ABSTRACT
Drip irrigation is a means of distributing the exact amount of water a plant needs by dripping water directly onto the root zone. It can produce up to 90% more crops than rain-fed irrigation, and reduce water consumption by 70% compared to conventional flood irrigation. Drip irrigation may enable millions of poor farmers to rise out of poverty by growing more and higher value crops, while not contributing to overconsumption of water. Achieving this impact will require broadening the engineering knowledge required to design new, low-cost, low-power drip irrigation technology, particularly for poor, off-grid communities in developing countries. For more than 50 years, pressure compensating (PC) drip emitters-which can maintain a constant flow rate under variations in pressure, to ensure uniform water distribution on a field-have been designed and optimized empirically. This study presents a parametric model that describes the fluid and solid mechanics that govern the behavior of a common PC emitter architecture, which uses a flexible diaphragm to limit flow. The model was validated by testing nine prototypes with geometric variations, all of which matched predicted performance to within R2 = 0.85. This parametric model will enable irrigation engineers to design new drip emitters with attributes that improve performance and lower cost, which will promote the use of drip irrigation throughout the world.
Subject(s)
Agricultural Irrigation , Crops, Agricultural , Models, TheoreticalABSTRACT
Female birds deposit or 'excrete' lipophilic contaminants to their eggs during egg formation. Concentrations of xenobiotics in bird eggs can therefore accurately indicate levels of contamination in the environment and sampling of bird eggs is commonly used as a bio-monitoring tool. It is widely assumed that maternally transferred contaminants cause adverse effects on embryos but there has been relatively little experimental work confirming direct developmental effects (cf. behaviorally-mediated effects). We validated the use of egg injection for studies of in ovo exposure to xenobiotics for a small songbird model species, the zebra finch (Taeniopygia guttata), where egg weight averages only 1 g. We investigated a) the effect of puncturing eggs with or without vehicle (DMSO) injection on egg fate (embryo development), chick hatching success and subsequent growth to 90 days (sexual maturity), and b) effects of two vehicle solutions (DMSO and safflower oil) on embryo and chick growth. PBDE-99 and -47 were measured in in ovo PBDE-treated eggs, chicks and adults to investigate relationships between putative injection amounts and the time course of metabolism (debromination) of PBDE-99 during early development. We successfully injected a small volume (5 µL) of vehicle into eggs, at incubation day 0, with no effects on egg or embryo fate and with hatchability similar to that for non-manipulated eggs in our captive-breeding colony (43% vs. 48%). We did find some evidence for an inhibitory effect of DMSO vehicle on post-hatching chick growth, in male chicks only. This method can be used to treat eggs in a dose-dependent, and ecologically-relevant, manner with PBDE-99, based on chemical analysis of eggs, hatchling and adults.
Subject(s)
Embryonic Development/drug effects , Environmental Monitoring/methods , Models, Biological , Ovum/drug effects , Teratogens/toxicity , Toxicity Tests/methods , Animals , Embryo, Nonmammalian/drug effects , Female , Finches , MaleABSTRACT
A volume replacement therapy compensates a reduced intravascular volume to stabilize and maintain hemodynamics and vital signs. For this therapy, a physiologically-based solution comprising both, osmotic and colloid osmotic components, should be administered. The basic requirement for a sufficient fluid replacement and volume resuscitation therapy in children are the profound and special knowledge of the physiological and pathophysiological interactions in water balance and electrolyte metabolism in childhood, the pharmacology of the applied solutions and the adequate monitoring of this fluid and volume replacement therapy. Wrong dosages and side effects are reasons for a negative postoperative outcome in children.
Subject(s)
Fluid Therapy/methods , Infusions, Parenteral/methods , Perioperative Care/methods , Age Factors , Child , Humans , Hypovolemia/prevention & control , Monitoring, PhysiologicABSTRACT
OBJECTIVE: Umbilical artery catheter (UAC) use is common in the management of critically ill neonates; however, little information exists regarding the anatomic and vascular effects of UAC placement in premature newborns. STUDY DESIGN: Baboons were delivered at 125 days of gestation (term=185 days), treated with surfactant, had UACs placed and were ventilated for either 6 or 14 days. Animals were assigned to short-term (6 days, n=6) and long-term (14 days, n=30) UAC placement. At necropsy, aortas were removed with UACs still in place. Histological examination of upper, middle and lower aorta specimens stained with hematoxylin and eosin and immunolabeled to detect smooth muscle (alpha-actin) was carried out in a blinded manner. Controls were delivered at 125, 140 and 185 days and the aortas acquired immediately after birth. None of the non-catheterized control animals (125 days, n=4; 140 days, n=5; and 185 days, n=5) had aortic vessel thrombi or vascular wall abnormalities. RESULT: All 6 animals with short-term (6/6, 100%) and 18 animals with long-term (18/30, 60%) UAC placement displayed aortic thrombi and neointimal proliferation of the vascular wall. The majority (60%) of analyzed animals with UAC placement displaying neointimal hyperplasia were immunopositive for alpha-actin, indicating the presence of smooth muscle in these lesions. CONCLUSION: Our findings suggest that both short- and long-term UAC use is associated with aortic wall pathological abnormalities compared with control animals. This study emphasizes the judicious use and early removal of UACs if possible in order to potentially prevent significant hemostatic and aortic wall vascular complications.
Subject(s)
Aortic Diseases/etiology , Catheterization/adverse effects , Disease Models, Animal , Infant, Premature, Diseases/etiology , Thrombosis/etiology , Umbilical Arteries , Animals , Animals, Newborn , Aortic Diseases/pathology , Female , Humans , Infant, Newborn , Infant, Premature , Infant, Premature, Diseases/pathology , Male , Papio , Thrombosis/pathologyABSTRACT
A structured and well-organized postgraduate professional education in medical doctors is an important and necessary component in health care these days. It is the foundation for a high quality and safety of physicians' daily work. The article describes the anesthesiological educational program, which is required to finish anesthesia residency in Germany successfully.
Subject(s)
Anesthesiology/education , Education, Medical, Continuing/methods , Teaching/methods , Anesthesiology/classification , Curriculum/standards , Germany , Program Evaluation , Terminology as TopicABSTRACT
Treatment of acute pain is one of the most important tasks of perioperative pediatric anesthesia. Pain-relieving drugs are usually administered on the basis of the concept of balanced analgesia, which involves a combination of analgesics with either synergic or additive effects. The concept of balanced analgesia is successful under many circumstances, but continuous efforts are needed to improve management in dificult cases. The key problem is to decide if a child has pain. Establishing a high standard at an institution requires a regular evaluation and documentation of pain scores. Non-opioid analgesics are frequently administered just in the induction of anesthesia due to their long-term effects, and then combined with opioids or regional anesthetic techniques. The present review focuses on non-opioid analgesics that can be intravenously injected for the perioperative treatment of acute pain.
Subject(s)
Anti-Inflammatory Agents, Non-Steroidal/therapeutic use , Narcotics/therapeutic use , Pain Measurement , Pain, Postoperative/drug therapy , Anesthesia, Caudal , Anesthesia, Conduction , Anti-Inflammatory Agents, Non-Steroidal/administration & dosage , Anti-Inflammatory Agents, Non-Steroidal/adverse effects , Child , Humans , Narcotics/administration & dosage , Narcotics/adverse effects , Nerve BlockSubject(s)
Blood Transfusion/statistics & numerical data , Hemodilution/methods , Preoperative Care/methods , Prostatic Neoplasms/surgery , Urinary Bladder Neoplasms/surgery , Blood Coagulation/drug effects , Blood Loss, Surgical/prevention & control , Blood Transfusion/economics , Central Venous Pressure/drug effects , Cystectomy , Hemodilution/adverse effects , Hemodilution/economics , Humans , Hydroxyethyl Starch Derivatives/adverse effects , Hydroxyethyl Starch Derivatives/economics , Hydroxyethyl Starch Derivatives/therapeutic use , Male , Plasma Substitutes/adverse effects , Plasma Substitutes/economics , Plasma Substitutes/therapeutic use , Preoperative Care/adverse effects , Preoperative Care/economics , Prospective Studies , Prostate/surgery , Prostatectomy , Treatment OutcomeABSTRACT
Severe sepsis with multiple organ dysfunctions is still the leading cause of death in non-cardiac intensive care units (ICU). The incidence is expected to rise in the future due to the growing number of older and immunocompromized patients, the use of invasive procedures, and an increase in the percentage of aggressive or resistant microorganisms. Despite the enormous investment in critical care resources, severe sepsis mortality remains to be high and ranges from 28 to 50%. Based on the disappointing experiences with anti-inflammatory strategies, the authors now realize that sepsis is more than just an inflammation. By taking into account pathophysiological changes, the new therapeutic concepts combine successful modulation with an improvement of the ICU management of multiple organ dysfunctions. This review discusses the actual sepsis intensive care concepts that may be useful in reducing unacceptable high mortality rates in patients with severe sepsis.
Subject(s)
Multiple Organ Failure/etiology , Multiple Organ Failure/therapy , Sepsis/etiology , Sepsis/therapy , Humans , Multiple Organ Failure/drug therapy , Multiple Organ Failure/surgery , Sepsis/drug therapy , Sepsis/surgeryABSTRACT
UNLABELLED: The efficacy of hypervolemic hemodilution (HHD) in reducing the rate of donor blood transfusion is controversely discussed. The present prospective, randomized, clinical study analyzes the impact of HHD with 6% hydroxyethyl starch (HES) 130/0.4 solution on the rate transfusion, laboratory parameters, and the incidence of complications as compared with those in the control group receiving no preoperative HHD. MATERIAL AND METHODS: 80 patients who had undergone total prostatectomy or cystectomy were randomized into 2 groups. Before anesthesia, the HHD group (n=40) received 15 ml/kg of 6% HES 130/0.4 solution. In the HHD and control (n=40) groups, 6% HES 130/0.4 was intraintraoperatively infused in its maximum dose of 33 ml/kg according to the patients' needs. Indicationsf or blood transfusion trigger were Hb <5 g/dl or packed cell volume < 0.24. Laboratory parameters (Hb, PCV, platelets, prothrombin index, prothrombin time, thrombin time, fibrinogen, antithrombin III were measured before surgery, after HHD and 2, 24, and 48 hours after surgery. The mean blood pressure (MBP), heart rate (HR), and central venous pressure (CVP) were determined before surgery, after HHD, and 2 hours postsurgery. Statistical measurements were made in all patients from the HHD and control groups, as well as in a subgroup, in which intraoperative blood loss was greater than 30% of the total blood volume (70 ml/kg body weight). RESULTS: Demographic data and surgical techniques were similar in both groups. After HHD, CVP rose significantly. The changes in MBP and HR were statistically insignificant. There were no complications after HHD. Of the 40 HHD-group patents, 5 needed donor blood transfusion while in the control group blood was transfused to 10 of the 40 patients in the control group. Comparing both groups showed a lower need for blood transfusion in the HHD-group patients than in the controls (10 versus 24 packed red cells). The postoperative Hb values showed no difference between both groups. CONCLUSIONS: Preoperative HHD in patients undergoing surgery with expected >30% blood loss leads to decreased needs for blood transfusion. The method is safe and easy-to-use.
Subject(s)
Blood Loss, Surgical/prevention & control , Blood Transfusion , Hemodilution/methods , Hydroxyethyl Starch Derivatives/administration & dosage , Plasma Substitutes/administration & dosage , Preoperative Care/methods , Humans , Male , Middle AgedABSTRACT
This study evaluated the relationship between neuropsychological and affective functioning, subjective sleepiness and sleep-disordered breathing in 100 patients with obstructive sleep apnea (OSA). Using principal components analysis, three indices of sleep-disordered breathing were identified from polysomnography: sleep disturbance, extent of nocturnal hypoxemia, and sleep quality. Poorer sleep quality was related to slower processing speed, somatic symptomatology and tension-anxiety levels. Nocturnal hypoxemia was related to visuconstructional abilities, processing speed and mental flexibility. Patients who had high levels of subjective sleepiness had poorer performances on a complex task of executive functioning and higher levels of tension-anxiety. These results imply a differential effect of sleep-disordered breathing on domains of neuropsychological functioning. Additionally, they suggest that a patient's subjective level of sleepiness is a good predictor of certain aspects of neurobehavioral functioning.
Subject(s)
Affect/physiology , Hypoxia/etiology , Memory, Short-Term/physiology , Reaction Time/physiology , Sleep Apnea, Obstructive/physiopathology , Sleep/physiology , Adult , Case-Control Studies , Female , Humans , Male , Middle Aged , Neuropsychological Tests/statistics & numerical data , Polysomnography/methods , Principal Component Analysis/methods , Problem Solving , Psychomotor Performance , Sleep Apnea, Obstructive/complications , Sleep Stages , Verbal Learning/physiology , Visual PerceptionABSTRACT
As Plasmodium rely extensively on homolactic fermentation for energy production, Plasmodium falciparum lactate dehydrogenase (PfLDH)--the key enzyme in this process--has previously been suggested as a novel target for antimalarials. This enzyme has distinctive kinetic and structural properties that distinguish it from its human homologues. In this study, we now describe the expression, kinetic characterisation and crystal structure determination of the LDH from Plasmodium berghei. This enzyme is seen to have a similar kinetic profile to its P. falciparum counterpart, exhibiting the characteristic lack of substrate inhibition that distinguishes plasmodial from human LDHs. The crystal structure of P. berghei lactate dehydrogenase (PbLDH) shows a very similar active site arrangement to the P. falciparum enzyme. In particular, an insertion of five amino acid residues in the active site loop creates an enlarged volume in the substrate binding site, and characteristic changes in the residues lining the NADH cofactor binding pocket result in displacement of the cofactor relative to its observed position in mammalian and all other LDH structures. These results imply the special features previously described for PfLDH may be shared across the Plasmodium genus, supporting the universal application of therapeutics targeting this enzyme.
Subject(s)
L-Lactate Dehydrogenase/chemistry , Plasmodium berghei/enzymology , Plasmodium falciparum/enzymology , Amino Acid Sequence , Animals , Cloning, Molecular , Crystallization , Crystallography, X-Ray , Humans , Kinetics , L-Lactate Dehydrogenase/genetics , L-Lactate Dehydrogenase/metabolism , Models, Animal , Models, Molecular , Molecular Sequence Data , Sequence Analysis, DNAABSTRACT
Lactate dehydrogenase (LDH) interconverts pyruvate and lactate with concomitant interconversion of NADH and NAD(+). Although crystal structures of a variety of LDH have previously been described, a notable absence has been any of the three known human forms of this glycolytic enzyme. We have now determined the crystal structures of two isoforms of human LDH-the M form, predominantly found in muscle; and the H form, found mainly in cardiac muscle. Both structures have been crystallized as ternary complexes in the presence of the NADH cofactor and oxamate, a substrate-like inhibitor. Although each of these isoforms has different kinetic properties, the domain structure, subunit association, and active-site regions are indistinguishable between the two structures. The pK(a) that governs the K(M) for pyruvate for the two isozymes is found to differ by about 0.94 pH units, consistent with variation in pK(a) of the active-site histidine. The close similarity of these crystal structures suggests the distinctive activity of these enzyme isoforms is likely to result directly from variation of charged surface residues peripheral to the active site, a hypothesis supported by electrostatic calculations based on each structure. Proteins 2001;43:175-185.
Subject(s)
Isoenzymes/chemistry , Isoenzymes/metabolism , L-Lactate Dehydrogenase/chemistry , L-Lactate Dehydrogenase/metabolism , Crystallization , Humans , Kinetics , Lactate Dehydrogenase 5 , Models, Molecular , Static Electricity , Structure-Activity RelationshipABSTRACT
ABSTRACT Short-chain acyl-CoA dehydrogenase (SCAD) deficiency is considered a rare inherited mitochondrial fatty acid oxidation disorder. Less than 10 patients have been reported, diagnosed on the basis of ethylmalonic aciduria and low SCAD activity in cultured fibroblast. However, mild ethylmalonic aciduria, a biochemical marker of functional SCAD deficiency in vivo, is a common finding in patients suspected of having metabolic disorders. Based on previous observations, we have proposed that ethylmalonic aciduria in a small proportion of cases is caused by pathogenic SCAD gene mutations, and SCAD deficiency can be demonstrated in fibroblasts. Another - much more frequent - group of patients with mild ethylmalonic aciduria has functional SCAD deficiency due to the presence of susceptibility SCAD gene variations, i.e. 625G>A and 511C>T, in whom a variable or moderately reduced SCAD activity in fibroblasts may still be clinically relevant. To substantiate this notion we performed sequence analysis of the SCAD gene in 10 patients with ethylmalonic aciduria and diagnosed with SCAD deficiency in fibroblasts. Surprisingly, only one of the 10 patients carried pathogenic mutations in both alleles, while five were double heterozygotes for a pathogenic mutation in one allele and the 625G>A susceptibility variation in the other. The remaining four patients carried only either the 511C>T or the 625G>A variations in each allele. Our findings document that patients carrying these SCAD gene variations may develop clinically relevant SCAD deficiency, and that patients with even mild ethylmalonic aciduria should be tested for these variations.
Subject(s)
Acyl-CoA Dehydrogenases/genetics , Genetic Variation , Lipid Metabolism, Inborn Errors/genetics , Acyl-CoA Dehydrogenase , DNA, Complementary , Female , Haplotypes , Humans , Infant , Infant, Newborn , Male , Mutagenesis, Site-Directed , MutationABSTRACT
Acute lung injury models demonstrate that high-frequency oscillatory ventilation (HFOV) improves lung function, mechanics, and histopathology with reduced inflammatory mediators. Neither human HFOV trials nor premature animal studies have adequately evaluated these factors during prolonged HFOV. The objective of this study was to compare the effect of prolonged HFOV with low tidal volume (VT) positive pressure ventilation (LV-PPV) in an immature baboon model for neonatal chronic lung disease (CLD). After administration of prenatal steroids, 18 baboons were delivered by cesarean section at 125 d (term = 185 d), treated with exogenous surfactant, then randomized to either HFOV or LV-PPV by 5 min age. Animals were maintained on oxygen on an "as needed" basis and on nutritional support for 1 to 2 mo. Serial pulmonary function testing (PFT) was performed. Tracheal aspirates were analyzed for interleukin-6 (IL-6), IL-8, tumor necrosis factor-alpha (TNF-alpha), IL-1beta, and IL-10. Lungs were inflation fixed for morphometric analyses. From 12 h through 10 d age, HFOV animals had consistently lower fraction of inspired oxygen (FI(O(2))) and higher a/ A ratio. Pulmonary mechanics were significantly improved in HFOV animals at nearly every time point analyzed from 12 h to 28 d. There were no consistent differences in tracheal IL-6, TNF-alpha, IL-1beta, or IL-10 after 24 h age. Higher tracheal IL-8 values and macrophage/monocyte numbers were found in LV-PPV animals after 1 wk and 3 to 4 wk ventilation. Both groups exhibited pulmonary pathologic lesions found in extremely immature humans, including alveolar hypoplasia, variable saccular wall fibrosis, and minimal airway disease. HFOV animals had significantly better lung inflation patterns by panel of standards analysis. Early, prolonged HFOV significantly improved early lung function with sustained improvement in pulmonary mechanics out to 28 d. Immature baboons managed with HFOV had less pulmonary inflammation in the hyaline membrane disease (HMD) recovery phase. Though enhanced alveolization was not observed, HFOV for 1 to 2 mo resulted in consistently more uniform lung inflation than LV-PPV.
Subject(s)
Cytokines/metabolism , High-Frequency Ventilation , Lung Diseases/etiology , Respiratory Mechanics , Trachea/metabolism , Animals , Animals, Newborn , Bronchopulmonary Dysplasia/etiology , Bronchopulmonary Dysplasia/pathology , Bronchopulmonary Dysplasia/physiopathology , Chronic Disease , Combined Modality Therapy , Female , Gestational Age , Humans , Infant, Newborn , Lung Diseases/metabolism , Lung Diseases/pathology , Lung Diseases/physiopathology , Male , Papio , Positive-Pressure Respiration , Pulmonary Surfactants/therapeutic use , Respiratory Distress Syndrome, Newborn/therapy , Tidal VolumeABSTRACT
Acyl-CoA dehydrogenase (ACAD) defects in isoleucine and valine catabolism have been proposed in clinically diverse patients with an abnormal pattern of metabolites in their urine, but they have not been proved enzymatically or genetically, and it is unknown whether one or two ACADs are involved. We investigated a patient with isolated 2-methylbutyrylglycinuria, suggestive of a defect in isoleucine catabolism. Enzyme assay of the patient's fibroblasts, using 2-methylbutyryl-CoA as substrate, confirmed the defect. Sequence analysis of candidate ACADs revealed heterozygosity for the common short-chain ACAD A625 variant allele and no mutations in ACAD-8 but a 100-bp deletion in short/branched-chain ACAD (SBCAD) cDNA from the patient. Our identification of the SBCAD gene structure (11 exons; >20 kb) enabled analysis of genomic DNA. This showed that the deletion was caused by skipping of exon 10, because of homozygosity for a 1228G-->A mutation in the patient. This mutation was not present in 118 control chromosomes. In vitro transcription/translation experiments and overexpression in COS cells confirmed the disease-causing nature of the mutant SBCAD protein and showed that ACAD-8 is an isobutyryl-CoA dehydrogenase and that both wild-type proteins are imported into mitochondria and form tetramers. In conclusion, we report the first mutation in the SBCAD gene, show that it results in an isolated defect in isoleucine catabolism, and indicate that ACAD-8 is a mitochondrial enzyme that functions in valine catabolism.
Subject(s)
Amino Acid Metabolism, Inborn Errors/enzymology , Amino Acid Metabolism, Inborn Errors/genetics , Isoleucine/metabolism , Oxidoreductases Acting on CH-CH Group Donors , Oxidoreductases/deficiency , Valine/metabolism , Alternative Splicing/genetics , Amino Acid Metabolism, Inborn Errors/metabolism , Amino Acid Sequence , Animals , Base Sequence , COS Cells , Child, Preschool , Consanguinity , DNA Mutational Analysis , Enzyme Stability , Exons , Female , Heterozygote , Humans , Introns , Male , Mitochondria/enzymology , Mitochondria/metabolism , Mutation/genetics , Oxidoreductases/genetics , Oxidoreductases/metabolism , Pakistan , Protein Transport , Sequence Deletion/genetics , TransfectionABSTRACT
Haemophilus influenzae penetrates between epithelial cells via an unknown mechanism. A chromosomal library of nonencapsulated H. influenzae strain A960053 DNA was constructed in Escherichia coli DH5alpha to identify bacterial genes contributing to this paracytosis. Two E. coli clones that contained open reading frames (ORFs) homologous to HI0636 to HI0641 of H. influenzae strain Rd and that showed an increased penetration in epithelial cell layers of the human bronchial epithelial cell line NCI-H292 were identified. ORFs HI0636 and HI0638, encoding two small proteins of unknown functions, were further investigated. The clone containing ORFs HI0636 and HI0637 as well as the clone containing ORF HI0638 showed a significant increase in penetration. Disruption of HI0638 by kanamycin box insertion in H. influenzae strain A960053 resulted in loss of penetration into the epithelial cell layers. Disruption of HI0636 had no effect on penetration in this model system. Since a role for HI0637 in the paracytosis of H. influenzae is very unlikely because it encodes TrpS, we conclude that the protein encoded by ORF HI0638 may function as a paracytin, while that encoded by HI0636 may have an auxiliary function.
Subject(s)
Genes, Bacterial , Haemophilus influenzae/genetics , Haemophilus influenzae/pathogenicity , Lung/microbiology , Respiratory Mucosa/microbiology , Bacterial Adhesion , Bacterial Typing Techniques , Epithelial Cells/microbiology , Escherichia coli/genetics , Genomic Library , Gentamicins/pharmacology , Haemophilus influenzae/classification , Haemophilus influenzae/drug effects , Humans , Mutagenesis , Open Reading Frames , Restriction Mapping , Sequence Analysis, DNA , Tumor Cells, CulturedABSTRACT
Screen-printed electrodes coated with the nafion layer have been investigated for cholinesterase biosensor design. The butyrylcholinesterase (ChE) from horse serum was immobilised onto the nafion layer by cross-linking with glutaraldehyde vapours. The biosensors obtained showed better long-term stability and lower working potential in comparison to those obtained with no nafion coating. The sensitivity of a biosensor toward organophosphate pesticides is not affected by the nafion coating. The detection limits were found to be 3.5x10(-7) M for trichlorfon and 1.5x10(-7) M for coumaphos.
ABSTRACT
A borderline viability model of bronchopulmonary dysplasia (BPD)/chronic lung disease of infancy (CLD) with pathophysiologic parameters consistent with those in extremely immature humans with BPD/CLD is described. After prenatal steroid treatment of pregnant dams, 12 premature baboons were delivered by cesarean-section at 125 d (term gestation, 185 d), treated with exogenous surfactant, and maintained on appropriate oxygen and positive pressure ventilation for at least 1 to 2 mo. In spite of appropriate oxygenation (median FI(O(2)) at 28 d = 0.32; range, 0.21 to 0.50) and ventilatory strategies to prevent volutrauma, the baboons exhibited pulmonary pathologic lesions known to occur in extremely immature humans of less than 1,000 g: alveolar hypoplasia, variable saccular wall fibrosis, and minimal, if any, airway disease. The CLD baboon lungs showed significantly decreased alveolization and internal surface area measurements when compared with term and term + 2-mo air-breathing controls. A decrease in capillary vasculature was evident by PECAM staining, accompanied by dysmorphic changes. Significant elevations of TNF-alpha, IL-6, IL-8 levels, but not of IL-1beta and IL-10, in tracheal aspirate fluids were present at various times during the period of ventilatory support, supporting a role for mediator-induced autoinflammation. IL-8 levels were elevated in necropsy lavages of animals with significant lung infection. This model demonstrates that impaired alveolization and capillary development occur in immature lungs, even in the absence of marked hyperoxia and high ventilation settings.
Subject(s)
Bronchopulmonary Dysplasia/pathology , Disease Models, Animal , Animals , Animals, Newborn , Bronchopulmonary Dysplasia/physiopathology , Bronchopulmonary Dysplasia/therapy , Cell Count , Gestational Age , Humans , Immunohistochemistry , Infant, Newborn , Interleukins/analysis , Lung/metabolism , Lung/pathology , Papio , Respiration, Artificial , Trachea/metabolism , Trachea/pathology , Tumor Necrosis Factor-alpha/analysisABSTRACT
A ratio-fluorescence assay was developed for on-line localization and quantification of lipid oxidation in living cells. The assay explores the oxidative sensitivity of C11-BODIPY(581/591). Upon oxidation, the fluorescence of this fluorophore shifts from red to green. The probe incorporates readily into cellular membranes and is about twice as sensitive to oxidation as arachidonic acid. Using confocal microscopy, the cumene hydroperoxide-induced oxidation of C11-BODIPY(581/591) was visualized at the sub-cellular level in rat-1 fibroblasts. Preloading of the cells with tocopherol retarded this oxidation. The data demonstrate that C11-BODIPY(581/591) is a valuable tool to quantify lipid oxidation and anti-oxidant efficacy in single cells.