Whole genome sequencing in ROHHAD trios proved inconclusive: what's beyond?

Rapid-onset Obesity with Hypothalamic dysfunction, Hypoventilation and Autonomic Dysregulation (ROHHAD) is a rare, life-threatening, pediatric disorder of unknown etiology, whose diagnosis is made difficult by poor knowledge of clinical manifestation, and lack of any confirmatory tests. Children with ROHHAD usually present with rapid onset weight gain which may be followed, over months or years, by hypothalamic dysfunction, hypoventilation, autonomic dysfunction, including impaired bowel motility, and tumors of neural crest origin. Despite the lack of evidence of inheritance in ROHHAD, several studies have been conducted in recent years that have explored possible genetic origins, with unsuccessful results. In order to broaden the search for possible genetic risk factors, an attempt was made to analyse the non-coding variants in two trios (proband with parents), recruited in the Gaslini Children's Hospital in Genoa (Italy). Both patients were females, with a typical history of ROHHAD. Gene variants (single nucleotide variants, short insertions/deletions, splice variants or in tandem expansion of homopolymeric tracts) or altered genomic regions (copy number variations or structural variants) shared between the two probands were searched. Currently, we have not found any potentially pathogenic changes, consistent with the ROHHAD clinical phenotype, and involving genes, regions or pathways shared between the two trios. To definitively rule out the genetic etiology, third-generation sequencing technologies (e.g., long-reads sequencing, optical mapping) should be applied, as well as other pathways, including those associated with immunological and autoimmune disorders, should be explored, making use not only of genomics but also of different -omic datasets.

Análisis factorial confirmatorio del cuestionario sobre hábitos saludables en adolescentes ENHASA / Confirmatory factor analysis of the habits questionnaire healthy adolescents ENHASA

Introducción. Las cifras de sobrepeso y obesidad durante la infancia y la adolescencia están creciendo a un ritmoalarmante. Además, se han descrito diferencias en los estilos de vida entre chicos y chicas. Detectar a los individuoscon riesgo de desarrollar exceso de peso es imprescindiblepara un diagnóstico precoz y una intervención tempranaque evite la aparición de comorbilidades. Para ello es devital importancia disponer de instrumentos de cribado decalidad, debidamente analizados.Material y métodos. Se aplicó el cuestionario de HábitosSaludables en Adolescentes (ENHASA) a una muestra deestudiantes de primer curso de educación secundaria obligatoria (ESO) de la Región de Murcia. Se obtuvo además lasomatometría y se clasificaron según IMC. Se realizó análisisfactorial confirmatorio (AFC) según género para valorar laspropiedades psicométricas de la encuesta.Resultados. La muestra final fue de 416 alumnos, de losque el 40,6% presentó exceso de peso; no habiendo diferencias entre sexos. El 28,2% del total reflejó hábitos mejorables; obteniendo las peores puntuaciones en las dimensionesactividad física y uso de nuevas tecnologías. El KMO fuede 0,729 en chicos y de 0,725 en chicas; y la varianza totalexplicada fue del 61,57% y 62,92%, respectivamente. El AFCmostró concordancia estructural con respecto al análisis factorial exploratorio previo en ambos sexos. Conclusiones. Dos de cada 5 adolescentes de la muestra presentó exceso de peso. Se confirma que ENHASA esun cuestionario válido y fiable que puede utilizarse comométodo de cribado en la práctica clínica. (AU)

Introduction. The numbers of overweight and obesity during childhood and adolescence are growing atan alarming rate. In addition, differences in the lifestylesbetween boys and girls have been described. Detectingindividuals at risk of developing excess weight is essentialto carry out an early diagnosis and early intervention, andthus avoid the appearance of comorbidity. For this, it isvitally important to have quality screening instruments,duly analyzed.Methods. We applied the Healthy Habits in Adolescents(ENHASA) questionnaire to a sample of students in thefirst year of secondary education in the Region of Murcia.Somatometry was also obtained and classified accordingto BMI. Confirmatory factor analysis (CFA) was performedaccording to gender to assess the psychometric propertiesof the survey. Results. The final sample was 416 students, of which40.6% were overweight or obesity; there being no differences between the sexes. 28.2% of the total reflected habitsthat could be improved; obtaining the worst scores in thedimensions of physical activity and use of new technologies.The KMO was 0.729 in boys and 0.725 in girls; and the totalexplained variance was 61.57% and 62.92% respectively. TheCFA showed structural agreement with respect to the previous exploratory factor analysis in both sexes.Conclusion. 2 out of 5 adolescents in the sample wereexcess of weight. ENHASA is confirmed to be a valid andreliable questionnaire that can be used as a screening methodin clinical practice. (AU)

Peripheral inflammation exacerbates α-synuclein toxicity and neuropathology in Parkinson's models.

AIMS: Parkinson's disease and related disorders are devastating neurodegenerative pathologies. Since α-synuclein was identified as a main component of Lewy bodies and neurites, efforts have been made to clarify the pathogenic mechanisms of α-synuclein's detrimental effects. α-synuclein oligomers are the most harmful species and may recruit and activate glial cells. Inflammation is emerging as a bridge between genetic susceptibility and environmental factors co-fostering Parkinson's disease. However, direct evidence linking inflammation to the harmful activities of α-synuclein oligomers or to the Parkinson's disease behavioural phenotype is lacking. METHODS: To clarify whether neuroinflammation influences Parkinson's disease pathogenesis, we developed: (i) a 'double-hit' approach in C57BL/6 naive mice where peripherally administered lipopolysaccharides were followed by intracerebroventricular injection of an inactive oligomer dose; (ii) a transgenic 'double-hit' model where lipopolysaccharides were given to A53T α-synuclein transgenic Parkinson's disease mice. RESULTS: Lipopolysaccharides induced a long-lasting neuroinflammatory response which facilitated the detrimental cognitive activities of oligomers. LPS-activated microglia and astrocytes responded differently to the oligomers with microglia activating further and acquiring a pro-inflammatory M1 phenotype, while astrocytes atrophied. In the transgenic 'double-hit' A53T mouse model, lipopolysaccharides aggravated cognitive deficits and increased microgliosis. Again, astrocytes responded differently to the double challenge. These findings indicate that peripherally induced neuroinflammation potentiates the α-synuclein oligomer's actions and aggravates cognitive deficits in A53T mice. CONCLUSIONS: The fine management of both peripheral and central inflammation may offer a promising therapeutic approach to prevent or slow down some behavioural aspects in α-synucleinopathies.

Somatic gene editing ameliorates skeletal and cardiac muscle failure in pig and human models of Duchenne muscular dystrophy.

Frameshift mutations in the DMD gene, encoding dystrophin, cause Duchenne muscular dystrophy (DMD), leading to terminal muscle and heart failure in patients. Somatic gene editing by sequence-specific nucleases offers new options for restoring the DMD reading frame, resulting in expression of a shortened but largely functional dystrophin protein. Here, we validated this approach in a pig model of DMD lacking exon 52 of DMD (DMDΔ52), as well as in a corresponding patient-derived induced pluripotent stem cell model. In DMDΔ52 pigs1, intramuscular injection of adeno-associated viral vectors of serotype 9 carrying an intein-split Cas9 (ref. 2) and a pair of guide RNAs targeting sequences flanking exon 51 (AAV9-Cas9-gE51) induced expression of a shortened dystrophin (DMDΔ51-52) and improved skeletal muscle function. Moreover, systemic application of AAV9-Cas9-gE51 led to widespread dystrophin expression in muscle, including diaphragm and heart, prolonging survival and reducing arrhythmogenic vulnerability. Similarly, in induced pluripotent stem cell-derived myoblasts and cardiomyocytes of a patient lacking DMDΔ52, AAV6-Cas9-g51-mediated excision of exon 51 restored dystrophin expression and amelioreate skeletal myotube formation as well as abnormal cardiomyocyte Ca2+ handling and arrhythmogenic susceptibility. The ability of Cas9-mediated exon excision to improve DMD pathology in these translational models paves the way for new treatment approaches in patients with this devastating disease.

Multiparametric MR imaging to assess response following neoadjuvant systemic treatment in various breast cancer subtypes: Comparison between different definitions of pathologic complete response.

OBJECTIVES: To validate the performance of multiparametric magnetic resonance (MR) imaging to assess pathologic response to neoadjuvant systemic therapy (NST) in various breast cancer subtypes considering two definitions of pCR: absence of any residual invasive cancer or DCIS (ypT0) and absence of invasive tumour cells (ypT0/is). METHODS: Institutional review board-approved retrospective study, with waiver of the need to obtain informed consent. From January 2015 to June 2017, 81 women with 82 breast cancers undergoing NST were included. Eighteen lesions (22%) were immunohistochemically HER2-positive, 12 (15%) triple negative (TN), 42 (51%) luminal B-like and 10 (12%) luminal B-like/HER2-positive. Breast MR imaging was performed before and after NST. A comparative analysis considering pCR as ypT0 and ypT0/is was carried out. Performance of univariate and multivariate models to potentially predict pathologic response were evaluated. RESULTS: ypT0 was attained in 23% (19/82) of cases and ypT0/is in 33% (27/82) of cases. In both scenarios, HER2-positive subtype achieved the best response, 53% and 48%, respectively. A significant relationship was found between late enhancement and pathologic response (p < 0.001) regardless of pCR definition. In the ypT0 scenario, mean ADC ratio in the pCR subgroup was significantly higher than that in the non-pCR subgroup (p = 0.021) but no significant relationship was noted in ypT0/is. A multivariate model including MR late enhancement, ADC ratio and tumor subtype identified pathologic response with 86% and 84% accuracy when ypT0 and ypT0/is were considered, respectively. CONCLUSION: MR imaging late enhancement and ADC ratio along with breast cancer IHC subtype identify pathologic response following NST with high accuracy, achieving the highest NPV in TN and HER2-positive tumors and the highest PPV in luminal B-like subtypes, regardless of the definition of pCR as ypT0 or ypT0/is. In light of these findings and given that residual DCIS does not have an impact on survival rates, ypT0/is seems to be the preferable definition of pCR.

Ectopic teeth in the maxillary sinus: A case report and literature review.

Ectopic eruption of teeth is a rare phenomenon although there have been reports of teeth in the nasal septum, mandibular condyle, and maxillary sinus. This impaction can present itself in a variety of ways such as chronic or recurrent sinusitis, sepsis, and facial numbness and can also be asymptomatic. The aim of this study was to describe, by means of research literature and by a case report, the characteristics and occurrence of ectopic eruption in the maxillary sinus. We have analyzed and compared clinical cases of ectopic teeth in the maxillary sinus with a search on PubMed utilizing keywords such as "ectopic," "teeth," "sinus," "maxillary," and Boolean operators "or" and "and" up until 2016. Fifty-one cases were found, of which 53% were female. The age ranged between 3 and 72 years, with an average age of 28.36 years. The higher prevalence of ectopic teeth is the 3rd molars. Ten of these teeth are associated with a dentigerous cyst, 1 by an osteoma, and 2 by soft tissue. Standard treatment for an ectopic tooth is extraction, but for other patients, treatment of choice in asymptomatic ectopic tooth cases is continued observation. Ectopic teeth tend to form a cyst or tumor if not managed.

Autologous dental pulp mesenchymal stem cells for inferior third molar post-extraction socket healing: A split-mouth randomised clinical trial.

BACKGROUND: Since the discovery of adult mesenchymal stem cells extensive research has been conducted to determine their mechanisms of differentiation and effectiveness in cell therapy and regenerative medicine. MATERIAL AND METHODS: To assess the efficacy of autologous dental pulp mesenchymal stem cells delivered in a collagen matrix for post-extraction socket healing, a single-centre, double-blind, randomised, split-mouth, controlled clinical trial was performed. Both impacted mandibular third molars were extracted from 32 patients. Dental pulp was collected and dissociated; the resulting cell suspension, obtained by centrifugation, was incorporated into a resorbable collagen matrix and implanted in 32 experimental post-extraction sockets. Collagen matrices alone were implanted in 32 contralateral, control post-extraction sockets. Two neuroradiologists independently assessed the extent of bone repair at 6 months after the extractions. Computed tomography (CT, Philips Brilliance) and an advanced display platform (IntelliSpace Portal) was used to record extraction socket density, expressed as Hounsfield units (HU) and height (mm) of the distal interdental bone septum of the second molar. Measurements at 6 months post-extraction were compared with measurements obtained immediately after extraction. Data were analysed with the statistical program STATA 14. RESULTS: Two patients dropped out of the study. The final sample consisted of 22 women and 8 men (mean age, 23 years; range: 18-30 years). Clinical, radiological, and surgical characteristics of impacted third molars of the control and experimental groups were homogeneous. Measurements obtained by the two neuroradiologists showed agreement. No significant differences were found in the extent of bone repair during analyses of density (p=0.4203 neuroradiologist 1; p=0.2525 neuroradiologist 2) or interdental septum height (p=0.2280 neuroradiologist 1; p=0.4784 neuroradiologist 2). CONCLUSIONS: In our clinical trial, we were unable to demonstrate that autologous dental pulp mesenchymal stem cells reduce socket bone resorption after inferior third molar extraction.

shRNA targeting of ferritin heavy chain activates H19/miR-675 axis in K562 cells.

PURPOSE: The heavy subunit of the iron storage protein ferritin (FHC) is essential for the intracellular iron metabolism and, at the same time, it represents a central hub of iron-independent pathways, such as cell proliferation, angiogenesis, p53 regulation, chemokine signalling, stem cell expansion, miRNAs expression. In this work we have explored the ability of FHC to modulate gene expression in K562 cells, through the up-regulation of the lncRNA H19 and its cognate miR-675. MATERIALS AND METHODS: Targeted silencing of FHC was performed by lentiviral-driven shRNA strategy. FHC reconstitution was obtained by full length FHC cDNA transfection with Lipofectamine 2000. ROS amounts were determined with the redox-sensitive probe H2DCFDA. H19, miR-675, miR-107, Twist1, ID3, EPHB6, GNS, ANK1 and SMAD6 mRNA amounts were quantified by Taqman assay and qPCR analysis. RESULTS: FHC silencing in K562 cells modulates gene expression through the up-regulation of the lncRNA H19 and its cognate miR-675. Experimental findings demonstrate that the molecular mechanism underlying this phenomenon is represented by an FHC knock-down-triggered increase in reactive oxygen species (ROS) production. CONCLUSIONS: In this paper we uncover a so far not described function of the ferritin heavy subunit in the control of lncRNA pathways.

Efficacy of amoxicillin and amoxicillin/clavulanic acid in the prevention of infection and dry socket after third molar extraction. A systematic review and meta-analysis.

BACKGROUND: Prophylactic use of amoxicillin and amoxicillin/clavulanic acid, although controversial, is common in routine clinical practice in third molar surgery. MATERIAL AND METHODS: Our objective was to assess the efficacy of prophylactic amoxicillin with or without clavulanic acid in reducing the incidence of dry socket and/or infection after third molar extraction. We conducted a systematic review and meta-analysis consulting electronic databases and references in retrieved articles. We included double-blind placebo-controlled randomized clinical trials published up to June 2015 investigating the efficacy of amoxicillin with or without clavulanic acid on the incidence of the aforementioned conditions after third molar extraction. Relative risks (RRs) were estimated with a generic inverse-variance approach and a random effect model using Stata/IC 13 and Review Manager Version 5.2. Stratified analysis was performed by antibiotic type. RESULTS: We included 10 papers in the qualitative review and in the quantitative synthesis (1997 extractions: 1072 in experimental groups and 925 in controls, with 27 and 74 events of dry socket and/or infection, respectively). The overall RR was 0.350 (p<0.001; 95% CI 0.214 to 0.574). We found no evidence of heterogeneity (I2=0%, p=0.470). The number needed to treat was 18 (95% CI 13 to 29). Five studies reported adverse reactions (RR=1.188, 95% CI 0.658 to 2.146, p =0.567). The RRs were 0.563 for amoxicillin (95% CI 0.295 to 1.08, p=0.082) and 0.215 for amoxicillin/clavulanic acid (95% CI 0.117 to 0.395, p<0.001). CONCLUSIONS: Prophylactic use of amoxicillin does not significantly reduce the risk of infection and/or dry socket after third molar extraction. With amoxicillin/clavulanic acid, the risk decreases significantly. Nevertheless, considering the number needed to treat, low prevalence of infection, potential adverse reactions to antibiotics and lack of serious complications in placebo groups, the routine prescription of amoxicillin with or without clavulanic acid is not justified.

Evaluation of PCR for cutaneous leishmaniasis diagnosis and species identification using filter paper samples in Panama, Central America.

Cutaneous leishmaniasis (CL) is a major vectorborne disease in Panama. In this study, the diagnostic performance and usefulness of two DNA extraction procedures from skin scraping samples collected on FTA filter paper for subsequent PCR diagnosis of CL was evaluated. A positive CL laboratory diagnosis was based on a positive parasitological test (Giemsa-stained smears or in vitro culture) and/or positive PCR test performed from skin scrapings collected in TE buffer (PCR-TE). Of 100 patients with skin lesions suggestive of CL, 82 (82%) were confirmed as CL positive. The sensitivity was calculated for each of the PCR approaches from samples collected on filter paper. The highest sensitivity was achieved by PCR-FTA processed by Chelex 100 (PCR-Chelex) (0.94). PCR-FTA extracted using the FTA purification reagent presented a lower sensitivity (0.60). Good concordance between routine PCR-TE and PCR-Chelex was observed (percent agreement=0.88, κ index=0.65). In conclusion, use of FTA filter paper for skin scraping collection combined with PCR is a reliable and convenient method for CL diagnosis in Panama, with comparable performance to the routine PCR method and with improved sensitivity compared with those of conventional parasitological methods.

Fluorescence properties of the Na⁺/H⁺exchanger inhibitor HMA (5-(N,N-hexamethylene)amiloride) are modulated by intracellular pH.

HMA (5-(N,N-hexamethylene)amiloride), which belongs to a family of novel amiloride derivatives, is one of the most effective inhibitors of Na+/H+ exchangers, while uneffective against Na+ channels and Na+/Ca2+ exchangers. In this study, we provided evidence that HMA can act as a fluorescent probe. In fact, human retinal ARPE19 cells incubated with HMA show an intense bluish fluorescence in the cytoplasm when observed at microscope under conventional UV-excitation conditions. Interestingly, a prolonged observation under continuous exposure to excitation lightdoes not induce great changes in cells incubated with HMA for times up to about 5 min, while an unexpected rapid increase in fluorescence signal is observed in cells incubated for longer times. The latter phenomenon is particularly evident in the perinuclear region and in discrete spots in the cytoplasm. Since HMA modulates intracellular acidity, the dependence of its fluorescence properties on medium pH and response upon irradiation have been investigated in solution, at pH 5.0 and pH 7.2. The changes in both spectral shape and amplitude emission indicate a marked pH influence on HMA fluorescence properties, making HMA exploitable as a self biomarker of pH alterations in cell studies, in the absence of perturbations induced by the administration of other exogenous dyes.

Clinical presentation and pathogenesis of cold-induced autoinflammatory disease in a family with recurrence of an NLRP12 mutation.

OBJECTIVE: NLRP12 mutations have been described in patients affected with peculiar autoinflammatory symptoms. This study was undertaken to characterize NLRP12 mutations in patients with autoinflammatory syndromes, particularly a novel missense mutation, p.D294E, affecting a protein sequence crucial for ATP binding, which was identified in a Caucasian family with familial cold-induced autoinflammatory syndrome in some family members. METHODS: Fifty patients were tested for NLRP12 mutations. A Caucasian family with the p.D294E missense mutation of NLRP12 in some family members was clinically characterized. In vitro analysis of the effects of the mutation on NF-κB activity was performed in HEK 293 cells after cotransfection of the cells with a luciferase NF-κB-responsive element and mutant or wild-type (WT) NLRP12 expression plasmids. NF-κB activity was also evaluated 24 hours after stimulation with tumor necrosis factor α in monocytes from individual family members carrying the mutation. Furthermore, secretion of interleukin-1ß (IL-1ß), production of reactive oxygen species (ROS), and activation of antioxidant systems in patient and healthy donor monocytes, under resting conditions and after stimulation with pathogen-associated molecular patterns (PAMPs), were also assessed. RESULTS: In the family assessed, the p.D294E mutation segregated in association with a particular sensitivity to cold exposure (especially arthralgias and myalgia), but not always with an inflammatory phenotype (e.g., urticarial rash or fever). In vitro, the mutant protein maintained the same inhibitory activity as that shown by WT NLRP12. Consistently, NLRP12-mutated monocytes showed neither increased levels of p65-induced NF-κB activity nor higher secretion of IL-1ß. However, the kinetics of PAMP-induced IL-1ß secretion were significantly accelerated, and high production of ROS and up-regulation of antioxidant systems were demonstrated. CONCLUSION: Even with a variable range of associated manifestations, the extreme sensitivity to cold represents the main clinical hallmark in an individual carrying the p.D294E mutation of the NLRP12 gene. Although regulation of NF-κB activity is not affected in patients, redox alterations and accelerated secretion of IL-1ß are associated with this mild autoinflammatory phenotype.

[Application of the ROLL technique in a case of bilateral multifocal breast cancer]. / Aplicación de la técnica ROLL en un caso de cáncer de mama multifocal bilateral.

The increasing number of non-palpable breast lesions and the trend to perform less aggressive surgical procedures have prompted the development of radioguided surgery in breast lesions, alone (ROLL) or with sentinel node detection. We present a 55-year-old woman with four mammary lesions diagnosed by magnetic resonance. The ROLL technique allowed not only their characterisation but also the study of the lymphatic drainage of malignant lesions. Moreover, it was useful as a guide for surgical biopsy of the benign lesions. The combination of different colloid size allowed an accurate study of each lesion and avoided radical surgery. The applications of radioguided surgery are ever increasing. Individualisation of each indication will benefit a greater number of patients.

Aplicación de la técnica ROLL en un caso de cáncer de mama multifocal bilateral / Application of the ROLL technique in a case of bilateral multifocal breast cancer

El aumento de la detección de lesiones mamarias no palpables derivado de los programas de cribado ha impulsado el desarrollo de la localización radioguiada de lesiones mamarias, ya sea de forma aislada (ROLL) o junto a la detección del ganglio centinela. Presentamos el caso de una paciente con cuatro lesiones mamarias detectadas por resonancia magnética en las que la técnica ROLL permitió realizar la resección y estudio del drenaje linfático de las lesiones con confirmación de malignidad. Además, ayudó a caracterizar el resto de las lesiones sirviendo de guía en la biopsia quirúrgica. El uso de radiocoloides de diferente tamaño permitió estudiar correctamente cada lesión y evitó la realización de una cirugía más radical. Las aplicaciones de la cirugía radioguiada se amplían día a día. La individualización de las indicaciones hace posible que se beneficie un mayor número de pacientes

The increasing number of non-palpable breastlesions and the trend to perform less aggressive surgical procedures have prompted the development of radioguided surgery in breast lesions, alone (ROLL) or with sentinel node detection. We present a 55-year-old woman with four mammary lesions diagnosed by magnetic resonance. The ROLL technique allowed not only their characterisation but also the study of the lymphatic drainage of malignant lesions. Moreover, it wasuseful as a guide for surgical biopsy of the benign lesions. The combination of different colloid size allowed an accurate study of each lesion and avoided radical surgery. The applications of radioguided surgery are ever increasing. Individualisation of each indication will benefit a greater number of patients (AU)

ATM gene expression is associated with differentiation and angiogenesis in infiltrating breast carcinomas.

UNLABELLED: The product of the ATM gene, mutated in the human genetic disorder ataxia-telangiectasia (A-T) plays a key role in the detection and repair of DNA double-strand breaks. A-T is defined by progressive cerebellar ataxia, telangiectasia, sensitivity to ionising radiation and genomic instability with cancer predisposition. On the other hand, increased angiogenesis is essential for tumor growth and metastasis. The aim of this study was to investigate ATM expression in breast carcinomas and its relationship to neoangiogenesis. METHODS AND RESULTS: Fifty-two breast tumors from 51 patients, 38 of them with concomitant in situ component (CIS), were analyzed by immunohistochemistry for the expression of ATM. CD34 expression was used for the morphometric evaluation of vasculature. ATM was positive in 1 to 10% of normal epithelial cells. ATM expression was reduced in 55.8% of infiltrating carcinomas, non-reduced in 34.6%, and increased in 9.6%. Expression of ATM in CIS was similar to the infiltrating component in 71% of cases and reduced in 23.7% of them. High-grade ductal infiltrating carcinomas showed lower ATM expression than low-grade ones. Reduced ATM expression also correlated with increased microvascular area. CONCLUSIONS: Reduced ATM expression in breast carcinomas correlated with tumor differentiation and increased microvascular parameters, supporting its role in neoangiogenesis and tumor progression in breast carcinogenesis.

Clinical axillary recurrence after sentinel node biopsy in breast cancer: a follow-up study of 220 patients.

PURPOSE: The aim of this study was to evaluate the frequency of false-negative (FN) sentinel node procedures in patients with breast cancer and the subsequent clinical outcome in such patients. METHODS: A total of 325 breast cancer patients underwent sentinel lymph node biopsy at our institution between June 1998 and May 2004. A 2-day protocol was used to localise the sentinel node with the injection of 99mTc-nanocolloid. There were two phases in the study: the learning phase (105 patients) and the application phase (220 patients). In the learning phase, a complete lymphadenectomy was always performed. In the application phase, sentinel nodes were studied intraoperatively and lymphadenectomy was performed when considered warranted by the pathological intraoperative results. RESULTS: The median follow-up duration in the 220 patients studied during the application phase was 21.2 months (range 4-45 months). In this phase a total of 427 sentinel nodes were obtained (range 1-5 per patient, median 1.99), with 66 positive sentinel nodes in 56 patients (26%). The lymphadenectomies performed were also positive in 25% of cases (14 patients). We observed a total of two false-negative sentinel lymph node results (3.45%). One of them was found during the surgical excision of non-sentinel nodes, and the other presented as an axillary recurrence 17 months postoperatively (1.72% clinical false-negative rate). The latter patient died 1 year after the first recurrence. CONCLUSION: After a median follow-up of 21.2 months we observed only one clinical recurrence among 220 patients. Our results indicate that adequate local control is achieved by application of the sentinel node protocol.

Interferon-induced depression: prevalence and management.

BACKGROUND: Interferon-induced depression ranges from 0 to 50%. Interferon schedule and a history of psychiatric illnesses are not enough to predict who will develop symptoms and who will not. AIMS: To assess the prevalence of depression during interferon therapy; to test whether Minnesota Multiphasic Personality Inventory is useful in clinical practice for the early identification of patients at risk of depression; whether and how the depression can be cured. PATIENTS: One hundred and eighty-five patients treated with interferon and ribavirin for chronic hepatitis C. METHODS: Before therapy, all patients underwent a Minnesota Multiphasic Personality Inventory and a clinical examination, specifically for the identification of depressive symptoms. RESULTS: Thirty-one patients developed a psychiatric disorder, 11 of them requiring treatment with anti-depressant drugs. Among the 18 patients with Minnesota Multiphasic Personality Inventory positive tests, 16 developed a psychiatric disorder, 8 of them a severe disorder (sensitivity of 0.58; 0.73 for severe disorders). Among the 154 who did not develop psychiatric side effects, 152 had a negative Minnesota Multiphasic Personality Inventory (specificity: 0.99). Severe psychiatric disorders were successfully treated with anti-depressant drugs. CONCLUSIONS: Psychiatric side effects are easy to see during interferon therapy. A psychiatric evaluation should be considered on all patients before treatment. If depression develops, it should be treated aggressively, and selective serotonin re-uptake inhibitors are the anti-depressants of choice.

[Comparative study of the different radiotracer administration route to locate the sentinel node in breast cancer]. / Estudio comparativo de las diferentes vías de administración del radiotrazador para la localización del ganglio centinela en el cáncer de mama.

AIM: To assess the results of Sentinel Lymph Node (SLN) detection in breast cancer patients depending on the radiotracer injection route, especially evaluating which route that makes it possible to identify the highest percentage of SLNs and its detection in the inner mammary chain. METHODS: We prospectively studied 120 breast cancer patients. Three groups of 40 patients each were established depending on the radiotracer injection route. 1) Subdermal or subareolar route: 18 T1 and 22 T2, mean tumor diameter size of 1.76 cm. 2) Peritumoral route: 16 T1 and 24 T2, mean tumor diameter size of 1.86 cm. 3) Intratumoral route: 20 T1 and 20 T2, mean tumor diameter size of 1.61 cm. The day before surgery, a lymphoscintigraphic study was performed in all cases and intraoperative SLN resection was carried out with the aid of a hand-held gamma probe. RESULTS: The SLN was identified in 38/40 cases (95%) of group 1, in 35/40 cases (88%) of group 2 and in 38/40 cases (95%) of group 3. No SLNs in the mammary chain were observed in patients who underwent subdermal/subareolar injection of the radiotracer. On the other hand, SLNs were visualized in this location in 4/35 patients with peritumoral injection (11%) and in 8/38 patients with intratumoral injection (21%). CONCLUSION: The superficial (subdermal, subareolar) injection technique is more suitable when an unnecessary lymphadenectomy has to be prevented in cases without axillary metastases. The deeper injection technique (peritumoral, intratumoral) should be use when, moreover, we seek accurate staging or plan non-systematic mammary chain irradiation.

Estudio comparativo de las diferentes vías de administración del radiotrazador para la localización del ganglio centinela en el cáncer de mama / Comparative study of the different radiotracer administration route to locate the sentinel node in breast cancer

Objetivo: Valorar los resultados de la detección del Ganglio Centinela (GC) en el cáncer de mama dependiendo de la vía de administración del radiotrazador, analizando especialmente qué vía permite identificar el mayor porcentaje de GCs y la detección de GCs en la cadena mamaria interna. Material y métodos: Se estudiaron prospectivamente 120 pacientes con cáncer de mama. Se establecieron tres grupos con 40 casos en cada uno dependiendo de la vía de administración del radiotrazador. 1) Vía subdérmica o subareolar: 18 T1 y 22 T2, diámetro medio de los tumores de 1,76 cm. 2) Vía peritumoral: 16 T1 y 24 T2, diámetro medio de los tumores de 1,86 cm. 3) Vía intratumoral: 20 T1 y 20 T2, diámetro medio de los tumores de 1,61 cm. Se realizó la linfogammagrafía el día previo a la intervención quirúrgica y la resección del GC se llevó a cabo con la ayuda de una sonda de detección externa. Resultados: Se identificó el GC en 38/40 casos (95 %) del grupo 1, en 35/40 casos (88 %) del grupo 2 y en 38/40 casos (95 %) del grupo 3. No se identificaron GCs en la cadena mamaria interna en ninguno de los casos en que se realizó la inyección vía subdérmica/subareolar, mientras que fueron visualizados en 4/35 pacientes con inyección peritumoral (11%) y en 8/38 pacientes con inyección intratumoral (21 %). Conclusión: La técnica de inyección superficial (subdérmica, subareolar) es más adecuada cuando se quiere evitar una linfadenectomía innecesaria en los casos que no presenten metástasis axilares. La técnica de inyección más profunda (peritumoral, intratumoral) debe utilizarse cuando, además, quiera realizarse una estadificación precisa o se plantee la irradiación de la cadena mamaria interna de forma no sistemática (AU)

Aim: To assess the results of Sentinel Lymph Node (SLN) detection in breast cancer patients depending on the radiotracer injection route, especially evaluating which route that makes it possible to identify the highest percentage of SLNs and its detection in the inner mammary chain. Methods: We prospectively studied 120 breast cancer patients. Three groups of 40 patients each were established depending on the radiotracer injection route. 1) Subdermal or subareolar route: 18 T1 and 22 T2, mean tumor diameter size of 1.76 cm. 2) Peritumoral route: 16 T1 and 24 T2, mean tumor diameter size of 1.86 cm. 3) Intratumoral route: 20 T1 and 20 T2, mean tumor diameter size of 1.61 cm. The day before surgery, a lymphoscintigraphic study was performed in all cases and intraoperative SLN resection was carried out with the aid of a hand-held gamma probe. Results: The SLN was identified in 38/40 cases (95 %) of group 1, in 35/40 cases (88 %) of group 2 and in 38/40 cases (95 %) of group 3. No SLNs in the mammary chain were observed in patients who underwent subdermal/subareolar injection of the radiotracer. On the other hand, SLNs were visualized in this location in 4/35 patients with peritumoral injection (11 %) and in 8/38 patients with intratumoral injection (21 %). Conclusion: The superficial (subdermal, subareolar) injection technique is more suitable when an unnecessary lymphadenectomy has to be prevented in cases without axillary metastases. The deeper injection technique (peritumoral, intratumoral) should be use when, moreover, we seek accurate staging or plan non-systematic mammary chain irradiation (AU)

Comparison between local and general anaesthesia for carotid endarterectomy. A retrospective analysis.

AIM: The aim of this study was to compare by a retrospective analysis local anesthesia (LA) versus general anesthesia (GA) for carotid endarterectomy (CEA). METHODS: Two-hundred and 59 patients who underwent CEA, at the Ospedali Riuniti of Reggio Calabria in the period 2000-2001, were enrolled in this study. For analysis purposes, patients were divided into 4 groups, according to their neurological status and to the type of anesthesia. LA was induced either by deep and superficial cervical plexus block (side approach according to Moore). GA was induced with propofol and maintained with sevoflurane. Monitoring of the neurological status was achieved by simple clinical evaluation of the state of consciousness under LA, and by continuous EEG under GA. RESULTS: With LA, a reduction in the duration of intervention and hospital stay, in the number of neurological complications and consequent intraoperative shunts was observed, and admission to the intensive care unit (ICU) was never required. With GA better blood pressure control was achieved, but more intraoperative shunts and admissions to ICU were required, thus increasing overall costs. CONCLUSION: In our experience, LA seems to be the approach of choice for CEA because 1) by preserving the state of consciousness, it allows a simple clinical monitoring of cerebral perfusion maintenance; 2) it reduces the number of intraoperative shunts and the risk of admission to the ICU; 3) it is cost-effective.