Highly efficient and sensitive patient-specific quality assurance for spot-scanned proton therapy.

The purpose of this work was to develop an end-to-end patient-specific quality assurance (QA) technique for spot-scanned proton therapy that is more sensitive and efficient than traditional approaches. The patient-specific methodology relies on independently verifying the accuracy of the delivered proton fluence and the dose calculation in the heterogeneous patient volume. A Monte Carlo dose calculation engine, which was developed in-house, recalculates a planned dose distribution on the patient CT data set to verify the dose distribution represented by the treatment planning system. The plan is then delivered in a pre-treatment setting and logs of spot position and dose monitors, which are integrated into the treatment nozzle, are recorded. A computational routine compares the delivery log to the DICOM spot map used by the Monte Carlo calculation to ensure that the delivered parameters at the machine match the calculated plan. Measurements of dose planes using independent detector arrays, which historically are the standard approach to patient-specific QA, are not performed for every patient. The nozzle-integrated detectors are rigorously validated using independent detectors in regular QA intervals. The measured data are compared to the expected delivery patterns. The dose monitor reading deviations are reported in a histogram, while the spot position discrepancies are plotted vs. spot number to facilitate independent analysis of both random and systematic deviations. Action thresholds are linked to accuracy of the commissioned delivery system. Even when plan delivery is acceptable, the Monte Carlo second check system has identified dose calculation issues which would not have been illuminated using traditional, phantom-based measurement techniques. The efficiency and sensitivity of our patient-specific QA program has been improved by implementing a procedure which independently verifies patient dose calculation accuracy and plan delivery fidelity. Such an approach to QA requires holistic integration and maintenance of patient-specific and patient-independent QA.

Evaluation of a dual-arm Archimedean spiral array for microwave hyperthermia.

PURPOSE: This effort describes a third-party performance evaluation of a novel, commercial, dual-armed Archimedean spiral array hyperthermia applicator. The applicator is analysed for its ability to couple efficiently into muscle equivalent phantom loads, operate over a broad bandwidth to help accommodate variable tissue properties and generate predictable and repeatable SAR contours that are adaptable to clinically probable disease shapes. MATERIALS AND METHODS: Characterization of the applicator includes E-field and return-loss measurements in liquid muscle tissue-equivalent phantom, as well as comparison of 'treatment-planning' simulations of several possible array SAR patterns with measured SAR from non-coherently driven spiral array antennae. RESULTS: The applicator demonstrates a reasonably low return loss over a large bandwidth and the ability to generate a very uniform heating pattern. Ability to adjust SAR contours spatially to fit specific shapes is also demonstrated. CONCLUSIONS: This device should prove a welcome addition to a currently limited set of superficial heating applicators to provide controllable heating of superficial tissue disease.

Graded and areal expression patterns of regulatory genes and cadherins in embryonic neocortex independent of thalamocortical input.

The differentiation of areas of the mammalian neocortex has been hypothesized to be controlled by intrinsic genetic programs and extrinsic influences such as those mediated by thalamocortical afferents (TCAs). To address the interplay between these intrinsic and extrinsic mechanisms in the process of arealization, we have analyzed the requirement of TCAs in establishing or maintaining graded or areal patterns of gene expression in the developing mouse neocortex. We describe the differential expression of Lhx2, SCIP, and Emx1, representatives of three different classes of transcription factors, and the type II classical cadherins Cad6, Cad8, and Cad11, which are expressed in graded or areal patterns, as well as layer-specific patterns, in the cortical plate. The differential expression of Lhx2, SCIP, Emx1, and Cad8 in the cortical plate is not evident until after TCAs reach the cortex, whereas Cad6 and Cad11 show subtle graded patterns of expression before the arrival of TCAs, which later become stronger. We find that these genes exhibit normal-appearing graded or areal expression patterns in Mash-1 mutant mice that fail to develop a TCA projection. These findings show that TCAs are not required for the establishment or maintenance of the graded and areal expression patterns of these genes and strongly suggest that their regulation is intrinsic to the developing neocortex.

Defects in thalamocortical axon pathfinding correlate with altered cell domains in Mash-1-deficient mice.

We have analyzed the pathfinding of thalamocortical axons (TCAs) from dorsal thalamus to neocortex in relation to specific cell domains in the forebrain of wild-type and Mash-1-deficient mice. In wild-type mice, we identified four cell domains that constitute the proximal part of the TCA pathway. These domains are distinguished by patterns of gene expression and by the presence of neurons retrogradely labeled from dorsal thalamus. Since the cells that form these domains are generated in forebrain proliferative zones that express high levels of Mash-1, we studied Mash-1 mutant mice to assess the potential roles of these domains in TCA pathfinding. In null mutants, each of the domains is altered: the two Pax-6 domains, one in ventral thalamus and one in hypothalamus, are expanded in size; a complementary RPTP(delta) domain in ventral thalamus is correspondingly reduced and the normally graded expression of RPTP(delta) in that domain is no longer apparent. In ventral telencephalon, a domain characterized in the wild type by Netrin-1 and Nkx-2.1 expression and by retrogradely labeled neurons is absent in the mutant. Defects in TCA pathfinding are localized to the borders of each of these altered domains. Many TCAs fail to enter the expanded, ventral thalamic Pax-6 domain that constitutes the most proximal part of the TCA pathway, and form a dense whorl at the border between dorsal and ventral thalamus. A proportion of TCAs do extend further distally into ventral thalamus, but many of these stall at an aberrant, abrupt border of high RPTP(delta) expression. A small proportion of TCAs extend around the RPTP(delta) domain and reach the ventral thalamic-hypothalamic border, but few of these axons turn at that border to extend into the ventral telencephalon. These findings demonstrate that Mash-1 is required for the normal development of cell domains that in turn are required for normal TCA pathfinding. In addition, these findings support the hypothesis that ventral telencephalic neurons and their axons guide TCAs through ventral thalamus and into ventral telencephalon.

Older rural women and the use of complementary therapies.

One hundred seventy-five rural women over the age of 70 participated in a study to assess their use of complementary therapies and practitioners. Data were collected via questionnaires and interviews. Findings suggest that these women used nontraditional health practices to treat a variety of symptoms, as well as to maintain their independence. However, very few of them informed their primary care providers that they used complementary therapies and practitioners. The majority were inadequately informed regarding the efficacy and safety of the therapies.

Partially folded states of the capsid protein of cowpea severe mosaic virus in the disassembly pathway.

The different partially folded states of the capsid protein that appear in the disassembly pathway of cowpea severe mosaic virus (CPSMV) were investigated by examining the effects of hydrostatic pressure, sub-zero temperatures and urea. The conformational states of the coat protein were analyzed by their intrinsic fluorescence, binding of bis(8-anilinonaphthalene-1-sulfonate) (bis-ANS) and susceptibility to trypsin digestion. CPSMV could be disassembled by pressure at 2.5 kbar. Intrinsic fluorescence and hydrodynamic measurements showed that pressure-induced dissociation was completely reversible. Virus pressurization in the presence of ribonuclease revealed that viral RNA was not exposed, since it was not digested by the enzyme, suggesting the maintenance of protein-nucleic acid interactions under pressure. When the temperature was decreased to -10 degrees C under pressure, CPSMV disassembly became an irreversible process and in this condition, viral RNA was completely digested by ribonuclease. These results suggest a relationship between protein-RNA interactions and CPSMV assembly. Bis-ANS binding and trypsin digestion of coat proteins revealed that they assume a different conformation when they are denatured by low temperatures under pressure or than when they are denatured by urea at atmospheric pressure. The results indicate that the coat proteins can exist in at least four states: (1) The native conformation in the virus capsid; (2) bound to RNA when the virus is dissociated by pressure at room temperature, assuming a conformation that retains the information for reassembly; (3) free subunits in a molten-globule conformation when the virus is dissociated by low temperature under pressure; and (4) free subunits completely unfolded by high concentrations of urea.

Induction and repression of mammalian achaete-scute homologue (MASH) gene expression during neuronal differentiation of P19 embryonal carcinoma cells.

MASH1 and MASH2, mammalian homologues of the Drosophila neural determination genes achaete-scute, are members of the basic helix-loop-helix (bHLH) family of transcription factors. We show here that murine P19 embryonal carcinoma cells can be used as a model system to study the regulation and function of these genes. MASH1 and MASH2 display complementary patterns of expression during the retinoic-acid-induced neuronal differentiation of P19 cells. MASH1 mRNA is undetectable in undifferentiated P19 cells but is induced to high levels by retinoic acid coincident with neuronal differentiation. In contrast, MASH2 mRNA is expressed in undifferentiated P19 cells and is repressed by retinoic acid treatment. These complementary expression patterns suggest distinct functions for MASH1 and MASH2 in development, despite their sequence homology. In retinoic-acid-treated P19 cells, MASH1 protein expression precedes and then overlaps expression of neuronal markers. However, MASH1 is expressed by a smaller proportion of cells than expresses such markers. MASH1 immunoreactivity is not detected in differentiated cells displaying a neuronal morphology, suggesting that its expression is transient. These features of MASH1 expression are similar to those observed in vivo, and suggest that P19 cells represent a good model system in which to study the regulation of this gene. Forced expression of MASH1 was achieved in undifferentiated P19 cells by transfection of a cDNA expression construct. The transfected cells expressing exogenous MASH1 protein contained E-box-binding activity that could be super-shifted by an anti-MASH1 antibody, but exhibited no detectable phenotypic changes. Thus, unlike myogenic bHLH genes, such as MyoD, which are sufficient to induce muscle differentiation, expression of MASH1 appears insufficient to promote neurogenesis.

Progressive relaxation and the sleep of older noninstitutionalized women.

Fifty-five randomly selected women over the age of 65 years participated in this study to determine the effect of progressive relaxation on their sleep patterns. Using a Pretest-Posttest Same Subject design, subjects spent 8 nights participating in the study. Progressive relaxation was introduced on days 4 and 5. Data were collected with polysomnography and the Sleep Pattern Questionnaire. It was found that progressive relaxation significantly improved certain sleep parameters. Nurses should encourage its use with older women who report disturbed sleep patterns and sleep dissatisfaction.

Hemorrhagic colitis and pseudomelanosis coli in ipecac ingestion by proxy.

This report describes a toddler with chronic diarrhea, vomiting, and hypotonia due to surreptitious administration of syrup of ipecac by his mother (Munchausen's syndrome by proxy). Several features of this case distinguish it from previous reports of chronic ipecac ingestion in childhood: the development of grossly bloody stools; radiologic, endoscopic, and biopsy evidence of a chronic moderate colitis resembling ulcerative colitis; and the histologic finding of pseudomelanosis coli, providing an important clue to toxic ingestion. The significance and possible mechanism for genesis of pseudomelanosis coli is discussed. This case emphasizes the variability in presentation and difficulty in diagnosing long-term ipecac ingestion by proxy. Ipecac toxicity should be considered in children with unexplained colitis and vomiting.

Structural studies of bean pod mottle virus, capsid, and RNA in crystal and solution states by laser Raman spectroscopy.

Structures of protein and RNA components of bean pod mottle virus (BPMV) have been investigated by use of laser Raman spectroscopy. Raman spectra were collected from both aqueous solutions and single crystals of BPMV capsids (top component) and virions (middle and bottom components, which package, respectively, small and large RNA molecules). Analysis of the data permits the assignment of conformation-sensitive Raman bands to viral protein and RNA constituents and observation of structural similarities and differences between solution and crystalline states of BPMV components. The Raman results show that the protein subunits of the empty capsid contain between 45% and 55% beta-strand and beta-turn secondary structure, in agreement with the recently determined X-ray crystal structure, and that this total beta-strand content undergoes a small increase (approximately 5%) with packaging of RNA. The subunits are relatively deficient in alpha-helix secondary structure, estimated at less than 25%, and therefore must contain extensive amounts (greater than 20%) of loops and irregular chain conformations. The Raman spectra also show the following: (1) The molecular environments of as many as four tryptophan residues per subunit are altered upon packaging RNA, resulting in stronger 1N-H hydrogen bonding for two Trp residues and more hydrophobic environments for two indole rings. (2) Hydrogen-bonding states of the seven Tyr residues per subunit do not change detectably when RNA is packaged. At least five tyrosine OH groups are involved exclusively as strong hydrogen bond donors to protein acceptor groups, which suggests restricted access of solvent H2O molecules to these parahydroxyls.(ABSTRACT TRUNCATED AT 250 WORDS)

Comparison of two automated instrument systems for rapid susceptibility testing of gram-negative bacilli.

The Vitek AutoMicrobic System with GSC-plus cards and the Abbott MS-2 system were tested in parallel and the results were compared directly with those of a reference microdilution minimal inhibitory concentration (MIC) procedure on a group of 262 clinical isolates of the family Enterobacteriaceae and of Pseudomonas aeruginosa. Results of both systems were compared with the reference MIC for category agreement, and in addition, the Vitek MICs were compared with those obtained by the reference procedure. The Vitek system provided an essential category correlation of 89.4% for enteric bacteria and 97.0% for P. aeruginosa. Vitek MICs agreed within 1 twofold dilutional increment for 86.3% of the enteric bacteria tested and for 96.2% of the P. aeruginosa isolates. The Abbott MS-2 essential categoric agreement was 92.0% for enteric bacteria and 92.4% for P. aeruginosa. If only aminoglycosides or carbenicillin were considered for P. aeruginosa isolates, the essential category agreement was 92.5% for the Vitek and 93.3% for the MS-2. The majority of MS-2 category errors (13 of 19) with P. aeruginosa involved gentamicin results on isolates whose reference MICs were 8 micrograms/ml and whose MS-2 results were susceptible (MIC less than or equal to 4 micrograms/ml). Retesting of the P. aeruginosa isolates in calcium-supplemented MS-2 broth increased the essential agreement for the aminoglycosides to 97.5%.

The effects of family constellation and child gender on parental use of evaluative feedback.

Mothers and fathers of 43 middle-class families were observed individually interacting during a paper-folding task with either their only or middle child in order to assess parental use of evaluation and task-facilitative behaviors with preschoolers. Middle children were from 3-child families in which the older sibling was the same sex as the target child. Sibling spacing was greater than 36 months in 12 families and less than 36 months in 11 families. With total interaction, parental income, education, and age as control variables, analyses indicated that parental approvals, disapprovals, task facilitative feedback, and helping behaviors varied with parent gender, child gender, and family constellation. Children's task performance did not differ significantly among groups. It was concluded that gender effects on parental use of evaluation feedback in parent-child teaching interactions may be different for families with different types of family structure.

The effects of cognitive and behavioral control on coping with an aversive health examination.

The effects of cognitive and behavioral control on coping with an aversive health examination were tested in a 2 x 2 factorial design. Cognitive control was represented by sensory or health-education information; instruction or no instruction in abdominal relaxation constituted the levels of behavioral control. The study was conducted in the natural setting of a family-planning clinic with 24 young women who were undergoing a routine pelvic examination. Subjects who received sensory information prior to the examination showed less distress, as indicated by overt distress behaviors and pulse rates, than did subjects who received health-education information. Cognitive control did not show a significant effect on self-report of fear. No significant effects were demonstrated for the factor of behavioral control. The results suggested that cognitive control information, which emphasizes the sensory experiences typically accompanying an aversive event, limits reactivity to aversive stimuli. The reduction in reactivity is thought to result in an increased ability to cope with aversive events.

A fine structural study of degenerative changes in the dorsal column nuclei of aging mice. Lack of protection by vitamin E.

The dorsal column nuclei of young, old, and vitamin E-supplemented old mice were examined by light and electron microscopy. Evidence of neuroaxonal dystrophy (NAD) was found in young (3-mo.-old) mice and increased with age. Vitamin E, added to the diet in the amount of 0.3%, did not protect the nuclei from age-associated degeneration. The NAD was characterized by enlarged profiles containing patches of smooth reticular networks and groups of vesicles. Various stages of mitochondrial alteration, producing multivesicular bodies as intermediate stages, were found, and other unusual forms of dense bodies were also observed. Axons, synaptic terminals, and possibly glial cells were affected, and, by 23 mo. of age, a large number of nerve fibers in nucleus gracilis were dystrophic, while nucleus cuneatus was affected to a lesser extent.