Observations on the water distribution and extractable sugar content in carrot slices after pulsed electric field treatment

peer-reviewedThe impact of pulsed electric field (PEF) processing conditions on the distribution of water in carrot tissue and extractability of soluble sugars from carrot slices was studied. Time domain NMR relaxometry was used to investigate the water proton mobility in PEF-treated carrot samples. Three distinct transverse relaxation peaks were observed in untreated carrots. After PEF treatment only two slightly-overlapping peaks were found; these were attributed to water present in the cytoplasm and vacuole of carrot xylem and phloem tissues. This post-treatment observation indicated an increase in water permeability of tissues and/or a loss of integrity in the tonoplast. In general, the stronger the electric field applied, the lower the area representing transverse relaxation (T2) values irrespective of treatment duration. Moreover an increase in sucrose, β- and α-glucose and fructose concentrations of carrot slice extracts after PEF treatment suggested increases in both cell wall and vacuole permeability as a result of exposure to pulsed electric fields.The authors acknowledge financial support from the Irish Phytochemical Food Network (IPFN) project funded under the Food Institutional Research Measure (FIRM, 06/TNI/AFRC6) of the Irish Department of Agriculture, Food and Marine. Dr. Aguiló-Aguayo thanks Generalitat of Catalonia for the postdoctoral grant Beatriu de Pinós (BP-DGR2010). E. Balagueró thanks the Lifelong Learning Programme for the internship grant Leonardo da Vinci MOTIVA3 (201 1-1-ES1-LEO02-34225)

Hypoxia-induced treatment failure in advanced squamous cell carcinoma of the uterine cervix is primarily due to hypoxia-induced radiation resistance rather than hypoxia-induced metastasis

Poor outcome of treatment in advanced cervix carcinoma has been shown to be associated with poor oxygenation of the primary tumour. Hypoxia may cause radiation resistance and promote lymph-node metastasis. The purpose of the study reported here was to investigate whether hypoxia-induced treatment failure in advanced cervix carcinoma is primarily a result of hypoxia-induced radiation resistance or the presence of hypoxia-induced lymph-node metastases at the start of treatment. Thirty-two patients with squamous cell carcinoma of the uterine cervix were included in the study. Radiation therapy was given with curative intent as combined external irradiation and endocavitary brachytherapy. The oxygenation status of the primary tumour was measured prior to treatment using the Eppendorf p O 2 Histograph. Pelvic and para-aortal lymph-node metastases were detected by magnetic resonance imaging at the time of initial diagnosis. The primary tumours of the patients with metastases (n = 18) were significantly more poorly oxygenated than those of the patients without metastases (n = 14). Multivariate Cox regression analyses involving biological and clinical parameters identified the tumour subvolume having p O 2 values below 5mmHg (HSV (p O 2< 5mmHg) as the only significant, independent prognostic factor for locoregional control, disease-free survival and overall survival. The probabillities of locoregional control, disease-free survival and overall survival were significantly lower for the patients with HSV (p O 2< 5 mmHg) above the median value than for those with HSV (p O 2< 5 mmHg) below the median value. On the other hand, the outcome of treatment was not significantly different for the patients with metastases and the patients without metastases at the start of treatment, irrespective of clinical end-point. Consequently, treatment failure was primarily a result of hypoxia-induced radiation resistance rather than hypoxia-induced lymph-node metastasis, suggesting that novel treatment strategies aiming at improving tumour oxygenation or enhancing the radiation sensitivity of hypoxic tumour cells may prove beneficial in attempts to improve the radiation therapy of advanced cervix carcinoma. © 2000 Cancer Research Campaig

Characterisation of invasive group B streptococci based on investigation of surface proteins and genes encoding surface proteins

ABSTRACTThe joint distributions of the six genes bca, bac, ɛ/alp1, alp2, alp3 and rib (encoding α-C-protein, β-C-protein, ɛ/Alp1, Alp2, Alp3, and Rib, respectively) and the proteins α-C-protein, β-C-protein and Rib were investigated in invasive isolates of group B streptococcus (GBS). In total, 297 invasive isolates (123 from neonates, 174 from adults) from south-west Sweden were collected during a 13-year period. Genes were detected using multiplex and specific PCRs, and expression of the surface proteins was demonstrated using monoclonal antibodies. The genes studied were found alone or in combinations in 294 (99%) of the invasive isolates. The most common genes were rib (n = 127 isolates, 43%), alp3 (n = 78, 26%) and ɛ/alp1 (n = 42, 14%). The bac gene was never found alone, but was found in combination with one other gene in 36 isolates. The surface proteins studied were detected alone or in combinations in 152 (51%) isolates, with the most common being Rib (n = 80, 27%), α-C-protein (n = 68, 23%) and β-C-protein (n = 24, 8%). Several genes were associated significantly with particular serotypes (e.g., ɛ/alp1 with serotype Ia; bca and bac with serotypes Ib and II; rib with serotype III; alp3 with serotype V). Overall, it was concluded that demonstration of different genes and surface proteins of GBS strains can be useful in epidemiological studies and in formulation of vaccines, but disappointingly, no single gene or surface protein included in the study was sufficiently common for it to be considered as the basis for a successful GBS vaccine

Raman Spectroscopic Mapping for the Analysis of Solar Radiation Induced Skin Damage

The effects of simulated solar irradiation of an artificial skin model have been examined using Raman spectroscopy and the results are compared with cytotoxicological and histological profiling. Samples exposed for times varying between 30 minutes and 240 minutes were incubated post exposure for a period of 96hours. The cytotoxicological response as measured by the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyl tetrazolium bromide] assay demonstrated a ~50% loss of viability of the artificial tissue after 120 minutes exposure. Histological staining of tissue sections showed considerable loss of cellular content in the epidermal layer at this endpoint. Raman spectroscopic mapping of tissue sections, coupled with K-means cluster analysis (KMCA) clearly identified the dermal and stratum corneum layers and differentiated further substructures of the epidermis. Post irradiation, a significant loss of DNA features in the basal layer was apparent in the results of the KMCA. Principal Components Analysis (PCA) of layers identified by the KMCA post exposure compared with controls indicated a significant increase in the lipidic signatures of the stratum corneum. In the dermal layer, little photodamage was observed, but a similar increase in lipidic signatures in the basal layer was accompanied by a decrease in DNA and protein contributions. The spectral profiles of the photodamage to the basal layer as identified by PCA are consistent over the exposure periods of 30-240 minutes, but an examination of the evolution of features associated with specific biochemical components indicated DNA damage and loss of lipidic signatures at the early exposure times, whereas changes in protein signatures appeared to evolve over longer periods. In comparison to the cytotoxicological responses, the study demonstrates that Raman spectroscopy can identify biochemical changes as a result of solar exposure at time points significantly earlier than changes in tissue viability are observed

Impact of pulsed electric field pre-treatment on nutritional and polyphenolic contents and bioactivities of light and dark brewer's spent grains

peer-reviewedPulsed electric field (PEF) pre-treatment, at 2.8 kV/cm with 3000 pulses of 20 μs pulse-width, was applied on brewer's spent grains (BSG) followed by aqueous extraction at 55 °C, 220 rpm for 16 h. PEF pre-treatment showed significantly increased yields (p 50 mg/mL) with lowest MIC value of 1.56 mg/mL against Staphylococcus aureus. All the BSG extracts induced the release of pro-inflammatory cytokines (IL-1β, IL-6, TNF-α) and chemokines (IL-8, MCP-1 and MIP-1α) confirming immunomodulatory activity.The authors acknowledge financial support from the ‘NovTechIng’ project funded under the Food Institutional Research Measure (Project No. FIRM/11/F/050) by the Irish Department of Agriculture, Food and Marine. The immunomodulatory study was performed using funding from the European Union's Horizon 2020 research and innovation programme under grant agreement No 634086 (NEPHSTROM). TPG is funded by a Hardiman Research Scholarship from the College of Medicine, Nursing and Health Sciences, National University of Ireland, Galway.The authors acknowledge financial support from the ‘NovTechIng’ project funded under the Food Institutional Research Measure (Project No. FIRM/11/F/050) by the Irish Department of Agriculture, Food and Marine. The immunomodulatory study was performed using funding from the European Union's Horizon 2020 research and innovation programme under grant agreement No 634086 (NEPHSTROM). TPG is funded by a Hardiman Research Scholarship from the College of Medicine, Nursing and Health Sciences, National University of Ireland, Galway

Genome-wide estimation of transcript concentrations from spotted cDNA microarray data

A method providing absolute transcript concentrations from spotted microarray intensity data is presented. Number of transcripts per µg total RNA, mRNA or per cell, are obtained for each gene, enabling comparisons of transcript levels within and between tissues. The method is based on Bayesian statistical modelling incorporating available information about the experiment from target preparation to image analysis, leading to realistically large confidence intervals for estimated concentrations. The method was validated in experiments using transcripts at known concentrations, showing accuracy and reproducibility of estimated concentrations, which were also in excellent agreement with results from quantitative real-time PCR. We determined the concentration for 10 157 genes in cervix cancers and a pool of cancer cell lines and found values in the range of 10(5)–10(10) transcripts per µg total RNA. The precision of our estimates was sufficiently high to detect significant concentration differences between two tumours and between different genes within the same tumour, comparisons that are not possible with standard intensity ratios. Our method can be used to explore the regulation of pathways and to develop individualized therapies, based on absolute transcript concentrations. It can be applied broadly, facilitating the construction of the transcriptome, continuously updating it by integrating future data