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Efficient allelic exchange and transposon mutagenesis in Mycobacterium avium by specialized transduction.

Mycobacterium tuberculosis and Mycobacterium avium are pathogenic slow-growing mycobacteria that cause distinct human diseases. In contrast to recent advances in M. tuberculosis genetics and pathogenesis investigation, M. avium has remained genetically intractable and, consequently, its pathogenic strategies remain poorly understood. Here we report the successful development of efficient allelic exchange and transposon mutagenesis in an opaque clinical strain of M. avium by specialized transduction. Efforts to disrupt the leuD gene of M. avium by specialized transduction were successful but were complicated by inefficient isolation of recombinants secondary to high spontaneous antibiotic resistance. However, by using this leucine auxotroph as a genetic host and the Streptomyces coelicolor leuD gene as a selectable marker, we achieved efficient allelic exchange at the M. avium pcaA locus. A leuD-marked transposon delivered by specialized transduction mutagenized M. avium with efficiencies similar to M. tuberculosis. These results establish a system for random and directed mutagenesis of M. avium. In combination with the forthcoming M. avium genome sequence, these tools will allow the distinct physiologic and pathogenic properties of M. avium to be dissected in molecular detail.

Alleles↗

Patterns of admissions for children with special needs to the paediatric assessment unit.

BACKGROUND: Children with special needs present a challenge to those involved in their care. AIMS: To determine the role of the acute assessment unit for these children. METHODS: Case notes and other records were reviewed for information on referrals, admissions, readmission within 7 and 28 days, length of stay, and management of 86 children registered for special needs. The study covered five years between January 1997 and December 2001. RESULTS: Of the 86 children, 48 (58%) were boys; 62 children had cerebral palsy and 52 learning disability. There were 914 episodes, with 44% of these being self referrals and 35% from general practitioners; 35.5% of the episodes were managed in the assessment unit. The average length of stay in hospital was 5 days, ranging from <24 hours to 63 days; 37.5% of those admitted to the ward stayed for less than 24 hours. Respiratory tract infections and seizures were the main reasons for referral and admission. CONCLUSION: Children with special needs tend to have a predictable pattern of conditions requiring inpatient care. One third of the inpatients episodes did not need a prolonged stay in hospital. This latter group of children could be managed at home with support of community nurses. Integrated care pathways need to be developed to minimise disruption to their lives. Appropriate resources should be made available to achieve these goals.

Adolescent↗

Contact lenses and special back surface design after penetrating keratoplasty to improve contact lens fit and visual outcome.

AIMS: To describe the fitting of patients with high or irregular astigmatism following penetrating keratoplasty with contact lenses and to answer the question whether or not contact lenses with special back surface design can improve visual acuity in complex cases after penetrating keratoplasty. METHODS: 28 eyes were included. They were fitted with contact lenses with a special back surface that was designed for optical rehabilitation after penetrating keratoplasty. Four different types of these lenses (tricurve, keratoconus, reverse, oblong) were used selectively depending on abnormal eccentricity determined by videokeratoscope. The patients were followed up for an average period of 15.5 months. Lens tolerance and corrected visual acuity were evaluated and compared with that corrected with spectacles. RESULTS: The visual acuity was significantly improved in nearly all eyes with an average increase of 3.6 lines (maximal nine lines) accompanied by good contact lens tolerance and satisfactory contact lens fit. No noticeable complications were observed. CONCLUSION: Contact lenses with special back surface design can improve visual results and lens tolerance, and minimise problems in contact lens fitting. This is in favour of contact lenses as an alternative to surgical procedures for correction of high or irregular astigmatism after penetrating keratoplasty. This procedure is recommended especially in cases of patients who decline further operative interventions.

Adult↗

Arrangements for special and intensive care of the newborn.

All the special care baby units in three Thames health regions completed a one-day census on the number of infants in the unit, staffing, and facilities. The results were assessed in the light of the recommendations of an expert group set up by the Department of Health and Social Security. Although the provision of cots (6.1-6.9 per 1000 live births) was more generous than the estimated need and the occupancy rate (59--63%) lower than suggested, some units did not have adequate trained nursing cover at night, some did not have resident paediatricians, and some lacked essential equipment. Some small units, on the other hand, had very expensive equipment that was probably underused. Relating this provision to the regional statistics on babies needing special care showed that there was room for the rationalisation of facilities for their care. Factors that should be considered in planning such care include the size of the child-bearing population, the percentage of infants of low birth weights, and the proximity of specialist paediatric and maternity units and pathology facilities. It is also essential to monitor the effectiveness of the care by assessing the outcome in babies admitted to special units, especially those of low birth weight.

Health Facility Planning↗

Special care nurseries: admitting to a policy.

Few criteria exist for identifying which babies merit special nursing care, and many asymptomatic babies are offered such care unnecessarily. Birth weight is usually used to identify babies in need of special care at birth, but gestational age is a better discriminator. Babies of more than 35 weeks' gestation do not normally require admission to a special care nursery. Those preterm babies who are admitted can usually be discharged at 37 weeks' gestation even if they still weigh less than 2000 g.

Bed Occupancy↗

The limited potential of special ambulance services in the management of cardiac arrest.

For six months a survey was made of all the patients in the Nottingham District Health Authority who died or who were brought to hospital after a cardiac arrest outside hospital. During this period just under half of the emergency ambulance shifts were covered by specially trained crews with defibrillators. During the study period the ICD coding of death certificates indicated that 894 (25%) of the 3575 deaths were due to ischaemic heart disease. During this period the ambulance service received 17,749 emergency calls, which included 445 patients who had cardiac arrests outside hospital. One hundred and forty seven of these patients were carried by ambulances equipped with defibrillators and resuscitation was attempted in 83. Seven patients survived to leave hospital. The special ambulance service was cost effective--a simple calculation suggests that the cost per life saved was approximately 2600 pounds, but it seems unlikely that special ambulance services will materially affect community fatality rates from ischaemic heart disease.

Adult↗

Doctors, ethics and special education.

This discussion paper is drawn from a qualitative research project comparing the effect of special and ordinary schools on the lives of children, young people and their families. Special schools are recommended by health professionals who seldom know how ineffective these schools are. We question the beneficence and justice of health professionals' advice on education for children with disabilities and other difficulties. Cooperation with local education authorities (LEAs) plays a considerable part in the work of community paediatricians, clinical medical officers, therapists and other health professionals encountering children with "special needs". The "needs" range from physical disability and sensory impairment to learning difficulties and emotional or behavioural difficulties. This cooperation involves routine administrative problems, but it raises broad ethical issues too, particularly in respect of current tendencies in state schooling towards the integration or inclusion of these children in mainstream schools and classes.

Adolescent↗

MR imaging using specialized coils.

The purpose of this study was to evaluate the clinical utility of specialized coils with magnetic resonance (MR) imaging and to determine the potential improvement in the signal-to-noise ratio (SNR) provided by the various coils. A total of 48 patients were studied-33 with surface coils and 15 with limb coils. MR imaging was performed with a superconducting magnet operating at 0.35 T. Two applications of surface coils were evaluated; one used for both transmitting and receiving and the other used only for receiving. Structures studied with the surface coils included the orbit, carotid artery, cervical and lumbar spine, parotid and thyroid glands, other neck structures, femoral artery, kidney, prostate, hip, and heart. The regions studied with the limb coil were the ankle, knee, wrist, elbow and popliteal artery. We calculated the SNR, and the results demonstrated a marked improvement with the specialized coils: a twofold increase for the 10-cm surface coil versus the head coil and a 4.6-fold increase compared with the body coil, a 2.3-fold increase for the 20-cm surface coil compared with the body coil, and 2.8- and 6.4-fold increases for the limb coil versus the head coil and the body coil, respectively. The use of specialized coils was feasible in the clinical setting and they provided a marked improvement in the image SNR, which facilitated the imaging of small and superficial structures.

Adolescent↗

Measurements of middle-ear function in the Mongolian gerbil, a specialized mammalian ear.

The middle ear of the Mongolian gerbil is thought to be specialized so that it can hear the 3,000-Hz and lower sounds produced by approaching predators. Evidence in the literature suggests several specializations: (1) an effect of the large middle-ear air spaces in the gerbil is to increase the sensitivity of this ear to sounds of 3, 000 Hz and lower in frequency; (2) the combined action of the middle-ear cavity and the significant pars flaccida of the tympanic membrane in the gerbil ear reduces the sensitivity to sounds of frequencies below 500 Hz. This paper describes anatomical and functional measurements of stapes motion and suggests that the stiffness of the gerbil ossicular system also acts to reduce sensitivity to low-frequency sounds. The primary conclusion is that the frequency dependence of the gerbil middle ear is the result of the interaction of multiple factors, thereby complicating the relationship between single structural specializations and overall function.

Acoustic Stimulation↗

Radiation and functional specialization of the family-3 glycoside hydrolases.

A phylogenetic analysis of the glycoside hydrolases of family 3 (GH3s) was conducted in order to infer particular trends in its evolution: functional specialization, gene transfer events, gene duplications and paralogous evolution, and gene deletions. The phylogenetic analysis of GH3s revealed six clusters, i.e., A, B, C, D, E, and F that could fit the definition of 3 sub-families, i.e., AB, AB' and AB". While the sub-families AB' and AB" contain a single cluster, F and E, respectively, the AB sub-family is sub-divided into four clusters. Global analysis of the GH3 phylogenetic tree suggests a primary burst of amplification of the GH3s that might have led to these sub-families. Specializations, gene transfers, and gene duplications among each of these sub-families and phylogenetic clusters might then have occurred and have been inferred. The fine comparison of the enzyme properties and phylogenetic relationships of GH3s allowed to detect common functional groups that belong to the same cluster (D, E or F), or sub-cluster (A1, A2 or B2). The prokaryotic and eukaryotic beta-xylosidases and beta-glucosidases belong to the AB and AB' sub-families, and the N-acetylglucosaminidases are in sub-family AB" (in cluster E). In some instances (B1, B2, C1, C2, and C3), the lack of data and/or the high heterogeneity of the hydrolytic properties did not allow to infer a particular link between an enzyme functional group and a phylogenetic cluster, suggesting the emergence of some highly specialized GH3s.

Computational Biology↗

Memory and hippocampal specialization in food-storing birds: challenges for research on comparative cognition.

The three-way association among food-storing behavior, spatial memory, and hippocampal enlargement in some species of birds is widely cited as an example of a new 'cognitive ecology' or 'neuroecology.' Whether this relationship is as strong as it first appears and whether it might be evidence for an adaptive specialization of memory and hippocampus in food-storers have recently been the subject of some controversy [Bolhuis and Macphail, 2001; Macphail and Bolhuis, 2001]. These critiques are based on misconceptions about the nature of adaptive specializations in cognition, misconceptions about the uniformity of results to be expected from applying the comparative method to data from a wide range of species, and a narrow view of what kinds of cognitive adaptations are theoretically interesting. New analyses of why food-storers (black-capped chickadees, Poecile Atricapilla) respond preferentially to spatial over color cues when both are relevant in a memory task show that this reflects a relative superiority of spatial memory as compared to memory for color rather than exceptional spatial attention or spatial discrimination ability. New studies of chickadees from more or less harsh winter climates also support the adaptive specialization hypothesis and suggest that within-species comparisons may be especially valuable for unraveling details of the relationships among ecology, memory, and brain in food-storing species.

Animals↗

The homing pigeon hippocampus and space: in search of adaptive specialization.

The hippocampus (HF) of birds and mammals is essential for the map-like representation of environmental landmarks used for navigation. However, species with contrasting spatial behaviors and evolutionary histories are likely to display differences, or 'adaptive specializations', in HF organization reflective of those contrasts. In the search for HF specialization in homing pigeons, we are investigating the spatial response properties of isolated HF neurons and possible right-left HF differences in the representation of space. The most notable result from the recording work is that we have yet to find neurons in the homing pigeon HF that display spatial response properties similar to HF 'place cells' of rats. Of interest is the suggestion of neurons that show higher levels of activity when pigeons are near goal locations and neurons that show higher levels of activity when pigeons are in a holding area prior to be being placed in an experimental environment. In contrast to the rat, the homing pigeon HF appears to be functionally lateralized. Results from a current lesion study demonstrate that only the left HF is sensitive to landmarks that are located within the boundaries of an experimental environment, whereas the right HF is indifferent to such landmarks but sensitive to global environmental features (e.g., geometry) of the experimental space. The preliminary electrophysiological and lateralization results offer interesting departure points for better understanding possible HF specialization in homing pigeons. However, the pigeon and rat HF reside in different forebrain environments characterized by a wulst and neocortex, respectively. Differences in the forebrain organization of pigeons and rats, and birds and mammals in general, must be considered in making sense of possible species differences in how HF participates in the representation of space.

Animals↗

Prospective, comparative cohort studies and their contribution to the benefit assessments of therapeutic options: heart failure treatment with and without Hawthorn special extract WS 1442.

BACKGROUND: In addition to testing a drug for its efficacy, pharmacological quality and safety, current policies are increasingly demanding evaluations of the therapeutic benefits provided by a drug in general practice with "non-selected" patients and increasingly restrictive economic considerations. OBJECTIVE: One of the trials which addresses this task is the WISO cohort study (Efficacy and socio-economic relevance of treatment of chronic heart failure stage NYHA II with Crataegus extract WS 1442). It compares two different therapeutic strategies in the treatment of heart failure stage NYHA II, i.e. a conventional medication and a therapy which also includes hawthorn special extract WS 1442 (Crataegutt novo 450) in addition to chemical-synthetic drugs. In contrast to clinical trials, the patients in cohort studies are expressly not randomised and the physician in charge independently chooses the administered treatment. This comparative, non-interventional observation provides well-founded evidence of the "real-world effectiveness" of the tested preparation. PATIENTS AND METHODS: 952 patients with heart failure (NYHA II) were enrolled in the study by 217 general practitioners. 588 patients received Crataegus special extract WS 1442 (Crataegutt novo 450) either as an add-on therapy or as a monotherapy (Crataegus cohort) and 364 patients received therapy without hawthorn (comparative cohort). These two groups had the same indication (heart failure NYHA II) but were significantly different regarding gender, age and concomitant cardiovascular disease. Basically, in view of the free choice of therapy made by the physician in charge, such differences are to be expected in comparative observational studies. A sufficient degree of patient comparability was provided by means of the matched-pairs technique, which replaced the randomisation procedure normally used in clinical studies. After 2 years, 130 patient pairs generated by this technique could be included in the interim assessment. RESULTS: The clinical symptoms with regard to all parameters investigated showed the same or a more pronounced improvement in the Crataegus cohort in the course of 2 years. After 2 years, the three cardinal symptoms of heart failure--fatigue (p = 0.036), stress dyspnoea (p = 0.020) and palpitations (p = 0.048)--were significantly less marked in the Crataegus cohort than in the comparative cohort. DISCUSSION: The particular design of the cohort study also provides valuable additional information: (1) Hawthorn special extract WS 1442 was prescribed in registered cardiological practices for the treatment of patients with heart failure stage NYHA II, partly as an alternative and partly as a supplement to the used chemical-synthetic drugs. (2) Favourable effects on the clinical symptoms were achieved although the patients in the Crataegus cohort received markedly fewer chemical-synthetic drugs than the patients in the comparative cohort (ACE-inhibitors: 36 vs. 54%, p = 0.004; cardiac glycosides: 18 vs. 37%, p = 0.001; diuretics: 49 vs. 61%, p = 0.061; beta-blockers: 22 vs. 33%, p = 0.052). CONCLUSION: The data show a clear benefit for patients with heart failure stage NYHA II treated with WS 1442. The single or add-on administration in addition to a chemical-synthetic medication resulted in objective improvements at comparable costs.

Cohort Studies↗

Reports by some massage therapists of a sense of 'special connection' that is sometimes experienced with clients/patients.

BACKGROUND: The aim of this study was to provide exploratory qualitative data on anecdotal reports by some physical therapists that they sometimes experience a sense of 'special connection' with patients or clients. PARTICIPANTS AND METHODS: 21 self-selected massage therapists provided written comments about their experience of therapy in response to a questionnaire (free-form response format) asking about the experience of connection. RESULTS: 20 out of 21 therapists reported a sense of 'special connection'. Content analysis suggested 10 themes, the main ones relating to the special connection being: (a) a sense of oneness or unity with the patient; (b) intimacy, trust and the touching of souls; (c) meditative, trance-like, floating relaxation; (d) 2-way flow of energy, and the mutual contribution of patient and therapist characteristics in forming this connection; (e) that it tends to be there or not but if there, it varies in degree. CONCLUSION: These reports are similar in some respects to descriptions given by healers of the experience of healing, suggesting that some massage therapists, and possibly some other complementary or conventional therapists engage, unknowingly, in a process of healing while they practice the therapy for which they have been trained.

Female↗

Advantages and disadvantages of special depression treatment and research wards.

After the establishment of the first European depression research ward at the PUK Basle 1968 we started the Weissenau 'depression treatment and research ward' in 1976. Since this time there were approximately 1800 admissions of depressed inpatients. Up to this day 3 of the county mental hospitals of Baden-Württemberg FRG (1984 2797 depressive inpatients, average time of stay 49 days) established also such special treatment wards. The conceptualization of depression wards due to treatment strategies is still in the beginning and based on clinical experience up to now. Some advantages of such depression wards could be treating a homogeneous group of patients, the chance for special psychotherapeutic treatment strategies (reinforcement of non-depressive behaviour, empathic understanding of underlying dynamics), reduced anxiety in dealing with suicidal and clinging patients, a better dealing with regressive tendencies, or also better conditions for research programmes. Some of the disadvantages could be an uncontrollable regression under too empathic conditions or an accumulation of severely depressed which can paralyze therapeutic activity. To realize such a psychotherapeutic and pharmacological treatment and research program it is necessary to have staff members specialized and also supervised in dealing with depressed patients.

Combined Modality Therapy↗

Introduction of a flexible polymeric heart valve prosthesis with special design for mitral position.

BACKGROUND: Current heart valve prostheses are constructed mimicking the native aortic valve. Special hemodynamic characteristics of the mitral valve such as a nonaxial central inflow with creation of a left ventricular vortex have so far not been taken into account. A new polycarbonaturethane (PCU) bileaflet heart valve prosthesis with special design for the mitral position is introduced, and results of animal testing are presented. METHODS AND RESULTS: After in vitro testing, 7 PCU-prostheses and 7 commercial bioprostheses (Perimount, n=4; Mosaic, n=3) were implanted in mitral position into growing Jersey calves (age 3-5 months, weight 60-97 kg) for 20 weeks. 2-Dimensional echocardiography was performed after implantation and before sacrification. Autopsy included histologic, radiographic, and electron microscopic examination of the valves. In vitro durability was proven for >15 years. After implantation 2-dimensional-echocardiography showed no relevant gradient or regurgitation of any prosthesis. Clinical course of the animals with PCU valves was excellent. In contrast, 5 of 7 calves with bioprostheses were sacrificed after 1-9 weeks because of congestive heart failure. 2-Dimensional echocardiography of the PCU valves after 20 weeks showed mild leaflet thickening with trivial regurgitation; mean gradient was 8.1+/-5.0 mm Hg (weight: 160-170 kg). The explanted PCU prostheses revealed mild calcification and no structural degeneration. All of the Perimount bioprostheses were severely calcified and degenerated after 11+/-7 weeks. One Mosaic bioprosthesis was thrombosed after 1 week, and 2 showed severe and mild-to-moderate degeneration after 4 and 22 weeks, respectively. CONCLUSIONS: Polycarbonaturethane valve prostheses with special design for mitral position show excellent hemodynamic performance and durability in vivo. Calcification and structural changes are mild compared with bioprostheses. Controlled clinical studies are planned.

Animals↗

Electrophysiological delineation of the specialized A-V conduction system in patients with corrected transposition of the great vessels and ventricular septal defect.

The specialized atrioventricular (A-V) conduction system was electrophysiologically delineated during open-heart surgery in four patients with congenitally corrected transposition of the great vessels and associated ventricular septal defect and one patient with single ventricle. Two consistent observations were made: 1) In no case was the specialized A-V conduction system found in the right atrium, whether or not there was a coronary sinus ostium present. 2) Specialized A-V conduction system electrograms were never delineated posterior to the ventricular septal defect, in contradistinction to this usual location in hearts having ventricular septal defects associated with other congenital lesions. In three of five patients, the initial course of the A-V conduction system of the ventricles was delineated between the anterior aspect of the ventricular septal defect and the pulmonary artery. In one patient the proximal portion of the A-V conduction system was delineated on the anterior aspect of the pulmonary conus. The course and extent of the A-V conduction system delineated in the morphological left ventricle suggests it is a left bundle branch. The surgical implications of the ectopic location of the A-V conduction system anterior to the ventricular septal defect, and the variability of the more proximal portion of the A-V conduction system are discussed.

Adult↗

Intraoperative recording of specialized atrioventricular conduction tissue electrograms in 47 patients.

Intraoperative mapping of the specialized atrioventricular conduction system was performed in 47 patients during cardiac surgery. Specialized conduction tissue electrograms were identified in 37, and atrioventricular conduction preserved in 92%. Specialized conduction tissue was identified in 27 patients with atrioventricular canal defect: complete heart block was avoided in 25. Conduction tissue was located in six of 12 patients with complex transpositions; atrioventricular conduction was preserved in all six. Other lesions in which the technique was useful were Ebstein's anomaly and single atrium. Limitations to the technique are 1) deep hypothermia and circulatory arrest; 2) interruption in atrioventricular conduction during mapping; 3) inadequate exposure and access to probable sites of conduction tissue; 4) variation of size and spatial relations of individual malformations; and 5) limited time for identification of unusually located conduction tissue. Indications for use of this technique include patients with both forms of atrioventricular canal, complex transpositions, atrioventricular discordance, single ventricle and single atrium.

Adolescent↗