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To test the hypothesis that genital skin and male urethra affected by lichen sclerosus (LS) has increased collagen content and altered collagen structure.We used picrosirius red to stain and image collagen in human urethral, vulvar, and foreskin specimens with and without LS. Using Image J software, we quantified and compared 1) collagen content (using two metrics: collagen proportionate area [CPA] and collagen fiber count), 2) collagen fiber length and width, and 3) collagen structure using the texture analysis technique gray level co-localization matrix (GLCM) with respect to LS status and tissue type.We analyzed 23 LS specimens (vulva n=9, urethra n=7, foreskin n=7) and 29 non-LS specimens (vulva n=9, urethra n=7, foreskin n=13). Fiber count and CPA were significantly higher in all LS specimens compared to non-LS specimens (CPA: mean±SD 0.971±0.03 vs 0.948±0.02, p0.007; fiber count: mean±SD = 2906±127 vs 2509±78 fibers; p = 0.003). Collagen fiber width and length were similar with respect to LS status. GLCM analysis showed decreased inverse difference moment and increased entropy in LS tissues indicative of less homogeneous and more disorganized tissue structure (p0.001).LS tissues have greater collagen content compared to non-LS tissues. Quantitative assessment of collagen organization, using GLCM, revealed less homogeneity and more disorganization of collagen in LS compared to non-LS tissues. Taken together, our findings suggest that alterations in physical tissue properties seen in LS may be due to both increased collagen abundance and altered structure.

Systemic sclerosis (SSc) is a clinically heterogeneous fibrotic disease with no effective treatment. Myofibroblasts are responsible for unresolving synchronous skin and internal organ fibrosis in SSc, but the drivers of sustained myofibroblast activation remain poorly understood. Using unbiased transcriptome analysis of skin biopsies, we identified the downregulation of SPAG17 in multiple independent cohorts of patients with SSc, and by orthogonal approaches, we observed a significant negative correlation between SPAG17 and fibrotic gene expression. Fibroblasts and endothelial cells explanted from SSc skin biopsies showed reduced chromatin accessibility at the SPAG17 locus. Remarkably, mice lacking Spag17 showed spontaneous skin fibrosis with increased dermal thickness, collagen deposition and stiffness, and altered collagen fiber alignment. Knockdown of SPAG17 in human and mouse fibroblasts and microvascular endothelial cells was accompanied by spontaneous myofibroblast transformation and markedly heightened sensitivity to profibrotic stimuli. These responses were accompanied by constitutive TGF-β pathway activation. Thus, we discovered impaired expression of SPAG17 in SSc and identified, to our knowledge, a previously unreported cell-intrinsic role for SPAG17 in the negative regulation of fibrotic responses. These findings shed fresh light on the pathogenesis of SSc and may inform the search for innovative therapies for SSc and other fibrotic conditions through SPAG17 signaling.

The invasive sea lamprey was a significant factor in the collapse of fish stocks in the Great Lakes, and it continues to threaten the multi-billion-dollar fishing industry. Thus, substantial resources are invested annually on sea lamprey control. Current control strategies have reduced sea lamprey populations by up to 90%, but they are expensive and have some limitations, e.g., lamprey-specific biocides applied to larval habitat impact native lampreys, and physical barriers that block adult lamprey access to spawning habitat impede migration of other fishes. Therefore, genetic control options which offer a theoretically powerful and effective pest control tool are being explored, although they have uncertain sociopolitical support, especially given the need to protect sea lamprey in their native range in Atlantic drainages. Here, we present an overview of genetic approaches with potential for application to sea lamprey control in the Great Lakes. We classify these approaches into two major categories: self-limiting (heritable sex ratio ratchet, Trojan gene, split gene drive) and self-sustaining (gene drive-based sex ratio distortion, homing suppression gene drive, toxin-antidote gene drives, and modification-type gene drives to aid suppression). We describe the technical aspects, challenges, and potential application of each method, focusing on gene drives, a fast-evolving research area that was only a distant option for sea lamprey control in previous reviews. We conclude that, given the risk of undesired spread of deleterious alleles from the Great Lakes, self-limiting genetic control options and confined gene drives will likely be preferred over unconfined gene drive options for sea lamprey control.

Low-grade serous ovarian cancer represents a minority of ovarian cancers and has distinctive features from high grade epithelial ovarian cancer. While less aggressive, in advanced stage they can be poorly chemo-responsive and incur a treatment challenge. Next generation sequencing of tumors has allowed for the potential for targeted therapy in cancer treatment, which can allow for avoidance of traditional cytotoxic chemotherapy. We present a case of a 56 year old female with advanced recurrent low grade serous ovarian cancer found to have NRAS mutation who underwent targeted therapy with trametinib with immediate and sustained disease response. We review the response and toxicity experienced by the patient, as well as treatment for her toxicity., Highlights • Trametinib was effective in low grade ovarian cancer with NRAS mutation. • Next generation sequencing of tumors can provide new options. • Common side effects of trametinib can be managed with dose reduction.

Propionibacterium acnes is a skin commensal bacterium that contributes to the development of acne vulgaris and other infections. Recent work revealed that P. acnes clinical isolates can be classified into distinct phylotypes, several of which have associations with healthy skin or acne. We sought to determine if these phylotypes induce different immunological responses and express protein factors that may contribute to their disease associations. We found that acne-associated P. acnes phylotypes induced 2- to 3-fold higher levels of IFN-γ and IL-17 in peripheral blood mononuclear cells compared with healthy phylotypes. On the other hand, P. acnes phylotypes associated with healthy skin induced 2- to 4-fold higher levels of IL-10. Comparative proteomic analysis of P. acnes phylotypes revealed a differential expression of several proteins, including an adhesion protein that was expressed at least 10-fold higher in acne-associated phylotypes and a cell surface hydrolase expressed in all phylotypes except those associated with healthy skin. Taken together, our data provide insight into how specific P. acnes phylotypes influence immune responses and the pathogenesis of acne.

Patients with solid tumors are at greatest risk for dying from their cancers in the five years following diagnosis. For most malignancies, deaths from other chronic diseases begin to exceed those from cancer at some point. As little is known about the causes of death among long-term survivors of ovarian cancer, we examined causes of death by years from diagnosis.The Surveillance, Epidemiology, and End Results (SEER) database was used to identify women diagnosed with ovarian cancer between 1988 and 2012. We compared causes of death by stage, age, and interval time after diagnosis.A total of 67,385 women were identified. For stage I neoplasms, 13.6% (CI, 13.0-14.2%) died from ovarian cancer, 4.2% (CI, 3.8-4.5%) from cardiovascular disease, 3.6% (CI, 3.3-3.9%) from other causes and 2.6% (CI, 2.4-2.9%) from other tumors; ovarian cancer was the leading cause of death until 7 years after diagnosis after which time deaths are more frequently due to other causes. For those with stage III-IV tumors, 67.8% (CI, 67.3-68.2%) died from ovarian cancer, 2.8% (CI, 2.6-2.9%) from other causes, 2.3% (CI, 2.2-2.4%) from cardiovascular disease and 1.9% (CI, 1.7-2.0%) from other cancers; ovarian cancer was the most frequent cause of death in years 1-15 after which time deaths were more commonly due to other causes.The probability of dying from ovarian cancer decreases with time. Ovarian cancer remains the most common cause of death for 15 years after diagnosis in women with stage III-IV tumors.

Although antibiotics are a common treatment for acne, the difficulties inherent to effective antimicrobial penetration in sebum and selective antimicrobial action in the skin are compounded by increasing resistance of Propionibacterium acnes clinical isolates. To address these problems, we engineered Pentobra, a peptide–aminoglycoside molecule that has multiple mechanisms of antibacterial action and investigated whether it can be a potential candidate for the treatment of acne. Pentobra combines the potent ribosomal activity of aminoglycosides with the bacteria-selective membrane-permeabilizing abilities of antimicrobial peptides. Pentobra demonstrated potent and selective killing of P. acnes but not against human skin cells in vitro. In direct comparison, Pentobra demonstrated bactericidal activity and drastically outperformed free tobramycin (by 5–7 logs) against multiple P. acnes clinical strains. Moreover, electron microscopic studies showed that Pentobra had robust membrane activity, as treatment with Pentobra killed P. acnes cells and caused leakage of intracellular contents. Pentobra may also have potential anti-inflammatory effects as demonstrated by suppression of some P. acnes–induced chemokines. Importantly, the killing activity was maintained in sebaceous environments as Pentobra was bactericidal against clinical isolates in comedones extracts isolated from human donors. Our work demonstrates that equipping aminoglycosides with selective membrane activity is a viable approach for developing antibiotics against P. acnes that are effective in cutaneous environments.

Advances in nanotechnology have demonstrated potential application of nanoparticles (NPs) for effective and targeted drug delivery. Here we investigated the antimicrobial and immunological properties and the feasibility of using NPs to deliver antimicrobial agents to treat a cutaneous pathogen. NPs synthesized with chitosan and alginate demonstrated a direct antimicrobial activity in vitro against Propionibacterium acnes, the bacterium linked to the pathogenesis of acne. By electron microscopy (EM) imaging, chitosan-alginate NPs were found to induce the disruption of the P. acnes cell membrane, providing a mechanism for the bactericidal effect. The chitosan-alginate NPs also exhibited anti-inflammatory properties as they inhibited P. acnes-induced inflammatory cytokine production in human monocytes and keratinocytes. Furthermore, benzoyl peroxide (BP), a commonly used antiacne drug, was effectively encapsulated in the chitosan-alginate NPs and demonstrated superior antimicrobial activity against P. acnes compared with BP alone while demonstrating less toxicity to eukaryotic cells. Together, these data suggest the potential utility of topical delivery of chitosan-alginate NP-encapsulated drug therapy for the treatment of dermatologic conditions with infectious and inflammatory components.

ResumenObjetivoDescribir el estado actual de los conocimientos sobre el documento de voluntades anticipadas (DVA) de los profesionales de atención primaria.EmplazamientoSAP Mataró Maresme del Institut Català de la Salut.DiseñoEstudio descriptivo y transversal.ParticipantesProfesionales de los equipos de atención primaria.MedicionesSe diseñó un cuestionario de autorrespuesta individual y anónima. Se definió un patrón de respuesta y un patrón de respuesta mínima correcta para cada bloque. Se realizan 2 pruebas de comprensibilidad y validez de los conceptos, y una prueba piloto de factibilidad.Resultados principalesSobre una población diana de 475 individuos se obtienen 227 respuestas (47%), de las que 219 (46%) fueron válidas. Según los colectivos profesionales, el 59% son médicos, el 28% son pediatras, el 7% son odontólogos, el 59% son diplomados en enfermería, el 12% son auxiliares de enfermería, el 30% son trabajadores sociales y el 25% son de atención al usuario. Solo 4 profesionales tienen redactado su propio DVA. El porcentaje de aciertos por bloques es el siguiente: el 83,8% de definición, el 4,1% de aspectos legales, el 0,5% de procedimiento-registro, el 1,4% de contenidos y el 38,6% de aplicación.ConclusionesLos profesionales tienen un conocimiento general sobre qué son las voluntades anticipadas y el DVA, pero saben poco de la normativa, el contenido y el registro. No hay diferencias significativas entre los colectivos analizados. El cuestionario parece útil para evaluar los conocimientos sobre el DVA.AbstractObjectiveTo assess the current state of knowledge and attitudes on the advance directives (living wills) document of professionals working in primary.DesignDescriptive, cross-sectional study.ParticipantsPrimary care professionals.SettingCatalonia Health Institute, Maresme Province, Barcelona.MethodsAn anonymous self-administered questionnaire was prepared with a correct answer model and a minimum correct level for each block of questions, as well as two comprehensibility and validity tests and a pilot feasibility test.ResultsFrom a target population of 475 individuals, t 227 (47%) responses were received (59% of GPs, 28% of paediatricians, 7% of dentists, 59% of nurses, 12% of assistant clinics, 30% of social workers and 25% of administrative assistants). Four people had written their own advance directive document. The percentage of correct answers was: definition block 83.8%, legal aspects 4.1%, procedure-register 0.5%; content 1.4%, application 38.6%.ConclusionsPrimary care workers have a general knowledge on the advance directives and the advance directive document but not enough about the law, content and registers. There was no significant difference between professional groups. The questionnaire appears useful for assessing knowledge on advance directives document.

The biogenesis of the bacterial flagellum and chemotaxis apparatus in both Escherichia coli and Caulobacter crescentus requires the ordered expression of over 40 genes whose expression is controlled by a trans-acting regulatory hierarchy. In C. crescentus, additional control mechanisms ensure that the transcription of these genes is initiated at the correct time in the cell cycle. We demonstrate here that two flagellar genes, flaE and fla Y, whose products function in trans to modulate the level of transcription of other flagellar genes, are themselves temporally controlled. DNA sequence analysis of the 3413 base-pairs encompassing the flaE and fla Y coding sequences and the 5′ regulatory region showed that flaE encodes a protein of 16,000 Mr and fla Y a, protein of 17,000 Mr. Evidence that flaE and fla Y are transcribed as a polycistronic message includes (1) the polar effect of Tn5 insertions; (2) deletion analysis showing that the flaE promoter is essential for complementation of both flaE and fla Y alleles; and (3) nuclease S1 assays showing protection of a transcript spanning both genes. The transcript start site in front of flaE was determined and the −10 region conforms to the E. coli sigma 28 promoter consensus sequence. Nuclease S1 analysis also revealed a protected fragment whose size was consistent with a transcript initiating in vivo at a consensus “nif” promoter sequence in front of the fla Y gene. The entire promoter region and an upstream consensus sequence that might be a regulatory element for the fla Y gene lies within the carboxyl-terminal coding sequence of the flaE gene.