New methods for the study of the carbon monoxide compounds of respiratory enzymes. 1953.
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Carbon Monoxide↗
Biomedical subjects
L Smith
Publications and source records attributed to L Smith.
Direct transesterification of plasma fatty acids for the diagnosis of essential fatty acid deficiency in cystic fibrosis.
This study was aimed at redefining criteria for essential fatty acid (EFA) deficiency with the use of the direct transesterification procedure (1986. J. Lipid Res. 27: 114-120) and at determining whether a simple assay of total fatty acids (FA) is as predictive of EFA deficiency as the FA pattern from plasma, red cell, and platelet phospholipids. Fasting blood samples were taken from 163 cystic fibrosis (CF) patients who were encouraged to consume 35-40% of their calories as fat. Their mean (+/- SD) age was 9.6 +/- 4.8 yr. The control group consisted of 44 unaffected siblings aged 13.1 +/- 3.1 yr. The 20:3(n-9)/20:4(n-6) ratio in 77 (47%) CF children was more than 2 SD above the values (mean +/- SD) of 0.021 +/- 0.007 obtained in the 44 controls. Groups of EFA-sufficient (n = 10) and EFA-deficient (n = 7) subjects were selected for further studies. The plasma total FA 20:3(n-9)/20:4(n-6) ratios of 0.029 +/- 0.003 in EFA-sufficient and of 0.216 +/- 0.103 in EFA-deficient was as good a discriminant as FA in phospholipids from plasma, red cell PC, and platelets. Among the 21 individual fatty acids, 20:3(n-9), which was also found in controls, and 16:1(n-7) (palmitoleic) proved to be the most sensitive indices of EFA deficiency. They are equally reliable in plasma, red cells, and platelets, but the inverse linear relationship (r = -0.91) between the n-7 family and 18:2(n-6) proved to be more closely associated with EFA deficiency than the one (r = 0.66) between 20:3(n-9) and 20:4(n-6).(ABSTRACT TRUNCATED AT 250 WORDS)
Adolescent↗
Proteolytic and lysosomal enzymes in acute trauma-induced lung injury in sheep.
Surgical preparation of a lung lymph fistula in sheep was previously shown to induce temporary permeability disturbance of pulmonary microvessels. Flow and composition of lung lymph were compared in 19 sheep with acute, and 16 with chronic lymph fistula. Lymph and plasma were analyzed regarding activities of prothrombin, antithrombin III, prekallikrein, kallikrein, functional kallikrein inhibitors and 11 acid hydrolases of lysosomal origin. Levels of prothrombin and antithrombin III fell postoperatively in plasma and reached even lower values in lymph, indicating systemic activation of the coagulation system as well as local activation in the lung. The studied components of the plasma-kallikrein system were unchanged. Lysosomal enzymes were released in lung lymph, reflecting cell injury in the lung. Local activation of proteolytic enzyme systems thus occurs in the lung after trauma. Analysis of organ-specific lymph (= interstitial fluid) may contribute to better understanding of the pathophysiology of trauma.
Animals↗
Histopathologic characteristics and ultrastructure of aging skin.
Histologic and ultrastructural examination of skin biopsy specimens from younger compared to older persons documents age-related structural alteration in the epidermis, dermal-epidermal junction, dermis, and the epidermal appendages including hair follicles, sebaceous glands, and sweat ducts and glands. The fine, regular epidermal surface patterns change to coarser and less regular ridges with aging. Epidermal projections into the dermis are retracted and the dermal-epidermal junction is flattened. The dermis becomes thinner; there is less fibrous collagen and less elastic fiber in older skin, but the elastic component may appear increased compared to collagen. Elastic fibers may become frayed, porous, and matted together. The density of blood vessels is reduced and, in particular, there are fewer capillary loops in the papillary dermis. There are fewer as well as structurally altered hair follicles, sebaceous glands, and sweat glands with increasing age.
Adult↗
Relationship of decreased hepatic lipase activity and lipoprotein abnormalities to essential fatty acid deficiency in cystic fibrosis patients.
Polyunsaturated fatty acids are known to affect plasma lipids and lipoproteins but there is no information on the effect of essential fatty acid (EFA) deficiency on lipoprotein composition. The purpose of this study was to characterize lipoproteins from 17 cystic fibrosis (CF) patients in relationship to their EFA status (eicosatrienoic/arachidonic acid ratio) and compare them with those of 10 healthy siblings (SIB) and of 10 unrelated controls. In 7 EFA-deficient (EFAD) and 10 EFA-sufficient (EFAS) patients, hypocholesterolemia was associated with a decrease of HDL-cholesterol and of LDL-cholesterol which was more marked in the EFAD group. Similarly, although triglyceride enrichment of VLDL, LDL, HDL2, and HDL3 with a concomitant reduction of cholesteryl esters from all particles except HDL2 was observed in both CF groups, it was more sizable in the EFAD patients. These changes led to an increase in the particle size of VLDL, LDL, and HDL2 whereas the distribution of HDL3 was skewed to smaller particles. Alterations in the apoprotein composition of particles were greater in EFAD than in EFAS. A decrease of total postheparin lipolytic activity was observed in the two groups of CF patients as well as in siblings. It was entirely accounted for by hepatic lipase (mumol FFA/ml per h) which was more severely diminished in EFAD (2.8 +/- 0.6) than in EFAS (4.4 +/- 0.7) and SIB (5.1 +/- 0.5). Although the two groups of CF children differed in terms of growth, severity of malabsorption, and vitamin E status, these data suggest that disturbance of lipoprotein concentration, composition, size, and metabolism (hepatic lipase) may be in part related to EFA deficiency. Further studies are necessary to explore the effect of EFA deficiency on hepatic lipase activity.
Adolescent↗
The battle over health insurance.
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Data Collection↗
Screening and rescreening for colorectal cancer. A controlled trial of fecal occult blood testing in 27,700 subjects.
All inhabitants of the city of Göteborg who in 1982 were between 60 and 64 years of age (27,700) were randomly divided into a test and a control group. The 13,759 subjects in the test group were invited to perform Hemoccult II (Smith Kline Diagnostic, Sunnyvale, CA) fecal occult blood testing over 3 days and to repeat the testing after 16 to 22 months. At the first screening 9,040 (66%) completed the test, and 7,770 (58%) completed the test at the second screening. In the first screening the test group was divided into two subgroups in which the tests were rehydrated and unhydrated before development. All tests were rehydrated in the second screening; 1.9% and 5.8% of the tests were positive in the unhydrated and rehydrated subgroups, respectively. The number of diagnosed neoplasms in the first screening was significantly larger (P less than 0.01) in the rehydrated group compared to the unhydrated group, 50 and 24 neoplasms, respectively. Sixteen of 61 carcinomas in the test group were found in the interval between the two screenings, 19 of the carcinomas at the second screening, and ten among the nonresponders. Rehydration of the Hemoccult II test is a necessity. Significantly more carcinomas (61) were found in the test group compared to the control group (20). There was a trend toward favorable tumor staging in the test group compared to the control group.
Clinical Trials as Topic↗
Investigation of endosomal compartments involved in endocytosis and transcytosis of polymeric immunoglobulin A by subcellular fractionation of perfused isolated rat liver.
1. A gamma camera was used to monitor continuously the uptake of radiolabelled polymeric immunoglobulin A (pIgA) into the rat body after intravenous injection. Uptake into liver was fast but, since the peak of liver labelling occurred only after 9-15 min, it was not sufficiently rapid to constitute a pulse dose. A perfused, isolated rat liver system was therefore established which could be given a single pass dose of pIgA; a variety of tests showed such livers remained viable for at least 3 h and could be subsequently fractionated on Ficoll and Nycodenz gradients with normal distributions of marker enzymes. 2. Subcellular fractionation at different times after a single pass dose of pIgA showed that whilst pIgA appeared sequentially in sinusoidal plasma membrane, light endosomes, dense endosomes, very dense endosomes and lysosomes as in vivo, the predominance of pIgA in the light endosome compartment disappeared much earlier than after injection in vivo of pIgA, presumably because this compartment was not being continuously loaded over the first 10-15 min. The time course of appearance of label in bile was unchanged. A large excess of unlabelled asialofetuin did not change these patterns, indicating that the asialoglycoprotein receptor was not involved. 3. Low doses of the microtubule agent colchicine reduced the proportion of pIgA reaching the bile, but subcellular fractionation of treated liver showed that distribution of label amongst liver fractions was little changed, although the overall liver pIgA content had increased. This would suggest that pIgA did not remain in the common compartment which could have supplied bile or lysosomes but rather flowed out of it as rapidly as in untreated liver but towards those compartments supplying the lysosomes. 4. Experiments with nocodazole, which reversibly disrupts microtubules, showed that very little of the pIgA taken into an inhibited liver appeared in the bile after nocodazole was removed 30 min later, even though a second dose of pIgA, given after nocodazole removal, appeared in bile with a normal time course. The first dose of pIgA must therefore have passed beyond the compartments competent to supply the bile before nocodazole was removed. Such compartments were undamaged since the second dose of pIgA appeared in bile normally. We therefore conclude that the bulk of pIgA must be supplied to the bile from light or dense endosomes rather than from very dense endosomes and lysosomes.
Animals↗
Kinetics of the interaction of the cytochrome c oxidase of Paracoccus denitrificans with its own and bovine cytochrome c.
We have devised a relatively simple method for the purification of cytochrome aa3 of Paracoccus denitrificans with three major subunits similar to those of the larger subunits of the mitochondrial cytochrome oxidase. This preparation has no c-type cytochrome. Studies were made of the oxidation of soluble cytochromes c from bovine heart and Paracoccus. The cytochrome-c oxidase activity was stimulated by low concentrations of either cytochrome c, providing an explanation for the multiphasic nature of plots of v/S versus v. Kinetics of the oxidation of bovine cytochrome c by the Paracoccus oxidase resembled those of bovine oxidase with bovine cytochrome c in every way; the Paracoccus oxidase with bovine cytochrome c can serve as an appropriate model for the mitochondrial system. The kinetics of the oxidation of the soluble Paracoccus cytochrome c by the Paracoccus oxidase were different from those seen with bovine cytochrome c, but resembled the latter if poly(L-lysine) was added to the assays. The important difference between the two species of cytochrome c is the more highly negative hemisphere on the side of the molecule way from the heme crevice in the Paracoccus cytochrome. Thus, the data emphasize the importance of all of the charged groups on cytochrome c in influencing the binding or electron transfer reactions of this oxidation-reduction system. The data also permit some interesting connotations about the possible evolution from the bacterial to the mitochondrial electron transport system.
Animals↗
Role of IIb-IIIa-like glycoproteins in cell-substratum adhesion of human melanoma cells.
The platelet fibrinogen receptor, glycoprotein complex IIb-IIIa, was isolated from human platelets by lectin and monoclonal antibody affinity chromatography and a polyclonal antiserum (anti-IIb-IIIa) was generated and used to probe for the presence and function of IIb-IIIa-like molecules in two adherent human cell lines. Both C32 melanoma cells and WI38 fibroblasts expressed a IIb-IIIa-like complex on their surface as indicated by immunoprecipitation of detergent extracts of surface radiolabeled cells. When added to cells plated in medium containing 10% serum, the anti-IIb-IIIa antiserum perturbed the adhesion of C32 melanoma cells, but not of WI38 fibroblasts. In a serum-free system, anti-IIb-IIIa antibodies inhibited attachment and spreading of C32 cells to fibrinogen, vitronectin, and fibronectin adsorbed to glass. Anti-IIb-IIIa had no effect on the attachment and spreading of WI38 cells to the extracellular matrix proteins, however. Thus, the IIb-IIIa-like complex appears to play a predominant role in cell-substratum adhesion of C32 cells, but not WI38 cells, and may result from the fact that, on a protein basis, the C32 melanoma cells express approximately 3 times more complex on their surface than do WI38 fibroblasts. The results suggest that the relative abundance of a particular adhesion receptor on the cell surface may govern its importance to cell-substratum adhesion.
Cell Adhesion↗
Pairing alters the effect of progesterone on nesting in female C57BL/10Sn mice.
Progesterone treatments have been shown to increase nesting levels of isolated female mice. Here we compared the effect of exogenous progesterone on the nesting behavior of C57BL/10Sn female Mus domesticus housed individually to nesting levels of the same mice housed in pairs. Progesterone increased nesting by isolated females but had no significant effect on the nesting scores of the same mice when they were grouped into pairs. The effect of exogenous progesterone on nesting levels in C57BL/10Sn females appears to depend on social circumstances.
Animals↗
Amplitude compression and profound hearing loss.
Nine subjects with prelingually acquired, sensorineural, hearing loss were given a three-interval, forced-choice, test of speech pattern contrast perception under two amplification conditions. The first involved adjustment of the low and high frequency outputs of a two-channel Master Hearing Aid to each subject's highest comfortable level, but without compression of the short-term dynamic range of the signal. The second involved the additional compression of a 30 dB input range into the subject's dynamic range of hearing, as measured by the difference between speech awareness threshold and highest comfortable level, in each of the two channels. One of the subjects performed much better with compression than without. Among the other eight, however, there was a small but significant reduction of performance when compression was introduced. It is proposed that the one positive result is due to the increased audibility of speech cues made possible by amplitude compression. It is further proposed that the negative results are due mainly to the distortions of time-intensity cues introduced by amplitude compression. The results suggest that, in terms of potential access to meaningful speech cues, the addition of amplitude compression, to an otherwise optimized signal, is unnecessary, or even detrimental, for most profoundly deaf subjects, but could be beneficial for some.
Adolescent↗
Digestive and absorptive phase anomalies associated with the exocrine pancreatic insufficiency of cystic fibrosis.
The pancreas has an enormous reserve capacity, and significant malabsorption usually signals complete absence of exocrine function. However, there is evidence that acid lipases of nonpancreatic origin play an important compensatory role. Complete duodenal hydrolysis of fat requires a series of complex interdependent physicochemical events involving pancreatic lipase, colipase, phospholipase A2, and bile salts in an environment where the pH must be close to neutrality. Lipolytic products must then be shuttled through the unstirred water layer to the surface of the microvillus membrane by ionized bile salts, which must be present in sufficient concentrations to form micelles. In pancreatic insufficiency, there is not only a defective lipolytic phase but also an impaired micellar phase. The output of bile salts is decreased because of increased fecal loss. Furthermore, a significant percentage of bile salts precipitate because the duodenum is acidic and there is a large predominance of glycine conjugates. Although much less work has been done on the absorptive phase of patients with pancreatic insufficiency, there is tentative evidence that defective phospholipid absorption, essential fatty acid deficiency, and protein malnutrition could impair the absorptive phase, particularly chylomicron formation. Although significant advances have been made in our understanding of factors responsible for malabsorption associated with pancreatic insufficiency, much remains to be done for the further delineation of defects. It is hoped that this will lead to further refinements of enzyme preparations and to new strategies of intervention.
Cystic Fibrosis↗
A microelectrometric titration method for measurement of total intracellular Cl- concentration.
A microelectrometric titration method is described to measure picomole amounts of Cl- present in solutions at micromolar concentrations. This method was used to measure total intracellular chloride concentration ([Cl-]i) in leukocytes. Through the use of submicroliter samples, [Cl-] can be measured in the range of 5-500 microM. There is no measurable interference from other ions normally present in the cell and no intracellular ion is falsely measured as Cl-. [Cl-]i determined by the conventional coulometric titration and the new microelectrometric titration method was the same. Among the commonly used substituting anions, thiocyanate was the only one falsely measured as Cl- and should be avoided in experiments using this method. Because only picomole amounts of Cl- are required, measurements of intracellular pH and total concentrations of other intracellular ions can be done in the same lysate of as few as 5 X 10(5) cells. This feature should make it possible to study the time course of changes in [Cl-]i together with measurements of other intracellular ions following various physiological or experimental maneuvers.
Animals↗
Effect of increased hydrostatic pressure on lymphatic elimination of hyaluronan from sheep lung.
The effects of increased hydrostatic pressure on the concentrations of hyaluronan (hyaluronic acid) in lung lymph and serum were investigated in awake sheep with a cannula in the efferent vessel from the caudal mediastinal lymph node. Lung lymph was sampled at base line [left atrial pressure (LAP) 6.5 +/- 1.7 mmHg] and after two increases of LAP to 25.7 +/- 2.2 mmHg (level 1) and 37.0 +/- 5.1 mmHg (level 2). The lung lymph flow increased from 1.9 +/- 0.5 at base line to 9.3 +/- 2.2 and 15.9 +/- 0.7 ml/30 min, and the lymph-to-plasma concentration ratio of total protein decreased from 0.63 +/- 0.02 to 0.32 +/- 0.04 and 0.32 +/- 0.05 at the two elevated levels of LAP, respectively. The hyaluronan concentration in lung lymph was unchanged, and there was a flow-dependent elimination of hyaluronan from the lung that increased from 23 +/- 8 to 87 +/- 19 and 137 +/- 37 micrograms/30 min, respectively. The lung concentration of hyaluronan was 167 +/- 28 micrograms/g fresh lung, and at base line it was calculated that slightly less than 2% of the lung hyaluronan was eliminated by the lymphatic route in 24 h. If extrapolated to 24 h, the elimination rate of hyaluronan seen during elevated LAP would result in lymphatic elimination of 18% of the lung hyaluronan over this time period. Since hyaluronan is responsible for part of the protein exclusion in the extracellular matrix, it is plausible that washout of interstitial hyaluronan contributes to the decrease in albumin exclusion from the interstitium that occurs after an elevation of LAP.
Animals↗
Local pulmonary activation of proteolytic enzymes after Escherichia-coli-induced lung injury in sheep.
Activation of several cascade systems, e.g. coagulation, fibrinolysis, kallikreinkinin, complement and eicosanoid systems, has been implicated in the etiology of septic-lung microvascular injury. A chronic lung lymph fistula preparation in sheep (n = 9) was used to study coagulation, kallikrein-kinin and eicosanoids during Escherichia coli septicemia. Lung lymph flow and lymph composition indicated an increased lung microvascular permeability approximately 2 h after infusion of bacteria. Stable prothrombin and antithrombin III levels in lymph contradicted local activation of the coagulation cascade in the lung and systemic activation was not evident until 4 h after bacteria infusion. Lymph thromboxane B2 and 6-keto PGF1 alpha peaked early (1 h). Reduced lymph prekallikrein and kallikrein inhibitors indicated local activation of this system in the lung. Systemic activation of kallikrein could not be demonstrated. Thus, (1) changes in systemic blood may not adequately reflect local events and (2) studies of proteolytic enzymes and other inflammatory mediators in lung may contribute to clarifying the etiology of microvascular injury.
Animals↗
Gallium scintigraphy in small cell lung cancer.
We have undertaken gallium imaging studies in 49 patients with histologically proven small cell lung cancer. Tracer uptake in the primary tumour was seen in 98% of cases. Twenty five patients underwent repeat scanning after induction chemotherapy and a correlation was demonstrated between conventional parameters of response and gallium scan changes (P less than 0.01). There was no correlation between initial gallium activity and subsequent chemoresponse (which was evaluated in 32 patients) or survival (measured in 42 patients). Ten patients who had shown a complete response to induction treatment were followed up with gallium scans repeated at three monthly intervals. Such longitudinal studies were particularly helpful in excluding tumour activity when the appearance of the chest radiographs were difficult to interpret.
Antineoplastic Agents↗
Kinetics of the interaction of cytochrome c oxidase of Paracoccus denitrificans with Paracoccus and mitochondrial cytochrome c.
We have studied the reactions of the oxidase of Paracoccus dentrificans with its membrane-bound cytochrome c and with soluble cytochrome c550 of Paracoccus and of bovine heart. The turnover rate of Paracoccus oxidase with membrane-bound cytochrome c is high, approaching 1000/sec. at 25 degrees. When soluble cytochrome c is added to the electron transport chain oxidizing NADH or succinate, no increase in 02 uptake is observed. When the oxidase is reacting with the membrane-bound cytochrome c, the reaction site is not exposed for reaction with soluble cytochrome c. We have purified the Paracoccus oxidase, following relatively simple methodology. It has three major subunits similar in molecular weight to those of the larger subunits of the bovine oxidase. When reconstituted in the presence of asolectin, it is just as active as the intact membrane-bound oxidase in reaction with soluble cytochrome c. The soluble cytochrome c reacts directly with the cytochrome aa3. We found direct evidence that the oxidase is stimulated in the presence of low concentrations of cytochrome c. The stimulatory effect could be the explanation for the so-called "high affinity" site for reaction with cytochrome c. The reaction of bovine cytochrome c with Paracoccus oxidase resembles that with the bovine oxidase in every way tested. The Paracoccus oxidase must have a cytochrome c binding site equivalent to that of the bovine enzyme. The reaction of the Paracoccus oxidase with its own soluble cytochrome c550, which has a highly negative hemisphere on the side of the molecule away from the heme crevice, has different properties from those seen in its reaction with bovine cytochrome c. However the properties all change to be like those with bovine cytochrome c on addition of poly-L-lysine. These data emphasize the importance of all of the charged groups on the cytochrome c in influencing binding or electron transfer reactions. The respiratory chain on the membranes of a cytochrome c-deficient mutant can reduce cytochrome aa3 using NADH as substrate in a manner similar to that of the wild type, although at somewhat lower rate, suggesting diffusional encounter of the large complexes within the membrane. Our data permit speculations about the possible evolution from the bacterial to the mitochondrial electron transport system.
Animals↗