Energetics of Storage and Diffusion of Water and Cyclo-Octasulfur for a Nonpolar Cavity of RHCC Tetrabrachion by Molecular Dynamics Simulations.

Tetrabrachion forms the key component of the S-layer of Staphylothermus marinus. Molecular dynamics simulations have been used to study the energetics of occupancy of cavity 3 of the right-handed coiled-coil stalk of tetrabrachion by both water molecules and cyclooctasulfur S 8 crowns, as well as to determine possible pathways and free energy barriers for the diffusion of both water and cyclooctasulfur through the peptide walls of RHCC tetrabrachion between cavity 3 and bulk solvent. Calculations of the transfer free energy from solvent to cavity show that clusters of six, seven and eight water molecules are marginally stable in cavity 3, but that occupancy of the cavity by a cyclooctasulfur ring is favoured significantly over water clusters of all sizes. Thermal activation simulations at T = 400K revealed that water molecules diffusing through the wall pass through a sequence of metastable configurations where they are temporarily immobilized by forming networks of hydrogen bonds with specific wall residues. Calculations of the free energy of these metastable configurations using multi-configurational thermodynamic integration yielded a free energy profile with a principal free energy maximum ∆G~50 kJ/mol and a slight activation asymmetry in favour of the direction from cavity to solvent. Potential exit pathways for cyclooctasulfur were investigated with the methods of steered molecular dynamics and umbrella sampling. The cyclooctasulfur was steered through a gap in the tetrabrachion wall along a linear path from cavity 3 into the solvent and the resulting trajectory was subdivided into 22 sampling windows. The free energy profile created for the trajectory by umbrella sampling showed a sharp principal maximum as a function of the reaction coordinate with asymmetric free energy barriers ∆G exit ~220 kJ/mol and ∆G entrance ~100 kJ/mol for cavity exit and entrance, respectively.

A 64-channel transmitter for investigating parallel transmit MRI.

Multiple channel radiofrequency (RF) transmitters are being used in magnetic resonance imaging to investigate a number of active research topics, including transmit SENSE and B(1) shimming. Presently, the cost and availability of multiple channel transmitters restricts their use to relatively few sites. This paper describes the development and testing of a relatively inexpensive transmit system that can be easily duplicated by users with a reasonable level of RF hardware design experience. The system described here consists of 64 channels, each with 100 W peak output level. The hardware is modular at the level of four channels, easily accommodating larger or smaller channel counts. Unique aspects of the system include the use of vector modulators to replace more complex IQ direct digital modulators, 100 W MOSFET RF amplifiers with partial microstrip matching networks, and the use of digital potentiometers to replace more complex and costly digital-to-analog converters to control the amplitude and phase of each channel. Although mainly designed for B(1) shimming, the system is capable of dynamic modulation necessary for transmit SENSE by replacing the digital potentiometers controlling the vector modulators with commercially available analog output boards. The system design is discussed in detail and bench and imaging data are shown, demonstrating the ability to perform phase and amplitude control for B(1) shimming as well as dynamic modulation for transmitting complex RF pulses.

Overcoming phase effects of voxel-sized coils in planar and cylindrical arrays.

Parallel imaging in MRI (using multiple coils to partially encode k-space) is currently the primary route to decreasing scan time. Single echo acquisition (SEA) imaging is a completely parallel imaging method recently developed by our group that collects a full image in a single echo. Phase encoding is eliminated and replaced by the spatial localization of long and very narrow coils. The fact that the coils are on the order of the voxel size for the first time in an MR application has led to an examination of the effect of the phase of the coil on the signal received from the voxel and how to most effectively manipulate it. Obtaining full signal from a voxel in planar arrays can be accomplished with a single gradient compensation pulse, but the phase effects in cylindrical arrays are more complex due to the changing coil angle with regard to the Cartesian axes of the gradients. This paper discusses the signal-phase interactions of planar and cylindrical arrays of voxel-sized coils and suggests methods for phase manipulation for optimization.

Analogs of guanine nucleoside triphosphates for sequencing applications.

We have synthesized more than 30 different deoxyribonucleosides and triphosphates with modifications either in the base or the phosphate moiety as analogs of 2'-dGTP for DNA sequencing applications. All the modified nucleoside triphosphates were tested as substrates for DNA polymerases, including Sequenase T7 DNA polymerase or Thermo Sequenase DNA polymerase. Two of the analogs, 7-ethyl-7-deaza-dGTP and 7-hydroxymethyl-7-deaza-dGTP meet our requirements as better sequencing reagents.

Conducting qualitative research in the health sector: researcher issues and dilemmas.

This article contributes to bridging the gap between research activity and the practical implementation of management decision making in the health sector by reflecting upon some of the issues and dilemmas for researchers, whether academics or managers, in conducting qualitative research in this sector. The article presents the methodological issues addressed by a team of researchers engaged on a project about manager learning and development in an NHS Trust, and highlights concerns about ethical issues that emerged from the research process. The study had involved a series of interviews with senior managers and clinical staff, doctors, nurses and therapists, and it addressed issues surrounding change within the organization, the impact on individuals' jobs, and the resultant learning and development required and undertaken. The article emphasizes that engaging in reflection on the research process is valuable and suggests that it should become a mainstream part of such research. It concludes that there is an important role of qualitative management research in the health sector and that for it to be acceptable and valued, it must be operationally sensitive, ethically robust and methodologically rigorous.

Consensus radiation protection practices for academic research institutions.

Under the auspices of the Howard Hughes Medical Institute, a set of consensus guidelines for Radiation Protection Practices has been developed for biomedical research using radioactive materials. The purposes of the guidelines are (1) to promote good radiation protection practices consistent with the needs of biomedical research, the ALARA principle, and regulatory requirements; (2) to establish common goals and consistent practices within radiation safety programs; and (3) to build a meaningful partnership between radiation safety professionals and the biomedical research community. These practices are intended to enhance radiation protection and the efficiency of the research staff. The consensus guidelines will lessen the variability in radiation safety practices that is evident among many academic research institutions and will encourage better acceptance and regulatory compliance by users of radioactive materials in biomedical research.

Howard Hughes Medical Institute dose assessment survey.

Biomedical science researchers often express frustration that health physics practices vary widely between individual institutions. A survey examining both internal and external dose assessment practices was devised and mailed to fifty institutions supporting biomedical science research. The results indicate that health physics dose assessment practices and policies are highly variable. Factors which may contribute to the degree of variation are discussed.

Substituted O6-benzylguanine derivatives and their inactivation of human O6-alkylguanine-DNA alkyltransferase.

Several new O6-benzylguanine analogs bearing increasingly bulky substituent groups on the benzene ring or at position 9 were tested for their ability to inactivate the human DNA repair protein, O6-alkylguanine-DNA alkyltransferase. Substitution on the benzene ring was well tolerated although activity varied considerably with structural changes in groups attached to position 9. For this site, activity was preserved with large or small lipophilic groups while introduction of non-carbohydrate polar groups generally reduced activity regardless of their size.

Structural features of substituted purine derivatives compatible with depletion of human O6-alkylguanine-DNA alkyltransferase.

A series of O6- and S6-substituted purine derivatives were tested for their ability to deplete the human DNA repair protein O6-alkylguanine-DNA alkyltransferase (AGT) in cell-free extracts from HT29 colon tumor cells and intact HT29 cells. The order of potency was O6-(p-Y-benzyl)-guanine (Y = H, F, Cl, and CH3) > O6-benzyl-2'-deoxyguanosine > O6-(p-Y-benzyl)guanosine (Y = H, Cl, and CH3) > or = a series of 9-substituted O6-benzylguanine derivatives > or = O6-allylguanine > O6-benzylhypoxanthine > O6-methylguanine. A series of 7-substituted O6-benzylguanine derivatives, 2-amino-6-(p-Y-benzylthio)purine (Y = H, CH3), 2-amino-6-[(p-nitrobenzyl)thio]-9-beta-D-ribofuranosylpurine, and 7-benzylguanine were inactive. It is concluded that for efficient AGT depletion, an allyl or benzyl group attached through exocyclic oxygen at position 6 of a 2-aminopurine derivative is required. Activity is preserved with a variety of substituent groups attached to position 9 while substitution at position 7 leads to a complete loss of activity.

Ultrastructural morphometric study of follicular center lymphocytes: II. Analyses of cleaved-cell populations do not support the Lukes-Collins' concept.

Ultrastructural morphometric analysis was carried out on six cases of lymph node biopsies with reactive hyperplasia to establish the frequency and depth of invaginations in nuclear profiles situated in the mantle zones and follicular centers. The frequency distribution of the depth of invaginations was similar in nuclear profiles whether in the small lymphocytes of mantle zones or the small, partially transformed (centrocytes) and fully transformed (centroblasts) lymphocytes of follicular centers. Invaginated and cleaved lymphocytes were not confined to the partially transformed (centrocytic) lymphocytes of follicular centers, and nuclear profiles with invaginations bore no resemblance to those depicted in the Lukes-Collins model. A considerable proportion of mantle zone lymphocyte nuclear profiles had invaginations (ranging from 7.5% to 53.6%) and there was no difference between the frequency of deep indentations or clefts in mantle zone lymphocytes (8.1 +/- 5.4%) and the small unstimulated (9.3 +/- 5.3%) and partially transformed (8.4 +/- 1.4%) lymphocytes in follicular centers. Computer modeling of stylized nuclei with conical indentations indicated that all lymphocytic nuclei likely have multiple invaginations or groove-like creases.

Thiopental bolus during carotid endarterectomy-rational drug therapy?

Ten studies were performed to examine the time course of arterial and venous thiopental concentrations following the administration of thiopental (4 mg X kg-1 over 3 min) for cerebral protection during carotid occlusion in nine patients undergoing elective carotid endarterectomy; in five patients the time course of EEG change was also studied. The arterial and venous thiopental concentrations were similar with no evidence of a sustained arterial-venous gradient. The average arterial concentration was 20.1 microgram X ml-1 +/- 10 (SD) at 2 min after thiopental, and fell rapidly to 13.0 micrograms X ml-1 +/- 3.2 at 5 min, 10.7 micrograms X ml-1 +/- 4.4 at 10 min and 6.2 micrograms X ml-1 at 30 min. After thiopental the EEG record showed an increase in delta activity and in four patients a burst suppression pattern was seen. The duration of burst suppression activity was variable (130 to 367 seconds) but in all instances cortical activity had returned to the pre-thiopental level by five to ten minutes. Thus concentrations of thiopental of 10-30 micrograms X ml-1 were associated with EEG burst suppression and both were seen only within the first five minutes after drug administration. In contrast the carotid artery was occluded for considerably longer (26 +/- 4) minutes. We conclude that, since there was no sustained arterial-venous gradient, either arterial or venous concentrations are adequate for the study of thiopental pharmacokinetics.(ABSTRACT TRUNCATED AT 250 WORDS)

The value of smears in the diagnosis of gonorrhoea in the female.

Of 332 consecutive patients attending the female venereal disease clinic, Auckland Hospital, 74 patients had gonorrhoea confirmed by culture, but gram stained slides were positive in only 36 of these patients. Five other patients had positive gram stained smears but were culture negative. The study showed the value of gram stained smear in the rapid diagnosis of gonorrhoea in the female.

Histologic changes in the seminiferous tubules after vasectomy.

PIP: 4 different studies on the effect of vasectomy on the testis in canines and humans are reported. Testicular biopsies were performed at certain intervals and the results in both humans and dogs were nearly identical. It was found that spermatogenesis 2 to 3 weeks after vasectomy remained unchanged with accumulation of spermatozoa in the tubules. Between 3 and 6 weeks, progressive spermatogenic arrest with few spermatozoa and decreased spermatids were observed. Between 100 and 300 days, occasional mature sperm were found in the tubules indicating a return of spermatogenesis. Meiotic studies showed this to be an arrest in early prophase. It is theorized that spermatogenesis may be sensitive to pressure changes in the tubular system.^ieng