Mechanical treatment of plantar fasciitis. A prospective study.

A randomized, prospective study was conducted to compare the effectiveness of three individual mechanical modalities in the treatment of plantar fasciitis. Two hundred fifty-five subjects were randomly assigned to one of three treatment groups: custom-made orthoses, over-the-counter arch supports, or tension night splints. Subjects were treated for 3 months, with follow-up visits at 2, 6, and 12 weeks. No statistically significant difference was noted among treatment groups with respect to final outcomes based on first-step pain or pain felt during the day. However, there was a statistically significant difference among the three groups with respect to early patient withdrawal from the study due to continued severe pain, noncompliance, or inability to tolerate the device. Patient compliance was greatest with the use of custom-made orthoses.

Clinical evaluation comparing AxSYM CA 15-3, IMx CA 15-3 and Truquant BRTM RIA.

A retrospective clinical study was conducted to compare results obtained by AxSYM(R) CA 15-3(TM), IMx(R) CA 15-3 and Truquant(R) BRTM RIA using surplus serum specimens from healthy volunteers and patients with benign and malignant diseases. Linear regression analysis of AxSYM and IMx CA 15-3 versus Truquant BR RIA for specimens with results 0-250 U/ml gave correlation coefficients of 0. 888 and 0.910 and slopes of 0.67 and 0.69, respectively. For specimens with results 0-2,000 U/ml, slopes were 0.95 and 0.91, respectively. Receiver operator characteristic analyses, based on results from healthy females plus nonmalignant disease patients versus breast cancer patients, for all three assays gave essentially equivalent areas under the curves. Concordance between AxSYM or IMx CA 15-3 and Truquant BR RIA was greater than 92%. Serial dilution of seven serum specimens yielded linear regression correlation coefficients ranging from 0.997 to 1.000 for AxSYM and IMx CA 15-3, and from 0.962 to 0.998 for Truquant BR RIA. The average percent CVs of the calculated assay values for the 7 specimens were 4.9, 2.7 and 18.1 for AxSYM CA 15-3, IMx CA 15-3 and Truquant BR RIA, respectively. Average percent recoveries ranged from 96.2 to 110.7 for AxSYM and IMx CA 15-3, and 81.8 to 93.3 for Truquant BR RIA. Although assay values differ between the two methodologies, AxSYM CA 15-3, IMx CA 15-3 and Truquant BR RIA showed comparable trending results for the 24 breast cancer patients evaluated for serial monitoring.

Conservative treatment of plantar fasciitis. A prospective study.

A randomized, prospective study was conducted to compare the individual effectiveness of three types of conservative therapy in the treatment of plantar fasciitis. One hundred three subjects were randomly assigned to one of three treatment categories: anti-inflammatory, accommodative, or mechanical. Subjects were treated for 3 months, with follow-up visits at 2, 4, 6, and 12 weeks. For the 85 patients who completed the study, a statistically significant difference was noted between groups, with mechanical treatment with taping and orthoses proving to be more effective than either anti-inflammatory or accommodative modalities.

Acetylcholine-induced and nitric oxide-mediated vasodilation in burns.

Microvascular endothelial cells actively participate in local regulation of blood flow and blood-tissue exchange by producing various vasoactive substances including nitric oxide (NO). This study examined microcirculatory changes in the early stage of thermal injury and the NO-related mechanisms. Resistance arterioles of rat cremaster muscle were observed using intravital microscopy. Arteriolar diameter and flow velocity were measured and flow rate was calculated after administration of various vasoactive agonists in burns. In fluid-resuscitated rats with stable systemic blood pressure, microvascular caliber and blood flow were not significantly altered in the first hour following a 25% total body surface area full-thickness scald burn. Topical application of acetylcholine (ACh), an endothelium-dependent vasodilator, increased arteriolar diameter and flow rate in a dose-dependent fashion. The dose-responsive effects of ACh were significantly greater in burned rats than in sham-burned rats, and the augmentation was blocked by inhibition of NO production with NG-monomethyl-l-arginine (L-NMMA). Topical application of adenosine, an endothelium-independent vasodilator, and sodium nitroprusside, an exogenous NO donor, markedly increased arteriolar diameter and flow rate. The effects were not significantly different in burned and sham-burned animals, and the adenosine-induced vasodilation was not blocked by L-NMMA. These data suggest that endothelium-dependent and NO-mediated arteriolar dilation is enhanced in the early stage of thermal injury. This effect may play an important role in the pathophysiological events of microcirculation and blood-tissue exchange in burns.

Inhibition of natural killer cells by a cytomegalovirus MHC class I homologue in vivo.

Herpesviruses, such as murine and human cytomegalovirus (MCMV and HCMV), can establish a persistent infection within the host and have diverse mechanisms as protection from host immune defences. Several herpesvirus genes that are homologous to host immune modulators have been identified, and are implicated in viral evasion of the host immune response. The discovery of a viral major histocompatibility complex (MHC) class I homologue, encoded by HCMV, led to speculation that it might function as an immune modulator and disrupt presentation of peptides by MHC class I to cytotoxic T cells. However, there is no evidence concerning the biological significance of this gene during viral infection. Recent analysis of the MCMV genome has also demonstrated the presence of a MHC class I homologue. Here we show that a recombinant MCMV, in which the gene encoding the class I homologue has been disrupted, has severely restricted replication during the acute stage of infection compared with wild-type MCMV. We demonstrate by in vivo depletion studies that natural killer (NK) cells are responsible for the attenuated phenotype of the mutant. Thus the viral MHC class I homologue contributes to immune evasion through interference with NK cell-mediated clearance.

Identification and characterization of a G protein-coupled receptor homolog encoded by murine cytomegalovirus.

This report describes the identification of a murine cytomegalovirus (MCMV) G protein-coupled receptor (GCR) homolog. This open reading frame (M33) is most closely related to, and collinear with, human cytomegalovirus UL33, and homologs are also present in human herpesvirus 6 and 7 (U12 for both viruses). Conserved counterparts in the sequenced alpha- or gammaherpesviruses have not been identified to date, suggesting that these genes encode proteins which are important for the biological characteristics of betaherpesviruses. We have detected transcripts for both UL33 and M33 as early as 3 or 4 h postinfection, and these reappear at late times. In addition, we have identified N-terminal splicing for both the UL33 and M33 RNA transcripts. For both open reading frames, splicing results in the introduction of amino acids which are highly conserved among known GCRs. To characterise the function of the M33 in the natural host, two independent MCMV recombinant viruses were prepared, each of which possesses an M33 open reading frame which has been disrupted with the beta-galactosidase gene. While the recombinant M33 null viruses showed no phenotypic differences in replication from wild-type MCMV in primary mouse embryo fibroblasts in vitro, they showed severely restricted growth in the salivary glands of infected mice. These data suggest that M33 plays an important role in vivo, in particular in the dissemination to or replication in the salivary gland, and provide the first evidence for the function of a viral GCR homolog in vivo.

Reinventing government for the clinical lab.

Learn how the power of teamwork fueled by a common goal enabled members of a California government agency to educate thousands of diverse clinical laboratory scientists--despite unanticipated dilemmas and limited resources.

Studies on the polymorphism of the fifth component of complement in laboratory mice.

The fifth component of complement C5, as well as C3, plays an important role in the inflammatory response. Since our previous studies have implicated polymorphism of C3 in disease pathogenesis, similar studies were undertaken on C5 using inbred and outbred laboratory mice. Consistent with previous findings, male mice were shown to have higher serum C5 concentrations than female mice. In male SJL/J mice, however, the serum concentration of C5 was significantly greater than in males of other strains. An immunofixation electrophoretic method was therefore developed to determine whether SJL/J mice had a different electrophoretic form of C5. The electrophoretic mobility of C5 in serum from SJL/J mice was similar to that in other strains. Some mice, including NOD/Lt, Quackenbush and ARC, had complete C5 deficiency. Southern blotting showed that C5 deficiency in these mice was associated with the same gene rearrangement found in other C5-deficient laboratory mice. No difference in the structure of the C5 gene was seen between SJL/J mice and other C5-sufficient inbred strains. In contrast, C5-sufficient Quackenbush and ARC outbred mice had a unique C5 RFLP pattern.

Studies on the structure of complement C3 and the stability of C3 derived phagocytic ligands C3b/iC3b in SJL/J and BALB/c mice.

Female SJL/J mice are more susceptible to development of experimental autoimmune myositis than most other mouse strains. Since complement has been implicated in the pathogenesis of inflammatory muscle disease in humans, quantitative and qualitative studies of complement C3 were undertaken in SJL/J and BALB/c mice to determine whether complement may influence disease susceptibility in SJL/J mice. In accordance with previous studies, mature male and female BALB/c mice were shown to have similar serum C3 concentrations. However, differences were found between mature male and female SJL/J mice. Male SJL/J mice have significantly higher serum C3 concentrations than SJL/J females and both sexes of BALB/c mice suggesting that serum C3 concentration may be variably influenced by sex in some mouse strains. Qualitatively, SJL/J mice were shown to have a different allotypic form of C3 (C3F) compared to the common electrophoretically slow form (C3S) found in BALB/c mice and most other mouse strains. Furthermore, studies on the decay rate of C3 revealed that C3b/iC3b fragments are converted to C3c/d at a faster rate in sera from female SJL/J mice compared to female BALB/c mice. Because removal and solubility of immune complexes is influenced by complement C3, it is possible that the more rapid decay of the phagocytic ligands C3b/iC3b may account for the increased susceptibility to development of autoimmune disease in female SJL/J mice.

Genetic, structural, and functional studies of a C3-like protein in the marsupial Setonix brachyurus.

A monospecific goat antiserum was prepared against a putative C3 protein (QuC3) from serum of the Western Australian macropod Setonix brachyurus (quokka) using the classical method previously used to produce antiserum against C3 from other mammalian species. Sodium dodecyl sulphate-polyacrylamide gel electrophoretic analysis of reduced immunoprecipitated QuC3 revealed two polypeptide chains with an estimated M(r) of 128,000 and 82,000, respectively--presumably reflecting subunit molecules similar to the C3 alpha and beta subunits found in other mammalian species. No variation in the size of either of the QuC3 alpha or beta subunits was found in different quokka serum samples. Furthermore, no variation in the electrophoretic mobility of QuC3 was detected amongst the same samples using a standard immunofixation electrophoretic technique. The addition of zymosan or immune complexes to fresh quokka serum resulted in activation of QuC3. Immune complex mediated activation of QuC3 was inhibited by the addition of EDTA and Mg(2+)-EGTA. By contrast, EDTA only inhibited activation of QuC3 following incubation of fresh quokka serum with zymosan, suggesting that the quokka has a classical and an alternative C3 activation pathway, similar to those found in eutherian mammals. This study has shown for the first time that one of the macropod marsupials (S. brachyurus) has a C3-like protein with similar structural and functional characteristics to C3 found in eutherian mammals. However, unlike C3 in other mammalian species, genetic variability of the structure of the quokka C3-like protein may be restricted.

Antisperm antibody binding to human acrosin: a study of patients with unexplained infertility.

OBJECTIVE: Antisperm antibody binding to acrosin was investigated by Western Blotting. The clinical significance of this binding specificity was assessed in a 2-year clinical follow-up. DESIGN: Consecutive serum samples positive for antisperm antibodies by both enzyme-linked immunosorbent assay and immunobead testing were evaluated for acrosin-binding specificity. SETTING: The patients were followed in an outpatient setting by private infertility specialists. PATIENTS: Sixty-five consecutive infertile referral patients with positive antisperm antibody were evaluated. Clinical follow-up was obtained on 8 of 9 females with evidence of antibody binding to acrosin and 19 of 26 females with no specific binding to acrosin. INTERVENTIONS: Prednisone therapy was given during six courses of intrauterine insemination with husband's sperm. All treatment decisions were made by private physicians independent of the acrosin-binding result. MAIN OUTCOME MEASURES: Pregnancy status was obtained as part of a 2-year follow-up. RESULTS: Acrosin-binding specificity was demonstrated in 10 (15%) of the 65 patients. Two of the 8 women (25%) with antibody binding to acrosin and 6 of the 19 women (32%) with antisperm antibodies but no specific binding to acrosin delivered normal children. CONCLUSIONS: Although antibody-binding specificity to acrosin could be demonstrated, a 2-year clinical follow-up showed no difference in pregnancy rates when compared with women with antisperm antibodies showing no binding specificity to acrosin.

Murine candidiasis: sex differences in the severity of tissue lesions are not associated with levels of serum C3 and C5.

Mice deficient in the fifth component of complement are known to be extremely susceptible to lethal challenge with Candida albicans. However, male mice, that have significantly higher concentrations of serum C5 than females, were markedly more susceptible to infection. This difference was observed in both susceptible (CBA/H) and resistant (BALB/c) mice. Levels of serum C3 likewise showed no correlation with susceptibility.

An enzyme immunoassay for carcinoembryonic antigen which employs simultaneous incubation of specimen with solid phase and enzyme-conjugated antibodies.

The Abbott CEA-EIA Monoclonal One-Step procedure was evaluated and compared to the Abbott CEA-EIA Monoclonal, a two-step assay. The reproducibility, sensitivity, recovery and dilution linearity were similar for both assays. Excellent correlations were found between the carcinoembryonic antigen (CEA) values obtained with these assays for healthy donors (n = 261, r = 0.951), patients with benign diseases (n = 171, r = 0.994) and cancer patients (n = 585, r = 0.997). In serial monitoring studies, one-step CEA values paralleled the CEA values determined with the two-step assay regardless of the type of cancer. The advantages of the one-step CEA assay over the two-step assay include significant reduction in both attended and total assay times, reduced sample volume, dry antibody-coated beads, color-coded reagents and an extended range for the standard curve. These test improvements were achieved without affecting the reproducibility, sensitivity and accuracy of the CEA measurement.

Comparison of a direct and indirect ELISA for quantitating antisperm antibody in semen.

A direct and an indirect quantitative ELISA for antisperm antibody were compared using the spermatozoa and cell-free seminal fluid of 66 infertile males. The normal concentration of sperm binding immunoglobulin was less than or equal to 1.5 fg Ig per spermatozoon for the indirect seminal plasma assay and less than or equal to 1.5 fg Ig per spermatozoon by the direct assay. Of the 66 infertile males, 21% (14/66) had elevated levels of antisperm antibody in their seminal plasma and 26% (17/66) had elevated levels bound directly to their spermatozoa. The direct correlation between the results of these assays was 94%. A simple linear regression analysis between the indirect and direct measurements of antisperm antibody resulted in a correlation coefficient of r = 0.907. There was no statistically significant difference between results from the direct and indirect methods of the patients as a group. However, there was evidence of autospecificity in a small percentage of males who had elevated levels of antisperm antibody by the direct assay that was not detected by the indirect assay using pooled donor spermatozoa.

Platelet antibody binding in systemic lupus erythematosus.

Using a combination of enzyme-linked immunosorbent assay and SDS-PAGE with protein blot (Western blotting), increased levels of serum platelet bindable immunoglobulin (SPBIg) were demonstrated in 10 of 10 thrombocytopenic patients with systemic lupus erythematosus (SLE) (7.0-60 fg Ig/platelet) with consistent binding to SDS-PAGE platelet fractions of approximate molecular weight (120 and 80 kDa). This pattern of Ig binding was characteristic of SLE and was not seen in 20 normal volunteers and infrequently seen in 20 patients with idiopathic thrombocytopenic purpura.