Integrating movement in academic classrooms: understanding, applying and advancing the knowledge base.

In the context of comprehensive and coordinated approaches to school health, academic classrooms have gained attention as a promising setting for increasing physical activity and reducing sedentary time among children. The aims of this paper are to review the rationale and knowledge base related to movement integration in academic classrooms, consider the practical applications of current knowledge to interventions and teacher education, and suggest directions for future research. Specifically, this paper (i) situates movement integration amid policy and research related to children's health and the school as a health-promoting environment; (ii) highlights the benefits of movement integration; (iii) summarizes movement integration programs and interventions; (iv) examines factors associated with classroom teachers' movement integration; (v) offers strategies for translating research to practice and (vi) forwards recommendations for future inquiry related to the effectiveness and sustainability of efforts to integrate movement into classroom routines. This paper provides a comprehensive resource for developing state-of-the-art initiatives to maximize children's movement in academic classrooms as a key strategy for important goals in both education and public health.

Evidence for the biosynthesis of a prolactin-releasing factor from the ovine pars tuberalis, which is distinct from thyrotropin-releasing hormone.

This study demonstrates the presence of two prolactin-releasing (PR) factors in media conditioned by primary pars tuberalis cells prepared from dispersed pars tuberalis tissue. One factor was identified as thyrotropin-releasing hormone (TRH) on the basis of immunoreactivity and following purification by high-performance liquid chromatography and mass spectrometry. The origin of TRH in the pars tuberalis conditioned media was investigated by measuring the expression of glutaminyl-cyclase (QC) by in situ hybridization. QC expression was not detected in pars tuberalis-specific cells, but was relatively abundant in cells in the pars distalis and hypothalamic paraventricular nucleus. These data suggest that TRH is not synthesized by the ovine pars tuberalis and more likely originated from the hypothalamic neuronal processes from the paraventricular nucleus that terminate in the median eminence. The second component of the conditioned media PR bioactivity was insensitive to the TRH-antiserum, less than 1 kDa and was not retained by the C18 reverse-phase column. The biosynthesis of the PR bioactivity by pars tuberalis cells was investigated using cycloheximide, forskolin and melatonin. Cycloheximide reduced the level of PR bioactivity produced by the pars tuberalis cells. Melatonin inhibited the increased level of PR bioactivity stimulated by forskolin. Collectively, these data demonstrate the synthesis of at least one regulator of prolactin secretion by ovine pars tuberalis-specific cells.

Use of RAPD-ALF analysis for investigating the frequency of bacterial cross-transmission in an adult intensive care unit.

Bacterial cross-transmission was investigated during a 12-month period in an adult intensive care unit (ICU) by the generation of random amplified polymorphic DNA (RAPD) fingerprinting profiles, combined with automated laser fluorescence (ALF) analysis. The potential episodes of cross-transmission identified, were compared with those detected by the conventional first-line screen of antibiogram typing. Over the year, 215 primary gram-negative bacterial isolates were obtained from 160 patients. In total, 22 possible episodes of cross-transmission, involving 70 (44%) of the 160 patients, were identified by RAPD-ALF analysis, and 19 of these were substantiated with epidemiological evidence. Conversely, 31 possible episodes were identified on the basis of antibiogram data, but only three of these episodes, two involving Acinetobacter baumannii and one involving Serratia marcescens, correlated with those identified by RAPD-ALF analysis. It was concluded that analysis of antibiogram data alone is an unreliable method for assessing bacterial cross-transmission, unless the organism involved has a particularly stable or unusual resistance pattern. In contrast, the technique of RAPD-ALF analysis may provide a rapid and simple technique for obtaining an insight into the population dynamics of gram-negative bacteria in adult ICUs.

Molecular and antibiogram relationships of Acinetobacter isolates from two contrasting hospitals in the United Kingdom and South Africa.

The aim of this study was to compare the molecular relationships and antibiograms of nosocomial isolates of Acinetobacter spp. from two acute-care hospitals in Nottingham, UK, and Soweto, South Africa, with different hospital infection control problems and procedures. In contrast to Nottingham, where randomly amplified polymorphic DNA fingerprinting demonstrated that a single multiresistant strain of Acinetobacter baumannii has predominated in the hospital intensive care unit over an 11-year period, the Soweto isolates formed a heterogeneous group of unrelated molecular clusters of different antibiograms, with numerous different strains of Acinetobacter baumannii, Acinetobacter sp. 3 and Acinetobacter sp. 13TU apparently being endemic throughout the Soweto hospital. The contrasting results illustrate the need to maintain exemplary infection control procedures in hospitals where high standards have been achieved and warn of what might result if such measures are diminished.

Development and evaluation of a PCR-based immunoassay for the rapid detection of methicillin-resistant Staphylococcus aureus.

A multiplex polymerase chain reaction (PCR), involving detection of the mecA and femB genes, was combined with a novel immunoassay system capable of detecting specific PCR products. The resulting PCR-immunoassay was evaluated in comparison with conventional microbiological techniques used in the routine diagnostic laboratory for the rapid identification of methicillin-resistant Staphylococcus aureus (MRSA), either in pure culture or in overnight broth cultures obtained following enrichment of patient screening swabs. Among the 480 purified isolates of staphylococci and 246 enrichment broths examined, only one 'false-negative' result was obtained by PCR, compared with 18 'false-negative' results obtained by conventional methodology and demonstrated by further conventional examination. Five demonstrable 'false-positive' results were obtained by conventional methodology, compared with a possible 10 by the PCR-immunoassay, although it was not certain that these 10 PCR results were true 'false positives' as, by definition, MRSA could not be isolated by conventional methodology. The results indicated that the routine diagnostic laboratory was encountering difficulties in identifying MRSA correctly, and that the conventional microbiological techniques lacked sensitivity. Overall, the PCR technique was more accurate and sensitive than conventional methodology in detecting MRSA, and results were available within 24 h of screening swabs arriving in the laboratory, compared with a minimum of 48-72 h by conventional techniques. The immunoassay system added to the usefulness of the method by allowing the detection of specific PCR products within 5 min of completing the PCR, without the normal additional step of agarose gel electrophoresis.

Surveillance of an adult intensive care unit for long-term persistence of a multi-resistant strain of Acinetobacter baumannii.

Sporadic infections with Acinetobacter spp., punctuated with prolonged outbreaks of infection involving larger numbers of patients and a particular epidemic strain of Acinetobacter baumannii, have occurred in the adult intensive care unit (ICU) of Nottingham University Hospital since 1985. The aim of this study was to screen patients admitted to the ICU for three or more days during a non-outbreak period in 1994-1995 and to use DNA fingerprinting techniques to compare any isolates of Acinetobacter spp. with isolates obtained from the same ICU during the previous ten years. In the present study, almost 20% of the ICU patients screened during 1994-1995 became colonized with Acinetobacter spp. The commonest species isolated from patients was Acinetobacter baumannii; five different strains were identified by random amplified polymorphic DNA fingerprinting, including the epidemic strain responsible for outbreaks of infection in 1985-1986 and 1992-1993. Environmental sampling yielded Acinetobacter spp. from one or more samples on four occasions; Acinetobacter radioresistens was the commonest species isolated, and Acinetobacter baumannii (not the epidemic strain) was isolated on only one occasion from the environment. The long-term persistence of a potentially epidemic strain in the ICU, even during a non-outbreak period, indicates a need for continued vigilance. Consequently, periodic patient and environmental surveillance, combined with typing of isolates, is recommended for ICUs where significant outbreaks of Acinetobacter infection have occurred in the past.

A novel interaction between inhibitory melatonin receptors and protein kinase C-dependent signal transduction in ovine pars tuberalis cells.

This study revealed an important and unexpected finding: namely, that inhibitory melatonin receptors can inhibit a phorbol 12,13 myristate acetate (PMA)-induced, protein kinase C (PKC)-dependent increase in c-fos messenger RNA expression in ovine pars tuberalis (PT) cells. PMA induces dose-dependent stimulation of c-fos expression that is attenuated by melatonin in a dose-dependent and pertussis toxin-sensitive manner. The effect of 100 nM PMA is blocked by Ro31-8220 (1 microM), yet is not mimicked by 4alpha-PMA (100 nM). PMA (100 nM) induces PKC activity in PT cells (P < 0.05) within 5 min, but melatonin has no effect on this response. PMA (100 nM) stimulates both phospholipase D and mitogen-activated protein kinase (MAPK) (p42/44) activities in PT cells, but melatonin has no effect on these responses. The results indicate that neither of these second-messenger activities contribute to the melatonin-sensitive pathway of c-fos activation. The MEK (MAPK kinase) inhibitor, PD98059 (50 microM), does not block the induction of c-fos by PMA, although at the same dose it inhibits PMA-mediated activation of p42/44 MAPK by 50-70%, and activation by forskolin or insulin-like growth factor-I by 100%. These data suggest that p42/44 MAPK may not be the primary mediator of PKC-dependent c-fos induction. In contrast to the effect of melatonin on PMA-mediated c-fos induction in PT cells, in L cells stably transfected with the sheep Mel1 alphabeta receptor, melatonin potentiates the c-fos response in a pertussis toxin-sensitive manner. These data indicate the tissue-specific nature of melatonin receptor signaling, and reveal that a pertussis toxin-sensitive pathway can block PKC-mediated c-fos induction in PT cells.

The ovine pars tuberalis secretes a factor(s) that regulates gene expression in both lactotropic and nonlactotropic pituitary cells.

The purpose of this study was to determine whether the cells of the ovine pars tuberalis (PT) secrete a factor(s) that can influence the activity of cells in the pars distalis (PD). By Northern blotting of total RNA isolated from PD cells that had been stimulated in the presence of cycloheximide (10 micrograms/ml), PT cell-conditioned medium was shown to induce a significant increase in the expression of the early response gene, c-fos, above both PD cell-conditioned and nonconditioned medium control levels (P < 0.05). Although forskolin (5 microM) induced a weak increase in c-fos expression in PD cells, the effect of PT medium conditioned in the presence of forskolin enhanced this expression more than additively (P < 0.05); furthermore, this effect was reversed by melatonin. These results are consistent with the release of a factor(s) from the PT, which for simplicity we have called tuberalin. This factor was released from PT cells in a time-dependent and cycloheximide-sensitive manner and was resistant to heating at 100 C for 10 min. Tuberalin activity could be size-fractionated using molecular size cut-off filters to produce activity in both the 1- to 10-kDa and more than 10-kDa size ranges. The activities in both of these fractions were sensitive to trypsin degradation and, therefore, appeared to be peptidergic. However, it was not clarified whether the biological activities were due to one or two components. Tuberalin also induced c-fos expression in other cell types, including GH3 and NIH3T3 cells. Dual labelling of PD cells by in situ hybridization using riboprobes for c-fos and PRL demonstrated that both the less than and more than 10-kDa fractions of tuberalin activated c-fos expression in some, but not all, lactotrophs in PD cell cultures, suggesting that a primary function of the PT is to regulate the activity of lactotrophs. This was supported further by enhanced secretion of PRL from PD cells in the presence of either PT-conditioned medium or PT cells in coculture. In addition, PT-conditioned medium was found to increase c-fos in a second cell type, which did not hybridize positively for PRL, indicating the existence of other endocrine interactions between the PT and PD.

Comparative in-vitro activity of cefaclor against urinary tract isolates of Escherichia coli.

Insusceptibility levels of cefaclor and other commonly prescribed antibiotics were determined for 489 consecutive hospital and community-associated urinary tract isolates of Escherichia coli from the Nottingham area of the UK. Significant resistance (MIC of > or = 8 mg/L) to cefaclor was found to be uncommon in the UK, with insusceptibility percentages as low as 1.5% and 1.4% amongst hospital and community isolates, respectively. When compared with other antimicrobials used commonly for treating urinary tract infection, only ciprofloxacin showed greater activity, though cefaclor showed significantly greater in-vitro activity than cephalexin, ampicillin and trimethoprim. Only seven isolates were insusceptible to cefaclor at a concentration of 8 mg/L. Each of these isolates produced a beta-lactamase, but it is unlikely that beta-lactamase production was the sole reason for insusceptibility since these isolates were also insusceptible to co-amoxiclav. Cefaclor compared extremely well with co-amoxiclav against ampicillin-insusceptible isolates, with none of the pharmacokinetic difficulties and considerations associated with the use of beta-lactam:beta-lactamase inhibitor combinations. Cefaclor appears to be a useful cost-effective alternative to current therapeutic options, particularly for long-term low-dose treatment of recurrent urinary tract infection in those geographical areas where the likelihood of resistance to other possible agents is becoming unacceptably high.

Comparison of rapid automated laser fluorescence analysis of DNA fingerprints with four other computer-assisted approaches for studying relationships between Acinetobacter baumannii isolates.

The relationships between isolates suggested by a novel DNA typing method (RAPD-ALFA) that combines randomly amplified polymorphic DNA with automated on-line laser fluorescence analysis of DNA fragments were compared with those suggested by four other computer-assisted typing strategies (biotyping, antibiogram typing, pulsed-field gel analysis of chromosomal fingerprints and arbitrarily-primed DNA amplification with three different primers) for 25 isolates of Acinetobacter baumannii obtained from 12 different hospitals in four countries over a period of 12 years. The results obtained by cluster analysis with two different software packages confirmed that the relationships suggested by RAPD-ALFA were robust and essentially similar to those suggested by the other more laborious computer-assisted typing methods. The technique of RAPD-ALFA appears to offer the possibility of routine on-line molecular identification and typing of isolates from particular hospital wards or units (e.g., intensive care units), and could, therefore, play a key role in the early recognition and prevention of outbreaks of infection.

Stimulation of rat spleen cells by staphylococcal enterotoxins.

There is much interest in staphylococcal enterotoxins as T cell mitogens in humans, mice and rabbits. Rat spleen cells were shown to proliferate in response to staphylococcal enterotoxins A and B and toxic shock syndrome toxin-1 at concentrations (5 to 500 ng ml-1) which also stimulate mouse spleen cells. The proliferative response to all these enterotoxins was inhibited by cyclosporin A, indicating the response to be predominantly that of T cells. These results indicate that the rat provides another convenient model for the analysis of T cell responses to enterotoxins.

Development and design of a novel in vivo chamber implant for the analysis of microbial virulence and assessment of antimicrobial therapy.

An accurate reflection of the pathogenicity of microorganisms and the therapeutic effects of antimicrobial agents on their growth necessitates testing within an in vivo environment. We have developed a novel diffusion chamber, incorporating two 0.22 microns membrane filters, for the growth of in vivo organisms. The chamber, which is implanted intraperitoneally into the rat, has an external sampling portal. This portal allows multiple and sequential sampling of the microbial inoculum without killing the rat, thus significantly reducing the total number of animals used in such studies. In addition, the chamber is superior to other reported implants since it is well tolerated, reusable, easily constructed and can be used within two days of implantation. Staphylococcus epidermidis and a toxic shock syndrome toxin-1 (TSST-1) producing strain of S. aureus have been successfully grown within in vivo chambers, with 10(8)-10(9) organisms per millilitre being recovered within 48 h. Scanning electron microscopy revealed clusters of staphylococci and fibrous material adhering to the inner surface of the filters, with numerous phagocytic cells attached to the outer side. Western immunoblotting indicated that higher levels of TSST-1 were produced by S. aureus grown in vivo as opposed to cells grown in vitro.

4-[2-(Di-n-propylamino)ethyl]-2(3H)-indolone: a prejunctional dopamine receptor agonist.

4-[2-(Di-n-propylamino)ethyl]-2(3H)-indolone (1c) (SK&F 101468) is a potent and selective prejunctional dopamine receptor agonist. It caused a dose-related inhibition of the constrictor response to electrical stimulation in the isolated perfused rabbit ear artery (EC50 = 100 nM), and this response was antagonized by (S)-sulpiride (KB = 7 nM). Compound 1c did not stimulate or block dopamine-sensitive adenylate cyclase and did not produce stimulation of the central nervous system in rats. It was prepared from (2-methyl-3-nitrophenyl)acetic acid in a multistep sequence based on the Reissert indole synthesis.

Comparative rating of consultation performance: a preliminary study and proposal for collaborative research.

Factors governing the appropriateness, reliability and validity of rating scales in the measurement of professional performance are reviewed. The origin and preliminary testing among undergraduated and general practitioners of a brief consultation rating schedule is described.Statistical criteria are proposed for the analysis of ratings, by groups, in the comparison of consultation performance. Using these criteria the capacity of the 10 rating schedule items to discriminate between two contrasting consultations was examined. Each of the items was used at some time by students or doctors to express significant preference for the same consultation; and on this basis all the items are considered to merit inclusion. One item showed highly significant intra- and inter-observer reliability.The schedule is reproduced in full, together with a data-collection document and significance chart, with the aim of encouraging groups of doctors to test the validity of the items in the comparison of other pairs of consultations. It is proposed that future versions of the schedule should reflect the experience of such groups in testing existing items and in defining additional items which satisfy the proposed criteria.

Trimethoprim resistance plasmids in Escherichia coli isolated from cases of diarrhoea in cattle, pigs and sheep.

A total of 1572 isolates of Escherichia coli obtained from the faeces of young farm animals with diarrhoea over the period 1980-1983 were screened for resistance to trimethoprim (Tp). Resistance to Tp was detected in 263/954 (28%) of bovine isolates, 59/441 (13%) of porcine isolates and 15/177 (9%) of ovine isolates. Seventy-five resistant isolates from separate outbreaks of infection on farms within a 25 mile radius of Nottingham were examined in detail. Sixty-eight (91%) of the 75 isolates were resistant to greater than 1024 mg Tp/l and 34 (50%) of these 'highly resistant' isolates (45% of total resistant isolates) transferred their Tp resistance to E. coli K12. A further 13 (17%) isolates were demonstrated to carry non-self-transferable plasmids which were capable of being mobilized to E. coli K12 by the broad host range plasmid RP4. Thirty-one self-transferable Tp R plasmids were divided between the following incompatibility groups: IncB (14 plasmids), IncFII (4 plasmids), IncH2 (1 plasmid), IncI alpha (10 plasmids), IncI delta (1 plasmid) and IncP (1 plasmid). In terms of antibiotic resistance patterns and incompatibility properties, many of these plasmids closely resembled those isolated from human patients in the same area, suggesting that there may be a common pool of Tp R plasmids.