A High-throughput Screening of a Chemical Compound Library in Ovarian Cancer Stem Cells.

BACKGROUND: Epithelial ovarian cancer has a poor prognosis, mostly due to its late diagnosis and the development of drug resistance after a first platinum-based regimen. The presence of a specific population of "cancer stem cells" could be responsible of the relapse of the tumor and the development of resistance to therapy. For this reason, it would be important to specifically target this subpopulation of tumor cells in order to increase the response to therapy. METHOD: We screened a chemical compound library assembled during the COST CM1106 action to search for compound classes active in targeting ovarian stem cells. We here report the results of the high-throughput screening assay in two ovarian cancer stem cells and the differentiated cells derived from them. RESULTS AND CONCLUSION: Interestingly, there were compounds active only on stem cells, only on differentiated cells, and compounds active on both cell populations. Even if these data need to be validated in ad hoc dose response cytotoxic experiments, the ongoing analysis of the compound structures will open up to mechanistic drug studies to select compounds able to improve the prognosis of ovarian cancer patients.

Ιnfluenza A(H3N2) genetic variants in vaccinated patients in northern Greece.

BACKGROUND: Influenza A(H3N2) viruses predominated during the influenza 2016/2017 season and showed extensive genetic diversification. A high vaccination failure rate was noticed during the 2016/17 season in Greece, especially among the elderly. OBJECTIVES: The scope of the study was to investigate the genetic characteristics of A(H3N2) circulating viruses and viruses detected in vaccinated patients. STUDY DESIGN: Virus samples originated from vaccinated and unvaccinated patients, obtained at the National Influenza Centre for northern Greece. Phylogenetic analysis and comparison of the haemagglutinin gene of the viruses to the vaccine virus A/Hong Kong/4801/2014 was performed. RESULTS: The majority of analysed viruses are clustering in the genetic clade 3C.2a, and in a newly emerged subclade, designated as 3C.2a1. The highest proportion of viruses detected in vaccinated patients fell into a distinct subcluster within the 3C.2a1 subclade, which is characterised by the amino acid substitutions N122D and T135K in haemagglutinin. CONCLUSIONS: Viruses that belong to the 3C.2a clade are generally considered to resemble antigenically to the northern hemisphere vaccine component A/Hong Kong/4801/2014 that was recommended by WHO to be included also into the 2017/18 vaccine. However, viruses belonging to a specific 3C.2a1 subcluster was extensively circulating in northern Greece and among vaccinated individuals. Both substitutions carried by this strain were located on antigenic sites and caused losses of N-linked glycosylation sites of the virus, which could potentially affect viral antigenicity. Further studies are needed to determine the antigenicity of this variant strain and its possible implication in vaccine effectiveness.

Pilot study of 19 patients with severe pemphigus: prophylactic treatment with rituximab does not appear to be beneficial.

AIMS: To assess the efficacy and safety of rituximab in refractory pemphigus and the possible benefit of an additional prophylactic infusion at 6 months. METHODS: Seventeen patients with pemphigus vulgaris, 1 with pemphigus foliaceus and 1 with pemphigus vegetans were treated with 4 weekly infusions of rituximab (375 mg/m(2)). Nine patients received an additional prophylactic infusion after 6 months while the rest received no maintenance therapy. In case of recurrence, an additional single infusion was administered. RESULTS: Control of the disease was obtained after 3-8 weeks. End of the consolidation phase for all patients was observed after 16 weeks. Patients remained in full remission for 7-42 months. All immunosuppressive agents, including prednisone, were discontinued after 2-12 months. The disease relapsed in 5 out of 9 patients who received the additional prophylactic infusion, and in 3 out of 10 patients among those skipping the prophylactic additional infusion. CONCLUSION: One course of rituximab and treatment of relapses is highly effective and well tolerated in the treatment of refractory pemphigus. In this pilot study of 19 patients, the prophylactic infusion does not appear to have provided any additional benefit to the patients receiving it.

Evaluation of the repellent effects of Nepeta parnassica extract, essential oil, and its major nepetalactone metabolite against mosquitoes.

The dichloromethane-methanol extract, the essential oil obtained by hydrodistillation from Nepeta parnassica, and the isolated 4aα,7α,7aß-nepetalactone were evaluated for their repellent effect against the mosquitoes Aedes (Stegomyia) cretinus Edwards and Culex pipiens pipiens biotype molestus Forskål. The chemical analysis of N. parnassica essential oil, dominated by oxygenated monoterpenes (87 %), revealed 4aα,7α,7aß-nepetalactone (36.8%), 1,8-cineole (25.5%), and 4aα,7ß,7aß-nepetalactone (11.1%) as the major constituents. The results of the insect bioassays showed that the essential oil and the dichloromethane-methanol extract of N. parnassica were very active against Aedes cretinus for up to 3 h and against Culex pipiens for up to 2 h post application. The isolated 4aα,7α,7aß-nepetalactone showed very high mosquito repellency for periods of at least 2 h against both species.

Development of a sensitive cost-effective capture ELISA for detection of murine monoclonal antibodies: correlation with SPR biosensor technology.

The development of antibodies for diagnostic and therapeutic applications in inflammatory diseases is a major focus for biotechnology and pharmaceutical companies. Production of monoclonal antibodies requires the development of fast, high-throughput methodologies for screening and selecting appropriate candidate antibodies for development. Capture (sandwich) enzyme linked immunosorbent assay (ELISA) provides a quick and reliable method that could be used for hybridoma screening of potential candidates accompanied with surface plasmon resonance (SPR) biosensor technology for identifying high affinity biomolecular interactions. A sensitive, cost-effective, robust and accurate capture ELISA for detection of murine monoclonal antibodies in culture supernatants was developed. This assay was optimized for high sensitivity and specificity with a capture anti-mouse polyclonal antibody. Using serial dilutions of a defined murine IgG antibody, a linear dose-response was observed between 2 and 1200 ng/ml antibody with a coefficient of determination r2 of 0.98. The detection limit of the assay was established as 2ng/ml (12.5pM). A similar concentration-dependent doseresponse was also observed using serial dilutions of antibody-containing supernatants from anti-alpha glycophorinproducing hybridomas (detection limit 1:2000). Specific capture of antibodies from supernatants in a similar setting was also confirmed using SPR biosensor technology and correlated well with the immunoassay results. The latter technology can be performed in order to provide quick screening results and kinetic analysis of antibody binding interactions aiming at identifying candidates with high affinity and specificity.

Comparison of two methods for identifying dietary patterns associated with obesity in preschool children: the GENESIS study.

BACKGROUND/OBJECTIVES: The aim of this work was to identify dietary patterns based on reduced rank regression (RRR) and principal component analysis (PCA) and to evaluate the association of these patterns with the prevalence of childhood obesity. SUBJECTS/METHODS: A sample of 2317 toddlers and preschoolers from Greece (Growth, Exercise and Nutrition Epidemiological Study In preSchoolers) was used. In total, 12 food groups were used as predictors of RRR and PCA. Nutrients such as total fat, simple carbohydrate and fiber intake were used as response variables to apply RRR. RESULTS: One factor/pattern was retained from RRR and PCA in order to ensure the comparability of the methods. The pattern derived from PCA was mainly characterized by consumption of fruits, vegetables, legumes, fish and seafood, grains and oils. This pattern explained 12.5% of the total variation in food groups. On the other hand, the pattern extracted from RRR was mainly characterized by reduced consumption of fruits, vegetables and legumes, and by increased consumption of sweets and red meat. The pattern derived from RRR explained 8.2% of the total variation in food groups. Simple and multiple logistic regression revealed that the pattern extracted from RRR is significantly associated with the prevalence of childhood obesity (OR=1.11, 95% CI: 1.00-1.28 for each unit increase of dietary pattern) as opposed to the pattern derived from PCA. CONCLUSIONS: The preferable technique to derive dietary patterns related to childhood obesity seems to be RRR compared with PCA.

Developmental changes in adiposity in toddlers and preschoolers in the GENESIS study and associations with the ACE I/D polymorphism.

OBJECTIVES: To investigate the relationship between the angiotensin I-converting enzyme 1 (ACE) I/D polymorphism and adiposity-related phenotypes in a large cohort of toddlers and preschoolers. METHODS: Body composition measurements and DNA samples were obtained from 2102 Greek children aged 1-6 years, as part of a large-scale epidemiological study (GENESIS). All children were genotyped for the ACE I/D polymorphism and gender- and age-stratified statistical analyses were performed. RESULTS: In girls aged 4-6 years, the D-allele was associated with higher measurements of body mass index (BMI) (P=0.018), waist (P=0.001) and upper arm (P=0.013) circumferences, genotype accounting for 2.5, 4 and 3% of the phenotypic variance, respectively. In boys, the D-allele showed strong associations with lower BMI (P=0.001) at the age of 1-2 years that explained 17% of the phenotypic variance and with larger suprailiac skinfold (P=0.008) at 3-4 years old that explained 2% of the variance. No other significant associations between the ACE I/D polymorphism and adiposity-related phenotypes were found. In girls, the age at which significant associations were revealed coincided with the age at which BMI was observed to increase after its developmental nadir, but this feature of the association was not observed in boys. CONCLUSIONS: The ACE I/D polymorphism is associated with developmental and physiological changes in adiposity-related traits during early childhood in a gender- and age-specific manner.

Chemical composition and in vitro antimicrobial activity of the essential oils of flower heads and leaves of Santolina rosmarinifolia L. from Romania.

The essential oils of the flower heads and leaves of Santolina rosmarinifolia L. (Compositae) cultivated in Romania were obtained through hydrodistillation and analysed by GC-MS. Forty-two components were identified, representing 92.3-94.0% of the whole composition of the samples. Their in vitro antimicrobial activity against gram-positive and gram-negative bacteria strains and the fungus Candida albicans was determined using the agar-diffusion method and the broth dilution method.

Seborrhoeic keratosis or occult malignant neoplasm of the skin?

BACKGROUND: Seborrhoeic keratosis is generally considered to be a benign lesion of the skin. OBSERVATION: We present the case of a 68-year-old male who presented with clinically typical seborrhoeic keratosis that later histological examination showed partially covered an occult basal cell carcinoma. OBJECTIVE: To have an indication of what percentage of clinically apparent seborrhoeic keratoses may be associated with some form of histologically proven skin malignancy. METHODS: We carried out a retrospective analysis of approximately 23,000 histopathological examinations done on specimens from dermatological lesions. RESULTS: Fifty-nine (11.9%) clinically apparent seborrhoeic keratoses were later histologically diagnosed as basal cell carcinomas, 17 (3.4%) as squamous cell carcinomas, and five (1.01%) as malignant melanomas. CONCLUSIONS: Although the association of seborrhoeic keratosis and skin malignancy appears to be relatively uncommon, the possibility of such an association cannot be ruled out.

Allergy to olive pollen: a study of four family members.

We describe four family members with respiratory and dermatological manifestations of olive pollen allergy. The purpose of this study was 1) to investigate whether these patients' sera react to the same or different olive allergens, and 2) to identify common HLA class II antigens.

A simple methodological approach for counting and identifying culturable viruses adsorbed to cellulose nitrate membrane filters.

We identified conditions under which Buffalo green monkey cells grew on the surfaces of cellulose nitrate membrane filters in such a way that they covered the entire surface of each filter and penetrated through the pores. When such conditions were used, poliovirus that had previously been adsorbed on the membranes infected the cells and replicated. A plaque assay method and a quantal method (most probable number of cytopathic units) were used to detect and count the viruses adsorbed on the membrane filters. Polioviruses in aqueous suspensions were then concentrated by adsorption to cellulose membrane filters and were subsequently counted without elution, a step which is necessary when the commonly used methods are employed. The pore size of the membrane filter, the sample contents, and the sample volume were optimized for tap water, seawater, and a 0.25 M glycine buffer solution. The numbers of viruses recovered under the optimized conditions were more than 50% greater than the numbers counted by the standard plaque assay. When ceftazidime was added to the assay medium in addition to the antibiotics which are typically used, the method could be used to study natural samples with low and intermediate levels of microbial pollution without decontamination of the samples. This methodological approach also allowed plaque hybridization either directly on cellulose nitrate membranes or on Hybond N+ membranes after the preparations were transferred.

Chemotherapy-induced Raynaud's phenomenon.

BACKGROUND: Various antineoplastic agents can cause Raynaud's phenomenon, as can malignant diseases themselves. OBJECTIVE: To review the clinical characteristics of chemotherapy-induced Raynaud's phenomenon and compare them with those of malignancy-associated Raynaud's phenomenon. SUMMARY: Chemotherapy-induced Raynaud's phenomenon most commonly occurs in patients with testicular cancer who receive bleomycin either as a single agent or as part of a multiple-drug chemotherapeutic regimen. It tends to resolve spontaneously, especially after discontinuation of the inducing antineoplastic agent, and rarely causes significant functional impairment. However, it tends to recur with subsequent administration of the drug. In contrast, malignancy-associated Raynaud's phenomenon is rarer, causes more severe symptoms, and usually occurs in older patients with more advanced cancer. CONCLUSIONS: As more patients with cancer undergo chemotherapy, physicians should be aware of the potential delayed toxic effects of antineoplastic drugs.