Early melanoma invasivity correlates with gut fungal and bacterial profiles.

BACKGROUND: The microbiome is emerging as a crucial player of the immune checkpoint in cancer. Melanoma is a highly immunogenic tumour, and the composition of the gut microbiome has been correlated to prognosis and evolution of advanced melanoma and proposed as a biomarker for immune checkpoint therapy. OBJECTIVES: We investigated the gut fungal and bacterial compositions in early-stage melanoma and correlated microbial profiles with histopathological features. METHODS: Sequencing of bacterial 16S rRNA and the fungal internal transcribed spacer region was performed on faecal samples of patients with stage I and II melanoma, and healthy controls. A meta-analysis with gut microbiota data from patients with metastatic melanoma was also carried out. RESULTS: We found a combination of gut fungal and bacterial profiles significantly discriminating patients with melanoma from controls. In patients with melanoma, we observed an abundance of Prevotella copri and yeasts belonging to the order Saccharomycetales. We found that the bacterial and fungal community correlated to melanoma invasiveness, whereas the specific fungal profile correlated to melanoma regression. Bacteroides was identified as general marker of immunogenicity, being shared by regressive and invasive melanoma. In addition, the bacterial communities in patients with stage I and II melanoma were different in structure and richer than those from patients with metastatic melanoma. CONCLUSIONS: The composition of the gut microbiota in early-stage melanoma changes along the gradient from in situ to invasive (and metastatic) melanoma. Changes in the microbiota and mycobiota are correlated to the histological features of early-stage melanoma, and to the clinical course and response to immune therapies of advanced-stage melanoma, through direct or indirect immunomodulation.

ECG-gated CT angiography of the thoracic aorta: the importance of evaluating the coronary arteries.

AIM: To evaluate the feasibility of coronary artery disease (CAD) evaluation using electrocardiogram-gated computed tomography CT of the thoracic aorta. MATERIALS AND METHODS: A total of 477 patients, who underwent CT angiography of the thoracic aorta, were included retrospectively. Dose-length products (DLP) were recorded. Two blinded readers graded image quality of the coronary arteries on a three-point scale. Coronary artery stenosis has only been reported if considered significant, i.e., ≥50%. The type of plaque responsible for the stenosis was considered. The normal distribution of the data was assessed using Shapiro-Wilk and Anderson-Darling tests. Results were expressed as means and standard deviations and percentages. Inter-reader agreements were analysed by calculating the intraclass correlation coefficient, and by using Cohen kappa statistics. RESULTS: The mean DLP was 566±90.4 mGy∙cm, corresponding to an effective dose of 9.6±1.5 mSv. Five point three percent of asymptomatic patients were positive for CAD with stenosis ≥50%. All patients with coronary stenosis presented with a soft plaque. Two anomalous coronary origins were found. The inter-reader agreement was excellent in defining both the quality of the examination and the degree of coronary stenosis (k=0.85). CONCLUSION: The opportunity to prove the presence of CAD in asymptomatic patients during a ECG-gated CT of the thoracic aorta can have an extremely important clinical impact, promoting the best therapeutic pathway for the patient. Therefore, coronary arteries should always be analysed carefully and reported in ECG-gated CT angiography of the thoracic aorta.

Staged hybrid treatment of complex ascending aortic and distal aortic arch pseudoaneurysm after repair of aortic coarctation.

A 49-year-old operated for aortic coartaction patient presented with thoracic and ascending aortic aneurysm. He was asymptomatic. Angio-magnetic resonance nuclear scan and angiography revealed an ascending aortic aneurysm (5.2 cm), bicuspid aortic valve, 6-cm proximal descending aortic pseudoaneurysm at the site of the previous operation with involvement of the left subclavian artery. Restenosis at the original site of coarctation and aortic arch hypoplasia distally to the brachiocefalic trunk was also found. The operation performed was a "modified Bentall - De Bono". The pseudoaneurysm was not accessible through median sternotomy due to the massive lung adhesions following the previous surgery. The left common carotid artery was explanted from the aortic arch and connected with a graft to the ascending aortic conduit. A proximal neck suitable for landing zone of the endovascular stent-graft was then established. The postoperative course was uneventful. After two weeks, the patient was readmitted. The exclusion of the thoracic descending aortic pseudoaneurysm by endovascular implantation of the stent-graft prosthesis was performed. The left subclavian artery was excluded because left vertebral artery was closed. The patient did not develop hand claudicatio. The procedure was successful.

Different carbon sources affect lifespan and protein redox state during Saccharomyces cerevisiae chronological ageing.

In this study, a proteomic approach that combines selective labelling of proteins containing reduced cysteine residues with two-dimensional electrophoresis/mass spectrometry was used to evaluate the redox state of protein cysteines during chronological ageing in Saccharomyces cerevisiae. The procedure was developed on the grounds that biotin-conjugated iodoacetamide (BIAM) specifically reacts with reduced cysteine residues. BIAM-labelled proteins can then be selectively isolated by streptavidin affinity capture. We compared cells grown on 2% glucose in the exponential phase and during chronological ageing and we found that many proteins undergo cysteine oxidation. The target proteins include enzymes involved in glucose metabolism. Both caloric restriction and growth on glycerol resulted in a decrease in the oxidative modification. Furthermore, in these conditions a reduced production of ROS and a more negative glutathione half cell redox potential were observed.

Genomics approach to the analysis of bacterial communities dynamics in Hirschsprung's disease-associated enterocolitis: a pilot study.

INTRODUCTION: The most invalidating and life-threatening complication in Hirschsprung's disease patients (HSCR) is Hirschsprung's disease-associated enterocolitis (HAEC). The mechanisms underlying enterocolitis have not been identified. The limited knowledge of the role of intestinal microflora is in part due to the complexity of the intestinal microbiome and to the limitation of cultivation-based technologies, given that less than 25% of the intestinal bacterial species can be cultured. MATERIALS AND METHODS: We used amplified ribosomal DNA restriction analysis (ARDRA) with four different restriction enzymes to study variations of microflora composition of the stools of a selected HSCR patient in different clinical conditions (acute phase vs. remission). RESULTS: We assessed a total of 15 stool specimens belonging to the same 3-year-old male patient suffering from HSCR, which were harvested during 4 HAEC episodes and remission phases. Restriction analysis showed that HAEC episodes seem to cluster together at ARDRA analysis, thus suggesting a sort of predisposing bacterial community for HAEC development and the need for a microflora equilibrium to maintain wellness. CONCLUSIONS: This approach proved to be effective, useful and powerful in assessing microflora dynamics and indicated that the differences in microflora associated with acute HAEC or remission are likely to result from a combination of disease activity and different antibiotic therapies. ARDRA proved to be useful in discriminating disease versus remission. Our findings indicated that HAEC results from a change in the equilibrium between bacterial species or from altered discrimination of harmless from harmful microorganisms, challenging the definition of pathogenic and non-pathogenic species. Based on these results, we propose ARDRA as a rapid inexpensive tool to assess microflora dynamics during HAEC episodes.

Nod2 Deficiency in mice is Associated with Microbiota Variation Favouring the Expansion of mucosal CD4+ LAP+ Regulatory Cells.

Nucleotide-binding Oligomerization Domain-2 (NOD2) mutations are associated with an increased risk to develop Crohn's Disease. In previous studies, we have shown that Nod2-/- mice manifest increased proportion of Lamina Propria (LP) CD4+ LAP+ Foxp3- regulatory cells, when compared with Nod2+/+ mice, while CD4+ Foxp3 + regulatory cells were not affected. Here, we investigated the Nod2 gut microbiota, by 16S rRNA pyrosequencing, at steady state and after TNBS-colitis induction in mice reared separately or in cohousing, correlating the microbial profiles with LP regulatory T cells proportion and tissue cytokines content. We found that enrichment of Rikenella and Alistipes (Rikenellaceae) in Nod2-/- mice at 8 weeks of age reared separately was associated with increased proportion of CD4+ LAP+ Foxp3- cells and less severe TNBS-colitis. In co-housed mice the acquisition of Rickenellaceae by Nod2+/+ mice was associated with increased CD4+ LAP+ Foxp3- proportion and less severe colitis. Severe colitis was associated with enrichment of gram-negative pathobionts (Escherichia and Enterococcus), while less severe colitis with protective bacteria (Barnesiella, Odoribacter and Clostridium IV). Environmental factors acting on genetic background with different outcomes according to their impact on microbiota, predispose in different ways to inflammation. These results open a new scenario for therapeutic attempt to re-establish eubiosis in Inflammatory Bowel Disease patients with NOD2 polymorphisms.

Microsatellite instability in a population of sporadic colorectal cancers: correlation between genetic and pathological profiles.

BACKGROUND: Tumours with high-frequency microsatellite instability exhibit unique genotype and phenotype features, whereas the difference between low-frequency microsatellite instability and apparently stable tumours is far from being clear. AIMS: To identify distinctive genetic and pathological characteristics of low-frequency microsatellite instability tumours. METHODS: Microsatellite instability status of 57 sporadic colorectal cancers and its correlation with genetic, pathological and clinical features was analysed. RESULTS: High frequency microsatellite instability and low-frequency microsatellite instability and apparently stable cancers were different in terms of tumour localisation (p=0.015), frequency of APC mutations (p=0.012), occurrence of Crohn's-like/lymphoid reaction (p=0.0353) and morphological evidence of origin from an adenoma (p=0.0338). Specifically, in low-frequency microsatellite instability cancers, APC mutations were very frequent (76.9%, 10/13) and a Crohn's-like/lymphoid reaction was common (38.5%, 5/13). High-frequency microsatellite instability tumours were preferentially located in the right colon and exhibited a higher frequency of loss of heterozygosity at the FHIT locus compared with low-frequency microsatellite instability and apparently stable cases (p=0.0243). Dukes' stage (p=0.0021), tumour localisation (p=0.0410) and pattern of cancer growth (p=0.0374), were the only factors affecting patient survival. However, a borderline improvement was noted in overall survival in high-frequency microsatellite instability and low-frequency microsatellite instability cancer patients (p=0.062). CONCLUSIONS: These results indicate that low-frequency microsatellite instability tumours have different genetics and histological features and suggest that they are a distinct group of colorectal cancers.

The use of mechanical prostheses in native aortic valve endocarditis.

BACKGROUND AND AIMS OF THE STUDY: The optimal aortic valve substitute in cases of active native valve endocarditis (NVE) remains controversial. This report summarizes our experience with the surgical treatment of active aortic NVE using only mechanical prostheses. METHODS: Between January 1988 and January 1996, 20 patients underwent aortic valve replacement for active NVE. There were 17 men and three women. Mean age was 46.5 years (range eight to 63 years). Thirteen patients were in NYHA class IV and seven in class V. Streptococci were isolated in eight cases, while no causative micro-organism could be identified in seven patients. All operations were performed on urgent (n = 13) or emergency (n = 7) bases. A mechanical valve was implanted in all cases and radical resection of the infected tissues performed using different techniques. All patients were followed up at our institution. Two-dimensional color Doppler studies were performed one month after surgery and at six-month intervals after the first year. Transesophageal echocardiography (TEE) was performed at discharge, six months after surgery and yearly thereafter. RESULTS: No patient died in hospital. Mean follow up was 30.5 months, during which time three patients died, though none from endocarditis-related causes. Endocarditis recurred only one (5%). TEE demonstrated a normally functioning aortic prosthesis in 15 cases and trivial paravalvular leakage in two. CONCLUSIONS: Mechanical prostheses represent a safe aortic valve substitute in cases of acute native valve endocarditis. When radical resection of all the infected areas is performed, the incidence of endocarditis recurrence is acceptable. The concept that homografts are the valve substitute of choice in endocarditis cases cannot be supported by this study.

Inhibition of 1,2-dimethylhydrazine-induced oxidative DNA damage in rat colon mucosa by black tea complex polyphenols.

The effect of black tea polyphenols on 1,2-dimethylhydrazine (DMH)-induced oxidative DNA damage in rat colon mucosa has been investigated. Fischer 344 rats were treated orally with thearubigin (TR) or theafulvin (TFu) for 10 days (40 mg/kg), injected ip with DMH (20 mg/kg) or saline and sacrificed 24 hr after DMH administration. The levels of 8-hydroxy-2'-deoxyguanosine (8-OHdG) were measured in colonic mucosa DNA and expressed as a ratio relative to 2'-deoxyguanosine (2dG). Control rat mucosa had 8-OHdG values of 1.12 +/- 0.14/10(5) dG (mean +/- SEM, n=11), whereas DMH-treated rats significantly higher values (1.52 +/- 0.14/10(5) dG, n=26, P<0.05). Pretreatment of rats with TR had significantly inhibited DMH-induced oxidative DNA damage 0.99 +/- 0.09/10(5) dG, n=10, P<0.05) and a similar, although less marked, effect was observed with TFu (1.15 +/- 0.19/10(5), n=9, P=0.06). These findings confirm that DMH causes oxidative DNA damage in the colon mucosa of rats and demonstrate that this effect is prevented by the consumption of complex polyphenols from black tea.

Photopheresis in cutaneous T-cell lymphoma: five-year experience.

BACKGROUND: Cutaneous T-cell lymphoma (CTCL) includes several lymphoproliferative disorders involving mature T-lymphocyte proliferation initially confined to the cutis. These affections, after variable periods, may progress to the blood, limph nodes and visceral organs. Mycosis fungoides (MF) is the most frequent form of CTCL and has an indolent clinical course. The therapy of CTCL depends on the stage of the disease and the patient's general conditions. For advanced cases it includes chemotherapy, retinoids, and interferon-alpha. Since 1987 extracorporeal photochemotherapy (ECP), a novel immunomodulatory approach based on apheresis and photoirradiation of leukocytes, has been successfully introduced for the treatment of advanced CTCL. It can prolong survival of patients with erythrodermic CTCL without significant side effects. OBJECTIVE: To review our five-year experience with ECP in CTCL. METHODS: Since June 1994, 33 CTCL patients have been recruited for ECP, using two different regimens: two procedures on two consecutive days at four-week intervals for six months, or at two-week intervals for three months with progressive tapering in the second three-month period for the more severe forms. Six patients received ECP with IFN-alpha. ECP was done using the photopheresis UVAR system and UVAR XTS (Therakos, West Chester, Pa) and always with 8-MOP liquid formulation injected directly into the buffy coat bag. Lymphocytes in peripheral blood were immunophenotypically characterized for each patient and every ECP session. RESULTS: All patients tolerated ECP well, without significant side effects. Thirty patients are clinically evaluable (at least three ECP cycles). A favourable clinical response was obtained in 80.9% (16/21) of MF patients (complete response 33%, partial response 47.6%) and in 66% (6/9) of patients in the Sézary's syndrome phase (complete response 33.3%, partial response 33.3%). Five of the six patients given IFN-alpha as adjunctive therapy had a PR and one a CR. Four patients are in CR without therapy at follow-ups of 46, 20, 10 and 8 months. There have been no changes in the peripheral lymphocyte immunophenotype during the follow-up. In 19/30 patients the CD95 antigen, correlated with cellular apoptosis, was expressed and was frequently associated with a good clinical response. CONCLUSIONS: In our experience ECP achieved favourable clinical responses in 73% of patients, in monotherapy or in combination with IFN-alpha, without significant side effects.

Deriving passage difficulties for a tracking study via the Cloze technique.

The current report demonstrates the importance of formally accounting for passage difficulty when using the tracking procedure. Cloze responses to 82 encyclopedia excerpts (343-349 words each) were obtained from a large pool of normal-hearing adults and scored verbatim. Passage difficulty, derived via ANOVA, was then defined as the deviation of a passage's mean Cloze score from the score for all passages, corrected for differences among respondents. The passage difficulties were applied in an alternating conditions tracking experiment with one adult cochlear implant user. Conditions included conventional auditory-visual and auditory-only tracking and experimental mode-switching techniques in which the talker changed modalities during the correction phase. An ANCOVA of the word-per-minute scores was conducted, with passage difficulty as a covariate and passage adjustment values as the output. Tracking rates and percentage of words correct from the beginning and end of training were examined. Use of adjusted data reversed the interpretation of performance change, demonstrating the need for determining passage difficulties a priori.

[Increase of beta 2-microglobulin in drug addicts with anti-HIV antibodies and high risk of AIDS]. / Incremento della beta 2-microglobulina in soggetti tossicodipendenti con anticorpi anti-HIV e ad alto rischio per AIDS.

The beta 2-Microglobulin is a polypeptide present on the surface membrane of both B and T cells and is integrated into the structure of HLA antigenes. The beta 2-Microglobulin concentration have been used as a reliable indicator of glomerular and tubular function of the kidney. Increased serum concentration of beta 2-Microglobulin are observed also in lymphoproliferative disorders with high cell proliferation rates. More recently, increased concentration of beta 2-Microglobulin was shown in patients with anti-HIV antibodies with or without symptomatic AIDS. We have determined beta 2-Microglobulin in 61 subjects: 40 between the ages of 25 and 35 and seemingly healthy, 21 patients between the ages of 22 and 32 and intravenous drug abuser with anti-HIV antibodies and at high-risk for AIDS. In all subjects we have tested: BUN, creatinine, beta 2-Microglobulin and T4/T8 ratio. In 40 subjects as normal controls, beta 2-Microglobulin average was means = 1.07 mg/L (SD = 0.39), T4/T8 ratio average: means = 1.06 (SD = 0.119). In 21 patients drug abuser with anti-HIV antibodies, the beta 2-Microglobulin average was cleanly increased: means = 4.72 mg/L (SD = 2.23), the T4/T8 ratio average cleanly decreased: means = 0.54 (SD = 0.21). We believe the beta 2-Microglobulin quantitation, even if not specific for patient with symptomatic AIDS, used in conjunction with other laboratory tests, principally T4/T8 ratio, will be a useful marker for recognizing persons with possible asymptomatic AIDS who are members of populations known to be at high-risk for AIDS.

[Reintervention of myocardial revascularization by left thoracotomy in deep hypothermia. A clinical case]. / Reintervento di rivascolarizzazione miocardica per toracotomia sinistra in ipotermia profonda. Caso clinico.

Herewith we report a case of redo coronary-surgery which was approached through a left thoracotomy and in hypothermic arrest so as to avoid either a demanding dissection with its potential consequences over the functioning grafts and aortic cross-clamping thus making unnecessary any cardioplegia delivery. We describe the surgical procedure and analyze its advantages with respect to the usual median sternotomy approach.

Use of ambiguous visual stimuli to demonstrate the value of acoustic cues in speech perception.

The purpose of this study was to evaluate a set of English phrases and sentences in a minimal-pairs syllable-test format for assessing use of acoustic cues in the audiovisual perception of speech; in particular, by persons with severe or profound hearing loss. Test items that were not rejected were categorized as "acceptable" or "ideal." An acceptable item was visually confusable but auditorally distinct compared to its foil (analogous to /p/vs/m/). An ideal item was, in addition, tactilely distinct. Forty-eight items, spoken by a male and a female talker, were presented in a videotaped two-alternative force-choice format under three conditions to three young-adult subject groups. In a lipreading-without-sound condition, 39 items were visually confusable; of these, 16 items were reliably identifiable when sound was added (the "acceptable" items). Seven of the 16 acceptable items were also identifiable in a tactile-pattern-alone condition (the "ideal" items). Performance on the 16 acceptable items was analyzed for an additional 27 young adults with moderate, severe, or profound hearing loss. Results obtained with the new protocol were consistent with conventional assessment results (CID Everyday Sentences, with and without sound) in all but 4 cases. Communication characteristics of the exceptions were examined to explain lack of agreement.

[Lesions of the inferior alveolar nerve]. / Lesioni nervose a carico del nervo alveolare inferiore.

All possible forms of inferior alveolar nerve injury are examined with a report on several personal cases and their medicolegal implications.

[Histiocytic necrotizing lymphadenitis or Kikuchi and Fujimoto's disease. A case report]. / Linfoadenite necrotizzante istiocitaria o malattia di Kikuchi e Fujimoto. Descrizione di un caso.

We describe a case of necrotizing histiocytic lymphadenitis or Kikuchi and Fujimoto disease, a rare lymphadenopathy of unknown etiology usually striking young women. It was formerly described in Japan, and subsequently in other countries and in Italy as an anatomo-clinical entity. The disease has a favourable prognosis, therefore it has to be considered differently from other lymphadenopathies with severe prognosis whose anatomo-pathological differential features were analyzed by the authors. The case here observed differs from other cases described in literature because of the presence of a perilymphadenitic component.

Cirugía de control de daños: ¿que hay de nuevo? / Damage control surgery: what's new?

Antecedentes. La cirugía de control de daños se ha ganado un espacio en el tratamiento de un pequeño número de enfermos con traumatismos graves. Objetivo. Actualizar los conocimientos que se han aportado para perfeccionar esta táctica clínico-quirúrgica. Lugar de aplicación. C.M.P.F. "Churruca-Visca" Diseño. Análisis de los nuevos conceptos desarrollados en el último decenio. Conclusiones. Este tema ha sido actualizado con la intención de exponer los nuevos avances en la resucitación, el traslado y el sostén del medio interno de estos enfermos. Esta táctica médico-quirúrgica se ha puesto en boga para un selecto grupo de pacientes que reúnen condiciones excepcionales y probables chances de muerte. El control del daño en fase cero obliga a una resucitación más dinámica en el lugar del hecho y durante el traslado. El control del daño resuscitatorio involucra un nuevo concepto basado en un antiguo precepto de la cirugía del aneurisma complicado "la resucitación hipotensiva". El manejo de las lesiones arteriales y venosas son la llave del éxito de este procedimiento. El uso de la arteriografía selectiva las 24 hs permiten rescatar muchos enfermos con lesiones de difícil abordaje.(AU)

Laboratory projects in tactile aids to lipreading.

Four studies are reported on the design of tactile aids to lipreading conducted at Central Institute for the Deaf, St. Louis. A speech feature aid represented resonance, envelope, and source characteristics of speech on three vibrators at the fingertips. Artificially-deafened and congenitally deaf adults lipreading with the aid obtained an average of 40% gain over lipreading alone on syllable identification tests, but minimal gain for untrained words. In a series of three other studies, the quality of tactile sensation was developed as a cue to spectral information, presented in a temporal code based on the overall time-intensity waveform of speech. Electrical and vibratory stimulation on the hand and torso were evaluated by artificially-deafened adults using the tracking technique with connected speech. Short-term training yielded variable increments over unaided lipreading scores and lipreading with a simple vibrator aid. Neither the feature nor the spectral code was superior in aiding lipreading. The displays that produced quality changes seemed more efficient for training than the spatial display. Results were affected also by content and method of training and initial lipreading proficiency.

A method for training and evaluating the reception of ongoing speech.

The "tracking procedure" is a method to train and evaluate the reception of ongoing speech. A talker and a receiver engage in a dialogue for a designated period of time in which the receiver reports his perception of successive segments of read text and is corrected by the talker until the text is repeated verbatim. Performance is measured in number of words of text repeated correctly per unit of time. The procedure is illustrated by its application to a project for training and evaluating use of a vibrotacile-electrotactile aid to lipreading. Differences between experimental conditions are described over time in terms of absolute words per minute, increment of aided over unaided words-per-minute scores, and percent of normal listening rate. Data from identification tests with syllables, words, and sentences are also presented. Tracking is suggested as an additional procedure for communication training in aural rehabilitation or classroom setting, and for evaluation of other communication systems.