1-ether-linked phosphoglycerides. Major endogenous sources of arachidonate in the human neutrophil.

This study has quantitated changes in the content of labeled and unlabeled arachidonate of neutrophil phosphoglyceride classes and subclasses during cell activation with ionophore A23187. The predominant pools of endogenous arachidonate in the resting neutrophil were found in ethanolamine (68%)-, choline (19%)-, and inositol (12.0%)-containing glycerolipids. Upon stimulation, endogenous arachidonate was lost from primarily ethanolamine (PE) greater than choline (PC) greater than inositol (PI)-linked phosphoglycerides. Released leukotriene B4 and 20-hydroxyleukotriene B4 accounted for 10-35% of the total arachidonate lost from all phosphoglyceride classes. In contrast to the mass loss, ionophore induced a decrease of labeled arachidonate from primarily PC and PI. In the resting neutrophil, 66% of the total arachidonate in PC was found in the 1-alkyl-linked fraction. Furthermore, loss of endogenous arachidonate from 1-alkyl-2-arachidonoyl sn-glycero-3-phosphocholine accounted for 62% of the decrease of arachidonate from choline-linked phosphoglycerides. In contrast, 60% of the release of labeled arachidonate from PC subclasses originated from 1-acyl molecular species. 1-Alk-1'-enyl-2-acyl-sn-glycero-3-PE contained 71% of the arachidonate in ethanolamine-linked phosphoglycerides and was the major PE subclass which was degraded during neutrophil activation with ionophore A23187. These findings demonstrate that human neutrophils contain large ether-linked stores of arachidonate and the capacity to mobilize these stores. In addition, this study points out major discrepancies between using mass or label to determine sources of arachidonate for eicosanoids.

Evidence that increasing the cellular content of eicosapentaenoic acid does not reduce the biosynthesis of platelet-activating factor.

Our study has examined platelet-activating factor (PAF) biosynthesis in neutrophils from individuals on a fish oil-enriched diet and in mast cells enriched with eicosapentaenoic acid (EPA) in vitro. Neutrophils isolated from males who were fed fish oil supplement (EPA; 2.8 g/day) for 5 wk contained large quantities of eicosapentaenoate in phosphatidylcholine (PC) and phosphatidylethanolamine and less in phosphatidylinositol. The ratio arachidonate/eicosapentaenoate in PC and phosphatidylethanolamine decreased from greater than 10 before the enriched diet to approximately 3 after the diet. The putative precursor of PAF, 1-O-alkyl-2-acyl-sn-glycero-3-phosphocholine (1-O-alkyl-2-acyl-GPC) contained the bulk of eicosapentaenoate in PC subclasses with smaller quantities found in 1-acyl and 1-alk-1'-enyl linked species. Ionophore A23187-stimulated neutrophils produced similar quantities of PAF before and after enriched diet. Neutrophils during normal diet acylated 1-O-alkyl-2-lyso-GPC only with arachidonate whereas neutrophils from individuals on enriched diet transferred both arachidonate and eicosapentaenoate into exogenously-provided 1-O-alkyl-2-lyso-GPC. This allowed for the labeling of neutrophils with 1-O-[3H]-alkyl-2-arachidonoyl-GPC (before diet) as well as neutrophils with 1-O-[3H]-alkyl-2-eicosapentaenoyl-GPC and 1-O-[3H]-alkyl-2-arachidonoyl-GPC (after diet). Neutrophils after diet converted similar quantities of these labeled precursors to labeled PAF upon stimulation as those before the diet. Analysis of the nature of the long chain acyl residue remaining in the sn-2 position of 1-alkyl-2-acyl-GPC after cell stimulation indicated that arachidonate and eicosapentaenoate were both released from 1-O-alkyl-2-acyl-GPC at comparable rates. Finally, in vitro supplementation of murine mast cells (PT-18) with arachidonic acid or EPA caused a marked increase in the amount of PAF produced by the cell without having any effect on histamine release. Data from these experiments suggest that EPA is incorporated into a PAF precursor pool. However, this appears not to inhibit PAF production because phospholipase A2 can use eicosapentaenoate- as well as arachidonate-containing phospholipids in the initial step of PAF biosynthesis.

Test strip meconium screening for cystic fibrosis.

The Boehringer-Mannheim Corporation (BMC) strip test is extremely reliable in indicating an albumin content above 20 mg/gm of dried meconium. All infants born during one year in 14 Milwaukee area hospitals were tested. Of 16,224 newborns, two were diagnosed correctly as suffering from cystic fibrosis and two were missed. False-positive tests were obtained in 0.9% of infants (prematurity, melena, gastroschisis, and intrauterine infection). The strip test is, at present, the best available but not the perfect screening method for cystic fibrosis.

Upper abdominal abscess: a continuing and deadly problem.

Subphrenic and other upper abdominal abscesses continue to be associated with high mortality, even in today's era of broad spectrum antibiotics and sophisticated surgical techniques. Most cases represent complications of intraabdominal surgery. Because the clinical presentation is often subtle and nonspecific, the radiologist plays a paramount role in early diagnosis. Conventional radiography remains an effective method in the initial detection of upper abdominal abscesses. In this study, radiographs were reviewed in 82 patients. In retrospect, plain films revealed extraluminal gas or soft-tissue mass due to abscess in 58 patients (71%). Of these, the abscess was accurately reported initially in 42 patients, but it was initially missed in 16 patients. Conventional gastrointestinal contrast studies, which were underutilized, proved extremely accurate in demonstrating abnormality, especially in the left upper abdomen. Computed tomography, gallium scanning, and ultrasound were effective confirmatory procedures in many cases.