Cloning of a yolk protein gene family from Caenorhabditis elegans.

We have cloned a family of five genes which encode the 170,000 Mr yolk proteins in the nematode Caenorhabditis elegans. The genes and their messenger RNAs are about 5 X 10(3) base-pairs in length. Thus most of the length of each gene is exon, although a few small introns have been discovered. Based on hybridization and restriction mapping experiments, the genes can be subdivided into two subfamilies: YP1-YP2 and YP3-YP4-YP5. Within a subfamily the genes are nearly identical. While most of the genes are not clustered, YP3 and YP4 are tandemly linked. Hybrid-arrest translation experiments demonstrate that the YP3-YP4-YP5 subfamily encodes the yp170A yolk protein, while the YP1-YP2 subfamily encodes the yp170B yolk protein. RNAs homologous to these genes are abundant in the adult hermaphrodite, but missing from larvae and males. Furthermore, RNA isolated from dissected intestines is highly enriched for sequences that hybridize to the genes, whereas RNA from gonad or body wall is nearly devoid of these sequences. Thus, this gene family is apparently expressed only in the intestine of the adult hermaphrodite.

Enhanced adhesion of polymorphonuclear leukocytes to anoxic cultured vascular endothelium.

This study showed that the adherence of human polymorphonuclear leukocytes (PMN) to monolayer cultures of human umbilical vein endothelial cells (HUVEC) was increased when the latter were rendered anoxic. This adhesion was greater after 4-5 h than after 8 h of anoxia, but even at 8 h was significantly above the level of adhesion to HUVEC maintained under normoxic conditions for the same period. The changes in adhesion were not dependent on the viability of HUVEC during anoxia. Anoxia-induced adhesion was prevented by addition of cycloheximide (5 micrograms/ml) to the cultures, suggesting that it depended on HUVEC protein synthesis. Enhanced adhesion was also prevented by addition of a monoclonal antibody directed against the cytokine, interleukin-1 alpha (IL-1 alpha). These findings are consistent with a role of endogenous IL-1 alpha as a mediator of the anoxia-induced adhesion of PMN to HUVEC.

Changes in pulmonary function test results after 1 year of therapy as predictors of survival in patients with idiopathic pulmonary fibrosis.

The study group consisted of 58 patients with idiopathic pulmonary fibrosis (IPF) recognized between 1970 and 1991 who were treated for their pulmonary disease, survived for at least 1 year from the time of initiation of treatment, and had forced vital capacity (FVC) measurements at the time of diagnosis and 9 to 15 months later. Forty-four of the patients also had a single-breath diffusing capacity (Dsb) measured initially and after 9 to 15 months of treatment and 33 patients had an arterial blood gas, breathing room air at the time of diagnosis and 9 to 15 months into therapy. Patients' conditions were classified as improved, unchanged, or worse after the year of treatment based on each of the three pulmonary function tests. A > or = 10% increase in FVC, > or = 20% increase in Dsb, and > or = 5 mm Hg decrease in alveolar-arterial difference in oxygen partial pressure [P(A-a)O2] defined improved function. A > or = 10% decrease in FVC, > or = 20% decrease in Dsb, and > or = 5 mm Hg increase in P(A-a)O2 defined worse function. Patients with < 10% change in FVC, < 20% change in Dsb, and < 5 mm Hg change in P(A-a)O2 were regarded as having unchanged conditions. Kaplan-Meier survival plots and the Cox proportional hazard regression model were used to analyze survival time in relation to change in pulmonary function after 1 year of therapy. Patients with an improved or unchanged FVC at 1 year had no difference in survival (p = 0.75), but both showed enhanced survival compared with patients with a > or = 10% reduction in FVC with 1 year of treatment (p < 0.001). Patients with an improved or unchanged Dsb at 1 year also had no difference in survival (p = 0.21) but again, both showed enhanced survival compared with patients with > or = 20% decrease in Dsb with 1 year of treatment (p < 0.001). Changes in gas exchange after 1 year of treatment did not correlate with survival in the three groups. There was a trend for longer survival in improved patients compared with those with worsening gas exchange, but the p value was not significant at 0.17. We conclude that changes in the FVC and Dsb after 1 year of treatment are strongly predictive of duration of survival in patients with IPF.

A clinical profile of chronic bacterial pneumonia. Report of 115 cases.

OBJECTIVE: To review the clinical presentation, radiology, microbiology, and response to therapy of patients with chronic bacterial pneumonia. DESIGN: A retrospective analysis. SETTING: An urban tertiary care medical center. PARTICIPANTS: One hundred fifteen patients with pulmonary and/or constitutional symptoms of at least 1 month's duration with 4,000 or more colony-forming units (CFUs) of a single bacterial species identified by quantitative culture obtained via fiberoptic bronchoscopy. MEASUREMENTS: Charts were analyzed for presence or absence of any predisposing illness, symptoms at presentation, roentgenographic abnormalities, microbiologic results, findings at fiberoptic bronchoscopy, and results of therapeutic intervention. RESULTS: Sixty-five percent of patients with chronic bacterial pneumonia had a predisposing disease, 35 percent were "normal." Cough, fatigue, dyspnea, and weight loss were predominant symptoms in both groups. Bronchogenic carcinoma was newly diagnosed in 16 patients (14 percent). Haemophilus influenzae or alpha-hemolytic streptococcus was isolated in 68 percent of patients. Risk of recurrence of infection was inversely associated with duration of therapy in both groups. CONCLUSIONS: Chronic bacterial pneumonia is more common than previously recognized. It occurs in patients with and without a predisposing illness. Clinical presentation, roentgenographic appearance, and bacteriology are similar between the two groups. Cure requires prolonged antibiotic therapy.

The diagnosis of ventilator-associated pneumonia: a comparison of histologic, microbiologic, and clinical criteria.

STUDY OBJECTIVE: To evaluate histologic, microbiological, and clinical criteria in the recognition of ventilator-associated pneumonia (VAP) in patients who died while mechanically ventilated. METHODS: The study group consisted of 39 patients who died after a mean of 14 days of mechanical ventilation. Postmortem fiberoptic bronchoscopy (FOB) and open lung biopsy were performed with collection of specimens initiated <1 h after death. The microbiological specimens included suction catheter aspirate of tracheal secretions, FOB-guided protected specimen brush (PSB) of tracheal secretions, blindly placed PSB in a distal airway, FOB-guided PSB in a distal airway, and FOB-guided BAL fluid (BALF) in a distal airway. Qualitative bacteriologic study was performed on all specimens, and quantitative bacteriologic study was performed on all but the suction catheter aspirate of the trachea. A biopsy specimen of peripheral lung parenchyma from the same region sampled by FOB was sent for quantitative culture and histologic analysis. The BALF was analyzed for cell population and percent of neutrophils containing intracellular organisms. The clinical criteria selected for comparison with histologic and microbiological results included a temperature > or =38.5 degrees C during the 48 h prior to death, a WBC count > or =15,000/mm3 in the 48 h prior to death, presence of a bacterial or fungal pathogen on the last sputum culture, radiographic worsening in the week prior to death, and worsening gas exchange defined as a 15% decrease in the PaO2/fraction of inspired oxygen ratio in the 72 h prior to death. RESULTS: None of the quantitative cultures had a reliable positive predictive value for histologic pneumonia. None of the five clinical criteria tested showed agreement with the presence or absence of histologic pneumonia. There was a significant correlation between qualitative and quantitative microbiological results from the distal airway/FOB-guided PSB, distal airway/BALF, and quantitative culture of the lung parenchyma. Also, suction catheter aspirate of the trachea had a sensitivity of 87% in recognizing the bacterial species simultaneously present in lung parenchyma. None of the patients with histologic pneumonia had <50% neutrophils in the BALF. CONCLUSIONS: Neither the bacterial, density from the four airway quantitative cultures, nor the bacterial density from quantitative culture of lung parenchyma accurately separated the histologic pneumonia and nonpneumonia groups. No clinical criteria or combination of clinical criteria correlated with the presence or absence of histologic pneumonia. A BALF with <50% neutrophils had a 100% negative predictive value for histologic pneumonia. A BALF quantitative culture had a sensitivity of 63%, specificity of 96%, and positive predictive value of 91% in recognizing sterile lung parenchyma. Thus, BALF may have a role in excluding pneumonia/infection in the ventilated patient. Antibiotic choice for the empiric therapy of VAP can be accurately guided by the microbial population recognized through culture of a tracheal suction catheter aspirate.

Reproducibility of the histologic diagnosis of pneumonia among a panel of four pathologists: analysis of a gold standard.

STUDY OBJECTIVE: To establish a histologic diagnosis of pneumonia by consensus of a panel of pathologists, to test the interobserver and intraobserver variation in the histologic diagnosis of pneumonia, to compare the diagnostic accuracy of diagnosing pneumonia with and without preselected histologic criteria, and to establish more specific histologic criteria for the diagnosis of pneumonia. METHODS: The study group consisted of 39 patients who died after a mean of 14 days of mechanical ventilation. A postmortem open lung biopsy was performed on all patients. The tissue was reviewed independently by four pathologists who categorized the slides from each patient as showing or not showing pneumonia. Interobserver variation was calculated using the kappa statistic. Six months following the initial evaluation, the same slides were resubmitted to one of the pathologists for reevaluation to look for intraobserver error. Finally, the slides were reviewed and categorized by the criteria of Johanson et al into no pneumonia, mild, moderate, or severe bronchopneumonia. A comparison was made of the patients selected as demonstrating histologic pneumonia by each of the examinations. RESULTS: The reliability coefficient (kappa) measuring agreement among the four pathologists was good at 0.916. However, the prevalence of pneumonia as determined by each of the four pathologists varied; pathologist A, 15 of 39 (38%); pathologist B, 12 of 39 (31%); pathologist C, 9 of 39 (23%); and pathologist D, 7 of 39 (18%). Resubmitting the same slides to the same pathologist 6 months later resulted in reclassification of 2 of 39 patients. Using the histologic criteria of Johanson and colleagues, 14 patients were selected as having pneumonia compared with only nine patients selected by consensus of three of four pathologists. CONCLUSIONS: Recognition of histologic pneumonia varies among pathologists. The preselected criteria of Johanson and colleagues detected histologic pneumonia in eight of nine patients picked by consensus of pathologists, but six additional patients classified as "no histologic pneumonia" by the consensus of pathologists were judged to have histologic pneumonia by these criteria. The results established the necessity for standardization of histologic criteria for studies using biopsy as the gold standard for bacterial pneumonia. An atlas showing the criteria used in our selection was developed.

Central cardiovascular effects of mammalian neurohypophyseal peptides in conscious rats.

To confirm and extend the results of previous studies which demonstrated central cardiovascular effects of vasopressin in anesthetized rats, we determined blood pressure and heart rate changes for 30 minutes after intracerebroventricular injections of arginine vasopressin, arginine vasotocin and oxytocin in conscious rats. As compared to sham injections, significantly greater increases in either systolic or diastolic blood pressure were noted over the 30 minutes which followed the injection of 0.15, 1.0 or 10.0 nM of either vasopressin or vasotocin. In animals given vasopressin, plasma levels of the peptide were determined. There was a substantial increase in plasma vasopressin only after the highest dose. Overall blood pressure responses to doses of oxytocin as high as 100 nM were not significantly different than sham injections. Heart rate following both vasopressin and vasotocin was increased at 0.15 nM, was initially decreased then increased at 1.0 nM and was substantially decreased after the 10.0 nM dose. There was a significant increase in heart rate at the 10.0 nM and 100 nM doses of oxytocin. Dose response curves for systolic blood pressure and heart rate 20 minutes after injection were similar for vasopressin and vasotocin. We conclude that arginine vasopressin has significant central pressor and tachycardic effects in conscious rats, and it is related, at least in part, to the tail structure of the peptide, which is shared with arginine vasotocin.

Prostaglandin E1: a review.

Work on the structure of prostaglandin E1 (PGE1), isolated from natural sources, was completed 25 years ago (1). Shortly after, methods for the chemical synthesis of PG with their natural configuration were developed in the laboratories of the UpJohn Company (2) and of E. J. Corey (3) and, by the late sixties, PGE1 became widely available. The information since accumulated about its biological and clinical effects is more substantial than for any other PG. This review will draw together some of this information, focusing on recent studies of its mechanisms of action.

Slowly resolving, chronic, and recurrent pneumonia.

Slowly resolving, chronic, and recurrent pneumonias are clinical patterns that are often misunderstood and mistreated. This article reviews the natural history, clinical presentation, roentgenography, bacteriology, and relation to underlying illnesses of each of these atypical forms of community-acquired pneumonia. Standard definitions are presented and provide the physician with a means to classify the pneumonias. Appropriate diagnostic evaluation and therapy are also discussed.

Treatment with corticosteroids.

Oral corticosteroids remain the cornerstone therapy for sarcoidosis. Critical clinical decisions include selecting the patient who should be treated, dose and duration of therapy, and accurate analysis of the anticipated benefits and potential side effects for the individual patient. The treatment of pulmonary and cardiac sarcoidosis is emphasized and the role of inhaled corticosteroids in the treatment of pulmonary sarcoidosis is reviewed.

A randomized study of a pH sensor feeding tube vs a standard feeding tube in patients requiring enteral nutrition.

Postpyloric feedings are a widely practiced form of enteral nutrition. We prospectively randomized two groups of hospital patients to receive a standard feeding tube or a feeding tube that uses a pH sensor to facilitate postpyloric placement and compared placement speed and accuracy, displacement detection, and costs for the two groups. Thirty-nine patients were randomized, with 20 receiving a pH sensor feeding tube and 19 an identical non-pH sensor feeding tube. An x-ray of the kidneys, ureter, and bladder was taken at 1, 6, and 48 hours after placement in both groups. Separate cost-benefit analyses were done by using retrospective chart review of costs for a separate 20-patient standard feeding tube group and calculated costs for a 20-patient hypothetical pH sensor group. At 1 hour, the duodenum was reached in 53% of the pH sensor feeding tube patients and 45% of the standard feeding tube patients (the difference was not significant). At 48 hours, 93% of the pH sensor feeding tubes reached the duodenum vs 67% of the standard feeding tubes (p < .08). Thirty percent of the pH sensor patients had an initial gastric pH > or = 4, negating pH sensor benefit in tube placement. In the remaining 70% of the patients, placement with the pH sensor had a 100% specificity compared with the x-ray of the kidneys, ureter, and bladder. Displacement was easily detected with routine pH monitoring in three of the pH sensor feeding tube patients and corrected. It was detected in two standard feeding tube patients, one of whom aspirated.(ABSTRACT TRUNCATED AT 250 WORDS)

Dihomo-gamma-linolenic acid reverses hypertension induced in rats by diets rich in saturated fat.

This study has shown that hypertension induced in rats by a diet rich in saturated fat (16% coconut oil, 4% palmitic acid by weight) is reversed by the addition of the essential fatty acid, dihomo-gamma-linolenic acid (DHLA), at 5.0% but not at 0.5% of dietary energy. This potent effect of DHLA has been attributed to modulation of prostaglandin biosynthesis.

The metabolism of dihomo-gamma-linolenic acid in man.

Orally administered dihomo-gamma-linolenic acid (DHLA) is well absorbed in man; it appears in blood after ca. 4 hr first as triglyceride ester and later as phospholipid. After sustained-dosing, DHLA penetrated membrane pools and all phospholipid components but, depending on the dosage, reached a metabolic equilibrium in 4-16 days. Intact platelets do not accumulate arachidonate following DHLA administration, and species differences occur in the capacity of animals to metabolize DHLA to arachidonic acid (AA). The rat appears to be unusual in having a very active hepatic delta5-desaturase enzyme system. Potentially antithrombotic changes in platelet function which followed the administration of DHLA to man were accompanied by a significant increase in the capacity of platelets to synthesize PGE1. Concomitant increases in PGE2 synthesis do not apparently result from an increased production of AA and suggest that DHLA, or a DHLA metabolite, interferes with the metabolism of AA. Effects on thromboxane and prostacyclin synthesis are being studied.

Chronic pneumonias.

Some patients present in the outpatient setting with persistent respiratory and constitutional symptoms in association with unresolving parenchymal infiltrates on the chest radiograph. Clinicians must entertain a wide differential diagnosis as many infectious and noninfectious conditions can lead to this chronic pneumonia syndrome. The clinical presentation and radiographic abnormalities are often nonspecific. A significant proportion of patients have no underlying predisposing illness, and most of the bacterial pathogens are often considered constituents of the "normal respiratory flora." Accurate diagnosis generally requires bronchoscopic evaluation. Prolonged therapy is essential. This article reviews the epidemiology and the predominant bacterial, mycobacterial, and fungal pathogens associated with this syndrome. A brief discussion of some of the noninfectious processes is also included.

Clinical pathways: improving patient education and influencing readmission rates.

Clinical pathways have long been used as a mechanism for implementing a managed care delivery system. They have been effective in highlighting lengths of stay, outcome variances, and systems problems. Clinical pathways play an important part in enhancing quality improvement activities, especially in accordance with the standards of the Joint Commission on Accreditation of Healthcare Organizations. This article addresses benefits of clinical pathways in general, and benefits at the authors' facility in particular, specifically, a clinical pathway's effectiveness in influencing readmissions of patients with atrial fibrillation.

The effect of dihomogammalinolenic acid on platelet aggregation and prostaglandin release, erythrocyte membrane fatty acids and serum lipids: evidence for defects in PGE1 synthesis and delta 5-desaturase activity in insulin-dependent diabetics.

The effect of dihomogammalinolenic acid (DHLA) administration on platelet aggregation and prostaglandin production, erythrocyte fatty acid composition and serum lipids was compared in healthy subjects and insulin-dependent diabetics (IDDs). In healthy subjects, DHLA caused a significant inhibition of ADP-induced platelet aggregation and an increase in platelet PGE1 release; IDDs did not show these changes. There were no differences, however, in platelet thromboxane A2 (TXA2) or PGE2 release between healthy subjects and IDDs before and after DHLA. Following DHLA, the arachidonic acid content of erythrocytes increased in healthy subjects; this increase was not observed in IDDs. DHLA induced a significant fall in serum non-esterified fatty acid concentrations in both groups without altering either cholesterol or triglyceride concentrations. These data show for the first time that IDD platelets may have a specific defect of PGE1 synthesis quite distinct from the delta 5- and delta 6-desaturase defects known to be associated with experimental diabetes; this defect may contribute to platelet hyper-aggregability in diabetes; and DHLA has a potent antilipolytic effect in vivo; and erythrocytes from IDDs may have a delta 6-desaturase defect.

The C. elegans vitellogenin genes: short sequence repeats in the promoter regions and homology to the vertebrate genes.

The nematode Caenorhabditis elegans contains a small family of vitellogenin genes which is expressed abundantly, but only in the intestine of the adult hermaphrodite worm. In order to identify possible regulatory elements, we have sequenced the DNA surrounding the 5' ends of five of the six genes. Contained within regions which have largely diverged from one another, two different heptameric sequences are found repeated within the first 200 bp upstream of each of the genes. The first sequence, TGTCAAT, is present as a perfect heptamer at least once upstream of each gene. It is repeated in both orientations four to six times in each 5' flanking region, allowing a one-base mismatch. The second sequence, CTGATAA, is also present as a perfect heptamer in a restricted region upstream of each gene. These two sequence elements may be involved in regulation of the vitellogenin genes. Remarkably, the CTGATAA sequence is present in a similar location in the promoter regions of vertebrate vitellogenin genes. In fact, our data reveal a surprising degree of similarity between the nematode and vertebrate vitellogenins.

Chronic unreduced dislocations of the glenohumeral joint: imaging strategy and pathologic correlation.

Although the evaluation of acute or recurrent dislocations of the glenohumeral joint (GHJ) using routine radiography, specialized views, arthrography, and computed tomography (CT) has been extensively studied, methods of evaluation and osseous pathology of chronic unreduced GHJ dislocations are not well documented. In order to define such methods and pathology, we studied nine patients with chronic (greater than 6 months) unreduced anterior, posterior, superior, or central dislocations as well as one patient with a chronic rotator cuff injury; and seven scapular specimens, six with matching humeral specimens, with osseous abnormalities similar to those in the patients. We examined 15 additional scapular specimens with evidence of subluxation secondary to chronic rotator cuff injury. Standard, reformatted, and three-dimensional (3D) CT techniques were used to evaluate both the patients and the specimens. Our results indicate that characteristic alterations occur in the scapula and humeral head that are far more extensive than the classic Hill-Sachs, Bankart, and trough lesions. Such alterations occur at sites of osseous contact between the malpositioned humerus and scapula and are modified owing to prolonged adjustments made by the patients in an attempt to increase GHJ motion. Although routine radiography and specialized views are of some value, standard and 3D CT provide a more accurate analysis of the location and extent of the bone abnormalities and, as such, should be employed, particularly in the evaluation of patients in whom surgery is being considered.