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Molecular analysis of the 18S rRNA gene of Cryptosporidium parasites from patients with or without human immunodeficiency virus infections living in Kenya, Malawi, Brazil, the United Kingdom, and Vietnam.

An 840-bp fragment of the 18S rRNA gene was used to identify Cryptosporidium spp. recovered from human immunodeficiency virus (HIV)-infected and -uninfected patients from Kenya, Malawi, Brazil, the United Kingdom, and Vietnam. Initial identification was by Ziehl-Neelsen acid-fast staining. Confirmation was by nested PCR, targeting the most polymorphic region of the 18S rRNA gene. Genotyping was by restriction endonuclease digestion of the PCR product followed by nucleotide sequencing. Among 63 isolates analyzed, four genotypes of Cryptosporidium were identified; 75% of the isolates were of the C. parvum human genotype, while the potentially zoonotic species were of the C. parvum bovine genotype (21.7%), the C. meleagridis genotype (1.6% [one isolate]), and the C. muris genotype (1.6% [one case]). HIV-infected individuals were more likely to have zoonotic genotypes than the HIV-uninfected individuals. Among the C. parvum group, strains clustered distinctly into either human or bovine genotypes regardless of the geographical origin, age, or HIV status of the patients. The intragenotypic variation observed in the C. parvum human genotype was extensive compared to that within the C. parvum bovine genotype group. The variation within genotypes was conserved in all geographical regions regardless of the patients' HIV status. The extensive diversity within genotypes at the 18S rRNA gene locus may limit its application to phylogenetic analyses.

AIDS-Related Opportunistic Infections↗

Differentiation in human myeloblastic leukemia studied in cell culture.

Normal adult hemopoiesis orginates in pluripotent stem cells; among the early differentiated descendents of such cells are progenitors committed to the erythropoietic, granulopoietic, or megakaryocytic pathways of myeloid differentiation. These may be detected in cell culture by developmental techniques, in which progenitors form colonies in viscid or semisolid media in response to appropriate stimulation. Certain diseases of hemopoiesis also originate in pluripotent stem cells; these include chronic myeloblastic leukemia, acute myeloblastic leukemia, polycythemia vera, and idiopathic myelofibrosis-the clonal hemopathies. The hypothesis is advanced that the distribution of cell classes among patients with clonal hemopathies is determined both by the differentiation potential of each pluripotent stem cell maintaining an abnormal clone and by random events occurring during clonal expansion. The latter process may account for the large variations observed between patients when committed progenitors are assayed in cultures of marrow from patients with acute myeloblastic leukemia (AML). This variation may also be used to estimate lineage relationships in the clonal hemopathies. When applied to myelopoiesis in AML, obvious differences from the normal are not detected. The analysis is consistent with the view that the blast cell population in AML is distinct from the leukemic myelopoiesis occurring within an abnormal clone. A new assay procedure is described for progenitor cells related to blast cell proliferation. Finally, these concepts are used to develop a model for the pathogenesis and cellular characteristics of AML.

Cells, Cultured↗

High death rates: more deaths or earlier deaths?

In the analysis of mortality statistics high age-specific death rates could be interpreted as meaning more deaths (more disease), but they could equally well be interpreted as meaning earlier deaths (death at younger age). The distinction markedly affects the choice of hypotheses that may be advanced to explain variations in person, time and place and the design of subsequent, more detailed field studies to test the hypotheses. Furthermore, the majority of descriptive papers make no comparisons with a control disease and thereby break one of the ground rules of epidemiology. This paper shows how, in the example of the geographical variations within England and Wales of ischaemic heart disease, a control may be simply introduced and that much of the observed variation is not in the proportion who suffer heart disease deaths but is in the age at which deaths occur.

Adult↗

One explanation for the variability of the bacterial suspension test.

Disinfection kinetic studies of sodium dodecyl sulphate, benzalkonium chloride and sodium hypochlorite against Staphylococcus aureus revealed that when a higher inoculum level of Staph. aureus than normal was used (approximately 1 log higher), the efficacy of disinfection was severely attenuated. Kinetic analysis using the Hom model for experiments carried out on tests using 3 x 108 organisms ml-1 were unable to account for the large increase in disinfection power observed when smaller inoculum levels were used. Since the inoculum was the same in every way except for the numbers used, the large variations in the log reduction/time curves could not be explained by a variation in the resistance of the population to the biocide, as identical log reduction-time curves should have resulted. The level of disinfection achieved for a given concentration of biocide was found to be approximately linearly related to the cell number ml-1 of test solution and not to the log number. The variation observed is believed to occur due to intrinsic self-quenching of the biocide by the microbes during the course of the disinfection test. As the level of free biocide decreases, the rate of reaction decreases, giving the tails of the log reduction/time curves. Such intrinsic self-quenching could explain the large variations known to occur in the legally required disinfection suspension tests.

Bacteriological Techniques↗

PET studies of cerebral glucose metabolism in conscious rhesus macaques.

A growing body of evidence suggests that rhesus macaques may be a good model of human brain aging. We used positron emission tomography (PET) and 18F-fluorodeoxyglucose (FDG) to measure regional cerebral metabolic rates for glucose (rCMRglc) in young and aged rhesus macaques to determine if age-related decreases, such as those reported in humans, also occur in macaques. Whereas the aged animals had lower metabolic rates in every brain region studied, the largest differences were in left temporal cortex. The largest differences were also observed in left temporal cortex when relative rCMRglc values were used. Both rCMRglc and relative rCMRglc were marked by substantial individual variation within the aged group. This variation may parallel the variation observed in behavioral studies. Future studies that include both PET and behavioral measures should help determine if there is a relationship between age-related changes in rCMRglc and behavior.

Aging↗

On the value of geometry-based models for left ventricular volumetry in magnetic resonance imaging and electron beam tomography: a Bland-Altman analysis.

OBJECTIVE: Methodological comparison of ellipsoid model-based approaches and Simpson method to evaluate left ventricular volumetric parameters by magnetic resonance (MR) and electron beam tomography (EBT) and analysis of the origin of possible discrepancies. METHODS AND MATERIAL: 100 subjects (87 patients, 13 healthy volunteers) were studied in MR in various cardiac views and EBT long axis view to determine left ventricular volumes and masses by applying (rotational) ellipsoid and Simpson model. Observer variation and method agreement was quantified by means of variance component and Bland-Altman analysis. RESULTS: Simpson approach showed smaller observer variability than all ellipsoid approaches. All geometry-based models gave smaller left ventricular volumes than Simpson approach, the bias in mass determination was minimal. Whereas high correlation coefficients (typically 0.85-0.95) for left ventricular volume and mass measurements indicated satisfying correspondence between methods, large 95% limits of agreement made a transfer of results for single subjects between Simpson and ellipsoid approaches difficult and between different geometry-based models almost impossible. Because 95% limits of agreement and observer variability of geometry-based approaches were of equal order, the latter could be identified as main limiting factor of methodological agreement. CONCLUSION: MR Simpson approach is superior to all ellipsoid model-based approaches, because observer variability is smaller.

Adult↗

Glancing behind virus load variation in HIV-1 infection.

Although the steady-state virus load during HIV-1 infection is remarkably stable within a patient, it displays variation over several orders of magnitude between patients. Despite intensive research, the host and virus factors that are responsible for the observed variation remain poorly understood. Comparison of model predictions with clinical data suggests that most of the variation in steady-state virus load between patients reflects variation of the net rate at which activated CD4 cells are produced.

Genetic Variation↗

Analysis of normal tissue complication probability of the lung using a reliability model.

The volume effect of normal tissues and organs is an important factor for predicting normal tissue complication probability (NTCP) following partial, heterogeneous irradiation of organs at risk, and reducing the late sequela by conformal radiation therapy. We have previously developed a reliability model for calculation of NTCP, assuming a parallel architecture of functional subunits (FSU), where a critical number (k) out of the total number of FSUs (N) must be intact for the organ to maintain its function. Published data on radiation-induced lethal pneumonitis and altered breathing rate following partial volume irradiation of the mouse lung were analysed, and critical fraction and corresponding spatial density distribution of FSUs were estimated using this model. The critical fraction (k/N) seemed to be similar for the two endpoints, and a value of 0.7 was found to provide good fit to the experimental data. The critical fraction did not vary throughout the lung, and variation in volume effect cannot therefore be attributed to heterogeneous tissue architecture. On the other hand, our analysis revealed that the observed variation in volume effect of mouse lung may be attributed to heterogeneous spatial distribution in FSU density or also the spatial variation in inactivation probability of the FSUs.

Animals↗

Formate in urine as a biological indicator of formaldehyde exposure: a review.

The presence of a small amount of endogenously derived formate in human urine is normal; however, formate derived from the metabolism of formaldehyde, several other industrial compounds and some pharmaceuticals may elevate the urine formate concentration above the normally expected values. This elevation in the urine formate concentration presents the possibility of using this as a tool for monitoring exposure to chemicals. Unfortunately, the use of urine formate as a technique for monitoring personal chemical exposure has yet to be evaluated. This review identifies several potentially important variables that could alter the extent to which formate is eliminated through the urine and that could affect the accuracy of using urine formate concentration as an indicator of chemical exposure. Some potentially important confounders that have been identified, but not evaluated adequately, include dietary intake, nutritional status and exposure to cigarette smoke. Furthermore, the metabolism and elimination kinetics have yet to be adequately demonstrated in humans. Without having controlled for potential confounders in previous pharmacokinetic studies, it is unknown whether or not the large range and variation observed in human studies is due to the confounders or to innate individual variability. Given the poor understanding of the normal variation of formate concentration in the urine, the use of it as a biological indicator of chemical exposure becomes questionable. Without appreciable skin penetration, as in this case, the reliance upon air monitoring alone may be more practical. The evidence at this time suggests that the use of urine formate to monitor chemical exposure offers a broad opportunity for investigative research. At the present time, however, the interpretation of urine formate concentration in samples obtained from workers would be difficult.

Animals↗

Model for current-driven edge-localized modes.

Edge-localized modes (ELMs) are cyclic disturbances in the outer region of tokamak plasmas that are influential in determining present and future tokamak performance. In this Letter, we outline an approach to modeling ELMs in which we envisage toroidal peeling modes initiating a Taylor relaxation [Phys. Rev. Lett. 33, 1139 (1974)10.1103/PhysRevLett.33.1139] of a tokamak outer region plasma. Relaxation produces a peeling destabilizing flattened edge current profile and a stabilizing plasma-vacuum current sheet; the balance between the two determines the radial extent of the relaxed region. The model can be used to predict the energy losses due to an ELM and reproduces experimentally observed variations with edge safety factor and plasma collisionality. There is an intrinsic "deterministic scatter" in the model that also accords with observation.

Journal Article↗

Analysis of the serum antibody responses to type 1 and type 2 fimbriae in mice immunized with Actinomyces viscosus T14V.

The antibody responses of inbred mice immunized with Actinomyces viscosus T14V cells were analyzed using enzyme-linked immunoabsorbent assays (ELISAs) for measuring serum antibodies reactive with A. viscosus T14V cells and type 1 and type 2 fimbriae. In A/J mice immunized intraperitoneally on days 0 and 14, the serum antibody responses approached peak levels during d 19-35, and a dose of 10(8) cells/injection elicited optimal responses. Analysis of the responses of three genetically diverse strains of inbred mice indicated striking variations in the amounts of anti-type 1 (6.5-fold) and anti-type 2 (14.3-fold) antibodies elicited. The observed variations in the magnitude of the anti-fimbrial antibody responses are theoretically of sufficient magnitude to account for significant differences between mouse strains in their ability to inhibit adherence of A. viscosus T14V to saliva-coated hydroxyapatite and other bacteria. These studies provide a model with which the effects of variations in anti-fimbrial antibody responses on bacterial adherence may be analyzed.

Actinomyces↗

Ambulatory care sensitive hospitalization rates in the aged Medicare population in Utah, 1990 to 1994: a rural-urban comparison.

The objective of this study is to compare the likelihood of hospitalization for conditions that are related to the adequacy and use of ambulatory health care services for Medicare beneficiaries residing in rural and urban regions in Utah. The Health Care Financing Administration's (HCFA) hospital discharge database (Utah hospitals: 1990 to 1994) was used to estimate hospitalization rates (with adjustment for out-of-state admissions) for ambulatory care sensitive conditions. Population estimates were obtained from HCFA beneficiary files. Regional hospitalization rates were obtained through ZIP code matching of the hospital discharge and beneficiary files. Medicare beneficiaries aged 65 and older residing in Utah during 1990 to 1994 are the subjects for the study. The main outcome measures include age and sex-adjusted hospitalization rates by region for the entire state and rate ratio estimates for nonurban regions. The results of the study show that Medicare beneficiaries residing in two rural-frontier regions were more likely than urban beneficiaries to be hospitalized for ambulatory care sensitive conditions. Rate ratio estimates were greater than 1.4 for both regions during the study period. These findings suggest a pattern of an increased burden of avoidable secondary complications and disease progression among Utah Medicare beneficiaries residing in some rural regions. This increased burden may be the result of limitations in the ambulatory care system, medical care provider supply, and/or beneficiary propensity to seek care. Variation in disease prevalence or hospital use patterns for these conditions also may be responsible for all or part of the observed variation in ambulatory care sensitive admission rates.

Aged↗

Silver halide sensitized gelatin process effects in holographic lenses recorded on Slavich PFG-01 plates.

In this work we study the feasibility of using silver halide sensitized gelatin based on PFG-01 (Slavich) emulsions to construct uniaxial compound lenses. This processing is able to introduce variations in the thickness and refractive index of the emulsion. We prove that these changes are not sufficient to provide the observed variations in Bragg conditions in the reconstruction and that a shear-type effect must exist to explain the performance of processed emulsions. We study the characteristics of a compound lens, obtaining acceptable image quality, good resolution, and the typical field limitation of volume holographic elements.

Journal Article↗

Comparison between image and flow cytometry. A priori factors that influence technique.

Comparisons between flow cytometry (FCM) and image cytometry (ICM) have found a high concordance rate in pancreatic tissue, with some discrepancies between the two procedures. This study utilized 40 cases of chronic pancreatitis, primary pancreatic adenocarcinoma and metastatic pancreatic adenocarcinoma to determine the concordance rate between the two procedures. The reasons for discrepancies were identified and subsequently used to establish methods for a priori determination of which procedure to use. Using the FACSCAN flow cytometer and the CAS 200 on appropriately stained specimens that were disaggregated from 50-micron sections, we achieved a concordance rate of r = .878 (P < .01) after removing outliers. Thirty-one of 40 cases matched DNA content, and 9 cases had discrepant results. These discrepant cases were evaluated with factor analysis, in part because initial observations suggested that the variables evaluated could be combined into unifying concepts. The nine measured variables were compressed into three factors, which accounted for 68% of the variation observed between the two methods. Readily evaluated features, on a case-by-case basis, including tumor/nontumor ratios, accounted for the largest proportion of this variation. These findings suggest that tumor/nontumor cell ratios in hematoxylin-eosin-stained sections may provide adequate a priori information to direct the choice of either FCM or ICM to measure DNA ploidy in pancreatic tissue.

Adenocarcinoma↗

Host nutritive quality and host plant choice in two grass miners: primary roles for primary compounds?

The relationship between host plant choice and plant nutritional quality was investigated in two oligophagous grass miners Chromatomyia milii and C. nigra (Diptera, Agromyzidae). We tested whether host choice is determined by chemically mediated host suitability for offspring performance and/or adult performance. A second goal was to relate the observed variation among the different fitness parameters to quantitative and qualitative variation in foliar food quality. Choice experiments illustrated that both miners discriminated among grass species, and that C. milii has a smaller host range than C. nigra, as observed under natural conditions. Oviposition preference was correlated with adult feeding preference and related adult performance (longevity and fecundity) for both miners. Offspring performance measures (survival and pupal size) of at least C. nigra were more weakly related to host preference. Nearly all variation in adult performance of both miners was explained by foliar protein content, which had a positive effect on adult longevity and fecundity. Pupal size of both miners was positively related to foliar water and amino acid content and negatively related to lignin content. No clear relationship between host chemistry and offspring survival was observed. These observations show that fitness parameters are differentially related to host chemistry. Secondly, they suggest that chemically mediated host suitability for adult performance is an important determinant of host choice in this species. Finally, the results suggest a primary role for foliar protein content in host choice of the study species in general and in shaping the host range of C. milii in particular.

Animals↗

Mechanisms controlling venom expulsion in the western diamondback rattlesnake, Crotalus atrox.

Although many studies have documented variation in the amount of venom expended during bites of venomous snakes, the mechanistic source of this variation remains uncertain. This study used experimental techniques to examine how two different features of the venom delivery system, the muscle surrounding the venom gland (the Compressor Glandulae in the rattlesnake) and the fang sheath, could influence venom flow in the western diamondback rattlesnake, Crotalus atrox. Differential contraction of the Compressor Glandulae explained only approximately 30% of the variation in venom flow. Lifting (compression) of the fang sheath as occurs during a normal strike produced marked increases in venom flow; these changes were closely correlated and exceed in magnitude by almost 10 x those recorded from the Compressor Glandulae alone. These results suggest that variation in these two aspects of the venom delivery system--both in terms of magnitude and temporal patterning--explain most of the observed variation in venom injection. The lack of functional or mechanical links between the Compressor Glandulae and the fang sheath, and the lack of skeletal or smooth muscle within the fang sheath, make it unlikely that variation in venom flow is under direct neural control. Instead, differential venom injection results from differences in the pressurization by the Compressor Glandulae, the gate keeping effects of the fang sheath and enclosed soft-tissue chambers, and by differences in the pressure returned by peripheral resistance of the target tissue.

Animals↗

Profiling of sulfoconjugates in urine by using precursor ion and neutral loss scans in tandem mass spectrometry. Application to the investigation of heavy metal toxicity in rats.

This paper reports a liquid chromatographic/electrospray ionization mass spectrometric (LC/ESI-MS) method for profiling a wide range of structurally different sulfoconjugated compounds in urine and its application to the characterization of biomarkers for heavy metal toxicity in rat urine. Sulfoconjugates were first isolated by solid-phase extraction and the LC separation was performed on a reversed-phase column. Sulfoconjugates were detected in a triple-quadrupole mass spectrometer by simultaneously monitoring constant losses of 80 u (or 80 Th for doubly charged ions), precursors of m/z 80 (SO(3) (-*)) and precursors of m/z 97 (HSO4-). The ESI-MS detection conditions were optimized on dehydroepiandrosterone sulfate and estradiol sulfate and tested on other sulfoconjugates. The analysis of urine samples from humans and rats by using the developed method allowed the detection of about 15 peaks in each mode of detection. It was then applied to the investigation of heavy metal toxicity in rats. Comparative analysis of the chromatographic fingerprints of urine from control and uranium- and cadmium-treated rats showed several variations in the chromatographic pattern of the sulfoconjugates. Diagnostic m/z ratios were confirmed by analyzing individual urine samples and one of the observed variations seemed to be specific to uranium toxicity. The ion responsible for this variation has been identified as 4-ethylphenol sulfate by comparison of its chromatographic retention time and collision-induced dissociation mass spectra (MS(2) and MS(3) performed on a quadrupole ion trap instrument) with those of the synthesized compound.

Animals↗

Tumor measurement in the nude mouse.

In this study, the pitfalls of tumor measurement in the nude mouse were evaluated. Regarding intermethod variation, diameters of subcutaneous tumors in nude mice were expressed as length, area, and volume; tumor weights were also recorded. These measurements were all compared to a reference standard: water displacement volume. Estimates of area and volume derived from caliper measurements correlated well with water displacement volume (r = 0.97 and 0.98, respectively). At necropsy, tumor weight was the most consistent and reproducible reflection of tumor volume (r = 1.0000). Regarding interobserver variation, mean absolute difference among volumes determined by several investigators who measured the same tumors in living animals was determined. This averaged 15% of the mean calculated volume. Regarding intraobserver variation, observers measured four separate masses in nude mice eight times each. The observers were prevented from realizing that the same animals were being repeatedly evaluated. Volumes were compared in order to quantify the average variation that occurs when a single investigator repeatedly measures the same mass. When large masses were measured, this error was 7%; when small masses were measured, the error was 27%. Recommendations are made for future work employing tumor measurement.

Animals↗