bluuebunny/crossref_metadata_2025 huggingface dataset:
Salvage Conservation Laryngeal Surgery after Irradiation Failure for Early Laryngeal Cancer
Authors: Motamed Mehdi , Laccourreye Ollivier , Bradley Patrick J.
Published: March 2006

Objectives: One third of recurrences after radiotherapy for early laryngeal cancer remain localized. Salvage conservation laryngeal surgery, with total laryngectomy held as reserve, is a surgical management option that is arguably underused. The aim of this review is to report the oncologic and functional results of salvage conservation laryngeal surgery, using the external or the endolaryngeal laser approach.

Study Design: Review article.

Methods: A computerized literature search of the Medline database from 1985 to 2005 was performed using the following search strategy: laryngeal neoplasm/AND salvage therapy/. Studies with a sample size less than 10 and an average follow‐up of less than 24 months were excluded from analysis. The oncologic outcome, functional outcome, length of hospitalization, and the frequency of complications were recorded.

Results: The average reported local control rate for recurrent early glottic cancer after radiotherapy salvaged by using the external or the endolaryngeal laser approach is 77% and 65%, respectively. The average reported overall local control rate, including cases that subsequently required total laryngectomy, is 90% and 83%, respectively. The endolaryngeal approach when compared with the extralaryngeal approach does have the advantage of reduced complications, lesser requirement for tracheostomy and nasogastric feeding, and shortened hospitalization time.

Conclusions: Conservation laryngeal surgery is a safe and effective treatment for recurrent localized disease after radiotherapy for early stage glottic cancer. Local control may be achieved without the sacrifice of laryngeal function, and total laryngectomy may be held in reserve as the ultimate option for salvage without compromising ultimate survival significantly.
Altered Expression Profile of Superoxide Dismutase Isoforms in Nasal Polyps from Nonallergic Patients
Authors: Cheng Yuan‐Kai , Hwang Guang‐Yuh , Lin Chia‐Der , Tsai Ming‐Hsui , Tsai Shih‐We , Chang Weng‐Cheng
Published: March 2006

Objective/Hypothesis: Nasal polyposis (NP) is a chronic inflammatory disease of the upper respiratory tract. The pathophysiology is unknown but has been shown to be multifactorial. Free radical‐mediated damage has been implicated in the pathogenesis of NP. Superoxide dismutases (SODs) are the first and the most important line of antioxidant enzyme defense against reactive oxygen species. Moreover, isozymes of the SOD family are critical for modulating the activity of nitric oxide, a gaseous free radical that is believed to play roles in the physiology and pathology of respiratory tracts. However, the expression profile of SOD isoforms in NP remains unclear. We aimed to investigate the expression profile of the SOD isoforms in nasal polyps from nonallergic patients.

Study Design: Prospective study.

Methods: Nasal polyp tissues were obtained from eight nonallergic patients who underwent elective polypectomy; mucosal specimens from the middle turbinates were acquired from eight subjects without NP as control tissues. The expression profile of SOD isoenzymes, SOD1, SOD2, and SOD3, in the nasal tissues was determined by reverse transcription‐polymerase chain reaction (RT‐PCR), enzyme‐linked immunosorbent assay (ELISA), and Western blotting (WB).

Results: NP in all eight of the NP patients manifested as severe or recurrent sinonasal polyposis clinically. The expression pattern of SOD isoenzymes evaluated by RT‐PCR analysis indicated that the mean levels of SOD1 mRNA and, to a greater extent, SOD3 mRNA were higher in polyp tissues than in control tissues. There was no significant difference in the expression levels of SOD2 mRNA between the two groups. The data from ELISA and WB analysis showed that there were increased expressions of SOD1 and SOD3 protein in polyp tissues compared with the control tissues, but there was no difference in the expression of SOD2 protein between the two groups. The results from RT‐PCR, ELISA, and WB were paralleled and revealed that the expressions of SOD1 and, to a greater extent, SOD3 were higher in polyp tissues than in the control group.

Conclusions: The expressions of SOD3 and SOD1 were higher in polyp tissues. These results are consistent with previously reported data and support the hypothesis that there is increased oxidative stress in NP. Our data also suggest that the SODs might be important in the pathogenesis of NP; however, the roles these SOD isoforms, especially SOD3, play in both normal nasal mucosa and NP require further clarification.
Nasal Vaccination With CpG Oligodeoxynucleotide Induces Protective Immunity Against Nontypeable Haemophilus influenzae in the Nasopharynx
Authors: Abe Nobuyuki , Kodama Satoru , Hirano Takashi , Eto Mayumi , Suzuki Masashi
Published: March 2006

Objectives: Nasal vaccination is an effective therapeutic regimen for preventing otitis media. Since cholera toxin (CT) is toxic, an alternative adjuvant is required for the development of a nasal vaccine. The efficacy of CpG oligodeoxynucleotide (ODN) as a mucosal adjuvant was examined.

Methods: Mice were immunized intranasally with P6 protein of nontypeable Haemophilus influenzae (NTHi) and adjuvant, CT, or CpG ODN, and P6‐specific antibody responses were examined. The expression of P6‐specific cytokine mRNA in splenic CD4+ T cells was also determined. In addition, NTHi challenges were performed and the NTHi was quantified in nasal washes.

Results: P6‐specific IgA in nasal wash and serum IgG titers were elevated significantly after nasal immunization. The IgG1/IgG2a ratio in serum from P6+CpG‐immunized mice was less than that of P6+CT‐immunized mice. Although IL‐6 was expression similarly in both groups, IFN‐γ expression was greater in P6+CpG‐immunized mice than in P6+CT‐immunized mice. Enhanced clearance of NTHi from the nasopharynx was also shown equally in both groups.

Conclusion: These results indicate that CpG ODN might be an effective mucosal adjuvant, acting by mechanisms that are different from CT. These findings suggest that nasal vaccination with P6 and CpG ODN might be an effective regimen for the induction of NTHi‐specific protective immunity.
Diagnosis of Unilateral Recurrent Laryngeal Nerve Paralysis: Laryngeal Electromyography, Subjective Rating Scales, Acoustic and Aerodynamic Measures
Authors: Bielamowicz Steven , Stager Sheila V.
Published: March 2006

Objective/Hypothesis: To determine whether specific laryngeal electromyography (LEMG) patterns in patients with unilateral vocal fold paralysis/paresis (UVFP) are related to etiology of injury, time from onset of injury, patient perception of symptom severity, acoustic measures, and laryngeal aerodynamic measures.

Study Design: This is a retrospective review of 75 patients.

Methods: Each patient received LEMG, acoustic and aerodynamic testing, and a subjective rating scale assessment (the Glottal Closure Index). Statistical analysis by groups were performed using both ξ2 and single‐factor analysis of variance testing.

Results: An iatrogenic etiology was associated with poor tone on LEMG (P = .05). Those individuals evaluated after 3 months after onset demonstrated more nascent units, a sign of reinnervation, compared with individuals evaluated before 3 months (P < .02). Individuals with fewer normal motor units on LEMG had significantly higher mean translaryngeal air flows (P = .044). Individuals with poor recruitment had significantly shorter maximum phonation times (P = .034) and higher mean flows (P = .044). Individuals with better laryngeal tone as noted on LEMG had significantly lower mean flows (P = .06).

Conclusions: Specific LEMG patterns are related to the etiology of the UVFP and time course since recurrent laryngeal nerve injury. LEMG appears to reflect vocal fold muscle tone as seen on laryngeal function studies. In combination, these studies provide a cohesive assessment of laryngeal function in patients with UVFP.
In Vitro Model for Intraoperative Adjustments in an Implantable Hearing Aid (MET)
Authors: Jorge Jesus Rodriguez , Pfister Markus , Zenner Hans P. , Zalaman Ilse M. , Maassen Marcus M.
Published: March 2006

Objective: Assessment of the optimal static preloading of Otologics Middle Ear Transducer (MET) Ossicular Stimulator, when coupled to the incus.

Background: The MET Ossicular Stimulator is a partially implantable electromagnetic middle ear hearing device that transmits vibrations to the ossicular chain. The vibration patterns were measured with laser‐Doppler vibrometry.

Study Design: Experimental.

Material: We used three human cadaveric temporal bones (TB) and one MET ossicular stimulator.

Methods: Laser‐Doppler vibrometry was used for the selection of TBs. The cochlea was subsequently extirpated from the posterior side to measure the vibrational patterns (VP) of the footplate. Three TBs with different VP were selected based on data obtained from volunteers with normal hearing (n = 110): one TB with a VP larger than +1 SD, one TB with a VP in the range of ±1 SD, and 1 TB with a VP smaller than −1 SD. Transfer functions were calculated between VP of the measurement points at the coupling rod, umbo, incus, and footplate. The TBs were subsequently defrosted. The MET was implanted and coupled to the ossicular chain. Different coupling loads were measured at the incus, the umbo, and the footplate.

Results: Optimal transfer function between the MET transducer and the oval window was achieved during contact when the coupling rod advanced 0.0625 mm (90° rotation). Additional advances of 0.0625 mm (180° turn = 0.125 mm) resulted in a decreased vibrational amplitude, ranging between 20 and 40 dB below 3 kHz. The lowest linear distortion occurred up to 10 kHz during direct contact without advancing the coupling rod.
Effects of Time of Contact and Concentration of Caustic Agent on Generation of Injuries
Authors: Mattos Gustavo M. , Lopes Daniel Dias , Mamede Rui Celso Martins , Ricz Hilton , Mello‐Filho Francisco V. , Neto José Barbieri
Published: March 2006

Objectives: Solid caustic soda (CS) ingestion levels continue high in Brazil. The aggressiveness of a caustic agent depends, among other factors, on its concentration and time of contact with mucosa. However, the interdependence of these factors in the production of caustic lesion in the esophageal mucosa is not known, especially regarding CS as the strongest corrosive agent. We analyze the effects of concentration and time of contact on the aggressiveness of CS to the esophagus of live animals.

Study Design/Methods: One milliliter of CS at concentrations between 1.83% and 73.33% was applied to rats. The solution was kept in contact with the mucosa for 10 to 120 minutes. Internal and external organ aspects were analyzed and the epithelium, submucosa, muscle layer, and adventitia were analyzed microscopically

Results: Epithelial necrosis was observed at all concentrations. Among the necrotic layers, the submucosa was observed starting at the 7.33% concentration, and the muscular layer and adventitia were observed at 14.66% concentration. Damage to the pulmonary parenchyma and trachea occurred at 33.66% after 10 minutes, and perforation of the esophagus was observed only after 120 minutes. After 10 minutes, important corrosive lesions installed in the esophageal layers, expanding in depth and superficial extension. The use of heparin had no effect on the production of lesions.

Conclusions: Ten minutes were sufficient to provoke necrosis, and longer contact increased the area of necrosis. Solution concentration levels were more important in damage production: 1.83% was sufficient for epithelial necrosis, 7.33% caused submucosal necrosis, and 14.66% muscle and adventitia necrosis; 33.66% solutions caused lung and trachea damage after 10 minutes and esophageal perforation after 120 minutes.
Endolymphatic Duct Status During Middle Fossa Dissection of the Internal Auditory Canal: A Human Temporal Bone Radiographic Study
Authors: Drew Brian R. , Semaan Maroun T. , Hsu Daniel P. , Megerian Cliff A.
Published: March 2006

Objective: Successful hearing preservation after acoustic neuroma resection is sometimes complicated by delayed hearing deterioration. The middle fossa approach appears to offer superior long‐term hearing results when compared to the retrosigmoid surgical approach. The goal of this study is to investigate the hypothesis that internal auditory canal (IAC) drilling during middle fossa acoustic neuroma removal is associated with a lower incidence of endolymphatic duct (ELD) injury, a potential cause of delayed hearing loss (HL) known to accompany retrosigmoid hearing preservation dissection techniques.

Study Design: A human temporal bone anatomic and radiographic study complemented with a literature review.

Methods: Twenty human temporal bones were analyzed with high‐resolution multislice computed tomography (HRMCT) and subjected to standard extended middle fossa IAC dissection with labyrinthine preservation and follow‐up HRMCT for analyses of the ELD.

Results: Zero of 20 (0%) temporal bones were found to have violation of the ELD with preservation of the labyrinthine structures and the endolymphatic sac. Reviews of human and animal studies indicate that injury to the ELD may create endolymphatic hydrops, a known cause of hearing deterioration.

Conclusion: The ELD is not vulnerable to injury during IAC dissection using the middle fossa approach. A previous radiographic study has shown that the ELD is violated in 24% of temporal bones during retrosigmoid dissection of the IAC. These findings support and may help explain other outcome studies that show that long‐term hearing results are superior with the use of the middle fossa approach when compared to results following retrosigmoid dissection.
Minimally Invasive Management of the Mediastinal Parathyroid Adenoma
Authors: Sukumar Mithran S. , Komanapalli Christopher B. , Cohen James I.
Published: March 2006

Introduction/Methods: Traditional surgical management of mediastinal parathyroid adenomas has required mediastinal exploration via sternotomy or thoracotomy. By contrast minimal access approaches to the mediastinum via videoscopic transthoracic or transcervical approaches can offer equivalent visualization, patient safety and decreased patient morbidity. The availability of sternal retractor systems, the rapid parathyroid hormone (PTH) assay for intraoperative confirmation of cure, recurrent laryngeal nerve monitoring technique and video‐assisted thoracic surgery (VATS) instrumentation have made this possible. The purpose of this article is to discuss these approaches and the results with their application in five consecutive patients.

Results: Five consecutive patients underwent mediastinal exploration of which three were performed transcervically and three transthoracically (one patient underwent both procedures). One patient had a negative thoracic exploration; one patient's course was complicated by transient recurrent laryngeal nerve injury which resolved spontaneously.

Conclusion: Transcervical approaches for resection of anterior mediastinal parathyroid adenomas are possible with the use of the Rultract Skyhook™ Retractor without the need for sternotomy. VATS provides excellent visualization of the middle and posterior mediastinum avoiding the morbidity of thoracotomy.
Antimeasles Immunoglobulin G for Serologic Diagnosis of Otosclerotic Hearing Loss
Authors: Karosi Tamás , Kónya József , Petkó Mihály , Szabó László Z. , Pytel József , Jóri József , Sziklai István
Published: March 2006

Hypothesis: Persistent measles virus infection of the otic capsule is suggested to be an etiologic factor in otosclerosis. Otosclerosis is a disease of complex unknown etiology causing progressive conductive and/or sensorineural hearing loss (HL).

Background: Diagnostic methods of otosclerosis are sensitive to ossicular chain fixation with low specificity for otosclerotic stapes ankylosis.

Methods: Nucleic acid was extracted from stapes footplates of clinically stapes fixation patients (N = 213). Measles virus nucleoprotein RNA was amplified by reverse‐transcriptase polymerase chain reaction. Amplification results were correlated to histologic findings in 49 cases. Antimeasles IgG levels of all clinically stapes fixation as well as control sera specimens were measured by enzyme‐linked immunosorbent assay.

Results: Among clinically stapes fixation patients, 141 stapes footplates contained measles virus RNA. Among 49 histologic specimens, viral RNA was detectable only in histologically otosclerotic stapes footplates (n = 35). Histology for virus‐negative footplates (n = 14) excluded otosclerosis. Antimeasles IgG levels were significantly lower in the sera of patients with virus‐positive stapes than in control sera.

Conclusions: Combination of decreased antimeasles IgG serum level and conductive HL has a great specificity and sensitivity as a diagnostic method in the preoperative evaluation of ossicular chain fixations otosclerosis. Low antimeasles IgG level indicates otosclerosis, whereas high level suggests nonotosclerotic ossicular chain fixations. Preoperative elucidation of the cause of a conductive HL may suggest optional medical treatment in preference to surgical methods.
Recurrent candidaemia in a neonate with Hirschsprung's disease: fluconazole resistance and genetic relatedness of eight Candida tropicalis isolates
Authors: Chong Pei Pei , Chieng David Ching-Soo , Low Lee Yean , Hafeez Asma , Shamsudin Mariana Nor , Seow Heng Fong , Ng Kee Peng
Published: March 2006

The incidence of candidaemia among immunocompromised patients in Malaysia is increasing at an alarming rate. Isolation of clinical strains that are resistant to fluconazole has also risen markedly. We report here the repeated isolation of Candida tropicalis from the blood of a neonatal patient with Hirschsprung's disease. In vitro fluconazole susceptibility tests of the eight isolates obtained at different time points showed that seven of the isolates were resistant and one isolate was scored as susceptible dose-dependent. Random amplification of polymorphic DNA fingerprinting of the isolates using three primers and subsequent phylogenetic analysis revealed that these isolates were highly similar strains having minor genetic divergence, with a mean pairwise similarity coefficient of 0·893±0·041. The source of the infectious agent was thought to be the central venous catheter, as culture of its tip produced fluconazole-resistant C. tropicalis. This study demonstrates the utility of applying molecular epidemiology techniques to complement traditional mycological culture and drug susceptibility tests for accurate and appropriate management of recurrent candidaemia and highlights the need for newer antifungals that can combat the emergence of fluconazole-resistant C. tropicalis strains.
Characterization of extended-spectrum β-lactamase (ESBL)-producing Kuwait and UK strains identified by the Vitek system, and subsequent comparison of the Vitek system with other commercial ESBL-testing systems using these strains
Authors: Dashti A. A. , West P. , Paton R. , Amyes S. G. B.
Published: March 2006

Two hundred and fifty-one unique patient isolates of Klebsiella pneumoniae (123), Escherichia coli (114), Klebsiella oxytoca (7), Enterobacter cloacae (5) and Citrobacter freundii (2), flagged as extended-spectrum β-lactamase (ESBL) positive by the Vitek system (GNS-526 card), were collected. These strains were isolated from a variety of clinical specimens submitted to the clinical bacteriology laboratories of the Royal Infirmary of Edinburgh (RIE), Edinburgh, UK (and associated GP practices), Hairmyers Hospital, Glasgow, UK, and the Amiri and Farwania Hospitals, Kuwait. Of the 101 RIE strains tested, 15 E. coli strains were found to be ESBL negative by Etest ESBL strips. On retesting the 15 E. coli strains with the Vitek GNS-532 card, 14 were found to be ESBL negative, despite being originally flagged as ESBL positive. The remaining 236 ESBL-producing strains were also subjected to the double disc-diffusion (DDD) technique for the detection of ESBLs. Of these, two were false negatives by Etest ESBL test strips (using both cefotaxime and ceftazidime strips), and 38 were false negatives by the DDD method. The Etest false-negative ESBL-producing strains of K. pneumoniae were positive by DDD. Technically, the Vitek method was the least demanding method to perform, as it was an integral part of the routine susceptibility test card. Etest strips were reliable, but were the most expensive of all the techniques used. The DDD test, while relatively inexpensive, was technically subjective, and in our hands, seven of the ESBL-positive strains that were confirmed by the other two techniques were not detected. Despite the false-positive ESBL-producing E. coli strains, the Vitek susceptibility card with its integral ESBL test offers the clinical laboratory a valuable and quick option to screen for ESBL-producing Klebsiella spp. and E. coli as part of the routine laboratory methodology.
Identification of biochemically atypical Staphylococcus aureus clinical isolates with three automated identification systems
Authors: Ishii Yoshikazu , Alba Jimena , Maehara Chikako , Murakami Hinako , Matsumoto Tetsuya , Tateda Kazuhiro , Furuya Nobuhiko , Iwata Morihiro , Yamaguchi Keizo
Published: March 2006

Between January and April 2002, a total of 271 strains of Staphylococcus aureus were isolated from clinical specimens at Toho University Omori Hospital, Japan, including 201 (74·2 %) which were identified as meticillin-resistant S. aureus (MRSA). However, 34 (12·5 %) were biochemically atypical, because they did not produce acid on mannitol salt agar or did not agglutinate in Staphaurex testing but were categorized as MRSA by PCR analysis and by antibiotic susceptibility. Three automatic identification systems, AutoScan-4® (Dade Behring), BD Phoenix™ (Becton Dickinson) and Vitek® 2 (bioMérieux), were evaluated by testing these atypical S. aureus isolates. The AutoScan-4® and Phoenix™ systems identified all 34 isolates as S. aureus. Without additional tests such as Staphaurex, observation of colony pigment and haemolysins on sheep blood agar, Vitek® 2 identified only 16 isolates (47·1 %) as S. aureus with good or better confidence levels and misidentified one of the remaining isolates as Staphylococcus chromogenes. This study shows that it is possible to identify these physiologically atypical S. aureus isolates correctly by using the Phoenix™ and AutoScan-4® fully automatic identification systems.
Strain differentiation of Trichophyton rubrum by randomly amplified polymorphic DNA and analysis of rDNA nontranscribed spacer
Authors: Baeza Lilian Cristiane , Matsumoto Marcelo Teruyuki , Almeida Ana Marisa Fusco , Mendes-Giannini Maria José Soares
Published: March 2006

Trichophyton rubrum is the most common pathogen causing dermatophytosis. Molecular strain-typing methods have recently been developed to tackle epidemiological questions and the problem of relapse following treatment. A total of 67 strains of T. rubrum were screened for genetic variation by randomly amplified polymorphic DNA (RAPD) analysis, with two primers, 5′-d[GGTGCGGGAA]-3′ and 5′-d[CCCGTCAGCA]-3′, as well as by subrepeat element analysis of the nontranscribed spacer of rDNA, using the repetitive subelements TRS-1 and TRS-2. A total of 12 individual patterns were recognized with the first primer and 11 with the second. Phylogenetic analysis of the RAPD products showed a high degree of similarity (>90 %) among the epidemiologically related clinical isolates, while the other strains possessed 60 % similarity. Specific amplification of TRS-1 produced three strain-characteristic banding patterns (PCR types); simple patterns representing one copy of TRS-1 and two copies of TRS-2 accounted for around 85 % of all isolates. It is concluded that molecular analysis has important implications for epidemiological studies, and RAPD analysis is especially suitable for molecular typing in T. rubrum.
Detection of virulence determinants in clinical strains of Salmonella enterica serovar Enteritidis and mapping on macrorestriction profiles
Authors: Soto Sara M. , Rodríguez Irene , Rodicio M. Rosario , Vila Jordi , Mendoza M. Carmen
Published: March 2006

A total of 80 strains of Salmonella enterica serovar Enteritidis, causing gastroenteritis (G) or bacteraemia (B), and three control strains (C), were subjected to: (i) detection of 14 chromosomally and 1 plasmid-located virulence genes by PCR, (ii) detection of DNA polymorphisms by XbaI and BlnI PFGE, and cluster analysis, (iii) mapping of the 15 screened sequences on macrorestriction profiles and (iv) comparison of the screening and mapping results with data available for other Salmonella strains. Identical virulence genotypes and very similar macrorestriction profiles were shown by most S. Enteritidis strains. However, a number of B strains belonged to genomic types with polymorphisms affecting fragments carrying (SPI2-slyA), (SPI2-slyA-phoP/Q-agfA), (SPI4 and/or stn) and spvC. The information obtained provides the basis for further studies on the genetic background of virulence and the molecular epidemiology of S. Enteritidis.
Epidemiological relationships among penicillin non-susceptible Streptococcus pneumoniae strains recovered in the Czech Republic
Authors: Žemličková Helena , Melter Oto , Urbášková Pavla
Published: March 2006

Since 1986, penicillin non-susceptible pneumococci (PNSP) have been found in the Czech Republic. As documented by a nationwide study, the proportion of invasive strains with reduced susceptibility to penicillin has fluctuated around 5 % in the past decade. Although the level of resistance to penicillin remains stable, the contribution of different capsular serotypes among the PNSP population varies. Whereas serotype 19A was predominantly associated with penicillin resistance until 1997, serotype 9V became most common among PNSP strains in 1998. In a collection of PNSP strains (n=225) isolated from 2000 to 2002, the most frequent serotype was 9V (n=91, 40·4 %), followed by 19F (n=30, 13·3 %) and 14 (n=25, 11·5 %). PFGE and multilocus sequence typing were used to characterize a set of PNSP of the currently predominant serotypes 9V (n=42), 14 (n=15) and 19F (n=14). The Spain9V-3 clone [sequence type (ST) 156] was responsible for a large proportion (100 % of serotype 9V strains, n=42; 93·3 % of serotype 14 strains, n=14) of the analysed strains. A representative of the Taiwan19F-14 clone (ST 236) was also recovered in the Czech Republic (a single isolate of serotype 19F). These findings confirm the spread of the major penicillin-resistant clones in the Czech Republic.
Cyclospora infection in five immunocompetent patients in a Turkish university hospital
Authors: Sancak Banu , Akyon Yakut , Ergüven Sibel
Published: March 2006

Infection with Cyclospora has been increasingly reported worldwide in both immunocompetent and immunocompromised individuals. Here the cases of five patients infected with Cyclospora cayetanensis, who sought medical care at Hacettepe University in Turkey, are reported. Diarrhoea occurred from five to fifteen times a day in all of these patients, whose ages ranged from 27 to 67 years. All the patients were considered immunocompetent. Identification of C. cayetanensis was made by detection of the oocysts by using a modified acid-fast stain.
Identification of Campylobacter spp. and discrimination from Helicobacter and Arcobacter spp. by direct sequencing of PCR-amplified cpn60 sequences and comparison to cpnDB, a chaperonin reference sequence database
Authors: Hill Janet E. , Paccagnella Ana , Law Kee , Melito Pasquale L. , Woodward David L. , Price Lawrence , Leung Amy H. , Ng Lai-King , Hemmingsen Sean M. , Goh Swee Han
Published: March 2006

A robust method for the identification of Campylobacter spp. based on direct sequencing of PCR-amplified partial cpn60 sequences and comparison of these to a reference database of cpn60 sequences is reported. A total of 53 Campylobacter isolates, representing 15 species, were identified and distinguished from phenotypically similar Helicobacter and Arcobacter strains. Pairwise cpn60 sequence identities between Campylobacter spp. ranged from 71 to 92 %, with most between 71 and 79 %, making discrimination of these species obvious. The method described overcomes limitations of existing PCR-based methods, which require time-consuming and complex post-amplification steps such as the cloning of amplification products. The results of this study demonstrate the potential for use of the reference chaperonin sequence database, cpnDB, as a tool for identification of bacterial isolates based on cpn60 sequences amplified with universal primers.
Identification of an IS6110 insertion site in plcD, the unique phospholipase C gene of Mycobacterium bovis
Authors: Viana-Niero Cristina , Rodriguez Cesar Alejandro Rosales , Bigi Fabiana , Zanini Marcos Santos , Ferreira-Neto José Soares , Cataldi Angel , Leão Sylvia Cardoso
Published: March 2006

The IS6110 repetitive element is present in multiple copies in most Mycobacterium tuberculosis complex bacteria, except for Mycobacterium bovis strains, which usually contain a single copy of IS6110 located on a 1·9 kb PvuII fragment of the direct repeat region. IS6110 transposition can disrupt coding regions and is a major force of genomic variation. In a previous work it was demonstrated that phospholipase C genes are preferential loci for IS6110 transposition in M. tuberculosis clinical strains. Bacterial phospholipase C enzymes participate in pathogenic mechanisms used by different organisms, and have been implicated in intracellular survival, cytolysis and cell-to-cell spread. Four phospholipase C genes (plcA, plcB, plcC and plcD) were detected in the genomes of M. tuberculosis, Mycobacterium africanum, Mycobacterium microti and ‘Mycobacterium canettii’. M. bovis and the vaccine strain M. bovis Bacillus Calmette–Guérin contain only the plcD gene. In the present work, the existence of IS6110 insertions within plcD, the unique phospholipase C gene of M. bovis, has been investigated by PCR, Southern blot hybridization and sequencing analysis. In 18 (7·3 %) of 245 isolates analysed, the plcD gene was interrupted by the insertion of one copy of IS6110, which in all cases was transposed in the same orientation and at the same position, 1 972 894, relative to the genome of M. bovis AF2122/97. These 18 isolates were distributed in 6 different spoligotype patterns and contained 4 to 8 IS6110 copies. In contrast, strains showing an intact plcD gene contained one (87 %), two (9·4 %) or three (2·4 %) IS6110 copies, and only a single isolate (1·2 %) had four IS6110 copies. The implications of plcD gene disruption in M. bovis have not been fully investigated, but no differences in the organ distribution of the disease were detected when animals infected with strains from the same spoligotype patterns bearing plcD : : IS6110 and intact plcD were compared.
The Moraxella bovis RTX toxin locus mbx defines a pathogenicity island
Authors: Hess John F. , Angelos John A.
Published: March 2006

To characterize flanking regions of thembxoperon inMoraxella bovis, DNA surroundingmbxCABDtolCwas sequenced in haemolytic and nonhaemolytic strains ofM. bovis. In two haemolytic strains ofM. bovis, thembxoperon, including the adjacentM. bovis tolCorthologue, was flanked by approximately 700 bp imperfect repeats. Nonhaemolytic strains ofM. bovishad only one or no such repeats, as well as ORFs identical to those flanking the repeats from haemolyticM. bovis. Two nonhaemolytic strains also contained ORFs with deduced amino acid sequence similarity to bacterialaraJgenes. The G+C content of thembxCABDtolCgene region was lower than the flanking regions. The genetic organization and G+C content ofmbxCABDtolCgenes, and flanking repeats in haemolyticM. bovis, as well as the presence or absence of flanking repeats in nonhaemolyticM. bovis, suggests that this RTX operon is located on a mobile genetic element, and supports the designation of this region as a pathogenicity island, which is believed to be the first such element demonstrated inM. bovis.
Determination of the prevalence of lymphatic filariasis among migrant workers in Kuwait by detecting circulating filarial antigen
Authors: Iqbal Jamshaid , Sher Ali
Published: March 2006

The main objective of this study was to determine the prevalence of filarial infection among migrant workers in Kuwait. The study was conducted from April 2000 to November 2003. A total of 1050 migrant workers (>90 % from the Indian subcontinent) from filarial endemic countries and 260 individuals residing in Kuwait as controls (50 healthy Kuwaiti blood donors, 50 microfilaria-negative individuals from endemic areas and 160 patients with other parasitic infections) were screened for filarial infection. All specimens were tested for microfilaraemia by microscopy of nucleopore-filtered blood (NFB) and detection of circulating filarial antigen (CFA) by an immunochromatographic test (ICT) and the TropBio assay. The overall prevalence of filarial antigenaemia was 18·3 % (192 individuals) using the ICT and 20·3 % (213 individuals) using the TropBio assay. Thirty-two cases (3 %) of Wuchereria bancrofti were detected by microscopy and the mean microfilaria count in these cases was 816 microfilariae ml−1. CFA was detected only in two of the 260 control subjects. Statistical analysis to calculate the sensitivity, specificity and prevalence of infection was carried out using maximum-likelihood statistical methods. The overall sensitivity and specificity of the ICT and TropBio assay to detect CFA were comparable. Compared with NFB microscopy, the sensitivity of the ICT was 93·8 % and specificity ranged from 84 to 100 %. The sensitivity and specificity of the TropBio assay were 90·1 and 100 %, respectively. However, the ICT failed to detect CFA in two cases with a microfilarial load of <20 microfilariae ml−1. In conclusion, the prevalence of filarial infection among the migrant workers in Kuwait was 18·3 % as determined by the ICT.
Role of two-component systems in the virulence of Streptococcus pneumoniae
Authors: Paterson G. K. , Blue C. E. , Mitchell T. J.
Published: March 2006

Understanding of how the human pathogen Streptococcus pneumoniae perceives and responds to its environment in the host offers insight into the pathogenesis of disease caused by this important bacterium and the potential for improved interventions. A central role in this environmental response is played by two-component systems (TCSs), which both sense the environment and drive the cellular response. Molecular advances in the form of genome sequencing, signature-tagged mutagenesis, differential fluorescence induction and microarray analysis have yielded considerable progress in the study of these systems in S. pneumoniae. These recent advances are discussed here, focusing in particular on the role of TCSs in the virulence of S. pneumoniae.
Two cases of parotid gland infection with bacteraemia due to meticillin-resistant Staphylococcus aureus
Authors: Enoch D. A. , Karas J. A. , Emery M. M. , Borland C.
Published: March 2006

Parotid gland infection as a source of meticillin-resistant Staphylococcus aureus bacteraemia has been rarely reported. It is predominantly a disease of the elderly and is associated with significant mortality. Two cases are described here that presented over a 6 month history at a district general hospital. Many cases may be preventable with adequate hydration and good oral hygiene, combined with effective infection control.
An Improved Grafting Technique for Mature Arabidopsis Plants Demonstrates Long-Distance Shoot-to-Root Transport of Phytochelatins in Arabidopsis
Authors: Chen Alice , Komives Elizabeth A. , Schroeder Julian I.
Published: March 2006

Phytochelatins (PCs) are peptides that function in heavy-metal chelation and detoxification in plants and fungi. A recent study showed that PCs have the ability to undergo long-distance transport in a root-to-shoot direction in transgenic Arabidopsis (Arabidopsis thaliana). To determine whether long-distance transport of PCs can occur in the opposite direction, from shoots to roots, the wheat (Triticum aestivum) PC synthase (TaPCS1) gene was expressed under the control of a shoot-specific promoter (CAB2) in an Arabidopsis PC-deficient mutant, cad1-3 (CAB2∷TaPCS1/cad1-3). Analyses demonstrated that TaPCS1 is expressed only in shoots and that CAB2∷TaPCS1/cad1-3 lines complement the cadmium (Cd) and arsenic metal sensitivity of cad1-3 shoots. CAB2∷TaPCS1/cad1-3 plants exhibited higher Cd accumulation in roots and lower Cd accumulation in shoots compared to wild type. Fluorescence HPLC coupled to mass spectrometry analyses directly detected PC2 in the roots of CAB2:TaPCS1/cad1-3 but not in cad1-3 controls, suggesting that PC2 is transported over long distances in the shoot-to-root direction. In addition, wild-type shoot tissues were grafted onto PC synthase cad1-3 atpcs2-1 double loss-of-function mutant root tissues. An Arabidopsis grafting technique for mature plants was modified to obtain an 84% success rate, significantly greater than a previous rate of approximately 11%. Fluorescence HPLC-mass spectrometry showed the presence of PC2, PC3, and PC4 in the root tissue of grafts between wild-type shoots and cad1-3 atpcs2-1 double-mutant roots, demonstrating that PCs are transported over long distances from shoots to roots in Arabidopsis.
Identification and Characterization of a Stress-Inducible and a Constitutive Small Heat-Shock Protein Targeted to the Matrix of Plant Peroxisomes
Authors: Ma Changle , Haslbeck Martin , Babujee Lavanya , Jahn Olaf , Reumann Sigrun
Published: March 2006

Small heat-shock proteins (sHsps) are widespread molecular chaperones for which a peroxisomal localization has not yet been reported. The Arabidopsis (Arabidopsis thaliana) genome encodes two sHsps with putative peroxisomal targeting signals type 1 or 2 (PTS1 or PTS2). As demonstrated by double-labeling experiments using full-length fusion proteins with enhanced yellow fluorescent protein and deletion constructs lacking the putative targeting domains, AtHsp15.7 (At5g37670) and AtAcd31.2 (At1g06460) are targeted to the peroxisome matrix by a functional PTS1 (SKL>) and a functional PTS2 (RLx5HF), respectively. The peroxisomal localization of AtAcd31.2 was further confirmed by isolation of leaf peroxisomes from Arabidopsis by two successive sucrose density gradients, protein separation by one- and two-dimensional gel electrophoresis, and mass spectrometric protein identification. When AtHsp15.7 and AtAcd31.2 were heterologously expressed in yeast (Saccharomyces cerevisiae) and directed to the cytosol by deletion of the PTSs, both sHsps were able to complement the morphological phenotype of yeast mutants deficient in the cytosolic homologs ScHsp42 or ScHsp26. According to expression studies by reverse transcription-PCR, AtAcd31.2 is constitutively expressed, whereas AtHsp15.7 is hardly expressed under normal conditions but strongly induced by heat and oxidative stress, the latter of which was triggered by the catalase inhibitor 3-aminotriazole or the herbicide methyl viologen applied by watering of whole plants or infiltration of rosette leaves. Thus, plants are exceptional among eukaryotes in employing sHsps in the peroxisome matrix to prevent unspecific aggregation of partially denatured proteins under both physiological and stress conditions.
Xanthan Induces Plant Susceptibility by Suppressing Callose Deposition
Authors: Yun Maximina H. , Torres Pablo S. , Oirdi Mohamed El , Rigano Luciano A. , Gonzalez-Lamothe Rocio , Marano María Rosa , Castagnaro Atilio P. , Dankert Marcelo A. , Bouarab Kamal , Vojnov Adrián A.
Published: March 2006

Xanthan is the major exopolysaccharide secreted by Xanthomonas spp. Despite its diverse roles in bacterial pathogenesis of plants, little is known about the real implication of this molecule in Xanthomonas pathogenesis. In this study we show that in contrast to Xanthomonas campestris pv campestris strain 8004 (wild type), the xanthan minus mutant (strain 8397) and the mutant strain 8396, which is producing truncated xanthan, fail to cause disease in both Nicotiana benthamiana and Arabidopsis (Arabidopsis thaliana) plants. In contrast to wild type, 8397 and 8396 strains induce callose deposition in N. benthamiana and Arabidopsis plants. Interestingly, treatment with xanthan but not truncated xanthan, suppresses the accumulation of callose and enhances the susceptibility of both N. benthamiana and Arabidopsis plants to 8397 and 8396 mutant strains. Finally, in concordance, we also show that treatment with an inhibitor of callose deposition previous to infection induces susceptibility to 8397 and 8396 strains. Thus, xanthan suppression effect on callose deposition seems to be important for Xanthomonas infectivity.
Mitogen-Activated Protein Kinase Is Involved in Abscisic Acid-Induced Antioxidant Defense and Acts Downstream of Reactive Oxygen Species Production in Leaves of Maize Plants
Authors: Zhang Aying , Jiang Mingyi , Zhang Jianhua , Tan Mingpu , Hu Xiuli
Published: March 2006

The role of mitogen-activated protein kinase (MAPK) in abscisic acid (ABA)-induced antioxidant defense was investigated in leaves of maize (Zea mays) plants. Treatments with ABA or H2O2 induced the activation of a 46-kD MAPK and enhanced the expression of the antioxidant genes CAT1, cAPX, and GR1 and the total activities of the antioxidant enzymes catalase, ascorbate peroxidase, glutathione reductase, and superoxide dismutase. Such enhancements were blocked by pretreatment with several MAPK kinase inhibitors and reactive oxygen species inhibitors or scavengers. Pretreatment with MAPK kinase inhibitors also substantially arrested the ABA-induced H2O2 production after 2 h of ABA treatment, but did not affect the levels of H2O2 within 1 h of ABA treatment. Pretreatment with several inhibitors of protein tyrosine phosphatase, which is believed to be a negative regulator of MAPK, only slightly prevented the ABA-induced H2O2 production, but did not affect the ABA-induced MAPK activation and ABA-enhanced antioxidant defense systems. These results clearly suggest that MAPK but not protein tyrosine phosphatase is involved in the ABA-induced antioxidant defense, and a cross talk between H2O2 production and MAPK activation plays a pivotal role in the ABA signaling. ABA-induced H2O2 production activates MAPK, which in turn induces the expression and the activities of antioxidant enzymes. The activation of MAPK also enhances the H2O2 production, forming a positive feedback loop.
Analyses of Expressed Sequence Tags from Apple
Authors: Newcomb Richard D. , Crowhurst Ross N. , Gleave Andrew P. , Rikkerink Erik H.A. , Allan Andrew C. , Beuning Lesley L. , Bowen Judith H. , Gera Emma , Jamieson Kim R. , Janssen Bart J. , Laing William A. , McArtney Steve , Nain Bhawana , Ross Gavin S. , Snowden Kimberley C. , Souleyre Edwige J.F. , Walton Eric F. , Yauk Yar-Khing
Published: March 2006

The domestic apple (Malus domestica; also known as Malus pumila Mill.) has become a model fruit crop in which to study commercial traits such as disease and pest resistance, grafting, and flavor and health compound biosynthesis. To speed the discovery of genes involved in these traits, develop markers to map genes, and breed new cultivars, we have produced a substantial expressed sequence tag collection from various tissues of apple, focusing on fruit tissues of the cultivar Royal Gala. Over 150,000 expressed sequence tags have been collected from 43 different cDNA libraries representing 34 different tissues and treatments. Clustering of these sequences results in a set of 42,938 nonredundant sequences comprising 17,460 tentative contigs and 25,478 singletons, together representing what we predict are approximately one-half the expressed genes from apple. Many potential molecular markers are abundant in the apple transcripts. Dinucleotide repeats are found in 4,018 nonredundant sequences, mainly in the 5′-untranslated region of the gene, with a bias toward one repeat type (containing AG, 88%) and against another (repeats containing CG, 0.1%). Trinucleotide repeats are most common in the predicted coding regions and do not show a similar degree of sequence bias in their representation. Bi-allelic single-nucleotide polymorphisms are highly abundant with one found, on average, every 706 bp of transcribed DNA. Predictions of the numbers of representatives from protein families indicate the presence of many genes involved in disease resistance and the biosynthesis of flavor and health-associated compounds. Comparisons of some of these gene families with Arabidopsis (Arabidopsis thaliana) suggest instances where there have been duplications in the lineages leading to apple of biosynthetic and regulatory genes that are expressed in fruit. This resource paves the way for a concerted functional genomics effort in this important temperate fruit crop.
Cryptochrome 1 from Brassica napus Is Up-Regulated by Blue Light and Controls Hypocotyl/Stem Growth and Anthocyanin Accumulation
Authors: Chatterjee Mithu , Sharma Pooja , Khurana Jitendra P.
Published: March 2006

Cryptochromes are blue/ultraviolet-A light sensing photoreceptors involved in regulating various growth and developmental responses in plants. Investigations on the structure and functions of cryptochromes in plants have been largely confined to Arabidopsis (Arabidopsis thaliana), tomato (Lycopersicon esculentum), and pea (Pisum sativum). We report here the characterization of the cryptochrome 1 gene from Brassica napus (BnCRY1), an oilseed crop, and its functional validation in transgenics. The predicted BnCRY1 protein sequence shows a high degree of sequence identity (94%) to Arabidopsis CRY1. A semiquantitative reverse transcription-polymerase chain reaction and the western-blot analysis revealed that blue light up-regulates its transcript and protein levels in young seedlings. The BnCRY1 promoter harbors conventional light-responsive cis-acting elements, which presumably impart light activation to the GUS (β-glucuronidase) reporter gene expressed in Arabidopsis. Although the BnCRY1 transcript could be detected in all the tissues examined, its protein was virtually undetectable in mature leaves and the root, indicating a tissue-specific translational control or protein turnover. The antisense-BnCRY1 Brassica transgenic seedlings accumulated negligible levels of CRY1 protein and displayed an elongated hypocotyl when grown under continuous white or blue light (but not under red or far-red light); the accumulation of anthocyanins was also reduced significantly. The adult transformants were also found to be tall when grown under natural light environment in a containment facility without any artificial illumination. These data provide functional evidence for a role of blue light up-regulated cry1 in controlling photomorphogenesis in Brassica species.
Adenylate-Coupled Ion Movement. A Mechanism for the Control of Nodule Permeability to O2 Diffusion
Authors: Wei Hui , Layzell David B.
Published: March 2006

In response to changes in phloem supply, adenylate demand, and oxygen status, legume nodules are known to exercise rapid (seconds to hours) physiological control over their permeability to oxygen diffusion. Diffusion models have attributed this permeability control to the reversible flow of water into or out of intercellular spaces. To test hypotheses on the mechanism of diffusion barrier control, nodulated soybean (Glycine max L. Merr.) plants were exposed to a range of treatments known to alter nodule O2 permeability (i.e. 10% O2, 30% O2, Ar:O2 exposure, and stem girdling) before the nodules were rapidly frozen, freeze dried, and dissected into cortex and central zone (CZ) fractions that were assayed for K, Mg, and Ca ion concentrations. Treatments known to decrease nodule permeability (30% O2, Ar:O2 exposure, and stem girdling) were consistently associated with an increase in the ratio of [K+] in cortex to [K+] in the CZ tissue, whereas the 10% O2 treatment, known to increase nodule permeability, was associated with a decrease in the [K+]cortex:[K+]CZ. When these findings were considered in the light of previous results, a proposed mechanism was developed for the adenylate-coupled movement of ions and water into and out of infected cells as a possible mechanism for diffusion barrier control in legume nodules.