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Ten patients (eight male/two female) with advanced dilated cardiomyopathy (NYHA III/IV) and a mean fractional shortening in two-dimensional echocardiogram of 20% (9 to 30%) and a mean sodium excretion of 21 mmol (8 to 40 mmol) per day, pretreated with digoxin, captopril and a mean frusemide-dose of 147 mg (80 to 500 mg) without an effective diuresis, were additional treated with 2.5 to 5 mg oral metolazone daily. All patients had a brisk diuresis within 24 hours and a mean weight loss of 8.9 kg (3 to 20 kg) until discharge. All patients improved considerably by additional metolazone-therapy. Seven patients developed a mild hyponatraemia (122 to 132 mmol/l), seven showed mild (greater than or equal to 3.2 mmol/l) and one had a serious hypokalaemia (2.8 mmol/l); spironolactone-pretreated patients developed no hypokalaemia. Notably none of the patients had serious blood pressure fluctuation or a deterioration of renal function. To avoid severe electrolyte-disturbances, additional metolazone-therapy should be practised in hospital, preferring low-dose metolazone and reducing frusemide-dose under careful biochemical monitoring after diuresis is started.

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Resistance to normal human serum (NHS) killing in Neisseria gonorrhoeae has been associated with particular types of Protein I (PI) and lipopolysaccharide (LPS), but many exceptions exist, and the role of these structures in determining serum reactivities remains controversial. In reality, the response of the gonococcus to NHS is probably governed by several parameters involving a number of outer-membrane (OM) components. We surveyed the serum reactivities of 14 strains of N. gonorrhoeae and characterized each of their major OM components. The strains presented a spectrum of sensitivity to pooled NHS. As assessed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis, immunoblotting, and peptide mapping, the strains were also quite heterogeneous in terms of PI, H.8 antigen, and LPS type, and the presence of the 2-1-L8 epitope. Five of the strains had identical PIAs in varying LPS and H.8 backgrounds, and four had identical PIBs in varying LPS and H.8 backgrounds. As assessed by electrophoretic migration and monoclonal antibody binding, Protein III and the 44,000 Dalton protein were identical in these strains. We found no association between PI subclass and serum sensitivity, while H.8 and LPS variation appeared to be related to bactericidal responses. The diversity and close interaction of gonococcal components in the OM are undoubtedly involved in differential abilities to survive NHS killing.

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Pooled normal human serum killing of 14 strains of Neisseria gonorrhoeae was assessed by dilution plate and microtiter methods. In both assays, the strains presented a spectrum of sensitivity to the serum. In the dilution plate assay, results with two different concentrations of human serum were similar for most, but not all of the strains tested. When data for all of the strains were compared, no correlation was found between the dilution plate and microtiter bactericidal assays. Finally, we found that the bactericidal capacities of intact and complement-depleted human sera were very similar when assessed by microtiter methods, suggesting a non-complement-mediated serum killing mechanism.

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The composition and patterns of metabolism of phospholipids isolated as part of a lipid-depleted membrane fragment (LDM fragment) and associated with the membrane adenosine triphosphatase complex have been compared with those of the bulk membrane phospholipid. The bulk lipid was extracted from washed membranes with sodium cholate. The LDM fragments, which contained a portion of the electron transport system and the membrane adenosine triphosphatase complex, were purified by chromatography with Sepharose 6B. The LDM fragment preparations contained 0.10 +/- 0.02 mumol of lipid phosphorus per mg of protein, compared with 0.54 +/- 0.05 mumol of lipid phosphorus per mg of protein for washed membranes. The phospholipid associated with the LDM fragments consisted of 78 +/- 4% cardiolipin, 7 +/- 1% phosphatidylglycerol, and 15 +/- 3% phosphatidylethanolamine. Changes in the total membrane lipid composition (produced by culture conditions) did not alter the phospholipid composition of the LDM fragments. The adenosine triphosphate complex was separated from the other components of the LDM fragments by suspension of the fragments in 1% Triton X-100 and precipitation with antibody specific for the F(1) component of the adenosine triphosphatase complex. The phospholipid isolated with the adenosine triphosphatase complex consisted of 86% cardiolipin, 8% phosphatidylglycerol, and 6% phosphatidylethanolamine. In pulse-chase experiments with (32)P and [2-(3)H]glycerol, the labeling patterns of the phosphatididylglycerol and phosphatidylethanolamine associated with the LDM fragments were different from those of the bulk membrane phosphatidylglycerol and phosphatidylethanolamine. It was concluded that at least a portion of the phospholipid isolated with the LDM fragments was part of a native lipid-protein complex.

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