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Purpose: To review the clinical and research value of optical coherence tomography angiography (OCTA) in the field of neurology., Methods: Current literature involving OCTA were reviewed through PubMed using the search terms "optical coherence tomography angiography", with "multiple sclerosis", "Alzheimer's disease", "optic neuropathy", or other closely-related terms., Results: OCTA has been applied in research to advance our understanding of the pathobiology of neurological disorders. OCTA-derived blood flow and vessel density measures are altered in multiple sclerosis (MS), Alzheimer's disease (AD), and various optic neuropathies (ON) in varying regions of the posterior segment vasculature of the eye. These emerging research findings support the occurrence of retinal vascular alterations across a host of neurological disorders and raise the possibility that vasculopathy can be clinically relevant since it contributes to the pathobiology of several neurological disorders., Conclusion: OCTA may be beneficial for neurological research. Additional investigations using OCTA in neurological disorders will help to further validate its clinical and research utilities in terms of characterizing the role of vasculopathy in neurological disorders., Competing Interests: Not applicable.The authors declare that they have no competing interests.

Introduction: In the short time since 2014, three pivotal, worldwide studies in neuromyelitis optica spectrum disorders have been launched: eculizumab, SA237 and inebelizumab, each based on a unique mechanism., Areas Covered: In this review, we provide a discussion on the trial data available for each drug, a brief description of the trial design, and our expert opinion on the potential benefits and risks., Expert Opinion: Eculizumab, a C5 complement inhibitor, may prove useful in the treatment of intractable cases of NMOSD, but physicians must be aware of the known risk of meningococcal infection. SA237, an interleukin-6 receptor blocker, may be effective at reducing relapse risk, and also has the potential to reduce neuropathic pain in NMOSD. Inebelizumab, a B cell depleting agent, has never been tested in NMOSD, but based on extensive evidence of efficacy with B cell depletion using rituximab, inebelizumab is expected to work at least as well.

Purpose: To compare the efficacy of Dr Organic Tea Tree Face Wash, OcuSoft Lids Scrub Plus and the BlephEx™ device at treating of Demodex folliculorum blepharitis., Methods: Eighty-six subjects (33 males/36 females) were enrolled in a randomised controlled interventional treatment study. Subjects completed a dry eye symptom questionnaire and were assessed for presence of Demodex folliculorum. Subjects were divided into three groups according to treatment: Dr Organic Tea Tree Face Wash (A) (n=28), OcuSoft Lid Scrub Plus (B) (n=30), or in-house lid scrub with the BlephEx™ device before nightly lid scrubs with OcuSoft Lid Scrub Plus (C) (n=28). Subjects were advised to clean their eyelids nightly for four weeks. Each subject was re-assessed for symptoms and Demodex folliculorum blepharitis after two weeks and four weeks of treatment., Results: The quantity of Demodex folliculorum was significantly reduced after four weeks of treatment in all three groups (p<0.05). Overall, there was no difference in efficacy between the three treatments (p>0.1). Symptoms reported by subjects were significantly improved after two and four weeks of treatment (p<0.05). Overall, there was no difference in efficacy between the three treatments to reduce symptoms after two or four weeks (p= 0.813 and p=0.646 respectively)., Conclusion: All three methods tested have shown good ability to reduce Demodex folliculorum quantity, improve subjective symptoms and help treat Demodex folliculorum blepharitis., (Copyright © 2017 British Contact Lens Association. Published by Elsevier Ltd. All rights reserved.)

The Universal Design for Learning (UDL) framework emphasizes multiplicity of representation, expression and engagement to cater for the widest possible set of learning styles and abilities. Digital Game Based Learning (DGBL) can slot into a universally designed approach to education as one of several alternative ways of learning that will suit some learning preferences, such as those who prefer to learn in an active way. However, DGBL can itself encapsulate the principles of UDL if the game designer embraces UDL as a fundamental set of game design principles. This paper discusses, with examples, the ways in which a game designer can universally design a DGBL solution with respect to game mechanics, representation and personalization, with an emphasis on the use of gameplay data for formative and summative evaluation during the design, build and retrospective phases, as well as for adaptive learning and formative feedback during the delivery phase.

Objectives: The aim of this paper was to critically review the recent literature on psychosocial and behavioral outcome following childhood stroke, to assess whether quality of life is affected and to investigate the factors related to psychosocial outcome., Methods: Literature searches were conducted and identified 36 relevant papers from the period 1908-2016., Results: The systematic review found that many children experience difficulties in a wide range of psychosocial domains. Quality of life can also be significantly reduced. Inconsistent findings regarding the correlates of outcome are likely due to methodological limitations. These issues include small and heterogeneous samples, lack of control groups, and measurement difficulties., Conclusions: This systematic review strongly indicates that childhood stroke can affect a myriad of psychosocial domains and a child's quality of life. Methodological issues, particularly around heterogeneous samples and measures, limit the conclusions that can be drawn regarding the predictors of outcome.

Objectives: Childhood stroke is a rare but devastating occurrence. Its infrequency has meant that a clear body of knowledge has not been fully established regarding its impact on neuropsychological outcome. Our aims were (i) to critically review the recent literature on neuropsychological outcome following childhood stroke; (ii) to investigate the factors related to neuropsychological outcome following childhood stroke., Methods: Literature searches were conducted and revealed 39 relevant papers from the period 1999-2015., Results: The review found that a significant number of children experience difficulties in a wide range of neuropsychological domains, with particular vulnerabilities noted in attention, speed of information processing and executive functioning. There were inconsistent findings regarding the correlates of neuropsychological outcome, which is likely due to methodological limitations of the studies., Conclusions: This review strongly indicates that childhood stroke can affect a myriad of neuropsychological domains, with attention, speed of processing and executive function particularly vulnerable. Methodological issues, particularly around heterogeneous samples and measurement difficulties, limit the conclusions that can be drawn regarding the predictors of outcome.

Personalised treatment of heart disease requires an understanding of the patient-specific characteristics, which can vary over time. A newly developed implantable surface acoustic wave pressure sensor, capable of continuous monitoring of the left ventricle filling pressure, is a novel device for personalised management of patients with heart disease. However, a one-size-fits-all approach to device sizing will affect its positioning within the pulmonary artery and its relationship to the interrogating device on the chest wall on a patient-specific level. In this paper, we analyse the spatial orientation and morphology of the pulmonary artery and its main branches in patients who could benefit from the device and normal controls. The results could optimise the design of the sensor, its stent, and importantly its placement, ensuring long-term monitoring in patient groups.

Objectives: To evaluate the bone health and vitamin D levels of a cohort of patients with relapses of multiple sclerosis (MS) and to propose an algorithm for the management of bone health in this patient group., Methods: We prospectively studied 56 consecutive patients from our acute relapse clinic. 3 patients were excluded from analysis as they were not deemed to have experienced an acute MS relapse. Bone health was assessed with vitamin D levels and Dual Energy X-ray Absorptiometry (DEXA) scanning (10 patients failed to attend for DEXA). Statistical analyses were used to compare groups and identify predictive variables. A review of the literature led to a proposed management protocol., Results: Pre-relapse the baseline EDSS was ≤6.5 in all subjects, and <4.0 in the majority (66%). Most received corticosteroids. 51% had low bone mineral density (BMD) as defined by a T-score less than -1.0 on DEXA scanning. Three were osteoporotic (T-score less than -2.5). Thirty one of fifty (62%) subjects were Vitamin D deficient (25(OH)D less than 50nmol/L). A range of variables, including previous corticosteroid usage, were not significantly predictive of reduced BMD., Conclusions: There was a high frequency of both low BMD and Vitamin D deficiency in this cohort of relatively young and largely ambulatory patients experiencing MS relapses. Current tools, such as the WHO FRAX algorithm, are inadequate in assessing bone status and fracture risk in this patient group, predominantly as they are focused on older age groups. We propose a simple clinical management algorithm., (Copyright © 2016 Elsevier B.V. All rights reserved.)

We describe the clinical and neuropathological features of two cases of cerebellar degeneration with selective involvement of the dentate nucleus. Both cases were associated with malignancy, however known paraneoplastic antibodies were absent. Pathological studies at autopsy confirmed T-cell-mediated neuronal destruction in the cerebellum which was strikingly limited to the dentate nucleus in both patients. The occurrence of these pathological features has not been previously described in antibodynegative paraneoplastic disease, but bears similarities to Rasmussen’s encephalitis.

The present article presents a method for assessing the radial stiffness of nitinol stents. An idealized stent model was created, and its radial stiffness was calculated by means of finite element modeling. The calculations were validated against experimental measurements. The variation of radial stiffness with geometrical dimensions was calculated, and the effect of increasing radial stiffness on endovascular deployment was analyzed. Peak tensile and compressive stresses as well as stent penetration were calculated in the case of an idealized pulmonary artery model having realistic dimensions as well as stiffness. The results of stress calculations were compared with a second set of simulations, where an idealized behavior of the stent (uniform expansion to a theoretical contact diameter) was modeled. The results show how in reality nitinol stents behave in a non-ideal way, having a non-uniform expansion and exerting non-uniform pressure on the contact areas with the artery. Such non-ideality decreases though with the increase in radial stiffness. The radial force alone may be insufficient in describing the stent-artery interaction, and numerical modeling proves to be necessary for capturing such complexity.

Purpose: The purpose of this paper is to examine the recording of clinical indication for prescribed sedative/hypnotic (SH) medications in a large, acute tertiary referral hospital., Design/methodology/approach: All hospital inpatients' medications (n = 367) were audited for prescription details regarding SH medications. Medical notes were then examined for evidence of a recorded indication for such medications., Findings: SH medications were prescribed to many hospital inpatients (42.5 per cent) during admission. An indication was documented in the nursing or medical records for 24.4 per cent of patients who were prescribed such medications. Nurses rather than by doctors prescribed most SH medications (74 vs 26 per cent, respectively, p = 0.003). Some patients receiving SH medications were both over 65 and impaired in their mobility (19.2 per cent). The treatment indication was documented in 47 per cent., Practical Implications: Most patients prescribed SH medications have nothing in their medical record explaining why these drugs are being used, including half of the elderly, less mobile patients. All health professionals dealing with SH medications and doctors in particular need to justify the use of such medications in the medical record. For the particularly high-risk groups where SH medications are potentially more dangerous, explicit guidance on why and how such medications are to be used must be provided by prescribers., Originality/value: For the first time, data are presented on documentation rates for clinical indication of prescribed SH medications across a large acute hospital, and highlights significant shortcomings in practice. This study should inform other organisations of the need to be mindful of facilitating greater compliance with good prescribing practice.

Purpose: Patients undergoing breast-conserving surgery were offered boost radiotherapy with targeted intraoperative radiotherapy (TARGIT) using the Intrabeam system to test the feasibility, safety, and efficacy of the new approach., Methods and Materials: We treated 302 cancers in 301 unselected patients. This was not a low-risk group. One-third of patients (98/301) were younger than 51 years of age. More than half of the tumors (172, 57%) were between 1 cm and 2 cm, and one-fifth (62, 21%) were >2 cm; 29% (86) had a Grade 3 tumor and, in 29% (87), axillary lymph nodes contained metastasis. After primary surgery, 20 Gy was delivered intraoperatively to the surface of the tumor bed, followed by external-beam radiotherapy (EBRT), but excluding the usual boost., Results: The treatment was well tolerated. The follow-up ranged from 3 to 80 months (164 and 90 patients completed 2 and 3 years follow-up, respectively). Four patients (1.3%) had local recurrence. The Kaplan-Meier estimate of local recurrence is 2.6% (SE = 1.7) at 5 years. This compares favorably with the 4.3% recurrence rate in boosted patients from the EORTC boost study, in which only 8.1% patients were node-positive, as opposed to 29% in our series., Conclusion: Targeted intraoperative radiotherapy combined with EBRT results in a low local recurrence rate. This could be attributed to both accurate targeting and timeliness of the treatment. These data support the need for a randomized trial to test whether the TARGIT boost is superior to conventional external boost, especially in high-risk women.

It is widely believed that reducing antimicrobial usage should reduce resistance, although observational evidence is mixed. Pneumococci make ideal subjects to test this belief as they are widely surveyed and lack an animal reservoir. Accordingly, susceptibility data for pneumococci in the UK and Ireland were retrieved from the Health Protection Agency's LabBase/CoSurv system and from the European Antimicrobial Resistance Surveillance System (EARSS) and British Society for Antimicrobial Chemotherapy (BSAC) databases. The BSAC surveillance examines respiratory pneumococci; the other systems focus upon invasive organisms only, with the LabBase/CoSurv system being the most comprehensive, capturing data on most bacteraemias in England and Wales. National pharmacy sales data were obtained from the IMS Health MIDAS database and were modelled to the resistance data by logistic and linear regression analysis. All systems except for the BSAC respiratory surveillance data indicated that penicillin resistance has fallen significantly since 1999 in the UK, whereas macrolide resistance has been essentially stable, or has risen slightly. The data for Ireland were based on smaller sample sizes but suggested a fall in penicillin non-susceptibility from 1999 to 2004, with conflicting evidence for macrolide resistance. The recent decreasing trend in penicillin resistance is in contrast to a rising trend in England and Wales until (at least) 1997 and strongly rising macrolide resistance from 1989 to 1993. UK pharmacy sales of macrolides and oral beta-lactams fell by ca. 30% in the late 1990s following increased concern about resistance, before stabilising or rising weakly; sales in Ireland were stable or rose slightly in the study period. We conclude that falling penicillin resistance in pneumococci followed reduced sales of oral beta-lactams to pharmacies in the UK, but a similar fall in macrolide sales was not associated with any fall in resistance. Stabilisation or decline in penicillin resistance has occurred in Ireland despite stable or increasing oral beta-lactam sales.

Objectives: We investigated the long-term efficacy of cryo ablation for treatment of atrial flutter., Background: To our knowledge, no study has assessed the long-term efficacy of cryo ablation by assessing both symptom and conduction recurrence., Methods: A total of 45 consecutive patients with symptomatic atrial flutter were ablated with a 7-F, 6-mm-tip, quadripolar cryo catheter (Freezor Xtra, CryoCath Technologies Inc., Kirkland, Canada). Electrophysiologic studies (EPS) were performed with diagnostic catheters. Cryo ablation was at -75 degrees C for 4 min, beginning at the inferior rim of the coronary sinus os and creating a posterior line to the Eustachian ridge. Safety, bi-directional isthmus block at intervention, and recurrence at three, six, and nine months post procedure were assessed., Results: There were no adverse events reported. All patients were free of discomfort on cryo energy delivery. The acute success rate at intervention was 87%. Follow-up data from 39 acutely successful patients showed 27 (69%) without conduction recurrence on repeat EPS at three months, and none (0%) had symptom recurrence documented by Holter monitoring, electrocardiogram, and/or patient diary records at three, six, and nine months follow-up., Conclusions: Our experience with a new 7-F, 6-mm-tip, quadripolar cryo catheter yielded a fairly high success rate at intervention, an excellent safety profile, and a good chronic success rate in terms of symptom recurrence. However, further monitoring is necessary to determine whether all asymptomatic patients continue to remain free of symptoms, given the small pool of patients demonstrated to have conduction recurrence.

Many LDCs currently on the market are not digested in the upper gastrointestinal tract and become fermented in the large intestine. They possess physiological benefits similar to those of dietary fibre. For some of these materials the fermentation process is highly specialised and leads to the selective stimulation and growth of beneficial gut bacteria, e.g. bifidobacteria. These materials are described as prebiotics, which are defined as nutrients fermented in the large bowel that favour the growth of desirable large bowel microflora. This activity has been demonstrated for inulin and oligofructose. Two other carbohydrates with low digestibility that offer desirable physiological properties are resistant starch (RS) and polydextrose (PD). These 'functional benefits have led to considerable interest from the food industry leading to the use of these ingredients in the development of new 'healthy' products. This paper describes the use of these materials in the development of 'healthy' products, some of their functional properties, and the benefits they confer on different food systems.

The challenging nature of studies of membrane proteins has made it difficult to determine the molecular mechanism of transmembrane signaling. For the bacterial chemoreceptor family, there are crystal structures of the internal and external domains, structural models of the transmembrane domain, and evidence for subtle ligand-induced conformational changes, but the signaling mechanism remains controversial. We have used a novel site-directed solid-state NMR distance measurement approach, using (13)C(19)F REDOR, to measure a ligand-induced change of 1.0 +/- 0.3 A in the distance between helices alpha 1 and alpha 4 of the ligand-binding domain in the intact, membrane-bound serine receptor. This distance change is shown not to be due to motion of the side chain and thus is due to motion of either the alpha 1 or the alpha 4 helix. Additional distance measurements can be used to determine the type of backbone motion and to follow it to the cytoplasm, to test and refine current proposals for the mechanism of transmembrane signaling. This is a promising general method for high-resolution measurements of local structure in intact, membrane-bound proteins.

The control of hospital-acquired infection, in particular methicillin-resistant Staphylococcus aureus (MRSA) remains a challenge. Our hospital has established a purpose built 11-bed cohort unit with on-site rehabilitation for care of patients colonized with MRSA, in an attempt to improve their quality of care. Prior to the opening of this unit a number of concerns were voiced and the aim of this study was to address these. First, to establish if patient cohorting reduces the likelihood of successful decolonization, second, to evaluate the risk of staff colonization, and finally to see if successful environmental control of MRSA is possible.A patient database was established detailing patient demographics, infection rates, eradication and reacquisition rates. Staff screening was performed weekly, at the start of a period of duty. Sixty environmental sites were screened before unit opening, at 48h, six weeks and at six months. There were 88 admissions in the first six months; 62 patients were colonized with MRSA, and 26 patients (10 surgical, 16 medical) had MRSA infections. Twenty-three of 88 patients (26%) were successfully decolonized, which compares favourably with an eradication rate of 20% for the rest of the hospital. Twenty staff members participated in weekly screening. Five staff members colonized with MRSA were detected and all were successfully decolonized. Environmental control was achieved with a combination of a daily detergent clean and a once weekly clean with phenolic disinfectant. Our preliminary data suggest that, despite cohorting patients colonized with MRSA, with proper education and supervised cleaning protocols, it is possible to control environmental MRSA load, successfully decolonize patients and limit the risk of staff colonization., (Copyright 2000 The Hospital Infection Society.)

Despite improvements in survival rates, infection remains an important cause of morbidity and mortality following solid organ transplantation. Prevention of infection and, failing this, prompt diagnosis and treatment remain the cornerstones of management. During the peri-operative admission, when the level of immunosuppression is at its height, nosocomial infection accounts for the majority of infective morbidity. Although the measures taken to prevent nosocomial infection may vary, centres undertaking such procedures must ensure that strategies are in place to protect patients. The importance of basic infection control measures cannot be over-emphasised. In addition, appropriate prophylactic agents, rapid diagnostic techniques and the early institution of appropriate therapy are essential. As developments in this field advance, the epidemiology of infection will continue to change, demanding an ongoing assessment of preventative, diagnostic and therapeutic strategies.

We report two cases of ecthyma gangrenosum which occurred at sites of iatrogenic trauma. The first case developed due to metastatic seeding with Pseudomonas aeruginosa during an episode of septicaemia and the second case occurred as a primary skin lesion. Both required prolonged courses of antibiotics and one patient died. The different pathogenic mechanisms and outcomes associated with this condition are discussed.

Aeromonas species were isolated from specimens other than faeces from 59 hospital inpatients over a 15 year period. Of the isolates, 79.7% were regarded as clinically significant, with skin and soft tissues and blood cultures as the commonest sites of infection. Of the isolates, 52.5% were hospital-acquired, and 55.9% of patients had serious underlying disease. Community-acquired infections in previously healthy individuals accounted for only 13.6% of isolates. However unlike many other opportunistic infections, aeromonas infection was not closely associated with prior antibiotic therapy, nor was there a significant increase in the frequency of infection over the study period.

Background and Objectives: Physicians in the 1920s and 1930s began to treat patients with chronic pain syndromes using regional anesthetic techniques for both temporary and permanent block of pain pathways. The founding of the American Society of Regional Anesthesia (ASRA) in 1923 provided a unique venue for the dissemination of information concerning regional anesthesia for both surgery and chronic pain management., Methods: The growth of chronic pain management on a national basis was assessed by using the Quarterly Cumulative Index to the Medical Literature to trace the distribution of information on regional anesthesia. From the Minutes of Meeting of the American Society of Regional Anesthesia, presented papers and the discussion that followed were analyzed., Results: Inquiries into regional anesthesia during the 1920s and 1930s predominantly dealt with technique. For the first time pain papers were listed under the regional anesthesia heading in the Index. The papers presented at ASRA meetings during the period helped develop the use of regional anesthesia for both chronic pain management and surgical anesthesia., Conclusions: The ASRA was instrumental in bringing together physicians interested in regional anesthesia and pain management. During the 1930s physician anesthetists came to predominate as the organization's officers and members and helped translate the work of the ASRA into a part of the knowledge required to be a specialist physician in anesthesia.

1. An 'atypical' 5-HT2 receptor which is located on the endothelium of rat jugular vein has been described. In the present study we have further defined the nature of the 5-HT2 receptor subtype present in this preparation. 2. In experiments conducted in the presence of ketanserin to preclude involvement of 5-HT2 receptors, the mixed 5-HT2B/2C antagonist, SB 200646, acted as an antagonist of 5-HT at the endothelial 5-HT receptor (pA2 = 7.2). Yohimbine, which exhibits negligible affinity for rat 5-HT2C receptors but has high 5-HT2B receptor affinity, acted as a potent but non-surmountable antagonist (pA2 > or = 7.3) in rat jugular vein. Neither yohimbine nor SB 200646 affected endothelium-dependent relaxations induced by carbachol. 3. Mianserin also acted as a surmountable antagonist (pA2 = 7.3) and the 5-HT2B agonist, BW 723C86, acted as a potent partial agonist (pEC50 [95% C L], intrinsic activity +/- s.e. mean = 7.9 [7.6-8.3], 0.84 +/- 0.04). Responses to BW 723C86 were antagonized by SB 200646 (0.3 microM) yielding an 'apparent' pA2 [95% CL] of 7.03 [6.76-7.32]. 4. These data are consistent with the presence of 5-HT2B receptors mediating endothelium-dependent relaxation of rat jugular vein.

1. The present study was undertaken to isolate and characterize pharmacologically homogeneous populations of 5-hydroxytryptamine (5-HT) receptors from a possible mixed receptor population mediating concentration of the longitudinal muscle of rat stomach fundus. Our aim was to extend the pharmacological characterization of the 5-HT2B receptor which is reported to be expressed in this preparation. 2. To minimize spontaneous activity and any influence of circular muscle on the contractile response, narrow (1-1.5 x 20 mm) segments of mucosa-denuded longitudinal muscle were used. Under these conditions, blockade of monoamine oxidase with pargyline (100 microM for 15 min) caused a leftward displacement of concentration-effect curves for both 5-methoxytryptamine (5-MeO-T) and tryptamine. Neither pargyline nor a number of uptake inhibitors affected responses to 5-HT. 3. In pargyline pretreated preparations, the order of potency of a number of tryptamine analogues was as follows: 5-MeO-T > or = alpha-Me-5-HT > or = 5-HT > 5-carboxamidotryptamine (5-CT) > tryptamine > 2-Me-5-HT. In addition several ligands known to act as agonists at either 5-HT2A or 5-HT2C receptors including 1-m-chlorophenylpiperazine (m-CPP), Ru 24969, MK 212 and SCH 23390 were also agonists in rat fundus whilst sumatriptan, renzapride and 8-hydroxy-2-(di-n-propylamino) tetralin (8-OH-DPAT) were very weak or inactive. With the exception of 2-Me-5-HT and m-CPP, most agonists produced monophasic concentration-effect curves consistent with an interaction at a single site. High concentrations of 2-Me-5-HT evoked relaxations which were blocked by phentolamine (1 MicroM) suggesting an interaction with alpha-adrenoceptors. m-CPP often evoked biphasic concentration-effect curves with a second contractile phase which was insensitive to yohimbine at concentrations higher than required for antagonism of responses to 5-HT.4. LY 53857, methiothepin, methysergide, ritanserin and ICI 170809 were potent but non-surmountable antagonists of 5-HT in rat fundus. In contrast, several ligands behaved as surmountable antagonists with the following order of potency: rauwolscine >yohimbine = mesulergine > mianserin = SB 204070 >WY 26703 > SB 200646> pirenpirone> renzapride. DAU 6285, granisetron, spiperone, ketanserin,phentolamine and GR 127935 did not affect responses to 5-HT at concentrations up to 1 pM. The agonist and concentration independent profile of antagonism supported a single site interaction for both agonists and antagonists.5. We conclude that despite small differences concerning the enantiomeric selectivity and affinity of rauwolscine and yohimbine, the close pharmacological identity of 5-HT receptors in rat stomach fundus and the recently cloned 5-HT2B receptor is maintained. SB 200646, which demonstrates some selectivity for 5-HT receptors in rat stomach fundus, should provide a useful ligand for confirmation of this view and allow discrimination of 5-HT2B function both in vitro and in vivo.

The chemiluminescent detection methods described in this chapter have been successfully applied to the detection of plasmid DNA and genomic DNA in Southern and sequencing protocols. The high sensitivity and the simplicity of AMPPD are instrumental in making the chemiluminescent detection of DNA successful in hybridization assays. This detection technique has also been used to detect DNA in dot blots and in situ hybridization experiments as well as proteins in enzyme-linked immunosorbent assays (ELISAs) and Western blots.

A chemiluminescent Western blotting procedure and its application in assays for human transferrin and human immunodeficiency virus-I antibodies are described. The procedure is based on a chemiluminescent substrate, adamantyl 1,2-dioxetane aryl phosphate and alkaline phosphatase-labeled detection antibodies. Different membranes (polyvinylidene fluoride, nitrocellulose, nylon) and a proprietary membrane treatment agent (Nitro-Block) have been studied. This sensitive blotting procedure utilizing AMPPD, a polyclonal rabbit anti-transferrin:goat anti-rabbit IgG-alkaline phosphatase detection complex, and a PVDF membrane blocked with Nitro-Block permits the detection of 125 pg (1.6 fmol) of human transferrin. A novel 1,2-dioxetane substrate, CSPD, has also been evaluated.

A 33 year old woman with irregular menses due to endometrial sarcoidosis and a family history of sarcoidosis is described. Its relative increase in importance in the wake of the decline in the incidence of genital tract tuberculosis in Ireland is discussed.

We have used a chemiluminescent detection method to improve both the sensitivity and the speed of detection of human genes with oligonucleotide probes. A direct chemiluminescent substrate (AMPPD) was used in combination with an alkaline-phosphatase-labeled oligonucleotide probe to detect the human tissue of plasminogen activator gene by Southern blot analysis. X-ray exposures obtained after 4 h were comparable to those obtained after 7 days with a 32P-labeled oligomer. After 16 h, the signal was 12 times greater than the 32P signal. The detection of the single-copy tissue plasminogen activator gene in 0.25 micrograms of human genomic DNA (76,000 molecules) was achieved. The improved sensitivity obtained by chemiluminescent detection should increase the usefulness of oligonucleotide probes in the direct Southern analysis of human genetic disorders.

Metallic platinum was deposited at surfaces of intracellular photosynthetic membranes of whole cells of a cyanobacterium (blue-green alga). The deposited platinum particles acted as a catalyst for generation of hydrogen from photosynthetic decomposition of water in the absence of other exogenous electron transfer agents. This technique represents a means of placing metal catalysts in contact with intracellular structures of microorganisms.

We have coupled a chemiluminescent detection method that uses an alkaline phosphatase label to the genomic DNA sequencing protocol of Church and Gilbert [Church, G. M. & Gilbert, W. (1984) Proc. Natl. Acad. Sci. USA 81, 1991-1995]. Images of sequence ladders are obtained on x-ray film with exposure times of less than 30 min, as compared to 40 h required for a similar exposure with a 32P-labeled oligomer. Chemically cleaved DNA from a sequencing gel is transferred to a nylon membrane, and specific sequence ladders are selected by hybridization to DNA oligonucleotides labeled with alkaline phosphatase or with biotin, leading directly or indirectly to deposition of enzyme. If a biotinylated probe is used, an incubation with avidin-alkaline phosphatase conjugate follows. The membrane is soaked in the chemiluminescent substrate (AMPPD) and is exposed to film. Dephosphorylation of AMPPD leads in a two-step pathway to a highly localized emission of visible light. The demonstrated shorter exposure times may improve the efficiency of a serial reprobing strategy such as the multiplex sequencing approach of Church and Kieffer-Higgins [Church, G. M. & Kieffer-Higgins, S. (1988) Science 240, 185-188].

A new chemiluminescent Southern blot procedure offers molecular biologists a safe, ultrasensitive and rapid alternative to conventional 32P-based systems. This new DNA detection system, SOUTHERN-LIGHT, has been developed by Tropix, Inc. The luminescent signal is produced from a direct chemiluminescent substrate, disodium 3-(4-methoxyspiro[1,2-dioxetane-3-2'-tricyclo-[3.3.1.1 .3,7]decan]-4-yl) phenyl phosphate (AMPPD), which decomposes upon dephosphorylation with alkaline phosphatase. SOUTHERN-LIGHT is an ultrasensitive, rapid detection kit for use with membrane-bound DNA. It is the first test kit to incorporate AMPPD. It requires no specialized equipment and results can be conveniently imaged on instant film or x-ray film within 5-60 min of exposure.

Five cases of painful bone resorption from the distal clavicle following shoulder trauma are reported to demonstrate that the lesion is generally benign. Surgical excision is curative. The multiplicity of circumstances leading to clavicular osteolysis suggest that the balance between bone accretion and resorption is precarious in this region of the skeleton.

Cooked chicken was allowed to spoil in a normal kitchen refrigerator (variable temperature) and at a standard 4C. After 10 days' storage, bacteria were isolated from the chicken. It was found that the numbers of organisms at variable refrigeration temperature were tenfold higher than those at a uniform 4C. In an attempt to find the sources of contamination, swabs were made of different areas of the kitchen. Many of the bacteria isolated from the spoiled chicken, were also isolated from the kitchen environment. When pure cultures of organisms isolated from spoiled chicken were inoculated into sterile cooked chicken and held at 4C, the main spoilage organisms were found to be Pseudomonas putida and Aeromonas hydrophila, which were also isolated from the refrigerator where the chickens were stored in the kitchen. Aeromonas hydrophila was found in significantly high numbers on plates, cutting knives, chopping boards and cold water taps.