bluuebunny/crossref_metadata_2025 huggingface dataset:
A randomized controlled trial of botulinum toxin on lower limb spasticity following acute acquired severe brain injury
Authors: Verplancke D , Snape S , Salisbury C F , Jones P W , Ward A B
Published: February 2005

Objective: To determine whether serial casting combined with botulinum toxin reduces the development of calf contracture after severe head injury.

Design: A double-blind placebo-controlled trial of three parallel treatments for lower limb spasticity.

Setting: Acute general hospital in the UK.

Subjects: Adults aged 17–70 years admitted to hospital following a severe brain injury.

Interventions: Current physical treatment (group I), lower leg casting plus injections with either saline (group II), or with botulinum toxin (group III) into gastrocnemius and soleus muscles.

Measures: Limit of ankle dorsiflexion at entry and exit after up to 12 weeks, the Glasgow Outcome Scale (GOS) and Modified Ashworth Scale (MAS).

Results: Two hundred and fifty-three patients were screened and 35 were entered into the trial. Three patients died and four failed to complete the trial. Eighty-eight per cent of those entering the randomized part of the study developed spasticity within 14 days of their injury and the mean range of improvement in the angle of passive ankle dorsiflexion was 4.598 in controls, 11.698 in cast and saline and 13.598 in cast and botulinum toxin. There were significant improvements in the MAS scores in actively treated groups, but not in controls. Cast and botulinum toxin patients also demonstrated a significant improvement in the GOS.

Conclusions: Active intervention with casting prevents talipes equinovarus deformities in patients losing ankle movement following severe brain injury. Casting alone in these patients is sufficient; the role of additional botulinum toxin needs further investigation, but is safe in these patients.
Is it possible to actively and purposely make use of plasticity and adaptability in the neurorehabilitation treatment of multiple sclerosis patients? A pilot project
Authors: Rasova K , Krasensky J , Havrdova E , Obenberger J , Seidel Z , Dolezal O , Rexova P , Zalisova M
Published: February 2005

Objective: To investigate whether neurorehabilitation is able to influence clinical parameters and brain function measured radiologically.

Design: A group of healthy probands was compared with two groups of multiple sclerosis (MS) patients, one of which received rehabilitative therapy.

Setting: Outpatient in a university hospital.

Subjects: Twenty-eight patients with multiple sclerosis (MS), 17 of whom received rehabilitative therapy, and 13 healthy controls.

Interventions: Two months of rehabilitative eclectic therapy based on principles of sensorimotor learning and adaptation.

Main measures: Multiple Sclerosis Functional Composite, Modified Fatigue Impact Scale, Beck Depression Inventory Score, Barthel Index, Environment Status Scale and Multiple Sclerosis Quality of Life-54, and functional magnetic resonance imaging (fMRI).

Results: Patients who underwent neurorehabilitation showed a greater drop in fatigue, depression, impairment, disability and handicap and more improvement in quality of life than those who did not receive therapy. Correlation of brain activity between the right and the left hemisphere is greater in healthy individuals than in MS patients. Neurorehabilitation resulted in a trend for increased correlation between the left and the right hemisphere in patients (approaching the standard). In comparison with control groups, signal amplitudes in anatomical areas did not show any significant changes.

Conclusion: Clinical changes seen with neurorehabilitation were not associated with any detectable changes in fMRI observations.
Improving patient and carer communication, multidisciplinary team working and goal-setting in stroke rehabilitation
Authors: Monaghan J , Channell K , McDowell D , Sharma A K
Published: February 2005

Objective: To determine the extent to which three forms of multidisciplinary team (MDT) care in stroke rehabilitation meet the standards set by the United Kingdom National Service Framework (NSF).

Design: Consecutive assessment of the three forms of care was completed.

Subjects: The study included three groups of 25 stroke inpatients on the stroke rehabilitation ward.

Intervention: (1) A standard weekly MDT meeting using a standard form for documentation; (2) a standard MDT meeting using a newly devised form; and (3) a novel MDT ward round using the new form, and attended by doctors.

Results: MDT ward rounds result in significantly better consideration of patients' needs (median 7 per patient compared with 0 and 5 in phases one and two), enhanced SMART (specific, measurable, achievable, realistic and time framed) goal-setting (median 3 per patient compared to 1 in phases one and two); greater patient involvement (12 patients compared to 0 and 4 in phases one and two); and improved team working (measured using the team climate inventory) than do MDT meetings.

Conclusions: In the present study, standard weekly MDT meetings did not meet the standards set for MDT care by the NSF. The use of a MDT ward round allows these standards to be achieved.
Sustained attention in patients with mild traumatic brain injury
Authors: Chan R CK
Published: February 2005

Objective: To demonstrate that two tests of sustained attention were sensitive to attention deficits in patients with mild traumatic brain injury (TBI).

Design: A cross-sectional study recruiting 51 patients with TBI and 51 matched controls.

Outcome measures: The Sustained Attention to Response Task (SART) and Monotone Counting Test.

Results: The patient groups performed significantly worse than the normal controls in both sustained attention tests. The SART performance was also correlated with pathology severity in the patient group in terms of loss of consciousness ( r=0.247, p=0.05). A cut-off of less than 1 standard deviation (SD) gives optimal diagnostic information in terms of sensitivity in the present sample (0.61 for Monotone Counting Test; 0.75 for SART).

Conclusion: These findings suggest that the SART and Monotone Counting Test are sensitive to patients with mild TBI. The SART-assessed sustained attention is also sensitive enough to detect attention impairment in this clinical group regardless of diagnosis and may provide clinicians with an alternative method of assessing sustained attention in these clinical groups.
A pilot study of the effect of aerobic exercise on people with moderate disability multiple sclerosis
Authors: Kileff J , Ashburn A
Published: February 2005

Objective: A pilot study to investigate the effect of aerobic exercise on the mobility and function of people with moderate disability multiple sclerosis (MS).

Design: A small group, comparative, pre-and post-intervention study.

Setting: A gymnasium within a general hospital.

Subjects: Eight subjects, all female, average age 45 years (range 33-61) with moderate disability MS (Kurtzke scale 4-6), were recruited; six completed the study.

Intervention: Intervention consisted of bi-weekly sessions of 30 min cycling on a static bike at their maximal level of exertion for 12 weeks.

Main measures: The stability of each individual's condition was established pre-intervention using three baseline assessments over two weeks. A battery of tests (the 10-metre and 6-min walk tests, the Functional Reach, the Gulick and the Guys Neurological Disability Scale) were completed pre and post intervention. Potential negative effects were recorded on the Fatigue Severity Scale and the Modified Ashworth Scale. An independent rater completed assessments post intervention. Inter-rater reliability was found to be acceptable.

Results: Comparisons pre and post intervention (Wilcoxon signed ranked test) showed significant improvement on the Guys Neurological Disability Scale (p = 0.026), with the mean score reducing from 13 to 9, and the 6-min walk test (p = 0.046), with the mean distance increasing from 200 m to 261 m. Other measures failed to reach significance.

Conclusions: Findings from this pilot study suggest overall disability and mobility improved with the aerobic training. A positive treatment effect in this small study suggests the need for a larger trial. Knowledge of how to establish predictive heart rate and how to monitor the effects of cycling with people with MS will be used to inform future clinical trials.
A study to assess the effect of nursing interventions at the weekend for people with stroke
Authors: Davidson Ian , Hillier Valerie F , Waters Karen , Walton Timothy , Booth Joanne
Published: February 2005

Objective: To examine whether additional therapy provided by nurses at the weekend improved the physical outcome for people with stroke on a stroke rehabilitation unit.

Design: A single blind randomized controlled trial.

Setting: A 16-bed stroke rehabilitation unit in the north of England.

Subjects: Forty-one people with stroke were randomized by means of minimization to intervention and control groups.

Interventions: The intervention group received additional exercise at the weekend provided by the nursing staff and the control group received their usual care. Both groups received usual care during weekdays.

Main outcome measures: The Motor Assessment Scale (MAS), the Barthel Index (BI) and length of stay in hospital.

Results: No significant differences were found between the groups in terms of MAS and BI at discharge but there was a borderline significant difference between the groups on unconditional testing in terms of length of stay in hospital and on the stroke unit ( p = 0.05 and p = 0.07 respectively). However, these findings were in favour of the control group. On conditional testing (adjusting for BI on admission and age) these differences disappeared ( p = 0.14 and p = 0.15) for length of stay in hospital and on the stroke unit respectively.

Conclusions: The present study indicates that an increase in one-to-one input by nurses for people with stroke did not lead to a measurable difference in outcome in this small study.
Vocational perspectives after spinal cord injury
Authors: Schönherr M C , Groothoff J W , Mulder G A , Eisma W H
Published: February 2005

Objective: To give insight into the vocational situation several years after a traumatic spinal cord injury (SCI) and describe the personal experiences and unmet needs; to give an overview of health and functional status per type of SCI and their relationship with employment status.

Design: Descriptive analysis of data from a questionnaire.

Setting: Dutch rehabilitation centre with special department for patients with spinal cord injuries.

Subjects: Fifty-seven patients with a traumatic SCI, aged 18-60 years, admitted to the rehabilitation centre from 1990 to 1998.

Main measures: Questionnaire with items related to vocational outcome, job experiences, health and functional status.

Results: Of 49 patients who were working at the moment of SCI 60% currently had a paid job. Vocational outcome was related to a higher educational level. A significant relation between the SCI-specific health and functional status and employment was not found. The respondents who changed to a new employer needed more time to resume work, but seemed more satisfied with the job and lost fewer working hours than those who resumed work with the same employer. In spite of reasonable to good satisfaction with the current work situation, several negative experiences and unmet needs were reported.

Conclusions: Despite a high participation in paid work following SCI, the effort of the disabled worker to have and keep a job should not be underestimated.
Efficacy of Bobath versus orthopaedic approach on impairment and function at different motor recovery stages after stroke: a randomized controlled study
Authors: Wang Ray-Yau , Chen Hsiu-I , Chen Chen-Yin , Yang Yea-Ru
Published: February 2005

Objective: To investigate the effectiveness of Bobath on stroke patients at different motor stages by comparing their treatment with orthopaedic treatment.

Design: A single-blind study, with random assignment to Bobath or orthopaedic group.

Setting: Physical therapy department of a medical centre.

Subjects: Twenty-one patients with stroke with spasticity and 23 patients with stroke at relative recovery stages participated.

Interventions: Twenty sessions of Bobath programme or orthopaedic treatment programme given in four weeks.

Main outcome measures: Stroke Impairment Assessment Set (SIAS), Motor Assessment Scale (MAS), Berg Balance Scale (BBS) and Stroke Impact Scale (SIS) for impairment and functional limitation level.

Results: Participants with spasticity showed greater improvement in tone control (change score: 1.209±1.03 versus 0.089±0.67, p =0.006), MAS (change score: 7.64±4.03 versus 4.009±1.95, p =0.011), and SIS (change score: 7.309±6.24 versus 1.259±5.33, p=0.023) after 20 sessions of Bobath treatment than with orthopaedic treatment. Participants with relative recovery receiving Bobath treatment showed greater improvement in MAS (change score: 6.149±5.55 versus 2.779±9.89, p=0.007), BBS (change score: 19.189±15.94 versus 6.859±5.23, p=0.015), and SIS scores (change score: 8.509±3.41 versus 3.629±4.07, p =0.006) than those with orthopaedic treatment.

Conclusion: Bobath or orthopaedic treatment paired with spontaneous recovery resulted in improvements in impairment and functional levels for patient with stroke. Patients benefit more from the Bobath treatment in MAS and SIS scores than from the orthopaedic treatment programme regardless of their motor recovery stages.
Comparison of the impact of a single session of isokinetic or isotonic muscle stretch on gait in patients with spastic hemiparesis
Authors: Maynard V , Bakheit A MO , Shaw S
Published: February 2005

Objective: To establish if isokinetic and isotonic muscle stretch (with or without weight-bearing) of the ankle plantar flexors improves gait in hemiplegic patients. A further aim was to compare the effectiveness of these treatment methods.

Design: A randomized, parallel group prospective study.

Setting: A stroke rehabilitation unit.

Subjects: Ambulatory hemiparetic stroke patients with mild to moderately severe muscle hypertonia of the lower limb and a group of healthy control subjects.

Intervention: Subjects were randomized to receive a single 20-min session of isokinetic muscle stretch or isotonic muscle stretch with or without weight-bearing.

Outcome measures: Selected kinematic, kinetic and spatio-temporal gait parameters were measured at baseline, immediately after the muscle stretch and 24 h later.

Results: Sixty-six patients and 21 healthy control subjects were recruited and completed the study. There were statistically significant differences between the patient groups and the healthy subjects on most of the gait parameters studied. However, the differences between the patient groups or between the three measurements over time for each type of muscle stretch did not reach statistical significance.

Conclusions: A single session of isokinetic or isotonic muscle stretch (with or without weight-bearing) of the ankle plantar flexors has no clinically observable effect on the gait of hemiplegic stroke patients.
Effect of recommended positioning on stroke outcome at six months: a randomized controlled trial
Authors: Jones A , Tilling K , Wilson-Barnett J , Newham D J , Wolfe C DA
Published: February 2005

Objective: To evaluate the effect on patient outcome of a teaching package for nurses designed to improve the positioning of stroke patients.

Design: Cluster randomized controlled trial with six-month follow-up.

Setting: Ten stroke rehabilitation hospital units located within one UK inner city region. These were randomized to control or intervention group.

Subjects: A sample of 120 patients admitted within four weeks of a first stroke and with a hemiplegia. No eligible patient refused to participate. Eighty-three (69%) completed the study.

Intervention: All nursing staff on the intervention units received a group teaching package to improve their clinical practice in patient positioning.

Main outcome measure: Rivermead Mobility Index (RMI) at six months post stroke. Patient's position was recorded using an established observational tool.

Results: After the teaching there was some evidence of better positioning in the intervention than the control group (difference in percentage of correct positions per patient 4.9%, 95% confidence interval (CI-0.1% to 9.9%, p-0.055). There was no evidence of differences between the two groups in any of the outcome measures at six months although there was a trend towards increased elbow flexor tone in the control group.

Conclusions: A teaching intervention to improve patient positioning made no significant impact on outcome at six months post stroke. However, following the teaching there was only a slightly higher incidence of recommended patient positioning within the intervention group. Thus, a teaching package may not be powerful enough to enable any effect on patient outcome to be measured.
Release of Transgenic Human Insulin from Gastric G Cells: A Novel Approach for the Amelioration of Diabetes
Authors: Lu Yu-Chun , Sternini Catia , Rozengurt Enrique , Zhukova Elena
Published: February 2005

We explored the hypothesis that meal-regulated release of insulin from gastric G cells can be used for gene therapy for diabetes. We generated transgenic mice in which the coding sequence of human insulin has been knocked into the mouse gastrin gene. Insulin was localized specifically to antral G cells of G-InsKi mice by double immunofluorescence staining using antibodies against insulin and gastrin. Insulin extracted from antral stomach of G-InsKi mice decreased blood glucose upon injection into streptozotocin-diabetic mice. Intragastric administration of peptone, a known potent luminal stimulant of gastrin secretion, induced an increase in circulating levels of transgenic human insulin from 10.7 ± 2 to 23.3 ± 4 pm in G-InsKi mice. Although G cell-produced insulin decreased blood glucose in G-InsKi mice, it did not cause toxic hypoglycemia. Proton pump inhibitors, pharmacological agents that increase gastrin output, caused a further increase in the circulating levels of gastric insulin (41.5 ± 2 pm). G cell-produced insulin was released into circulation in response to the same meal-associated stimuli that control release of gastrin. The most striking aspect of the results presented here is that in the presence of the G-InsKi allele, Ins2Akita/+ mice exhibited a marked prolongation of life span. These results imply that G cell-derived transgenic insulin is beneficial in the amelioration of diabetes. We suggest that an efficient G cells-based insulin gene therapy can relieve diabetic patients from daily insulin injections and protect them from complications of insulin insufficiency while avoiding episodes of toxic hypoglycemia.
Development, Maintenance, and Function of the Adrenal Gland in Early Postnatal Proopiomelanocortin-Null Mutant Mice
Authors: Karpac Jason , Ostwald Dirk , Bui Stephanie , Hunnewell Peggy , Shankar Malini , Hochgeschwender Ute
Published: February 2005

Adult mouse mutants homozygous for an engineered proopiomelanocortin (POMC)-null allele lack macroscopically distinct adrenal glands and circulating adrenal hormones. To understand the basis for this adrenal defect, we compared the development of adrenal primordia in POMC-null mice and littermate controls. POMC-null mutant mice are born with adrenal glands that are morphologically indistinguishable from those of their wild-type littermates. However, in mutants adrenal cells fail to proliferate postnatally and adrenals atrophy until they have disappeared macroscopically in the adult. While present, mutant adrenals are differentiated as evidenced by the presence of enzymes for the final steps in the synthesis of corticosterone, aldosterone, and catecholamines. However, in contrast to adrenals of wild-type littermates, adrenals of POMC-null mutants do not produce corticosterone, not even in response to acute stimulation with exogenous ACTH. They do produce aldosterone; however, it is produced at reduced levels correlating with adrenal size. Transplantation of POMC-null mutant adrenals to adrenalectomized wild-type littermates results in adrenals with normal morphology and production of both corticosterone and aldosterone. These findings demonstrate that POMC peptides are not required for prenatal adrenal development and that POMC peptides in addition to ACTH are required for postnatal proliferation and maintenance of adrenal structures capable of producing both glucocorticoids and mineralocorticoids.
Estrogen Therapy Fails to Alter Amyloid Deposition in the PDAPP Model of Alzheimer’s Disease
Authors: Green Pattie S. , Bales Kelly , Paul Steven , Bu Guojun
Published: February 2005

Epidemiological studies implicate estrogen deprivation as a risk factor for Alzheimer’s disease and postmenopausal estrogen replacement as protective factor. One potential mechanism involves estrogen attenuation of β-amyloid (Aβ) peptide accumulation. We examined the effect of estrogen on amyloid accumulation in female PDAPP mice, which express human amyloid precursor protein (APP) with the V717F mutation. These animals deposit Aβ 1–42 in the hippocampus and neocortex and develop Alzheimer-like neuropathology. Mice were subjected to ovariectomy, ovariectomy with estrogen replacement, or sham surgery at 3 months of age, and levels of cerebral Aβ 1–40 and 1–42 were determined after 5 months of treatment. Neither estrogen deprivation nor estrogen replacement altered Aβ accumulation in the hippocampus or neocortex. Similarly, immunoreactivity for full-length human APP and secreted APPα was unchanged. Estrogen status of the animals was confirmed using a variety of techniques, including uterine and pituitary weight, vaginal cytology, and plasma estradiol concentrations. There was no correlation between plasma estradiol levels and accumulation of either Aβ 1–40 or Aβ 1–42 in the brain. Our observations indicate that long-term estrogen therapy does not alter amyloid pathology in PDAPP mice, an animal model of Alzheimer’s disease, and question the role of estrogen in Aβ deposition in brain.
Congenital Adrenal Hyperplasia: The Molecular Basis of 21-Hydroxylase Deficiency in H-2aw18 Mice
Authors: Riepe Felix G. , Tatzel Stephan , Sippell Wolfgang G. , Pleiss Jürgen , Krone Nils
Published: February 2005

The mouse strain H-2aw18 shows typical characteristics of 21-hydroxylase deficiency (21-OHD). A deletion of the active Cyp21a1 gene has been postulated; however, the changes on the nucleotide level are still unknown. To investigate whether this animal model, the only one available, is suitable for studying congenital adrenal hyperplasia in man, a detailed analysis of the Cyp21 locus has been performed to ascertain the genetic cause of 21-OHD in H-2aw18 mice. We demonstrate that 21-OHD is caused by unequal crossing over between the active Cyp21a1 gene and the pseudogene resulting in a hybrid Cyp21a1-Cyp21a2-p gene including a partial deletion of Cyp21a1. Next to several pseudogene-specific point mutations, various novel missense mutations and a nonsense mutation are present. Enzyme activity for each point mutation has been determined in vitro and the structure-function relationship has been studied by sequence conservation analysis and a three-dimensional murine 21-hydroxylase protein (Cyp21) structure model. The mutations are classified in three classes: I, no or minor decrease in enzyme activity: R238Q, P465L, R361K, A362V, P458L; II, loss of enzyme activity caused by inefficient electron flux: R346H, R400C; III, loss of activity due to deficient substrate binding: I462F, L464F. The combination of in vitro protein expression and three-dimensional structure modeling provides a valuable tool to understand the role of the different mutations and polymorphisms on the resulting enzyme activity. The underlying genetic mechanisms are also known to be responsible for 21-OHD in humans, so rodent 21-OHD turns out to be an excellent genetic model for studying the human disease.
Blockade of Rapid Versus Prolonged Extracellularly Regulated Kinase 1/2 Activation Has Differential Effects on Insulin-Induced Gene Expression
Authors: Keeton Adam B. , Bortoff Katherine D. , Franklin J. Lee , Messina Joseph L.
Published: February 2005

In the present work, insulin’s regulation of expression of activating transcription factor 3 (ATF-3), the putative transcription factor proline-rich induced protein (Pip)92, and insulin-inducible gene-1 (Insig-1) (an ER resident protein involved in regulation of sterol-responsive element-binding protein 1 activation) have been examined in a liver-derived cell line (rat H4IIE hepatoma cells). We report that: 1) insulin-induced transcription of ATF-3, Pip92, and Insig-1 required MEK-ERK activation; 2) insulin-induced transcription of ATF-3 and Pip92 reached maximum levels within 15 min and was blocked by wortmannin but not LY294002; 3) in contrast, the maximum level of insulin-induced transcription of Insig-1 was delayed and was not blocked by either wortmannin or LY294002; 4) insulin activated ERK1/2 in two distinct phases, a rapid peak and a later plateau; 5) the delayed plateau phase of insulin-induced ERK1/2 activation was partially phosphatidylinositol 3-OH-kinase dependent; and 6) however, the rapid, insulin-induced peak of ERK1/2 activation was blocked by wortmannin but not LY294002.
Hydrothermal Synthesis of Highly Crystallized Lepidocrocite Nanosheets of TiO2 under Low Temperature
Authors: Wang Fumin , Jiu Jinting , Pei Lihua , Nakagawa Keizo , Isoda Seiji , Adachi Motonari
Published: February 2005

The highly crystallized lepidocrocite nanosheets of TiO2 was synthesized by hydrothermal methods at low temperature by using dodecanediamine (DDA) as a surfactant. The above structural features of the titanium oxide nanosheets were confirmed by transmission electron microscopy (TEM), selected area electron diffraction (SAED), X-ray diffraction (XRD) and UV–vis optical spectra analysis.
Strategic Pricing and Detailing Behavior in International Markets
Authors: Chintagunta Pradeep K. , Desiraju Ramarao
Published: February 2005

We study three determinants of the levels of price and detailing effort across geographic markets: the within-market response to each variable, the nature of interfirm strategic interactions both within that market and across markets. We empirically examine the interactions of Prozac, Zoloft, and Paxil across the United States, the United Kingdom, Germany, France, and Italy. Our results indicate that all three factors driving marketing mix interactions are at play in this product category. The U.S. market is less price-sensitive than the European markets. Detailing elasticities are comparable across the United States, Germany, and Italy, while the United Kingdom and France show greater elasticity. For the U.S. market, we find that almost all deviations from Nash pricing and detailing levels are due to within-market interactions. In the U.K. market, deviations from Nash prices come about due mostly to across-market interactions—both with the United States as well as the rest of Europe, whereas deviations from Nash detailing levels are mainly due to across-market interactions with the United States. For Italy, we observe that both within and across-market interactions affect price and detailing levels. Overall, the pattern of interactions makes observed prices more similar across countries than prices implied by the estimated elasticities. This underscores the importance of considering within- and across-market interactions in developing multimarket strategy.
Technological Discontinuities and Complementary Assets: A Longitudinal Study of Industry and Firm Performance
Authors: Rothaermel Frank T. , Hill Charles W. L.
Published: February 2005

We suggest that the type of complementary assets (generic versus specialized) needed to commercialize a new technology is critical in determining the industry- and firm-level performance implications of a competence-destroying technological discontinuity. At the industry level, we hypothesize that incumbent industry performance declines if the new technology can be commercialized through generic complementary assets, whereas incumbent industry performance improves if the new technology can be commercialized through specialized complementary assets. At the firm level, we posit that an incumbent firm's financial strength has a stronger positive impact on firm performance in the postdiscontinuity time period if the new technology can be commercialized through generic complementary assets. We hypothesize, however, that an incumbent firm's R&D capability has a stronger positive impact on firm performance in the postdiscontinuity time period if the new technology can be commercialized through specialized complementary assets. Drawing on multi-industry, time series, and panel data over a 26-year period to analyze pre- and postdiscontinuity industry and firm performance, we find broad support for our theoretical model.
Scintillation Proximity Assay
Authors: Kahl Steven D. , Felder Christian C.
Published: February 2005

Scintillation proximity assay technologies provide a rapid non‐separation method to measure common biological interactions using radioactively tagged molecules. This unit identifies potential uses of the technology for the measurement of receptor‐ligand binding, cAMP accumulation, GTP binding to heterotrimeric G proteins, protease activity and cellular uptake.
Defensive Responses to Predator Threat in the Rat and Mouse
Authors: Blanchard D. Caroline , Blanchard Robert J. , Griebel Guy
Published: February 2005

Defensive responses include an array of specific behaviors, including flight, freezing, risk assessment, and defensive threat/attack, that are elicited by unconditioned threat stimuli such as predators or predator odors. Some individual defensive behaviors are selectively responsive to drugs effective against generalized anxiety disorder or panic, providing a rationale for their use in investigation of compounds that may be useful in treating these disorders. In addition, defensive behaviors toward predators and some predator odors show rapid conditioning to contextual stimuli, whereas other predator odors do not, although they too elicit defensiveness. This pattern suggests that the ability of a predator odor to predict danger may be a determinant of the degree to which that odor supports aversive conditioning. Predators and predator odors are also increasingly used in studies of brain systems potentially related to emotionality. These factors indicate the need for selective, reliable, and convenient tests of defensiveness to predators and predator odors using rat and mouse subjects.
Difference Gel Electrophoresis (DIGE) Using CyDye DIGE Fluor Minimal Dyes
Authors: Chakravarti Bulbul , Gallagher Sean R. , Chakravarti Deb N.
Published: February 2005

One‐ and two‐dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis (1‐ and 2‐D SDS‐PAGE) have been widely used for the separation and quantitative estimation of proteins. Following electrophoresis, the gels are stained appropriately to visualize the proteins. Difference gel electrophoresis (DIGE) is a new technique in which different protein samples, individually labeled with specific CyDyes, are combined together followed by electrophoresis and post electrophoretic co‐detection and co‐analysis on the same gel. CyDye DIGE fluor minimal dyes, which consist of three different CyDyes with different spectral characteristics, have been widely used for such purposes. The technique is highly sensitive with a wide dynamic range for detection of proteins and compatible with state‐of‐the‐art protein identification techniques using mass spectrometry. Although DIGE is mainly used to compare differential expression of various protein samples using 2‐D SDS‐PAGE, 1‐D DIGE also has important applications in quantitative proteomic studies.
Micrococcal Nuclease Analysis of Chromatin Structure
Authors: Zaret Ken
Published: February 2005

This unit describes methodology for using micrococcal nuclease to investigate the presence of nucleosomes at a particular location in chromatin and to map the positions of nucleosomes at various levels of resolution. The approaches are readily adaptable to other probes of chromatin structure that cause DNA cleavage. Results obtained from such chromatin studies provide a structural view of the molecular environment of gene in their native context in cells.
Chromatin Immunoprecipitation for Determining the Association of Proteins with Specific Genomic Sequences In Vivo
Authors: Aparicio Oscar , Geisberg Joseph V. , Sekinger Edward , Yang Annie , Moqtaderi Zarmik , Struhl Kevin
Published: February 2005

Chromatin immunoprecipitation (ChIP) is a powerful and widely applied technique for detecting the association of individual proteins with specific genomic regions in vivo. Live cells are treated with formaldehyde to generate protein‐protein and protein‐DNA cross‐links between molecules that are in close proximity on the chromatin template in vivo. DNA sequences that cross‐link with a given protein are selectively enriched, and reversal of the formaldehyde cross‐linking permits recovery and quantitative analysis of the immunoprecipitated DNA. As formaldehyde inactivates cellular enzymes essentially immediately upon addition to cells, ChIP provides snapshots of protein‐protein and protein‐DNA interactions at a particular time point, and hence is useful for kinetic analysis of events occurring on chromosomal sequences in vivo. In addition, ChIP can be combined with microarray technology to identify the location of specific proteins on a genome‐wide basis. in this unit describes the ChIP procedure for Saccharomyces cerevisiae; describes the corresponding steps for mammalian cells.
Pattern Discovery in Expression Profiling Data
Authors: Katagiri Fumiaki , Glazebrook Jane
Published: February 2005

In expression profiling studies, it is often necessary to identify groups of genes with similar expression profiles in a variety of samples, and/or groups of samples with similar expression profiles. Each profile can be expressed as a single data point in a space with the same number of dimensions as there are parameters in the profiles. In this way, pattern discovery among expression profiles is translated into pattern discovery in the spatial distribution of data points. Hierarchical clustering is useful for clustering similarly behaving genes or samples at local levels and for displaying the results in a simple color‐coded manner. K‐means clustering can be used for discovery of well‐defined clusters. Principal component analysis and self‐organizing maps can be used for dimensionality reduction, thereby facilitating visualization of major trends in data sets.
High‐Resolution FISH Analysis
Authors: Heng Henry H.Q. , Windle Bradford , Tsui Lap‐Chee
Published: February 2005

Map order, orientation, and gap or overlap distance of closely linked DNA probes may be determined using fluorescent hybridization to decondensed DNA. The linear arrangement of released chromatin fibers not only simplifies the task of gene ordering, but also provides higher resolution with probes separated by greater distances than can be achieved in FISH with intact interphase nuclei. Theof this unit describes an alkaline lysis procedure for generating free chromatin from cultured cells for FISH analysis. A support protocol describes an empirical approach to optimize conditions for preparation of free chromatin. Anprovides a method for producing free chromatin from cultured lymphocytes with drug treatment. The, high‐resolution FISH mapping with free chromatin, is a modification of the method used for FISH mapping of interphase nuclei.
Morphology Antibody Chromosome Technique for Determining Phenotype and Genotype of the Same Cell
Authors: Knuutila Sakari
Published: February 2005

The morphology antibody chromosome (MAC) technique is a powerful combination of methods that permits analysis of both phenotype and genotype on a single interphase or mitotic cell as a basis for lineage analysis of neoplastic and normal cells. This unit describes MAC with sequential phenotypic analysis using antibody and an alkaline phosphatase anti‐alkaline phosphatase (APAAP) complex and genotypic analysis using in situ hybridization with either enzymatic or fluorescence detection. Alternate methods for phenotypic analysis are also described, which include the use of horseradish peroxidase‐conjugated antibodies, fluorochrome‐conjugated antibodies, May‐Grunwald‐Giemsa cytochemical staining, and Sudan black B cytochemical staining. An additional protocol describes G‐ and C‐banding as alternatives to in situ hybridization (ISH) for genotyping MAC specimens. Support protocols describe methods for preparing specimenscytospin preparations, in situ cultures, paraffin‐embedded or cryostat sections, and blood and bone marrow smears. describes a procedure for chromosome painting of previously GTG‐banded slides.
Gene Expression Analysis of a Single or Few Cells
Authors: Klein Christoph A. , Zohlnhöfer Dietlind , Petat‐Dutter Karina , Wendler Nicole
Published: February 2005

The need to analyze rare cells is based on the nature of tissue differentiation and regeneration, the initiation and propagation of disease processes in multicellular organisms, and the functional diversity of individual cells. Gene transcription is the most important regulatory mechanism by which a phenotype and functional state of a cell is determined. Therefore, procedures for the qualitative and quantitative assessment of mRNA abundance are important. This unit presents a protocol for semi‐quantitative analysis of gene expression of a single cell and quantitative representation of expressed genes from >10 to 30 cells. A basic protocol for array hybridization on nylon filters is provided because such filters are available in every laboratory. Tissue samples contain many different cell types in variable amounts, so their analysis may require microdissection; a protocol for obtaining cryosections is given. Finally, a simple procedure to prepare the data for statistical analysis is also provided.
Comparative Overview of Flow and Image Cytometry
Authors: Robinson J. Paul
Published: February 2005

This unit considers the issues of which tool to use—flow cytometry or imaging—and under what conditions. In particular, it compares the advantages and disadvantages of flow and image cytometry and provides examples illustrating the proper choice of each technology. The result is a better understanding of why the two technologies are complementary in many applications. It is clear that many scientists use the tools that are familiar to them, often in preference to the best tool. In cases where very advanced and rather expensive technologies are concerned, this is not surprising. However, there are clearly times when one form of cytometry is definitely superior to another. What then constitute the criteria for a decision when both flow cytometry and imaging are available? This unit addresses some of these concerns.
Modern Confocal Microscopy
Authors: Rajwa Bartek
Published: February 2005

This unit re‐examines confocal microscopy from a current perspective. It outlines many of the most modern applications of confocal microscopy and the issues surrounding them. The expanding applications of confocal microscopy demand both minor and major modifications of the technology to enhance imaging capabilities for a growing variety of samples. Techniques of interest such as FRET, FLIP, FRAP, and IFRAP are described. The unit includes a discussion on multispectral imaging, potentially the most exciting innovation in the field of biological imaging. The other area that is beginning to have considerable impact on biology, medicine, and pharmacology comprises high‐content screening (HCS) and high‐throughput screening (HTS). This unit also introduces programmable array microscopes (PAMs), which are based upon a technique in which spatial modulators are placed in the imaging plane of a microscope and used to generate patterns of illumination and/or detection.
Titelbild: Verknoten und Durchfädeln von Molekülen: Chemie und Chiralität molekularer Knoten und ihrer Ensembles (Angew. Chem. 10/2005)
Authors: Lukin Oleg , Vögtle Fritz
Published: February 2005

Fantasiestrukturen wie der im Titel gezeigte siebenfache Kleeblattknoten, der mit sieben gewöhnlichen Kleeblattknoten verschränkt ist, sind weit entfernte Syntheseziele. Bewährte Templatstrategien zum Aufbau molekularer Knoten (Knotane) und höherer Ensembles mit verschlungenen Architekturen könnten einmal zu solchen Gebilden führen. Mehr über die Synthese, Funktionalisierung und Eigenschaften von Knotanen erfahren Sie im Aufsatz von F. Vögtle und O. Lukin auf S. 1480 ff.
Accuracy verification of spaceborne radar estimates of rain rate
Authors: Amitai E. , Liao L. , Llort X. , Meneghini R.
Published: February 2005

The distribution of rain rate is of great concern for many hydrological applications. Probability distribution functions (pdf) of rain rate can now be obtained from spaceborne radar observations. Effort to evaluate these pdfs using ground observations is described. Copyright © 2005 Royal Meteorological Society
Very short period quantitative precipitation forecasting
Authors: Fox Neil I. , Wilson James W.
Published: February 2005

This article presents an overview of the state of the art of very short period quantitative precipitation forecasting. The authors draw primarily on work presented during the sessions on ‘Nowcasting’ held at the 6th Symposium on the Hydrological Applications of Weather Radar, in Melbourne, Australia, from 2nd to 4th February 2004, and also include some other work in order to give a more complete picture of the field. Copyright © 2005 Royal Meteorological Society
Combining radar and rain gauge rainfall estimates using conditional merging
Authors: Sinclair Scott , Pegram Geoff
Published: February 2005

The Hydrologist's traditional tool for measuring rainfall is the rain gauge. Rain gauges are relatively cheap, easy to maintain and provide a direct and suitably accurate estimate of rainfall at a point. What rain gauges fail to capture well is the spatial variability of rainfall with time, an important aspect for the credible modelling of a catchment's response to rainfall. This spatial variability is particularly evident at short timescales of up to several days. As the period of accumulation increases, the expected spatial variability is reduced and rain gauges provide improved spatial rainfall estimates. Because of the fractal variability of rainfall in space, simple interpolation between rain gauges does not provide an accurate estimate of the true spatial rainfall field, at short time scales.

Weather radar provides (with a single instrument) a highly detailed representation of the spatial structure and temporal evolution of rainfall over a large area. Estimated rainfall rates are derived indirectly from measurements of reflectivity and are therefore subject to a combination of systematic and random errors.

This article describes a recently proposed merging technique and presents an application to simulated rainfall fields. The technique employed is Conditional Merging (Ehret, 2002), which makes use of Kriging to extract the optimal information content from the observed data. A mean field based on the Kriged rain gauge data is adopted, while the spatial detail from the radar is retained, reducing bias, but keeping the spatial variability observed by the radar. The variance of the estimate is reduced in the vicinity of the gauges where they are able to provide good information on the true rainfall field. Copyright © 2005 Royal Meteorological Society
Correction of the bright band using dual‐polarisation radar
Authors: Rico‐Ramirez M. A. , Cluckie I. D. , Han D.
Published: February 2005

This article presents an algorithm to correct the increase in reflectivity in the melting layer using weather radars. The correction starts with a fuzzy logic system (FLS) to classify rain, snow and melting snow using the reflectivity factor, the differential reflectivity, the linear depolarisation ratio and the height of the measurement from the ground surface. Using the output of the FLS, a correction is performed on the bright band contaminated reflectivity data assuming an idealised vertical reflectivity profile typical from stratiform precipitation and obtained from a considerable number of observations at S‐band frequencies. Copyright © 2005 Royal Meteorological Society
Assessing error in hydrological and hydraulic model output flows
Authors: Robbins Gavin L. , Collier Chris. G.
Published: February 2005

A hydraulic model of an urban drainage system with a Bayesian post processor is used to assess the uncertainty in model output flows associated with the use of measurements of rainfall made using raingauges, microwave link attenuation measurements and weather radar. Copyright © 2005 Royal Meteorological Society
Colour remediation of pulp mill effluent using purified fungal cellobiose dehydrogenase: Reaction optimisation and mechanism of degradation
Authors: Wingate Kathryn G. , Stuthridge Trevor , Mansfield Shawn D.
Published: February 2005

Cellobiose dehydrogenase purified from two different fungal sources was assessed for its ability to remove and/or reduce colour from pulp mill bleach plant effluent. Cellobiose dehydrogenase purified from Phanerochaete chrysosporium was shown to prefer acidic conditions and was consequently used to treat the acid effluent stream discharged from a pulp mill bleach plant, while an analogous enzyme originating from Humicola insolens preferred alkaline conditions, and was applied to the effluent discharged from the caustic sewer of the bleach plant. Both enzyme preparations were able to remove colour from their respective effluent sources to a comparable extent. Up to 50% of the effluent colour was removed within 4 days when treated under optimised conditions. Furthermore, it was also shown that this enzymatic approach was effective at removing colour generated by both softwood and hardwood resources. Mechanistically, it was shown that colour was removed from all molecular weight fractions, and the higher molecular weight material (>300 kDa) was concurrently preferentially degraded. Cellobiose dehydrogenase treatment of effluent did not target phenolic, stilbene, or α‐carbonyl structures, but did affect the quinone content. Further investigations using model compounds confirmed these results, and subsequently showed that only the para‐quinones with low substitution were reduced with cellobiose dehydrogenase. © 2005 Wiley Periodicals, Inc.