bluuebunny/crossref_metadata_2025 huggingface dataset:
Selection of Antibiotics after Incision and Drainage of Peritonsillar Abscesses
Authors: Kieff David A. , Bhattacharyya Neil , Siegel Noah S. , Salman Salah D.
Published: November 2005

Despite the fact that peritonsillar abscess is the most common complication of acute tonsillitis, the treatment of peritonsillar abscess remains controversial. One element of controversy is the choice of antibiotics after drainage of the abscess. In an attempt to assess the effect of antibiotic choice on the treatment of peritonsillar abscess, we conducted a retrospective review of records from patients with peritonsillar abscess treated with incision and drainage. Our review identified 103 patients, comprising two groups: 58 patients treated with broad‐spectrum intravenous antibiotics and 45 patients treated with intravenous penicillin alone. These patients were hospitalized after incision and drainage, and therefore their clinical courses and responses to therapy could be rigorously assessed. Characterization of illness based on patient age, temperature, and white blood cell count revealed similar severity of illness between the two groups. Comparison of clinical outcomes with respect to hours hospitalized (mean 44.3 ± 6.6 and 38.3 ± 7.1 hours, 95% confidence interval, for broad‐spectrum and penicillin groups, respectively) and mean hours febrile (16.9 ± 5.0 and 13.3 ± 4.2 hours, 95% confidence interval) were not statistically significantly different (p = 0.222 and 0.269, respectively) between groups, indicating that broad‐spectrum antibiotics failed to show greater efficacy than penicillin in the treatment of these patients. The microbiologic characteristics of these infections, failures of therapy, and complication rates were similar to those reported in the literature. These results suggest that intravenous penicillin remains an excellent choice for therapy in cases of peritonsillar abscess requiring parenteral antibiotics after drainage.
Teleconsultation in Otolaryngology: Live Versus Store and Forward Consultations
Authors: Sclafani Anthony P. , Heneghan Conor , Ginsburg Jeffrey , Sabini Paul , Stern Jordan , Dolitsky Jay N.
Published: November 2005

OBJECTIVE

To evaluate the relative strengths and weaknesses of interactive and delayed teleconsultations in otolaryngology.

SETTING

Ambulatory clinic at an urban tertiary care facility.

SUBJECTS

Forty‐five adult patients with known or suspected upper aerodigestive tract pathology.

INTERVENTION

Patients were interviewed by an otolaryngology chief resident (CR) using a standardized protocol; the results were presented to a board‐certified otolaryngologist present locally (LBCO) and a remote physician viewing the encounter by video‐conferencing elsewhere in the hospital (RBCO). The CR performed a complete otolaryngologic examination, including fiberoptic nasopharyngolaryngoscopy. The CR and LBCO viewed the examination on a video monitor; the RBCO viewed the same image on the video‐conferencing monitor. Each physician independently recorded findings and rendered a diagnosis. A third board‐certified otolaryngologist, who reviewed the stored data file (text and stored images) in a delayed fashion (DBCO), documented his findings and made a diagnosis.

RESULTS

The CR and LBCO agreed on diagnosis in 92% (36 of 39) of cases. The LBCO and RBCO arrived at the same diagnosis in 29 of 34 (85%) cases. The DBCO agreed with the LBCO for 18 of 28 (64%) diagnoses. Agreement on management recommendations between the LBCO/DBCO pair were also lower than for the LBCO/RBCO pair.

CONCLUSIONS

Both interactive and delayed techniques can be used to provide relatively accurate clinical consultations in otolaryngology. Telemedicine can be applied for subspecialty consultations, screening programs, remote emergency triage, second opinions, and resident education.
Cell Adhesion Molecules in Normal Laryngeal Development
Authors: Soliman Ahmed M. S. , Jones Richard M. , Flint Paul W.
Published: November 2005

Larynges taken from normally developing rats ranging from embryos starting at embryonic day 10 to adults were harvested. Sections of the larynges were prepared with the use of immunohistochemical techniques with primary antibodies directed to the neural cell adhesion molecule; selected sections were exposed to antibodies directed against the embryonic polysialated form of the neural cell adhesion molecule and against L1. Cell adhesion molecules were present throughout embryonic development associated with the recurrent laryngeal nerve. Downregulation of expression was observed after postnatal day 14. We conclude that cell adhesion molecules may play a role in the normal neural development of the larynx in rats.
Model for Evaluating the Effect of Growth Factors on the Larynx
Authors: Walner David L. , Cotton Robin T. , Willging J. Paul , Bove Kevin E. , Toriumi Dean M.
Published: November 2005

Growth factors are proteins that help regulate the inflammatory response and wound healing in tissues. After laryngotracheal surgery, proper wound healing is important in maintaining the reconstructed airway. The application of growth factor to the respiratory mucosa of the larynx and its effect on wound healing within the airway have not been studied. This study was designed to establish a model for the evaluation of wound healing after the application of growth factor to composite respiratory mucosa and cartilage surfaces at the time of laryngotracheoplasty. Forty rabbits underwent anterior cricoid cartilage split with or without the use of a cartilage graft. Platelet‐derived growth factor or a placebo substance was applied to the wound at the time of surgery. This study offers a model for studying wound healing in the airway that is reproducible with limited morbidity. (Otolaryngol Head Neck Surg 1999;120:78‐83.)
Cytochrome P‐450 Inhibition Blocks Bone Resorption In Vitro and In Vivo
Authors: Choo Moo‐Jin , Chole Richard A.
Published: November 2005

Although the cyclooxygenase and lipoxygenase pathways of arachidonic acid metabolism have been shown to lead to osteoclastic resorption, the cytochrome P450 pathway has not been implicated. We investigated the effects of the cytochrome P450 pathway in IL‐1β –induced calcium release from cultured mouse calvaria in vitro in the presence of clotrimazole, a cytochrome P450 inhibitor, or l‐NG‐arginine methyl ester, a nitric oxide synthase inhibitor. Clotrimazole inhibited calcium release in a dose‐dependent manner; however, l‐NG‐arginine methyl ester did not inhibit resorption. These results suggest that cytochrome P450 may be another possible mediator of IL‐1β–induced bone resorption in vitro. In the in vivo portion of the study, clotrimazole was administered in the gerbil model of adaptive bone modeling. Clotrimazole inhibited osteoclast surface; however, it did not reduce the osteoclast number, mean erosion surface per osteoclast, mineralization surface, or mineral‐apposition rate. These results suggest that clotrimazole may inhibit the activation of osteoclasts and that cytochrome P450–dependent enzymes may be related to osteoclast activation in vivo. (Otolaryngol Head Neck Surg 1999;120:84‐91.)
Effects of Otitis Media with Effusion on the Vestibular System in Children
Authors: Koyuncu Mehmet , Saka M. Muhittin , Tanyeri Yücel , Şleşlen Teoman , ünal Recep , Tekat Atilla , Yilmaz Fatih
Published: November 2005

Middle ear effusion has been considered the most common cause of vestibular disturbance in children. However, there have been only a few studies on vestibular disturbance in children with otitis media with effusion. We studied the vestibular systems of 30 children with otitis media with effusion aged 8 to 13 years and compared the results with 15 age‐ and sex‐matched controls. A questionnaire relating to vestibular disturbance was given to patients and their parents. Spontaneous nystagmus and positional nystagmus were recorded by electronystagmography as diagnostic tests of the vestibular system. Romberg's and past‐pointing tests were performed on children with otitis media with effusion and controls. After vestibular tests were completed, myringotomy was performed, and a ventilation tube was inserted. The questionnaire and the vestibular tests were repeated after the operation and during the first month after surgery. Our study showed that there was a history of vestibular disturbance in 33% of children with otitis media with effusion. Electronystagmography and Romberg's test findings demonstrated that 33% of the children had vestibular dysfunction (p >0.05). After myringotomy with ventilation tube insertion, vestibular test results returned to normal, and symptoms related to vestibular disturbance improved. These findings confirm the assumption that middle ear effusion may affect the vestibular system, which can be resolved after myringotomy with ventilation tube insertion. (Otolaryngol Head Neck Surg 1999;120:117‐21.)
Bees encode behaviorally significant spectral relationships in complex scenes to resolve stimulus ambiguity
Authors: Lotto R. Beau , Wicklein Martina
Published: November 2005

Bees, like humans, can continue to see a surface from its color even when the scene's global illuminant changes (which is a phenomenon called color constancy). It is not known, however, whether they can also generate color-constant behavior in more natural complex scenes that are lit by multiple lights simultaneously, conditions in which most computational models of color constancy fail. To test whether they can indeed solve this more complex problem, bumblebees were raised in a highly controlled, yet ecological relevant environment consisting of a matrix of 64 artificial flowers under four spatially distinct lights. As in nature, the bees had no direct access to spectral information about the illuminants or flowers. Furthermore, the background of all of the flowers in the matrix was black, independent of illumination. The stimulus information presented to the bee was, therefore, far more constrained than that normally experienced in nature. And yet, bees learned to identify the rewarded flowers in each differently illuminated region of the matrix, even when the illumination of one of the regions was switched with one the bees had not previously experienced. These results suggest that bees can generate color-constant behavior by encoding empirically significant contrast relationships between statistically dependent, but visually distinct, stimulus elements of scenes.
Inhibition of FLT3 signaling targets DCs to ameliorate autoimmune disease
Authors: Whartenby Katharine A. , Calabresi Peter A. , McCadden Erin , Nguyen Bao , Kardian David , Wang Tianhong , Mosse Claudio , Pardoll Drew M. , Small Donald
Published: November 2005

Autoimmune diseases often result from inappropriate or unregulated activation of autoreactive T cells. Traditional approaches to treatment of autoimmune diseases through immunosuppression have focused on direct inhibition of T cells. In the present study, we examined the targeted inhibition of antigen-presenting cells as a means to downregulate immune responses and treat autoimmune disease. Dendritic cells (DCs) are the central antigen-presenting cells for the initiation of T cell responses, including autoreactive ones. A large portion of DCs are derived from hematopoietic progenitors that express FLT3 receptor (CD135), and stimulation of the receptor via FLT3 ligand eitherin vivoorin vitrois known to drive expansion and differentiation of these progenitors toward a DC phenotype. We hypothesized that inhibition of FLT3 signaling would thus produce an inhibition of DC-induced stimulation of T cells, thereby inhibiting autoimmune responses. To this end, we used small-molecule tyrosine kinase inhibitors targeted against FLT3 and examined the effects on DCs and their role in the promulgation of autoimmune disease. Results of our studies show that inhibition of FLT3 signaling induces apoptosis in both mouse and human DCs, and thus is a potential target for immune suppression. Furthermore, targeted inhibition of FLT3 significantly improved the course of established disease in a model for multiple sclerosis, experimental autoimmune encephalomyelitis, suggesting a potential avenue for treating autoimmune disease.
A yeast two-hybrid knockout strain to explore thioredoxin-interacting proteinsin vivo
Authors: Vignols Florence , Bréhélin Claire , Surdin-Kerjan Yolande , Thomas Dominique , Meyer Yves
Published: November 2005

All organisms contain thioredoxin (TRX), a regulatory thiol:disulfide protein that reduces disulfide bonds in target proteins. Unlike animals and yeast, plants contain numerous TRXs for which no function has been assignedin vivo. Recentin vitroproteomic approaches have opened the way to the identification of >100 TRX putative targets, but of which none of the numerous plant TRXs can be specifically associated. In contrast,in vivomethodologies, including classical yeast two-hybrid (Y2H) systems, failed to reveal the expected high number of TRX targets. Here, we developed a yeast strain named CY306 designed to identify TRX targetsin vivoby a Y2H approach. CY306 contains a GAL4 reporter system but also carries deletions of endogenous genes encoding cytosolic TRXs (TRX1andTRX2) that presumably compete with TRXs introduced as bait. We demonstrate here that, in the CY306 strain, yeast TRX1 and TRX2, as well asArabidopsisTRX introduced as bait, interact with known TRX targets or putative partners such as yeast peroxiredoxins AHP1 and TSA1, whereas the same interactions cannot be detected in classical Y2H strains. Thanks to CY306, we also show that TRXs interact with the phosphoadenosine-5-phosphosulfate (PAPS) reductase MET16 through a conserved cysteine. Moreover, interactions visualized in CY306 are highly specific depending on the TRX and targets tested. CY306 constitutes a relevant genetic system to explore the TRX interactomein vivoand with high specificity, and opens new perspectives in the search for new TRX-interacting proteins by Y2H library screening in organisms with multiple TRXs.
Differential use of two cyclic electron flows around photosystem I for driving CO
2
-concentration mechanism in C
4
photosynthesis
Authors: Takabayashi Atsushi , Kishine Masahiro , Asada Kozi , Endo Tsuyoshi , Sato Fumihiko
Published: November 2005

Whereas linear electron flow (LEF) in photosynthesis produces both ATP and NADPH, the cyclic electron flow (CEF) around photosystem I has been shown to produce only ATP. Two alternative routes have been shown for CEF; NAD(P)H dehydrogenase (NDH)- and ferredoxin:plastoquinone oxidoreductase (FQR)-dependent flows, but their physiological relevance has not been elucidated in detail. Meanwhile, because C
4
photosynthesis requires more ATP than does C
3
photosynthesis to concentrate CO
2
, it has not been clear how the extra ATP is produced. In this study, to elucidate whether CEF contributes to the additional ATP needed in C
4
photosynthesis, we estimated the amounts of PGR5, which participates in FQR-dependent flow, and NDH-H, a subunit of NDH, in four C
4
species. Although the expression profiles of PGR5 did not correlate well with the additional ATP requirement, NDH was greatly expressed in mesophyll cells in the NAD-malic enzyme (ME) species, and in bundle-sheath cells in NADP-ME species, where there is a strong need for ATP in the respective cells. Our results indicate that CEF via NDH plays a central role in driving the CO
2
-concentrating mechanism in C
4
photosynthesis.
Revealing the large-scale network organization of growth hormone-secreting cells
Authors: Bonnefont Xavier , Lacampagne Alain , Sanchez-Hormigo Angela , Fino Elodie , Creff Audrey , Mathieu Marie-Noelle , Smallwood Sébastien , Carmignac Danielle , Fontanaud Pierre , Travo Pierre , Alonso Gérard , Courtois-Coutry Nathalie , Pincus Steve M. , Robinson Iain C. A. F. , Mollard Patrice
Published: November 2005

Pituitary growth hormone (GH)-secreting cells regulate growth and metabolism in animals and humans. To secrete highly ordered GH pulses (up to 1,000-fold rise in hormone levelsin vivo), the pituitary GH cell population needs to mount coordinated responses to GH secretagogues, yet GH cells display an apparently heterogeneous scattered distribution in 2D histological studies. To address this paradox, we analyzed in 3D both positioning and signaling of GH cells using reconstructive, two-photon excitation microscopy to image the entire pituitary in GH-EGFP transgenic mice. Our results unveiled a homologous continuum of GH cells connected by adherens junctions that wired the whole gland and exhibited the three primary features of biological networks: robustness of architecture across lifespan, modularity correlated with pituitary GH contents and body growth, and connectivity with spatially stereotyped motifs of cell synchronization coordinating cell activity. These findings change our view of GH cells, from a collection of dispersed cells to a geometrically connected homotypic network of cells whose local morphology and connectivity can vary, to alter the timing of cellular responses to promote more coordinated pulsatile secretion. This large-scale 3D view of cell functioning provides a powerful approach to identify and understand other networks of endocrine cells that are thought to be scatteredin situ. Many dispersed endocrine systems exhibit pulsatile outputs. We suggest that cell positioning and associated cell-cell connection mechanisms will be critical parameters that determine how well such systems can deliver a coordinated secretory pulse of hormone to their target tissues.
Identification and Functional Characterization ofArabidopsisPEROXIN4 and the Interacting Protein PEROXIN22[W]
Authors: Zolman Bethany K. , Monroe-Augustus Melanie , Silva Illeana D. , Bartel Bonnie
Published: November 2005

Peroxins are genetically defined as proteins necessary for peroxisome biogenesis. By screening for reduced response to indole-3-butyric acid, which is metabolized to active auxin in peroxisomes, we isolated an Arabidopsis thaliana  peroxin4 (pex4) mutant. This mutant displays sucrose-dependent seedling development and reduced lateral root production, characteristics of plant peroxisome malfunction. We used yeast two-hybrid analysis to determine that PEX4, an apparent ubiquitin-conjugating enzyme, interacts with a previously unidentified Arabidopsis protein, PEX22. A pex4 pex22 double mutant enhanced pex4 defects, confirming that PEX22 is a peroxin. Expression of both Arabidopsis genes together complemented yeast pex4 or pex22 mutant defects, whereas expression of either gene individually failed to rescue the corresponding yeast mutant. Therefore, it is likely that the Arabidopsis proteins can function similarly to the yeast PEX4–PEX22 complex, with PEX4 ubiquitinating substrates and PEX22 tethering PEX4 to the peroxisome. However, the severe sucrose dependence of the pex4 pex22 mutant is not accompanied by correspondingly strong defects in peroxisomal matrix protein import, suggesting that this peroxin pair may have novel plant targets in addition to those important in fungi. Isocitrate lyase is stabilized in pex4 pex22, indicating that PEX4 and PEX22 may be important during the remodeling of peroxisome matrix contents as glyoxysomes transition to leaf peroxisomes.
Suppression of the Floral ActivatorHd3aIs the Principal Cause of the Night Break Effect in Rice[W]
Authors: Ishikawa Ryo , Tamaki Shojiro , Yokoi Shuji , Inagaki Noritoshi , Shinomura Tomoko , Takano Makoto , Shimamoto Ko
Published: November 2005

A short exposure to light in the middle of the night causes inhibition of flowering in short-day plants. This phenomenon is called night break (NB) and has been used extensively as a tool to study the photoperiodic control of flowering for many years. However, at the molecular level, very little is known about this phenomenon. In rice (Oryza sativa), 10 min of light exposure in the middle of a 14-h night caused a clear delay in flowering. A single NB strongly suppressed the mRNA of Hd3a, a homolog of Arabidopsis thaliana FLOWERING LOCUS T (FT), whereas the mRNAs of OsGI and Hd1 were not affected. The NB effect on Hd3a mRNA was maximal in the middle of the 14-h night. The phyB mutation abolished the NB effect on flowering and Hd3a mRNA, indicating that the NB effect was mediated by phytochrome B. Because expression of the other FT-like genes was very low and not appreciably affected by NB, our results strongly suggest that the suppression of Hd3a mRNA is the principal cause of the NB effect on flowering in rice.
Transcription and Histone Modifications in the Recombination-Free Region Spanning a Rice Centromere[W]
Authors: Yan Huihuang , Jin Weiwei , Nagaki Kiyotaka , Tian Shulan , Ouyang Shu , Buell C. Robin , Talbert Paul B. , Henikoff Steven , Jiang Jiming
Published: November 2005

Centromeres are sites of spindle attachment for chromosome segregation. During meiosis, recombination is absent at centromeres and surrounding regions. To understand the molecular basis for recombination suppression, we have comprehensively annotated the 3.5-Mb region that spans a fully sequenced rice centromere. Although transcriptional analysis showed that the 750-kb CENH3-containing core is relatively deficient in genes, the recombination-free region differs little in gene density from flanking regions that recombine. Likewise, the density of transposable elements is similar between the recombination-free region and flanking regions. We also measured levels of histone H4 acetylation and histone H3 methylation at 176 genes within the 3.5-Mb span. Active genes showed enrichment of H4 acetylation and H3K4 dimethylation as expected, including genes within the core. Our inability to detect sequence or histone modification features that distinguish recombination-free regions from flanking regions that recombine suggest that recombination suppression is an epigenetic feature of centromeres maintained by the assembly of CENH3-containing nucleosomes within the core. CENH3-containing centrochromatin does not appear to be distinguished by a unique combination of H3 and H4 modifications. Rather, the varied distribution of histone modifications might reflect the composition and abundance of sequence elements that inhabit centromeric DNA.
Muscle glycogen reduction in man: relationship between surface EMG activity and oxygen uptake kinetics during heavy exercise
Authors: Osborne Mark A. , Schneider Donald A.
Published: November 2005

The purpose of this study was to determine whether muscle glycogen reduction prior to exercise would alter muscle fibre recruitment pattern and change either on‐transient O2uptake () kinetics or theslow component. Eight recreational cyclists (, 55.6 ± 1.3 ml kg−1min−1) were studied during 8 min of heavy constant‐load cycling performed under control conditions (CON) and under conditions of reduced type I muscle glycogen content (GR).was measured breath‐by‐breath for the determination ofkinetics using a double‐exponential model with independent time delays.was higher in the GR trial compared to the CON trial as a result of augmented phase I and II amplitudes, with no difference between trials in the phase II time constant or the magnitude of the slow component. The mean power frequency (MPF) of electromyography activity for the vastus medialis increased over time during both trials, with a greater rate of increase observed in the GR trial compared to the CON trial. The results suggest that the recruitment of additional type II motor units contributed to the slow component in both trials. An increase in fat metabolism and augmented type II motor unit recruitment contributed to the higherin the GR trial. However, the greater rate of increase in the recruitment of type II motor units in the GR trial may not have been of sufficient magnitude to further elevate the slow component whenwas already high and approaching.
Breath‐holding and its breakpoint
Authors: Parkes M. J.
Published: November 2005

This article reviews the basic properties of breath‐holding in humans and the possible causes of the breath at breakpoint. The simplest objective measure of breath‐holding is its duration, but even this is highly variable. Breath‐holding is a voluntary act, but normal subjects appear unable to breath‐hold to unconsciousness. A powerful involuntary mechanism normally overrides voluntary breath‐holding and causes the breath that defines the breakpoint. The occurrence of the breakpoint breath does not appear to be caused solely by a mechanism involving lung or chest shrinkage, partial pressures of blood gases or the carotid arterial chemoreceptors. This is despite the well‐known properties of breath‐hold duration being prolonged by large lung inflations, hyperoxia and hypocapnia and being shortened by the converse manoeuvres and by increased metabolic rate.

Breath‐holding has, however, two much less well‐known but important properties. First, the central respiratory rhythm appears to continue throughout breath‐holding. Humans cannot therefore stop their central respiratory rhythm voluntarily. Instead, they merely suppress expression of their central respiratory rhythm and voluntarily ‘hold’ the chest at a chosen volume, possibly assisted by some tonic diaphragm activity. Second, breath‐hold duration is prolonged by bilateral paralysis of the phrenic or vagus nerves. Possibly the contribution to the breakpoint from stimulation of diaphragm muscle chemoreceptors is greater than has previously been considered. At present there is no simple explanation for the breakpoint that encompasses all these properties.
The otic ganglion in rats and its parotid connection: cholinergic pathways, reflex secretion and a secretory role for the facial nerve
Authors: Khosravani Nina , Sandberg Malin , Ekström J.
Published: November 2005

Otic ganglionectomy in rats was found to have affected the parotid gland more profoundly than section of the auriculotemporal nerve as assesssed by reduction in gland weight (by 33 versus 20%) and total acetylcholine synthesizing capacity (by 88 versus 76%) 1 week postoperatively and, when assessed on the day of surgery under adrenoceptor blockade, by reflex secretion (by 99 versus 88%). The facial nerve contributed to the acetylcholine synthesizing capacity of the gland. Section of the nerve only, at the level of the stylomastoid foramen, reduced the acetylcholine synthesis by 15% and, combined with otic ganglionectomy, by 98% or, combined with section of the auriculotemporal nerve, by 82%. The facial nerve was secretory to the gland, and the response was of a cholinergic nature. The nerve conveyed reflex secretion of saliva and caused secretion of saliva upon stimulation. Most of the facial secretory nerve fibres originated from the otic ganglion, since after otic ganglionectomy (and allowing for nerve degeneration) the secretory response to facial nerve stimulation was markedly reduced (from 23 to 4 μl (5 min)−1). The persisting secetory response after otic ganglionectomy, exaggerated due to sensitization, and the residual acetylcholine synthesizing capacity (mainly depending on the facial nerve) showed that a minor proportion of pre‐ and postganglionic nerve fibres relay outside the otic ganglion. The great auricular nerve, which like the facial nerve penetrates the gland, caused no secretion of saliva upon stimulation. Avulsion of the auriculotemporal nerve was more effective than otic ganglionectomy in reducing the acetylcholine synthesizing capacity (by 94 versus 88%) and as effective as otic ganglionectomy in abolishing reflex secretion (by 99%). When aiming at parasympathetic denervation, avulsion may be the preferable choice, since it is technically easier to perform than otic ganglionectomy.
Use of Biochemical Kinetic Data To Determine Strain Relatedness among
Salmonella enterica
subsp.
enterica
Isolates
Authors: de la Torre E. , Tello M. , Mateu E. M. , Torre E.
Published: November 2005

Classical biotyping characterizes strains by creating biotype profiles that consider only positive and negative results for a predefined set of biochemical tests. This method allows
Salmonella
subspecies to be distinguished but does not allow serotypes and phage types to be distinguished. The objective of this study was to determine the relatedness of isolates belonging to distinct
Salmonella enterica
subsp.
enterica
serotypes by using a refined biotyping process that considers the kinetics at which biochemical reactions take place. Using a Vitek GNI+ card for the identification of gram-negative organisms, we determined the biochemical kinetic reactions (28 biochemical tests) of 135
Salmonella enterica
subsp.
enterica
strains of pig origin collected in Spain from 1997 to 2002 (59
Salmonella
serotype Typhimurium strains, 25
Salmonella
serotype Typhimurium monophasic variant strains, 25
Salmonella
serotype Anatum strains, 12
Salmonella
serotype Tilburg strains, 7
Salmonella
serotype Virchow strains, 6
Salmonella
serotype Choleraesuis strains, and 1
Salmonella enterica
serotype 4,5,12:−:− strain). The results were expressed as the colorimetric and turbidimetric changes (in percent) and were used to enhance the classical biotype profile by adding kinetic categories. A hierarchical cluster analysis was performed by using the enhanced profiles and resulted in 14 clusters. Six major clusters grouped 94% of all isolates with a similarity of ≥95% within any given cluster, and eight clusters contained a single isolate. The six major clusters grouped not only serotypes of the same type but also phenotypic serotype variations into individual clusters. This suggests that metabolic kinetic reaction data from the biochemical tests commonly used for classic
Salmonella enterica
subsp.
enterica
biotyping can possibly be used to determine the relatedness between isolates in an easy and timely manner.
In Vitro Activities of Anidulafungin against More than 2,500 Clinical Isolates of
Candida
spp., Including 315 Isolates Resistant to Fluconazole
Authors: Pfaller M. A. , Boyken L. , Hollis R. J. , Messer S. A. , Tendolkar S. , Diekema D. J.
Published: November 2005

Anidulafungin is an echinocandin antifungal agent with potent activity against
Candida
spp. We assessed the in vitro activity of anidulafungin against 2,235 clinical isolates of
Candida
spp. using the CLSI broth microdilution method. Anidulafungin was very active against
Candida
spp. (the MIC at which 90% of strains are inhibited [MIC
90
] was 2 μg/ml when MIC endpoint criteria of partial inhibition [MIC-2] were used).
Candida albicans
,
C. glabrata
,
C. tropicalis
,
C. krusei
, and
C. kefyr
were the most susceptible species of
Candida
(MIC
90
, 0.06 to 0.12 μg/ml), and
C. parapsilosis
,
C. lusitaniae
, and
C. guilliermondii
were the least susceptible (MIC
90
, 0.5 to 2 μg/ml). In addition, 315 fluconazole-resistant isolates were tested, and 99% were inhibited by ≤1 μg/ml of anidulafungin. These results provide further evidence for the spectrum and potency of anidulafungin activity against a large and geographically diverse collection of clinically important isolates of
Candida
spp.
Rapid Detection of
Mycobacterium tuberculosis
in Respiratory Samples by Transcription-Reverse Transcription Concerted Reaction with an Automated System
Authors: Takakura Shunji , Tsuchiya Shigeo , Isawa Yuichi , Yasukawa Kiyoshi , Hayashi Toshinori , Tomita Motohisa , Suzuki Katsuhiro , Hasegawa Tatsuro , Tagami Takanori , Kurashima Atsuyuki , Ichiyama Satoshi
Published: November 2005

The aim of this study was to evaluate the performance of the transcription-reverse transcription concerted (TRC) method for the detection of
Mycobacterium tuberculosis
complex (MTC) 16S rRNA in clinical respiratory samples for the diagnosis of pulmonary tuberculosis. TRC is a novel method that enables the rapid and the completely homogeneous real-time monitoring of isothermal sequence RNA amplification without any postamplification procedure. The detection limit of the TRC method for MTC was one organism per 100 μl of sputum. The specificity of the method was confirmed by the absence of positive signals for sputum containing 10
6
M. avium
or
M. kansasii
organisms per 100 μl. A total of 201 respiratory samples from patients diagnosed with or suspected of having tuberculosis were tested. Of the 72 MTC culture-positive samples, the TRC method was positive for 52 (sensitivity, 72.2%), whereas the Roche COBAS AMPLICOR PCR was positive for 58 (sensitivity, 80.6%). Both the TRC method and the COBAS AMPLICOR PCR showed no positive identification for any of the 129 culture-negative samples. The percent agreement between the two methods was 95% (191 of 201 samples). The high sensitivity and specificity together with shorter detection time (within 1 h) of the TRC method allow it to be proposed as a useful method for the rapid detection of MTC in respiratory samples.
Clonal Association between
Streptococcus pneumoniae
Serotype 23A, Circulating within the United States, and an Internationally Dispersed Clone of Serotype 23F
Authors: Pai Rekha , Gertz Robert E. , Whitney Cynthia G. , Beall Bernard
Published: November 2005

Streptococcus pneumoniae
is an important pathogen in the United States and is associated with significant morbidity and mortality. Since the introduction of the seven-valent conjugate vaccine, a significant decline in pneumococcal disease has been reported. However, surveillance for pneumococcal disease remains essential, as the extent of cross protection against vaccine-related serotypes is still unclear. Further, any increase in non-vaccine-related serotypes also needs monitoring. We report on a new clonal association between a vaccine-related serotype, serotype 23A, obtained as part of the Active Bacterial Core surveillance, with an established internationally dispersed Pneumococcal Molecular Epidemiology Network (PMEN) clone, clone Colombia
23F
-26. Sixty-two isolates of serotype 23A collected from sterile sites during a 2-year period (2002 and 2003) were characterized. Twenty-one (34%) isolates were penicillin nonsusceptible, although none were fully resistant. Pulsed-field gel electrophoresis and multilocus sequence typing analysis showed that 24 (39%) of the serotype 23A isolates shared either genetic identity or high genetic relatedness with PMEN clone Colombia
23F
-26. Extensive variability was noted within the sequenced region of
pbp2b
in two penicillin-nonsusceptible isolates as well as in PMEN clone Colombia
23F
-26, suggesting that these isolates probably acquired penicillin resistance independently. The emergence of such new serotype and genotype associations highlights the dynamic nature of the pneumococcal population, necessitating continuous monitoring in the post-vaccine era.
Molecular Evolution of Methicillin-Resistant
Staphylococcus aureus
in the Metropolitan Area of Cologne, Germany, from 1984 to 1998
Authors: Wisplinghoff Hilmar , Ewertz Birgitta , Wisplinghoff Susanne , Stefanik Danuta , Plum Georg , Perdreau-Remington Francoise , Seifert Harald
Published: November 2005

To investigate the molecular evolution of methicillin-resistant
Staphylococcus aureus
(MRSA) in a large metropolitan area in Germany, 398 nonrepetitive MRSA isolates recovered from patients from various teaching and nonteaching hospitals in Cologne between 1984 and 1998 were characterized by pulsed-field gel electrophoresis (PFGE). On this basis, 95 representative isolates were selected and further investigated by multilocus sequence typing (MLST),
spa
typing, and staphylococcal cassette chromosome
mec
(SCC
mec
) typing. Overall, there were 9 MLST types and 16
spa
types. The most prevalent sequence types (STs) were ST239 (38% of isolates), ST247 (29%), and ST228 (18%); the most prevalent
spa
types were 37 (32%) and 51 (29%). ST239 comprised five major PFGE types and various unique PFGE patterns, and ST5 comprised two PFGE types. While the same PFGE pattern was not observed among strains with different STs,
spa
type 37 was observed among strains representing two different STs (ST239 and ST241), and these belonged to the same clonal complex as single-locus variants. ST239 was the earliest predominant ST, with the highest prevalence from 1984 to 1988 (96%), followed by ST247 from 1989 to 1993 (83%) and ST228 from 1994 to 1998 (40%).
Spa
type 37 was the most prevalent from 1984 to 1988 (96%),
spa
type 51 was the most prevalent from 1989 to 1993 (83%), and
spa
types 1 and 458 were the most prevalent from 1994 to 1998 (26% and 14%, respectively). The prevalence of SCC
mec
type III decreased from 96% from 1984 to 1988 to 8% from 1989 to 1993, the prevalence of SCC
mec
type I increased from 4% from 1984 to 1988 to 97% from 1989 to 1993 and decreased to 62% from 1994 to 1998. While the genetic diversity of MRSA increased from 1984 to 1998, one prevalent ST usually accounted for most of the isolates in a given time period.
Antigenic Divergence of
Bordetella pertussis
Isolates in Taiwan
Authors: Yao Shu-Man , Lin Yu-Chi , Chou Chen-Ying , Chen Ying-Yan , Hsiao Miao-Ju , Chen Hour-Young , Yan Jer-Jea , Su Hsun-Pi , Li Shu-Ying
Published: November 2005

In recent studies, antigenic divergence has been observed in
Bordetella pertussis
circulating isolates. We collected 80
Bordetella pertussis
isolates in Taiwan from 1998 to 2004 and analyzed them using a combination of pulsed-field gel electrophoresis (PFGE) and sequencing of the
ptxS1
and
prn
genes. The incidence of pertussis increases every 3 years, and most of the isolates prevalent since 1998 have expressed nonvaccine
ptxS1A
and
prn2
alleles. Through PFGE analysis, all isolates could be classified into four major groups, and the incidence of these groups exhibited a correlation with the
prn
allele expressed by the isolates. We found that PFGE is more discriminative than gene sequencing, since it could divide the isolates expressing the
prn2
allele into two groups: one group circulating from 1998 to 2001 and another group circulating from 2001 to 2004. The transition between the two groups in 2000 coincided with an outbreak of 326 cases. This research indicates that the antigenic divergence of
B. pertussis
circulating isolates has evolved over time in Taiwan. Such information will have implications for vaccine policy in Taiwan.
Characterization of a Serodiagnostic Complement Fixation Antigen of
Coccidioides posadasii
Expressed in the Nonpathogenic Fungus
Uncinocarpus reesii
Authors: Yu J.-J. , Kirkland T. N. , Hall L. K. , Wopschall J. , Smith R. C. , Hung C.-Y. , Chen X. , Tarcha E. , Thomas P. W. , Cole G. T.
Published: November 2005

Coccidioides
spp. (
immitis
and
posadasii
) are the causative agents of human coccidioidomycosis. In this study, we developed a novel system to overexpress coccidioidal proteins in a nonpathogenic fungus,
Uncinocarpus reesii
, which is closely related to
Coccidioides
. A promoter derived from the heat shock protein gene (
HSP60
) of
Coccidioides posadasii
was used to control the transcription of the inserted gene in the constructed coccidioidal protein expression vector (pCE). The chitinase gene (
CTS1
) of
C. posadasii
, which encodes the complement fixation antigen, was expressed using this system. The recombinant Cts1 protein (rCts1
Ur
) was induced in pCE-CTS1-transformed
U. reesii
by elevating the cultivation temperature. The isolated rCts1
Ur
showed chitinolytic activity that was identical to that of the native protein and had serodiagnostic efficacy comparable to those of the commercially available antigens in immunodiffusion-complement fixation tests. Using the purified rCts1
Ur
, 74 out of the 77 coccidioidomycosis patients examined (96.1%) were positively identified by enzyme-linked immunosorbent assay. The rCts1
Ur
protein showed higher chitinolytic activity and slightly greater seroreactivity than the bacterially expressed recombinant Cts1. These data suggest that this novel expression system is a useful tool to produce coccidioidal antigens for use as diagnostic antigens.
Cultivable Oral Microbiota of Domestic Dogs
Authors: Elliott David R. , Wilson Michael , Buckley Catherine M. F. , Spratt David A.
Published: November 2005

Bacteria were isolated from the dental plaques of nine dogs and a sample of pooled saliva from five other dogs and were then identified by comparative 16S rRNA gene sequencing. Among 339 isolates, 84 different phylotypes belonging to 37 genera were identified. Approximately half of the phylotypes were identified to the species level, and 28% of these were considered members of the indigenous oral microbiota of humans. The 16S rRNA gene sequences of the remaining 44 phylotypes were not represented in GenBank, and most of these phylotypes were tentatively identified as candidate new species. The genera most frequently isolated from saliva were
Actinomyces
(26%),
Streptococcus
(18%), and
Granulicatella
(17%). The genera most frequently isolated from plaque were
Porphyromonas
(20%),
Actinomyces
(12%), and
Neisseria
(10%). A comparison of the DNA sequences from this study with sequences of the human microbiota available in GenBank showed that, on average, canine and human microbiotas differed by almost 7% in the 16S rRNA gene. In conclusion, this study has shown that the cultivable oral microbiotas of dogs and humans show significant differences.
Detection of Mutations Associated with Isoniazid Resistance in
Mycobacterium tuberculosis
Isolates from China
Authors: Zhang Min , Yue Jun , Yang Yan-ping , Zhang Hong-mei , Lei Jian-qiang , Jin Rui-liang , Zhang Xue-lian , Wang Hong-hai
Published: November 2005

Nine structural genes (
furA
,
katG
,
inhA
,
kasA
, Rv0340,
iniB
,
iniA
,
iniC
, and
efpA
) and two regulatory regions (the
oxyR-ahpC
intergenic region and the promoter of
mabA-inhA
) in 87 isoniazid (INH)-monoresistant and 50 INH-susceptible
Mycobacterium tuberculosis
isolates collected from five provinces of China were analyzed by sequencing. Eighty-two (94.3%) INH-resistant isolates had mutations in the
katG
gene, with the
katG
Ser315Thr mutation predominant (55.2%). No mutation at codon 463 of
katG
was detected among the 50 INH-susceptible isolates with different IS
6110
fingerprints. In addition, there were 35 (40.2%) INH-resistant isolates that had a mutation at codon 463 of
katG
. Of the INH-resistant strains, 20 (23.0%) isolates harbored double mutations at two separate loci of
katG
. Mutations in the
inhA
promoter region occurred in 13 (14.9%) isolates; 4.6% of the isolates had
inhA
structural gene mutations, and 11.5% harbored mutations in the
oxyR-ahpC
intergenic region. Drug resistance-associated mutations were detected in the
iniBAC
region and
efpA
.
Comparison of Real-Time PCR Protocols for Differential Laboratory Diagnosis of Amebiasis
Authors: Qvarnstrom Yvonne , James Cleve , Xayavong Maniphet , Holloway Brian P. , Visvesvara Govinda S. , Sriram Rama , da Silva Alexandre J.
Published: November 2005

Specific identification of
Entamoeba
spp. in clinical specimens is an important confirmatory diagnostic step in the management of patients who may be infected with
Entamoeba histolytica
, the species that causes clinical amebiasis. Distinct real-time PCR protocols have recently been published for identification of
E. histolytica
and differentiation from the morphologically identical nonpathogenic
Entamoeba dispar
. In this study, we compared three
E. histolytica
real-time PCR techniques published by December 2004. The limits of detection and efficiency of each real-time PCR assay were determined using DNA extracted from stool samples spiked with serially diluted cultured
E. histolytica
trophozoites. The ability of each assay to correctly distinguish
E. histolytica
from
E. dispar
was evaluated with DNA extracted from patients' stools and liver aspirates submitted for confirmatory diagnosis. Real-time PCR allowed quantitative analysis of the spiked stool samples, but major differences in detection limits and assay performance were observed among the evaluated tests. These results illustrate the usefulness of comparative evaluations of diagnostic assays.
A Novel Bacterial Mucinase, Glycosulfatase, Is Associated with Bacterial Vaginosis
Authors: Roberton Anthony M. , Wiggins Rebecca , Horner Patrick J. , Greenwood Rosemary , Crowley Theresa , Fernandes Arnold , Berry Monica , Corfield Anthony P.
Published: November 2005

The modifications to the vaginal habitat accompanying a change to vaginal flora in bacterial vaginosis (BV) are poorly understood. In this study enzymes involved in mucin degradation were measured, including a novel glycosulfatase assay. Women attending an emergency walk-in sexually transmitted disease clinic were studied. One high vaginal swab (HVS) was used to prepare a gram-stained smear to determine BV status, using Ison and Hay's criteria, and a separate swab was used for the purposes of the assays. The median glycosulfatase activity was 8.5 (range, −1.2 to 31.9) nmol h
−1
1.5 ml
−1
of HVS suspension in patients with BV compared to 0.5 (range, −0.7 to 9.4) nmol h
−1
1.5 ml
−1
of HVS suspension in patients without BV (
P
= <0.001). The median glycoprotein sialidase activity was 29.2 (range, −17 to 190) nmol h
−1
1.5 ml
−1
of HVS suspension in patients with BV compared to −1.1 (range, −41 to 48) nmol h
−1
1.5 ml
−1
of HVS suspension in patients without BV (
P
< 0.001). A rapid spot test for sialidase was positive in 22/24 patients with BV (sensitivity, 91.7%; 95% confidence interval [CI], 73 to 99%) and negative in 32/35 patients without BV (specificity, 91.4%; 95% CI, 76.9 to 98.2%) (
P
< 0.001). Glycosulfatase activity significantly correlated with both glycoprotein sialidase activity and the sialidase spot test (
P
= 0.006 and
P
< 0.001, respectively). The results are consistent with the hypothesis that the consortium of bacteria present in BV requires the ability to break down mucins in order to colonize the vagina and replace the normal lactobacilli.
Rapid Identification of Human Adenovirus Types 3 and 7 from Respiratory Specimens via Multiplex Type-Specific PCR
Authors: Lee Jin A , Kim Nam Hee , Kim Sun Jung , Choi Eun Hwa , Lee Hoan Jong
Published: November 2005

The rapid diagnosis of human adenovirus (Ad) infection is crucial for the timely recognition of epidemics. Moreover, identification of the serotypes known to cause serious disease can be helpful in therapeutic intervention. A multiplex PCR assay was developed for the rapid detection of adenovirus type 3 (Ad3) and Ad7 directly from clinical specimens. For this assay, three primer pairs (primers were based on the conserved and hypervariable regions of the hexon) were designed in order to simultaneously amplify all adenoviral serotypes and discriminate between Ad3 and Ad7. In our preliminary analysis, this multiplex PCR assay generated amplicons of the consensus primers from all 106 adenoviral isolates of diverse serotypes and proved able to correctly identify Ad3 and Ad7. This assay was subsequently applied to the detection of Ad3 and Ad7 in respiratory specimens. Among the 127 nasal aspirates from which an adenovirus was grown, the sensitivity with which any serotype could be detected was 91% (115/127). Two of the 53 nasal aspirates which did not grow Ads yielded adenovirus-specific bands, which were confirmed by sequencing analysis. Among the 115 specimens which produced common adenoviral bands, the sensitivity with which Ad3 could be detected was 93% (26/28), and the sensitivity with which Ad7 could be detected was 100% (35/35). Five out of the 115 specimens were proved to harbor more than one type of Ad via sequencing analysis of the amplicons, suggesting mixed infection with at least two different serotypes. In conclusion, this multiplex PCR system can be utilized in the rapid identification of Ad3 and Ad7 directly from clinical specimens. Furthermore, this method constitutes a diagnostic strategy for the detection of coinfection by different Ad serotypes.