Developing a core outcome set for the health outcomes for children and adults with congenital oesophageal atresia and/or tracheo-oesophageal fistula: OCELOT task group study protocol.

Introduction: Heterogeneity in reported outcomes of infants with oesophageal atresia (OA) with or without tracheo-oesophageal fistula (TOF) prevents effective data pooling. Core outcome sets (COS) have been developed for many conditions to standardise outcome reporting, facilitate meta-analysis and improve the relevance of research for patients and families. Our aim is to develop an internationally-agreed, comprehensive COS for OA-TOF, relevant from birth through to transition and adulthood.

A novel anti-B7-H3 chimeric antigen receptor from a single-chain antibody library for immunotherapy of solid cancers.

B7-H3 (CD276) has emerged as a target for cancer immunotherapy by virtue of consistent expression in many malignancies, relative absence from healthy tissues, and an emerging role as a driver of tumor immune inhibition. Recent studies have reported B7-H3 to be a suitable target for chimeric antigen receptor-modified T cell (CAR-T) therapy using CARs constructed from established anti-B7-H3 antibodies converted into single-chain Fv format (scFv). We constructed and screened binders in an scFv library to generate a new anti-B7-H3 CAR-T with favorable properties.

Developing advanced nursing practice roles in cancer care.

Nurses in cancer care have been at the forefront of developing advanced nursing practice, however there has not been national agreement in relation to the education and clinical skills required for this practice. Work is under way by the NMC to develop a regulated framework for the standard for post-registration nursing.

Analysis of prostate tissue DNA for the presence of human papillomavirus by polymerase chain reaction, cloning, and automated sequencing.

We have analysed the DNA from 24 prostate tissue biopsies, spanning a range of Gleason grading from benign to grade 5 and mixed randomly with cervical cancer samples of known human papillomavirus (HPV) status, for the prevalence of HPV DNA, in a double-blind study to ensure complete objectivity. Polymerase chain reactions (PCR) were performed using general E1 open reading frame primers for HPV under low stringency conditions, in addition to reactions containing primers specific for HPV16, E2, and E6 open reading frames under higher, more stringent PCR conditions. The presence of cellular DNA was verified by the use of primers for hypoxanthine guanine phosphoribosyl transferase.

Functional expression of sv40 in normal human prostatic epithelial and fibroblastic cells - differentiation pattern of nontumorigenic cell-lines.

To study mesenchymal-epithelial interactions associated with the normal and pathological human prostate, we have developed a model of well differentiated human prostate epithelial and fibroblastic cells. Normal human prostatic cells, either of epithelial or fibroblastic origins were successfully transfected with SV40 and strains with extended lifespan were selected until the crisis was reached, within 20 and 30 passages for the epithelial and fibroblastic cells, respectively. Only a few clones emerged from the crisis: PNT1A (Cussenot et al: J Urol 143: 881-886, 1991), PNT1B and PNT2 epithelial cell lines.

Is the mammalian cell plasma membrane a barrier to oxygen transport?

Oxygen transport in the Chinese hamster ovary (CHO) plasma membrane has been studied by observing the collision of molecular oxygen with nitroxide radical spin labels placed in the lipid bilayer portion of the membrane at various distances from the membrane surface using the long-pulse saturation-recovery electron spin resonance (ESR) technique. The collision rate was estimated for 5-, 12-, and 16-doxylstearic acids from spin-lattice relaxation times (T1) measured in the presence and absence of molecular oxygen. Profiles of the local oxygen transport parameters across the membrane were obtained showing that the oxygen diffusion-concentration product is lower than in water for all locations at 37 degrees C.

Radiation induction of drug resistance in RIF-1: correlation of tumor and cell culture results.

The RIF-1 tumor line contains cells that are resistant to various anti-neoplastic drugs, including 5-fluorouracil (5FU), methotrexate (MTX), adriamycin (ADR), and etoposide (VP16). The frequency of these drug-resistant cells is increased after irradiation. The frequency of drug-resistant cells and the magnitude of radiation-induced drug resistance are different in cell culture than in tumors.

DNA content in nasopharyngeal carcinoma.

DNA analysis by flow cytometry was performed on tissue blocks from 41 patients with nasopharyngeal carcinoma. The histologic slides were reviewed by a pathologist and blindly classified according to the World Health Organization classification. The paraffin-embedded blocks were processed to obtain individual nuclei, which were then stained with propidium iodide.

Flow cytometric evaluation of chemosensitive and chemoresistant head and neck tumors.

For patients with head and neck squamous carcinoma, a clinical response to induction chemotherapy has correlated with a survival advantage. Similarly, patients with diploid tumors have displayed a survival advantage when compared with patients with aneuploid tumors. This study examined DNA content in 33 patients who had undergone induction chemotherapy as part of two clinical protocols to determine if there was a correlation between the patients with diploid tumors and the patients with a clinical response to chemotherapy.

Drug resistance following irradiation of RIF-1 tumors: influence of the interval between irradiation and drug treatment.

RIF-1 tumors contain a small number of cells (1 to 100 per 10(6) cells) that are resistant to 5-fluorouracil, methotrexate, or adriamycin. The frequency of drug-resistant cells among individual untreated tumors is highly variable. Radiation, delivered in vivo at doses of 3 to 12 Gy, increases the frequency of methotrexate- and 5-fluorouracil-resistant cells, but not the frequency of adriamycin-resistant cells.

Radiation induction of drug resistance in RIF-1 tumors and tumor cells.

The RIF-1 tumor cell line contains a small number of cells (1-20 per 10(6) cells) that are resistant to various single antineoplastic drugs, including 5-fluorouracil (5FU), methotrexate (MTX), and adriamycin (ADR). For 5FU the frequency of drug resistance is lower for tumor-derived cells than for cells from cell culture; for MTX the reverse is true, and for ADR there is no difference. In vitro irradiation at 5 Gy significantly increased the frequency of drug-resistant cells for 5FU, MTX, and ADR.

Cytofluorometric analysis of the DNA content in ovarian carcinoma and its relationship to patient survival.

Forty-one patients with epithelial malignancies of the ovary treated at the Medical College of Wisconsin Affiliated Hospitals from 1976 to 1984 had paraffin embedded tissue available for review. Of the 41 patients, 40 had adequate material to provide 50 micron sections that were evaluated with flow cytometry to determine DNA content. Tumor- and patient-related parameters were then correlated with the results of flow cytometry.

Mechanisms of membrane damage for CHO cells heated in suspension.

Chinese hamster ovary cells were heated for 20 min at 45.5 degrees C in different conditions, and quantitative determinations of cellular membrane blebbing were performed for cells maintained at 25 degrees C and 37 degrees C after hyperthermia. The percentage of cells with blebs following heating was dependent upon the composition of the medium during heating and the posthyperthermia temperature after heating.

An evaluation of paramagnetic broadening agents for spin probe studies of intact mammalian cells.

Six transition metal ion complexes have been examined for their effects on the cell survival as well as their effectiveness in inducing the broadening of the electron spin resonance (ESR) spectra of nitroxide spin probes. These paramagnetic species are Ni(EDTA), Ni(DTPA), potassium tris(oxalato) chromate (chromium oxalate), K3Fe(CN)6, Cu(DTPA), and NiCl2. At 100 mM concentration, the typical concentration used in cell studies to broaden the extracellular nitroxide ESR signal, only Ni(EDTA) and Ni(DTPA) are found to be non-toxic to Chinese hamster ovary cells.

Cytotoxicity of commonly used nitroxide radical spin probes.

Since nitroxide radical spin probes are used frequently to test biophysical properties of cells, their use should be restricted to conditions that do not perturb normal cell growth and viability. Eight commonly used nitroxide radical spin probes have been tested for their effects on the survival of CHO cells. These include water-soluble spin probes Tempol, Tempamine, CTPO, CTPC and 4-maleimido-Tempo, and lipid soluble spin probes 5-Doxyl-, 12-Doxyl-, and 16-Doxylstearates.

Changes in bleb formation following hyperthermia treatment of Chinese hamster ovary cells.

Chinese hamster ovary cells in suspension cultures were heated for various times at 41.5, 43.5, and 45.

Effect of melanin on radiation response of CHO cells.

The effect of the presence of melanin on the response of mammalian cells to ionizing radiation was investigated in a model system utilizing the ability of Chinese hamster ovary cells to incorporate melanin by endocytosis. Cells were incubated in monolayer cultures from 2 to 20 hours with melanin prepared from 'beef eye' or synthesized by air oxidation of 3,4-dihydroxyphenylalanine. For asynchronous cultures, the survival curve parameters for cells incubated with both types of melanin were indistinguishable from those of the same cells without added melanin.

Effect of hypoxia on recovery from damage induced by heat and radiation in plateau-phase CHO cells.

The effect of hypoxia on the induction of and recovery from damage by radiation alone and in combination with heat has been investigated using plateau-phase Chinese hamster ovary (CHO) cells. Postirradiation hypoxia reduced the potentially lethal damage recovery (PLDR) in cells irradiated under an euoxic state and completely eliminated PLDR in cells irradiated under hypoxia. Cells which were maintained under hypoxia during both irradiation and a 4-hr recovery period and then incubated for a further period of 4 hr under euoxic conditions showed PLDR, suggesting that the inhibition of PLDR by hypoxia is reversible.

Membrane fluidity of Chinese hamster ovary cells on plasma fibronectin-coated microcarriers. A spin label study.

Purified plasma fibronectin promotes the spreading of Chinese hamster ovary (CHO) cells on microcarriers in a serum-free medium. The promotion of 50% of cell spreading on microcarriers requires about 3.4 X 10(8) fibronectin molecules per bead.

Melanin endocytosis by cultured mammalian cells. A model for melanin in a cellular environment.

The incorporation of natural eumelanin from bovine eyes and synthetic 3,4-dihydroxy-phenylalanine (dopa) melanin into Chinese hamster ovary (CHO) cells is reported. The process is linear for at least 8 h. Electron microscopy showed phagocytosis of melanin, either as a single granule or in groups of granules, into cell lysosomes with subsequent degradation of the granule.

Inhibition of repair of premutational lesions in plateau phase Chinese hamster cells exposed to gamma radiation.

Induction and repair of forward mutations to 8-azaguanine resistance were studied in gamma irradiated, plateau phase Chinese hamster ovary cells. Mutation induction increased with dose with a relatively low induction for doses below 4 Gy and a steep increase thereafter. A close correlation between the ability of radiation to induce both lethality and mutations in plateau phase cells was evident.

Modification of mutation frequency in plateau phase Chinese hamster ovary cells exposed to gamma radiation during recovery from potentially lethal damage.

Induction of forward mutations leading to 8-azaguanine resistance was studied in plateau phase Chinese hamster ovary cells exposed to gamma-radiation. Survival and mutation frequency were assessed immediately after irradiation or after 6 hours of post-irradiation recovery at 37 degrees C. Cells exposed to doses above 4Gy showed appreciable increase in the survival level as a result of recovery from potentially lethal damage.