The TeloDIAG: how telomeric parameters can help in glioma rapid diagnosis and liquid biopsy approaches.

BACKGROUND: In glioma, TERT promoter mutation and loss of ATRX (ATRX loss) are associated with reactivation of telomerase or alternative lengthening of telomeres (ALT), respectively, i.e. the two telomere maintenance mechanisms (TMM). Strangely, 25% of gliomas have been reported to display neither or both of these alterations. MATERIALS AND METHODS: The C-circle (CC) assay was adapted to tumor (formalin-fixed paraffin-embedded and frozen) and blood samples to investigate the TMM. RESULTS: We constructed a CC-based algorithm able to identify the TMM and reported a sensitivity of 100% and a specificity of 97.3% (n = 284 gliomas). By combining the TMM, the mutational status of the isocitrate dehydrogenase 1/2 (IDH) gene (IDHmt), and the histological grading, we propose a new classification tool: TeloDIAG. This classification defined five subtypes: tOD, tLGA, tGBM_IDHmt, tGBM, and tAIV, corresponding to oligodendroglioma, IDHmt low-grade astrocytoma, IDHmt glioblastoma, and IDHwt glioblastoma (GBM), respectively; the last class gathers ALT+ IDHwt gliomas that tend to be related to longer survival (21.2 months) than tGBM (16.5 months). The TeloDIAG was 99% concordant with the World Health Organization classification (n = 312), and further modified the classification of 55 of 144 (38%) gliomas with atypical molecular characteristics. As an example, 14 of 69 (20%) of TERTwt, ATRXwt, and IDHwt GBM were actually tAIV. Outstandingly, CC in blood sampled from IDHmt astrocytoma patients was detected with a sensitivity of 56% and a specificity of 97% (n = 206 gliomas and 30 healthy donors). CONCLUSION: The TeloDIAG is a new, simple, and effective tool helping in glioma diagnosis and a promising option for liquid biopsy.

Characteristics of IDH-mutant gliomas with non-canonical IDH mutation.

BACKGROUND: Approximately 10% of IDH-mutant gliomas harbour non-canonical IDH mutations (non-p.R132H IDH1 and IDH2 mutations). OBJECTIVE: The aim of this study was to analyse the characteristics of non-canonical IDH-mutant gliomas. MATERIALS AND METHODS: We retrospectively analysed the characteristics of 166 patients with non-canonical IDH mutant gliomas and compared them to those of 155 consecutive patients with IDH1 p.R132H mutant gliomas. RESULTS: The median age at diagnosis was 38 years in patients with non-canonical IDH mutant gliomas and 43 years in glioma patients with IDH1 p.R132H-mutant tumours. Family history of cancer was more frequent among glioma patients harbouring non-canonical IDH mutations than in patients with IDH1 p.R132H mutations (22.2% vs 5.1%; P < 0.05). Tumours were predominantly localised in the frontal lobe regardless of the type of IDH mutation. Compared to IDH1 p.R132H-mutant gliomas, tumours with non-canonical IDH mutations were more frequently found in the infratentorial region (5.5% vs 0%; P < 0.05) and were often multicentric (4.8% vs 0.9%; P < 0.05). Compared to IDH1 P.R132H-mutant gliomas, tumours with non-canonical IDH1 mutations were more frequently astrocytomas (65.6% vs 43%, P < 0.05), while those with IDH2 mutations were more frequently oligodendrogliomas (85% vs 48.3%; P < 0.05). The median overall survival was similar in patients with IDH1 p.R132H-mutant gliomas and patients with non-canonical IDH-mutant gliomas. CONCLUSION: Gliomas with non-canonical IDH mutations have distinct radiological and histological characteristics. The presence of such tumours seems to be associated with genetic predisposition to cancer development.

T helper 2 inflammatory markers are associated with recurrence in chronic rhinosinusitis with nasal polyps after endoscopic sinus surgery.

BACKGROUND: Despite maximum medical treatment and endoscopic sinus surgery (ESS), chronic rhinosinusitis with nasal polyps (CRSwNP) can require revision surgery. With a growing literature on the diversity of cytokine inflammation patterns in CRSwNP, an endotype-driven approach could lead to the identification of cytokine profiles that predict recurrence. METHODS: A monocentric longitudinal study was carried out until June 2019 following CRSwNP patients who underwent surgery for the first time between December 2010 and January 2012. The biomarker profiles were established on blood and nasal secretions at the time of the first surgery (Interleukin (IL)-5, IgE, IgA, eosinophilic cationic protein (ECP) and eosinophilic- derived neurotoxin (EDN)). Profiles were compared between the patients still controlled by medical treatment and the patients requiring revision surgery during the course of the follow-up period. RESULTS: Among the 48 patients initially enrolled in our study, 8 required revision surgery (16,7%). Clinical features (asthma, allergy, aspirin intolerance, active smoking) and levels of blood markers measured at the time of the first surgery were comparable between the 2 groups of patients. Levels of IL-5, IgE and ECP in nasal secretions were significantly increased in the group of patients needing revision surgery. CONCLUSIONS: Based on simple approach of nasal secretions sampling, we showed that a predominant T helper 2 proteins expression profile can be associated with recurrent CRSwNP after ESS. Initial immunoprofiling in CRSwNP disease may contribute to better predict the therapeutic response to optimal medical and surgical treatment, and help define the role of innovative targeted treatment, beside corticosteroids and ESS.

Spondylodiscitis after transoral robotic surgery: Retrospective 7-case series from the GETTEC group.

BACKGROUND: Cervical spondylodiscitis is a rare but severe complication of pharyngeal surgery. MATERIAL AND METHODS: This multicenter retrospective study reported all patients in the database of the French head and neck tumor study group (GETTEC) affected by cervical spondylodiscitis after transoral robotic surgery (TORS) for malignant pharyngeal tumor from January 2010 to January 2017. OBJECTIVES: To describe cases of post-TORS cervical spondylodiscitis, identify alarm signs, and determine optimal management of these potentially lethal complications. RESULTS: Seven patients from 6 centers were included. Carcinomas were located in the posterior pharyngeal wall. Tumor stage was T1 or T2. All patients had risk factors for spondylodiscitis. Mean time to diagnosis was 12.6days. The interval between surgery and spondylodiscitis diagnosis ranged from 20days to 4.5months, for a mean 2.1months. The most common symptom was neck pain (87%). Infections were polymicrobial; micro-organisms were isolated in 5 cases and managed by intravenous antibiotics, associated to medullary decompression surgery in 3 cases. Follow-up found favorable progression in 4 cases, and 3 deaths (mortality, 43%). CONCLUSION: This French multicenter study found elevated mortality in post-TORS spondylodiscitis, even in case of limited resection. Surgeons must be aware of this complication and alerted by persistent neck pain, fever, asthenia, impaired or delayed posterior pharyngeal wall wound healing or elevation of inflammatory markers. MRI is the most effective diagnostic radiological examination.

Single-cell gene expression signatures reveal melanoma cell heterogeneity.

It is well established that tumours are not homogenous, but comprise cells with differing invasive, proliferative and tumour-initiating potential. A major challenge in cancer research is therefore to develop methods to characterize cell heterogeneity. In melanoma, proliferative and invasive cells are characterized by distinct gene expression profiles and accumulating evidence suggests that cells can alternate between these states through a process called phenotype switching. We have used microfluidic technology to isolate single melanoma cells grown in vitro as monolayers or melanospheres or in vivo as xenografted tumours and analyse the expression profiles of 114 genes that discriminate the proliferative and invasive states by quantitative PCR. Single-cell analysis accurately recapitulates the specific gene expression programmes of melanoma cell lines and defines subpopulations with distinct expression profiles. Cell heterogeneity is augmented when cells are grown as spheres and as xenografted tumours. Correlative analysis identifies gene-regulatory networks and changes in gene expression under different growth conditions. In tumours, subpopulations of cells that express specific invasion and drug resistance markers can be identified amongst which is the pluripotency factor POUF51 (OCT4) whose expression correlates with the tumorigenic potential. We therefore show that single-cell analysis can be used to define and quantify tumour heterogeneity based on detection of cells with specific gene expression profiles.

Tau exon 2 responsive elements deregulated in myotonic dystrophy type I are proximal to exon 2 and synergistically regulated by MBNL1 and MBNL2.

The splicing of the microtubule-associated protein Tau is regulated during development and is found to be deregulated in a growing number of pathological conditions such as myotonic dystrophy type I (DM1), in which a reduced number of isoforms is expressed in the adult brain. DM1 is caused by a dynamic and unstable CTG repeat expansion in the DMPK gene, resulting in an RNA bearing long CUG repeats (n>50) that accumulates in nuclear foci and sequesters CUG-binding splicing factors of the muscle blind-like (MBNL) family, involved in the splicing of Tau pre-mRNA among others. However, the precise mechanism leading to Tau mis-splicing and the role of MBNL splicing factors in this process are poorly understood. We therefore used new Tau minigenes that we developed for this purpose to determine how MBNL1 and MBNL2 interact to regulate Tau exon 2 splicing. We demonstrate that an intronic region 250 nucleotides downstream of Tau exon 2 contains cis-regulatory splicing enhancers that are sensitive to MBNL and that bind directly to MBNL1. Both MBNL1 and MBNL2 act as enhancers of Tau exon 2 inclusion. Intriguingly, the interaction of MBNL1 and MBNL2 is required to fully reverse the mis-splicing of Tau exon 2 induced by the trans-dominant effect of long CUG repeats, similar to the DM1 condition. In conclusion, both MBNL1 and MBNL2 are involved in the regulation of Tau exon 2 splicing and the mis-splicing of Tau in DM1 is due to the combined inactivation of both.

Management of subluxated lens in young patients.

OBJECTIVES: To evaluate visual outcomes in patients treated for lens subluxation. Secondary objectives are to report best corrected visual acuity (BCVA) in LogMAR and compare the outcomes of patients managed conservatively with those treated surgically. METHODS: Retrospective comparison of BCVA in patients under 50 years-old with lens subluxation, managed conservatively or surgically. RESULTS: A total of 49 eyes of 28 patients were included. Demographic characteristics were similar in both groups. Twenty eyes were treated surgically (40.8%) compared to 29 with medical treatment (59.2%). Marfan syndrome (79.6%) was diagnosed in 39 eyes. LogMAR BCVA post intervention was 0.35±0.31 for medical treatment and 0.39±0.32 for the surgical group, with no significant differences (P=.63). Improvements in LogMAR lines were 2.7±4.2 and 4.11±4.2 (P=.35), respectively. Two eyes in the surgery group developed ocular hypertension (0.04%), none with retinal detachment. CONCLUSIONS: The final BCVA showed no significant differences in this group of patients. BCVA depends on the visual potential of the rehabilitated eye rather than a specific type of intervention.

Manejo del cristalino subluxado en pacientes jóvenes / Management of subluxated lens in young patients

Objetivos: Evaluar los resultados visuales en pacientes tratados por subluxación del cristalino. Los objetivos secundarios son reportar la agudeza mejor corregida (AVMC) en LogMAR y comparar los resultados de los pacientes manejados de manera conservadora con aquellos manejados quirúrgicamente. Métodos: Estudio retrospectivo comparativo de AVMC en pacientes menores de 50 años con subluxación de cristalino, manejados de manera conservadora con lentes o tratados quirúrgicamente. Resultados: Se incluyeron 49 ojos de 28 pacientes. Las características demográficas fueron similares en ambos grupos; 20 ojos fueron tratados quirúrgicamente (40,8%) versus 29 con tratamiento médico (59,2%); 39 ojos con diagnóstico de síndrome de Marfán (79,6%). La AVMC LogMAR postintervención de 0,35±0,31 para el tratamiento médico y 0,39±0,32 para el grupo de manejo quirúrgico, sin encontrarse diferencias significativas (p=0,63). Las mejorías en líneas LogMAR fueron, respectivamente, 2,7±4,2 y 4,11±4,2 (p=0,35). Dos ojos del grupo cirugía evolucionaron con hipertensión ocular (0,04%), ninguno con DR. Conclusiones: La AVMC final no presentó diferencias significativas en este grupo de pacientes, dependiendo del potencial visual del ojo rehabilitado más que de un tipo específico de intervención(AU)

Objectives: To evaluate visual outcomes in patients treated for lens subluxation. Secondary objectives are to report best corrected visual acuity (BCVA) in LogMAR and compare the outcomes of patients managed conservatively with those treated surgically. Methods: Retrospective comparison of BCVA in patients under 50 years-old with lens subluxation, managed conservatively or surgically. Results: A total of 49 eyes of 28 patients were included. Demographic characteristics were similar in both groups. Twenty eyes were treated surgically (40.8%) compared to 29 with medical treatment (59.2%). Marfan syndrome (79.6%) was diagnosed in 39 eyes. LogMAR BCVA post intervention was 0.35±0.31 for medical treatment and 0.39±0.32 for the surgical group, with no significant differences (P=.63). Improvements in LogMAR lines were 2.7±4.2 and 4.11±4.2 (P=0.35), respectively. Two eyes in the surgery group developed ocular hypertension (0.04%), none with retinal detachment. Conclusions: The final BCVA showed no significant differences in this group of patients. BCVA depends on the visual potential of the rehabilitated eye rather than a specific type of intervention(AU)

Mis-splicing of Tau exon 10 in myotonic dystrophy type 1 is reproduced by overexpression of CELF2 but not by MBNL1 silencing.

Tau is the proteinaceous component of intraneuronal aggregates common to neurodegenerative diseases called Tauopathies, including myotonic dystrophy type 1. In myotonic dystrophy type 1, the presence of microtubule-associated protein Tau aggregates is associated with a mis-splicing of Tau. A toxic gain-of-function at the ribonucleic acid level is a major etiological factor responsible for the mis-splicing of several transcripts in myotonic dystrophy type 1. These are probably the consequence of a loss of muscleblind-like 1 (MBNL1) function or gain of CUGBP1 and ETR3-like factor 1 (CELF1) splicing function. Whether these two dysfunctions occur together or separately and whether all mis-splicing events in myotonic dystrophy type 1 brain result from one or both of these dysfunctions remains unknown. Here, we analyzed the splicing of Tau exons 2 and 10 in the brain of myotonic dystrophy type 1 patients. Two myotonic dystrophy type 1 patients showed a mis-splicing of exon 10 whereas exon 2-inclusion was reduced in all myotonic dystrophy type 1 patients. In order to determine the potential factors responsible for exon 10 mis-splicing, we studied the effect of the splicing factors muscleblind-like 1 (MBNL1), CUGBP1 and ETR3-like factor 1 (CELF1), CUGBP1 and ETR3-like factor 2 (CELF2), and CUGBP1 and ETR3-like factor 4 (CELF4) or a dominant-negative CUGBP1 and ETR-3 like factor (CELF) factor on Tau exon 10 splicing by ectopic expression or siRNA. Interestingly, the inclusion of Tau exon 10 is reduced by CUGBP1 and ETR3-like factor 2 (CELF2) whereas it is insensitive to the loss-of-function of muscleblind-like 1 (MBNL1), CUGBP1 and ETR3-like factor 1 (CELF1) gain-of-function, or a dominant-negative of CUGBP1 and ETR-3 like factor (CELF) factor. Moreover, we observed an increased expression of CUGBP1 and ETR3-like factor 2 (CELF2) only in the brain of myotonic dystrophy type 1 patients with a mis-splicing of exon 10. Taken together, our results indicate the occurrence of a mis-splicing event in myotonic dystrophy type 1 that is induced neither by a loss of muscleblind-like 1 (MBNL1) function nor by a gain of CUGBP1 and ETR3-like factor 1 (CELF1) function but is rather associated to CUGBP1 and ETR3-like factor 2 (CELF2) gain-of-function.

All the 1p19q codeleted gliomas are mutated on IDH1 or IDH2.

BACKGROUND: Recently, the gene encoding the human cytosolic NADPH-dependent isocitrate dehydrogenase (IDH1) was reported frequently mutated in gliomas. Rare mutations were also found in the sequence of the mitochondrial isoform IDH2. METHODS: In a series of 764 gliomas genome-wide characterized, we determined the presence of mutations in the sequences of both IDH1 and IDH2 genes by direct sequencing. RESULTS: We found that all tumors with complete 1p19q codeletion (n = 128) were mutated in the IDH1 (118) or IDH2 (10) gene. This 100% mutation rate contrasted strikingly with other gliomas exhibiting either variable 1p and 19q alterations (n = 159, IDH1/IDH2 mutation rate of 33%) or no 1p19q alteration (n = 477, IDH1/IDH2 mutation rate 32%). Our data also confirm the prognostic impact of IDH1/IDH2 mutation in gliomas whatever grade considered: patients harboring mutations of IDH1/IDH2 have an improved median overall survival. Moreover, in WHO grade II and III gliomas, 3 groups with significantly different outcomes were identified according to their 1p19q and IDH1/IDH2 statuses. Tumors carrying both alterations had longer overall survival than their nonmutated counterpart. CONCLUSIONS: This exclusive association suggests a new mechanism of tumorigenesis. Perhaps the IDH1/IDH2 mutation is a prerequisite for the occurrence of the t(1;19) translocation, or it is required for the 1p19q codeleted cells to acquire a tumor phenotype.

Bevacizumab and irinotecan for recurrent oligodendroglial tumors.

BACKGROUND: Treatment with a regimen of bevacizumab-irinotecan has been shown to be effective in recurrent grade 3 and 4 gliomas, but the effect of this regimen against recurrent oligodendroglial tumors has not been specifically studied. METHODS: The bevacizumab-irinotecan regimen was retrospectively evaluated in a consecutive series of 25 patients with recurrent oligodendroglial tumors. All patients had not responded to previous treatment with radiation therapy and at least one line of temozolomide chemotherapy. Bevacizumab (10 mg/kg) and irinotecan (125 or 340 mg/m(2) according to the antiepileptic regimen) were administered every 14 days. Response was measured clinically and on monthly MRI. RESULTS: The objective response rate was 72% (20% complete response, 52% partial response). After a median follow up of 202 days, the median progression-free survival was 140 days (95% confidence interval [CI] 116-infinity), and overall survival had not been reached. The 6-month progression-free survival was 42% (95% CI 26%-67%). Among the 17 patients in whom the status of the main molecular alterations of gliomas could be evaluated (search for deletions of chromosomes 1p, 19q, 9p, and 10q and amplification of epidermal growth factor receptor, mouse double-minute gene, and cyclin-dependent kinase 4 gene), no relation could be found between the response rate and the type of genetic change (including 1p-19q codeletion). The profile of tolerance was fair, with treatment discontinuation in 20% of patients. Intratumoral hemorrhages occurred in 6 patients (24%), but the treatment had to be discontinued because of symptomatic bleeding in only 1 patient (4%). CONCLUSIONS: This regimen is effective in recurrent oligodendrogliomas, and the overall tolerance is acceptable.

Polymorphism in Sp1 recognition site of the EGF receptor gene promoter and risk of glioblastoma.

We investigated two polymorphisms of the epidermal growth factor receptor promoter as potential risk factors and prognostic markers for glioblastoma. The -216T allele (which results in a 30% higher activity) was more frequent in the patients compared with the control population (224/376 = 59.6% vs 165/352 = 46.8%; p = 0.0006) corresponding to an odd ratio of 1.67 (1.24; 2.25). A modest difference in median survival was also associated with the TT genotype.

[Co-sleeping and sudden infant death syndrome]. / Cosleeping et mort subite du nourrisson.

Bedsharing has recently become a controversial subject. Some authors, mainly from North America, assign to bedsharing a positive effect on the efficacy and duration of breast-feeding. Moreover, it would protect against sudden infant death syndrome (SIDS). Conversely, other studies consider bedsharing as an additional risk factor for SIDS. From the literature data, there is some evidence for an increased risk of SIDS when bedsharing is associated with maternal smoking and alcohol consumption. Bedsharing cannot be recommended as an absolutely safe practice. Breast-feeding mothers should be aware of these potential hazards.

[Evaluation of retinal involvement in type 2 diabetics with microalbuminuria]. / Evaluación del compromiso retinal en diabéticos tipo 2 microalbuminúricos.

BACKGROUND: In patients with type 2 diabetes, the presence of microalbuminuria, reflecting a widespread vascular damage, can be a marker of nephropathy, retinophaty and cardiovascular diseases. AIM: To study the relationship between microalbuminuria and the frequency, severity and outcome of retinopathy in patients with type 2 diabetes mellitus. PATIENTS AND METHODS: One hundred patients with type 2 diabetes were subjected to a clinical examination, serial monitoring of blood pressure and quarterly measurement of microalbuminuria by RIA. Annually, a fundoscopy, a color photography of the posterior pole and retinal angiofluorescence were performed. Retinopathy was classified as basal (mild to moderate), preproliferative and proliferative. Sixty-four normoalbuminuric patients (urinary albumin of less than 30 mg/24 h) were included in group 1 and 36 patients with a urinary albumin over 30 mg/24 h in group 2. Fifty seven patients with normal blood pressure were randomly treated with enalapril or placebo and those with hypertension received enalapril or acebutolol to normalize blood pressure. RESULTS: Sixty one percent of group 1 patients and 41% of group 2 patients has retinopathy (p < 0.05). The retinal lesions were proliferative in 41% of group 2 patients and in 8% of group 1 patient (p < 0.05). Retinopathy was present in 67% of hypertensive patients of group 2 and in 41% of hypertensive patients of group 1. An unfavorable evolution of retinopathy was observed in 22% of group 2 patients and in 5% of group 1 patient (p < 0.05). CONCLUSIONS: In type 2 diabetic patients, the presence of microalbuminuria is a prediction of a higher frequency, severity and dismal evolution of diabetic retinopathy (Rev Méd Chile 2000; 128: 1085-92).

[Prematurity and sudden infant death syndrome. Polysomnography in question]. / Prématurité et syndrome de mort subite du nourrisson. La polysomnographie en question.

Systematic recording of cardiorespirographic events has been recommended by some authors in premature and/or very low birth weight infants before or shortly after hospital's discharge. Their objective is the recognition of babies at risk of sudden infant death syndrome (SIDS) and prevention by home monitoring. After an extensive review of the recent literature, prematurity itself does not appear as a risk factor of SIDS. Late apneas are common, but their prognostic significance remains uncertain. Although it is clear that bronchopulmonary dysplasia carries a greater risk of acute life threatening events and infantile death, their prevention mainly relies upon an adequate oxygen supplementation. As a consequence no more than the general infant population, premature infants require neither polysomnographic recording nor home monitoring.

[Gastroesophageal reflux and esophageal motility disorders in infants with vagal hyperreflectivity presenting severe syncope]. / Reflux gastroesophagien et troubles de la motricité oesophagienne chez des nourrissons porteurs d'une hyper-réflectivité vagale ayant présenté un malaise grave.

AIM: The aim of the study was to evaluate the prevalence of gastroesophageal reflux and abnormalities of esophageal motility in a population of neonates referred for apparently life threatening event (ALTE) and presenting vagal hyperreflectivity. POPULATION AND METHODS: The study included 17 infants, who were examined after an ALTE. They were admitted at a mean age of 11.7 weeks (range 1-40 weeks). Vagal hyperreflectivity was confirmed in each infant by oculocardiac reflex. Before treatment, 24-hour intraesophageal pH-monitoring and esophageal manometry were performed. RESULTS: pH-monitoring and esophageal manometry were both normal in only two patients. pH-monitoring showed pathological reflux (% of time with pH < 4 more than 4.8%) in 10/17 (59%) patients. Manometric studies showed esophageal dysmotility in 12/17 (71%) of patients. Hypertensive lower sphincter was noted in 11/17 (65%) infants. Patients with normal manometry were older than patients presenting with esophageal dysmotility (P < 0.05). CONCLUSION: This study shows a high frequency of gastroesophageal reflux and dysmotility in infants with vagal hyperreflectivity. Hypertensive lower esophageal sphincter as well as vagal hyperreflectivity may correspond to dysmaturity of autonomous nervous system and facilitate the occurrence of ALTE.

Plasma exchange in Rasmussen's encephalitis.

The authors observed a 4-year-old girl who has Rasmussen's encephalitis. She started with frequent localized and generalized seizures. Standard antiepileptic treatment was almost ineffective. The frequency of the generalized seizures decreased, but the myoclonic jerks of the left part of the body persisted. An EEG showed partial status epilepticus. The results of the CT scan were normal. Antibodies to viruses were absent from the blood and cerebrospinal fluid. An MR scan showed a T2-weighted hypersignal zone in the right frontal region. Intravenous bolus injections of corticosteroids and drips of immunoglobulins were inefficient, and we started plasma exchanges which have continued for 9 months. The clinical state stabilized, and the images on the MR scan improved, but the results of the EEG did not improve. The authors discuss the effect of the plasma exchange, the use of which is questionable in this disease.