Mobile technology in clinical teaching.

Technology is having a profound effect on education in the 21st century and nurse educators are being challenged to integrate technological innovation to assist students in their learning. This paper reports a study on the introduction of smart mobile technology to support student learning in the clinical environment. In a climate of collaborative inquiry, clinical lecturers and two researchers from the same department carried out a project in three phases: formation, implementation and analysis. Following the formation phase, six clinical lecturers adopted iPads to support their clinical teaching (implementation phase). At this time they also kept reflective journals. In the analysis phase a thematic analysis of the data from the journals and from a focus group found both enabling and constraining factors influenced the use of iPads by clinical lecturers. The themes categorised as enablers were: resources and technology; and, management and technology support. Those identified as barriers or constraining factors were: clinical staff engagement; and lecturer experience with technology. Student engagement and learning, and connectivity were both enabling and constraining factors. This paper concludes that the use of a mobile device such as an iPad can enhance teaching in clinical settings but that in order for such devices to be successfully integrated into clinical teaching consideration needs to be given to professional development needs, adequate resourcing and technology support.

Assessing the efficacy of three dentifrices in the treatment of dentinal hypersensitivity.

A multicenter clinical trial conducted by the authors compared the desensitizing efficacy of a new 5 percent potassium nitrate: 0.243 percent sodium fluoride dentifrice along with two clinically proven, commercially available desensitizing dentifrices to a placebo dentifrice. Sensitivity to cold air and tactile stimulation, along with patients' subjective assessments, were evaluated to assess the dentinal desensitizing efficacy of the test dentifrices. Results demonstrated that after four weeks, participants who used the new dentifrice formulation experienced significant decreases in dentinal sensitivity compared to the placebo group for all measured indexes.

A ribozyme which discriminates in vitro between PML/RAR alpha, the t(15;17)-associated fusion RNA of acute promyelocytic leukemia, and PML and RAR alpha, the transcripts from the nonrearranged alleles.

Acute promyelocytic leukemia (FAB M3) is distinguished by the presence of the t(15;17) and clinical response to all-trans retinoic acid (RA) treatment. Acute promyelocytic leukemia is associated with a chromosomal translocation which results in the fusion of genes encoding a putative transcription factor (PML) and the retinoic acid receptor alpha (RAR alpha). It is suggested that the PML/RAR alpha fusion protein functions as an inhibitor of myeloid differentiation. The potential use of ribozymes as therapeutic agents has been investigated in the present study. Hammerhead ribozymes, which by hybridizing to both PML and RAR alpha sequences discriminate between the fusion transcript and the normal transcripts from the nonrearranged alleles, were designed and synthesized. Two hammerhead cleavage sites were targeted: site 1, an AUU located 4 nucleotides 3' to the fusion junction; and site 2, a UUC located 26 nucleotides 3' to the junction. Both sites are located in the RAR alpha portion of the fusion transcript. Using a full-length PML/RAR alpha RNA or an RNA corresponding to 788 nucleotides of the PML/RAR alpha mRNA and a full-length RAR alpha RNA or an RNA corresponding to 960 nucleotides of the RAR alpha mRNA as model substrates, the catalytic behavior of several ribozymes was studied. A modified hammerhead directed against site 2 displayed the highest degree of selectivity for PML/RAR alpha. It is hypothesized that ribozyme-mediated inactivation of PML/RAR alpha provides a new approach to study the role of PML/RAR alpha in the deregulated growth and RA response of acute promyelocytic leukemia.

Integrating nursing research and practice: Part I--A journey into the unknown.

We must all face the challenge of weaving together the art and science of nursing, of integrating nursing research and practice. In a two part article Northland nurses tell about their experiences with research and the study they carried out. Part one describes their beginning experiences with integrating research and practice.

Lysozyme-mediated de-chaining of Streptococcus mutans and its antibacterial significance in an acidic environment.

The ability of physiological amounts of lysozyme to de-chain two serotype c strains of Streptococcus mutans was determined. Both human and hen lysozymes were equally effective in chain breakage of S. mutans DPR and S. mutans DJR. De-chaining did not affect growth of cultures, but resulted in finely dispersed suspensions, at stationary phase, which were visibly different from untreated cultures. Less than 50 micrograms lysozyme per ml culture medium reduced chain length to virtually all diplococci and single cells, and this chain disruption increased total viable cell count. De-chaining required an active enzyme indicating that a degree of hydrolysis of the peptidoglycan occurred at the septae of the streptococci. De-chained S. mutans did not survive as well as streptococci of normal chain length when incubated under acidic conditions (pH 5.5), but gross cellular lysis was not apparent. The reduced aciduric property of the disrupted chains may have been due to a participation of autolysins or to a lethal triggered by the lysozyme-damaged peptidoglycan. De-chaining may be a mechanism by which lysozyme could regulate the levels of S. mutans in acidogenic plaque samples.

Fungistatic and fungicidal activity of human parotid salivary histidine-rich polypeptides on Candida albicans.

Human parotid saliva histidine-rich polypeptides exerted antifungal activity against Candida albicans at concentrations similar to the known antifungal activity of the imidazole antibiotics. Inhibition of both growth and viability could be demonstrated by optical density monitoring and plating assays. Inhibition of growth was observed to be greatest when the histidine-rich polypeptides were added to the inoculum before addition to the growth media. However, complete inhibition by these polypeptides was still noted during active growth at turbidities of C. albicans corresponding to 10(6) CFU/ml. At higher cell densities, growth was delayed but not halted under the experimental conditions investigated. Candidacidal activity was observed with both growing and nongrowing cells. With respect to the latter, reaction of cells in buffer with the histidine-rich polypeptides for a period of 30 min resulted in killing of greater than 90% of two different strains of C. albicans, whereas a third strain was found to be less susceptible. Moreover, the kinetics of loss of cell viability correlated with the loss of potassium from the cells. In addition to the histidine-rich polypeptides, hen egg white lysozyme, poly-L-lysine, and poly-L-histidine affected C. albicans. Both of the polyamino acids completely inhibited the growth of the yeast whereas lysozyme was not as potent. Where delays in growth were observed for all of these agents, including the histidine-rich polypeptides, turbidities reached those of untreated controls after a 24-h period. Enhanced effects were noted if C. albicans was preincubated with these agents in 0.025 2-(N-morpholino)-ethanesulfonic acid buffer, pH 5.2, before growth in the yeast synthetic medium.

Isolation of milligram quantities of a group of histidine-rich polypeptides from human parotid saliva.

Freshly collected parotid saliva collected from human donors were shown by polyacrylamide gel electrophoresis to continuously secrete a group of low-molecular-weight cationic polypeptides. Up to 14 bands could be identified by Coomassie blue staining, and all bands migrated more rapidly than purified human leukemic lysozyme in cationic polyacrylamide gel electrophoresis. These peptides could be isolated as a group relatively free of other salivary components and recovered in high yields from concentrated parotid saliva by Sephadex G-25 chromatography. In sodium dodecyl sulfate gel electrophoresis, the histidine-rich polypeptide bands appeared as just two bands migrating at the tracking dye and ahead of insulin chain B. Amino acid analysis of the mixture revealed an average content of at least 48% cationic residues, of which half were histidine. When stained bands were eluted from electrophoretic gels, hydrolyzed, and subjected to amino acid analyses, they were found to be enriched in histidine. There was also a correlation of the electrophoretic mobility with the content of basic amino acids. Sephadex G-25 chromatography is a convenient, simple method for preparing milligram quantities of the histidine-rich polypeptides for chemical and biochemical studies.

Growth-inhibitory and bactericidal effects of human parotid salivary histidine-rich polypeptides on Streptococcus mutans.

Growth inhibition and cell viability assays demonstrate that the histidine-rich polypeptides isolated from human parotid saliva are bacteriostatic and bactericidal for strains of Streptococcus mutans belonging to the serotype b and c classifications. Both inhibition of growth and cell division are enhanced by preincubation of bacteria with these polypeptides in low-ionic-strength buffers of acidic and neutral pH before dilution into enriched growth media. With prior exposure at pH 6.8, inhibition by these polypeptides of the serotype c strains, S. mutans GS5 and SB, as well as the serotype b strain, S. mutans BHT, is reversible over time under the experimental conditions selected. With similar exposure at pH 5.2, however, irreversible damage is manifested by complete inhibition of both growth and cell viability. At concentrations of 250 micrograms of the mixture of histidine-rich polypeptides per 5 X 10(5) bacterial cells per ml in the acidic preincubation buffer, bacterial lethality is maintained for a period of 48 h in the enriched growth media. At a 50-micrograms/ml concentration of these salivary agents, approximately 80% killing of S. mutans SB is noted after a 24-h incubation; however, surviving bacteria multiply and reach turbidities of untreated control cells when examined at the 48-h growth point. Similarly, hen egg white lysozyme is also found to be bactericidal for these microorganisms when preincubation is carried out under acidic conditions. However, in contrast to the histidine-rich polypeptides, lysozyme under these experimental conditions does not inhibit growth of S. mutans SB at neutral pH, although it does inhibit growth of both S. mutans BHT and S. mutans GS5 at this pH. Preexposure of S. mutans SB to the peptides in buffer at ionic strengths of 0.025 to 0.125, followed by either viability assays under nongrowing conditions or growth inhibition studies, suggests that there is very little effect of ionic strength on the antibacterial function of these peptides. In contrast to the inhibition of viability noted under growing conditions, lower concentrations of the histidine-rich polypeptides were required to elicit immediate cell death under nongrowing conditions.

Development of an enzyme-linked immunosorbent assay for determination of lysozyme in human parotid and submandibular-sublingual salivas.

The specificity of lysozyme determinations in human parotid and submandibular-sublingual salivas of two subjects was assessed by comparison of lysozyme concentrations in native acidified salivas with purified enzyme obtained by immunoadsorbent fractionation of the salivas. Lysozyme concentrations were measured by the turbidimetric catalytic method and by a newly developed enzyme-linked immunosorbent assay (ELISA). The validity of the assays was established by comparing assay results with enzyme concentration values determined from optical density-extinction coefficient calculations of the purified lysozyme peak. Values for purified enzyme were found to be similar, irrespective of the assay used to determine lysozyme concentrations, and were in agreement with extinction coefficient calculations. Based on the ELISA technique, recoveries of lysozyme from both parotid and submandibular-sublingual salivas were greater than 75 and 90%, respectively. Similar recoveries were noted for parotid saliva when determinations were based on the turbidimetric assay. However, the ELISA and turbidimetric assays differed with respect to lysozyme levels in submandibular-sublingual saliva because of the apparent presence of an enhancement factor which gave rise to higher lysozyme values in the catalytic assay and therefore resulted in low recoveries of purified enzyme. This catalytic enhancement factor was present in the nonadsorbed fraction of both subjects, as higher lysozyme activities were noted when nonadsorbed fractions were added to affinity-purified lysozymes. Lysozyme levels were also determined in the parotid and submandibular-sublingual salivas of caries-resistant and -susceptible adults. In general, levels of lysozyme in parotid saliva were lower in comparison to submandibular -sublingual saliva; however, significant differences in enzyme concentration were not evident between the caries-resistant and caries-susceptible subjects.

Lytic sensitivity of Actinobacillus actinomycetemcomitans Y4 to lysozyme.

The ability of both human and hen egg white lysozymes to lyse Actinobacillus actinomycetemcomitans Y4 was investigated. Lysis was followed optically at 540 nm by measuring the percent reduction in turbidity of freshly harvested log-phase cells suspended in Tris-maleate buffers within a wide range of pH (5.2 to 8.5) and molarity (0.01 to 0.2 M) and containing various amounts of enzyme and EDTA. In several instances, treated microorganisms were subsequently examined in thin sections by electron microscopy. Reductions in turbidity and clearing of suspensions occurred with small amounts of lysozyme (less than 1 microgram) under relatively alkaline conditions and at low ionic strength and in the presence of small amounts of EDTA (greater than 0.01 mM). Under the most alkaline conditions, EDTA alone effected turbidity reductions similar to those observed in the presence of lysozyme, which suggested that EDTA not only increased outer membrane permeability but also caused cell lysis. Ultrastructural analysis did not always correspond to turbidimetric observations. Cell lysis was virtually complete in suspensions containing both lysozyme and EDTA. However, in contrast to turbidimetric findings, a significant percentage of cells (greater than 25%) was lysed in the presence of lysozyme alone. Furthermore, significant damage occurred in the presence of EDTA alone. Spheroplast-like cell ghosts were present which surrounded condensed cytoplasm or relatively clear spaces. These findings further support the concept of the requirement for electron microscopy to assess lytic damage in addition to turbidimetric and biochemical methods. Our results are the first to demonstrate the remarkable sensitivity of A. actinomycetemcomitans Y4 to lysozyme and to show that EDTA not only affects outer membrane permeability but effects cell lysis, possibly through activation of autolytic enzymes at the cytoplasmic membrane. The exquisite sensitivity of A. actinomycetemcomitans Y4 to lysis could be an important mechanism by which lysozyme participates in the regulation of this suspected periodontal pathogen.

Plasma prolactin levels and body fluid deficits in the rat: causal interactions and control of water intake.

A study was made of the responses of chronically hyperprolactinaemic rats to selected dipsogenic stimuli. Measurements were also made of the correlation between the state of hydration of the animal and the plasma prolactin levels. After 24 h water deprivation. S.C. isoprenaline (10 micrograms/kg body wt.) or I.P. injection (5 ml/kg body wt.) of a hypertonic solution (50% w/w) of polyethylene glycol (mol. wt 20000) there was no difference between the hyperprolactinaemic and control rats with respect to the total water intake, the time course of drinking or the urine output. After I.V. injection of 2 M-NaCl (5 ml/kg body wt.) there was no difference between the hyperprolactinaemic and control rats with respect to the total water intake or urine output. However, the hyperprolactinaemic rats drank more slowly than the controls. When angiotensin II was infused I.V. at a rate of 0.2 micrograms/min, the water intake was greater and the threshold to drinking lower in the hyperprolactinaemic than control rats. After 24 h water deprivation, plasma prolactin levels rose significantly in both the control and hyperprolactinaemic rats. When the rats were injected I.V. with hypertonic saline (5 ml, 2 M-NaCl/kg body wt.) and denied access to water, plasma prolactin levels had not changed 1 h later in either the control or hyperprolactinaemic animals. It is concluded that there is no interaction either between the plasma osmolality and prolactin secretion or between the plasma prolactin levels and the amount of water drunk in response to intracellular fluid deficits. However, prolactin secretion is stimulated by the combined intra- and extracellular deficits resulting from water deprivation and there is a clearly demonstrated interaction between prolactin and the extracellularly mediated stimulus of angiotensin II.

Bacteriolysis of Streptococcus mutans GS5 by lysozyme, proteases, and sodium thiocyanate.

Streptococcus mutans GS5 was grown in a synthetic medium containing radioactive thymidine to monitor cell lysis by assay of the release of DNA. Bacteriolysis was achieved by sequential treatment of the cells with either hen egg white lysozyme and sodium thiocyanate or a combination of hen egg white lysozyme and a proteolytic enzyme followed by addition of the thiocyanate. In the absence of sodium thiocyanate, a small percentage of the total macromolecular thymidine was released in control reaction mixtures during incubation. This amount of released DNA more than doubled in trypsin-treated cells, but the inclusion of lysozyme in reaction mixtures prevented assay of the DNA. Lysis was found to be optimal in the late log phase of growth and was dependent on the concentrations of both lysozyme and protease. Concentrations of trypsin or chymotrypsin as low as 0.01 microgram/ml were found to be effective in enhancing the lytic process. The addition of protease to lysozyme-inorganic salt reaction mixtures altered both the pH and ionic strength profiles of cell lysis. At pHs of 5.5 or lower, both the lysozyme-NaSCN and the lysozyme-trypsin-NaSCN systems were inactive in mediating lysis. The loss of insoluble cell wall peptidoglycan by lysozyme treatment was pH independent and did not appear to be affected by the addition of protease. Either diluted whole saliva or neutrophil extracts could replace trypsin to enhance cell lysis further.

Quantitative recovery, selective removal and one-step purification of human parotid and leukemic lysozymes by immunoadsorption.

Immunoadsorption affinity chromatography was used to isolate and purify human lysozyme. The immunoadsorbent was prepared by coupling sheep anti-(human leukemic lysozyme) IgG to epoxy-activated Sepharose 6B. Lyophilized parotid saliva (21) was resuspended in distilled water (325 ml, 50 mg/ml, w/v) and applied to a column which had a capacity to bind 4.25 mg human enzyme. Non-adsorbed material did not contain lysozyme, as determined by enzymatic and immunological analyses. All lysozyme activity present in the applied sample (1.97 mg) bound to and was desorbed from the column by elution with 0.2 M sodium acetate HCl buffer, pH 1.8. The isolated material was homogeneous as determined by cationic and sodium dodecyl sulfate/polyacrylamide gel electrophoresis, ultracentrifugation, amino acid and amino-terminal analyses, and immunoelectrophoretic analysis. The one-step purification procedure yielded a 1370-fold increase in specific activity. Human lysozyme was also selectively purified by this method from an ammonium sulfate precipitate of the urine of a patient with chronic monocytic leukemia. Amino acid and polyacrylamide gel electrophoretic analyses indicated that the purified enzyme was identical to human lysozyme isolated from leukemic urine by classical biochemical techniques.

Lysozyme binding by a polyglycerol phosphate polymer of the oral bacterium Streptococcus mutans BHT.

The antibacterial properties of lysozyme for Streptococcus mutans BHT may be a function of its binding to cell components other than to peptidoglycan. Inhibitors of muramidase activity, including histamine and N-acetyl-D-glucosamine, only partially blocked the bacteriostatic effects on this strain. Greater than 20 mM histamine alone inhibited growth suggesting a bacteriostatic potential. An autoclaved saline extract was then prepared from stationary phase cultures in a chemically-defined medium. As little as 31.25 micrograms of the extract significantly blocked the effect of 50 micrograms lysozyme and complete enzyme inhibition was achieved with 62.5 micrograms. The extract was fractionated and location of potential binding components determined by a precipitin method consisting of diffusing the samples into 1.2 per cent agarose containing lysozyme. Binding components eluted in the first peak of a Sephacryl S-300 column, bound to DEAE-cellulose, but desorbed with gradient elution (0.1-1.0 M tris-HCl buffer, pH 8.0). The eluted material was then applied to an affinity column containing purified lysozyme coupled to epoxy-activated Sepharose 6B. Non-absorbed anionic material precipitated only with protamine. Lysozyme-binding fractions eluted in a sharp peak with 1.0 M tris-HCl buffer (pH 8.0), did not bind wheat-germ agglutinin, contained less than 50 micrograms protein, 95 micrograms sugar, 66.7 micrograms phosphorus, less than 0.25 mequiv lipid and no detectable nucleic acids. The peak material reacted with antiserum directed against polyglycerol phosphate, indicating that it contained acylated or, possibly, deacylated lipoteichoic acid. The findings suggest that the antibacterial properties of lysozyme for Strep. mutans BHT may, in part, be modified (or possibly regulated) by binding to molecules such as lipoteichoic acid.

Daily water and electrolyte balance in chronically hyperprolactinaemic rats.

1. Chronic hyperprolactinaemia was induced in eight male rats by implantation of anterior pituitary glands from inbred female rats. The ten control rats underwent sham operations. 2. The spontaneous 24 hr water intake of hyperprolactinaemic rats was greater than that of the controls. Intake was increased only at night. 3. The increased water intake was independent of food intake and was not secondary to increased salt intake. 4. When prolactin secretion was inhibited with bromocriptine, there was a parallel fall in water intake and plasma prolactin levels in the experimental animals to the levels observed in the controls. 5. Urine volume was greater and urine osmolality lower in the hyperprolactinaemic rats than in the controls during both day and night. 6. In the evening, plasma osmolality was lower in the hyperprolactinaemic animals than in the controls. No such difference was observed in the morning. 7. Blood volume was greater in the hyperprolactinaemic rats than in the controls, but the haematocrits were the same. 8. These findings indicate prolactin may have a primary dipsogenic activity.

Selective antibacterial properties of lysozyme for oral microorganisms.

The antibacterial properties of lysozyme were investigated with oral microorganisms representing the seven serotypes (a through g) of Streptococcus mutans, Veillonella alcalescens, and the virulent (V) and avirulent (AV) strains of Actinomyces viscosus T14. Growth of bacteria in defined medium was monitored spectrophotometrically after the addition of various amounts (25 mug to 5 mg/ml) of enzyme. No growth inhibition of V. alcalescens was observed. Inhibition of A. viscosus T14(V) and A. viscosus T14(AV) occurred with 160 mug of lysozyme per ml. Of the S. mutans cultures tested, the serotype a and b strains were inhibited with as little as 25 mug of enzyme per ml, whereas e and f strains were most resistant to the bacteriostatic activity of lysozyme. The presence of dl-threonine or sucrose in growth medium did not significantly affect the results. A lysoplate assay was developed to rapidly survey the bacterial cultures for their susceptibility to the lytic ability of the enzyme. Lysis, as a measure of a zone of clearing in agarose plates, occurred for all microorganisms in the presence of lysozyme after the subsequent addition of NaCl or detergent. The bactericidal activity of lysozyme was determined on S. mutans BHT and S. mutans LM-7 by the pour plate technique. Preincubation of S. mutans LM-7 with as much as 1 mg of enzyme for 90 min did not affect viability or growth, whereas preincubation of S. mutans BHT with 1 mg of lysozyme resulted in no recoverable colony-forming units. An antigen containing extract of S. mutans LM-7 blocked the growth inhibitory property of lysozyme. Human lysozyme was a more effective antibacterial factor than hen egg white lysozyme. Total growth inhibition of S. mutans BHT was effected with 40 mug of human enzyme, and as little as 10 mug of human enzyme inhibited growth for greater than 20 h. The data presented indicate that different mechanisms may be responsible for the bacteriostatic, lytic, and bactericidal properties of the enzyme and that lysozyme is a selective but effective antibacterial factor for oral microorganisms.