bluuebunny/crossref_metadata_2025 huggingface dataset:
Melatonin: Identification of Sites of Antigonadal Action in Mouse Brain
Authors: Glass J. David , Lynch G. Robert
Published: October 2006

Long-term implants releasing a small quantity of melatonin (45 nanograms per day) were used to determine the brain sites of the hormone's antigonadal action in a photoperiodic species, the white-footed mouse (
Peromyscus leucopus
). Implants in the medial preoptic and supra- and retrochiasmatic areas elicited complete gonadal regression after 7 weeks. Implants in other brain regions had little effect on the animals' reproductive state.
Erythrosine (Red No. 3) and Its Nonspecific Biochemical Actions: What Relation to Behavioral Changes?
Authors: Mailman Richard B. , Ferris Robert M. , Tang Flora L. M. , Vogel Richard A. , Kilts Clinton D. , Lipton Morris A. , Smith Dorothy A. , Mueller Robert A. , Breese George R.
Published: October 2006

Biochemical studies have shown that the ability of erythrosine to inhibit dopamine uptake into brain synaptosomal preparations is dependent on the concentration of tissue present in the assay mixture. Thus, the finding that erythrosine inhibits dopamine uptake (which, if true, would provide a plausible explanation of the Feingold hypothesis of childhood hyperactivity) may simply be an artifact that results from nonspecific interactions with brain membranes. In addition, although erythrosine given parenterally (50 milligrams per kilogram) did not alter locomotor activity of control of 6-hydroxydopamine-treated rats, erythrosine (50 to 300 milligrams per kilogram) attenuated the effect of punishment in a "conflict" paradigm.
Babesia bovis
: Continuous Cultivation in a Microaerophilous Stationary Phase Culture
Authors: Levy Michael G. , Ristic Miodrag
Published: October 2006

The protozoan parasite
Babesia bovis
, a causative agent of bovine babesiosis, has been continuously cultivated in a settled layer of bovine erythrocytes. Lowered oxygen tension within the layer of host erythrocytes results in a darkening of infected cultures and provides a rapid means of evaluating parasite growth. Deprivation of carbon dioxide causes the merozoites to accumulate in the medium rather than invading new erythrocytes. When separated from the culture, these extraerythrocytic parasites retain their infectivity. Parasites produced in vitro are morphologically identical to parasites from the blood of infected cattle and are susceptible to antibabesial drugs.
An Explanation of the Language of a Chimpanzee
Authors: Thompson Claudia R. , Church Russell M.
Published: October 2006

The language behavior of the chimpanzee Lana, previously described by Rumbaugh (1977), can be simulated by a computer model in which the animal selects, depending upon context, one of six stock sentences with fixed and variable elements.
Reference: The Linguistic Essential
Authors: Savage-Rumbaugh E. Sue , Rumbaugh D. M. , Smith S. T. , Lawson J.
Published: October 2006

Three chimpanzees learned to label three edibles as "foods" and three inedibles as "tools." Two chimpanzees could then similarly categorize numerous objects during blind trial 1 tests when shown only objects' names. The language-like skills of the chimpanzee who failed (Lana) illustrates that apes can use symbols in ways that emulate human usage without comprehending their representational function.
High-Affinity [
3
H]Imipramine Binding in Rat Hypothalamus: Association with Uptake of Serotonin But Not of Norepinephrine
Authors: Langer S. Z. , Moret C. , Raisman R. , Dubocovich M. L. , Briley M.
Published: October 2006

Inhibition of the binding of [
3
H]imipramine and inhibition of the uptake of [
3
H]serotonin and [
3
H]norepinephrine by a series of antidepressants and other drugs were studied in the rat hypothalamus. No correlation was found between the potencies of these drugs for the inhibition of [
3
H]imipramine binding and the inhibition of [
3
H]norepinephrine uptake. There was, however, a highly significant correlation between the potencies of these drugs for the inhibition of [
3
H]imipramine binding and the inhibition of [
3
H]serotonin uptake. These results suggest that high-affinity [
3
H]imipramine binding might be associated with the mechanism of serotonin uptake in the brain.
Effects of Vasopressin on Human Memory Functions
Authors: Weingartner Herbert , Gold Philip , Ballenger James C. , Smallberg Sheila A. , Summers Richard , Rubinow David R. , Post Robert M. , Goodwin Frederick K.
Published: October 2006

Arginine vasopressin and a number of its synthetic analogs augment memory functions in experimental animals. One of these analogs, 1-desamino-8-D-arginine vasopressin (DDAVP), influences human learning and memory. Cognitively unimpaired, as well as cognitively impaired adults, treated with DDAVP for a period of several days, learn information more effectively, as measured by the completeness, organization, and consistency (reliability) of recall. DDAVP also appears to reverse partially the retrograde amnesia that follows electroconvulsive treatment.
Electroencephalogram Correlates of Higher Cortical Functions
Authors: Gevins A. S. , Zeitlin G. M. , Doyle J. C. , Yingling C. D. , Schaffer R. E. , Callaway E. , Yeager C. L.
Published: October 2006

By means of two-stage, nonlinear multivariate pattern recognition, electroencephalograms (EEG's) were analyzed during performance of verbal and spatial tasks. Complex scalp distributions of θ-, β-, and, to a lesser extent, α-band spectral intensities discriminated between the two members of a pair of tasks, such as writing sentences and Koh's block design. Small EEG asymmetries were probably attributable to limb movements and other uncontrolled noncognitive aspects of tasks. Significant EEG differences between cognitive tasks were eliminated when controls for inter-task differences in efferent activity, stimulus characteristics, and performance-related factors were introduced. Each controlled task was associated with an approximately 10 percent reduction, as compared with visual fixation, in the magnitude of α- and β-band spectral intensity. This effect occurred bilaterally and was approximately the same over occipital, parietal, and central regions, with some minor difference over the frontal region in the β band. With these controls, no evidence for lateralization of different cognitive functions was found in the EEG.
CD19 of B Cells as a Surrogate Kinase Insert Region to Bind Phosphatidylinositol 3-Kinase
Authors: Tuveson David A. , Carter Robert H. , Soltoff Stephen P. , Fearon Douglas T.
Published: October 2006

Antigen receptors on B and T lymphocytes transduce signals by activating nonreceptor protein tyrosine kinases (PTKs). A family of receptor PTKs contains kinase insert regions with the sequence tyrosine-X-X-methionine (where X is any amino acid) that when phosphorylated mediate the binding and activation of phosphatidylinositol 3-kinase (PI 3-kinase). The CD19 membrane protein of B cells enhances activation through membrane immunoglobulin M (mIgM) and was found to contain a functional analog of the kinase insert region. Ligation of mIgM induced phosphorylation of CD19 and association with PI 3-kinase. Thus, CD19 serves as a surrogate kinase insert region for mIgM by providing the means for PI 3-kinase activation by nonreceptor PTKs.
Germ-Line Gene Modification and Disease Prevention: Some Medical and Ethical Perspectives
Authors: Wivel Nelson A. , Walters LeRoy
Published: October 2006

There has been considerable debate about the ethics of human germ-line gene modification. As a result of recent advances in the micromanipulation of embryos and the laboratory development of transgenic mice, a lively discussion has begun concerning both the technical feasibility and the ethical acceptability of human germ-line modification for the prevention of serious disease. This article summarizes some of the recent research on germ-line gene modification in animal models. Certain monogenic deficiency diseases that ultimately might be candidates for correction by germ-line intervention are identified. Several of the most frequently considered ethical issues relative to human germ-line gene modification are considered in the context of professional ethics, parental responsibility, and public policy. Finally, it is suggested that there is merit in continuing the discussion about human germ-line intervention, so that this technique can be carefully compared with alternative strategies for preventing genetic disease.
Involvement of Platelet-Endothelial Cell Adhesion Molecule-1 in Neutrophil Recruitment in Vivo
Authors: Vaporciyan Ara A. , DeLisser Horace M. , Yan Horng-Chin , Mendiguren Ignacio I. , Thom Stephen R. , Jones Michael L. , Ward Peter A. , Albelda Steven M.
Published: October 2006

During inflammation, neutrophils migrate from the vascular lumen into extravascular sites. In vitro assays have suggested that platelet-endothelial cell adhesion molecule-1 [PECAM-1 (CD31)], a member of the immunoglobulin superfamily, is required for the transmigration of neutrophils across endothelial monolayers. Antibody to human PECAM-1, which cross-reacts with rat PECAM-1, was found to block not only in vivo accumulation of rat neutrophils into the peritoneal cavity and the alveolar compartment of the lung but also neutrophil accumulation in human skin grafts transplanted onto immunodeficient mice. On the basis of these findings in three different models of inflammation, it appears that PECAM-1 is required for neutrophil transmigration in vivo and may thus be a potential therapeutic target.
Functional Requirement of a Site-Specific Ribose Methylation in Ribosomal RNA
Authors: Sirum-Connolly Karen , Mason Thomas L.
Published: October 2006

The product of the
PET56
nuclear gene of
Saccharomyces cerevisiae
was shown to be required for ribose methylation at a universally conserved nucleotide in the peptidyl transferase center of the mitochondrial large ribosomal RNA (21
S
rRNA). Cells reduced in this activity were deficient in formation of functional large subunits of the mitochondrial ribosome. The purified Pet56 protein catalyzed the site-specific formation of 2′-
O
-methylguanosine on in vitro transcripts of both mitochondrial 21
S
rRNA and
Escherichia coli
23
S
rRNA. These results provide evidence for an essential modified nucleotide in rRNA.
Size Limit of Molecules Permeating the Junctional Membrane Channels
Authors: Simpson Ian , Rose Birgit , Loewenstein Werner R.
Published: October 2006

The permeability of the cell-to-cell membrane channels in salivary gland cell junction (
Chironomus thummi
) was probed with fluorescent-labeled amino acids and synthetic or natural peptides. Molecules up to 1200 daltons pass through the channels with velocities depending on molecular size. Molecules of 1900 daltons or greater do not pass. This passage failure seems to reflect the normal size limit for junctional channel permeation; the channels continue to be permeated by the molecules up to 1200 daltons when these are mixed with the nonpermeant molecules. From this size limit a channel diameter of 10 to 14 angstroms is estimated.
Ornithine Decarboxylase May Function as an Initiation Factor for RNA Polymerase I
Authors: Manen Carol-Ann , Russel Diane H.
Published: October 2006

Reports suggest that the activity of RNA polymerase I is modulated by a labile protein with a half-life of 10 to 20 minutes. Ornithine decarboxylase is the only labile protein (half-life, 10 to 20 minutes) that increases in activity prior to increased RNA polymerase I activity. The addition of a small amount of a highly purified ornithine decarboxylase preparation to an RNA polymerase I assay increases the initial rate of the reaction as well as the time for which the assay is linear. The incorporation patterns of
14
C-labeled adenosine triphosphate and
32
P-labeled adenosine triphosphate into RNA indicate that the addition of ornithine decarboxylase to the RNA polymerase assay increases the rate of initiation. This report demonstrates a novel way to purify ornithine decarboxylase by RNA polymerase I affinity chromatography and presents data in support of the hypothesis that the labile protein which modulates RNA polymerase I activity is ornithine decarboxylase.
The Hydrolytic Water Molecule in Trypsin, Revealed by Time-Resolved Laue Crystallography
Authors: Singer Paul T. , Smalås Arne , Carty Robert P. , Mangel Walter F. , Sweet Robert M.
Published: October 2006

Crystals of bovine trypsin were acylated at the reactive residue, serine 195, to form the transiently stable
p
-guanidinobenzoate. Hydrolysis of this species was triggered in the crystals by a jump in pH. The hydrolysis was monitored by three-dimensional Laue crystallography, resulting in three x-ray diffraction structures, all from the same crystal and each representing approximately 5 seconds of x-ray exposure. The structures were analyzed at a nominal resolution of 1.8 angstroms and were of sufficient quality to reproduce subtle features in the electron-density maps for each of the structures. Comparison of the structures before and after the pH jump reveals that a water molecule has positioned itself to attack the acyl group in the initial step of the hydrolysis of this transient intermediate.
The Heart: A Target Organ for Estradiol
Authors: Stumpf Walter E. , Sar Madhabananda , Aumüller Gerhard
Published: October 2006

Autoradiographic studies of rat heart reveal that tritiated estradiol concentrates in cell nuclei of the myocardium of the atria and auricles, similar to the myometrium of the uterus. This suggests that estrogen has a direct effect on atrial myocytes through which its "protective" action may be mediated. Cardiac glycosides that are known to exert estrogen-like effects on classical estrogen target tissues, such as uterine muscle, endometrium, vagina, and mammary gland, probably act on atrial muscle through a genomic, steroid hormone-like mechanism of action.
Vibrotactile Pattern Perception: Extraordinary Observers
Authors: Craig James C.
Published: October 2006

Two sighted people showed a remarkable ability to perceive vibrotactile patterns generated by the Optacon, a reading aid for the blind. These individuals were able to read at very high rates, 70 to 100 words per minute, through their fingertips. Additional testing showed them to be much better than other people at discriminating and recognizing vibrotactile patterns.
Persistence of Hong Kong Influenza Virus Variants in Pigs
Authors: Shortridge K. F. , Webster R. G. , Butterfield W. K. , Campbell C. H.
Published: October 2006

The A/Hong Kong/1/68 (H3N2) influenza virus which has not been isolated from man for several years, was recently isolated from pigs in Hong Kong. Influenza viruses similar to A/Victoria/3/75, which are currently circulating in man, were also isolated from pigs. Both above-mentioned viruses could be transmitted readily from pig to pig in experimental studies. The isolation of influenza viruses similar to A/Hong Kong/68 from pigs in 1976 suggests that pigs may serve as a potential reservoir for future human pandemics as well as a possible source of genetic information for recombination between human and porcine strains of influenza virus.
A Response Regulator Model in a Simple Sensory System
Authors: Koshland Daniel E.
Published: October 2006

Bacterial behavior is shown to be modulated through a simple on-off switching device which directs migration toward favorable conditions and away from unfavorable ones. The behavioral response is controlled by a rudimentary memory which allows the bacteria to sense gradients over time. The memory can be explained by a biochemical system involving a response regulator whose level relative to a threshold controls flagellar function. The level of the response regulator is itself controlled by factors such as enzyme levels and environmental stimuli. The molecular basis of the model appears to be relevant to more complex hormonal and neural signaling systems.
Sickle Hemoglobin Aggregation: A New Class of Inhibitors
Authors: Votano Joseph R. , Gorecki Marian , Rich Alexander
Published: October 2006

A number of tri- and tetrapeptides have been found to inhibit the aggregation and gelation of deoxygenated sickle cell hemoglobin. These inhibitors have hydrophobic phenylalanine residues at one end and hydrogen bonding lysine or arginine side chains at the other end. The backbone is not very specific. The inhibitors do not modify the oxygen carrying properties of hemoglobin. When the inhibitor and sickle hemoglobin are put inside reconstituted cells, the erythrocytes do not sickle upon deoxygenation. Compounds of this type may develop into useful agents in the therapy of sickle cell anemia.
Impaired Regulation of Alveolar Ventilation and the Sudden Infant Death Syndrome
Authors: Shannon D. C. , Kelly D.
Published: October 2006

Infants who subsequently died of sudden infant death syndrome manifest alveolar hypoventilation during quiet sleep and abnormality of ventilatory response to carbon dioxide breathing in comparison to normal infants.
Perilla Ketone: A Potent Lung Toxin from the Mint Plant,
Perilla frutescens
Britton
Authors: Wilson Benjamin J. , Garst John E. , Linnabary Robert D. , Channell Robert B.
Published: October 2006

Perilla ketone, from the essential oil of
Perilla frutescens
, is a potent pulmonary edemagenic agent for laboratory animals and livestock. This finding would account for reported effects of the plant on grazing cattle. The use of perilla in oriental foods and medicinal preparations suggests possible hazards to human health as well.
Neuronal Circadian Rhythm: Phase Shifting by a Protein Synthesis Inhibitor
Authors: Jacklet Jon W.
Published: October 2006

A potent inhibitor of protein synthesis, anisomycin, was applied (10
-6
M
) in 6-hour pulses at specific phases in the circadian rhythm of endogenous compound action potential (CAP) activity recorded from the eye of
Aplysia
in vitro. The phase of the circadian rhythm was systematically advanced or delayed (up to 15 hours) depending on the specific phase at which the pulse was applied. The resultant phase response curve implicates protein synthesis on the eukaryotic ribosome as a fundamental part of the controlling processes that constitutes the circadian clock.
Axon-Sparing Brain Lesioning Technique: The Use of Monosodium-L-Glutamate and Other Amino Acids
Authors: Simson Earl L. , Gold Richard M. , Standish L. J. , Pellett Peter L.
Published: October 2006

Infusions of monosodium-L-glutamate into the rostral hypothalamus, believed to contain neurons mediating satiety, produced persistent hyperphagia and obesity, thus suggesting that a brain lesion had been produced. Similar infusions into the caudal hypothalamus, believed to contain unmyelinated axons of passage that mediate satiety, failed to alter food intake or body weight. Histological examination of the affected tissue confirmed the behavioral evidence that suggests that this technique spares axons but destroys cell bodies. Infusions of several other amino acids also damaged neurons while sparing axons of passage.
Abnormal Visual Resolution in the Siamese Cat
Authors: Blake Randolph , Antoinetti Diane N.
Published: October 2006

When tested behaviorally, Siamese cats display marked differences in contrast sensitivity compared to ordinary cats. Overall sensitivity is depressed, the high-frequency cutoff point is lower, and there is less falloff in sensitivity at low spatial frequencies. Optical factors may contribute to these differences, or they may be attributable to the well-established anatomical abnormalities within the visual system of the Siamese cat.
[D-Ala
2
]-Met-Enkephalinamide: A Potent, Long-Lasting Synthetic Pentapeptide Analgesic
Authors: Pert Candace B. , Pert Agu , Chang Jaw-Kang , Fong Bosco T.
Published: October 2006

[D-Ala
2
]-Met-enkephalinamide (DALA), a synthetic enkephalin analog designed by in vitro analysis, binds to opiate receptors almost as tightly as methionine-enkephalin. Since it is not susceptible to degradation by brain enzymes, low doses (5 to 10 micrograms) cause profound, long-lasting, morphine-like analgesia when microinjected into rat brain.
Cholinergic Stimulation of the Rat Hypothalamus: Effects of Liver Glycogen Synthesis
Authors: Shimazu Takashi , Matsushita Hiroshi , Ishikawa Koichi
Published: October 2006

Cholinergic stimulation of the lateral hypothalamic neurons with intrahypothalamic microinjections of acetylcholine or carbachol caused a marked increase in the content of the active form of glycogen (starch) synthase in the liver. Total activity of the enzyme (active plus inactive forms) was not increased significantly. The lowest effective dose of acetylcholine was 5 × 10
-10
mole, and the optimum dose was 5 × 10
-9
mole. Similar applications of other neurotransmitters, such as norepinephrine, dopamine, serotonin, and γ-aminobutyric acid, did not affect the enzyme's activity.
Perceptual Analysis of Moving Patterns
Authors: Hochberg Julian , Fallon Peter
Published: October 2006

Configurations of moving points are often perceptually analyzed into relative and common vectors that are different from the actual motions. If a movement configuration is abruptly replaced by a test point whose objective velocity continues the apparent (but illusory) course of one of the original points, observers perceive that course as uninterrupted and colinear. This finding provides a quantitative measure of the vector extraction phenomenon and was used to show that neither of the two current models adequately fits that phenomenon.
Development of a PCR Assay Followed by Nonradioactive Hybridization Using Oligonucleotides Covalently Bound to CovaLink NH Microwells for Detection of Four
Plasmodium
Species in Blood Samples from Humans
Authors: Machouart M. , Bigois-Delemotte L. , Ajana F. , Brizion M. , Biava M. F. , Collomb J. , Fortier B.
Published: September 2006

We developed and evaluated a PCR-based assay to detect four
Plasmodium
species in 79 blood samples from 56 travelers returning from areas where malaria is endemic. DNA amplification targeting a small region of the 18S rRNA gene was performed with
Plasmodium
genus-specific primers. The biotinylated PCR products were then identified by PCR-colorimetric Covalink NH microwell plate hybridization (CMPH) using species-specific phosphorylated probes covalently bound to a pretreated polystyrene surface. The results from PCR-CMPH showed high specificity, and for 47 of the 56 patients (84%), microscopy and PCR-CMPH results were in agreement. Discordant results were reevaluated with microscopy examination, other molecular methods, and DNA sequencing. Except for one patient, discrepancies were resolved in favor of PCR-CMPH: three mixed infections were detected, four species identification errors were corrected, and two negative results were shown to be positive. Our results indicate that PCR-CMPH is a simple, rapid, and specific method for malaria diagnosis. It employs stable reagents and inexpensive equipment, making it suitable for routine epidemiological use.
Real-Time PCR Assay for Detection and Quantification of Hepatitis B Virus Genotypes A to G
Authors: Welzel Tania M. , Miley Wendell J. , Parks Thomas L. , Goedert James J. , Whitby Denise , Ortiz-Conde Betty A.
Published: September 2006

The detection and quantification of hepatitis B virus (HBV) DNA play an important role in diagnosing and monitoring HBV infection as well as assessing therapeutic response. The great variability among HBV genotypes and the enormous range of clinical HBV DNA levels present challenges for PCR-based amplification techniques. In this study, we describe the development, evaluation, and validation of a novel real-time PCR assay designed to provide accurate quantification of DNA from all eight HBV genotypes in patient plasma specimens. A computer algorithm was used to design degenerate real-time PCR primers and probes based upon a large number (
n
= 340) of full-length genomic sequences including HBV genotypes A to H from Europe, Africa, Asia, and North and South America. Genotype performance was tested and confirmed using 59 genotype A to G specimens from two commercially available worldwide genotype panels. This assay has a dynamic range of at least 8 log
10
without the need for specimen dilution, good clinical intra- and interassay precision, and excellent correlation with the Bayer Diagnostics VERSANT HBV DNA 3.0 (branched DNA) assay (
r
= 0.93). Probit analysis determined the 95% detection level was 56 IU/ml, corresponding to 11 copies per PCR well. The high sensitivity, wide linear range, good reproducibility, and genotype inclusivity, combined with a small sample volume requirement and low cost, make this novel quantitative HBV real-time PCR assay particularly well suited for application to large clinical and epidemiological studies.
Molecular Phylogeny of
Sporothrix schenckii
Authors: Marimon Rita , Gené Josepa , Cano Josep , Trilles Luciana , Dos Santos Lazéra Márcia , Guarro Josep
Published: September 2006

The pathogenic dimorphic fungus
Sporothrix schenckii
is the agent responsible for sporotrichosis, an important fungal infection with a worldwide distribution. Little is known about the population structure of
S. schenckii
, although recent molecular and phenotypic data seem to demonstrate that different genetic lineages exist within this species. The aim of this study was to determine, by sequence analysis of three protein coding loci (chitin synthase, β-tubulin, and calmodulin), whether this variability is due to species divergence or intraspecific diversity in
S. schenckii
. We included in the analysis 60 isolates (59 of clinical and 1 of environmental origin) of this species from a wide geographical range. DNA sequence data from the three nuclear regions were used in a phylogenetic analysis. The combined analysis of the three loci revealed the presence of three major clades, one grouping all of the European isolates, another with only Brazilian isolates, and the third with isolates from other South American countries and Africa. A total of 14 100% bootstrap-supported nodes were shown, 6 of them representing putative phylogenetic species. Our data also demonstrated that most of these species prevail in different geographical regions.
Evaluation of the New Chromogenic Medium Candida ID 2 for Isolation and Identification of
Candida albicans
and Other Medically Important
Candida
Species
Authors: Eraso Elena , Moragues María D. , Villar-Vidal María , Sahand Ismail H. , González-Gómez Nagore , Pontón José , Quindós Guillermo
Published: September 2006

The usefulness of Candida ID 2 (CAID2) reformulated medium (bioMérieux, France) has been compared with that of the former Candida ID (CAID; bioMérieux), Albicans ID 2 (ALB2; bioMérieux), and CHROMagar Candida (CAC; Chromagar, France) chromogenic media for the isolation and presumptive identification of clinically relevant yeasts. Three hundred forty-five stock strains from culture collections, and 103 fresh isolates from different clinical specimens were evaluated. CAID2 permitted differentiation based on colony color between
Candida albicans
(cobalt blue; sensitivity, 91.7%; specificity, 97.2%) and
Candida dubliniensis
(turquoise blue; sensitivity, 97.9%; specificity, 96.6%).
Candida tropicalis
gave distinguishable pink-bluish colonies in 97.4% of the strains in CAID2 (sensitivity, 97.4%; specificity, 100%); the same proportion was reached in CAC, where colonies were blue-gray (sensitivity, 97.4%; specificity, 98.7%). CAC and CAID2 showed 100% sensitivity values for the identification of
Candida krusei
. However, with CAID2, experience is required to differentiate the downy aspect of the white colonies of
C. krusei
from other white-colony-forming species. The new CAID2 medium is a good candidate to replace CAID and ALB2, and it compares well to CAC for culture and presumptive identification of clinically relevant
Candida
species. CAID2 showed better results than CAC in some aspects, such as quicker growth and color development of colonies from clinical specimens, detection of mixed cultures, and presumptive differentiation between
C. albicans
and
C. dubliniensis
.
Genital
Candida
Species Detected in Samples from Women in Melbourne, Australia, before and after Treatment with Antibiotics
Authors: Pirotta Marie V. , Garland Suzanne M.
Published: September 2006

Vulvovaginal candidiasis (VVC) remains a common cause of morbidity, with three-quarters of women affected during their lifetimes. Use of antibiotics is an acknowledged trigger for VVC, which adversely affects women's physical and emotional health. Knowledge of patterns of genital
Candida
species-level identification is important for management, as
Candida
species other than
Candida albicans
often fail first-line treatment. A community sample of women with no vaginal symptoms, and who were prescribed antibiotics, was recruited into this study, where the incidence of genital colonization by various
Candida
species was documented, as well as symptoms of VVC plus relevant associations, before and after treatment with antibiotics. Self-collected low vaginal swabs were taken prior to and 8 days after completion of antibiotic treatment, and data on various potential risk factors for VVC were collected simultaneously, with complete data being available for 233 participants. Baseline
Candida
species colonization was present in 21% of women (95% confidence intervals [CI], 17% to 27%), rising to 37% (95% CI, 31% to 44%) after antibiotic treatment. The primary species detected for either period was
C. albicans
(73%), with
Candida glabrata
detected in around 20%. Self-assessed proneness to VVC after antibiotic treatment and baseline colonization with
Candida
spp. were significantly associated with symptomatic VVC after antibiotic treatment. For microbiologically proven candidiasis, VVC symptoms had a sensitivity of 57% and a specificity of 91%. When physicians prescribe antibiotics, the history of risk of VVC is one issue that physicians should discuss with women, particularly those who are self-identified as being prone to VVC. Furthermore, we recommend that definitive microbiological diagnoses be made for women with recurrent symptoms or those failing initial treatment, to guide appropriate therapy.
Two Cases of Infections Due to Multidrug-Resistant
Bacteroides fragilis
Group Strains
Authors: Katsandri Anastasia , Papaparaskevas Joseph , Pantazatou Angeliki , Petrikkos Georgios L. , Thomopoulos Georgios , Houhoula Dimitra P. , Avlamis Athina
Published: September 2006

Bacteroides fragilis
group strains are still considered susceptible to most antimicrobial agents used for the treatment of infections caused by anaerobic organisms. We describe two cases of infections due to isolates simultaneously resistant to clindamycin, tetracycline, cefoxitin, piperacillin-tazobactam, and imipenem and, in one of the two cases, to metronidazole. Such infections, although still rare, do exist and tend to complicate treatment.
Comparative Evaluation of Early Diagnosis of Tuberculous Meningitis by Different Assays
Authors: Quan Chao , Lu Chuan-Zhen , Qiao Jian , Xiao Bao-Guo , Li X.
Published: September 2006

Cerebrospinal fluid (CSF) and peripheral blood (PBL) were sampled multiple times from 25 patients with a clinical diagnosis of tuberculous meningitis (TBM) and 49 controls, including 27 patients with other infectious diseases of the central nervous system and 22 patients with other noninfectious neurological diseases. We used an enzyme-linked immunospot assay (ELISPOT) to detect anti-
Mycobacterium bovis
BCG antibody-secreting cells in CSF and PBL, PCR to detect a repeated insertion sequence (IS
6110
) specific for
Mycobacterium tuberculosis
in CSF, and an enzyme-linked immunosorbent assay (ELISA) to detect anti-BCG antibodies in CSF and PBL. In the meantime, culture of CSF from every TBM and control patient was done on Lowenstein-Jensen medium. ELISPOT proved to be the most valuable test, with a sensitivity of 84.0% and a specificity of 91.8%, and showed a sensitivity of 100.0% with the CSF specimens obtained within 4 weeks after the onset of TBM. The numbers of CSF anti-BCG immunoglobulin-secreting cells tested by ELISPOT were even higher in the early phase of TBM and declined while the disease was going on (
P
= 0.008), which allowed an early diagnosis to be made. The sensitivities of PCR and ELISA were only 75.0% and 52.3%, respectively; and the specificities were 93.7% and 91.6%, respectively. Culture of CSF on Lowenstein-Jensen medium was the least sensitive (16%) compared to the sensitivities of the other three assays. Our results demonstrate that the ELISPOT technique is worthy for routine use in the laboratory to support the clinical diagnosis of TBM.
Detection of Cariogenic
Streptococcus mutans
in Extirpated Heart Valve and Atheromatous Plaque Specimens
Authors: Nakano Kazuhiko , Inaba Hiroaki , Nomura Ryota , Nemoto Hirotoshi , Takeda Munehiro , Yoshioka Hideo , Matsue Hajime , Takahashi Toshiki , Taniguchi Kazuhiro , Amano Atsuo , Ooshima Takashi
Published: September 2006

The involvement of oral bacteria in the pathogenesis of cardiovascular diseases has been the focus of attention in many studies, and several periodontal pathogens have been detected in diseased cardiovascular lesions, suggesting relationships between oral microorganisms and cardiovascular diseases. However, no information is available regarding the involvement of cariogenic pathogens such as
Streptococcus mutans
. The presence of oral streptococcal species and periodontitis-related bacteria in 35 heart valve and 27 atheromatous plaque clinical specimens, as well as 32 dental plaque specimens from the same subjects, was analyzed using a PCR method. Furthermore, broad-range PCR with DNA sequencing analysis was employed to identify the bacterial species in those samples.
Streptococcus mutans
was frequently detected in the heart valve (69%) and atheromatous plaque (74%) specimens, while other bacterial species, including those related to periodontitis, were detected with much lower frequencies. The bacterial composition in cardiovascular tissues was found to be markedly distinct from that in dental plaque, with only a limited number of species, including
S. mutans
, in the cardiovascular regions shown to have possibly originated from the oral cavity. Semiquantitative assay results revealed that
S. mutans
was detected in significant quantities in the heart valve (40%) and atheromatous plaque (48%) specimens, whereas the quantities of all other tested bacterial species, including several related to periodontitis, were negligible in the cardiovascular samples. These results indicate that
S. mutans
is a possible causative agent of cardiovascular disease.
Optimization and Validation of Recombinant Serological Tests for African Swine Fever Diagnosis Based on Detection of the p30 Protein Produced inTrichoplusia niLarvae
Authors: Pérez-Filgueira D. M. , González-Camacho F. , Gallardo C. , Resino-Talaván P. , Blanco E. , Gómez-Casado E. , Alonso C. , Escribano J. M.
Published: September 2006

We describe the validation of an enzyme-linked immunosorbent assay (ELISA) and confirmatory immunoblotting assays based on a recombinant p30 protein (p30r) produced in insect larvae using a baculovirus vector. Such validation included the following: (i) the scaling up and standardization of p30r production and the associated immunoassays, (ii) a broad immunological analysis using a large number of samples (a total of 672) from Spain and different African locations, and (iii) the detection of the ASF virus (ASFV)-antibody responses at different times after experimental infection. Yields of p30r reached up to 15% of the total protein recovered from the infected larvae at 3 days postinfection. Serological analysis of samples collected in Spain revealed that the p30r-based ELISA presented similar sensitivity to and higher specificity than the conventional Office International des Epizooties-approved ASFV ELISA. Moreover, the p30r ELISA was more sensitive than the conventional ELISA test in detecting ASFV-specific antibodies in experimentally infected animals at early times postinfection. Both the recombinant and conventional ELISAs presented variable rates of sensitivity and specificity with African samples, apparently related to their geographical origin. Comparative analyses performed on the sequences, predicted structures, and antigenicities of p30 proteins from different Spanish and African isolates suggested that variability among isolates might correlate with changes in antigenicity, thus affecting detection by the p30r ELISA. Our estimations indicate that more than 40,000 ELISA determinations and 2,000 confirmatory immunoblotting tests can be performed with the p30r protein obtained from a single infected larva, making this a feasible and inexpensive strategy for production of serological tests with application in developing countries.
High-Throughput Identification of Clinical Pathogenic Fungi by Hybridization to an Oligonucleotide Microarray
Authors: Huang Aihua , Li Jun-Wen , Shen Zhi-Qiang , Wang Xin-Wei , Jin Min
Published: September 2006

Here we report the development of an oligonucleotide microarray method that can identify fungal pathogens in a single reaction. Specific oligonucleotide probes targeted to internal transcribed spacer 2 were designed and synthesized. Fungal DNA was amplified by universal primers, and the PCR product was hybridized with the oligonucleotide microarray. A series of specific hybridization profiles corresponding to species were obtained. The 122 strains of fungal pathogens, including standard and clinically isolated strains, used to test the specificity, stability, and sensitivity of the microarray system belonged to 20 species representing 8 genera. We found that the microarray system can successfully discriminate among the fungal pathogens to the species level, with high specificity and stability. The sensitivity was 15 pg/ml of DNA. This oligonucleotide microarray system represents a rapid, simple, and reliable alternative to conventional methods of identifying common clinical fungal isolates.
Evaluation of the New VITEK 2 Extended-Spectrum Beta-Lactamase (ESBL) Test for Rapid Detection of ESBL Production in
Enterobacteriaceae
Isolates
Authors: Spanu Teresa , Sanguinetti Maurizio , Tumbarello Mario , D'Inzeo Tiziana , Fiori Barbara , Posteraro Brunella , Santangelo Rosaria , Cauda Roberto , Fadda Giovanni
Published: September 2006

Extended-spectrum beta-lactamases (ESBLs) are a large, rapidly evolving group of enzymes that confer resistance to oxyimino cephalosporins and monobactams and are inhibited by clavulanate. Rapid reliable detection of ESBL production is a prerequisite for successful infection management and for monitoring resistance trends and implementation of intervention strategies. We evaluated the performance of the new VITEK 2 ESBL test system (bioMérieux, Inc, Hazelwood, Mo.) in the identification of ESBL-producing
Enterobacteriaceae
isolates. We examined a total of 1,129 clinically relevant
Enterobacteriaceae
isolates (including 218 that had been previously characterized). The ESBL classification furnished by the VITEK 2 ESBL test system was concordant with that of the comparison method (molecular identification of beta-lactamase genes) for 1,121 (99.3%) of the 1,129 isolates evaluated. ESBL production was correctly detected in 306 of the 312 ESBL-producing organisms (sensitivity, 98.1%; positive predictive value, 99.3%). False-positive results emerged for 2 of the 817 ESBL-negative isolates (specificity, 99.7%; negative predictive value, 99.3%). VITEK 2 ESBL testing took 6 to 13 h (median, 7.5 h; mean ± SD, 8.2 ± 2.39 h). This automated short-incubation system appears to be a rapid and reliable tool for routine identification of ESBL-producing isolates of
Enterobacteriaceae
.
Fluorescence In Situ Hybridization for Rapid Identification of
Achromobacter xylosoxidans
and
Alcaligenes faecalis
Recovered from Cystic Fibrosis Patients
Authors: Wellinghausen Nele , Wirths Beate , Poppert Sven
Published: September 2006

Achromobacter xylosoxidans
is frequently isolated from the respiratory secretions of cystic fibrosis (CF) patients, but identification with biochemical tests is unreliable. We describe fluorescence in situ hybridization assays for the rapid identification of
Achromobacter xylosoxidans
and
Alcaligenes faecalis
. Both assays showed high sensitivities and high specificities with a collection of 155 nonfermenters from CF patients.
Rapid and Sensitive Detection of Single
Cryptosporidium
Oocysts from Archived Glass Slides
Authors: Sunnotel O. , Snelling W. J. , Xiao L. , Moule K. , Moore J. E. , Millar B. Cherie , Dooley J. S. G. , Lowery C. J.
Published: September 2006

In this study we report on the development and application of a novel method for efficiently extracting and detecting single
Cryptosporidium
oocysts from archived glass slides. Laser capture microscopy was used to extract low numbers of oocysts from archived glass slides. Highly sensitive real-time PCR methods were then developed to enable the rapid detection and identification of
Cryptosporidium
oocysts from these samples. The method was applied to fecal smears stained with a variety of standard oocyst stains and water samples. This application, with samples derived from both public health and water service laboratories, highlighted the strong potential of this method to be used as a rapid high-throughput screening tool for the routine monitoring of
Cryptosporidium
and other medically important pathogens from clinical, veterinary, and environmental water samples. Importantly, the application of our protocol could be used to type
Cryptosporidium
and other pathogens from stored archived glass slides in public health and water service laboratories, providing vital epidemiological updates and helping to identify and trace pathogens and their routes of infection and ultimately improve their control.
Comparison of Transcription-Mediated Amplification and PCR Assay Results for Various Genital Specimen Types for Detection of
Mycoplasma genitalium
Authors: Wroblewski Jennifer K. H. , Manhart Lisa E. , Dickey Kathleen A. , Hudspeth Marie K. , Totten Patricia A.
Published: September 2006

Mycoplasma genitalium
is now recognized as a possible cause of several idiopathic sexually transmitted disease (STD) syndromes. However, due to the difficulty of culture of this fastidious bacterium, nucleic acid amplification tests (NAATs) are necessary for its detection in patient specimens. In the current study we compared a newly developed research-only transcription-mediated amplification (TMA) assay (Gen-Probe Incorporated) to our in-house DNA-based PCR assay for detection of
M. genitalium
. The relative performance characteristics of these two NAATs were assessed with genital specimens from 284 women and 352 men reporting to an STD clinic in Seattle, WA. Among the women,
M. genitalium
was detected by the TMA and PCR assays in 36 (13%) and 39 (14%) vaginal swab specimens, respectively (κ = 0.923); 26 (9%) and 23 (8%) cervical swab specimens, respectively (κ = 0.843); and 25 (9%) and 28 (10%) urine specimens, respectively (κ = 0.687). Among the
M. genitalium
-positive women, the relative sensitivities of detection for the TMA and PCR assays were 84% and 91%, respectively, for vaginal swab specimens; 60% and 53%, respectively, for cervical swab specimens; and 58% and 65%, respectively, for urine specimens. By using an infected patient (a woman positive at any site by TMA assay and at any site by PCR) as a proxy for a “gold standard,” the specificities of detection were >99.5% for both the TMA and the PCR assays. Among the men,
M. genitalium
was detected in 24 urine specimens (6.8%) by the TMA assay, 26 (7.4%) urine specimens by PCR assay, and 32 urine specimens (9%) by either test (κ = 0.791). We conclude that the
M. genitalium
TMA and PCR assays are highly specific and that vaginal swab specimens are the most sensitive specimen type for the detection of
M. genitalium
in women.
Nucleotide Sequence-Based Analysis for Determining the Molecular Epidemiology of
Penicillium marneffei
Authors: Lasker Brent A.
Published: September 2006

The dimorphic fungus,
Penicillium marneffei
, is an emerging opportunistic pathogen endemic in Southeast Asia, especially for those with impaired cellular immunity such as human immunodeficiency virus-infected persons. A discriminatory and reproducible method based on the analysis of nucleotide sequences would facilitate epidemiologic investigations of this fungus. Twenty-four clinical or environmental isolates of
P. marneffei
obtained from China, Thailand, and Vietnam were analyzed by nucleotide sequence analysis. A total of 3,803 bp, consisting of eight nuclear gene fragments (transcription factor [AbaA], catalase [CpeA]], homodomain transcription factor [StlA], isocitrate lyase [Icl1], polyaromatic amino acid biosynthesis [PAA], NADH-dependent glutamate synthase [NGS], lovastatin nonaketide synthase [LNS], a cell wall mannoprotein [MP1], and a gene fragment of the cytochrome oxidase subunit 1 gene [COX1] of the
P. marneffei
mitochondrial genome) were amplified by PCR and then sequenced. No polymorphic sites within the Cox1 gene fragment were observed. Likewise, no nucleotide sequence polymorphisms were observed for three gene fragments: StlA, AbaA, and NGS. Seven single-nucleotide polymorphisms were observed for three gene fragments, Icl1, CpeA, and PAA, providing only a low degree of discriminatory power (D = 0.747). In contrast, the gene fragment for an antigenic cell wall glycoprotein, MP1, a useful immunologic marker for infection, was observed to be highly polymorphic with 12 different MP1 types (D = 0.887). Single-nucleotide polymorphisms were observed at 21 different locations in the MP1 gene fragment. Indels of 3, 21, 24, and 42 bp were observed and were in frame for protein translation. The relatively high degree of MP1 polymorphisms suggests the sequence is rapidly evolving in order to evade host immune responses. After all polymorphic gene sequences were combined, a high degree of genetic variation was observed (D = 0.949) for a total of 16 different haploid sequence types with 11 genotypes represented by single isolates. Phylogenetic analysis detected clusters composed of isolates obtained only from China or Thailand, as well as clusters with a combination of isolates from these two countries, indicating some mixing or common descent. Identical sequences were observed for isolates passed in vitro for 8 weeks, suggesting good reproducibility. The low degree of nucleotide diversity in housekeeping and regulatory genes suggests the recent emergence and spread as a species or an evolutionary bottleneck. In summary, multilocus sequence typing demonstrated a high degree of discriminatory power and reproducibility and may provide a robust and reliable adjunct method for genotyping isolates of
P. marneffei
and facilitating interlaboratory comparisons.
Edema Disease Caused by a Clone of
Escherichia coli
O147
Authors: Helgerson Amy F. , Sharma Vijay , Dow Amie M. , Schroeder Regina , Post Karen , Cornick Nancy A.
Published: September 2006

Edema disease is a systemic disease of weaned pigs caused by host-adapted strains of
Escherichia coli
, most commonly belonging to serogroup O138, O139, or O141. In the late 1990s,
E. coli
O147 strains containing the virulence genes
f18
,
sta
,
stb
, and
stx
2
were recovered from outbreaks of edema disease in the United States. Pulsed-field gel electrophoresis (PFGE) was used to determine that the majority of these strains (34/43) were closely related to one another. Subsequent analysis by multilocus restriction typing confirmed the PFGE results and indicated that the cluster of edema disease strains were only distantly related to other
E. coli
O147 strains. Serogrouping of edema disease isolates from the Iowa State University Veterinary Diagnostic laboratory recovered between 1996 and 2000 indicated that 42% belonged to serogroup O147. Our data suggest that these strains may be a common serotype of edema disease-causing
E. coli
in the United States.