Differences in contributing factors to tuberculosis incidence in U.S. -born and foreign-born persons.

To determine the factors contributing to tuberculosis incidence in the U.S.-born and foreign-born populations in San Francisco, California, and to assess the effectiveness of tuberculosis control efforts in these populations, we performed a population-based molecular epidemiologic study using 367 patients with strains of Mycobacterium tuberculosis recently introduced into the city. IS6110-based and PGRS-based restriction fragment length polymorphism (RFLP) analyses were performed on M. tuberculosis isolates. Patients whose isolates had identical RFLP patterns were considered a cluster. Review of public health and medical records, plus patient interviews, were used to determine the likelihood of transmission between clustered patients. None of the 252 foreign-born cases was recently infected (within 2 yr) in the city. Nineteen (17%) of 115 U. S.-born cases occurred after recent infection in the city; only two were infected by a foreign-born patient. Disease from recent infection in the city involved either a source or a secondary case with human immunodeficiency virus (HIV) infection, homelessness, or drug abuse. Failure to identify contacts accounted for the majority of secondary cases. In San Francisco, disease from recent transmission of M. tuberculosis has been virtually eliminated from the foreign-born but not from the U.S.-born population. An intensification of contact tracing and screening activities among HIV-infected, homeless, and drug-abusing persons is needed to further control tuberculosis in the U.S.-born population. Elimination of tuberculosis in both the foreign-born and the U.S. -born populations will require widespread use of preventive therapy.

Invasive Haemophilus influenzae infections in men with HIV infection.

OBJECTIVE: To determine the incidence of invasive Haemophilus influenzae disease in men with the acquired immunodeficiency syndrome (AIDS) or human immunodeficiency virus (HIV) infection and the proportion of disease due to serotype b. DESIGN: Population-based, active surveillance. SETTING: San Francisco (Calif) Department of Health. PARTICIPANTS: All men 20 to 49 years of age with invasive H influenzae disease. RESULTS: The cumulative incidences of invasive H influenzae disease in men 20 to 49 years of age with AIDS and in HIV-infected men 20 to 49 years of age without AIDS were 79.2 and 14.6 per 100,000, respectively, but only 33% of cases were due to serotype b. The corresponding rates for invasive H influenzae b disease were 11.3 and 7.6 per 100,000. CONCLUSIONS: Men with AIDS or HIV infection are at increased risk of invasive H influenzae infections, including H influenzae b, but such infections are still infrequent in this population.

Ripple effects of PPS on nursing homes: swimming or drowning in the funding stream?

In a sample of 189 nursing homes, 83% reported that Medicare's hospital Prospective Payment System (PPS) impacts patient needs, 53% said it impacts both patients and services provided, and 25% said it impacts referrals to hospitals. The PPS effects were found to depend on the facility factors of size, Medicare certification, and tax status, and on the local market area factors of prevalence of PPS-nonexcluded hospital psychiatric units, competition, and levels of hospital discharges.

Macro creatine kinases: results of isoenzyme electrophoresis and differentiation of the immunoglobulin-bound type by radioassay.

In 2.9% of sera from 1253 unselected patients we detected two different types of macromolecular creatine kinases (CK; EC 2.7.3.2). One macro type was represented by immunoglobulin-linked CK: in sera containing macro CK-BB isoenzyme, 125I-labeled CK-BB was bound with high affinity to the immunoglobulin fraction. Furthermore, during electrophoresis, macro CK-BB mostly migrated between CK-MB and CK-MM, and was fixed to Protein A from Staphylococcus aureus. We therefore propose radioelectrophoresis as a specific, highly sensitive, and simple method for detecting this type of macro CK. This form occurs predominantly in elderly women, is not correlated to any specific disease, and persists in blood over a long period of time. In contrast, a second type (macro-CK type 2) never bound radiolabeled CK isoenzymes, and was not adsorbed to Protein A. Electrophoretic migration of this macro-CK type 2 was generally cathodic to CK-MM. We observed this type in severely ill patients, frequently those suffering from malignant tumors. Clinical observations and biochemical data suggest that macro-CK type 2 is of mitochondrial origin.

Macro creatine kinase: determination and differentiation of two types by their activation energies.

Determination of the MB isoenzyme of creatine kinase in patients with acute myocardial infarction may be disturbed by the presence of macro creatine kinase. The relative molecular mass of this form of creatine kinase in human serum is at least threefold that of the ordinary enzyme, and it is more thermostable. Here we describe our method for determination of macro creatine kinases and an easy-to-perform test for differentiating two forms of macro creatine kinase, based on their distinct activation energies. The activation energies of serum enzymes are mostly in the range of 40-65 kJ/mol of substrate. Unlike normal cytoplasmatic creatine kinases and IgG-linked CK-BB (macro creatine kinase type 1) a second form of macro creatine kinase (macro creatine kinase type 2) shows activation energies greater than 80 kJ/mol of substrate. The exact composition of macro creatine kinase type 2 is still unknown, but there is good reason to believe that it is of mitochondrial origin.

Stability of macro creatine kinases and creatine kinase isoenzymes compared: heat inactivation test for determination of thermostable creatine kinases.

Creatine kinase isoenzymes showed decreasing thermal stability and increasing lability towards pH changes in the order: MM, MB, and BB. The three isoenzymes exhibited their highest stability between pH 6.5 and 7.0. At 37 degrees C and an almost physiological pH of 7.5 the decay constants were 0.025, 0.164 and 0.580 h-1 (MM, MB, and BB isoenzyme), respectively. In contrast to free creatine kinase BB, immunoglobulin-linked creatine kinase BB (macro creatine kinase BB, type 1 macro creatine kinase) showed a markedly higher stability; this accounts for the persistence of creatine kinase BB activity in macro creatine kinasaemia. In addition we identified a second type of macro creatine kinase in patients' sera, which is also thermostable. A simple heat inactivation test (20 minutes, 45 degrees C, immunoinhibition of the M-subunits) differentiates thermostable macro creatine kinases from thermolabile creatine kinases and thus completes isoenzyme diagnosis.

Macro creatine kinase BB: evidence for specific binding between creatine kinase BB and immunoglobulin G.

In the sera of four female patients with proven IgG-linked creatine kinase BB (macro creatine kinase BB) we studied the nature of the binding between creatine kinase isoenzymes and immunoglobulin G. The specificity towards the isoenzyme bound, the recombination of the complex after treatment with acid buffer, the site of the binding between both partners and an apparently monoclonal nature of the immunoglobulin G involved in the complex indicate that a specific antigen-antibody reaction is responsible for the existence of macro creatine kinase BB in the sera investigated.

Mechanism of phagocytosis in Dictyostelium discoideum: phagocytosis is mediated by different recognition sites as disclosed by mutants with altered phagocytotic properties.

The recognition step in the phagocytotic process of the unicellular amoeba dictyostelium discoideum was examined by analysis of mutants defective in phagocytosis, Reliable and simple assays were developed to measure endocytotic uptake. For pinocytosis, FITC-dextran was found to be a suitable fluid-phase marker; FITC-bacteria, latex beads, and erythrocytes were used as phagocytotic substrates. Ingested material was isolated in one step by centrifuging through highly viscous poly(ethyleneglycol) solutions and was analyzed optically. A selection procedure for isolating mutants defective in phagocytosis was devised using tungsten beads as particulate prey. Nonphagocytosing cells were isolated on the basis of their lower density. Three mutant strains were found exhibiting a clear-cut phenotype directly related to the phagocytotic event. In contrast to the situation in wild-type cells, uptake of E. coli B/r by mutant cells is specifically and competitively inhibited by glucose. Mutant amoeba phagocytose latex beads normally but not protein-coated latex, nonglucosylated bacteria, or erythrocytes. Cohesive properties of mutant cells are altered: they do not form EDTA-sensitive aggregates, and adhesiveness to glass or plastic surfaces is greatly reduced. Based upon these findings, a model for recognition in phagocytosis is proposed: (a) A lectin-type receptor specifically mediates binding of particles containing terminal glucose (E. coli B/r). (b) A second class of "nonspecific" receptors mediate binding of a variety of particles by hydrophobic interaction. Nonspecific binding is affected by mutation in such a way that only strongly hydrophobic (latex) but not more hydrophilic particles (e.g., protein-coated latex, bacteria, erythrocytes) can be phagocytosed by mutant amoebae.

Complementation in vitro of mutant and wild-type ATPase of Escherichia coli using isolated subunits.

1. The inactive ATPases of four different mutant strains of Escherichia coli have been purified to homogeneity. 2. Molecular weights, subunit patterns in sodium dodecylsulfate electrophoresis and immunological properties of mutant and wild-type proteins are identical. The mutant enzymes compete with the wild-type enzyme for the binding sites on the membrane. 3. On freezing and thawing in salt solutions, the ATPase is split into subunits IA (alpha, gamma, epsilon), IB (delta; alpha, gamma, epsilon), and II (beta). By complementation in vitro of the isolated subunits, it is shown that subcomplex IA (alpha, gamma, epsilon) is altered in the mutant strains described here.

ATPase of Escherichia coli: purification, dissociation, and reconstitution of the active complex from the isolated subunits.

A simple procedure for the purification of Mg2+-stimulated ATPase of Escherichia coli by fractionation with poly(ethylene glycols) and gel filtration is described. The enzyme restores ATPase-linked reactions to membrane preparations lacking these activities. Five different polypeptides (alpha, beta, gamma, delta, epsilon) are observed in sodium dodecyl sulfate electrophoresis. Freezing in salt solutions splits the enzyme complex into subunits which do not possess any catalytic activity. The presence of different subunits is confirmed by electrophoretic and immunological methods. The active enzyme complex can be reconstituted by decreasing the ionic strength in the dissociated sample. Temperature, pH, protein concentration, and the presence of substrate are each important determinants of the rate and extent of reconstitution. The dissociated enzyme has been separated by ion-exchange chromatography into two major fragments. Fragment IA has a molecular weight of about 100000 and contains the alpha, gamma, and epsilon polypeptides. The minor fragment, IB, has about the same molecular weight but contains, besides alpha, gamma, and epsilon, the delta polypeptide. Fragment II, with a molecular weight of about 52000, appears to be identical with the beta polypeptide. ATPase activity can be reconstituted from fragments IA and II, whereas the capacity of the ATPase to drive energy-dependent processes in depleted membrane vesicles is only restored after incubation of these two fractions with fraction IB, which contains the delta subunit.