Oncological and functional outcome of conservative surgery for primary supraglottic cancer

The aim of this study was to verify the oncological and functional outcome of conservative surgical treatment of primary supraglottic squamous cell carcinoma (SGSCC) and related neck disease in order to verify the effectiveness of supraglottic laryngectomy (SL) and the validity of an "observation" policy in the control of clinically negative (NO) necks. Of a total of 252 consecutive patients affected by primary SGSCC seen between 1975 and 1990 at the Department of Otolaryngology of the University of Perugia (1975-1987) and the Catholic University of the Sacred Heart of Rome (1988-1990), a subset of 132 patients treated with classical SL was evaluated after presenting sufficient clinicopathological data and a follow-up period of at least 5 years. Tumors were staged according to the 1992 UICC TNM classification and grouped into stages I-II (n = 94) and III-IV (n = 38). Comprehensive neck dissections were performed only in the clinically positive (N+) necks (25/132 cases), while in the clinically NO ones (107/132 cases) an "observation" policy under strict follow-up conditions was adopted. After primary surgery, the 5-year relapse-free survival (RFS) was 74%. The RFS was 80% for T1-2 disease and 65% for T3. The RFS was 80% for stages I-II tumors and 71% for stages III-IV. The actual 5-year overall survival (OS) was 89% for T1-T2 tumors and 67% for T3 disease or 93% for stages I-II and 69% for stages III-IV. The OS was 89% for NO neck and 73% for N+. The 5-year-metastasis-free survival (MFS) was 83% for NO patients, 74% for N+, 84% for T1-T2 NO, 71% for T1-T2 N+, 81% for T3 NO and 68% for T3 N+. In all, SL was found to be highly effective in the management of primary SGSCC. In the presence of clinically NO neck "observation" under strict follow-up with therapeutic comprehensive neck dissection for delayed nodal recurrence, SL was suitable for controlling the neck cancer, as well as for salvaging recurrent disease. Bilateral elective, selective or functional neck dissection in every instance of supraglottic cancer was best performed only in those SGSCC patients who were more likely to have occult nodal disease on the basis of biological factors and imaging data

About the parabolic relation existing between the skewness and the kurtosis in time series of experimental data

In this work we investigate the origin of the parabolic relation between skewness and kurtosis often encountered in the analysis of experimental time-series. We argue that the numerical values of the coefficients of the curve may provide informations about the specific physics of the system studied, whereas the analytical curve per se is a fairly general consequence of a few constraints expected to hold for most systems.Comment: To appear in Physica Script

A note on abscissas of Dirichlet series

[EN] We present an abstract approach to the abscissas of convergence of vector-valued Dirichlet series. As a consequence we deduce that the abscissas for Hardy spaces of Dirichlet series are all equal. We also introduce and study weak versions of the abscissas for scalar-valued Dirichlet series.A. Defant: Partially supported by MINECO and FEDER MTM2017-83262-C2-1-P. A. Pérez: Supported by La Caixa Foundation, MINECO and FEDER MTM2014-57838-C2-1-P and Fundación Séneca - Región de Murcia (CARM 19368/PI/14). P. Sevilla-Peris: Supported by MINECO and FEDER MTM2017-83262-C2-1-P.Defant, A.; Pérez, A.; Sevilla Peris, P. (2019). A note on abscissas of Dirichlet series. Revista de la Real Academia de Ciencias Exactas Físicas y Naturales Serie A Matemáticas. 113(3):2639-2653. https://doi.org/10.1007/s13398-019-00647-yS263926531133Bayart, F.: Hardy spaces of Dirichlet series and their composition operators. Mon. Math. 136(3), 203–236 (2002)Bohnenblust, H.F., Hille, E.: On the absolute convergence of Dirichlet series. Ann Math. 32(3), 600–622 (1931)Bohr, H.: Über die Bedeutung der Potenzreihen unendlich vieler Variablen in der Theorie der Dirichlet–schen Reihen $$\sum \,\frac{a_n}{n^s}$$ ∑ a n n s . Nachr. Ges. Wiss. Göttingen, Math. Phys. Kl., pp. 441–488 (1913)Bonet, J.: Abscissas of weak convergence of vector valued Dirichlet series. J. Funct. Anal. 269(12), 3914–3927 (2015)Carando, D., Defant, A., Sevilla-Peris, P.: Bohr’s absolute convergence problem for $$\cal{H}_p$$ H p -Dirichlet series in Banach spaces. Anal. PDE 7(2), 513–527 (2014)Carando, D., Defant, A., Sevilla-Peris, P.: Some polynomial versions of cotype and applications. J. Funct. Anal. 270(1), 68–87 (2016)Defant, A., García, D., Maestre, M., Pérez-García, D.: Bohr’s strip for vector valued Dirichlet series. Math. Ann. 342(3), 533–555 (2008)Defant, A., García, D., Maestre, M., Sevilla–Peris, P.: Dirichlet Series and Holomorphic Funcions in High Dimensions, vol. 37 of New Mathematical Monographs. Cambridge University Press, Cambridge (2019)Defant, A., Pérez, A.: Optimal comparison of the $$p$$ p -norms of Dirichlet polynomials. Israel J. Math. 221(2), 837–852 (2017)Defant, A., Pérez, A.: Hardy spaces of vector-valued Dirichlet series. Studia Math. 243(1), 53–78 (2018)Diestel, J., Jarchow, H., Tonge, A.: Absolutely summing operators, vol. 43 of Cambridge studies in advanced mathematics. Cambridge University Press, Cambridge (1995)Maurizi, B., Queffélec, H.: Some remarks on the algebra of bounded Dirichlet series. J. Fourier Anal. Appl. 16(5), 676–692 (2010)Queffélec, H., Queffélec, M.: Diophantine approximation and Dirichlet series, vol. 2 of Harish–Chandra research institute lecture notes. Hindustan Book Agency, New Delhi (2013

CRISPR/Cas9 DNA cleavage at SNP-derived PAM enables both in vitro and in vivo KRT12 mutation-specific targeting

CRISPR/Cas9-based therapeutics hold the possibility for permanent treatment of genetic disease. The potency and specificity of this system has been used to target dominantly inherited conditions caused by heterozygous missense mutations through inclusion of the mutated base in the short-guide RNA (sgRNA) sequence. This research evaluates a novel approach for targeting heterozygous single-nucleotide polymorphisms (SNPs) using CRISPR/Cas9. We determined that a mutation within KRT12, which causes Meesmann's epithelial corneal dystrophy (MECD), leads to the occurrence of a novel protospacer adjacent motif (PAM). We designed an sgRNA complementary to the sequence adjacent to this SNP-derived PAM and evaluated its potency and allele specificity both in vitro and in vivo. This sgRNA was found to be highly effective at reducing the expression of mutant KRT12 mRNA and protein in vitro. To assess its activity in vivo we injected a combined Cas9/sgRNA expression construct into the corneal stroma of a humanized MECD mouse model. Sequence analysis of corneal genomic DNA revealed non-homologous end-joining repair resulting in frame-shifting deletions within the mutant KRT12 allele. This study is the first to demonstrate in vivo gene editing of a heterozygous disease-causing SNP that results in a novel PAM, further highlighting the potential for CRISPR/Cas9-based therapeutics

A novel role for CRIM1 in the corneal response to UV and pterygium development

Pterygium is a pathological proliferative condition of the ocular surface, characterised by formation of a highly vascularised, fibrous tissue arising from the limbus that invades the central cornea leading to visual disturbance and, if untreated, blindness. Whilst chronic ultraviolet (UV) light exposure plays a major role in its pathogenesis, higher susceptibility to pterygium is observed in some families, suggesting a genetic component. In this study, a Northern Irish family affected by pterygium but reporting little direct exposure to UV was identified carrying a missense variant in CRIM1 NM_016441.2: c.1235 A > C (H412P) through whole-exome sequencing and subsequent analysis. CRIM1 is expressed in the developing eye, adult cornea and conjunctiva, having a role in cell differentiation and migration but also in angiogenesis, all processes involved in pterygium formation. We demonstrate elevated CRIM1 expression in pterygium tissue from additional individual Northern Irish patients compared to unaffected conjunctival controls. UV irradiation of HCE-S cells resulted in an increase in ERK phosphorylation and CRIM1 expression, the latter further elevated by the addition of the MEK1/2 inhibitor, U0126. Conversely, siRNA knockdown of CRIM1 led to decreased UV-induced ERK phosphorylation and increased BCL2 expression. Transient expression of the mutant H412P CRIM1 in corneal epithelial HCE-S cells showed that, unlike wild-type CRIM1, it was unable to reduce the cell proliferation, increased ERK phosphorylation and apoptosis induced through a decrease of BCL2 expression levels. We propose here a series of intracellular events where CRIM1 regulation of the ERK pathway prevents UV-induced cell proliferation and may play an important role in the in the pathogenesis of pterygium

Nanoemulsions of Satureja montana essential oil. Antimicrobial and antibiofilm activity against avian Escherichia coli strains

Satureja montana essential oil (SEO) presents a wide range of biological activities due to its high content of active phytochemicals. In order to improve the essential oil’s (EO) properties, oil in water nanoemulsions (NEs) composed of SEO and Tween-80 were prepared, characterized, and their antimicrobial and antibiofilm properties assayed against Escherichia coli strains isolated from healthy chicken. Since surfactant and oil composition can strongly influence NE features and their application field, a ternary phase diagram was constructed and evaluated to select a suitable sur-factant/oil/water ratio. Minimal inhibitory concentration and minimal bactericidal concentration of NEs, evaluated by the microdilution method, showed that the SEO NE formulation exhibited higher inhibitory effects against planktonic E. coli than SEO alone. The quantification of biofilm production in the presence of NEs, assessed by crystal violet staining and scanning electron microscopy, evi-denced that sub-MIC concentrations of SEO NEs enable an efficient reduction of biofilm production by the strong producer strains. The optimized nanoemulsion formulation could ensure food safety quality, and counteract the antibiotic resistance of poultry associated E. coli, if applied/aerosolized in poultry farms

Keratin 12 missense mutation induces the unfolded protein response and apoptosis in meesmann epithelial corneal dystrophy

Meesmann epithelial corneal dystrophy (MECD) is a rare autosomal dominant disorder caused by dominant-negative mutations within the KRT3 or KRT12 genes, which encode the cytoskeletal protein keratins K3 and K12, respectively. To investigate the pathomechanism of this disease, we generated and phenotypically characterized a novel knock-in humanized mouse model carrying the severe, MECD-associated, K12-Leu132Pro mutation. Although no overt changes in corneal opacity were detected by slit-lamp examination, the corneas of homozygous mutant mice exhibited histological and ultrastructural epithelial cell fragility phenotypes. An altered keratin expression profile was observed in the cornea of mutant mice, confirmed by western blot, RNA-seq and quantitative real-time polymerase chain reaction. Mass spectrometry (MS) and immunohistochemistry demonstrated a similarly altered keratin profile in corneal tissue from a K12-Leu132Pro MECD patient. The K12-Leu132Pro mutation results in cytoplasmic keratin aggregates. RNA-seq analysis revealed increased chaperone gene expression, and apoptotic unfolded protein response (UPR) markers, CHOP and Caspase 12, were also increased in the MECD mice. Corneal epithelial cell apoptosis was increased 17-fold in the mutant cornea, compared with the wild-type (P < 0.001). This elevation of UPR marker expression was also observed in the human MECD cornea. This is the first reporting of a mouse model for MECD that recapitulates the human disease and is a valuable resource in understanding the pathomechanism of the disease. Although the most severe phenotype is observed in the homozygous mice, this model will still provide a test-bed for therapies not only for corneal dystrophies but also for other keratinopathies caused by similar mutations