Significance of Familial History of Prostate Cancer to Traditional Prognostic Variables, Genetic Biomarkers, and Recurrence after Radical Prostatectomy

Objectives. Prostate cancer (PCa) has a familial predisposition imparting an increased risk of developing the disease in those with a family history. The pathologic characteristics are similar to sporadic cases; however, the disease-free survival rates of hereditary PCa have recently been disputed, with one major study suggesting that familial cases have higher recurrence rates. Our study seeks to support or refute this association and to evaluate the genetic biomarkers p53, bcl-2, Ki-67, and neovascularity between familial and sporadic disease.Methods. We retrospectively reviewed data of 573 patients who underwent radical prostatectomy over an 11-year period. Of these, 474 patients had known family history data. Univariable statistical analysis using the Pearson chi-square test and Kaplan-Meier disease-free survival analysis was performed to identify any correlation between the tested variables and family history. Smaller subsets of this cohort that had available archival material for immunohistochemical staining and family history data were analyzed in a similar manner.Results. The preoperative variables (prostate-specific antigen, prostatic acid phosphatase, clinical stage, highest biopsy Gleason sum, and glandular differentiation) and postoperative variables (stage, highest Gleason sum, and glandular differentiation) did not correlate with family history. Kaplan-Meier disease-free survival analysis revealed no differences between sporadic and familial cases. The analysis of p53, bcl-2, Ki-67, and angiogenesis revealed that only increasing p53 expression and positive family history of PCa approached significance (P = 0.057).Conclusions. Prognostic variables routinely used in PCa and selected genetic biomarker immunostaining abnormalities are not significantly different in men with and without a family history of PCa. Disease-free survival after radical prostatectomy is also unaffected by family history.     

PharmAdhere: training German community pharmacists with Objective Structured Clinical Examinations

International Journal of Clinical Pharmacy - Background Pharmacists who engage in Pharmaceutical Care need skills to optimise responsible medication use and increase medication adherence....     

Identification of signature volatiles to discriminate Candida albicans,glabrata, krusei and tropicalis using gas chromatography and mass spectrometry

Oral candidiasis is the most frequent fungal infection of the oral cavity. Clinical diagnoses require mycological confirmation, which is time‐consuming in case of culture testing. The aim of the study was to identify signature volatiles to develop a chairside breath test to diagnose oral candidiasis. Headspaces above Candida albicans, glabrata, tropicalis, krusei cultures, and growth media as control were analysed after eight and 24 h using offline gas chromatography and mass spectrometry. The identification of signature volatiles was assisted using various microbial databases. Retrieved volatile patterns enabled Candida species discrimination in vitro. For C. albicans 3‐methyl‐2‐butanone and styrene and for C. krusei a combination of p‐xylene, 2‐octanone, 2‐heptanone and n‐butyl acetate were found to be specific. 1‐hexanol was found in C. tropicalis, but is emitted by a variety of other microorganisms. C. glabrata was characterised through the absence of these volatiles. The development of a breath test is a promising approach in confirming suspicions of oral candidiasis. To confirm the retrieved results in vivo, breath tests in affected and healthy subjects have to be performed.     

Triggering of NF‐κB in cytokine‐matured human DCs generates superior DCs for T‐cell priming in cancer immunotherapy

Understanding the signaling that governs the immunogenicity of human dendritic cells (DCs) is a prerequisite for improving DC‐based therapeutic vaccination strategies, in which the ability of DCs to induce robust and lasting Ag‐specific CTL responses is of critical importance. Cytokine‐matured DCs are regularly used, but to induce memory‐type CTLs, they require additional activation stimuli, such as CD4⁺ T‐cell help or TLR activation. One common denominator of these stimuli is the activation of NF‐κB. Here, we show that human monocyte‐derived, cytokine cocktail‐matured DCs transfected with constitutively active mutants of IκB kinases (caIKKs) by mRNA electroporation, further upregulated maturation markers, and secreted enhanced amounts of cytokines, including IL‐12p70, which was produced for more than 48 h after transfection. Most importantly, cytotoxic T cells induced by caIKK‐transfected DCs combined high CD27 expression, indicating a more memory‐like phenotype, and a markedly enhanced secondary expandability with a high lytic capacity. In contrast, CTLs primed and expanded with unmodified cytokine cocktail‐matured DCs did not maintain their proliferative capacity upon repetitive stimulations. We hypothesize that “designer” DCs expressing constitutively active IκB kinases will prove highly immunogenic also in vivo and possibly emerge as a new strategy to improve the clinical efficacy of therapeutic vaccinations against cancer and other chronic diseases.     

Nep1-like proteins from three kingdoms of life act as a microbe-associated molecular pattern in Arabidopsis

Necrosis and ethylene-inducing peptide 1 (Nep1)-like proteins (NLPs) are secreted by a wide range of plant-associated microorganisms. They are best known for their cytotoxicity in dicot plants that leads to the induction of rapid tissue necrosis and plant immune responses. The biotrophic downy mildew pathogen Hyaloperonospora arabidopsidis encodes 10 different noncytotoxic NLPs (HaNLPs) that do not cause necrosis. We discovered that these noncytotoxic NLPs, however, act as potent activators of the plant immune system in Arabidopsis thaliana. Ectopic expression of HaNLP3 in Arabidopsis triggered resistance to H. arabidopsidis, activated the expression of a large set of defense-related genes, and caused a reduction of plant growth that is typically associated with strongly enhanced immunity. N- and C-terminal deletions of HaNLP3, as well as amino acid substitutions, pinpointed to a small central region of the protein that is required to trigger immunity, indicating the protein acts as a microbe-associated molecular pattern (MAMP). This was confirmed in experiments with a synthetic peptide of 24 aa, derived from the central part of HaNLP3 and corresponding to a conserved region in type 1 NLPs that induces ethylene production, a well-known MAMP response. Strikingly, corresponding 24-aa peptides of fungal and bacterial type 1 NLPs were also able to trigger immunity in Arabidopsis. The widespread phylogenetic distribution of type 1 NLPs makes this protein family (to our knowledge) the first proteinaceous MAMP identified in three different kingdoms of life.Immune responses in plants generally start by receptor-mediated detection of nonself molecules that are conserved among different classes of microbes, both beneficial and pathogenic (1). These molecules often have essential functions in microbial fitness (2) and are known as microbe-associated molecular patterns (MAMPs). Upon their perception by the plant, MAMPs trigger basal immune responses (3), e.g., ethylene biosynthesis, production of reactive oxygen species, release of antimicrobial compounds (4), and in certain cases programmed cell death (2). Collectively, these responses contribute to resistance against nonadapted pathogens [MAMP-triggered immunity (MTI)].MAMPs of plant-infecting microbes have been described for bacteria, fungi, and oomycetes. Three characterized bacterial MAMPs are flagellin (5), EF-Tu (6), and peptidoglycan (7). Flagellin is the main protein of the bacterial flagellum, which is used by eubacteria for movement. A highly conserved fragment of 22 aa, named flg22 (5), is sufficient to activate MTI in Arabidopsis and other plant species. Elongation factor thermo unstable (EF-Tu) is an abundant and conserved bacterial protein that plays a central role in the elongation phase of protein synthesis. An 18-aa domain of EF-Tu, named elf18, is recognized as a MAMP in Brassicaceae species, but not in other tested plant families (6). Peptidoglycan (PGN), the third characterized bacterial MAMP, is a major structural component of most bacterial cell walls. PGN, consisting of strands of alternating N-acetylglucosamine and N-acetylmuramic acid residues, triggers immunity in Arabidopsis (7). An important fungal MAMP is chitin, a structural component of all fungal cell walls. Plants are able to recognize chitin, and fragments of 4–10 N-acetylglucosamine residues are the most potent inducers of defense (8). Recently, a second fungal MAMP was identified, a secreted polygalacturonase of Botrytis cinerea that triggers immunity in Arabidopsis (9).Four oomycete-derived MAMPs have been identified to date (10): (i) heptaglucoside fragments, originating from branched β-glucans that are major cell wall polysaccharides, and that trigger defense responses in many Fabaceous plants (11); (ii) glycoprotein 42, a calcium-dependent transglutaminase that functions in irreversible protein cross-linking and is abundant in Phytophthora cell walls, and a 13-aa peptide fragment thereof that elicit MTI responses in parsley (12) and potato (13); (iii) elicitins, secreted proteins with sterol-binding activity (14), which provoke necrosis in Nicotiana plants through induction of cell death (15); and (iv) the Phytophthora cellulose-binding elicitor lectin, which is thought to cause perturbation of the cell wall cellulose status, thereby triggering necrosis and MTI in tobacco and Arabidopsis (16, 17). Other groups of cell death-inducing proteins may also qualify as MAMPs based on their widespread occurrence among different pathogens (2), e.g., the Crinklers and the cytotoxic necrosis and ethylene-inducing peptide 1 (Nep1)-like proteins (NLPs) (10).Two major NLP types are found in bacteria, fungi, and oomycetes (18, 19) and are known to cause rapid necrosis and ethylene production in many dicot, but not in monocot plant species (18, 20). Type 2 NLPs differ from type 1 by an additional conserved second cysteine bridge and putative calcium-binding domain (19). In Arabidopsis, cytotoxic NLPs were found to activate immunity-related gene expression, which strongly overlapped with that induced by flg22 (21, 22). However, it was suggested that immune responses resulted from cytotoxicity. Moreover, necrosis was only induced upon treatment with the complete NLP protein (23). In vitro, cytotoxic NLPs cause rapid leakage of dicot membrane-derived vesicles, suggesting a direct cytolytic activity (24). The immunogenic effect of NLPs was therefore suggested to result from direct cellular damage (24), or release of damage-associated molecular patterns (3).Several plant-infecting oomycetes have large expansions of NLPs in their genomes (25–27), suggesting that these proteins play an important role in the pathogen’s lifestyle. A clear virulence function was observed for NLP_Pcc of the rot bacterium Pectobacterium carotovorum (27). Also, individual deletion of two NLP genes in the fungus Verticillium dahliae resulted in reduced virulence on different host plants (28). Five other NLP genes in this fungus encode noncytotoxic proteins (29), a phenomenon that is also observed in oomycetes. When tested by transient expression in tobacco, necrosis was only induced by 1 out of 3 tested NLPs of Phytophthora infestans (30), 8 out of 33 NLPs of Phytophthora sojae (31), whereas not a single 1 of 10 NLPs of Hyaloperonospora arabidopsidis tested caused necrosis (26). In contrast to cytotoxic NLPs that are mainly expressed during necrotrophic stages of infection, noncytotoxic NLPs appear to be expressed early during infection (26, 30), suggesting they serve an, as-yet-unknown, function during penetration or initial colonization of the host.In our search for the biological function of noncytotoxic NLPs of H. arabidopsidis, transgenic HaNLP-expressing Arabidopsis plants were generated that were severely stunted. In this paper, we show that Arabidopsis responds to noncytotoxic HaNLPs and small peptide fragments thereof that are highly conserved in type 1 NLPs. The peptides activate ethylene production and other typical MAMP-triggered defense responses, but not tissue necrosis, indicating they act as a MAMP. NLPs are not restricted to a single class of microbes but present in a broad range of mostly plant-associated microbes (bacteria, fungi, and oomycetes) belonging to three kingdoms of life, making this a MAMP with an unprecedented broad taxonomic occurrence.     

Prevalence of Sexual Aggression Victimization and Perpetration in a German University Student Sample

Archives of Sexual Behavior - This study examined the prevalence of sexual aggression perpetration and victimization in a sample of 1,172 students (755 female, 417 male) from four universities in...