Gary Smerdon

The addition of hydrogen peroxide (H₂O₂) to cultured cells is widely used as a method to modulate redox-regulated cellular pathways, including the induction of programmed cell death in cell culture experiments and the testing of pro- and antioxidant compounds. Here, we assessed the effect on the cellular response to H₂O₂ of pre-adapting squamous cell carcinoma cells (A431) to the standard cell culture oxygenation of 18.6% O₂, compared to cells pre-adapted to a physiological skin O₂ concentration (3.0% O₂). We showed that cells pre-adapted to 18.6% O₂ resisted H₂O₂-induced cell death compared to cells pre-adapted to 3.0% O₂ for 96 h prior to treatment with H₂O₂. Moreover, the enzymatic activities of catalase and glutathione reductase, as well as the protein expression levels of catalase, were higher in cells pre-adapted to 18.6% O₂ compared to cells pre-adapted to 3.0% O_2. H₂O₂-resistant cells, pre-adapted to 18.6% O₂, exhibited increased nuclear Nrf-2 levels. It is concluded that A431 cells pre-adapted to standard cell culture oxygenation conditions resist H₂O₂-induced cell death. This effect may be related to their heightened activation of Nrf-2.

Access to deep-sea sponges brings with it the potential to discover novel antimicrobial candidates, as well as novel cold- and pressure-adapted bacteria with further potential clinical or industrial applications. In this study, we implemented a combination of different growth media, increased pressure and high-throughput techniques to optimize recovery of isolates from two deep-sea hexactinellid sponges, Pheronema carpenteri and Hertwigia sp., in the first culture-based microbial analysis of these two sponges. Using 16S rRNA gene sequencing for isolate identification, we found a similar number of cultivable taxa from each sponge species, as well as improved recovery of morphotypes from P. carpenteri at 22–25 °C compared to other temperatures, which allows a greater potential for screening for novel antimicrobial compounds. Bacteria recovered under conditions of increased pressure were from the phyla Proteobacteria , Actinobacteria and Firmicutes , except at 4 %O₂/5 bar, when the phylum Firmicutes was not observed. Cultured isolates from both sponge species displayed antimicrobial activity against Micrococcus luteus, Staphylococcus aureus and Escherichia coli .

Osteoradionecrosis of the mandible is the most common serious complication of radiotherapy for head and neck malignancy. For decades, hyperbaric oxygen has been employed in efforts to prevent those cases of osteoradionecrosis that are precipitated by dental extractions or implant placement. The evidence for using hyperbaric oxygen remains poor and current clinical practice varies greatly. We describe a protocol for a clinical trial to assess the benefit of hyperbaric oxygen in the prevention of osteoradionecrosis during surgery on the irradiated mandible. The HOPON trial is a phase III, randomised controlled, multi-centre trial. It employs an unblinded trial design, but the assessment of the primary endpoint, i.e. the diagnosis of osteoradionecrosis, is assessed on anonymised clinical photographs and radiographs by a blinded expert panel. Eligibility is through the need for a high-risk dental procedure in the mandible where at least 50-Gy radiotherapy has been received. Patients are randomised 1:1 to hyperbaric oxygen arm (Marx protocol) : control arm, but both groups receive antibiotics and chlorhexidine mouthwash. The primary endpoint is the presence of osteoradionecrosis at 6 months following surgery, but secondary endpoints include other time points, acute symptoms and pain, quality of life, and where implants are placed, their successful retention. The protocol presented has evolved through feasibility stages and through analysis of interim data. The classification of osteoradionecrosis has undergone technical refinement to ensure that robust definitions are employed. The HOPON trial is the only multi-centre RCT conducted in this clinical setting despite decades of use of hyperbaric oxygen for the prevention of osteoradionecrosis. European Clinical Trials Database, ID: EudraCT200700622527 . First registered on 5 November 2007.

The morphological similarity of Calanus species necessitates that the only unambiguous way of discriminating between the different species at any developmental stage is with molecular tools. We have developed a simple molecular technique to distinguish between the four species of Calanus copepods found in the North Atlantic (Calanus helgolandicus, C. finmarchicus, C. glacialis and C. hyperboreus) at any life stage. This system involves the PCR amplification of a region of the 16S rRNA gene, followed by Restriction Fragment Length Polymorphism (RFLP) analysis of the amplified product. This paper describes the application of the technique to a number of Calanus samples taken from a wide geographical range within the North Atlantic. Samples were acquired from sites ranging in latitude from the English Channel (50°N) to Tromsø (69°N), including many regions of co-occurrence. A comparison has been made between the distribution of Calanus as determined by molecular techniques and distributions established using traditional morphological identification. The molecular analysis has clearly shown extended areas of distribution and co-occurrence and has confirmed the intraspecific conservation of the restriction sites and therefore the reliability of the technique.