DNA-based immunization by in vivo transfection of dendritic cells.

Delivery of antigen in a manner that induces effective, antigen-specific immunity is a critical challenge in vaccine design. Optimal antigen presentation is mediated by professional antigen-presenting cells (APCs) capable of taking up, processing and presenting antigen to T cells in the context of costimulatory signals required for T-cell activation. Developing immunization strategies to optimize antigen presentation by dendritic cells, the most potent APCs, is a rational approach to vaccine design. Here we show that cutaneous genetic immunization with naked DNA results in potent, antigen-specific, cytotoxic T lymphocyte-mediated protective tumor immunity. This method of immunization results in the transfection of skin-derived dendritic cells, which localize in the draining lymph nodes. These observations provide a basis for further development of DNA-based vaccines and demonstrate the feasibility of genetically engineering dendritic cells in vivo.

A probe for in situ, remote, detection of defects in buried plastic natural gas pipelines.

Several techniques are available to determine the integrity of in situ metal pipeline but very little is available in the literature to determine the integrity of plastic pipelines. Since the decade of the 1970s much of the newly installed gas distribution and transmission lines in the United States are fabricated from polyethylene or other plastic. A probe has been developed to determine the in situ integrity of plastic natural gas pipelines that can be installed on a traversing mechanism (pig) to detect abnormalities in the walls of the plastic natural gas pipeline from the interior. This probe has its own internal power source and can be deployed into existing natural gas supply lines. Utilizing the capacitance parameter, the probe inspects the pipe for flaws and records the data internally which can be retrieved later for analysis.

Ability of physiotherapists to undertake evidence-based practice steps: a scoping review.

BACKGROUND: Evidence-based practice (EBP) is promoted to ensure quality of care. However, analysis of the skill of physiotherapists in undertaking the steps of EBP, or the impact of EBP on the work of physiotherapists is limited. OBJECTIVES: To conduct a scoping review into physiotherapists performing the steps of EBP. DATA SOURCE: Literature concerning the skill of physiotherapists in EBP between 1990 and June 2013 was searched using AMED, Academic Search Complete, CINAHL, PubMed, ERIC, PEDRO and EMBASE databases. STUDY SELECTION: Twenty-five studies (six qualitative, one mixed methods and 18 quantitative) were selected. DATA EXTRACTION AND SYNTHESIS: Quantitative and qualitative data were extracted using two appraisal tools to analyse each of the five steps of EBP. RESULTS: Limited evidence exists to show that physiotherapists undertake the full EBP process. Despite formulating clinical questions and acquiring literature-based evidence, the drivers for conducting literature or evidence searches have not been clarified. The critical appraisal step was mainly assessed in the form of recognition of statistical terms. Only examples of guideline usage support the reflective final assessment step. Physiotherapists report using their peers and other trusted sources in preference to literature, primarily due to time but also due to divergence between the literature-based evidence and other evidence that they use and value (tacit knowledge). A positive impact of EBP on patient outcomes is lacking. CONCLUSIONS: Understanding the information needs of physiotherapists may be necessary before adoption of the EBP process. The use of professional networks may offer a better means to identify knowledge gaps and translate acquired knowledge into practice, rather than focusing on individual skills in EBP.

Processing of the Bacillus subtilis thrS leader mRNA is RNase E-dependent in Escherichia coli.

We have recently reported that processing occurs in the untranslated leader region of several members of a family of Gram-positive genes regulated by tRNA-mediated antitermination. We showed that cleavage at this site plays an important role in the induction of Bacillus subtilis thrS gene expression, following threonine starvation, by stabilising the downstream mRNA. Here we show that, when transferred on a plasmid, processing of the B. subtilis thrS leader can occur at the same site in Escherichia coli. Cleavage at this site is dependent on the E. coli endoribonuclease E, both in vivo and in vitro, suggesting that a functional homologue of RNase E is responsible for thrS processing in B. subtilis.

Aminoacyl-tRNA synthetase gene regulation in Bacillus subtilis.

In this review, we summarize progress on the regulation of the aminoacyl-tRNA synthetase genes in Bacillus subtilis. Most of the genes encoding this set of enzymes in B subtilis are members of a large family of Gram-positive genes and operons controlled by a novel antitermination mechanism that uses their cognate uncharged tRNA as the effector. A subset of these genes is, in addition, likely to be controlled at the level of mRNA processing and degradation. We describe the key experiments leading to these conclusions.

Habituation produces frequency-specific plasticity of receptive fields in the auditory cortex.

Associative learning produces conditioned stimulus (CS)-specific plasticity of frequency receptive fields (RFs) in the auditory cortex; responses to the CS frequency are increased, whereas responses to other frequencies are decreased. This study determined the effects of habituation on the RF of neurons in the auditory cortex of the guinea pig (Cavia porcellus). One frequency was presented repeatedly (REP) followed by redetermination of the RF. After REP, 26/36 (72%) RFs exhibited a substantial reduction (70-75%) of response to the repeated frequency, and this was highly specific (bandwidth less than 0.125 octave). This RF plasticity involves an initial decrease in response during REP but does not require attenuated responses at the end of REP. Incubation (i.e., development over time after cessation of REP) and long-term frequency-specific effects are evident. Thus, habituation induces a specific change in the processing of frequency information rather than a general reduction in responsivity.

Neuropeptide modification of chloride secretion in guinea pig distal colon.

This study examined the effects of electrically stimulating submucosal neurons in the guinea pig isolated distal colonic mucosa and determined the effects of several peptides that are present in these neurons. Electrical field stimulation of muscle-stripped segments of guinea pig distal colonic mucosa, set up in Ussing flux chambers, evoked an increase in short-circuit current (Isc), of 371 +/- 31 MicroA.cm-2. The response to electrical stimulation was abolished by tetrodotoxin and significantly reduced by serosal furosemide. Atropine reduced, but did not abolish, the neurally evoked response. Addition of neuropeptide Y and galanin to the serosal bath had no effect on baseline Isc, but both evoked a concentration-dependent decrease in the neurally evoked secretory response. Vasoactive intestinal polypeptide evoked a concentration-dependent increase in basal (unstimulated) Isc that was reduced by furosemide and unaltered by tetrodotoxin. Neuropeptide Y, but not galanin, significantly reduced the secretory responses to vasoactive intestinal polypeptide and bethanechol. Somatostatin 201-995 and human calcitonin gene-related peptide had no effect on basal Isc nor did either alter the neurally evoked response. These results suggest that acetylcholine and non-cholinergic neurotransmitter(s) stimulate chloride secretion in the guinea pig distal colonic mucosa. This neurosecretory response may be modulated by neuropeptide Y and galanin that are found within submucosal neurons.

Classification of the temporal discharge patterns of single auditory neurons in the frog superior olivary nucleus.

Temporal discharge patterns of neurons in the superior olivary nucleus (SON) of the northern leopard frog (Rana pipiens pipiens) were studied by evaluating peri-stimulus time histograms and interspike interval histograms generated from responses to tone bursts at the neuron's characteristic frequency and at 10 dB above neuron's characteristic threshold. Four basic discharge patterns were observed, i.e., primary-like, phasic-burst, phasic, and pauser. Additionally, within each class, different neurons exhibited further subtle differences in temporal discharge patterns and thus subclasses were distinguishable. For most SON neurons, changes in signal level did not affect the discharge pattern. However, there were a few notable exceptions. The firing pattern of 12% of SON neurons changed from one class to another with a change in signal level. Two-thirds of phasic-burst neurons showed increased regularity (i.e., chopping pattern) in their discharges as the signal level was increased. The majority of SON neurons showed monotonic rate-level functions, but one-fourth gave non-monotonic rate-level functions. The presence of a fairly large number of non-monotonic units in the frog SON suggests an additional role of this nucleus in information processing beyond the simple coding of stimulus level. The discharge patterns displayed by SON neurons resembled those seen in the frog cochlear nucleus and in the mammalian superior olivary complex.

Late onset hepatitis and prolonged deterioration in hepatic function associated with nevirapine therapy.

The aetiology of hepatic dysfunction in patients with HIV infection is multifactorial. Re-activation of hepatitis C infection, drug toxicity, and opportunistic infections are all potential causes. Nevirapine (NVP) is a non-nucleoside reverse transcriptase inhibitor used as part of combination antiretroviral therapy for the treatment of HIV infection. It is associated with a significant incidence of hepatotoxicity, usually occurring in the initial month of therapy. We report the case of a 49-year-old man who developed NVP-induced prolonged hepatotoxicity 5 months after commencing antiretroviral therapy.

Rapid production of a panel of blood group A-active oligosaccharides using chemically synthesized di- and tri-saccharide primers and an easily prepared porcine (1-->3)-alpha-N-acetyl-D-galactosaminyltransferase.

A porcine (1-->3)-alpha-N-acetyl-D-galactosaminyltransferase was obtained in a state suitable for preparative-scale (mg-scale) synthesis using simple procedures requiring only three days of effort. The enzyme thus prepared transferred GalNAc efficiently from UDP-GalNAc to six different chemically synthesized di- and tri-saccharide H-active structures to yield blood-group A-active oligosaccharides that were characterized by 1H NMR spectroscopy and mass spectrometry. This work further demonstrates the efficiency and attractiveness of using glycosyltransferases in a combined chemoenzymatic approach for the rapid production of biologically active oligosaccharides.

Does the rose still smell as sweet? Item variability across test forms and revisions.

This article examines item stability when the same item appears in different contexts. The 1st section considers the assumptions in classical test theory and item response theory concerning the relationship between the item and the trait it is presumed to measure. The 2nd section presents contextualist challenges to the measurement theory assumptions about item properties and shows the instability of item characteristics across different testing contexts. The 3rd section describes methods for checking the relationship between items and traits. Classical test methods, item response methods, and structural equation methods for assessing item stability are reviewed. The instability of item characteristics across contexts should caution researchers to assess, and not assume, that items operate the same way on different test versions. Item instability also indicates the need for a more detailed understanding of the psychological processes that occur between item and answer.

Extensive unstable psoriasis--morbidity and medical management.

Over a 5 year period, from January 1988 to January 1993, 23 patients were admitted with extensive unstable psoriasis, 21 of whom had full case notes available. Over the study period these 21 patients required 45 admissions, the mean duration of each admission being 30.54 days. Those patients living more than 30 miles from our unit required almost twice as many admissions as the city group. In 77% of cases, a precipitating factor for the acute flare was identified, and in 53% of these the precipitating factor was iatrogenic related. These patients are prone to develop complications of which infection is the most common. The complexity of medical management was reflected in the number of cross consultations requested. These findings have implications for the optimal delivery of care to this important sub-group of psoriatics.

A retrospective study of myocardial infarction occurring in elderly patients in a coronary care unit.

Myocardial infarction is a major cause of morbidity and mortality particularly in the elderly. Coronary Care Unit monitoring has resulted in the improvement in prognosis in all age groups. In this study, the clinical course and outcome of 204 consecutive patients with acute myocardial infarction aged over 65 years is retrospectively analysed. All patients were managed in a Coronary Care Unit operating a needs related policy. Those under 75 years (120 patients) were compared to those over that age (84 patients). The groups differed in terms of clinical presentation, cardiac enzymatic rise and response to drug therapy. However they had similar complication rates, morbidity and mortality.

Long term effects of smoking cessation program for cardiac patients ... (continuing education credit).

Twenty-eight cardiac rehabilitation patients who were smokers were followed for three years to measure smoking outcomes after an inpatient smoking cessation intervention. Participants were interviewed at three weeks, six months, one, two, and three year intervals to determine their smoking practice. Thirty-six percent (N = 10) of the smokers were not smoking at three years post education. The results suggest that smoking intervention can have a positive impact on long term smoking cessation outcomes.

Round one of the Adelaide and Meath Hospital/Trinity College Colorectal Cancer Screening Programme: programme report and analysis based on established international key performance indices.

BACKGROUND: In Ireland, colorectal cancer (CRC) is the second most frequently diagnosed cancer in men, after prostate cancer, and the second most frequently diagnosed cancer in women, after breast cancer. By 2020, the number of new cases diagnosed annually in Ireland is projected to have increased by 79% in men and 56% in women. Organised screening for CRC is already underway or is in the process of being rolled out in several European countries, either at a regional or national level. The Adelaide and Meath Hospital/ Trinity College Dublin Colorectal Cancer Screening Programme (TTC-CRC-SP) is Ireland's first pilot population based bowel screening programme. METHOD: Based on a biennial test model the pilot aimed to assess the accuracy of FIT and to evaluate the whole programme based on established international key performance indices. RESULTS: To date 9,993 individuals aged 50-74 years have been invited to participate in the TTC-CRC-SP with over 5,000 FIT's analysed. Overall uptake was 51% and FIT positivity was 10%. The programme has undertaken over 400 screening colonoscopies and detected 154 precancerous adenomas and 38 cancerous lesions. CONCLUSIONS: The first round of The Adelaide and Meath Hospital Tallaght/Trinity College Dublin Colorectal Cancer Screening Programme has been highly successful and confirmed that there is an advantage for FIT based two stage bowel cancer screening programmes.

Office Ophthalmology: What is Urgent?

The author describes eye problems in general practice, from problems appearing in neonates to those in the elderly. Refractive problems are most common. Visually threatening eye disease that requires urgent treatment or referral is important, despite its rarity. Early detection and treatment of amblyopia is important in children. In diabetic retinopathy, a potentially blinding disease, maculopathy and proliferative retinopathy require immediate referral. Chronic simple glaucoma and cataracts cause gradual loss of vision in the older adult. Sudden loss of vision can be caused by optic neuritis, which is occasionally the first sign of multiple sclerosis. Retinal artery and retinal vein thrombosis are more common in older individuals with vascular disease. One of the most common causes of gradual visual loss in the elderly is senile macular degeneration.

Intense pulsed light therapy in the management of hereditary benign telangiectasia.

Hereditary benign telangiectasia is an idiopathic condition that causes considerable cosmetic embarrassment but has eluded successful treatment due to its extensive nature. We report a case of hereditary benign telangiectasia of the lower limbs in which the challenge of treating an extensive vascular lesion is met by using an Intense pulsed light (IPL) source.

Fumarate reductase of Escherichia coli: an investigation of function and assembly using in vivo complementation.

Recombinant plasmids which carried portions of the Escherichia coli frd operon were constructed and their expression examined by in vivo complementation of E. coli MI1443. This strain lacked a chromosomal frd operon and was unable to grow anaerobically on glycerol and fumarate. Introduction of all four fumarate reductase subunits into E. coli MI1443 was essential for the restoration of growth. The FRD A, FRD B dimer (but neither subunit alone) was active in the benzyl viologen oxidase assay. Both FRD C and FRD D were required for membrane association of fumarate reductase and for the oxidation of reduced quinone analogues. Introduction into E. coli MI1443 of the frdABC and frdD genes on two separate plasmid vectors failed to restore anaerobic growth on glycerol and fumarate. Thus separation of the DNA coding for the FRD C and FRD D proteins affected the ability of fumarate reductase to assemble into a functional complex.