Biochemical analysis of human milk treated with sodium dodecyl sulfate, an alkyl sulfate microbicide that inactivates human immunodeficiency virus type 1.

Reduction of transmission of human immunodeficiency virus type 1 (HIV-1) through human milk is needed. Alkyl sulfates such as sodium dodecyl sulfate (SDS) are microbicidal against HIV-1 at low concentrations, have little to no toxicity, and are inexpensive. The authors have reported that treatment of HIV-1-infected human milk with < or = 1% (10 mg/mL) SDS for 10 minutes inactivates cell-free and cell-associated virus. The SDS can be removed with a commercially available resin after treatment without recovery of viral infectivity. In this article, the authors report results of selective biochemical analyses (ie, protein, immunoglobulins, lipids, cells, and electrolytes) of human milk subjected to SDS treatment and removal. The SDS treatment or removal had no significant effects on the milk components studied. Therefore, the use of alkyl sulfate microbicides to treat milk from HIV-1-positive women may be a simple, practical, and nutritionally sound way to prevent or reduce transmission of HIV-1 while still feeding with mother's own milk.     

Candida albicans growth studies: a hypothesis for the pathogenesis of Candida infections in burns.

The proteolytic environment in which Candida albicans exists strongly affects its virulence. To determine whether virulence might be related to C. albicans growth in different proteolytic environments, we measured renal fungal load in burned mice and found significantly greater Candida census in kidneys from mice that were challenged with a high proteinase-generating parent C. albicans (MY 1044) versus those that were challenged with its low proteinase-generating mutant (MY 1049). In vitro, MY 1044 cells grew faster than MY 1049 cells in media that contained sera from burned mice as the only nitrogen source. Augmentation of media with proteinase or a mixture of amino acids increased growth of MY 1049 cells, whereas augmentation with proteinase inhibitor decreased MY 1044 growth. In conclusion, in vitro growth of both the mutant and its parent strain was affected by the proteolytic environment in which they existed; thus, virulence differences for MY 1044 and MY 1049 could be due in part to growth differences between these two strains in different proteolytic environments. These results were combined with existing observations, and we proposed a theory for the pathogenesis of C. albicans in burns.     

Prevalence of juvenile periodontitis in a circumpubertal population

A cross-sectional radiographic screening was performed on bite-wing pairs (BW) from 1872 10-12 year old schoolchildren in the Greater Worcester, Massachusetts area to assess the prevalence of juvenile periodontitis (JP). The 3-stage screening process entailed: (1) visual identification of possible cases based upon a visual assessment of BW for interproximal crestal bone levels greater than or equal to 2 mm from the cemento-enamel junction (CEJ) on greater than or equal to 1 permanent first molar; (2) identification of probable cases based upon BW from possible cases measured with a transparent ruler calibrated in millimeters; (3) finally, clinical confirmation of JP in consenting probable cases. A total of 1038 subjects were eligible to be included in the study (greater than or equal to 3 mesial sites readable). Of the 1038 eligible subjects, 117 possible and 103 probable cases were identified in stage 1 and stage 2, respectively. A total of 99 probable cases could be contacted and 43 were examined clinically. Two cases of JP were confirmed clinically in stage 3, yielding a prevalence rate of 4.6/1000. Specifically, this report defines a rate of JP in 10-12 year-old schoolchildren for the first time. In addition, these results indicate that BW can be used to identify children with JP from large data sets. However, further studies including complete clinical and radiographic examinations are necessary to determine whether this method is adequate for large epidemiologic studies.     

Immunoassays for quantifying choriogonadotropin compared for assay performance and clinical application.

We examined calibration and accuracy, precision, sensitivity, specificity, and "hook" effects for recently revised automated choriogonadotropin (hCG) immunoassay systems (Baxter-Dade Stratus II, Abbott IMx intact hCG and total beta hCG) and compared them with a widely used immunoradiometric assay (Hybritech). We estimated hCG in pregnant women, women with trophoblastic disease, nonpregnant young and menopausal women, normal men, and men with testicular tumors. We found clinically unimportant differences in calibration (all calibrated to the 3rd International Standard). Detection of hCG by all four assays was limited by their responses in serum from nonpregnant women and men. Precision within-run was best for the automated instruments, but all four assays had similar between-run precision. The Hybritech, Stratus, and IMx intact assays are specific for intact hCG. The IMx total beta assay quantifies both free beta subunit and beta subunit present in intact hCG. There is a clinically important hook effect in the Hybritech assay but not the Stratus or IMx assays (to 1.2 x 10(6) int. units/L). Results for pregnant women were similar by all four assays. We measured "hCG" to 8 int. units/L in menopausal women, which weakly correlated with concentrations of lutropin and follitropin and was, in part, explained by crossreactivity. There was no sample-probe carryover in either instrument. We found the IMx diluting module as well as results at the extremes of the IMx calibration curves (less than 10, 800-1200 int. units/L) unreliable but encountered no such problems with the Stratus system. Both automated systems involve batch analyzers with limited throughput but provide hCG concentration estimates much more quickly than the Hybritech assay can.     

Combined reflectance and fluorescence spectroscopy for in vivo detection of cervical pre-cancer

Optical technologies, such as reflectance and fluorescence spectroscopy, have shown the potential to provide improved point-of-care detection methods for cervical neoplasia that are sensitive, specific, and cost-effective. Our specific goals are to analyze the diagnostic potential of reflectance and fluorescence spectra, alone and in combination, to discriminate normal and precancerous cervical tissue in vivo and to identify which classification features contain significant diagnostic information. Reflectance spectra are measured at four source-detector separations and fluorescence emission spectra are measured at 16 excitation wavelengths, from 324 sites in 161 patients. These 20 spectral features are permuted in all possible combinations of one, two, and three; and classification algorithms are developed to evaluate the diagnostic performance of each combination. Algorithms based on fluorescence spectra alone yield better diagnostic performance than those based on reflectance spectra alone. The combination of fluorescence and reflectance do not significantly improve diagnostic performance compared to fluorescence alone, except in the case of discriminating high-grade precancers from columnar normal tissue. In general, fluorescence emission spectra at 330- to 360-nm and 460- to 470-nm excitation provide the best diagnostic performance for separating all pairs of tissue categories.     

Gene conversion is a likely cause of mutation in PKD1

Approximately 70% of the gene responsible for the most common form of autosomal dominant polycystic kidney disease ( PKD1 ) is replicated in several highly homologous copies located more proximally on chromosome 16. We recently have described a novel technique for mutation detection in the duplicated region of PKD1 that circumvents the difficulties posed by these homologs. We have used this method to identify two patients with a nearly identical cluster of base pair substitutions in exon 23. Since pseudogenes are known to be reservoirs for mutation via gene conversion events for a number of other diseases, we decided to test whether these sequence differences in PKD1 could have arisen as a result of this mechanism. Using changes in restriction digest patterns, we were able to show that these sequence substitutions are also present in N23HA, a rodent-human somatic cell hybrid that contains only the PKD1 homologs. Moreover, these changes were also detected in total DNA from several affected and unaffected individuals that did not harbor this mutation in their PKD1 gene copy. This is the first example of gene conversion in PKD1 , and our findings highlight the importance of using gene-specific reagents in defining PKD1 mutations.      

Differential effects of sympathetic activity on AV junctional automaticity and AV conduction

Summary Rapid ventricular response during episodes of supraventricular tachycardia are often followed, on abrupt cessation of the tachycardia, by prolonged pauses terminated by a sluggish and sometimes erratic escape of a supraventricular pacemaker. Such chronotropic-dromotropic paradoxes are readily reproduced in the animal laboratory following elimination of the sinus node and bilateral decentralization of the stellate ganglia and vagi. This study examined whether left stellate stimulation (0.5, 1, 2, 4, 8 and 16 Hz) or lack thereof differentially affected AV junctional automaticity and AV conduction. In the absence of any sympathetic neural activity (maximal sympathetic deficit), the AV junctional rate averaged a mere 22±2 percent of its peak performance, whereas under the same conditions, anterograde AV conduction averaged 73±5 percent and retrograde VA conduction 56 ±13 percent of their respective peak performances. On comparing the response curve (normalized responses) for AV junctional automaticity with that obtained for anterograde AV conduction the differences were significant at all frequencies between 0 and 4 Hz. Retrograde VA conduction (as assessed by the fastest ventricular pacing rate still conducted 11 to the atria) was always significantly less than anterograde AV conduction (as assessed by the fastest atrial pacing still conducted 11 to the ventricles). These results indicate that AV junctional automaticity is considerably more affected by sympathetic deficit than are either anterograde or retrograde AV conduction. In other words, AV junctional automaticity is far more dependent upon sympathetic input than AV conduction. While sympathetic influence is critical to the escape and maintenance of AV junctional automaticity both anterograde and retrograde AV conduction are remarkably resilient even under conditions of severe sympathetic deficit.