evaluated. Furthermore, modified European Communities Bureau of Reference (BCR) sequential extraction procedure (SEP), mercury intrusion porosimetry (MIP), X-ray diffraction (XRD), and scanning electron microscope (SEM) analyses are performed to interpret the mechanisms controlling the changes in these macro-properties. The results show that the soil pH and unconfined compressive strength (UCS) increase with increasing SPC content and curing time. After 28days of curing, the UCS values of stabilized soils are approximately 2.2–5.7 times those of the untreated soil. The leachability of Pb, Zn and Cd is significantly reduced after stabilization, and the SPC content and curing time have considerable influences on the leached concentrations of heavy metals. The SEP results confirm that SPC significantly reduces the acid soluble fractions of Pb, Zn and Cd while increases their residual fractions. The MIP test results show that pore volume reduces notably and pore profile of the soil changes remarkably after SPC stabilization. The mineralogical (XRD) and microstructural (SEM) analyses reveal that the formation of heavy metal-bearing hydroxyapatites and phosphate-based precipitates are the primary mechanisms of immobilization of Pb, Zn and Cd in the SPC stabilized soil.

The change of pulmonary surfactant protein of rat following silica exposure

To investigate the change of lung surfactant protein (SP) A,B,C,D of rats following silica dust exposure in order to provide the evidences for the early diagnosis indices or therapy of silicosis. 60 male SD rats were randomly divided into silica group, and corresponding controls group. Rats in silica group were administrated 1 ml silica solution by intratracheal instillation at dose of 50 mg/ml. Rats in control group were administrated the same amount saline. At 3rd, 7th, 14th, 21st, 28th after silica exposure, serum and bronchoalveolar lavage fluid (BALF) samples were obtained. The concentration of SP-A, SP-B, SP-C, SP-D in serum and BALF were measured by using enzyme immunoassay (ELISA). Meanwhile the levels of total anti-oxidative activity (T-AOC) and hydroxyproline (HYP) in lung tissue were also detected. The pathology of lung tissue was conducted. Compared with control group, SP-A concentration in BALF of silica exposed rat for 3, 14, 21, 28d was significant lower and SP-D concentration in BALF of silica exposed rat for all time points was also lower. The differences were significant (P < 0.05). Meanwhile SP-B level in 7, 14, 21, 28 d silica exposed rats BALF and SP-C level in 14, 21, 28 d silica exposed rats markedly decreased (P < 0.05). In addition compared with control group, SP-A, SP-B and SP-C concentration in serum of silica exposed rat were higher when SP-A for 14, 21, 28 d silica exposure, SP-B for 7, 14, 21 d silica exposure and Sp-C for 7, 14, 21, 28 d exposure. And all difference were significant (P < 0.05). As silica exposure time increased, SP-C concentration in serum showed an increase trend, which showed a time-response relationship (r = 0.618, P = 0.042). However, SP-D concentration in serum of rat for 7, 14, 21, 28d silica exposure were significant lower than that of control group (P < 0.005). And there was a decrease trend with time point exposure regarding of SP-D (r = -0.731, P = 0.016). The HYP content in lung tissue of experiment rats increased at 3rd, 7th, 14th, 21st and 28th day time point and The T-AOC activity in lung tissue decrease at, 7th, 14th, 21st and 28th day time point. The differences were significant (P < 0.05). There was a positive correlation (P = 0.803, P = 0.045) between SP-C in BALF and HYP of silica exposed rats and a negative correlation between SP-D in BALF and HYP (r = -0.867, P = 0.033). No significant correlation were seen between SP-A, SP-B BALF and HYP (y = 0.416, P = 0.28; r = 0.592, P = 0.071). SP-C concentration in BALF and serum all showed an increased trend and a positive correlation was seen (r = 0.539, P = 0.046). The same decrease trend was seen between SP-D in BALF and serum and correlation value was 0.870 (P = 0.034). The silica exposure did cause the change of SP content both in BALF and serum. The SP-C and SP-D content in serum might be served as an early effective biomarker of silicosis.

French summaries of product characteristics: content in relation to therapeutic monitoring of psychotropic drugs

The prescription information (summary of product characteristics, SPC) is compiled by the pharmaceutical industry as required by the national regulatory authorities. They vary in their content about the properties of drugs and about the usefulness of therapeutic drug monitoring (TDM) in the blood of patients. Based on a previous study carried out in Germany, the degree of agreement of French SPC for 59 psychotropic drugs with the existing medico-scientific evidence in the area of TDM was examined using a recently developed instrument. A summary score of SPC content (SPCC) related to TDM (SPCC(TDM)) has been calculated and compared with the level of recommendation of TDM of the AGNP-TDM expert group consensus guidelines for TDM in psychiatry [AGNP: Arbeitsgemeinschaft für Neuropsychopharmakologie und Pharmakopsychiatrie (Association for neuropsychopharmacology and pharmacopsychiatry)]. Among the antidepressants, antipsychotics, tranquillizers/hypnotic agents and mood stabilizers, the highest SPCC(TDM) scores in the French SPC were reached for imipramine (16), haloperidol (6), clonazepam (8) and lithium (23), respectively. Results were similar to those obtained from the analysis of German SPC, and considerable disagreement was found between the information on TDM in SPC and existing medico-scientific evidence, albeit less in the case of mood stabilizers. Taking into account the recommendations of the AGNP-TDM expert group guidelines, there is a deficit in the French SPC concerning TDM-relevant information. An amelioration of this situation could help to improve the clinical practice of TDM of psychotropic drugs, as the SPC is a widely used tool.

Inactivation of pulmonary surfactant by silicone oil in vitro and in ventilated immature rabbits

Surface activity of pulmonary surfactant is impaired by exposure to syringes lubricated with silicone oil (SO). These syringes are used daily in clinical practice. In vitro experiments were used for detection of SO, determination of surface activity, and semiquantitative measurement of surfactant protein (SP)-B and -C in SO/surfactant mixtures. Randomized, controlled animal studies were applied for determination of in vivo activity. University research laboratory. Mass spectrometry of SO originating from syringes with and without surfactant was performed. The surface activity of SO plus surfactant phospholipids (PLs) or modified natural surfactant (Curosurf) was measured. SO/Curosurf preparations were further analyzed for changes in the content of SP-B and SP-C using immunoblotting. Neonatal rabbits received mixtures of SO/Curosurf (ratio 0-1.3 mg/mg PL) intratracheally and were then ventilated with a standardized sequence of peak insufflation pressures. Tidal volume curves were recorded, gas volumes of excised lungs were measured, and histologic analysis was performed. Dissolved SO was found after rinsing syringes with organic solvents or Curosurf. Surface activity of Curosurf was significantly reduced after addition of 0.13-1.3 mg SO/mg PL. Immunoblotting revealed interference of SO with SP-B, but not with SP-C. With increasing SO/Curosurf ratios, patchy alveolar air expansion was observed, lung gas volumes were reduced, and time to inflate the lungs was increased, whereas compliance and tidal volumes remained unimpaired. In vitro SO impairs surface activity of Curosurf and leads to interference with SP-B. SO contamination of exogenous surfactant impairs lung function in animal studies and should be avoided.

Value and Actuality of the Prescription Information for Therapeutic Drug Monitoring of Psychopharmaceuticals: A Comparison with the Medico-Scientific Evidence

Therapeutic drug monitoring (TDM) of psychopharmaceuticals, i.e., the assay of plasma concentrations, is a practical therapeutic application of pharmacokinetic principles in psychiatry. The prescription information (summary of product characteristics, SPC) is provided by pharmaceutical companies according to the requirements of regulatory authorities. The present study investigated the degree of agreement of German SPCs for 48 psychopharmaceuticals with the existing medico-scientific evidence in the area of TDM. For this aim, an empirical summary score of SPC content related to TDM (SPCC (TDM)) was calculated and compared with the level of recommendation of TDM (LOR) of the AGNP-TDM expert group consensus guidelines. Considerable disagreement was found between the information on TDM in SPCs and existing medico-scientific evidence, e.g., in the case of antidepressant and antipsychotic drugs. Even for well studied compounds, such as amitriptyline and clozapine, insufficient information on TDM is included in German SPCs. Small differences existed in the TDM-related information in SPCs of generic drugs with, however, much variance between Germany, Austria and Switzerland. Generally, it must be concluded that deficits exist in the preparation of German SPCs for psychopharmaceutical drugs with respect to empirical pharmacokinetic data, i.e., TDM-relevant information. It is recommended that SPCs of psychopharmaceuticals should be improved in terms of TDM-related information and that target plasma concentrations be adjusted according to the guidelines of the AGNP-TDM expert group. A higher level of good pharmacokinetic practice may be thus achieved.

Pulmonary surfactant in patients with Pneumocystis pneumonia and acquired immunodeficiency syndrome

Pneumocystis pneumonia (PCP) is a severe infection of the immunocompromised host, resulting in diffuse alveolar damage and life-threatening respiratory failure. We analyzed pulmonary surfactant composition and function in bronchoalveolar lavage fluid (BALF) from ventilated and spontaneously breathing HIV-positive patients with PCP. Prospective clinical trial. University hospital intensive care unit. Thirty-four spontaneously breathing (SB-PCP) and 20 ventilated HIV-positive patients with PCP (V-PCP), ten patients with acute respiratory distress syndrome (ARDS), 11 spontaneously breathing patients with bacterial pneumonia (PNEU), and 22 healthy volunteers. None. Total phospholipid in BALF did not differ between any category vs. controls, whereas total protein increased approximately 14-fold in V-PCP and five-fold in SB-PCP compared with controls (p < .001). The relative content of large surfactant aggregates (LA) was reduced in SB-PCP and V-PCP compared with controls (p < .05). The phospholipid and fatty acid profiles showed a significant reduction in the relative content of phosphatidylcholine (PC), phosphatidylglycerol, and palmitic acid in PC in all patient categories compared with controls, with more in V-PCP (p < .001) compared with SB-PCP (p < .05). The neutral lipid-to-phospholipid ratio in LA was three-fold elevated in V-PCP (p < .01 compared with control) but not in SB-PCP. Analysis of neutral lipid classes showed a significant increase in the relative content of triglycerides and a reduction in free fatty acids in V-PCP compared with controls. BALF surfactant protein (SP)-A and SP-D significantly increased in V-PCP and SB-PCP, but not in ARDS and PNEU, compared with controls (p < .05). SP-B and SP-C content in LA remained unchanged in PCP compared with controls but decreased significantly in ARDS and PNEU. The minimum surface tension of LA was impaired (p < .001) in V-PCP more than in SB-PCP and was strongly correlated with the reduction in palmitic acid levels in PC LA (r = -.81). Reductions in phosphatidylglycerol strongly correlated with decreased Pao2/Fio2 values (r = .72). We conclude that severe alterations in surfactant function and composition occur in patients with PCP and are even more pronounced in ventilated patients than in nonventilated patients. Surfactant lipid changes in PCP, but not surfactant protein profiles, closely resemble those found in ARDS.

Lipidomics of cellular and secreted phospholipids from differentiated human fetal type II alveolar epithelial cells

Maturation of fetal alveolar type II epithelial cells in utero is characterized by specific changes to lung surfactant phospholipids. Here, we quantified the effects of hormonal differentiation in vitro on the molecular specificity of cellular and secreted phospholipids from human fetal type II epithelial cells using electrospray ionization mass spectrometry. Differentiation, assessed by morphology and changes in gene expression, was accompanied by restricted and specific modifications to cell phospholipids, principally enrichments of shorter chain species of phosphatidylcholine (PC) and phosphatidylinositol, that were not observed in fetal lung fibroblasts. Treatment of differentiated epithelial cells with secretagogues stimulated the secretion of functional surfactant-containing surfactant proteins B and C (SP-B and SP-C). Secreted material was further enriched in this same set of phospholipid species but was characterized by increased contents of short-chain monounsaturated and disaturated species other than dipalmitoyl PC (PC16:0/16:0), principally palmitoylmyristoyl PC (PC16:0/14:0) and palmitoylpalmitoleoyl PC (PC16:0/16:1). Mixtures of these PC molecular species, phosphatidylglycerol, and SP-B and SP-C were functionally active and rapidly generated low surface tension on compression in a pulsating bubble surfactometer. These results suggest that hormonally differentiated human fetal type II cells do not select the molecular composition of surfactant phospholipid on the basis of saturation but, more likely, on the basis of acyl chain length.

Reversibility of lung inflammation caused by SP-B deficiency

Whereas decreased concentrations of surfactant protein (SP)-B are associated with lung injury and respiratory distress, potential causal relationships between SP-B deficiency and lung inflammation remain unclear. A transgenic mouse in which human SP-B expression was placed under conditional control of doxycycline via the CCSP promoter was utilized to determine the role of SP-B in the initiation of pulmonary inflammation. Adult mice, made SP-B deficient by removal of doxycycline, developed severe respiratory failure within 4 days. Deficiency of SP-B was associated with increased minimal surface tension of the surfactant and perturbed lung mechanics. Four days of SP-B deficiency did not alter SP-C content or surfactant phospholipid content or composition. SP-B deficiency was associated with lung inflammation and increased soluble L-selectin, STAT-3, and phosphorylated STAT-3 in alveolar macrophages and alveolar epithelial cells. Alveolar IL-6, IL-1beta, and macrophage inflammatory protein-2 concentrations were increased after removal of doxycycline, indicating pulmonary inflammation. Restoration of SP-B expression following administration of doxycycline rapidly reversed SP-B-dependent abnormalities in lung mechanics and inflammation. SP-B deficiency is sufficient to cause lung dysfunction and inflammation in adult mice. SP-B reversed inflammation and maintained lung function in vivo, indicating its potential utility for the prevention and treatment of pulmonary injury and surfactant deficiency.

Dysfunction of Pulmonary Surfactant in Chronically Ventilated Premature Infants

Infants of <30 wk gestation often require respiratory support for several weeks and may develop bronchopulmonary dysplasia (BPD), which is associated with long-term pulmonary disability or death in severe cases. To examine the status of surfactant in infants at high risk for BPD, this prospective study analyzed 247 tracheal aspirate samples from 68 infants of 23-30 wk gestation who remained intubated for 7-84 d. Seventy-five percent of the infants had one or more surfactant samples with abnormal function (minimum surface tension 5.1-21.7 mN/m by pulsating bubble surfactometer), which were temporally associated with episodes of infection (p = 0.01) and respiratory deterioration (p = 0.005). Comparing normal and abnormal surfactant samples, there were no differences in amount of surfactant phospholipid, normalized to total protein that was recovered from tracheal aspirate, or in relative content of phosphatidylcholine and phosphatidylglycerol. Contents of surfactant proteins (SP) A, B, and C, measured in the surfactant pellet by immunoassay, were reduced by 50%, 80%, and 72%, respectively, in samples with abnormal surface tension (p < or = 0.001). On multivariable analysis of all samples, SP-B content (r = -0.58, p < 0.0001) and SP-C content (r = -0.32, p < 0.001) were correlated with surfactant function. We conclude that most premature infants requiring continued respiratory support after 7 d of age experience transient episodes of dysfunctional surfactant that are associated with a deficiency of SP-B and SP-C.

Kinetics of phospholipid insertion into monolayers containing the lung surfactant proteins SP-B or SP-C.

The lung surfactant proteins SP-B and SP-C are pivotal for fast and reversible lipid insertion at the air/liquid interface, a prerequisite for functional lung activity. We used a model system consisting of a preformed monolayer at the air/liquid interface supplemented with surfactant protein SP-B or SP-C and unilamellar vesicles injected into the subphase of a film balance. The content of SP-B or SP-C was similar to that found in lung lavage. In order to elucidate distinct steps of lipid insertion, we measured the time-dependent pressure increase as a function of the initial surface pressure, the temperature and the phosphatidylglycerol content by means of surface tension measurements and scanning force microscopy (SFM). The results of the film balance study are indicative of a two-step mechanism in which initial adsorption of vesicles to the protein-containing monolayer is followed by rupture and integration of lipid material. Furthermore, we found that vesicle adsorption on a preformed monolayer supplemented with SP-B or SP-C is strongly enhanced by negatively charged lipids as provided by DPPG and the presence of Ca2+ ions in the subphase. Hence, long-range electrostatic interactions are thought to play an important role in attracting vesicles to the surface, being the initial step in replenishment of lipid material. While insertion into the monolayer is independent of the type of protein SP-B or SP-C, initial adsorption is faster in the presence of SP-B than SP-C. We propose that the preferential interaction between SP-B and negatively charged DPPG leads to accumulation of negative charges in particular regions, causing strong adhesion between DPPG-containing vesicles and the monolayer mediated by Ca2+ ions, which eventually causes flattening and rupture of attached liposomes as observed by in situ SFM.

Mechanical distension modulates pulmonary alveolar epithelial phenotypic expression in vitro.

The pulmonary alveolar epithelium is composed of two distinct types of cells, type I and type II cells, both of which are critical for normal lung function. On the basis of experiments of both nature and in vivo studies, it has been hypothesized that expression of the type I or type II phenotype is influenced by mechanical factors. We have investigated the effects of mechanical distension on the expression of specific markers for the type I and type II cell phenotypes in cultured alveolar type II cells. Rat alveolar type II cells were tonically mechanically distended in culture. Cells were analyzed for a marker for the type I phenotype (rTI40, an integral membrane protein specific for type I cells) and for markers for the type II phenotype [surfactant protein (SP) A, SP-B, and SP-C] as well as for glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Mechanical distension caused a 68 +/- 25% (n = 3) increase in mRNA content of rTI40 relative to undistended controls. In contrast, mechanical distension resulted in a decrease in mRNA content of SP-B to 35 +/- 19% (n = 3) and of SP-C to 20 +/- 6.7% (n = 3) of undistended controls. There was no effect on mRNA content of SP-A or GAPDH. The differences in mRNA content of SP-B and SP-C were found to be primarily due to changes at the transcriptional level by nuclear run-on assays. The effects on rTI40 appear to be due to posttranscriptional events. These data show that mechanical distension influences alveolar epithelial phenotypic expression in vitro, at least in part, at the transcriptional level.

ADDITION OF SYNTHETIC SURFACTANT PROTEIN B TO SURVANTA IMPROVES GAS EXCHANGE IN SURFACTANT-DEFICIENT RATS. † 2114

Surfactant protein B (SP-B) is critical to surfactant function. Survanta®, a modified bovine lung extract, contains less SP-B than surfactants derived from lung lavage (Seeger et al., Eur Resp J 1993;6:971). Supplementation of Survanta with native SP-B increases dynamic compliance and deflation lung volume in preterm rabbits (Mizuno et al., Pediatr Res 1995;37:271). We hypothesized that the in vivo function of Survanta can be improved by supplementation with synthetic SP-B and SP-C and tested this hypothesis in a surfactant-deficient rat model. Full length SP-B1-78 and palmitoylated SP-C1-34 were synthesized, purified, derivatized, and analyzed by mass spectrometry and amino acid analysis. Survanta was extracted, unmodified (Surv-1) or enriched with 1% SP-B (Surv-2), 2% SP-B (Surv-3), or 2% SP-B + 1% SP-C (Surv-4), and resuspended. In vitro surface activity was tested on a modified Langmuir/Wilhelmy balance and SP-B and SP-C content were measured by Fourier transform infrared spectroscopy. Adult rats were anesthetized, tracheotomized, had an arterial line placed, and were ventilated with 100% oxygen, a peak inspiratory pressure of 30 cm H2O and a rate of 40/min. The lungs were lavaged with warmed normal saline until the PaO2 dropped below 60 torr, at which time 100 mg/kg of one of the four surfactant preparations was instilled. After 15 min of ventilation, the rats were killed and pressure-volume curves performedin situ. The mean arterial/Alveolar PO2 ratio was 0.08 post-lavage and increased by 74% after treatment with Surv-1 (n=5), by 92% after treatment with Surv-2 (n=6), by 257% after treatment with Surv-3 (n=5), and by 202% after treatment with Surv-4 (n=6). Surv-4 not only improved PaO2, but also reduced PaCO2 values. These data suggest that the addition of 2% synthetic SP-B to Survanta improves its short-term positive effects on oxygenation in this surfactant-deficient rat model.

Solubility of hydrophobic surfactant proteins in organic solvent/water mixtures. Structural studies on SP-B and SP-C in aqueous organic solvents and lipids

The solubility of hydrophobic pulmonary surfactant proteins in different organic solvents and organic solvent/water combinations has been analyzed. Three organic solvents have been selected: methanol (MetOH), acetonitrile (ACN) and trifluorethanol (TFE). Porcine SP-B showed very similar calculated secondary structure when dissolved in methanol, 60% ACN or 70% TFE and reconstituted in lysophosphatidylcholine (LPC) micelles or dipalmitoylphosphatidylcholine (DPPC) vesicles, as deduced from circular dichroism studies. SP-B was calculated to posses around 45% of α-helix in all these systems. The fluorescence emission spectrum of SP-B has been also characterized in aqueous solvents and lipids. It always showed a splitting of the tryptophan contribution into two components with different emission maxima. SP-C had a very different structure in 80% ACN or 70% TFE. While α-helix was the main secondary structure of SP-C in ACN/water mixtures — around 50% -, it had almost exclusively β-structure when dissolved in 70% TFE. The CD spectrum of SP-C in TFE showed dependence on the protein concentration, suggesting that protein-protein interactions could be important in this β-conformation. SP-C reconstituted in LPC micelles or DPPC vesicles had a CD spectrum qualitatively similar to that one in aqueous ACN, with a dominant α-helical structure. The α-helical content of SP-C in micelles of LPC and vesicles of DPPC, 60 and 70%, respectively, was calculated to be higher than the α-helical content of the protein dissolved in any aqueous organic solvent.

Increased Expression of Pulmonary Surfactant Proteins in Oxygen-exposed Rats

Exposure of adult rats to 85% ambient oxygen increased the content of surfactant proteins SP-A, SP-B, and SP-C recovered from alveolar lavage. The surfactant proteins increased during 1 to 7 d of oxygen exposure. The increased surfactant protein was associated with increased relative abundance of mRNA encoding each of the proteins in lung tissue. Exposure to hyperoxia progressively increased the amounts of the surfactant proteins in alveolar lavage fluid as estimated by immunoblot analysis after sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The mRNAs encoding SP-A (1.7 and 1.0 kb), SP-B (1.6 kb), and SP-C (0.9 kb) increased significantly after oxygen exposure for 5 d. The present findings support the concept that oxygen exposure mediates surfactant protein expression at a pretranslational level.