Molecular Characterization and Patient Outcome of Melanoma Nodal Metastases and an Unknown Primary Site

Background Melanoma of unknown primary site (MUP) is not a completely understood entity with nodal metastases as the most common first clinical manifestation. The aim of this multicentric study was to assess frequency and type of oncogenic BRAF/NRAS/KIT mutations in MUP with clinically detected nodal metastases in relation to clinicopathologic features and outcome. Materials and Methods We analyzed series of 103 MUP patients (period: 1992-2010) after therapeutic lymphadenectomy (LND): 40 axillary, 47 groin, 16 cervical, none treated with BRAF inhibitors. We performed molecular characterization of BRAF/NRAS/KIT mutational status in nodal metastases using direct sequencing of respective coding sequences. Median follow-up time was 53 months. Results BRAF mutations were detected in 55 cases (53 %) (51 V600E, 93 %; 4 others, 7 %), and mutually exclusive NRAS mutations were found in 14 cases (14 %) (7 p.Q61R, 4 p.Q61K, 2 p.Q61H, 1 p.Q13R). We have not detected any mutations in KIT. The 5-year overall survival (OS) was 34 %; median was 24 months. We have not found significant correlation between mutational status (BRAF/NRAS) and OS; however, for BRAF or NRAS mutated melanomas we observed significantly shorter disease-free survival (DFS) when compared with wild-type melanoma patients (p = .04; 5-year DFS, 18 vs 19 vs 31 %, respectively). The most important factor influencing OS was number of metastatic lymph nodes >1 (p = .03). Conclusions Our large study on molecular characterization of MUP with nodal metastases showed that MUPs had molecular features similar to sporadic non-chronic-sun-damaged melanomas. BRAF/NRAS mutational status had negative impact on DFS in this group of patients. These observations might have potential implication for molecular-targeted therapy in MUPs

The power of the Mediator complex-Expanding the genetic architecture and phenotypic spectrum of MED12 -related disorders

MED12 is a member of the large Mediator complex that controls cell growth, development, and differentiation. Mutations in MED12 disrupt neuronal gene expression and lead to at least three distinct X-linked intellectual disability (XLID) syndromes (FG, Lujan-Fryns, and Ohdo). Here we describe six families with missense variants in MED12 (p.(Arg815Gln), p.(Val954Gly), p.(Glu1091Lys),p.(Arg1295Cys), p.(Pro1371Ser) and p.(Arg1148His), the latter being firstly reported in affected females) associated with a continuum of symptoms rather than distinct syndromes. The variants expanded the genetic architecture and phenotypic spectrum of MED12-related disorders. New clinical symptoms included brachycephaly, anteverted nares, bulbous nasal tip, prognathism, deep set eyes, and single palmar crease. We showed that MED12 variants, initially implicated in X-linked recessive disorders in males, may predict a potential risk for phenotypic expression in females, with no correlation of the X chromosome inactivation pattern in blood cells. Molecular modeling (Yasara Structure) performed to model the functional effects of the variants strongly supported the pathogenic character of the variants examined. We demonstrated that molecular modeling is a useful method for in silico testing of potential functional effects of MED12 variants and thus can be a valuable addition to the interpretation of the clinical and genetic findings

Zalecenia Polskiej Grupy Mięsakowej w odniesieniu do postępowania diagnostyczno-terapeutycznego oraz kontroli u chorych na neurofibromatozę typu 1 (NF1) oraz związanego z nią złośliwego nowotworu osłonek nerwów obwodowych

Type 1 neurofibromatosis (NF1 syndrome in von Recklinghausen’s disease) is inherited as an autosomal dominant disease, caused by mutations in the NF1 gene encoding the neurofibromin protein. NF1 patients are at an increased risk of the develop­ment of a malignant neoplasm and their life span is shorter by 20 years than that of the general population. National Institute of Health (NIH) criteria make a diagnosis possible from about 4 years of age. Examination of children and adults should encom­pass a physical and a subjective component, but also next-generation sequencing (NGS) genetic analysis, histopathological examination of skin lesions, neurological, ophthalmological and radiological examination. If a malignant peripheral nerve sheath tumor (MNPST) is diagnosed in a patient with NF1, the therapeutic procedure should not differ from the general principles of treating soft tissue sarcomas. Patients from the high risk group should be monitored at least once a year, the remaining patients once every 2–3 years by a specialized medical team, and every year by their primary physicians, internal medicine specialists and dermatologists. Patients should have access to genetic counselling.Neurofibromatoza typu 1 (zespół NF1 w chorobie Recklinghausena, nerwiakowłókniakowatość typu 1), jest dziedziczona au­tosomalnie dominująco, a odpowiadają za nią mutacje genu NF1 kodującego białko neurofibrominy. Pacjenci z NF1 są naraże­ni na zwiększone ryzyko rozwoju nowotworu złośliwego i żyją około 20 lat krócej niż populacja ogólna. Kryteria National Insti­tute of Health (NIH) umożliwiają postawienie diagnozy już około 4 roku życia. Badanie dzieci i dorosłych powinno objąć bada­nie przedmiotowe i podmiotowe, ale też badanie genetyczne techniką sekwencjonowania nowej generacji (NGS), badanie histopatologiczne zmian skóry, badanie neurologiczne, okulistyczne i radiologiczne. W przypadku postawienia roz­poznania złośliwego nowotworu osłonek nerwów obwodowych (malignant peripheral nerve sheath tumor – MPNST) u chorego na NF1 postępowanie terapeutyczne nie powinno odbiegać od ogólnych zasad leczenia mięsaków tkanek miękkich. Pacjenci z grupy wysokiego ryzyka powinni być monitorowani przynajmniej raz w roku, pozostali – raz na 2–3 lata – przez zespół lekarzy specjalistów, a co roku przez lekarzy podstawowej opieki zdrowotnej (POZ), chorób wewnętrznych i dermatologów. Pacjentom należy zapewnić poradnictwo genetyczne

Hearing screening among first-grade children in rural areas and small towns in Małopolskie voivodeship, Poland

Undiagnosed hearing deficits hamper a child’s ability to learn. Hearing screening in school aged children helps detect educationally significant hearing loss and prevents negative impacts on academic achievement. The main purpose of this study was to improve early detection and assess the incidence of hearing disorders in first-graders from rural areas and small towns in the Małopolskie Voivodeship of Poland. There were 5029 children aged 6–7 years. Hearing thresholds were measured over the frequency range 0.5–8 kHz. A result was considered positive (abnormal) if the hearing threshold was worse than 20 dB HL at one or more frequencies. The prevalence of hearing loss was estimated in terms of four-frequency hearing loss, high-frequency hearing loss, and low-frequency hearing loss. Parents filled in a brief audiological questionnaire. The analysis was performed using IBM SPSS Statistics, version 24. Of all the children, 20.5% returned a positive result and were referred for further audiological diagnoses. The estimated prevalence of hearing loss was 11.6%, made up of 6.5% with FFHL, 7.6% with HFHL, and 8.2% with LFHL. This study showed that large numbers of children in the district had hearing problems. Adoption of hearing screening in primary schools is recommended as a routine procedure within preventive pediatric health care

A multi-proxy, diachronic and spatial perspective on the urban activities within an indigenous community in medieval Riga, Latvia

The research presented here represents thefirst urban medieval context in Latvia where an integrated, multi-proxy environmental sampling strategy has been applied. The establishment of Riga, the modern capital of Latvia, is synonymous with the Livonian Crusade, and the foundation of the medieval town is examined here. This study uses an intra-site comparison of environmental datasets from several buildings to provide a unique, high resolution, diachronic analysis of the daily life of the inhabitants within the pre-Hansa town, and specifically of the indigenous ‘Liv’ population during the period of the Livonian crusades, 1198e1291. The integrated zooarchaeological, archaebotanical, and geochemical datasets from two successive phases of buildings show a pattern of tradition and continuity in indigenous practices within the ‘Liv District’. Despite being located within Riga with access to wide trade networks, the environmental results from ‘Liv district’ show the self-contained, insular nature of the diet and craft activities of the inhabitants who exploited a range of open grassland, wetland and forest edge environments around the town

Pathogenic Mutations and Putative Phenotype-Affecting Variants in Polish Myofibrillar Myopathy Patients

Myofibrillar myopathies (MFM) are heterogeneous hereditary muscle diseases with characteristic myopathological features of Z-disk dissolution and aggregates of its degradation products. The onset and progression of the disease are variable, with an elusive genetic background, and around half of the cases lacking molecular diagnosis. Here, we attempted to establish possible genetic foundations of MFM by performing whole exome sequencing (WES) in eleven unrelated families of 13 patients clinically diagnosed as MFM spectrum. A filtering strategy aimed at identification of variants related to the disease was used and included integrative analysis of WES data and human phenotype ontology (HPO) terms, analysis of muscle-expressed genes, and analysis of the disease-associated interactome. Genetic diagnosis was possible in eight out of eleven cases. Putative causative mutations were found in the DES (two cases), CRYAB, TPM3, and SELENON (four cases) genes, the latter typically presenting with a rigid spine syndrome. Moreover, a variety of additional, possibly phenotype-affecting variants were found. These findings indicate a markedly heterogeneous genetic background of MFM and show the usefulness of next generation sequencing in the identification of disease-associated mutations. Finally, we discuss the emerging concept of variant load as the basis of phenotypic heterogeneity

Contribution of RIT1 mutations to the pathogenesis of Noonan syndrome: four new cases and further evidence of heterogeneity.

Noonan syndrome (NS) is a common developmental disorder presenting with dysmorphic craniofacial features, heart defects, and short stature. It belongs to the group of RASopathies caused by germline mutations in genes encoding proteins involved in the RAS/MAPK signaling pathway. Although mutations in nine genes are known to cause NS, approximately 30% of the cases still have unexplained etiology. To identify the new causative genes, 42 patients with a clinical diagnosis of NS, who had negative results on Sanger sequencing of PTPN11, SOS1, and RAF1 (the most common NS genes), were selected for whole exome sequencing. In two patients, mutations in recently described new NS gene—RIT1 were found (c.244T>G [p.Phe82Val] and c.270G>C [p.Met90Ile]). Further analysis of a larger cohort (n = 64) of NS patients with classic Sanger sequencing revealed the presence of RIT1 mutation c.284G>C (p.Gly95Ala) in two additional patients. All the detected mutations were localized in switch II domain responsible for GTPase activity. The modeling of RIT1 protein structure revealed that the mutated amino acids and their interacting residues are evolutionary conserved and any residue replacement might change the structural stability and/or protein internal dynamics influencing catalytic activity of the protein. It seems that the identified mutations might alter protein function and therefore, the activity of ERK and P38 MAPK pathways, thus underlying the specific phenotype observed in NS patients. Our study independently confirms the role of RIT1 in the pathogenesis of Noonan syndrome

Structural patterns enhancing the antibacterial activity of metallacarborane-based antibiotics

Healthcare systems heavily rely on antibiotics to treat bacterial infections but widespread of the multidrug-resistant bacteria puts this strategy in danger. Novel drugs capable of overcoming current resistances are needed if our ability to treat bacterial infections is to be maintained. Boron clusters offer a valuable possibility to create a new class of antibiotics and expand antibiotic’s chemical space beyond conventional carbon-based molecules. In this work, we identified the two promising structural patterns providing cobalta bis(dicarbollide)(COSAN)-based compounds with potent and selective activity toward Staphylococcus aureus (including clinical strains): introduction of the α-amino acid amide and addition of iodine directly to the metallacarborane cage. Furthermore, we found that proper hydrophilic-lipophilic balance is crucial for the selective activity of the tested compounds toward S. aureus over mammalian cells. The patterns proposed in this paper can be useful in the development of metallacarborane-based antibiotics with potent antibacterial properties and low cytotoxicity