Coulomb excitation of 104Sn and the strength of the 100Sn shell closure.

A measurement of the reduced transition probability for the excitation of the ground state to the first 2+ state in 104Sn has been performed using relativistic Coulomb excitation at GSI. 104Sn is the lightest isotope in the Sn chain for which this quantity has been measured. The result is a key point in the discussion of the evolution of nuclear structure in the proximity of the doubly magic nucleus 100Sn. The value B(E2; 0+ → 2+) = 0.10(4) e2b2 is significantly lower than earlier results for 106Sn and heavier isotopes. The result is well reproduced by shell model predictions and therefore indicates a robust N = Z = 50 shell closure.

Expression of genes involved in early cell fate decisions in human embryos and their regulation by growth factors.

Little is understood about the regulation of gene expression in human preimplantation embryos. We set out to examine the expression in human preimplantation embryos of a number of genes known to be critical for early development of the murine embryo. The expression profile of these genes was analysed throughout preimplantation development and in response to growth factor (GF) stimulation. Developmental expression of a number of genes was similar to that seen in murine embryos (OCT3B/4, CDX2, NANOG). However, GATA6 is expressed throughout preimplantation development in the human. Embryos were cultured in IGF-I, leukaemia inhibitory factor (LIF) or heparin-binding EGF-like growth factor (HBEGF), all of which are known to stimulate the development of human embryos. Our data show that culture in HBEGF and LIF appears to facilitate human embryo expression of a number of genes: ERBB4 (LIF) and LIFR and DSC2 (HBEGF) while in the presence of HBEGF no blastocysts expressed EOMES and when cultured with LIF only two out of nine blastocysts expressed TBN. These data improve our knowledge of the similarities between human and murine embryos and the influence of GFs on human embryo gene expression. Results from this study will improve the understanding of cell fate decisions in early human embryos, which has important implications for both IVF treatment and the derivation of human embryonic stem cells.

Binding of sodium dodecyl sulfate and hexaethylene glycol mono-n-dodecyl ether to the block copolymer L64: electromotive force, microcalorimetry, surface tension, and small angle neutron scattering investigations of mixed micelles and polymer/micellar surfactant complexes.

The interactions of sodium dodecyl sulfate (SDS) with the triblock copolymer L64 (EO13-PO30-EO13) and hexaethylene glycol mono-n-dodecyl ether (C12EO6) were studied using electromotive force, isothermal titration microcalorimetry, differential scanning microcalorimetry, and surface tension measurements. In certain regions of binding, mixed micelles are formed, and here we could evaluate an interaction parameter using regular solution theory. The mixed micelles of L64 with both SDS and C12EO6 exhibit synergy. When L64 is present in its nonassociated state, it forms polymer/micellar SDS complexes at SDS concentrations above the critical aggregation concentration (cac). The cac is well below the critical micellar concentration (cmc) of pure SDS, and a model suggesting how bound micelles are formed at the cac in the presence of a polymer is described. The interaction of nonassociated L64 with C12EO6 is a very rare example of strong binding between a nonionic surfactant and a nonionic polymer, and C12EO6/L64 mixed micelles are formed. We also carried out small angle neutron scattering measurement to determine the structure of the monomeric polymer/micellar SDS complex, as well as the mixed L64/C12EO6 aggregates. In these experiments, contrast matching was achieved by using the h and d forms of SDS, as well as C12EO6. During the early stages of the formation of polymer-bound SDS micelles, SDS aggregates with aggregation numbers of approximately 20 were found and such complexes contain 4-6 bound L64 monomers. The L64/C12EO6 data confirmed the existence of mixed micelles, and structural information involving the composition of the mixed micelle and the aggregation numbers were evaluated.

The union doctor.

The appointment, terms of service, and work of the Poor Law medical officer, the union doctor, is described to illustrate one of the roles of the mid-nineteenth century general practitioner. It was this role which laid the foundation of modern general practice.

Richard Quiller Couch--an outstanding nineteenth century general practitioner.

The boom in metal mining in the mid-nineteenth century produced a fearful mortality among miners. The history of this problem is described by the life and work of Mr R. Quiller Couch, a Cornish mine surgeon and general practitioner. He was a remarkable man and an outstanding general practitioner.

The rise of the general practitioner in the nineteenth century.

It was the Apothecaries Act of 1815 which led to the emergence of general practice as we know it today and it was this one Act which produced a flood of changes on the medical scene that are without parallel in our history. Students were soon to undergo new forms of training in new medical schools and hospitals, and many medical associations and journals were founded. The term ;general practitioner' was soon in use. The driving force behind all these changes was the Society of Apothecaries and the new general practitioners, and all too often they were opposed by the two Royal Colleges. It was only at the beginning of the twentieth century that these new practitioners were allowed to call themselves ;doctors'.

The certifying surgeons.

The origin and work of the nineteenth century certifying surgeons is described. Their place within the broad spectrum of general practice is emphasized and their efforts to improve the welfare of children is noted.

Club practice.

There were many forms of club or contract practice in the nineteenth century, but the friendly societies were the most important. A brief history of the friendly societies is given. As they grew in numbers and importance so did the dissatisfaction of the doctors who worked with them. Discontent among the doctors led at the end of the century to a battle between the medical profession and the clubs. The issues which divided the clubs and the doctors were clearly defined but, although the battle was protracted, the doctors did not win or manage to change the system of medical provision for the poor. The club system was ended by Lloyd George when he introduced his National Insurance Act, 1911.

Very high level overexpression of bacterial elongation factor Tu.

The tufA gene, which encodes the bacterial elongation factor Tu of E. coli, was isolated as a 1226 bp DNA fragment using the polymerase chain reaction(1). EcoR1 restriction enzyme digestion sites were incorporated at the 3' and 5' termini of the gene. The tufA gene was ligated into the plasmid vector pUC19 and was sequenced fully. A plasmid shown to contain the tufA gene in the correct orientation with respect to the lac promoter of pUC19 was then used to transform E.coli strain JM105 and expression of EFTu in induced cultures of the resulting clone, JM105 pJBDB01, was measured using a continuous monitoring procedure. The clone was shown to produce up to 95 mg of bacterial elongation factor Tu per litre of cell culture.

Expression of cell adhesion molecules during human preimplantation embryo development.

Formation of a fully differentiated, implantation competent blastocyst requires the expression of a complex repertoire of molecules. However, the events that drive morphogenesis are poorly elucidated in the human embryo. In this work, we describe the amplification of representative cDNAs from morphologically and developmentally normal, individual human embryos at all stages from pronucleate to blastocyst. These cDNAs were probed to reveal the temporal expression pattern of cell adhesion molecules thought to play a key role in murine preimplantation embryo development. We demonstrated constitutive expression of beta actin, beta 1 and alpha 6 integrins, ZO-1 and E-cadherin, as shown previously in mouse embryos. No expression of beta 3, alpha 2, alpha 3 or alpha 7 integrins nor of L or P selectin was detected at any stage of preimplantation development. beta 5 integrin showed a regulated pattern of expression and was not expressed in blastocysts, while desmocollin-2 could only be detected at the blastocyst stage. Expression and localization of beta 1, beta 5 and alpha 6 integrins and ZO-1 and E-cadherin proteins was confirmed in blastocyst stage embryos by immunocytochemistry. We have identified differences in the expression of integrin molecules between mouse and human embryos, and propose a role for alpha v beta 5 and alpha 6 beta 1 integrin dimers in the human embryo at implantation.

Improved methods of cultivation and production of deuteriated proteins from E. coli strains grown on fully deuteriated minimal medium.

AIMS: The aim was to develop reliable and economical protocols for the production of fully deuteriated biomolecules by bacteria. This required the preparation of deuterium-tolerant bacterial strains and an understanding of the physiological mechanisms of acquisition of deuterium tolerance. METHODS AND RESULTS: We report here improved methods for the cultivation of Escherichia coli on fully deuteriated minimal medium. A multi-stage adaptation protocol was developed; this included repeated plating and selection of colonies and resulted in highly deuterium-tolerant cell cultures. Three E. coli strains, JM109, MRE600 and MRE600Rif, were adapted to growth on deuteriated succinate medium. This is the first report of JM109 being adapted to deuteriated minimal media. The adapted strains showed good, consistent growth rates and were capable of being transformed with plasmids. Expression of heterologous proteins in these strains was reliable and yields were consistently high (100-200 mg l-1). We also show that all E. coli cells are inherently capable of growth on deuteriated media. CONCLUSIONS: We have developed a new adaptation protocol that resulted in three highly deuterium-tolerant E. coli strains. Deuterium-adapted cultures produced good yields of a deuteriated recombinant protein. We suggest that E. coli cells are inherently capable of growth on deuteriated media, but that non-specific mutations enhance deuterium tolerance. Thus plating and selection of colonies leads to highly deuterium-tolerant strains. SIGNIFICANCE AND IMPACT OF STUDY: An understanding of the mechanism of adaptation of E. coli to growth on deuteriated media allows strategies for the development of disabled deuterium-tolerant strains suitable for high-level production of deuteriated recombinant proteins and other biomolecules. This is of particular importance for nuclear magnetic resonance and neutron scattering studies of biomolecules.

Binding of sodium dodecyl sulfate to linear and star homopolymers of the nonionic poly(methoxyhexa(ethylene glycol) methacrylate) and the polycation poly(2-(dimethylamino)ethyl methacrylate): electromotive force, isothermal titration calorimetry, surface tension, and small-angle neutron scattering measurements.

We investigated the binding of sodium dodecyl sulfate (SDS) to various linear and star polymers of the nonionic methoxyhexa(ethylene glycol) methacrylate (PMHEGMA) and the ionic 2-(dimethylamino)ethyl methacrylate (PDMAEMA), the latter being a polycation at low pH. The dodecyl sulfate ion selective electrode (EMF), isothermal titration calorimetry (ITC), and surface tension (ST) were applied to gain detailed information about interactions. In all cases there is evidence of significant binding of SDS over an extensive SDS concentration range spanning from ca. 10(-6) to 0.1 mol dm(-3). At pH 3, the polymer PDMAEMA is a strong polycation and here the binding is dominated by electrostatic 1:1 charge neutralization with the anionic surfactant. At their natural pH of 8.6, PMHEGMA and PDMAEMA polymers are essentially nonionic and bind SDS in the form of polymer-bound aggregates in the concentration range of ca. 1 x 10(-3) to 3 x 10(-2) mol dm(-3). All the polymers also bind SDS to a lesser extent at concentrations below 1 x 10(-3) mol dm(-3) reaching as low as 10(-7) mol dm(-3). This low concentration binding process involves the polymer and nonassociated SDS monomers. As far as we are aware, this is the first example that such a low concentration noncooperative binding process could be observed in SDS/neutral polymer systems by EMF and ST. We also showed that the nonionic surfactant hexa(ethylene glycol) mono-n-dodecyl ether (C12EO6) and the cationic cetyltrimethylammonium bromide (C16TAB) interact with star PDMAEMA. We believe that the interaction of C12EO6 and CTAB is of similar noncooperative type as the first SDS binding process in the range from ca. 10(-5) to 0.3 x 10(-3) mol dm(-3). At the high concentration binding limit Csat of SDS, the above polymers become fully saturated with bound SDS micelles. We applied small angle neutron scattering (SANS) to determine the structure and aggregation numbers of the star polymer/bound SDS micelles and calculated the stoichiometry of such supramolecular complexes. The SANS data on PDMAEMA star polymers in the presence of C12EO6 showed only a limited monomer binding in contrast to linear PDMAEMA, which showed monomer C12EO6 binding at low concentrations but micellar aggregates at 6 x 10(-3) mol dm(-3).