A novel BRAF mutation in association with primary amelanotic melanoma with oral metastases.

BACKGROUND: In the context of amelanotic melanoma, little is known on the genetic or molecular background that determines the onset of this peculiar phenotype of melanoma and its sites of metastatic spread. However, it appears that amelanotic melanomas frequently lack BRAF mutations. OBJECTIVE: To report the genetical analysis of one case amelanotic melanoma developing oral metastasis. METHODS: The BRAF mutational status of the primary lesion was assessed by both Sanger sequencing and pyrosequencing. RESULTS: Both methodologies showed changes in three nucleotides: C1796T; G1798A and T1799A. These mutations should result in a rare double aminoacid substitution in codons 599 and 600 of the BRAF protein (BRAF T599I/V600K). CONCLUSION: This unusual mutation was associated with an uncommon clinical phenotype of the primary tumour and with an unusual site of metastatic spread. In the lack of comparable data, a potential association between the unusual mutation and clinical findings remains a matter of further studies.

Discovery of a transient absorption edge in the X-ray spectrum of GRB 990705.

We report the discovery of a transient equivalent hydrogen column density with an absorption edge at approximately 3.8 kiloelectron volts in the spectrum of the prompt x-ray emission of gamma-ray burst (GRB) 990705. This feature can be satisfactorily modeled with a photoelectric absorption by a medium located at a redshift of approximately 0.86 and with an iron abundance of approximately 75 times the solar one. The transient behavior is attributed to the strong ionization produced in the circumburst medium by the GRB photons. The high iron abundance points to the existence of a burst environment enriched by a supernova along the line of sight. The supernova explosion is estimated to have occurred about 10 years before the burst. Our results agree with models in which GRBs originate from the collapse of very massive stars and are preceded by a supernova event.

Cadherin-6 promotes EMT and cancer metastasis by restraining autophagy.

The transdifferentiation of epithelial cells toward a mesenchymal condition (EMT) is a complex process that allows tumor cells to migrate to ectopic sites. Cadherins are not just structural proteins, but they act as sensors of the surrounding microenvironment and as signaling centers for cellular pathways. However, the molecular mechanisms underlying these signaling functions remain poorly characterized. Cadherin-6 (CDH6) is a type 2 cadherin, which drives EMT during embryonic development and it is aberrantly re-activated in cancer. We recently showed that CDH6 is a TGFß target and an EMT marker in thyroid cancer, suggesting a role for this protein in the progression of this type of tumor. Papillary thyroid carcinomas (PTCs) are usually indolent lesions. However, metastatic spreading occurs in about 5% of the cases. The identification of molecular markers that could early predict the metastatic potential of these lesions would be strategic to design more tailored approaches and reduce patients overtreatment. In this work, we assessed the role of CDH6 in the metastatic progression of thyroid cancer. We showed that loss of CDH6 expression profoundly changes cellular architecture, alters the inter-cellular interaction modalities and attenuates EMT features in thyroid cancer cells. Using a yeast two-hybrid screening approach, based on a thyroid cancer patients library, we showed that CDH6 directly interacts with GABARAP, BNIP3 and BNIP3L, and that through these interactions CDH6 restrains autophagy and promotes re-organization of mitochondrial network through a DRP1-mediated mechanism. Analysis of the LIR domains suggests that the interaction with the autophagic machinery may be a common feature of many cadherin family members. Finally, the analysis of CDH6 expression in a unique cohort of human PTCs showed that CDH6 expression marks specifically EMT cells. and it is strongly associated with metastatic behavior and worse outcome of PTCs.

Contemporary and potential future molecular diagnosis of melanoma.

INTRODUCTION: The increasing incidence of cutaneous melanoma and the still limited effective treatments available for this disease represent a major health problem and a great challenge for research. The raise of the "omics" era and the development of new techniques to explore phenotypic heterogeneity are helping to decipher the mechanisms at the basis of melanoma heterogeneity. AREAS COVERED: We reviewed the most recent publications about the biology of cutaneous melanoma, to provide an overview of the most recent insights into the complexity of this tumor and their potential impact in the clinical settings. Expert commentary: Starting from the first attempts to provide a molecular classification of melanoma, it has been evident that this tumor represents a widely heterogeneous disease. This complexity and the multivariate nature of melanoma represent a major obstacle in developing the best management strategies for patients.

Chlorinated hydrocarbons in fish from the river Po (Italy).

Pesticides and PCBs levels have been determined in two species of fish from the river Po during 1979--1980 at five sampling stations in different seasons. Significant higher levels have been observed in most industrial areas. When expressed on the basis of fat content, river fish levels are higher than those reported for fish from the Adriatic sea. Using experimental BCF (bioconcentration factors), pesticides and PCB concentrations in water have been calculated from fish values, and compared with measured concentrations.

[Endobronchopulmonary therapy with a permanent trans-crico-thyroid probe]. / Terapia endobroncopolomonare con sonda permanente trans-crico-tiroidea.

The possibilities of treating chronic bronchopneumopathies of various nature by means of permanent endobronchial transcricothyroid probe are examined with reference to 6 cases representing patients under treatment at the time this paper was being prepared (2 cases of chronic pulmonary TB, 2 of suppurating bronchiectasis, 1 of middle lobe syndrome and 1 of tracheal carcinoma). The series examined confirms the usefulness of this type of treatment as a back-up to traditional medical treatment where this alone failed to achieve the necessary results. Stress is laid on the good tolerance of the TCT probe presented by all patients, including a five-year-old boy.

[Cryotherapeutic destruction of invasive tracheo-bronchial tumors. Personal case histories]. / Disostruzione crio-terapica dei tumori invasivi tracheo-bronchiali. Casistica personale.

Data are presented on 15 cases of invasive tracheobronchial tumours subjected to cryotherapy in 1984-85. The technique is indicated in patients who cannot be given surgical or radiation treatment and in cases of asphyxial syndrome requiring faster deobstruction than is obtainable with radiation treatment.

[Abnormal left pulmonary artery: a cause of severe newborn cardiorespiratory insufficiency]. / Arteria polmonare sinistra aberrante: una causa di insufficienza cardiorespiratoria neonatale grave.

Aberrant left pulmonary artery (vascular sling) indicates the vascular anomaly in where the left pulmonary artery arises from the right pulmonary artery, passes over the right mainstem bronchus, loops to the left and courses between the trachea and esophagus to the left hilus. Almost invariably respiratory symptoms result from compression of the trachea and right mainstem bronchus by the anomalous vessel as well as by associated tracheobronchial defects. A case is reported. The distinctive radiologic features include anterior indentation of the barium esophagogram and a characteristic pulmonary angiogram. Surgery is the only resolutive treatment for symptomatic patients, but results are often poor owing to associated intracardiac defects and intrinsic tracheobronchial anomalies.

[Traumatic intramural hematoma of the duodenum. Description of a case]. / Ematoma intramurale traumatico del duodeno. Descrizione di un caso.

The traumatic intramural hematoma of the duodenum is the result of contusion of the retroperitoneal portion of the duodenum. This lesion occurs most frequently in children and can result in complete obstruction of the second and third portion of the duodenum.

[Umbilical cord IgE: a test for sensitivity, specificity and predictive value of atopy?]. / IgE cordonali: test di sensibilità, specificità e predittività di atopia?

Total IgE cord serum levels were measured in 200 newborns in the Hospital of Viadana-Bozzolo and familial allergic conditions were investigated in all the cases. In a 6 month follow up study attention was focused on the development of atopic symptoms. The sensibility, specificity and predictive value of the total IgE cord serum levels are discussed and the introducing of preventive measures in the at risk newborns is also questioned.

Effect of free thymol on differential gene expression in gastric mucosa of the young pig.

Thymol is the most common molecule in thyme and has been proposed as an oral alternative to antibiotics in the feed of pigs and broilers. The knowledge of the in vivo physiological effects of thymol on tissues is limited, particularly its impact on the gastric mucosa, where it is primarily absorbed when it is orally supplied. In this study, thymol (TH, 50 mg/ kg BW) or a placebo (CO) was introduced directly into the stomach of 8 weaned pigs that were slaughtered 12 h later and sampled for gastric oxyntic and pyloric mucosa. The analysis of whole transcript expression was performed using Affymetrix© Porcine Gene 1.1 ST array strips. Affymetrix Transcripts IDs were associated with 13 406 human gene names based on Sus scrofa Ensemble. Gene Set Enrichment Analysis was performed, comparing TH and CO pigs. For each gene set, the normalized enrichment score (NES) was defined as significant when the false discovery rate % was <25 and the P-value of NES was <0.05. In response to TH, 72 and 19 gene sets were significantly enriched in the oxyntic and pyloric mucosa, respectively. Several gene sets involved in mitosis and its regulation ranked near the top, primarily in the oxyntic mucosa; the gene set DIGESTION ranked first and ninth in the pyloric and oxyntic mucosa, respectively. Within this group, somatostatin (SST), SST receptors, peptide transporter 1 (SLC15A1) and calpain 9 (gastrointestinal tract-specific calpain) were the most strongly upregulated genes. Thymol reduced the enrichment of 120 and 59 gene sets in the oxyntic and pyloric mucosa, respectively. Several gene sets related to ion transport and channeling and aqueous pores across membranes, including short transient receptor potential (TRP) channel 4, potassium voltage-gated channel members 1 and 2, and ryanodine receptors 2 and 3, were less enriched. The downregulation of these genes sensitive to thymol in vitro could depend on the thymol dose and contact with the gastric tissues that causes an adaptive response with their reduced activation. Conversely, the activation of the TRPA1 gene (ranked 1072 and 128 among all the genes in the oxyntic and pyloric mucosa, respectively) indicates the involvement of another TRP-regulating cellular calcium storage. In conclusion, the stimulation of gastric proliferative activity and the control of digestive activity by thymol can influence positively gastric maturation and function in the weaned pigs. These properties should be considered in addition to thymol's antimicrobial properties when supplementation of this molecule in feed is evaluated.

Assessment of the presence of chemosensing receptors based on bitter and fat taste in the gastrointestinal tract of young pig.

Knowledge on porcine bitter and fat taste receptors and on their expression in gastrointestinal tract of pigs is scarce. We searched for the presence of porcine homologous sequences for 13 human transcripts of bitter and fat taste receptors in ENSEMBL and National Center for Biotechnology Information databases. For taste 2 receptor (TAS2R) 8, alignment was not observed; for TAS2R13 and TAS2R46 the porcine predicted sequence aligned with several other human bitter genes. For 7 genes for bitter taste (TAS2R1, TAS2R3, TAS2R7, TAS2R9, TAS2R10, TAS2R16, and TAS2R38) and for 3 genes for fat taste (GPR40, GPR43, and GPR120), a full homology for exon sequences was found and primers were designed by Primer3. These 7 genes were amplified with real-time PCR and verified on agarose gel in 5 gastrointestinal segments of weaned pigs: oxyntic (ST1), pyloric (ST2), and cardiac to oxyntic transition mucosa (ST3), jejunum (JEJ), and colon (COL). Suitability of mRNA was verified by amplifying RPL4 and HMBS2 genes. Each bitter taste gene was detectable on agarose gel in at least 1 subject of all the gastrointestinal segments except for TAS2R3 and TAS2R38 that were never detected in ST1 and COL, respectively. The inspection of bitter taste genes amplification curve indicated that the expression was in general very low. GPR43 and GPR120 were present in all segments from all pigs. Expression was not detected for GPR40. Data also indicate that colon is the preeminent tract where fat detection by GPR120 takes place (P < 0.001). The presence of gene expression for several chemosensing receptors for bitter and fat taste in different compartments of the stomach confirms that this organ should be considered a player for the early detection of bolus composition.

In vitro test on the ability of a yeast cell wall based product to inhibit the Escherichia coli F4ac adhesion on the brush border of porcine intestinal villi.

The ability of a yeast cell wall (YCW)-based product (SENTIGUARD C; Nutriad) to inhibit the enterotoxigenic Escherichia coli F4ac (ETEC) adhesion on the brush border of porcine intestinal villi was tested. The ETEC suspensions were preincubated with 2 batches of the product (A and B) at different concentrations (10, 5, and 0.5%, wt/vol) or with their filtrates (AF and BF) and then with intestinal villi susceptible to ETEC adhesion. In all the trials, ETEC suspensions were also preincubated with egg yolk (E) immunized against ETEC to assess the maximum inhibition of the adhesiveness or directly with the villi [control group (Con)] to verify the maximum adhesiveness of the pathogen. For each treatment, 20 different villi were observed, brush border measured, and the adherent pathogens counted. A scanning electron microscope analysis was used to confirm the ability of ETEC to adhere on the YCW. The E treatment significantly reduced the pathogen adhesion on the villi compared with the C group in all the trials (P < 0.001). Both batches of SENTIGUARD C significantly reduced the pathogen adhesion on the villi compared with the C group at the concentration of 10 and 5% (P < 0.001) but not at the concentration of 0.5%. The BF did not significantly reduce the ETEC adhesion whereas the AF significantly increased bacterial adhesion (P = 0.015). The microscopy results confirm the ability of ETEC to adhere on YCW. Taken together, our results indicate the ability of the SENTIGUARD C to contain the intestinal infection from ETEC in young pigs with the affinity of ETEC to YCW.

Investigation on CAST, CAPN1 and CAPN3 porcine gene polymorphisms and expression in relation to post-mortem calpain activity in muscle and meat quality.

This study aimed to detect variability in CAST, CAPN1 and CAPN3 porcine genes and to investigate the effect of CAST and CAPN1 polymorphisms on the activity of native and autolyzed µ-calpain and m-calpain, measured from 1 to 72 h post-mortem in Longissimus dorsi (LD) muscle of 30 pigs. Effects of polymorphisms on meat quality parameter such as pH, color and drip loss were also evaluated. Samples carrying CAST EU137105:g.76,872AA genotype showed higher autolyzed µ-calpain activity 24 and 72 h post-mortem, as well as lower drip loss values. Expression of CAST, CAPN1 and CAPN3 was assessed in LD muscles divergent for shear force. Higher CAST and CAPN3 expression was found in LD with high shear force (P<0.2), confirming a direct role for calpastatin but not for calpain 3 in meat tenderization. In conclusion, CAST gene affected post-mortem activation time of calpain and drip loss.

Perilipin 1 and perilipin 2 protein localization and gene expression study in skeletal muscles of European cross-breed pigs with different intramuscular fat contents.

This study investigated the lipid droplet coat proteins perilipin 1 (PLIN1) and perilipin 2 (PLIN2) localization in pig skeletal muscle and their relationship with intramuscular fat (IMF) content. PLIN1 and PLIN2 proteins were immunostained in semimembranosus muscle cross-sections from two groups of samples divergent for IMF and the gene expression was quantified. PLIN1 localized in the periphery of intramuscular adipocytes, whereas PLIN2 localized within myofibers with high lipid content. The high IMF group showed higher total cross-sectional area of PLIN1-stained adipocytes compared with the low IMF group (P<0.05), while the cross-sectional area and percentage of PLIN2-positive myofibers did not differ between IMF-divergent groups. This suggested that IMF content is mainly determined by extra-myocellular lipids. At mRNA level, PLIN2 expression was higher in high IMF muscles (P<0.05). The results indicate for the first time that in pig muscle PLIN1 and PLIN2 proteins are localized in correspondence with extra and intra-myocellular lipids, respectively.

Association of PPARGC1A and CAPNS1 gene polymorphisms and expression with meat quality traits in pigs.

This study aimed to investigate the genes PPARGC1A (peroxisome proliferator-activated receptor gamma-coactivator 1A) and CAPNS1 (calpain small subunit 1) as candidate genes affecting meat quality traits in pigs. Four polymorphisms were identified in PPARCG1A and three in CAPNS1. The PPARGC1A polymorphism c.1288T>A was associated with pH and cooking loss in a F2 Duroc×Pietrain experimental cross (DuPi, n=313) and with pH values in Italian Large White (ILW, n=380) and Italian Landrace (ILA, n=158) populations (P<0.05). The CAPNS1 polymorphism c.429A>C was associated with pH and conductivity in DuPi and with meat color in ILA (P<0.05). PPARGC1A mRNA expression associated with drip loss (P<0.01) and the same tendency was found for CAPNS1 (P=0.06). The promoter methylation profiling suggested that methylation is not involved in CAPNS1 expression regulation. In conclusion, porcine PPARGC1A and CAPNS1 genes may affect meat quality traits, with breed-specific differences, and they could be used as markers for the improvement of meat quality in pigs.