Extra-virgin olive oil phenols block cell cycle progression and modulate chemotherapeutic toxicity in bladder cancer cells

Epidemiological data indicate that the daily consumption of extra‑virgin olive oil (EVOO), a common dietary habit of the Mediterranean area, lowers the incidence of certain types of cancer, in particular bladder neoplasm. The aim of the present study was to evaluate the antiproliferative activity of polyphenols extracted from EVOO on bladder cancer (BCa), and to clarify the biological mechanisms that trigger cell death. Furthermore, we also evaluated the ability of low doses of extra‑virgin olive oil extract (EVOOE) to modulate the in vitro activity of paclitaxel or mitomycin, two antineoplastic drugs used in the management of different types of cancer. Our results showed that EVOOE significantly inhibited the proliferation and clonogenic ability of T24 and 5637 BCa cells in a dose‑dependent manner. Furthermore, cell cycle analysis after EVOOE treatment showed a marked growth arrest prior to mitosis in the G2/M phase for both cell lines, with the subsequent induction of apoptosis only in the T24 cells. Notably, simultaneous treatment of mitomycin C and EVOOE reduced the drug cytotoxicity due to inhibition of ROS production. Conversely, the co‑treatment of T24 cells with paclitaxel and the polyphenol extract strongly increased the apoptotic cell death at each tested concentration compared to paclitaxel alone. Our results support the epidemiological evidence indicating that olive oil consumption exerts health benefits and may represent a starting point for the development of new anticancer strategies

Evaluation of the management of Hr-HPV+/PapTest- women. Results at 1-year recall

With cervical cancer screening the choice of 1-year as a period of follow-up in positive high-risk HPV women without cytological lesions is still under discussion. We evaluated the management of these women and the role of HPV genotyping test. We did a cervical cancer screening study of women aged 35-64 with primary high-risk HPV test. Women positive for high-risk HPV with negative cytology were followed-up after 1 year. In this study we selected women with high-risk HPV+/PapTest- resulted high-risk HPV+ at recall and performed the PapTest and HPV genotyping test. The detection rate of squamous high grade (CIN2+) relative to the total screened cohort was 2.1‰, and it was 0.2‰ at the 1-year recall. The colposcopy performed in women referred at the 1-year recall accounted for 48.8% of the total (baseline + 1-year recall), and 84.3% of these women had no cytological lesions. The most frequent hr-HPV genotype detected was HPV16 and 66.7% of co-infections were due to HPV16 and HPV18. 54.5% of women presented a persistent infection at 1-year recall with the same HPV subtype, 50% of persistent infections was due to HPV16 and 16.7% of these were determined to be CIN2+ histological lesions. Our data show that it may be useful to extend the period of follow-up for women hr-HPV+/PapTest- so as to reduce the number of unnecessary colposcopies due to the transitory infections and that the genotyping test could help to identify the persistent infections in which HPV16 is involved

Management of women aged 25-34 with diagnosis of ASCUS in the screening center of Latina.

In cervical cancer screening program of Latina (Italy) the hr-HPV as primary test is performed only on women aged 35-64 while women aged 25-34 are invited to perform PapTest. The aim of this study was to evaluate the impact of the application of the PapTest in women aged 25-34 and to evaluate the management of ASCUS. Women aged 25-34 were invited to perform PapTest according to the Italian guidelines; women with diagnosis of LSIL+ were referred to colposcopy while women with diagnosis of ASCUS were referred to hr-HPV test and only women resulted positive were referred to colposcopy. The 4.0% of women resulted positive to PapTest and the referral rate to colposcopy was 3.5%. The PPV value for CIN2+ at colposcopy was 7.2% and the Detection Rate (DR) for CIN2+ was 2.40‰. The ASCUS category was diagnosed in 41.8% of women resulted positive to PapTest and between them the 70.6% resulted positive to the hr-HPV test. The referral rate to colposcopy of women resulted positive to hr-HPV test was 1.1%. The PPV for CIN2+ at colposcopy and the DR of CIN2+ was 8.4% and 0.96‰ respectively. Between women with diagnosis of ASCUS, only 6 women showed a CIN2+ lesion (4 CIN2 and 2 CIN3). The present study showed that this algorithm, applied to women aged 25-34, obtained a good performance in term of test specificity (98%) and confirm that the application of hr-HPV test in the management of ASCUS leads to a decreased of inappropriate colposcopy due to transitory infection in young women

Near-Infrared Transflectance Spectroscopy Discriminates Solutions Containing Two Commercial Formulations of Botulinum Toxin Type A Diluted at Recommended Volumes for Clinical Reconstitution

: Botulinum neurotoxin type A (BoNT-A) is the active substance in pharmaceutical preparations widely used worldwide for the highly effective treatment of various disorders. Among the three commercial formulations of BoNT-A currently available in Italy for neurological indications, abobotulinum A toxin (Dysport\uae, Ipsen SpA, Milano, Italy) and incobotulinum A toxin (Xeomin\uae, Merz Pharma Italia srl, Milano, Italy) differ in the content of neurotoxin, non-toxic protein, and excipients. Clinical applications of BoNT-A adopt extremely diluted solutions (10-6 mg/mL) for injection in the target body district. Near-infrared spectroscopy (NIRS) and chemometrics allow rapid, non-invasive, and non-destructive methods for qualitative and quantitative analysis. No data are available to date on the chemometric analysis of the spectral fingerprints acquired from the diluted commercial formulations of BoNT-A. In this proof-of-concept study, we tested whether NIRS can categorize solutions of incobotulinum A toxin (lacking non-toxic proteins) and abobotulinum A toxin (containing non-toxic proteins). Distinct excipients in the two formulations were also analyzed. We acquired transmittance spectra in the visible and short-wave infrared regions (350-2500 nm) by an ASD FieldSpec 4\u2122 Standard-Res Spectrophotoradiometer, using a submerged dip probe designed to read spectra in transflectance mode from liquid samples. After preliminary spectra pre-processing, principal component analysis was applied to characterize the spectral features of the two BoNT-A solutions and those of the various excipients diluted according to clinical standards. Partial least squares-discriminant analysis was used to implement a classification model able to discriminate the BoNT-A solutions and excipients. NIRS distinguished solutions containing distinct BoNT-A commercial formulations (abobotulinum A toxin vs. incobotulinum A toxin) diluted at recommended volumes for clinical reconstitution, distinct proteins (HSA vs. incobotulinum A toxin), very diluted solutions of simple sugars (lactose vs. sucrose), and saline or water. Predictive models of botulinum toxin formulations were also performed with the highest precision and accuracy

A ribosomal protein is specifically recognized by saporin, a plant toxin which inhibits protein synthesis.

AbstractMany plants express enzymes which specifically remove an adenine residue from the skeleton of the 28 S RNA in the major subunit of the eukaryotic ribosome (ribosome inactivating proteins, RIPs). The site of action of RIPs (A4324 in the rRNA from rat liver) is in a loop structure whose nucleotide sequence all around the target adenine is also conserved in those species which are completely or partially insensitive to RIPs. In this paper we identify a covalent complex between saporin (the RIP extracted from Saponaria officinalis) and ribosomal proteins from yeast (Saccharomyces cerevisiae), by means of chemical crosslinking and immunological or avidin-biotin detection. The main complex (mol. wt. ≈ 60 kDa) is formed only with a protein from the 60 S subunit of yeast ribosomes, and is not detected with ribosomes from E. coli, a resistant species. This observation supports the hypotesis for a molecular recognition mechanism involving one or more ribosomal proteins, which could provide a ‘receptor’ site for the toxin and favour optimal binding of the target adenine A4324 to the active site of the RIP

ANALYSIS OF BIOCHEMICAL PROCESSES IN SINGLE LIVING CELLS BY QUANTITATIVE MICROSCOPY

Spectroscopic measurements in single living cells are made possible by the development of computer controlled light detectors, which, when applied to optical microscopes, yield spatial, temporal and eventually spectroscopic information about the sample. This minireview describes some experiments in which the distribution and concentration of specific intracellular markers (proteins, protein complexes, RNA) has been followed by quantitative microscopy. The examples chosen have contributed to shed light on a biochemical process as it happens in vivo; because of the non ideal conditions of the intracellular milieu, the comparison of in vivo and in vitro experiments is of great relevance to the understanding of cellular physiology

The crystal structure of saporin SO6 from Saponaria officinalis and its interaction with the ribosome

The 2.0 Angstrom resolution crystal structure of the ribosome inactivating protein saporin (isoform 6) from seeds of Saponaria officinalis is presented. The fold typical of other plant toxins is conserved, despite some differences in the loop regions. The loop between strands beta 7 and beta 8 in the C-terminal region which spans over the active site cleft appears shorter in saporin, suggesting an easier access to the substrate. Furthermore,ve investigated the molecular interaction between saporin and the yeast ribosome by differential chemical modifications. A contact surface inside the C-terminal region of saporin has been identified. Structural comparison between saporin and other ribosome inactivating proteins reveals that this region is conserved and represents a peculiar motif involved in ribosome recognition. (C) 2000 Federation of European Biochemical Societies