How reliable is assessment of true vocal cord-arytenoid unit mobility in patients affected by laryngeal cancer? a multi-institutional study on 366 patients from the ARYFIX collaborative group.

PURPOSE: In clinical practice the assessment of the "vocal cord-arytenoid unit" (VCAU) mobility is crucial in the staging, prognosis, and choice of treatment of laryngeal squamous cell carcinoma (LSCC). The aim of the present study was to measure repeatability and reliability of clinical assessment of VCAU mobility and radiologic analysis of posterior laryngeal extension. METHODS: In this multi-institutional retrospective study, patients with LSCC-induced impairment of VCAU mobility who received curative treatment were included; pre-treatment endoscopy and contrast-enhanced imaging were collected and evaluated by raters. According to their evaluations, concordance, number of assigned categories, and inter- and intra-rater agreement were calculated. RESULTS: Twenty-two otorhinolaryngologists evaluated 366 videolaryngoscopies (total evaluations: 2170) and 6 radiologists evaluated 237 imaging studies (total evaluations: 477). The concordance of clinical rating was excellent in only 22.7% of cases. Overall, inter- and intra-rater agreement was weak. Supraglottic cancers and transoral endoscopy were associated with the lowest inter-observer reliability values. Radiologic inter-rater agreement was low and did not vary with imaging technique. Intra-rater reliability of radiologic evaluation was optimal. CONCLUSIONS: The current methods to assess VCAU mobility and posterior extension of LSCC are flawed by weak inter-observer agreement and reliability. Radiologic evaluation was characterized by very high intra-rater agreement, but weak inter-observer reliability. The relevance of VCAU mobility assessment in laryngeal oncology should be re-weighted. Patients affected by LSCC requiring imaging should be referred to dedicated radiologists with experience in head and neck oncology.

Transoral laser surgery for recurrent glottic cancer after radiotherapy: oncologic and functional outcomes.

Primary radiotherapy (RT) has been successfully employed for treatment of early glottic cancer for the past half century. However, local recurrences still occur in 5-35% of patients. Salvage options for persistent/recurrent glottic cancer include total laryngectomy, open neck partial laryngectomies (ONPLs) and transoral laser surgery (TLS). We performed a retrospective chart review of 35 patients with glottic squamous cell carcinoma previously submitted to RT and managed by TLS at the Department of Otorhinolaryngology - Head and Neck Surgery of the University of Brescia, Italy, from 1995 to 2009. Oncologic outcomes were estimated using the Kaplan-Meier method, and separately calculated for the entire cohort of patients (n = 35) and for patients with true persistent/recurrent disease (n = 30), thus excluding the second primary tumours (n = 5). Hospitalization time and complications were obtained by chart review. Functional outcomes of a subgroup of 10 patients in terms of speech and swallowing were analyzed by the Voice Handicap Index (VHI), GRBAS scale, Multi Dimensional Voice Program (MDVP), M.D. Anderson Dysphagia Inventory (MDADI) questionnaire, videoendoscopy (VEES), and videofluoroscopy (VFS) of swallowing (both graded according to Donzelli's scale), and compared to a cohort of 10 patients matched for age, gender and pT category, treated by the same team of surgeons by TLS as a primary treatment. The types of resection used were: 18 Type III, 1 Type IV, and 16 Type V cordectomies. Postoperative staging was 16 rpT1a, 17 rpT2, and 2 rpT3. The 5-year overall survival for the entire series was 91%. Five-year disease-specific survival, local control with laser alone, and organ preservation rates were 94%, 84% and 87%, respectively. Among the variables tested by univariate analysis, for the entire cohort of patients the pT category had a statistically significant impact on local control with laser alone. Anterior transcommissural extension had a borderline statistical impact on disease-specific survival, while it was clearly significant on overall survival. The status of surgical margins and presence of recurrence after TLS statistically influenced both organ preservation and local control with laser alone. The mean values of VHI, MDADI, and MDVP did not show any statistically significant difference between irradiated and non-irradiated patients. The same was true for GRBAS, VEES, and VFS. This series confirms that TLS after RT failure can be considered a successful surgical option in selected early recurrences, with functional outcomes comparable to those observed after TLS as a primary treatment, and much better than those classically described after ONPLs.

Narrow band imaging and high definition television in the endoscopic evaluation of upper aero-digestive tract cancer.

Narrow band imaging and high definition television are recent innovations in upper aero-digestive tract endoscopy. Aim of this prospective, non-randomized, unblinded study was to establish the diagnostic advantage of these procedures in the evaluation of squamous cell cancer arising from various upper aero-digestive tract sites. Between April 2007 and January 2010, 444 patients affected by upper aero-digestive tract squamous cell cancer, or previously treated for it, were evaluated by white light and narrow band imaging ± high definition television endoscopy, both in the pre-/intra-operative setting and during follow-up. Tumour resection was performed taking into account narrow band imaging and high definition television information to obtain histopathologic confirmation of their validity. Endoscopic and pathologic data were subsequently matched to obtain sensitivity, specificity, positive, negative predictive values, and accuracy. Overall, 110 (25%) patients showed adjunctive findings by narrow band imaging ± high definition television when compared to standard white light endoscopy. Of these patients, 98 (89%) received histopatological confirmation. The sensitivity, specificity, positive, negative predictive values, and accuracy for white light-high definition television were 41%, 92%, 87%, 82%, and 67%, for narrow band imaging alone 75%, 87%, 87%, 74%, and 80%, and for narrow band imaging-high definition television 97%, 84%, 88%, 96%, and 92%. The highest diagnostic gain was observed in the oral cavity and oropharynx (25%). Narrow band imaging and high definition television were of value in the definition of superficial tumour extension, and in the detection of synchronous lesions in the pre-/intra-operative settings. These technologies also played an important role during post-treatment surveillance for early detection of persistences, recurrences, and metachronous tumours.

Epidemiology and clinical features of gastroenteritis in hospitalised children: prospective survey during a 2-year period in a Parisian hospital, France.

Rotavirus is recognised as the most important agent of severe acute gastroenteritis (AGE) in young children. In a 2-year prospective survey, we investigated the epidemiology and clinical features of the viral and bacterial pathogens in children hospitalised for AGE. The study was performed in a Parisian teaching hospital from November 2001 to May 2004. Clinical data were prospectively collected to assess the gastroenteritis severity (20-point Vesikari severity score, the need for intravenous rehydration, duration of hospitalisation). Stools were systematically tested for group A rotavirus, norovirus, astrovirus and adenovirus 40/41, sapovirus and Aichi virus and enteropathogenic bacteria. A total of 457 children (mean age 15.9 months) were enrolled. Viruses were detected in 305 cases (66.7%) and bacteria in 31 cases (6.8%). Rotaviruses were the most frequent pathogen (48.8%), followed by noroviruses (8.3%) and adenoviruses, astroviruses, Aichi viruses and sapoviruses in 3.5%, 1.5%, 0.9% and 0.4%, respectively. Cases of rotavirus gastroenteritis were significantly more severe than those of norovirus with respect to the Vesikari score, duration of hospitalisation and the need for intravenous rehydration. Rotaviruses were the most frequent and most severe cause in children hospitalised for AGE, and noroviruses also account for a large number of cases in this population.

Endoscopic treatment of Zenker's diverticulum by carbon dioxide laser.

The management of Zenker's diverticulum remains controversial, as both external and endoscopic procedures are potentially associated with post-operative complications and risks. The endoscopic approach is based on cricopharyngeal myotomy or myectomy by laser, cautery or stapling. A retrospective chart review has been made from December 1994 to April 2009 of all patients with Zenker's diverticulum treated by endoscopic cricopharyngeal myectomy using CO(2) laser at the Department of Otorhinolaryngology - Head and Neck Surgery of the University of Brescia, Italy. Of the 28 patients (19 males, 9 females; mean age, 64 years; range, 46-95) scheduled for the trans-oral procedure, 27 (96%) were endoscopically treated. Diagnosis was based on the patient's history, flexible fiberoptic examination of the larynx, hypopharynx, and oesophagus, and videofluoroscopy with barium. Exclusion criteria included severe medical co-morbidities, impossibility to endoscopically expose the diverticulum, and small (< 2 cm) or large (> 6 cm) diverticula. The endoscopic procedure was performed using a CO(2) laser to section the cricopharyngeal muscle and remove the posterior part in order to obtain partial myectomy. Major complications occurred in 2 (7%) patients. No cases of recurrent nerve paralysis, pharyngo-cutaneous fistula, neck emphysema, post-operative bleeding, mediastinitis or aspiration pneumonia were observed in the present series. A swallow survey was obtained by telephone: 85% of patients reported improved swallowing (without symptoms in 11 and with moderate dysphagia in 7). Based on the present series, endoscopic CO(2) laser cricopharyngeal myectomy for Zenker's diverticulum can be considered an effective and safe procedure, with reduced hospitalization time and complication rate.

Narrow band imaging and high definition television in evaluation of oral and oropharyngeal squamous cell cancer: a prospective study.

Narrow band imaging (NBI) is an optical technique in which filtered light enhances superficial neoplasms based on their neoangiogenic pattern. The accuracy of NBI can be augmented by combining it with high definition television (HDTV). The aim of this study was to prospectively assess the diagnostic value of NBI in combination with HDTV in evaluation of oral (O) and oropharyngeal (OP) squamous cell carcinoma (SCC). Between April 2007 and December 2009, we analyzed 96 patients who were divided into 2 groups: Group A included 35 patients previously biopsied and diagnosed with OSCC or OPSCC and subjected to pre- and intraoperative HDTV white light (WL) and HDTV NBI endoscopy; Group B included 61 subjects already treated for OSCC or OPSCC and followed-up with HDTV WL and HDTV NBI. Fourteen of 35 (40%) patients in Group A showed adjunctive findings with NBI compared to standard WL. All of these findings were histologically confirmed. Twelve of 61 (20%) patients in Group B showed positive NBI findings, which were all confirmed by histology. The sensitivity, specificity, positive, negative predictive values, and accuracy for HDTV WL were 51%, 100%, 100%, 87%, and 68%, respectively, whilst for HDTV NBI were 96%, 100%, 100%, 93%, and 97%, respectively. Overall, 26 of 96 (27%) patients had a diagnostic advantage in applying NBI and HDTV: 6 patients received a diagnosis of recurrence and 1 of persistence after previous treatments; 5 showed a metachronous tumour; in 4 a synchronous tumour was diagnosed; 9 lesions were upstaged; in 1 patient previously diagnosed with an unknown primary by fine needle aspiration cytology on the neck, an anterior tonsillar pillar cancer was identified.

Prevalence and genetic diversity of Aichi virus strains in stool samples from community and hospitalized patients.

Aichi virus has been proposed as a causative agent of gastroenteritis. A total of 457 stool specimens from children hospitalized with acute diarrhea and 566 stool specimens from adults and children involved in 110 gastroenteritis outbreaks were screened for the presence of Aichi virus by reverse transcription-PCR (RT-PCR) amplification of the genomic region of the 3C and 3D (3CD) nonstructural proteins. Our results show a low incidence of Aichi virus in pediatric samples and the existence of mixed infections with other microbiological agents in some cases. From the outbreak survey, it appears that the presence of Aichi virus is an indicator of mixed infections causing gastroenteritis outbreaks and that it could be involved in half of the oyster-associated outbreaks. A second RT-PCR was developed to amplify a part of the VP1 gene. The phylogenetic analysis showed a good correlation between the two classifications based on 3CD and VP1 gene sequences and revealed the prevalence of genotype A in France. It also allowed us to partially describe an Aichi virus strain that could represent a new genotype, thus suggesting the existence of a certain diversity.

[Evaluation of seven immunochromatographic assays for the rapid detection of human rotaviruses in fecal specimens]. / Evaluation de sept réactifs d'immunochromatographie pour détecter les rotavirus humains dans les selles.

Seven commercially available immunochromatographic assays were tested for the rapid detection of group A rotaviruses in fecal samples compared to a enzyme immunoassay (Argene). Detection of rotaviruses in 80 ELISA positive frozen stool samples showed rates superior to 90% for three reagents (Rota Strip (Cypress Diagnostics), 98.8%; Rotascreen (Microgen), 95.0%; VIKIA Rota/Adeno (bioMérieux), 92.5%); from 82.5% to 88.8% for three others (Diarlex with centrifugation (Orion Diagnostica), 88.8%; Combo Rota/Adeno (All Diag), 87.5%; Rota/Adeno Combi Stick (bmd), 82.5%) and only 70.0% for Diarlex with filtration vial (Orion Diagnostica). The evaluation of the specificity, performed on one hundred fresh rotavirus negative stools, did not show any false positives with any assay. Analysis of the different technical features of these tests showed that they are quick and suitable for a clinical laboratory and do not require expensive equipment.

A large multi-pathogen waterborne community outbreak linked to faecal contamination of a groundwater system, France, 2000.

A large waterborne outbreak of infection that occurred during August 2000 in a local community in France was investigated initially via a rapid survey of visits to local physicians. A retrospective cohort study was then conducted on a random cluster sample of residents. Of 709 residents interviewed, 202 (28.5%) were definite cases (at least three liquid stools/day or vomiting) and 62 (8.7%) were probable cases (less than three liquid stools/day or abdominal pain). Those who had drunk tap water had a three-fold increased risk for illness (95% CI 2.4-4.0). The risk increased with the amount of water consumed (chi-square trend: p < 0.0001). Bacteriological analyses of stools were performed for 35 patients and virological analyses for 24 patients. Campylobacter coli, group A rotavirus and norovirus were detected in 31.5%, 71.0% and 21% of samples, respectively. An extensive environmental investigation concluded that a groundwater source to this community had probably been contaminated by agricultural run-off, and a failure in the chlorination system was identified. This is the first documented waterborne outbreak of infection involving human C. coli infections. A better understanding of the factors influencing campylobacter transmission between hosts is required.

Characterization of new recombinant noroviruses.

Noroviruses are important etiologic agents of acute gastroenteritis and show great genetic diversity. To characterize more fully previously detected strains that could not be assigned unequivocally to one particular genotype based on the RNA polymerase, we have sequenced a region in the capsid gene and, in some cases, in the junction between open reading frame 1 (ORF 1) and ORF 2. The results allowed us to identify several recombinant noroviruses: GGIIb viruses were detected for the first time in France in August 2000 and then spread through France and to Europe during the following winter. Here we present the characterization of three other probable GII recombinants which showed different phylogenetic positions depending on their ORF 1 and ORF 2 sequences. Analysis of the region located between ORF 1 and ORF 2 by a nucleotide identity window search showed a sudden shift in similarities. Moreover, recombination breakpoints were identified upstream and downstream of the beginning of ORF 2 by using a statistical test, thus confirming the involvement of this region in recombination. Unlike GGIIb, the three recombinants described here do not seem to have diffused widely in the community: one was found in a waterborne outbreak, and the other two were found in sporadic cases. Recombination is important for the evolution of RNA viruses and has already been described for noroviruses. Our results suggest that recombination is not a rare phenomenon among noroviruses, but not all these presumed recombinants that formed during RNA replication are able to spread widely.

Molecular epidemiology of caliciviruses detected in sporadic and outbreak cases of gastroenteritis in France from December 1998 to February 2004.

We compiled sequence and epidemiological data from 172 caliciviruses detected in France from December 1998 to February 2004 in sporadic and outbreak cases. The results showed a cocirculation of strains with a majority of genogroup II (GII) noroviruses. Three groups of noroviruses, not detected before in our laboratory, emerged and spread during the period: the recombinant GGIIb and Norwalk-related strains not amplified in the polymerase gene in 2000 and a new Lordsdale variant in 2002. We observed that (i) GII-4 noroviruses were predominant in nursing home and hospital outbreaks but rare in oyster- and water-related outbreaks despite continuous circulation in the population; (ii) at the opposite, genogroup I strains were detected in the majority of environmental outbreaks; (iii) several strains were frequently found in oyster- and water-linked outbreaks (up to seven), whereas one single strain was detected when transmission was from person to person; and (iv) whereas GII noroviruses were predominant in sporadic cases where patients were under 15 years of age, GI strains were more frequent in outbreaks occurring in this age group. Finally, from a methodology point of view, this compilation shows that detection and characterization in the polymerase gene are not adequate in a significant number of cases and should be completed by amplification and sequencing in the capsid gene.

Development and evaluation of a new commercial test allowing the simultaneous detection of noroviruses and sapoviruses by reverse transcription-PCR and microplate hybridization.

This work describes the design and initial evaluation of a commercial test allowing the detection of noroviruses and sapoviruses by reverse transcription-PCR (RT-PCR) in a single tube followed by microplate hybridization, as well as the detection of PCR inhibitors. The test was shown to be broadly reactive (except for Melksham-like strains), sensitive, and specific and thus should be useful for calicivirus detection in clinical practice.

Quantitative and kinetic evolution of wound healing through image analysis.

To define a healing function based on parameters measured on digitized images of wounds, and to use it to compare the rate of healing of two skin graft donor sites, one treated with petrolatum gauze (Pg) and the other with a topical preparation containing alginates (A). Digital photographs of donor sites (depth 0.6 mm) taken every two days between day 6 and day 12 were analyzed blind using the same algorithm, following changes in color and homogeneity. Analysis of variance was used to identify those parameters that changed during healing. The healing function was constructed using measurements made in six patients (group 1) randomly chosen from ten requiring skin grafts, and was applied and validated using data from the remaining four patients (group 2). The results given by this healing function were compared with those provided by principal component analysis. The most significant healing parameters were those measuring wound homogeneity, and our healing function reflects how these change with time. The time-dependent curves of the function calculated for groups 1 and 2 matched well enough to be considered as being derived from the same set of measurements. The results given by this healing function explained, by analogy, the meaning of the first principal component of principal component analysis. From day 6 to day 12, the healing function followed the same time-course for the Pg and A treatments, but healing was achieved significantly earlier (4 days, p < 0.03) with A. This suggests that the effect of A on wound healing is achieved in the first six days, before the visual changes from epidermalization are analyzable.

G and P genotyping of rotavirus strains circulating in france over a three-year period: detection of G9 and P[6] strains at low frequencies. The AZAY Group.

Rotavirus G and P types from 716 children with acute diarrhea in Dijon from 1995 to 1998 and throughout France during the winter of 1997-1998 were analyzed by reverse transcription-PCR. P[8],G1 predominated, followed by P[8],G4, which emerged during the last winter. G9 and P[6] strains were detected at low frequencies.

Prevalence of group A rotavirus, human calicivirus, astrovirus, and adenovirus type 40 and 41 infections among children with acute gastroenteritis in Dijon, France.

Group A rotaviruses, human caliciviruses, astroviruses, and adenovirus types 40 and 41 were detected by enzyme immunoassay or reverse transcription-PCR in 61, 14, 6, and 3% of stool specimens from 414 children consulting for gastroenteritis between 1995 and 1998. These data highlight the importance of caliciviruses in infantile gastroenteritis. Among these, Norwalk-like viruses belonging to genogroup II were predominant.

Assessment of healing kinetics through true color image processing.

A quantitative method of skin healing assessment using true color image processing is presented. The method was developed during a clinical trial using healthy volunteers, the goal of which was to study a drug for accelerating healing. Photographic images of the skin were sequentially acquired between day 1 and day 12 after pure painless epidermal wounds. The images were digitized in controlled conditions using a color video camera connected to a computer system. A color threshold based segmentation was developed to provide an operator-independent delineation of the wound. Two healing indexes were built measuring, the wound area and the wound color. The method was implemented in a software system allowing a fully automated determination of the healing indexes. The method provides a new quantitative global assessment of healing kinetics. It is noninvasive and well suited for multicentric clinical trials.

Is rhodamine 123 a photosensitizer?

Because of conflicting reports on the photoxicity of rhodamine 123 (Rh 123), we have undertaken a study of Rh 123 photosensitization in several in vitro systems. First, Rh 123 is not a photodynamic agent and does not react with singlet oxygen. Second, when bound to cytochrome c (Cyt c), Rh 123 photosensitizes ferro Cyt c but not ferri Cyt c degradation by an oxygen-independent process. When delivered to skin fibroblasts where it specifically stains mitochondria, Rh 123 photosensitizes membrane damage. These results are consistent with the hypothesis that Rh 123 is a phototoxic stain. The lack of photosensitivity of Rh 123-stained mitochondria in some cell lines might therefore be due to specific structural features.

[Comparative topography of the catalytic sites of chromosomal dihydrofolate reductases and that of the enzyme encoded in R 67 plasmid from E. coli]. / Topographie comparée des sites catalytiques des dihydrofolates réductases d'origine chromosomique et de l'enzyme codé par le plasmide R 67 de E. coli.

The trimethoprim-resistant dihydrofolate reductase specified in E. coli by plasmid R 67, when compared, with other enzymes catalyzing the same reaction, to have a dissimilar primary, secondary, tertiary and quaternary structure. In regard to the tertiary structure, we show here that the pteridine binding site, in the plasmid-encoded enzyme, has a geometrical similarity with that of other chromosomal specified reductases.