Unsupervised explorative data analysis of normal human leukocytes and BCR/ABL positive leukemic cells mid-infrared spectra.

We proved the ability of Fourier Transform Infrared microspectroscopy (microFTIR) complemented by Principal Component Analysis (PCA) to detect protein phosphorylation/de-phosphorylation in mammalian cells. We analyzed by microFTIR human polymorphonuclear neutrophil (PMNs) leukocytes, mouse-derived parental Ba/F3 cells (Ba/F3#PAR), Ba/F3 cells transfected with p210(BCR/ABL) (Ba/F3#WT) and expressing high levels of protein tyrosine kinase (PTK), and human-derived BCR/ABL positive K562 leukemic cell sub-clones engineered to differently express receptor-type tyrosine-protein phosphatase gamma (PTPRG). Synchrotron radiation (SR) and conventional (globar) IR sources were used to perform microFTIR respectively, on single cells and over several cells within the same sample. Ex vivo time-course experiments were run, inducing maximal protein phosphorylation in PMNs by 100 nM N-formylated tripeptide fMLP. Within the specific IR fingerprint 1800-850 cm(-1) frequency domain, PCA identified two regions with maximal signal variance. These were used to model and test the robustness of PCA in representing the dynamics of protein phosphorylation/de-phosphorylation processes. An IR signal ratio marker reflecting the homeostatic control by protein kinases and phosphatases was identified in normal leukocytes. The models identified by microFTIR and PCA in normal leukocytes also distinguished BCR/ABL positive Ba/F3#WT from BCR/ABL negative Ba/F3#PAR cells as well as K562 cells exposed to functionally active protein tyrosine phosphatase recombinant protein ICD-Tat transduced in cells by HIV-1 Tat technology or cells treated with the PTK inhibitor imatinib mesylate (IMA) from cells exposed to phosphatase inactive (D1028A)ICD-Tat recombinant protein and untreated control cells, respectively. The IR signal marker correctly reflected the degrees of protein phosphorylation associated with abnormal PTK activity in BCR/ABL positive leukemic cells and in general was inversely related to the expression/activity of PTPRG in leukemic sub-clones. In conclusion, we have described a new, reliable and simple spectroscopic method to study the ex vivo protein phosphorylation/de-phosphorylation balance in cell models: it is suitable for biomedical and pharmacological research labs but it also needs further optimization and its evaluation on large cohorts of patients to be proposed in the clinical setting of leukemia.

Complex systems, part I: why 42 is rarely, if ever, the ultimate answer.

We describe the different categories of systems and systems thinking, and illustrate why almost all clinical interactions constitute simultaneously complex and complicated systems, so-called "wicked systems". We also discuss why they are not amenable to quantitative analysis. With the use of comparisons and illustrations we show some of the problems that reductionist metrics create, and support concerns regarding quantitative fallacy. The systematic correlation of data in medicine was one of the earliest achievements of the discipline. Recently, however, the overwhelming bias towards an evidence base, which relies almost entirely on randomised controlled trials, has created a reductionist view that often excludes important aspects of medicine and, in particular, surgery. We must now move away from thinking that is controlled by the "tyranny of metrics" to embrace complex-systems thinking, and work across disciplines. We outline the arguments for this and give clinical examples from oral and maxillofacial surgery.

Expression analysis of TFIID in single human oocytes: new potential molecular markers of oocyte quality.

Molecular characterization of human female gametes should make it easier to understand the basis of certain infertility disorders. Biologically significant mRNAs have been analysed in single oocytes to search for molecular biomarkers of oocyte quality. Initial analysis was focused on mRNA for proteins involved in cell growth and cycle control, specifically those encoding members of the general transcription apparatus such as the subunits of the general transcription factor TFIID. This heteromultimeric protein, comprising about 15 subunits, is the most important general transcription factor of the second class. These proteins are essential for the initiation of transcription of protein-coding genes, so they must be present in mature oocytes for mRNA synthesis during the first phases of embryonic development. Semi-quantitative reverse transcriptionpolymerase chain reaction was used to identify different TFIID subunits in single oocytes and to search for differences in expression as compared with control tissues. The data show that the mRNAs for most TFIID subunits are indeed synthesized in oocytes, but their expression profiles differ markedly. TATAbox-binding protein associated factor 4B (TAF4B), TAF5 and TATAbox-binding protein-like 2 (TBPL2) are expressed at higher levels in oocytes than in control tissues. It is suggested that they could be used as biomarkers of oocyte quality.

Making and working of a new electronic resource for patients, carers and professionals: maxfacts.uk.

Many maxillofacial patients have serious short, medium, or long-term problems, as well as having to make informed decisions about often life-changing interventions. Validated comprehensive information, at the right time and the right level for a diverse group of users (patients, carers, and professionals), is vital if patients are to make a serious contribution to their treatment. We describe the development of an online resource for this purpose. Maxfacts.uk aims to cover every aspect of oral and maxillofacial surgery and care, from neck dissection and ballistic wounds to physiotherapy and texture-modified foods. The principles of design, and the multilayered structure, interface, and functions of such a multiuser website are outlined, including accessibility and engagement. The maxfacts model and design could easily be adapted for other users with similar, complex, long-term needs.

Ternary graph as a questionnaire: a new approach to assessment of quality of life?

This study was prompted by awareness of the importance of research into quality of life (QoL) for patients with diseases of the head and neck, the important part questionnaires currently play in this field, and awareness of the "questionnaire fatigue" experienced by many patients. Our multidisciplinary research group raised coincidental awareness of the widespread use of ternary graphs in the sciences, social sciences, and humanities as a graphical tool for quantitative, semiquantitative, or purely graphical characteristics of ternary mixtures. We explored how the basic properties of ternary graphs could be translated into an interactive electronic tool as an alternative to conventional questionnaires. We have described how this was done, and offered open access to an interactive ternary-graph based (self) assessment tool, specifically designed for the needs of patients with conditions of the head and neck. Finally, have we made open-source code available for those who may wish to adapt or develop the tool for further applications.