Automated analysis of the pyridinium crosslinks of collagen in tissue and urine using solid-phase extraction and reversed-phase high-performance liquid chromatography. A fully automated method for assaying the collagen crosslinking amino acids, pyridinoline and deoxypyridinoline, in human urine samples or tissue hydrolysates is described. Samples were processed using a Gilson ASPEC system with solid-phase extraction of the crosslinks on columns containing 100 mg of microgranular cellulose. Introduction of an additional solvent step during sample preparation allowed direct analysis by reversed-phase HPLC and elimination of the drying step used previously in a manual method. Use of a synthetic pyridinoline derivative as internal standard enabled accurate quantification of the crosslinks by correcting for recoveries through the whole assay. Samples were analyzed in sequential mode with a total assay time of 30 min. The automated assay showed close correlation with the manual method for both free and total crosslink determinations in human urine (r > 0.97). Reproducibility was improved, as seen from replicate analyses of human urine (CV < 3% for automated pyridinoline measurement compared with 8-12% previously observed for the manual method). Crosslink excretion is the most useful marker of collagen degradation in metabolic bone diseases and arthritic disorders. The automated assay which has been developed is rapid, convenient, and reliable and will greatly facilitate the monitoring of urinary collagen crosslinks and their tissue levels in clinical investigations.


Carbohydrate analysis of water-soluble uronic acid-containing polysaccharides with high-performance anion-exchange chromatography using methanolysis combined with TFA hydrolysis is superior to four other methods. Sulfuric acid hydrolysis according to the Saeman procedure, TFA hydrolysis, and methanolysis combined with TFA hydrolysis were compared for the hydrolysis of water-soluble uronic acid-containing polysaccharides originating from fungi, plants, and animals. The constituent sugar residues released were subsequently analyzed by either conventional GLC analysis of alditol acetates or high-performance anion-exchange chromatography with pulsed-amperometric detection. It was shown that TFA hydrolysis alone is not sufficient for complete hydrolysis. Sulfuric acid hydrolysis of these polysaccharides resulted in low recoveries of 6-deoxy-sugar residues. Best results were obtained by methanolysis combined with TFA hydrolysis. Methanolysis with 2 M HCl prior to TFA hydrolysis resulted in complete liberation of monosaccharides from pectic material and from most fungal and animal polysaccharides tested. Any incomplete hydrolysis could be assessed easily by HPAEC, by the detection of characteristic oligomeric products, which is difficult using alternative methods currently in use. Methanolysis followed by TFA hydrolysis of 20 micrograms water-soluble uronic acid containing polysaccharides and subsequent analysis of the liberated sugar residues by HPAEC allowed us to determine the carbohydrate composition of these polysaccharides rapidly and accurately in one assay without the need for derivatization.


Fluorimetric DNA assay for cell growth estimation. A growth assay using the fluorescent dye Hoechst 33258 has been developed which allows sensitive and rapid analysis of the DNA content of a variety of cell types including keratinocytes and mucus-secreting cells, and which requires a minimum of liquid handling. The assay can detect as few as 500 diploid human cells, and is compatible with the simultaneous detection of [3H]thymidine incorporation in the same cultures.


Microsequence analysis of electroblotted proteins. II. Comparison of sequence performance on different types of PVDF membranes. The influence of different types of polyvinylidene difluoride (PVDF) membranes on gas phase sequence performance has been evaluated. These PVDF membranes have been classified as either high retention (Trans-Blot and ProBlott) or low retention membranes (Immobilon-P) based on their ability to bind proteins during electroblotting from gels. Initial yields, repetitive yields, and extraction efficiency of the anilinothiazolinone amino acid derivatives have been compared for several standard proteins that have been either electroblotted or loaded onto PVDF membranes by direct adsorption. These results show that the major differences in initial sequence yields between membranes arise from differences in the amount of protein actually transferred to the membrane rather than sequencer-related factors. In contrast to several previous observations from other laboratories, more tightly bound proteins do not sequence with lower initial yields and initial yields are not affected by the ratio of surface area to protein. The stronger binding on high retention PVDF membranes does not adversely affect recoveries of difficult to extract, or very hydrophobic, amino acid derivatives. Several amino acids, especially tryptophan, are actually recovered in dramatically higher yield on high retention membranes compared with either Immobilon or glass filters. At the same time, the protein and peptide binding properties of high retention membranes will frequently improve the repetitive yield by minimizing sample extraction during the sequencer cycle. Stronger protein binding together with improved electroblotting yields offer substantially improved sequence performance when high retention PVDF membranes are used.


Quantitative immunoassay of human deoxycytidine kinase in malignant cells. Deoxycytidine kinase (dCK) is necessary for the activity of several nucleosides used for the chemotherapy of cancer and AIDS. However, the measurement of dCK catalytic activity in crude cell extracts may be imprecise, due to the presence of phosphatases and nucleotidases that degrade the enzyme products. We describe a simple immunoassay for dCK that can measure accurately as little as 5 ng enzyme protein in crude tissue extracts. The assay enabled us to show (i) that mutant cells deficient in dCK activity lack immunoreactive dCK protein, (ii) that dCK catalytic activity and immunoreactivity correlate closely in human tumors, and (iii) that immunoreactive dCK is particularly high in lymphocytes and lymphoid malignancies, although certain solid tumors may also contain the enzyme. The immunoassay of dCK could prove useful in the selection and monitoring of patients who are being treated with nucleosides that are activated by this enzyme.


Purification of cloned and genomic DNA by guanidine thiocyanate/isobutyl alcohol fractionation. We have developed a rapid and efficient method to purify cloned or high molecular weight eukaryotic DNA from cell lysates using guanidine thiocyanate and isobutyl alcohol. This simple two-step extraction procedure utilizes isobutyl alcohol to fractionate DNA away from cell lysates containing guanidine thiocyanate, yielding a relatively large quantity of pure DNA. The yield of DNA is approximately twice that obtained by the organic extraction method. DNA purified with this chaotropic-based protocol is suitable for a variety of applications including restriction analysis, double-stranded sequencing, PCR, subcloning, and transcription.


Multiple nucleic acid labeling and rainbow detection. A method which allows discrete nucleic acid sequences to be detected with differently colored hybridization signals on the same blot involving only a single hybridization step is described. Nucleic acid probes labeled with digoxigenin, fluorescein, or biotin are hybridized simultaneously to immobilized target nucleic acids. Differential colorimetric detection is carried out in consecutive alkaline phosphatase-based immunoassays with one of three 3-hydroxy-2-naphthoic acid anilide phosphate/diazonium salt combinations as substrate. Each label is visualized by a different color precipitate (green, red, and blue) directly on the membrane. We demonstrate the use of this method in multicolor plasmid mapping, detection of different genomic sequences on a single Southern blot, discrimination of transcription levels in a Northern blot, and colony screening. Advantages and limitations of the method, as well as further applications, are discussed.


Characterization of immune complex components by dot blot analysis. A method is described for the characterization of immune complex components by dot blot analysis. After isolation by chromatographic techniques and precipitation with polyethylene glycol, immune complexes were dissociated in 0.1 M phosphate (pH 2) and bound to a nitrocellulose membrane in a dot blot unit. Biotinylated probes were then used to identify the following immune complex components: specific antigens, biologically active antibodies, antibody isotypes, antibody subclasses, antibody idiotypes, and rheumatoid factors. This nonradioactive procedure takes less than 2 h to perform and has been used to analyze immune complexes isolated from sera (rabbit and human) and synovial fluid (human).


Effect of subunit interactions on enzymatic activity of glutathione S-transferases: a radiation inactivation study. The glutathione S-transferases are a family of dimeric enzymes. Three isozymes from the alpha family, termed YaYa, YaYc, and YcYc, and three from the mu family, termed Yb1Yb1, Yb1Yb2, and Yb2Yb2, were purified from rat liver. Binding studies were performed by equilibrium dialysis using a radiolabeled product, S(-)[14C](dinitrophenyl)glutathione. Each isozyme contained two independent binding sites which had equal affinity for the ligand. The presence of two independent active sites per enzyme dimer suggests that each subunit contains a complete active site. This conclusion was examined further using radiation inactivation which also allowed for assessment of the importance of subunit interactions in catalytic activity. The activity target size of YaYa (47 kDa) was significantly larger than the protein monomer target size (31 kDa); similarly the activity target size of YaYc was that of the dimer (54 kDa). In contrast, the activity target sizes of Yb1Yb1 and Yb2Yb2 were the same, being 35 and 29 kDa, respectively, and the protein monomer target size of Yb1Yb1 also was similar, being 32 kDa. These data indicate that interactions between subunits are critical for the maintenance of enzymatic activity of alpha class enzymes whereas each subunit of the two mu class proteins is capable of independent catalytic activity.


Determination of radioligand specific activity using competition binding assays. Radioligand binding assays are routinely utilized in laboratories throughout the world to study receptors and their related binding sites, carrier proteins, and enzymes. To accurately estimate equilibrium binding parameters, such as the equilibrium dissociation constant (Kd) and maximal number of binding sites (Bmax), the investigator must know the correct value of the specific activity of the radioligand. If the specific activity is overestimated the Kd and Bmax values will be underestimated, while underestimation of the specific activity results in an overestimation of the Kd and Bmax. The present communication describes a simple and rapid method for determining the specific activity of a radioligand using homologous competition binding assays. Performing the competition assays at two or more different concentrations of the radioligand allows the specific activity to be determined from the IC50 values without the need of analytical methods to quantify minute amounts of the radioligand. In addition to providing the specific activity, use of this method estimates the Kd for the radioligand. This method was utilized to determine the specific activity and Kd for two blockers of the dopamine uptake carrier, [3H]GBR-12935 and [3H]-CFT, which share a common binding site in the striatum.


A fast and versatile method for extraction and quantitation of long-chain acyl-CoA esters from tissue: content of individual long-chain acyl-CoA esters in various tissues from fed rat. A method for the extraction of acyl-CoA esters from tissue, and their subsequent analysis by HPLC is described. The lipids are removed by a two-phase extraction in a chloroform/methanol/water system. The long-chain acyl-CoA esters are extracted using methanol and a high salt concentration (2 M ammonium acetate). Reextraction of the dry residue after evaporation of extraction solvent results in low overall recoveries (20%). By adding 1 mg/ml acyl-CoA-binding protein to the extraction solvent the overall recovery was increased to 55%. The method is easy and fast to perform and is thereby suitable for analysis of a large number of samples. The advantages of the method over previously published methods are discussed.


The use of a spectrophotometric assay to study the interaction of S-adenosylmethionine synthetase with methionine analogues. We have developed a continuous spectrophotometric assay for S-adenosylmethionine synthetase and, using this assay, have examined the interaction of five potential inhibitors with the E. coli enzyme. S-Vinylhomocysteine and S-allylhomocysteine were found to be substrates, while S-(methanethio)cysteine and S-(methanethio)homocysteine were found to be competitive inhibitors. S-Cyanohomocysteine is neither a substrate nor an inhibitor.


Regeneration of functional hemoglobin from iron(III) hemoglobin by reduction with hydrogen and a heterogeneous catalyst. Functional hemoglobin was regenerated from partially autoxidized hemoglobin by reduction with molecular hydrogen in the presence of a heterogeneous catalyst consisting of elemental platinum embedded in an electroactive polymer. The visible spectrum of the regenerated hemoglobin was identical to that of native iron(II) hemoglobin. The regenerated hemoglobin displayed highly cooperative oxygen-binding characteristics. P50 values for oxidized-regenerated hemoglobin samples were not different from native hemoglobin. The Hill coefficients for regenerated hemoglobin were slightly lower than the controls, possibly because of small amounts of irreversibly oxidized hemoglobin arising during the initial autoxidation. The advantages of the reduction system include: (1) the heterogeneous catalyst avoids the problem of protein adsorption onto bare platinum, (2) catalyst and reducing agent are easily removed from the protein, and (3) the by-product H+ is buffered easily.


A temperature-controlled, anaerobic cell for direct electrochemical studies. The construction and operation of a cell for temperature-controlled, direct electrochemical studies of oxygen-sensitive materials are described. The borosilicate cell contains a pyrolytic graphite working electrode, a Ag/AgCl reference electrode, and a platinum counter electrode, all of which can be readily interchanged with other types of electrodes. It is surrounded by a water jacket constructed of steel and Lexan, which can easily maintain temperatures between 4 degrees C and at least 90 degrees C. The entire cell was designed to minimize the number, complexity, and expense of components, as well as minimize required sample volume (250 microliters) and sources of oxygen leakage. As examples of the cell's utility, the redox properties of two common organic redox dyes, methyl viologen and thionin, were determined by differential pulse voltammetry at temperatures from 30 to 90 degrees C.


[The microvascularization of the penis of the steer (Bos taurus)]. The blood supply and the microvascularization of the bovine penis (Bos taurus) were demonstrated using the scanning electron microscope. The Corpus cavernosum penis of the bull consists of a surprisingly well-developed mesh of small to intermediate vessels. The architecture of the cavernous body is largely determined by trabeculae of connective tissue, the extent of which varies between penis segments. The Corpus cavernosum is arranged primarily as a ring around the trabeculae of connective tissue. Chambers of large bore are found in the central portions of the Corpus cavernosum penis, whereas a relatively fine vascular network predominates in the periphery. Vessels in the cavernous body of the urethra, on the other hand, show strictly parallel orientation to the urethra, which they surround like a sleeve. They also stand in close association with vessels in the outer layer of the Tunica albuginea. The vascular systems of the Corpus cavernosum penis and the Corpus spongiosum penis are not connected with one another. The vascular architecture of the Glans penis is characterized by the inclusion of well-developed vascular arcades at regular intervals. These course through a connective tissue matrix rich in glycoproteins and stretch to just under the skin of the penis, where they are intimately associated with a subpapillary network of arteries and veins. The venous legs of the vascular arcades are supplied by this network of veins. The Glans penis of the bull is considered to represent a specialization of the penile integument.


[Prenatal development of the horse ovary]. To answer the many open questions concerning the development of the horse's ovary, first the prenatal development was investigated. It resulted that follicles derive from the germinal epithelium and its cords, whereas the Leydig cells and the rete blastema originate from the mesonephros. In the second third of pregnancy the Leydig cells undergo an enormous proliferation, in the last third they degenerate. However this degeneration is not connected with the postnatal development of the ovulation groove.


[Comparative histological study of the reproductive system of the female llama (Lama guanicoe glama). I. Ovary]. In the present study a cytological, histological and morphometrical comparison between the ovaries of the llama, the cow and the sheep is presented, at two phases of the ovarian cycle. There were found differences in the amount of primordial and primary follicles, the size of secondary follicles and follicular cells, and type and distribution of the connective tissue inside the stroma of the ovary. It would be necessary to study the fine structure of the ovary and the so-called "embryological remnants", for its permanent appearance in most (50%) of the ovaries.


[The development of the yolk sac in ruminants (sheep and cattle)]. Yolk sac development was investigated in 69 ovine and 10 bovine embryos from the blastocyst stage to the 7th week of gestation. Light and electron microscopical findings are reported. The yolk sac in sheep and cattle is composed of an enlarged sac-like portion lying below the embryo and two ends which follow the elongated course of the trophoblast. In sheep, an open connection exists between the intestines and the yolk sac up to a crown-rump length (CRL) of 9 mm. It is closed by 12 mm CRL. The wall of the yolk sac is especially well vascularized in the enlarged, sac-like portion. Primary erythropoiesis occurs within the blood capillaries. In the blastocyst, the yolk sac entoderm is made up of elongated, flat cells. It becomes cuboidal in the 3 mm embryo (ovine) and later columnar. The up to 20 microns tall cells stain darkly and contain numerous light-colored vesicles. At 4.5 mm CRL light cells appear between the dark ones. Both cells are rich in rough endoplasmic reticulum (rER). The increased staining of the darker cells is due to an osmophilic cytoplasm and numerous, often parallel lamella of rER. The rER of the light cells is enlarged to irregularly-shaped cisternae, which nearly fill the entire cytoplasm and give them a rounded appearance. The dark cells contain polygonal nuclei, whereas those in the light cells are round with one or two nucleoli. The oval mitochondria have only a few peripheral cristae. Golgi fields are not very common. Cells of the entoderm are connected to one another over zonulae occludentes. They possess microvilli on the luminal surface and are supported by a basement membrane. From 5 mm CRL onwards (ovine), the yolk sac entoderm folds itself between the capillaries, thereby becoming stratified. The intercellular space between the cells expands as projections between neighboring cells interlock. Canaliculi arise between adjacent epithelia. The wall of the yolk sac thickens as a result of this infolding and the densely packed capillaries. Infoldings are especially predominant in the sac-like portion of the yolk sac, and only suggested in the ends. Involution of the yolk sac begins in the peripheral end segments and proceeds centripetally. Numerous glycogen particles appear in the yolk sac entoderm cells of the ovine fetus at a CRL of 36 mm, and by a CRL of 42 mm, the sac-like portion has also begun to show signs of degeneration. Mesenchyme is very sparse within the wall of the yolk sac throughout the entire period of development.(ABSTRACT TRUNCATED AT 400 WORDS)


[The segmental organization and horny structure of the claw of the dog]. This study classifies the segments of the dog claw and presents a new conception to define crown and wall segments that deviates from the common statements in literature. The perioplic segment extends along the internal surface of the claw fold (vallum). The crown segment lies in the unguicular groove, and there in a semicircular zone. In the crown segment there is development of horn tubuli, but these lose their tubular structure before they have reached the top of the claw. The wall segment, adjacent distally, comprises a dorsal ridge, and lateral surfaces proximally and distally which are all marked by superficial lamellar-shaped surface configurations. The sole segment shows desquamative soft horn and is situated between the free margin of the wall segment palmar/plantar to the unguicular process. The new conception of homology of dog claw distinguishes between main and secondary criteria. The main criterion is the epidermal and dermal configuration. Secondary criteria are the typical horn products (for example perioplic horn) and dimension, shape and topography of segments. The sterile/fertile-bed theory, which was believed to be the determining factor for the classification of the dog claw, is neither the proper method to define the individual segments nor to judge the rate of cornification.


[Ultrastructure of the luteal cells of female cattle]. An ultrastructural study to the corpus luteum (CL) from 263 non pregnant female cattle slaughtered in slaughter-houses of León (Spain) was done. The luteal cells are classified in five types, noted with roman numbers. We made a comparison between these cells and the cells described in the literature. We believe that luteal cells have a development from type I to V, the first type is the principal inside the developing CL. The types II and III are in the mature CL, and the two last types are in the degenerating CL. It will be the same cell in different stages of development. We did not find characteristic structures to differentiate luteal cells originating from the granulosa or the theca.


Studies on the parietal region of the cervid skull. II. The parietooccipital region in the skull of the fallow deer (Dama dama L.). In contrast to the situation in roe deer (Kierdorf and Kierdorf, in press) and other cervid species, an os interparietale was missing in the fallow deer cranium. Absence of this skull element in Dama dama is regarded as an apomorphic character state. The area covered by the interparietals in Capreolus was occupied by the parietals in Dama. This condition (loss of interparietals, enlargement of parietals) is in accord with a trend seen in vertebrate evolution, that is, progressive reduction in the number of skull elements concomitant with enlargement of the remaining bones. Synostosis of the parietals in Dama started a few days post partum and was completed at about 7 to 8 months of age. In males, obliteration of the sutura parietooccipitalis commenced in adult life, whereas in females only closure of the central region of this suture was occasionally observed.


Drug-resistant tuberculosis in an urban population including patients at risk for human immunodeficiency virus infection. In the past 5 yr, an increased incidence of tuberculosis has been noted in the United States. Simultaneously, the population infected with human immunodeficiency virus-type I (HIV-I) and the number of cases of acquired immunodeficiency syndrome (AIDS) have increased. Selected areas of the United States have also reported increases in the frequency of drug-resistant isolates of Mycobacterium tuberculosis. Because our institution serves a population in which tuberculosis, AIDS, and drug resistant isolates of M. tuberculosis are frequently encountered, we sought to better define interrelationships among these factors by retrospectively reviewing the demographic, clinical, bacteriologic, and radiologic data for all adult patients in whom M. tuberculosis was isolated from a culture of respiratory-tract secretions during a 1-year period (June 1, 1988 to May 31, 1989). Two hundred forty-six patients were thus identified; 66.5% were U.S. born blacks, and 62.6% were 17 to 40 yr of age. Risk factors for HIV infection were present in 106 patients. The overall resistance rate (one or more drugs) = 30.9%, with primary resistance = 22.6% (35 of 155) and secondary resistance = 49.2% (29 of 59). In addition, 12 resistant isolates were found in 32 patients whose prior treatment status was indeterminate. Of the resistant isolates, 56.6% (43 of 76) were multiply resistant. Isoniazid resistance was noted in 90.7% (69 of 76) and rifampin resistance was noted in 50% (38 of 76) of the resistant isolates. No significant differences in the overall frequency of resistance were noted in patients at risk for HIV infection compared with those without these risks.(ABSTRACT TRUNCATED AT 250 WORDS)


Pneumonia following closed head injury.

Pneumonia following closed head injury. Pneumonia is common among patients with artificial airways in place. Most prior studies of such pneumonia involve a heterogeneous group of patients, usually with major medical or surgical illnesses. We studied the incidence of pneumonia in a group of patients with isolated closed head injury (CHI) in an effort to determine the pattern of the problem in the absence of other injuries and to determine whether the pattern of development of pneumonia in these patients was comparable to that in more heterogeneous groups of mechanically ventilated patients. We studied 109 initially comatose patients with isolated CHI who were ventilated 24 h or more. The mean age was 30.3 +/- 20.2 yr, 72% were male, and the admission Glasgow coma score was 4.9T +/- 1.4. Overall, 45 patients (41%) developed pneumonia, with the majority (29/45) occurring during the first 3 days of hospitalization. No patient developed pneumonia after the first week despite the fact that many were still ventilated, others remained intubated, and yet others were extubated but comatose. Patients who developed pneumonia experienced a longer ICU stay (10.5 +/- 5.4 days versus 7.2 +/- 4.3 days, p = 0.001) and hospital stay (34.8 +/- 27.6 versus 22.5 +/- 20.2 days, p = 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)


A comparative study of the effects of almitrine bismesylate and lateral position during unilateral bacterial pneumonia with severe hypoxemia. The management of patients with unilateral pneumonia and severe hypoxemia often represents a therapeutic challenge. Mechanical ventilation with the diseased lung uppermost may improve gas exchange, but it is not devoid of adverse effects. No hemodynamic measurements have been reported in patients ventilated in this manner; therefore, whether or not the improvement in PaO2 is counterbalanced by hemodynamic deterioration remains unknown. Almitrine bismesylate is a drug that seems able to improve gas exchange in patients with chronic obstructive pulmonary disease or the adult respiratory distress syndrome. The increase in PaO2 after its administration has been attributed to an improvement in ventilation-perfusion relationships. Its use has never been reported during unilateral pneumonia with severe hypoxemia. We therefore compared its effects with those of lateral position in eight consecutive mechanically ventilated patients with unilateral pneumonia. Blood gas and hemodynamic measurements were performed both at maintenance FIO2 and at an FIO2 of 1.0. Almitrine (1 mg/kg over 1 h) had no effect on PaO2 under either FIO2 condition. Cardiac output remained unchanged, but mean pulmonary artery pressure increased from 22.5 +/- 1.2 to 26.5 +/- 1.3 mm Hg (p < 0.02). By contrast, lateral position had striking effects on PaO2, which increased from 100 +/- 14 mm Hg in supine position to 156 +/- 23 mm Hg (p < 0.01) when the abnormal lung was placed uppermost at maintenance FIO2 and from 207 +/- 21 (supine) to 300 +/- 28 mm Hg (lateral) (p < 0.01) at FIO2 1.0.(ABSTRACT TRUNCATED AT 250 WORDS)


Prone position alters the effect of volume overload on regional pleural pressures and improves hypoxemia in pigs in vivo. Oxygenation improves in patients with adult respiratory distress syndrome and in animals with oleic acid-induced lung injury when they are turned from the supine to the prone position. Dependent and nondependent pleural pressures (Ppl) were measured in six pigs ventilated in the supine and prone positions before and after volume infusion (VI). Before VI the mean +/- SEM AaPO2 difference was 26 +/- 8 mm Hg when the animals were supine and 10 +/- 2 mm Hg when they were prone (p > 0.05). After VI the AaPO2 was 64 +/- 6 mm Hg when the animals were supine (p < 0.05) and 43 +/- 7 mm Hg when they were prone (p < 0.05). VI increased the Ppl gradient from 0.53 +/- 0.1 to 0.71 +/- 0.1 cm H2O/cm when the animals were supine (p < 0.05) and from 0.17 +/- 0.1 to 0.27 +/- 0.1 cm H2O/cm when they were prone (p < 0.05). Dependent Ppl at FRC was much less positive when the animals were prone versus supine (0.9 +/- 0.3 versus 3.0 +/- 0.5 cm H2O, p < 0.05), suggesting that the airways in these dependent regions would narrow and/or close and that ventilation to these regions would diminish as a result of VI.


Pulmonary vascular pressure-flow characteristics. Effects of dopamine before and after pulmonary embolism. We compared the general hemodynamic effects of dopamine and dobutamine in dogs with acute pulmonary hypertension complicated by a decrease in cardiac output (CO). The pulmonary hypertension was induced by injection of autologous blood clot. Emboli markedly increased mean pulmonary artery pressure (Ppa) and decreased CO (both p < 0.001). Both dopamine and dobutamine increased CO 50% (p < 0.05) and decreased pulmonary vascular resistance (PVR) (p < 0.05), calculated as (PAP - left ventricular end diastolic pressure)/CO. Mean PVR (mm Hg/L/min) decreased from 16.1 to 12.4 with dopamine and from 16. to 11.9 with dobutamine, both p < 0.05. Ventricular filling pressures were not affected. In another 12 dogs we investigated the effects of both drugs on pulmonary pressure-flow (P-Q) characteristics. P-Q characteristics were determined in dogs with normal Ppa values and in those with embolic pulmonary hypertension. The slope of the P-Q relationship defines the incremental vascular resistance and the extrapolated pressure intercept, the effective vascular outflow pressure. All P-Q relationships were described well by a linear equation. Despite significant systemic effects in both groups and despite a decrease in PVR with both drugs in embolized dogs, neither drug significantly affected pulmonary P-Q characteristics. The discrepancy between PVR and incremental resistance is explained by an incorrect assumption in PVR that the left ventricular filling pressure is the effective vascular outflow pressure. We conclude that both before and after the induction of pulmonary hypertension, both dopamine and dobutamine improve CO without affecting pulmonary vascular tone.


Interaction between effects of hypoxia and hypercapnia on altering left ventricular relaxation and chamber stiffness in dogs. Left ventricular (LV) isovolumic relaxation and chamber stiffness were examined in dogs to see how hypoxemia and hypercapnia occurring during an acute exacerbation of chronic obstructive lung disease could directly affect diastolic performance. Measurements were obtained during baseline (arterial PO2 approximately 200 mm Hg; PCO2 approximately 35 mm Hg; pH approximately 7.35), hypoxia (PO2 approximately 35 mm Hg; PCO2 approximately 35 mm Hg; pH approximately 7.35), and hypoxia-hypercapnia (PO2 approximately 35 mm Hg; PCO2 approximately 60 mm Hg; pH approximately 7.15) in an open-chest, open-pericardium preparation. Changes in LV diastolic performance during hypoxic and hypercapnic interventions were contrasted with those caused by ventricular interdependence in which pulmonary artery occlusion was used to produce a right to left ventricular septal shift. Subendocardial ultrasonic crystal transducers were placed along the three orthogonal axes of the left ventricle to measure dimensions by sonomicrometry. Along each axis, LV end-diastolic dimension-pressure relations were constructed in the various conditions; the slope of this relationship, obtained over a linear portion of the curve, was used as an index of diastolic dimensional distensibility (DD). The results showed that during hypoxia, DD appeared decreased along the three axes, with a reduction in filling primarily along the anterior-posterior and apex-base axes. The addition of hypercapnia to hypoxia completely abolished the hypoxic effect. On the other hand, during pulmonary artery occlusion, LV DD were not changed along the latter two dimensions. We conclude that during respiratory failure, LV diastolic performance may be directly affected by arterial blood gas tensions, which by altering the interaction between the contractile filaments modulate LV filling.


Cardiovascular effects of periodic occlusions of the upper airways in dogs. Hypoxemia and decreased intrathoracic pressure have been postulated as contributing causes of cardiovascular morbidity in obstructive sleep apnea syndrome (OSAS). Because of the difficulty of manipulating experimental conditions in humans, we developed an anesthetized closed-chest dog model, simulating the periodic airway occlusions of OSAS by periodic occlusions of the endotracheal tube (PUO). Using a periodicity of 60 s occluded, followed by 60 s ventilation for five to seven cycles, we measured heart rate (HR), cardiac output (CO), arterial pressure (Pa); left ventricular (LV) end-diastolic and end-systolic transmural pressure; dp/dt of LV pressure; left anterior descending (LAD) coronary blood flow (CBF), and regional myocardial contractility and intramyocardial pH. Four experimental conditions were studied: room air (RA) breathing (PO2 = 40); 100% O2 breathing (O2), and RA and O2 breathing with critical LAD stenosis (CS). Under all conditions PUO produced decreases in CO (10 to 30%) and proportional decreases in Pa. HR decreased, and in all but RA conditions stroke volume was unchanged. During the obstructed phase, indices of LV preload decreased. Indices of LV afterload also decreased except for LAD-perfused myocardium under RACS conditions. This latter was shown to be associated with regional ischemia (decreased regional pH and shortening). Regional ischemia was also demonstrated in two of nine dogs even under O2CS conditions. Among our major conclusions: (1) decreased Pa during PUO is due to decreased CO; (2) LV afterload does not increase during PUO; (3) with limited coronary flow reserve (CS), PUO can lead to myocardial ischemia. This is mostly but not solely due to hypoxia.


Effect of hyperoxia on the arousal response to airway occlusion during sleep in normal subjects. The effect of hyperoxia on the arousal response to airway occlusion during non-rapid eye movement (NREM) sleep was studied in six normal male subjects with a mean age (+/- SD) of 23.5 +/- 8.7 yr by testing the response to the occlusion of a face mask covering the nose and mouth. Occlusion trials while the subjects breathed room air (room air condition) were alternated with trials in which subjects breathed a mixture of room air and oxygen adjusted to maintain a sleeping baseline arterial oxygen saturation of 98% (hyperoxic condition). The time to arousal (mean +/- SEM) was significantly longer during oxygen administration (4.1 +/- 4.5 versus 28.9 +/- 4.6 s; p < 0.002). The maximal deflections in airway pressure were measured at a supraglottic location during airway occlusion to reflect the degree of inspiratory effort. The maximal airway suction pressure preceding arousal did not differ between the room air (27.4 +/- 5.4 cm H2O) and hyperoxic conditions (26.6 +/- 5.9 cm H2O). Conversely, the rate of increase in inspiratory effort (maximal pressure) during occlusion was decreased by oxygen administration. We conclude that hyperoxia prolongs the time to arousal after airway occlusion by decreasing the rate of increase in the magnitude of inspiratory efforts, but it does not change the arousal threshold.


Fast-CT evaluation of the effect of lung volume on upper airway size and function in normal men. The present study was performed to evaluate the regional changes in pharyngeal cross-sectional area (CSA) that occur with changes in lung volume in normal men. Fast-CT and genioglossal electromyogram (EMGgg) were used to study upper airway (UA) size and dilator muscle activity at TLC, lung volume at end-tidal inspiration (VTei), FRC, and residual volume (RV) in 30 men with a mean age of 46 +/- 3 yr and no significant sleep-disordered breathing, mean AHI = 4 +/- 1 per hour. Compared with values at FRC, minimum CSA (Amin) increased 154 +/- 31% at TLC (p = 0.0001), 19 +/- 10% at VTei (p = 0.03), and there was a trend toward a decrease of 31 +/- 12% at RV (p = 0.07). Similar but smaller changes were observed in mean CSA (Amean), with an increase of 69 +/- 14% at TLC (p = 0.0001), 8 +/- 5% at VTei (p = 0.01), and a decrease of 17 +/- 7% at RV (p = 0.01). Both the velopharyngeal (VP) and orohypopharyngeal (OHP) segments of the UA increased in size with increasing lung volume. Both Amin and Amean of the OHP segment at TLC were larger (55 +/- 19 and 38 +/- 14%, respectively) than the respective measurements in the VP segment. EMGgg activity doubled from 12 +/- 1 microV at FRC to 25 +/- 1 microV at TLC (p = 0.006). There was no change in EMGgg with tidal ventilation or with exhalation to RV. Changes in CSA directly paralleled changes in lung volume in this group of normal awake nonobese men.(ABSTRACT TRUNCATED AT 250 WORDS)


Local application of atropine attenuates the upper airway reaction to cold, dry air. In some individuals, inhalation of cold, dry air (CDA) provokes symptoms of rhinitis, accompanied by an increase in the levels of inflammatory mediators and markers of plasma leakage of recovered nasal lavages. Because rhinorrhea is a major component of this reaction and because nasal glands are heavily innervated by the parasympathetic system, we assessed the effect of atropine on the nasal reaction to CDA. Using a double-blind, randomized, crossover design, we administered a total dose of 0.5 mg of atropine or placebo intranasally to 18 volunteers before provocation with CDA. The reaction was monitored with symptom scores and by measuring the concentrations of histamine, N-alpha-p-tosyl-L-arginine methyl ester (TAME)-esterase activity and albumin, as well as the osmolality of lavage fluids before and after the provocation. Atropine significantly reduced rhinorrhea, the levels of histamine, and TAME-esterase activity as well as the osmolality of recovered lavage fluids, but had no effect on nasal congestion or albumin. Even with atropine, however, rhinorrhea and TAME-esterase activity were still significantly increased over the prechallenge baseline. Our results demonstrate that atropine-sensitive parasympathetic efferent pathways contribute to the CDA-induced rhinitis. We speculate that (1) the glandular and the vascular events of the upper airway reaction to dry air have different pathophysiologic mechanisms; (2) a significant component of TAME-esterase activity in lavage fluids may be of glandular origin; and (3) in addition to parasympathetic nerve activation, other mechanisms are involved in the upper airway reaction to dry air. The mechanism(s) leading to the reduction of histamine is unknown.


Capsaicin-induced cough in humans.

Capsaicin-induced cough in humans. We have evaluated the properties of capsaicin as a selective cough-inducing agent in healthy human subjects. Despite frequent coughing, the subjects could inhale repeated breaths of capsaicin aerosol during 60 s without difficulty. Cough started immediately on inhalation and was most intense during the first 30 s. Cough always disappeared promptly when the capsaicin inhalation was terminated. The cough response was well reproducible and concentration-dependent up to 10 microM; at higher concentrations there was a distinct plateau of the cough response. Specific airway conductance was not changed 3 min after 50 microM capsaicin. Capsaicin (> or = 10 microM) had a burning taste, but there were no visual signs of pharyngitis or laryngitis. Citric acid (nebulized solutions 0.125 to 32%) had a choking effect and could be administered only as single breaths. There was no correlation between the cough response to citric acid and to capsaicin. Inhaled lidocaine (20 and 80 mg from nebulized solutions) caused a dose-dependent inhibition of capsaicin-induced cough. Lidocaine suppressed citric acid-induced cough as effectively as capsaicin-induced cough. In conclusion, we have characterized capsaicin-induced cough and demonstrated that it can be a useful tool in the study of cough reactivity and for evaluation of antitussive agents in humans. Capsaicin may be complementary to citric acid and may offer experimental advantages over this traditional tussive stimulus.


Inflammatory response to acute inhalation of endotoxin in asthmatic patients. Inhalation of 20 micrograms endotoxins (from the membrane of Gram-negative bacteria) has been reported to induce a bronchial obstructive response in asthmatic subjects. The aim of the present study was to evaluate in asthmatic patients the possibility of an inflammatory response to inhaled endotoxins. Eight patients with mild asthma were submitted to bronchial challenge tests, in a single-blind trial, on Day 1 with control solution and on Day 7 with 20 micrograms endotoxin of Escherichia coli (026:B6). Local inflammatory response was indirectly evaluated by the degree of bronchial hyperresponsiveness (BHR) expressed as PD20 FEV1 histamine (the dose of histamine inducing a 20% decrease in FEV1) at 0, 6, 24, and 48 h and 7 days. Systemic inflammation was investigated by sequential blood determinations of total (and differential) white cells, complement anaphylatoxin C5a, interleukin-6 (IL-6), tumor necrosis factor alpha (TNF-alpha), and C-reactive protein (CRP). A significant (p < 0.01) bronchial obstructive response was demonstrable 45 min after lipopolysaccharide (LPS) inhalation, lasting 5 h. Comparing the level of BHR after control inhalation, a significant (p < 0.05) increase in BHR was shown 6 h after LPS, partially normalized at 24 and 48 h. A short peak in TNF-alpha at 60 min (p < 0.05) and an increase in total white blood cells (p < 0.01) and neutrophil polymorphonuclear neutrophils at 360 min (p < 0.05) and of CRP at 24 and 48 h (p < 0.05 and p < 0.01) were significant. The other blood parameters did not change significantly.(ABSTRACT TRUNCATED AT 250 WORDS)


Bronchodilation with a potent and selective leukotriene D4 (LTD4) receptor antagonist (MK-571) in patients with asthma. The sulfidopeptide leukotrienes LTC4, LTD4, and LTE4 can cause airway smooth muscle contraction and have been implicated in the pathophysiology of asthma. MK-571 is a selective, potent LTD4 receptor antagonist that could attenuate airway obstruction in asthma by inhibiting the actions of sulfidopeptides at the LTD4 receptor site. The objectives of this study were to investigate the potential for MK-571 to cause bronchodilation in asthma patients with existing airway obstruction and to evaluate its effect on the bronchodilation response to an inhaled beta 2-agonist (albuterol). Twelve male patients (ages 19 to 42 yr) with asthma (baseline FEV1 50 to 80% predicted) participated in this placebo-controlled, randomized, two-period, cross-over study. On separate treatment days, each patient received either MK-571 or placebo intravenously for 6 h; inhaled albuterol was administered at the fifth and sixth hour of MK-571/placebo treatment to achieve maximal bronchodilation on that study day. Spirometry (forced expiratory volume in 1 s, FEV1) was monitored at intervals throughout each study period. MK-571 caused clinically significant bronchodilation; the increase in FEV1 above baseline, 20 min after the start of the MK-571 infusion, was 22 +/- 3.9% compared with 1.3 +/- 2.3% for placebo (mean +/- SE, p < 0.01). This degree of bronchodilation was maintained throughout the MK-571 infusion. In addition, bronchodilation from inhaled albuterol appeared additive with MK-571. Finally, baseline airway obstruction correlated with the degree of bronchodilation achieved with MK-571 (r = -0.73; p = 0.007).(ABSTRACT TRUNCATED AT 250 WORDS)


Effect of inhaled furosemide and torasemide on bronchial response to ultrasonically nebulized distilled water in asthmatic subjects. Inhaled furosemide has been shown to reduce the bronchoconstriction induced by several indirect stimuli, including ultrasonically nebulized distilled water (UNDW). Because the protective effect could be due to the inhibition of the Na(+)-2Cl(-)-K+ cotransport system of bronchial epithelium, we have compared the protective effect of inhaled furosemide with that of inhaled torasemide, a new and more potent loop diuretic, on UNDW-induced bronchoconstriction in a group of 12 asthmatic subjects. UNDW challenge was performed by constructing a stimulus-response curve with five increasing volume outputs of distilled water (from 0.5 to 5.2 ml/min) and the bronchial response expressed as the provocative output causing a 20% fall in FEV1 (PO20UNDW). On different days, each subject inhaled an equal dose (28 mg) of furosemide and torasemide in a randomized, double-blind, placebo-controlled study 5 min prior to an UNDW challenge. Furosemide and torasemide had no significant effect on resting lung function. The geometric mean value of PO20UNDW measured after placebo was 1.73 ml/min. This was significantly lower than that recorded after furosemide (4.25 ml/min; p < 0.025), but not after torasemide (3.05 ml/min; p = 0.07). Inhaled furosemide totally blocked bronchial response to UNDW in five subjects. In two of five subjects the response was also blocked by inhaled torasemide. A remarkable increase in diuresis was noted only after torasemide in most subjects. We conclude that inhaled furosemide has a better protective effect than does inhaled torasemide against UNDW-induced bronchoconstriction. However, the protective effect of furosemide is variable, with some asthmatic patients showing no change in bronchial response to UNDW.(ABSTRACT TRUNCATED AT 250 WORDS)


Antigen-induced acute and late-phase responses in primates. We have examined the proinflammatory cell influx as well as the levels of eosinophil and neutrophil-derived granule proteins in BAL fluid obtained from monkeys undergoing acute and late-phase (dual) or single acute bronchoconstriction following antigen inhalation. Prior to antigen inhalation, there was a significantly higher number (and percentage) of eosinophils in BAL fluid from dual responder monkeys as compared with single responders. The late-phase response (LPR) (6 to 8 h postantigen) was associated with a decrease in the number of BAL eosinophils and an increase in the levels of BAL fluid EPO that returned to baseline levels by 24 h postantigen inhalation. In contrast, the number of BAL neutrophils prior to antigen inhalation were low. Concurrent with the LPR, the number of BAL neutrophils and the concentration of EPO in BAL fluid were significantly increased above that occurring in single responders. Chronic treatment (7 days) with dexamethasone significantly reduced the number of BAL eosinophils and the BAL levels of EPO prior to antigen inhalation in dual responder (LPR) monkeys and significantly blocked the dual response and both the associated neutrophil influx into the airways and an increase in BAL fluid EPO during the LPR. We conclude that, in this primate model, eosinophil activation and a large influx of neutrophils into the airways is associated with the occurrence of the antigen-induced late-phase airway obstructive response.


Delivery of a nebulized aerosol to a lung model during mechanical ventilation. Effect of ventilator settings and nebulizer type, position, and volume of fill. Several factors may affect the delivery of a nebulized aerosol to the lung through an endotracheal tube during mechanical ventilation. To study these factors in vitro, a model representing ventilation of an adult patient was constructed by linking a Servo 900C ventilator to a standard humidified circuit and an endotracheal (ET) tube positioned within a pipe representing the trachea. This was connected via a filter to a lung simulator. Nebulizers filled with 99mTc human serum albumin were positioned in the circuit, and the delivery of nebulized aerosol through the ET tube into the filter was measured using a gamma camera. With the use of an inspiratory phase-activated System 22 Acorn jet nebulizer, typical adult ventilator settings, and a 3-ml nebulizer solution volume, 5.4% of the nebulizer dose reached beyond the end of the ET tube. This was increased by increasing the inspiratory time, reducing the respiratory rate or respiratory minute volume, and by repositioning the nebulizer on the inspiratory limb of the Y-piece and was reduced by slowing the driving gas flow to the nebulizer. Under the same conditions, delivery was 3.1 and 4.4% using the Samsonic and Fisoneb ultrasonic nebulizers, respectively. Increasing the fill volume and the addition of an aerosol storage chamber increased delivery with all three nebulizers. These experiments suggest some simple ways of improving aerosol delivery during mechanical ventilation, including increasing the volume of nebulizer fill, repositioning the nebulizer in the ventilator circuit, adding an aerosol storage chamber, and adjusting ventilator settings to maximize delivery.


Aerosolized beclomethasone in chronic bronchitis. Improved pulmonary function and diminished airway inflammation. Chronic bronchitis is associated with airways obstruction and inflammation. In order to determine whether aerosolized beclomethasone can modulate airway inflammation and diminish airway obstruction, subjects with chronic bronchitis performed spirometry and underwent bronchoalveolar lavage (BAL) before and after receiving 6 wk of therapy (five puffs four times a day) with either aerosolized beclomethasone (n = 20) or placebo (n = 10) in a double-blinded, randomized fashion. All subjects received aerosolized albuterol before each use of the study medications. Before BAL, the airways were visually assessed for the appearance of inflammation and assigned a score, the bronchitis index. BAL was performed by instilling five 20-ml aliquots of saline into each of three sites and pooling and separately analyzing the returns from the first aliquots to yield a "bronchial sample." The bronchial lavages were repeated in an additional three sites to increase the volume of fluid available for analysis. The fluid was prepared for cytologic examination by cytocentrifugation. Albumin (as a measure of epithelium permeability) and lactoferrin and lysozyme (as measures of serous cell activity) were measured in unconcentrated BAL fluid by enzyme-linked immunosorbent assay, and concentrations in epithelial lining fluid were estimated using urea as an internal marker for dilution. After treatment, the beclomethasone group, but not the placebo group, showed improvement in FVC (p = 0.02), FEV1 (p = 0.002), and 25 to 75% forced expiratory flow (p = 0.006). Associated with the improvement in spirometry, the bronchitis index fell (13.5 +/- 1.0 versus 10.75 +/- 1.1, p = 0.02) in the beclomethasone-treated group, but not the placebo-treated group.(ABSTRACT TRUNCATED AT 250 WORDS)


Perception of airway obstruction in a random population sample. Relationship to airway hyperresponsiveness in the absence of respiratory symptoms. Subjects with asymptomatic airway hyperresponsiveness in epidemiologic studies may have variable airway obstruction that is not perceived as dyspnea. We tested the hypothesis that such subjects are less likely to report an increase in dyspnea during histamine-induced bronchoconstriction than symptomatic hyperresponders. A random population sample of 412 middle-aged subjects was studied. Before and after a standardized histamine challenge test, subjects recorded Borg scores for dyspnea: any increase in Borg score was considered significant. More than 80% of hyperresponsive subjects (PC10 histamine < or = 16 mg/ml) had no symptoms. The presence of prechallenge dyspnea was related to increased airways responsiveness and current smoking. An increase in Borg score was associated with younger age, more severe airway responsiveness, atopy, and female sex. The level of and increase in the Borg score were not significantly related to level and change in airway caliber (FEV1). In hyperresponsive subjects (PC10 < or = 16 mg/ml), subjects who reported dyspnea, wheeze, or asthma were more likely to show an increase in Borg score during histamine provocation than asymptomatic subjects (adjusted odds ratio 4.01, 95% confidence interval 1.01 to 16.00, p = 0.049), after adjustment for age, sex, smoking habits, FEV1, and atopy. This suggests that asymptomatic hyperresponders may have variable airway obstruction that is not recognized as breathlessness.


The interaction between immunoglobulin E and smoking in airflow obstruction in the elderly. Airflow obstruction and serum immunoglobulin E (IgE) were assessed in 250 men and women 65 to 91 yr of age who had been selected from a larger general population sample according to reported respiratory symptoms. After allowance for age and sex, serum IgE and smoking interacted synergistically as risk factors for airflow obstruction such that on average an IgE > or = 81 IU/ml in current smokers was associated with a FEV1/FVC ratio 14.4 percentage points (95% CI, 8.8 to 19.9) less than in lifelong nonsmokers with an IgE < or = 10 IU/ml. This synergistic interaction was apparent in subjects who showed no evidence of airway lability (i.e., no hyperresponsiveness to inhaled methacholine or, if bronchial hyperresponsiveness could not be tested, no clear improvement in airflow obstruction after inhalation of salbutamol). The findings suggest that the role of IgE in the pathogenesis of airflow obstruction is not confined to asthmatics.


Changes in interleukin-1 and tumor necrosis factor production by peripheral blood monocytes after specific bronchoprovocation test in occupational asthma. The pathogenetic mechanisms of occupational asthma (OA) due to low-molecular-weight compounds have been poorly defined, and further studies are required to clarify the role of immunologic mechanisms in OA. Until now cellular mechanisms have been less investigated than humoral ones. We have evaluated interleukin-1 (IL-1) and tumor necrosis factor (TNF) production by peripheral blood monocytes (PBM), and peripheral T-cell subpopulations in 22 subjects with possible OA before and after specific bronchoprovocation test (SBPT). After SBPT, three subjects had an immediate reaction, seven a late reaction, and two a dual reaction. Ten subjects had no asthmatic reaction to SBPT. Spontaneous release of IL-1 from PBM did not change significantly after SBPT. TNF activity was increased 48 h after SBPT in immediate reactions and 72 h after SBPT in late-dual reactions. These results suggest that exposure to occupational agents may induce activation of PBM with increased spontaneous release of cytokines, such as TNF.


Exposure-response relationships between occupational exposures and chronic respiratory illness: a community-based study. Data from a random sample of 3,606 adults 40 to 69 yr of age residing in Beijing, China, were analyzed to investigate the association of reported occupational exposures to dusts and gases/fumes with the prevalence of chronic respiratory symptoms and level of pulmonary function. The prevalence of occupational dust exposure was 32%, and gas or fume exposure, 19%. After we adjusted for age, sex, area of residence, smoking status, coal stove heating, and education, an increased prevalence of chronic phlegm and breathlessness was significantly related to both types of exposures. Chronic cough was significantly related only to dust exposure, and persistent wheeze only to fume exposure. The global estimates of the relative odds of the four symptoms were 1.30 (95% CI [confidence interval] 1.09 to 1.48) and 1.27 (95% CI 1.09 to 1.48), respectively, for dusts and for gases/fumes. These two occupational exposures are associated with chronic respiratory symptoms independent of smoking, gender, and each other. There was an increasing prevalence of each symptom with increasing dust and fume exposure, represented by the index of cumulative exposure duration and exposure intensity. Linear trends for increased prevalence of chronic bronchitis and breathlessness were significant for both exposures, while the linear trend for wheeze was only significant for gases/fumes. Among subjects who did not report using coal stove heating, dust exposure was a significant predictor for FEV1, FEV1/FVC, FEF25-75, and peak expiratory flow rate (PEFR). There was also a significant decrease for FEV1 and FVC with increase of gas/fume exposure levels. Both current and former smokers appeared to be more susceptible to the effect of dusts than the never smokers.(ABSTRACT TRUNCATED AT 250 WORDS)


A multicenter registry of patients with acute respiratory distress syndrome. Physiology and outcome. In a multicenter registry conducted over 2 yr of patients with acute respiratory distress syndrome (ARDS), we enrolled 153 patients and collected data daily for 7 consecutive days and weekly thereafter until death or hospital discharge. The purposes of the registry were (1) to determine whether a more liberal definition of ARDS (PaO2/FIO2 < or = 250; bilateral pulmonary infiltrates within 7 days) than those commonly used would result in enrollment of patients earlier in their clinical course, and (2) to study the clinical course of the syndrome in survivors and nonsurvivors. The mortality rate was 54% and it was significantly greater in older versus younger patients (75% versus 37%) and in septic versus nonseptic patients (60% versus 43%). We found that the definition of ARDS used for the registry resulted in enrollment of patients 1 to 7 days earlier than was the case when other published definitions of ARDS were applied to the patient population. Fewer than 2% of the patients failed to meet one of the nonregistry definitions of ARDS within 7 days. The mortality rate was independent of the definition used to identify ARDS patients. Our results suggest that a more liberal definition of ARDS than those commonly used can result in identification of the same population of patients earlier in their clinical course.


Elevated levels of NAP-1/interleukin-8 are present in the airspaces of patients with the adult respiratory distress syndrome and are associated with increased mortality. The adult respiratory distress syndrome (ARDS) is characterized by increased neutrophils within the airspaces of the lungs. In order to determine if neutrophil activating protein (NAP)-1/interleukin-8 (NAP-1/IL-8) could be an important cause of neutrophil influx and activation in ARDS, we examined fluid, which was either directly aspirated or lavaged with saline from the lungs of patients with ARDS. NAP-1/IL-8 was present in significantly higher concentrations in the fluids of patients with ARDS compared with control subjects. There was a significant correlation between the percentage of neutrophils in the lavage fluids and the NAP-1/IL-8 concentration (r2 = 0.74). Furthermore, the NAP-1/IL-8 concentration of the pulmonary edema fluid was equivalent to the optimal concentration required to induce neutrophil chemotaxis in vitro. Although not all of the chemotactic activity of the edema fluid was removed by an anti-NAP-1/IL-8 affinity column, the data established that NAP-1/IL-8 is an important neutrophil chemotaxin in the airspaces of patients with ARDS. In addition, those patients with very high concentrations of NAP-1/IL-8 in their bronchoalveolar lavage fluids had a higher mortality rate than those patients with lower concentrations of NAP-1/IL-8. The correlation between NAP-1/IL-8 concentration and mortality is not paralleled by total protein concentration and mortality.


Airway inflammation predicts diffuse alveolar hemorrhage during bone marrow transplantation in patients with Hodgkin disease. We determined risk factors present in patients with Hodgkin disease that predicted the development of diffuse alveolar hemorrhage syndrome (DAH) during autologous bone marrow transplantation (BMT). One hundred twenty-three patients with Hodgkin disease prospectively underwent bronchoscopy with bronchoalveolar lavage (BAL) before receiving BMT. The bronchitis index (BI) of the airways and bronchial and alveolar cell counts and differentials were determined in all patients and compared with 20 normal nonsmoking volunteers. Logistic regression analysis was used to determine factors that predicted for the development of DAH. Visual evidence of bronchial injury was observed regardless of smoking history (BI = 7.8 +/- 0.5 for BMT versus 2.3 +/- 0.5 for volunteers, p = 0.001). BMT patients who developed DAH (n = 14) had significantly greater numbers of bronchial neutrophils and eosinophils compared with DAH-negative (n = 109) patients (bronchial polymorphonuclear leukocytes (PMN), 33 +/- 7% versus 14 +/- 2%, p = 0.006; bronchial eosinophils, 0.9 +/- 0.3% versus 0.4% +/- 0.07%, p = 0.02). Logistic regression analysis revealed that the presence of bronchial PMN > 20% or bronchial eosinophils > zero% were predictive of DAH (p = 0.005 and 0.05, respectively). When both predictors were positive, the rate of DAH was 10 times greater than when both predictors were negative (43% versus 4% DAH occurrence). Survival was also significantly reduced when these predictors were positive. This study demonstrates that bronchial inflammation is present with or without intraluminal inflammatory cells in the majority of patients with Hodgkin disease before BMT. The subgroup of these patients with increased bronchial inflammatory cells are at greatly increased risk for development of DAH and death.


The origin of water and urea sampled at bronchoalveolar lavage in asthmatic and control subjects. Bronchoalveolar lavage (BAL) urea has been advocated as a denominator that might allow for the dilution of the pulmonary epithelial lining fluid sampled at BAL, and so provide a meaningful method of expressing BAL data. We investigated the origin of water and urea sampled at BAL in five asthmatic and five control subjects using radiolabeled urea injected intravenously 5 min before BAL. Labeled BAL urea was found to be fully equilibrated with that in the bloodstream. A strong relationship was found between influx of radiolabeled water and radiolabeled urea from blood to BAL fluid, suggesting that urea sampled at BAL may be derived predominantly from an acute movement from the bloodstream into the BAL aspirate. We conclude that urea is an inappropriate denominator for the expression of BAL results, and that the fluid and solute dynamics that occur during BAL are both complex and variable.


Bronchoalveolar lavage cellularity. The distribution in normal volunteers. Bronchoalveolar lavage (BAL) is used to obtain inflammatory cells from the lung. For clinical research, parametric statistics are frequently used to compare cells present in BAL of patients with lung disease with cells present in BAL of normal subjects. To determine if these populations can be compared in this manner we performed BAL on 111 never-smoking, normal volunteers and determined: (1) the mean, median, standard deviation, and range of the cells in BAL; (2) whether the data are normally distributed and satisfy the criteria for use of parametric statistical analysis. The BAL cellularity was expressed as a percentage of total cells, cells per milliliter return, and total cells per lavage. Regardless of the means of expression, no measure of BAL cellularity (total cells, macrophages, lymphocytes, neutrophils, or eosinophils) conformed to the normal (bell-shaped) distribution when tested for goodness of fit with the G statistic (all p < 0.001). The lack of fit to the normal distribution was not substantially altered by either the method of expressing the data (i.e., cells per milliliter, total cells, or percent of cells) or log transformation of the data. The poor fit in all cases resulted from clumping of the data about the mean and large tails. The percent of cells were, therefore, tested for goodness of fit to the Poisson distribution, a distribution of discrete variables. The neutrophil and eosinophil percentages resulted in an excellent fit to the Poisson distribution, but macrophage and lymphocyte percentages did not.(ABSTRACT TRUNCATED AT 250 WORDS)


Distribution of lung cell numbers and volumes between alveolar and nonalveolar tissue. Although total cell number has been determined for the alveolar region of the lungs of many species, it has not been calculated for the nonalveolar lung tissues. The oriented structure of airways and vessels makes the numerical assessment of cells in nonalveolar tissues difficult. This has led many investigators to use the number of cells in the alveolar region as a direct estimate of total lung cell number. To determine the number of cells in the nonalveolar lung tissues, the lungs of eight rats weighing 230 to 380 g were inflation-fixed and embedded in araldite, and 1.5-microns serial sections of the entire left lobe were cut and stained with methylene blue for light microscopy. The sections were then uniformly point-counted using computer-controlled distances between the fields to determine the fraction of points falling on air, blood, cellular tissue, and noncellular tissue for both the alveolar and the nonalveolar regions. The total volume of cell nuclei in each compartment was determined, and the total number of cells was calculated by dividing the total nuclear volume by the mean cell nuclear volume. It was found that 87% of the lung volume was alveolar, of which 6% was tissue and contained 725 x 10(6) cells. The nonalveolar region constituted 13% of the lung volume, of which 23% was tissue and contained 250 x 10(6) cells. The average rat lung therefore contains 975,000,000 cells, of which 74% was in alveolar tissues and 26% in nonalveolar tissues. On the basis of assays of isolated lung cells, there is an average of 7 pg DNA/cell.(ABSTRACT TRUNCATED AT 250 WORDS)


Covalently linking a peptidyl carbamate elastase inhibitor to a hydrophilic polymer increases its effectiveness in preventing emphysema and secretory cell metaplasia in the hamster. A peptidyl carbamate, p-nitrophenyl N-(succinyl-L-alanyl-L-alanyl-L-prolyl-methyl)-N-isopropylcarbamate++ + (PCI) was tested for its ability to inhibit the elastolytic activity of human neutrophil elastase (HNE) and to prevent HNE-induced emphysema and secretory cell metaplasia in the hamster. In vitro, 50% of the elastolytic activity of 10 micrograms of HNE was inhibited by 0.9 micrograms of PCI, a molar ratio of PCI to HNE of 4.5. Bronchoalveolar lavage of hamsters receiving PCI intratracheally showed a rapid decrease in HNE inhibitory activity (4 min for 50% decrease), suggesting rapid clearance, binding, or inactivation of the PCI. Instillation of 300 micrograms of HNE combined with 100, 500, or 3,000 micrograms PCI, a 16-, 83-, and 503-fold molar excess of PCI, respectively (molar ratios of 17, 84, and 504), suppressed HNE-induced lung hemorrhage, but it did not moderate HNE-induced emphysema despite the large molar excess of inhibitor. When PCI was covalently bound to a linear hydrophilic polymer of alpha,beta-poly[N(2-hydroxyethyl)-D,L-aspartamide], producing a polymer-bound carbamate inhibitor (PPCI) of HNE, the time for a 50% decrease of PPCI functional activity from the hamster lung lavage was 421 min. Instillation of 100 micrograms of PPCI 1 h before instillation of 300 micrograms HNE resulted in significant amelioration of emphysema; 900 micrograms of PPCI was required to obtain amelioration of bronchial secretory cell metaplasia. The larger dose of PPCI also provided significant amelioration of emphysema when the interval between PPCI and HNE administration was 8 h.(ABSTRACT TRUNCATED AT 250 WORDS)


Monocyte inflammatory mediators impair in vitro hamster diaphragm contractility. Sepsis has been shown to impair ventilatory muscle function. To determine whether this can be attributed to direct effects of inflammatory mediators on muscle fibers, we carried out in vitro studies on hamster costal diaphragm. Baseline measurements included supramaximal peak twitch (Pt) and tetanic tension (Po), twitch half relaxation time (1/2RT) and time-to-peak tension (TTP), and force frequency response (15 to 80 Hz). Fatigability was evaluated using 60-Hz stimulations at a duty cycle of 0.4 until tension fell to 50% of baseline. Preparations were then incubated in one of the following for 60 min: (1) Krebs solution (n = 5), (2) nonstimulated monocyte supernatant (n = 5), or (3) lipopolysaccharide-stimulated monocyte supernatant (n = 5). Baseline Pt, Po, 1/2RT, TTP, force frequency response, and fatigue profile were similar between groups. After incubation there was a significant fall in Pt (mean +/- SD, 538 +/- 65 to 288 +/- 13 g/cm2, p < 0.05) and Po (1,268 +/- 132 to 921 +/- 64 g/cm2, p < 0.05) in the LPS group, with no change in the other groups. There was no change in TTP; however, 1/2RT was lower in the LPS-stimulated group after incubation (p < 0.05). There was a rightward shift in the force frequency response for the LPS-stimulated group (p < 0.05). When normalizing for initial Po, there was no significant change in the time to fatigue for any of the three groups. This study demonstrated that monocyte secretory products impair diaphragmatic contractility in vitro by a direct effect on muscle fibers.(ABSTRACT TRUNCATED AT 250 WORDS)


Inspiratory muscle weakness and dyspnea in chronic heart failure. Dyspnea is a common, disabling symptom in chronic heart failure, yet the underlying mechanisms remain unknown. The respiratory muscle pump is composed of skeletal muscles whose strength directly influences the pump's performance. Respiratory muscle weakness is important in the dyspnea experienced by some patients with pulmonary disease; however, the role of the respiratory muscle pump in the dyspnea of chronic heart failure has not previously been examined. To assess respiratory muscle strength and its relation to dyspnea during daily activity, we measured maximum inspiratory and expiratory mouth pressures as indices of respiratory muscle strength and the baseline dyspnea index in nine stable, chronic cardiac pump failure patients who had no evidence of primary lung disease, and in nine age- and sex-matched healthy control subjects. The chronic heart failure patients, when compared with their matched control subjects, had reduced inspiratory and expiratory muscle strength, and both inspiratory and expiratory muscle strength were significantly correlated with dyspnea during daily activity (r2 = 0.80, p = 0.001 and r2 = 0.45, p = 0.05, respectively). Inspiratory muscle strength accounted for all of the variance in dyspnea that was correlated with respiratory muscle strength when the relative contributions of inspiratory and expiratory muscle strength were examined. There was no correlation between lung volumes or spirometry and dyspnea in the heart failure patients. These findings indicate that patients with stable chronic heart failure have inspiratory and expiratory muscle weakness and further suggest that the respiratory muscle pump significantly contributes to the dyspnea during the activities of daily living.


Study of bronchus-associated lymphoid tissue in patients with diffuse panbronchiolitis. Bronchus-associated lymphoid tissue (BALT) is thought to affect local immunologic defense mechanisms. Studies of BALT, however, have mainly been focused on animals, and information detailing the BALT structure and functions in humans is scanty. Therefore, the purpose of this study is to clarify BALT morphology and immunologic findings in patients with diffuse panbronchiolitis (DPB) and compare them with BALT findings in animals. Thus, in 17 patients diagnosed with DPB, open-lung biopsy specimens were reviewed, and lymphoid follicles with a lymphoepithelium that was determined as BALT were identified in 12 patients. BALT was found mostly at bifurcations from nonrespiratory bronchioles to respiratory bronchioles. This lymphoepithelium was devoid of cilia and was diffusely infiltrated with CD4-positive (helper/inducer) T cells. The majority of T cells were confined to the parafollicular area with a predominance of helper T cells. Also, a few high endothelial venules were recognized in the parafollicular area. Further, in a follicular area situated in the center of the BALT, a number of sigM-positive cells were found to be heavily distributed, suggesting that this follicular area has the characteristics of a B cell zone equivalent to a germinal center of the peripheral lymph nodes. These observations are comparable with BALT findings in animals and suggest that BALT is at least partially related to the local immune response.


Hypersensitivity pneumonitis in athymic nude mice. Additional evidence of T cell dependency. We previously demonstrated that C57 Black/6 mice develop lung lesions similar to human hypersensitivity pneumonitis (HP) by repeated transnasal administration of Thermoactinomyces vulgaris (Tv) antigen. To elucidate the role of T cells in the development of this disease, Tv antigen (90 micrograms/day) was transnasally administered to athymic nude C57 Black/nu/nu (nu) mice and their littermates (+/nu) three times a week for 3 wk. The nude mice developed minimal lung lesions, whereas their thymus-intact littermates (+/nu) showed changes equivalent to those in C57 Black/6. Changes in local inflammatory cell responses were evaluated by bronchoalveolar lavage, and increases in the numbers of lymphocytes and macrophages were significantly less severe in the nude mice than in the +/nu mice. Interestingly, the increase in polymorphonuclear leukocytes 6 h after the last antigen inoculation was equivalently seen in both groups. When spleen-derived T cells (more than 95% Thy-1.2+) from the sensitized +/nu mice were adoptively transferred to nude mice, the HP-like lesions in the recipients were found after Tv antigen challenge. These results suggest that Thy-1.2+ T cell-mediated immunity was necessary for the development of HP in this murine model.


Pulmonary complications in lymphoma patients treated with high-dose therapy autologous bone marrow transplantation. To define the incidence and spectrum of pulmonary complications following autologous bone marrow transplantation (BMT), we retrospectively reviewed the course of 77 consecutive patients with Hodgkin's disease (HD) and non-Hodgkin's lymphoma (NHL) who failed conventional therapy and underwent autologous BMT. Forty-five percent of the 77 patients developed respiratory complications with a mortality from pulmonary causes of 26%. A total of 38 episodes of respiratory compromise occurred in 35 patients. Infections accounted for 15 episodes (39%) and included bacterial (16%), Aspergillus (8%) cytomegalovirus (8%), Herpes simplex (3%), and other (5%) pneumonias. The spectrum of infections was similar to that reported following allogeneic BMT, but cytomegalovirus pneumonia was not as frequent a problem in those with autologous transplant. Mortality from pulmonary infections was 33%. Noninfectious disorders accounted for 23 episodes (61%) and included recurrent HD (18%), radiation/drug toxicity (16%), and acute respiratory failure thought secondary to pulmonary alveolar hemorrhage (26%). This latter entity developed acutely within 2 wk following BMT and was associated with use of thoracic radiation for treatment of malignant disease in the chest just prior to BMT (p < 0.05). It was not associated with the age of the patient or presence of thrombocytopenia, coagulopathy, renal insufficiency or neutropenia (p NS). Mortality from noninfectious causes was 65%, but in those with pulmonary hemorrhage it was 100%. In conclusion, pulmonary complications are a major source of morbidity and mortality in patients with HD and NHL undergoing autologous BMT.(ABSTRACT TRUNCATED AT 250 WORDS)


Disordered pathways of fibrin turnover in lung lavage of premature infants with respiratory distress syndrome. Premature infants who have self-limited respiratory distress syndrome (RDS) rapidly improve, whereas infants with a complicated respiratory course are more likely to develop bronchopulmonary dysplasia (BPD), a chronic lung disorder that is the result of prolonged lung injury and impaired healing. The balance of competing activities of coagulation and fibrinolysis may contribute to the premature lung's response to acute injury and determine, in part, whether there is early resolution or protracted alveolar inflammation. To determine the relative activities of the coagulation and fibrinolytic pathways in neonatal lung injury, procoagulant (PC) and plasminogen activator (PA) activities were measured in undiluted cell-free lung lavage samples obtained serially over the first 28 days of life from 11 infants with self-limited RDS, 11 infants with evolving BPD, and 5 mechanically ventilated control infants without lung disease. Lung lavage from all three groups contained readily detectable procoagulant activity due mainly to the tissue factor-Factor VII complex. Plasminogen activator activity was relatively high in control lavage samples but depressed on the first day of life in the two groups of infants with lung disease: median, 0.3814 IU/ml (control); 0.0541 IU/ml (RDS); and 0.0454 IU/ml (BPD), p < 0.05 in each case compared with control. Two infants with severe lung disease had no detectable plasminogen activator activity in lung lavage on the first day of life. Depressed fibrinolytic activity correlated with severity of lung disease assessed radiographically and by pulmonary function measurements. Plasminogen activator activity was due to both tissue plasminogen activator and urokinase.(ABSTRACT TRUNCATED AT 250 WORDS)


Identification of T lymphocytes, macrophages, and activated eosinophils in the bronchial mucosa in intrinsic asthma. Relationship to symptoms and bronchial responsiveness. Using immunohistochemistry and a panel of monoclonal antibodies, we have compared T-lymphocyte, eosinophil, macrophage, and neutrophil infiltration in bronchial biopsies from 10 intrinsic (nonallergic) asthmatics (IA) and seven extrinsic (allergic) asthmatic (EA), with similar degrees of disease severity. The results were compared with 12 normal healthy nonatopic controls (NC). All subjects were nonsmokers and were not taking oral or inhaled corticosteroids. An intense mononuclear cell infiltrate was identified in IA with an increase in the number of CD45+ cells (total leukocytes), CD3+ and CD4+ lymphocytes, and CD68+ macrophages (p < 0.03, p < 0.01, p < 0.03, and p < 0.03, respectively), compared with NC. Increases were also found in CD4+ (p < 0.05) and CD68+ (p < 0.05) cell numbers between IA and EA. IL-2 receptor-bearing cells (CD25+) and the number of total (MBP+) and actively secreting (EG2+) eosinophils, were also increased in IA compared with NC (p < 0.01, p < 0.01, and p < 0.01, respectively). Similar increases in EG2+ eosinophils and CD25+ (IL-2 receptor-positive) cells were observed in EA (p < 0.01 and p < 0.02, respectively). No differences were detected in the three groups for the number of elastase-positive cells (neutrophils). EG2+ numbers in IA correlated with the Aas asthma symptoms score (r = 0.65, p < 0.05), whereas EG2+ cell numbers in all asthmatics (IA + EA) correlated with airway methacholine responsiveness (r = -0.55, p < 0.03) and with the Aas asthma symptom score (r = 0.54, p < 0.03).(ABSTRACT TRUNCATED AT 250 WORDS)


Airway inflammation and occurrence of delayed bronchoconstriction in exercise-induced asthma. We studied nine asthmatic patients with a history of exercise-induced asthma (EIA) in order to investigate whether inflammatory changes in the airways occur after exercise and are eventually associated with the development of a late-phase asthmatic response. On two separate study days, bronchoalveolar lavage (BAL), bronchial lavage (BL), and bronchial biopsy (BB) were performed 3 h after an exercise or a methacholine (MCh) challenge. On two other separate occasions, FEV1 was monitored for 12 h after identical exercise and MCh challenges not followed by BAL, BL, and BB. We found a greater percentage of eosinophils in BAL after exercise versus MCh challenge (p < 0.05). In five patients, BAL eosinophils after exercise were > or = 2% of total inflammatory and immunoeffector cells. In three of these patients an FEV1 fall > 20% of control was recorded 5 to 12 h after exercise. However, two of these patients had 2% or more eosinophils in BAL and similar late falls of FEV1 after MCh challenge. The percentage of degranulating mast cells in BB was higher (p < 0.05) after EIA than after MCh, but no significant differences were found in BL histamine and leukotrienes. We conclude that (1) exercise may enhance mast cell degranulation and eosinophilic inflammation of the airways, and (2) a delayed bronchoconstriction after exercise is not specific to EIA but is more likely the result of fluctuations in lung function associated with airway inflammation.


Bilateral bronchial anomaly. A pathogenetic factor in spontaneous pneumothorax. Flexible fiberoptic bronchoscopy (FFB) was performed in 26 never-smokers with healed spontaneous pneumothorax (SP) with the aim of detecting and localizing any bronchial obstruction, including congenital anomaly. In a case-control study these patients were compared with a consecutive, randomly sampled, control group of 41 patients who were undergoing FFB for respiratory symptoms but who had not had a SP. In both groups the endobronchial anatomy with respect to such anomalies, which could be classified as disproportionate bronchial anatomy, an accessory bronchus, and/or a missing bronchus, was compared with normal anatomy. All except one of the 26 patients with SP but only four of the 41 control patients without SP had such bronchial anomalies bilaterally. This corresponds to an odds ratio of 231 (95% confidence interval, 24 to 880; p < 0.001). The significantly higher frequency of bilateral bronchial anomalies in never-smokers with SP suggests that a virtual prerequisite for the occurrence of SP has been found. However causal links between the probably congenital bronchial anomalies and SP have not yet been identified.


Aerosolized lidocaine reduces dose of topical anesthetic for bronchoscopy. Conventional aerosol techniques were used to determine if inhalation of lidocaine can supplement topical anesthesia applied during bronchoscopy. Aerosols of either saline or lidocaine (50 mg at either 2 or 4% concentrations) were generated by jet nebulizer and administered with or without intermittent positive-pressure breathing. Patients (n = 38) after aerosol inhalation were administered 2% lidocaine (atomized and instilled) for suppression of the gag reflex, control of cough, and airway anesthesia. For five of the patients, prior to bronchoscopy, additional studies with radioaerosols and scintillation scans were accomplished with the same aerosol methodology to demonstrate lung distribution of deposited aerosol. For five patients who received 2% lidocaine aerosol prior to bronchoscopy, the subsequent topical dose of anesthetic required for the procedure was 186 +/- 34 (SEM) mg lidocaine. Nine patients in a control group received saline aerosol and required significantly more anesthetic, i.e., 308 +/- 26 mg; procedures were completed on average within 50 min. The largest difference was in the amount delivered to the upper airway (naris, pharynx, epiglottis, and larynx), i.e., 144 +/- 26 mg for saline control versus 48 +/- 16 mg for lidocaine aerosol protocol. Airways distal to the cords required less anesthesia also, on average, 77 mg for the saline control versus 46 mg for the lidocaine aerosol protocol (p < 0.05). Topical anesthetic dosage data were replicated in 12 additional patients studied by a different bronchoscopist. No additional benefit was afforded by premedication with 4% lidocaine aerosol rather than the 2% aerosol (n = 12). We conclude that aerosol modalities can supplement topical anesthesia during bronchoscopy, primarily by reducing the dose required to anesthetize the upper airway.


Polymicrobial bacterial pericarditis after transbronchial needle aspiration. Case report with an investigation on the risk of bacterial contamination during fiberoptic bronchoscopy. A 63-yr-old man developed pericardial effusion with tamponade after transbronchial needle aspiration (TBNA) of a subcarinal mass. A diagnosis of polymicrobial bacterial pericarditis was made when pericardiocentesis revealed purulent fluid that grew a mixed culture of anaerobes and aerobes, organisms that constitute part of the normal upper respiratory tract flora. To examine the possibility that contamination of the transbronchial needle (TBN) could lead to purulent pericarditis by inoculation of bacteria into the mediastinum, quantitative cultures of the TBN content were performed in seven consecutive patients. Abundant growth of multiple anaerobic and aerobic organisms was demonstrated in all seven cultures. We conclude that subcarinal TBNA is another potential cause of purulent pericarditis. This results from upper respiratory tract contamination of the open distal end of the TBN as it passes through the suction channel of the bronchoscope.


Severe hypoxemia-associated intrapulmonary shunt in a patient with chronic liver disease: improvement after medical treatment. A 20-yr-old woman with chronic liver disease and angioimmunoblastic lymphadenopathy presented with marked hypoxemia caused by intrapulmonary shunt. Her respiratory tract showed her to be free of angioimmunoblastic lymphadenopathy manifestations. After 12 months of treatment with cyclophosphamide and corticosteroid, the immunologic disease disappeared. Unexpectedly, hypoxia-associated intrapulmonary shunt was no longer present either. To the best of our knowledge, this is the first case of dramatic improvement, with medical therapy, of severe hypoxemia related to noncirrhotic liver disease. However, the mechanism by which this treatment caused the regression of intrapulmonary shunt is unknown.


NHLBI workshop summary. Research needs and opportunities related to respiratory health of women. To summarize, the overwhelming conclusion of the participants was that relatively little is known about respiratory health and disease in women. Much work will be required to obtain even the most basic information upon which hypotheses for future research can be developed and to determine whether pulmonary biology differs between men and women and between women of various ages. The many recommendations and suggestions generated at the workshop should serve to underscore the unique opportunities for research in the field of respiratory health of women.


Measurement of sialic acid in serum and urine: clinical applications and limitations. Many recent studies have examined the sialic acid content of serum or urine in various pathological states. We have briefly reviewed the substances which contribute to the observed total sialic acid concentration, and given an overview of assay methods used. Three major areas of clinical interest in sialic acid metabolism are discussed. Serum total sialic acid, 'lipid-bound' and 'protein bound' sialic acid have all been proposed as tumour markers; but the usefulness of any of these tests is severely limited by changes due to accompanying inflammatory processes. Serum total sialic acid is not a valuable simple marker of an acute phase response. Urinary free and bound sialic acid measurements should be included in screening protocols for inherited disorders of lysosomal metabolism. Current developments in research and potential applications within the clinical biochemistry laboratory are briefly discussed.


Enzyme activities of the mitochondrial energy generating system in skeletal muscle tissue of preterm and fullterm neonates. Quadriceps muscle specimens from autopsy of 28 neonates (gestational age 25-42 weeks) were investigated to determine pyruvate and malate oxidation rates and several enzymes of the mitochondrial oxidative process. In general, the levels of all mitochondrial parameters measured, including carnitine levels, were lower in the neonates who died within the first week of life than those in the control group (age > 5 years). Pyruvate and malate oxidation rates (P < 0.05), activities of pyruvate dehydrogenase complex (P < 0.10) and succinate: cytochrome c oxidoreductase (P < 0.05) increased significantly with gestational age. Pyruvate oxidation rates (P < 0.05) as well as activities of citrate synthase (P < 0.05) and NADH:Q1 oxidoreductase (P < 0.05) were significantly lower in the group of very preterm infants at an age of 1-7 days compared with very preterm infants at an age between 3-8 weeks. We conclude from our study that special reference values are necessary for a correct biochemical diagnosis of mitochondrial encephalomyopathies in the neonatal period. Differences between preterm and fullterm children of the same age (1 week) indicate a maturational process in human muscle tissue during gestation. Comparison of two different age groups within the very preterm neonates point to a postnatal maturation of the mitochondrial energy metabolism, at least in preterm neonates.


Urinary dopamine excretion in chronic renal disease. We have measured unconjugated urinary dopamine and investigated its relationship to glomerular filtration rate in two groups of 22 patients with chronic renal failure matched for age and sex, one group with primary glomerular disease, the other with tubulo-interstitial disease. Urine dopamine excretion was similar in both glomerular and tubulo-interstitial disease groups, and correlated significantly with creatinine clearance. Although urinary protein excretion was significantly higher in glomerular disease, urinary sodium excretion, fractional sodium excretion, urine flow rate and free water clearance were similar in both groups and did not correlate with dopamine excretion. These results suggest that in patients with chronic renal disease, urinary dopamine excretion is mainly under the influence of the glomerular filtration rate, irrespective of the underlying pathology, and therefore interpretation of urinary dopamine excretion requires a knowledge of the patients' renal function.


Protein measurement of particulate and solubilized ovine liver membranes. Protein concentration measurements are critical in biochemical work with cellular membranes, including the determination of cell surface receptor concentration in human malignant tissues obtained at surgery or after biopsy. In this study we compared the results of protein concentration measurements in ovine liver cellular membranes using either particulate preparations or membranes solubilized with four different detergents. In all cases protein was determined by two different indirect methods (Lowry's Folin phenol method and Bradford's Coomassie Brilliant Blue dye binding method) and compared to the direct biuret method. Our results indicate that the direct biuret method gives the highest protein concentrations followed by the method of Lowry. Maximal concentrations (approaching those obtained by the direct biuret method) were obtained after membrane solubilization with Triton X-100 (3-5%). It is suggested that either the direct biuret method (whenever protein concentrations permit it) or the method of Lowry after solubilization of membranes with Triton X-100 (3-5%) should be used preferentially for the determination of membrane protein samples.


Improved flow cytometric method for HLA-B27 typing. HLA-B27 is a cell marker of clinical interest because of its high association with certain diseases. The HLA-B27 antigen was detected on lymphocytes using a monoclonal antibody in an indirect immuno-fluorescence assay using a fluorescence flow cytometer. The considerable crossreaction of the monoclonal antibody with the HLA-B7 antigen was effectively suppressed by masking it by means of human anti-HLA-B7 antiserum. The flow cytometric method was evaluated by comparing the results with those obtained by the standard lymphocytotoxicity test and showed complete agreement in 107 selected patient samples.


Glycine absorption from irrigation fluid during endoscopic resection of rectal tumours. A significant rise in plasma glycine concentration was observed in nine elderly patients undergoing endoscopic transanal resection of rectal tumours using glycine solution for irrigation. Despite the obvious absorption of glycine, plasma sodium concentration and osmolality were not significantly altered, except in a patient whose rectal wall was perforated. In this case plasma sodium and osmolality fell, and the patient developed hyperammonaemia. Potentially adverse metabolic consequences may occur when there are surgical complications, but in the absence of such problems the absorption of glycine appears to cause little effect.


[Encrusted pyelo-ureteritis].

[Encrusted pyelo-ureteritis]. The authors report a case of right encrusted pyelo-ureteritis with no other sites, either in the bladder or on the other side. They discuss possible theories of pathogenesis of the encrusted plaques, and question how far corynebacterium, group D2, could be concerned in the genesis of such lesions.


[Isolated retrovesical and retroperitoneal hydatid cysts. 5 case reports]. In the light of five cases of isolated retrovesical and retroperitoneal hydatid cysts, the characteristics of these rare locations are reviewed and the hypotheses of pathogenesis are discussed. An abdominal or abdomino-pelvic mass is the most frequent sign. Ultrasonography and computed tomography are valuable in the diagnosis of these lesions. Partial cysto-pericystectomy via an extraperitoneal approach is the treatment of choice for these hydatid cysts.


[Calcified necrosing cystitis].

[Calcified necrosing cystitis]. Calcified necrotising cystitis is an urological curiosity, the authors report two cases. With subsidence of the acute process, regeneration and fibrosis take place, with contracture of the bladder and reduction of its capacity. Augmentation ileocystoplasty was performed with success in one case. The clinical and therapeutic aspects of this rare disease are discussed.


[Traumatic rupture of the corpus cavernosum. 25 case reports and literature review]. On the basis of a series of twenty-five cases of traumatic rupture of the corpus cavernosum and a review of literature, the characteristics of this rare affection are reviewed. This injury typically occurs in young adults. The most frequent mechanism is rough manipulation of the erect penis. Urethral injury is associated in 10 per cent of patients. The characteristic history and physical findings usually yield the diagnosis. The preferred form of management is early surgical repair of the tunical defect, by distal circumferential incision. Post-operative complications are rare (8 per cent). The treatment of associated urethral injuries and late complications resulting from untreated rupture is reviewed.


[A case report of epididymo-testicular bilharziasis in a child]. The disease was revealed by a tumor of the left testis. Ultrasound showed an enlargement of testis and epididymis with some hypoechogenic areas resembling epididymo-orchitis or tumor. The location of bilharzial lesions in the testis is very rare. The main sign of urogenital bilharziosis, hematuria, was present. Orchidectomy is the treatment of choice when the testis is destroyed and only histological study is able to confirm the nature of testicular mass.


[Evolution and treatment of 8 patients with Fournier's perineal-scrotal gangrene]. The authors present eight cases of Fournier's gangrene treated from 1987 to 1989 (3 years). There was no discernable cause for 5 patients (62.5%). Immediate aggressive surgical debridement of all necrotic tissue was performed. Intravenous antibiotics and resuscitation fluid and hyperbaric oxygenation were also administered to all patients. Skin grafting was performed for 4 patients (40%), 3 weeks after surgical debridement. One patient died (12.5%) and 2 subsequently developed an urethral stricture. Etiology, treatment and outcome of Fournier's gangrene are discussed and the results of the study are compared to other studies. This disease is still serious, its pathogenesis is not completely elucidated, but when immediate adequate treatment is performed, the outcome is generally favourable.


In vitro activity of RU 29246, the active compound of the cephalosporin prodrug ester HR 916. The in vitro activity of RU 29246 was compared with those of other agents against 536 recent clinical isolates. The MICs of RU 29246 for 90% of members of the family Enterobacteriaceae tested (MIC90s) were less than 2 micrograms/ml except those for Morganella spp. (16 micrograms/ml) and Proteus spp. (8 micrograms/ml). RU 29246 was active against Staphylococcus aureus (MIC90, < or = 8 micrograms/ml) and against Staphylococcus saprophyticus and coagulase-negative staphylococci (MIC90s, < or = 2 micrograms/ml). Streptococci and Neisseria gonorrhoeae were highly susceptible to RU 29246, and the activity of the agent against isolates of Streptococcus pneumoniae (MIC90, < or = 0.5 micrograms/ml), Haemophilus influenzae (MIC90, < or = 2 micrograms/ml), and Moraxella catarrhalis (MIC90, < or = 2 micrograms/ml) was comparable to those of the other cephalosporins tested. RU 29246 was insusceptible to hydrolysis by the common plasmid-mediated beta-lactamases (TEM-1 and SHV-1). However, hydrolysis by the new extended-spectrum beta-lactamases (TEM-3, TEM-5, and TEM-9) was detected. Results of the study suggested that RU 29246 should be investigated clinically for use in the treatment of a wide range of infections.