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OBJECTIVE US allopathic medical schools have experienced improvements in racial and ethnic diversity among matriculants in the past decade. It is not clear, however, whether better representation of historically excluded racial and ethnic groups at medical school entry impacts subsequent stages of the medical training pipeline leading into a specific field. The aim of this study was to examine these trends as they relate to the neurosurgical medical education pipeline and consider the drivers that sustain barriers for underrepresented groups. METHODS Race and ethnicity reports from the American Association of Medical Colleges were obtained on allopathic medical school applicants, acceptees, and graduates and applicants to US neurosurgical residency programs from 2012 to 2020. The representation of groups categorized by self-reported race and ethnicity was compared with their US population counterparts to determine the representation quotient (RQ) for each group. Annual racial composition differences and changes in representation over time at each stage of medical training were evaluated by estimating incidence rate ratios (IRRs) and 95% confidence intervals (CIs) using non-Hispanic Whites as the reference group. RESULTS On average, Asian and White individuals most frequently applied and were accepted to medical school, had the highest graduation rates, and applied to neurosurgery residency programs more often than other racial groups. The medical school application and acceptance rates for Black individuals increased from 2012 to 2020 relative to Whites by 30% (95% CI 1.23–1.36) and 42% (95% CI 1.31–1.53), respectively. During this same period, however, inequities in neurosurgical residency applications grew across all non-Asian racialized groups relative to Whites. While the incidence of active Black neurosurgery residents increased from 2012 to 2020 (0.6 to 0.7/100,000 Black US inhabitants), the prevalence of White neurosurgery residents grew in the active neurosurgery resident population by 16% more. CONCLUSIONS The increased racial diversity of medical school students in recent years is not yet reflected in racial representation among neurosurgery applicants. Disproportionately fewer Black relative to White US medical students apply to neurosurgery residency, which contributes to declining racial representation among all active neurosurgery resident physicians. Hispanic individuals are becoming increasingly represented in neurosurgery residency but continue to remain underrepresented relative to the US population. Ongoing efforts to recruit medical students into neurosurgery who more accurately reflect the diversity of the general US population are necessary to ensure equitable patient care.

BackgroundIncreasing numbers of older patients are presenting to emergency departments(ED) following trauma. These patients require multidisciplinary care that the traditional trauma model fails to provide. A Silver Trauma Review Clinic(STRC) was developed in conjunction with the geriatric, ED and multidisciplinary services to improve the post-discharge care of patients with non-operative traumatic injuries.We aimed to assess the STRC by reviewing the journey and outcomes of patients who attended the clinic and examining new diagnoses and interventions.MethodsA retrospective review of electronic chart data was performed on all patients who attended the clinic over the initial 1 year period.Results137 patient were reviewed with a median age of 80(IQR 12.5), 69% female. The median clinical frailty scale was 3 with a median time from the patient’s initial ED presentation to clinic of 15 days(IQR 11.25) and median time from initial review to discharge 20 days(IQR 34). 71% of presentations were as a result of falls under 2 metres. Primary injuries were 34% vertebral fractures, 45% limb fracture, 18% thoracic trauma, 11% pelvic trauma with 15% of patients suffering from multiple injuries. Patients attending the STRC had a comprehensive geriatric assessment with abnormal Mini-Cog assessments found in 29%, a new diagnosis of osteoporosis in 43% and orthostatic hypotension diagnosed in 13% of patients. 61% were discharged to primary care, 19% linked into a specialist geriatric clinic.ConclusionThe STRC is a novel approach allowing timely, patient focused, comprehensive and collaborative trauma care of older patients following non-operative injuries.

Background The Department of Health published new guidance in November 2020 ‘Vitamin D advice for people aged 65 and older’ which states that all adults >65 years should be supplemented with 15micrograms of Vitamin D daily. TILDA research states that 55.5% of adults >65 years have either insufficient or deficient vitamin D levels in winter (TILDA 2020). The Frailty Intervention Team (FIT) assess frail adults >65 years in the Emergency Department using medication review and Comprehensive Geriatric Assessment (CGA) to identify vitamin D supplementation. The audit aim is to determine if the new guideline is being implemented. Methods All patients who had a medicine reconciliation completed by the FIT Pharmacist in January 2021 were included in a retrospective audit. Patients were identified from the pharmacists worklist. Data was collected from a Hospital Clinical System using an excel data collection sheet. The data recorded included: Variable Indicative of Placement risk (VIP), Age, Sex, Rockwood Clinical Frailty Score (CFS), serum vitamin D level, date of last serum vitamin D level and vitamin D supplementation. Results The FIT Pharmacist completed 60 medicine reconciliations in January 2021. 70% (n = 42) were prescribed vitamin D. Of those 92% (n = 39) were prescribed a supplement containing at least 15micrograms of vitamin D. 85% (n = 51) had a vitamin D level reported in the preceding 6 years. 76% (n = 39) had sufficient vitamin D levels (>50 nmol/L). 21% (n = 11) had insufficient or deficient vitamin D levels ( Conclusion The new guideline was implemented in 65% (n = 39) of frail patients >65 years. The rate of vitamin D insufficiency in this cohort was lower than TILDA research suggests.

Background Many older people are brought to the Emergency Department (ED) who do not require acute hospital admission but there are risks to these patients if this occurs. The Frailty Intervention Team (FIT) are an interdisciplinary team based in the ED including physiotherapy, occupational therapy, speech and language therapy, dietetics, pharmacy, advanced nurse practitioners, and a geriatric registrar and consultant. They assess frail older patients and suggest alternative care pathways to admission. We aimed to investigate the impact of FIT on admission avoidance, bed days saved and to obtain patient feedback on their experience. Methods Routinely collected data from May 2021 to April 2022 was reviewed retrospectively. An anonymous patient feedback questionnaire was posted to 40 patients randomised from the FIT worklist between January to May 2022. Results 2,025 Comprehensive Geriatric Assessments (CGA) were completed between May 2021 and April 2022. 38% percent of patients were discharged home, 45% of this number had follow-up arranged. 104 patients were transferred directly to an offsite bed, mostly rehabilitation. We estimate we avoided 51 admissions to the acute hospital per month, almost half of these were patients admitted to the hospital and, who we discharged to alternative care pathways or home. The average length of stay in April 2022 was 22 days – by avoiding 615 admissions between May 2021 and April 2022 we have saved 13,530 bed days at an estimated cost saving of almost €11 million. 15 completed questionnaires were returned. 73% were very satisfied with their experience. 87% felt the FIT team helped facilitate their discharge from ED. The main themes identified from open ended questions included thorough assessment, patient centred care and satisfaction with early intervention and discharge. Conclusion A Frailty Intervention Team is a cost effective and patient centred way of avoiding unnecessary admissions for older people presenting to the ED.

Background The Department of Health published new guidance in November 2020 “Vitamin D advice for people aged 65 and older” which recommended that all adults ≥65 years should be supplemented with 15micrograms of Vitamin D daily. 55.5% of adults ≥65 years have insufficient/deficient vitamin D levels in winter (TILDA 2020). The Frailty Intervention Team (FIT) assess frail adults ≥65 years in the Emergency Department using medication review and Comprehensive Geriatric Assessment to identify vitamin D supplementation. The audit aim was to determine if compliance with the new guideline improved since the initial audit in January 2021. Methods All patients who had a medicine reconciliation completed by the FIT Pharmacist in January 2022 were included in a retrospective audit. Patients were identified from the pharmacist’s worklist. Data was collected from a Hospital Clinical System using a Microsoft Excel®. The data recorded included: Age, Sex, Rockwood Clinical Frailty Score (CFS) and vitamin D supplementation. Results The FIT Pharmacist completed 66 medicine reconciliations in January 2022. The mean age (+/-SD) was 82.4 (+/-7.6) years.The male to female ratio was 1:1.75The median CFS was 5 (mildly/moderately frail) with an IQR of 1. 68% (n=45) were prescribed vitamin D. Of those 89% (n=40) were prescribed a supplement containing at least 15micrograms of vitamin D. 32% (n=21) were not prescribed any vitamin D supplementation. 8% (n=5) were prescribed doses lower than 15micrograms of vitamin D. Conclusion Compliance with the new guideline was 61% (n=40) in comparison to 65% (n=39) in the initial audit. The results are reflective of adherence to the new guideline in the community. The policy update and education provided post the initial audit has possibly influenced the prescribing of vitamin D in the acute setting which unless patients are re-presenting will not have been captured in the outcome of this re-audit.

Background Inappropriate polypharmacy and ‘Potentially Inappropriate Prescriptions’ (PIP) are associated with increased morbidity and hospitalisation, in particular among frail older persons. A structured medication review, in conjunction with a Comprehensive Geriatric Assessment (CGA), can address PIP. The aim of the study was to review Frail Intervention Team (FIT) pharmacist interventions for frail, older adults presenting to the Emergency Department (ED) and experiencing medication compliance difficulties, polypharmacy and PIP. Methods Patients identified for medication review included those experiencing polypharmacy or medication compliance issues, presenting with a fall, delirium and/or frailty syndromes or complex comorbidities. The medication review process involved completion of medicines reconciliation and medication appropriateness review in accordance with the 7-Steps Medication Review Model (Scottish Government Polypharmacy Model of Care Group, 2018). Pharmacist optimisation recommendations were reviewed by a Consultant Geriatrician or Registrar and discussed with the patient prior to implementation. Results The FIT Pharmacist completed medication reviews for 765 patients between May 2021 and April 2022. The mean age (+/-SD) was 83.1 (+/-7.0) years with a median Clinical Frailty Score (CFS) of 5 (mildly/moderately frail). Medication Optimisation recommendations were actioned in 63% (n=483) of patients reviewed. The most commonly encountered PIP’s included: excessive anti-hypertensive/diuretic therapy, long-term acid-suppression therapy, anticholinergics and long-term prophylactic antimicrobials. Prescribing opportunities identified included: bone protection, laxatives and pain management. Conclusion FIT pharmacist review, in conjunction with the CGA, led to medication optimisation interventions in the frail older adult cohort presenting to the ED. Future studies should examine the impact of medication review on patient outcomes post-discharge.

Background The Clinical Frailty Scale (CFS) is a widely used frailty measurement that is recommended for use as part of a Comprehensive Geriatric Assessment. The Variable Indicative of Placement (VIP) score was developed to identify at triage in the emergency department patients who require geriatric input. There have only been a limited number of previous studies that have validated the VIP score for use at triage. Current practice in our hospital is to use the VIP score at triage to identify frail older patients and to use the CGA as part of the CGA performed by the Frailty Intervention Team (FIT). We aimed to assess the validity of the VIP tool to identify frail, older patients as identified using the CFS. Methods A retrospective analysis was performed using routine data collected for the month of November 2021. Inclusion criteria were patients over the age of 65 who had both a VIP and CFS performed. We used a CFS of 1-4 to define fit patients and 5-9 to define frail patients. Similarly for the VIP score we defined fit as VIP negative and frail as VIP positive. Statistical analysis was performed using Microsoft Excel. Results 267 patients were identified as meeting the inclusion criteria. 157 patients were found to be positive for frailty using the VIP score and 123 using the CFS. The sensitivity of the VIP score was 75% and the specificity was 55%. The positive predicative value was 59% and negative predictive value was 72%. Conclusion The VIP score fails to perform well in the real world as a screening tool for frailty in comparison to the CFS.

Higher Education Students (HES) may find the lectures as something far away from their everyday life. Professors feel a big gap between themselves and their pupils. Furthermore, some students can find that subjects are not connected in between them and make close boxes of knowledge. To show them the great net of knowledge they can build, we have redesigned an old strategy, the use of blogs in education, to break that gap and increase student interest in the lectures. After the introduction of the Bologna Process in the European Higher Education Area, annual subjects have almost disappeared in behalf of shorter ones. This reduction might help the students pass their courses, but it brings up the problem of compartments in knowledge. Because of this, our project has been focused on the students of the Pharmacy Faculty which have 4 subjects associated to infectious diseases (ID) from 3rd to 6th semester (Microbiology, Parasitology, Immunology, and Biological Analysis) of a curriculum of 5 years (10 semesters). Technological gap is a reality in between students and teachers because the internet and technology are used in a different way. HES have grown in a world where social media is a part of them, sharing their daily life with pictures, videos or comments. We have therefore thought to use this enthusiasm to create a blog to share and associate the knowledge they acquire in our classes, with news they find related to ID. The blog is edited by teachers, but written by students named the “Infectious Gazzete”. Students have looked for information related to ID in different media. Once they have found attractive news, students have to practice their critical thinking and write a small blog entry containing a review of the subject using their lectures notes or books. By means of this activity, students can link the knowledge acquired in the lectures to the latest news in the field of ID. Before the entry is published in the blog, teachers play an editor role where the entry is sent back to students for improvement. Finally, the text is uploaded to the blog. As the blog entries are published in an open blog, their peers are able to read and discuss the news. Because this activity is carried out during 4 semesters, students are also able to have a global and up to date view of every aspect related to ID. Furthermore, students from other Universities (The Montfort University, UK) have joined our initiative contributing to the blog. A survey was carried to analyze students’ opinion on this initiative. Ninety-five students answer the survey with a mean age of 19,11 (± 2,11) years old. After statistically analyzing the results, the medians that favorably scored for the activity were for the statements: In general, I liked this activity. The realization of this activity is interesting and I consider that this project favors my Personal development. None negative impression was observed for any question. Overall our project has improved students’ interest in the different subjects that could be used to interconnect the different topics addressed in the lectures. Moreover, the use of the blog as an open platform, make students feel the responsibility of being well informed and read about a topic before open publishing. The exposure of students to this experience creates a platform where we introduce our subjects into an open discussion in students’ families and friends, increasing the attention and interest of infectious diseases in their everyday life.

Resumen Objetivo Evaluar la utilidad del aporte de concentrado de medula osea rico en celulas troncales mesenquimales (MSC) como tratamiento de la necrosis avascular de cabeza femoral (NAV) para evitar o retrasar la artroplastia total de cadera (ATC). Material y metodos Estudio clinico unicentrico, prospectivo, no controlado, de pacientes con diagnostico de NAV. Evaluacion de parametros demograficos, Harris Hip Score (HHS), pruebas de imagen (radiografia y RMN); estadificacion mediante escala de Arlet-Ficat. Seguimiento a 1, 6, 12 y 24 meses. Aspirado de medula osea de cresta iliaca, concentrado con sistema Harvest SmartPReP 2, e infusion en zona de necrosis mediante forages. Resultados Veintidos caderas en 17 pacientes fueron reclutados entre los anos 2006-2012, seguimiento minimo 2 anos. Extraccion media de 119,5 ml de aspirado, implantandose 15,25 ml de concentrado de MSC. Durante los primeros 2 anos de la infusion 5 caderas (24,7%) precisaron ATC, sin diferencias en estadio de Ficat basal, y 4 (80%) de ellas presentaban afectacion de cabeza femoral igual o superior al 30%. En las 17 caderas restantes, se observo a los 2 anos de la infusion un aumento significativo del HHS (14,27) (p = 0,026) y disminucion del dolor (0,98) en la EVA (p = 0,089). Nuestros resultados sugieren que la infusion de concentrado de medula osea rico en MSC asociado a descompresion quirurgica del nucleo mejora significativamente la funcionalidad de la cadera (HHS) y evita la ATC en el 75,3% de los pacientes con NAV tratados durante los primeros 2 anos.

Objective To assess the efficacy of implanting concentrated bone marrow rich in mesenchymal stem cells (MSC) for the treatment of femoral head avascular necrosis (AVN) to prevent or delay total hip replacement (THR). Material and methods A single-centre, prospective, non-controlled clinical study was conducted on patients with a diagnosis of AVN. The parameters assessed were, patient demographics, Harris Hip Score (HHS), imaging test (X-ray and NMR), and staging using the Arlet–Ficat scale. The patients were followed up for 1, 6, 12 and 24 months. The bone marrow was aspirated from the iliac crest, concentrated with the Harvest SmartPReP 2 system, and infused into the necrotic area by means of core decompression. Results A total of 22 hips in 17 patients were recruited between the years 2006 and 2012, with a minimum follow-up of 2 years. A mean of 119.5 ml of aspirate was extracted, with 15.25 ml of MSC being implanted. During the first 2 years of the infusion, 5 hips (24.7%) required THR, with no differences in the baseline Arlet–Ficat stage, and 4 of these (80%) had femoral head involvement equal to or higher than 30%. A significant increase of 14.27 ( P = .026) in the HHS and a 0.98 ( P = .089) decrease in the VAS was observed two years after the infusion in the remaining 17 hips. The results suggest that the infusion of concentrated bone marrow rich in MSC, combined with surgical decompression of the nucleus, improves hip function (HHS), and avoids THR in 75.3% of patients with AVN treated during the first 2 years.

Background Sublingual immunotherapy has been proven as a well-tolerated and effective treatment for allergic rhinitis. Within this type of treatment, GRAZAX® is the most documented product in terms of safety and efficacy. The objective of this study was to identify the patients’ expectations and level of treatment satisfaction, as well as the clinical management of patients with moderate/severe allergic rhinoconjunctivitis treated with GRAZAX®. Methods This was a non-interventional, observational, multi-centre, open-label study involving a total of 131 adult patients aged 18–66 years with confirmed diagnosis of grass-allergy and initiated treatment with GRAZAX® between June 2010 and April 2011. Results In the pollen season after starting treatment, 56.6% of patients stated that their symptoms were much less/less intense, 86% needed less symptomatic medication for control of their symptoms, and 74.4% manifested to have improved (quite/a lot) as regards their allergic disease since treatment was initiated as compared with previous grass pollen season. The patient satisfaction with GRAZAX® was measured using a visual analogue scale (VAS) between 0 (minimum satisfaction) and 100 (maximum satisfaction) comprising five different items: effectiveness, tolerability, cost, convenience and overall satisfaction. The results obtained for each item were [mean (SD)]: 74.7 (18.1), 70.3 (36.1), 39.3 (25.8), 86.2 (12.6), 78.4 (15.8) respectively. The patient’s level of satisfaction is highly influenced, especially in terms of assessment of effectiveness, tolerability and convenience, by the information provided by the specialist. Conclusions In summary, it can be concluded that improved communication leads to increased patient knowledge, greater patient compliance, and increased patient satisfaction.

Dislocation after total knee arthroplasty is a rare complication and a difficult problem to address. When the flexion gap is larger than the extension gap and the collateral ligaments are injured, instability and knee arthroplasty dislocation can occur.We report the case of a patient presenting with a posterior dislocation of a posterior-stabilized prosthesis without trauma. Frank instability in varus stress test and a positive anterior drawer test with tibial internal rotation dismissed the conservative treatment.A constrained condylar prosthesis was used for the revision. He suffered a similar episode after a month, which demonstrated that the increase in the level of constraint was not enough to correct the severe asymmetric instability in flexion due to the damaged external structures. A rotating-hinge prosthesis was then implanted and the patient reported no additional episodes of instability.We made an exhaustive review of the literature, analyzed the possible causes that can lead to the tibiofemoral instability after a total knee arthroplasty and described some technical considerations.La luxación tras la artroplastía de rodilla es una complicación poco frecuente y de difícil manejo. Una brecha en flexión demasiado grande asociada a una laxitud de los ligamentos colaterales puede llevar a la inestabilidad y a la luxación en flexión.

Poly(2,2′-dioxy-1,1′-biphenyl)phosphazene (I) is a weak base that can be reversibly protonated with HBF4 in acetone. Acidic degradation of poly(2,2′-dioxy-1,1′-biphenyl)phosphazene was experimentally performed using HBF4, HCl, and H2SO4 yielding the same polymer I with lower molecular weights. The reaction takes place through protonation of some of the skeletal bonds thus producing a protonated P+-NH bond, and posterior rupture of those bonds by reaction with water. The hydrolysis breaks only these protonated skeletal bonds, leaving unaffected the regular P−N bonds. The degree of hydrolysis is controlled by the amount of time during which the system was treated with the acid. Molecular dynamics simulations performed on systems containing protonated and unprotonated oligomers together with water and HCl molecules proved that Cl− ions and water molecules get close enough to protonated P+−NH bonds as to produce a hydrolytical rupture, however, they do not get into such a proximity to the regular P−N bonds. T...

Anisakis simplex crude extracts (CE) (IPI, ASAC and ALK-ABELLÓ), A. simplex larval antigens purified using a column of IgG anti-A. simplex (PAK) or a column of IgG anti-Ascaris suum (PAS), antigen eluted from columns of IgG anti-A. suum (EAS) and an A. suum adult CE were assayed by the skin prick test. Thirty percent of assayed patients showed a negative reaction in the Anisakis skin prick test. Of 70% positives, two patients had a weal greater than that produced by histamine with the A. simplex extract from ABELLÓ and IPI. The A. suum skin prick test was positive in 35% of patients, with a lower reaction than that observed with the A. simplex extract from IPI in 57% of the sera and a higher reaction in 28% of the sera. All patients with positive reactions with the crude extract also showed positive weals with the two purified antigens, PAK and PAS. All patients, except three, with a reaction to A. suum antigen, were positive to the EAS antigen. In five patients the weal size produced by PAS was greater than that observed with PAK, whereas in another six patients the contrary was observed. Only one of these six patients did not react to EAS antigen, coincident with the patient showing only a slight increase (7%) in the weal size induced by PAK vs. PAS. When the EAS antigen was tested on patients positive to both PAK and PAS, six patients presented a weal size of >30% and only three patients who were positive to PAS did not react to the EAS antigen. These three patients were also negative against the A. suum CE. Purification by affinity chromatography eliminates from the PAS antigen the proteins responsible for producing cross-reactions with Ascaris (present in the EAS antigen).

Size exclusion chromatography with dual detection, i.e., employing a refractive index, concentration sensitive, detector together with a multiangle light scattering detector which is sensitive to molecular size, has been applied to study the solution properties of guar gum in water with different concentrations of K2SO4 at 25 °C. The analysis of a single highly polydisperse sample is enough for obtaining calibration curves for molecular weight and radius of gyration and the scaling law coefficients. The influence of the ionic strength on the conformational properties of the polymer can also be analyzed. Moreover, unperturbed dimensions can be obtained by extrapolation of the values measured in a good solvent. The value of the characteristic ratio of the unperturbed dimensions thus obtained is Cn=〈r2〉0/nl2≈19±1. A theoretical analysis is also included. Thus, molecular dynamics procedures were employed to analyze the conformational properties of an oligomer of guar gum under different conditions; namely, st...

Size exclusion chromatography (SEC) with differential refractive index (RI), multiangle light scattering (MALS) and UV detectors has been used to obtain information about copolymers of styrene (S) and methyl methacrylate (MMA). Homopolymers PS and PMMA having broad molecular weight distributions, mixtures of PS and PMMA homopolymers, and copolymers containing different weight fractions of S and MMA units were studied. Molecular weight distributions, molecular dimensions and scaling laws are reported for these systems. The behavior of the random copolymers is very different from that exhibited by the heterogeneous mixtures of homopolymers. Thus, the results obtained for the copolymers can be explained by assuming that these molecules have homogeneous distributions of S and MMA units along their chains. On the contrary, the mixtures produce the results that would be expected from heterogeneous combinations of different molecules. The SEC technique offers the possibility of determining the composition of random copolymers and discriminating among random copolymers (having homogeneous distributions of units) and block copolymers.

Solution properties of poly(N-vinylcarbazole) (PVCz) and copolymers of N-vinylcarbazole with methyl methacrylate are investigated by using SEC coupled with differential refractive index (RI), multiangle light scattering (MALS), and UV detectors. The technique affords information on molecular weight, molecular weight distributions, and the chemical composition of copolymers. Moreover, the polymer dimensions are investigated in two different solvents, tetrahydrofuran (THF) and toluene, which are good and poor solvents, respectively, for poly(N-vinylcarbazole). The collapse of the polymer chain, the scaling laws, unperturbed dimensions, and characteristic ratios are determined by employing only a broad polydisperse polymer sample. The results are in good agreement with those obtained by much more complex procedures for the homopolymer PVCz. The influence of the methyl methacrylate content in the dimensions of the copolymer chains is analyzed. Unperturbed dimensions are also calculated employing a simple two-...

Size-exclusion chromatography with coupled multiangle light scattering and differential refractometry detectors has been used to obtain molecular mass and radius of gyration distributions of polydisperse polymer samples. From these data the scaling law between dimensions and the absolute molecular mass is obtained with just one sample of each polymer. Three different kinds of polymers are presented: polystyrene which serves as reference polymer, polyphosphazenes which behave abnormally in solution and poly(ethylene oxide) which is soluble in water. Since the relationship between dimensions and molecular mass depends on the extent of interactions between chain segments and solvent molecules, the scaling law provides information about the solution properties of the polymer.

Introduction Rhodopseudomonas palustris is a purple nonsulfur anoxygenic phototrophic bacterium that is ubiquitous in the environment. R. palustris is of great interest due to its high metabolic diversity. R. palustris is capable of producing hydrogen gas, making it a potential biofuel producer. We have created a mutant variant (strain 2044) which is derepressed for nitrogenase biosynthesis in the presence of ammonia, enabling copious quantities of hydrogen gas to be produced. The wild-type strain was incapable of producing hydrogen under similar conditions. The biological goal of this project was to characterize and compare the proteomes of wild-type versus strain 2044 under photoheterotrophic conditions. This study enabled comparison of technical and biological replicates with label free quantitative “shotgun” proteomics. Methods Triplicate batches of R. palustris Strain 2044 and CGA010 wild-type were grown to log phase under photoheterotrophic conditions with ammonia present. Cells from each biological replicate were harvested, washed with buffer, and disrupted with sonication and fractionated by centrifugation. Proteome fractions were denatured, reduced, digested with trypsin, desalted, and concentrated. All samples were analyzed via a two-dimensional nano-LC-ES-MS/MS (2-dimensional linear ion trap mass spectrometer operating in data-dependent MS/MS mode). All MS/MS spectra were searched with SEQUEST and filtered/compared with DTASelect/Contrast. The three biological replicates were analyzed with duplicate technical replicates for each fraction. Label free quantitation was preformed by comparing relative protein abundance by sequence coverage, spectral count, and peptide count between WT and Strain 2044 proteins by statistical methods. Preliminary results R. palustris Strain 2044 was constructed by knocking out the two RuBisCO genes. Initially this double mutant did not grow photoheterotrophically; however after a lag of about 21 days; growth was obtained under these conditions. This culture was then streaked to isolate single clones capable of growth under photoheterotrophic conditions without a lag, which yielded strain 2044. This strain 2044 was subsequently shown to produce copious quantities of molecular hydrogen under photoheterotrophic growth in the presence of high levels of ammonia. Our goal was to determine up and down-regulation of proteins important to this new mutant and its potential for hydrogen gas production. We prepared individual peptide solutions from each replicate proteome for duplicate technical replicates by LC/LC-MS/MS on a linear ion trap. Our technical goal was to compare the variability between instrumental technical replicates and the true biological variability in living cell cultures. From the first biological replicate we identified 2,385 proteins with at least two unique peptides with conservative filtering parameters. From the WT strain 2045 proteins were identified with 80% reproducibility. From the mutant strain 2032 proteins were identified with 78% reproducibility. Over 150 proteins showed dramatic up or down expression. Many proteins involved in nitrogen fixation, such as NifX, NifK, NifD, FixA, FixB, FixC and FixX, were highly expressed in the mutant strain, even though fixed nitrogen was readily available in the media. The biological replicates are currently being analyzed with technical duplicates. Initial results have indicated that biological variability appears to be more substantial than technical variability. Our newly developed statistical package for label free quantitation (Wang et al. submitted JPR, Jan. 2005) was used to compare results across all biological and technical replicates. Novel Aspect : First systematic characterization of biological versus technical variability in a hydrogen-producing derepressed mutant of R. palustris.

En: Parasitology research. eISSN 1432-1955. vol. 103, 2008, págs 713-715. A majority of Kudoa spp. infects the somatic muscle of fish establishing cysts. Previously, elevated humoral responses were detected in BALB/c mice immunised with Kudoa sp. pseudocyst extracts and in BALB/c mice orally inoculated with Kudoa sp. pseudocysts, as well as the presence of anti-Kudoa sp. antibodies in human sera by enzyme-linked immunosorbent assay. The objective of this work was to test Kudoa sp. pseudocyst extracts by the skin prick test. Fifteen patients with gastroallergic and/or allergic symptoms related to fish ingestion were examined. Kudoa sp. pseudocyst extracts were administered (1 mg/ml) on the volar forearm skin. Four of the 15 selected patients were positive to Kudoa sp. extracts. The saline solution negative control did not induce any reaction.

Between 1985 and 1996, our Service treated 18 cases of osteochondritis dissecans of the talus in children and adolescents. The lesion is more frequent during childhood than previously thought. Different theories about the etiology of the lesion and the various treatments used are discussed. The outcome was satisfactory in most cases. We consider that, with the exception of type IV Berndt and Harty lesions, preliminary treatment should be conservative, which gave good results in our study. Surgical treatment should be reserved for patients with an unsatisfactory evolution with orthopaedic treatment, with lesions with thick sclerotic edges, or for patients with loose intraarticular fragments.

SummaryThrombin interacts with its receptor and thrombomodulin on endothelial cells. We evaluated the respective roles of these two proteins on human umbilical vein endothelial cell (HUVEC) growth by comparing thrombin, S195A (a mutant thrombin in which the serine of the charge stabilizing system had been replaced by alanine), and the receptor activating peptide (TRAP). Thrombin and TRAP induced DNA synthesis (half maximal cell proliferation with 5 nM and 25 μM, respectively), whereas S195A thrombin was inactive, inferring that growth is mediated through the thrombin receptor. Surprisingly, cells stimulated by TRAP exhibited a maximal proliferation twice greater than that obtained with thrombin. Combination of thrombin and TRAP resulted in a mitogenic response higher than by thrombin alone, but lower than by TRAP alone. The role of thrombomodulin was evaluated by adding an anti-thrombomodulin antibody, which prevents formation of the thrombin-thrombomodulin complex. Antibody did not interfere with cell proliferation induced by TRAP, but enhanced that induced by thrombin. We conclude that formation of the thrombin-thrombomodulin complex restrains HUVEC proliferation mediated through the thrombin receptor.

This study aimed to determine the cause of acute recidivous urticaria in patients who usually eat fish or other seafood. Twenty-five patients were studied. The skin prick test with larval Anisakis simplex extract was performed; total and specific IgE against A. simplex was measured with the CAP System; specific antibodies to A. simplex were determined by ELISA; and immunorecognition patterns of the sera were studied by Western blot. Nineteen patients showed specific IgE to A. simplex, but specific IgE to Ascaris was demonstrated in only two patients. No patients reacted to Toxocara canis or Echinoccocus granulosus antigens with the same test. The skin prick test was positive in 16 patients, in two of them persisting for 48 h. Five patients showed neither skin reaction nor specific IgE to A. simplex. Sera showed specific immunoglobulin levels against A. simplex larval crude extract, by both ELISA and Western blot. Likewise, specific immunoglobulin levels against excretory-secretory antigen were also measured by ELISA. Only one patient showed sensitization to fish. A. simplex was found to be the main cause of acute recidivous urticaria in patients who usually eat fish and are not sensitized to it.

We report the effects of aqueous extracts of six marine microalgae on responses to various contractile agents in isolated rat duodenum (acetylcholine, calcium chloride, barium chloride) and vas deferens (noradrenaline). Our results suggest that only extracts of Phaeodactylum tricornutum, Skeletonema costatum, and Chlorella stigmatophora may have pharmacological potential. Extracts of these three species had spasmolytic effects on responses to the three contractile agents in duodenum. In vas deferens, P. tricornutum and C. stigmatophora inhibited the contractions induced by noradrenaline, whilst S. costatum showed a non-dose-dependent potentiation. With the aim of elucidating their mechanisms of action, we are attempting to isolate the active principles present in these extracts.

This paper describes advances in the development of several electrooptic shutter devices utilizing the quadratic electrooptic effect of lead lanthanum zirconate titanate (PLZT) ceramic wafers. Aperture sizes utilized in these PLZT devices ranged from 25 microm to 0.25 m. Practical applications of the shutters discussed in this paper include eye protection in military and industrial applications, a goggle-type device with dual synchronously operated PLZT shutters for use in a stereoscopic three-dimensional TV display, an electrically controlled variable density filter for use with vidicon tubes, a large-aperture photographic shutter for image motion compensation cameras, and a page composer for use in a holographic memory system.

We report the effects of aqueous extracts of the microalga Skeletonema costatum Greve (Cleve) (Bacillariophyceae) on spontaneous motor activity, rectal temperature, motor coordination, amphetamine-induced hypermotility, exploratory behaviour, muscle relaxation, catalepsy, conditioned avoidance responses, oxotremorin-induced cholinergic syndrome, and pentylenetetrazole-induced convulsions. The S. costatum extract at dosages of 310 and 620 mg/kg works like an antidopaminergic drug with anticholinergic properties, that does not induce catalepsy and with a notable muscle relaxing activity.

We report the effects of aqueous extracts of the microalgae Dunaliella tertiolecta Butcher (Chlorophyceae) and Phaeodactylum tricornutum Bohlin (Bacillariophyceae) on oxotremorin-induced cholinergic symptoms, amphetamine-induced hypermotility, pentylenetetrazole-induced convulsions, and Rota-rod test performance. Both extracts significantly reduced rectal temperature, spontaneous motor activity, and time on the Rota-rod and increased the number of mice falling off the rod before 180 s, but neither protected against pentylenetetrazole-induced convulsions or against oxotremorin-induced tremor, salivation, and diarrhoea.

Using Western blot techniques, the specificities of crude and purified (PAK and PAS) Anisakis simplex antigens were compared against 24 sera from patients diagnosed with Anisakis sensitization. All patients recognized a 60 kDa protein against the A. simplex crude extract, while 37.5% and 12.5% reacted with proteins of 40 and 25 kDa, respectively, when IgG was tested. In the case of IgE determination, 41.6% of sera were negative, while 12.5% and 20.8% appeared to cross-react against Toxocara canis and Ascaris suum, respectively. When the PAK antigen (A. simplex antigen purified by means of a column of IgG anti-A. simplex) was tested, immune recognition towards the 60, 40 and 25 kDa proteins increased in 83.3%, 16.7% and 4.2%, respectively, when the Ig antibodies were tested. In the case of the PAS antigen (PAK antigen purified by means of a column of IgG anti-A. suum), the reaction against the 40 and 25 kDa proteins increased to 45.8% and 25%, respectively, when Ig antibodies were used. Finally, when the EAS antigen (eluted from the anti-A. suum column after PAK purification) was tested, 83.3% of the assayed sera reacted against the 14 kDa protein, when the Ig antibodies, IgG and IgM immunoglobulins were measured. With the IgE determination, the reactions were observed in 41.7% of patients with proteins between 60 and 35 kDa against the PAS antigen. With the EAS antigen, reactive bands of 184, 84 and 14 kDa appeared. In conclusion, in the purification process of the A. simplex larval crude extract, the proteins implicated in cross-reactions with Ascaris and Toxocara were eliminated, with an important concentration of proteins responsible for the induction of specific responses.

Parkinson's disease (PD) is a neurodegenerative disorder, characterised by the progressive loss of dopaminergic neurons in the substantia nigra, and typically treated by dopamine replacement. This treatment, although very effective in the early stages of the disease, is not curative and has side-effects. As such there has been a search for a more definitive treatment for this condition, which has mainly concentrated on replacing the lost neurons with neural grafts. Possible cell sources for replacement range from autologous grafts of dopamine secreting cells to allografts of fetal ventral mesencephalon and neural precursor cells derived from fetal tissue or embryonic stem cells. Some of these cells have been the subject of clinical trials, which to date have produced disparate outcomes. Therefore, whilst cell therapies remain a promising treatment for PD, there is need for further refinement of the techniques involved in this experimental procedure, before any new trials in patients are undertaken.

One of the fundamental aspects of a parasitic infection diagnosis is the use of adequate antigens to develop specific and sensitive immunoassays. This fact is especially complicated in nematode infection cases because of the high cross-reactivity among different parasites in this group. We performed an evaluation of Anisakis simplex antigens purified by affinity chromatography. We used sera from 38 patients diagnosed with Anisakis sensitization and sera from 35 patients with clinical suspicion of visceral larva migrans (VLM). These sera were assayed by the ELISA method against the crude extracts (CEs) and the purified antigens. When the sera from patients diagnosed with Anisakis sensitization were tested against the A. simplex CE, the IgG was the most abundant immunoglobulin. When the A. simplex larval antigens were purified using a column of IgG anti-A. simplex (PAK) or a column of IgG anti-Ascaris suum (PAS) were tested, we observed a higher diminution in the IgG levels, which coincides with the augmentation of the mean values against the "eluted of Ascaris" (EAS antigen). When the IgE was detected, only 18.4% of the sera reacted with the PAS antigen. We have observed that in the purification process of A. simplex antigen by affinity chromatography, the majority of the proteins that produced cross-reactivity against A. suum and Toxocara canis were eliminated.

En: Acta Tropica. ISSN 0001-706x. vol. 89, 2003, págs 85-89. The aim of this work was to study cross-reactivity in the diagnosis of two related ascaridosis. Nineteen patients diagnosed with recidivous acute urticaria (RAU) caused by Anisakis simplex and 26 patients diagnosed with visceral larva migrans (VLM) caused by Toxocara canis were studied employing commercial diagnostic kits and “in house” assay kits. Cross-reactivity observed was greater when using “in house” assay kits, suggesting that T. canis excretory–secretory antigens were not only recognized by antibodies from patients with RAU but with greater intensity compared to the A. simplex excretory–secretory antigens.

The main objective of this paper is to present a simple, general methodology for tracing the power output of each generator, on an individual basis, throughout the electrical power network. The tracing algorithm is based on power flows as given by a load flow solution of the power network. The contribution of each generator to the reactive power flowing in each element of the network becomes readily available. Accurate information as to which generator supplies which load and, in what proportion, is also part of the solution. The veracity and simplicity of the method is demonstrated by a numeric example.

The purpose of this study was to determine whether 99mTc-mercaptotriacetylglycine (MAG3) can substitute for 99mTc-glucoheptonate (GH) in the detection of pyelonephritis.One hundred thirty renal scintigraphies were evaluated retrospectively in 38 children (21% boys, 79% girls; age range 1 mo-21 yr; mean age 7.2 yr) referred for evaluation during an acute clinical urinary tract infection and for follow-up studies. Twelve topographical regions were designated on each kidney. Each area was graded for severity of decreased radionuclide localization: mild (Grade 1), moderate (Grade 2) or marked (Grade 3). Early posterior views of MAG3 studies were compared to delayed posterior GH images. In all patients, both studies were performed on the same day.Eighty-two studies were performed during an acute clinical infection and 48 were performed as follow-up. Seventy-seven percent of the studies had focal cortical lesions. Of all the cortical lesions identified by GH, MAG3 detected 74% (match lesions). A comparable percentage of lesions was identified in each region by both studies. GH scintigraphy detected 261 lesions (63 Grade 1, 149 Grade 2 and 49 Grade 3), and MAG3 detected 201 lesions (37 Grade 1, 117 Grade 2 and 47 Grade 3). MAG3 was unable to recognize 60 lesions identified by GH studies in 11 patients (mismatch lesions). Of these, 41% (26 of 63) were Grade 1, 21% (32 of 149) were Grade 2 and 4% (2 of 49) were Grade 3. In three cases, MAG3 identified lesions not seen by GH (reverse mismatch); all had acute symptomatic infection.These data document that MAG3 in the early phase of the study (1-2 min) can detect Grade 2 to Grade 3 cortical lesions in patients with pyelonephritis, but it is less effective in detecting Grade 1 lesions.

Dehumidifiers (DH) are potentially effective appliances as coadjuvant therapy in the treatment of bronchial asthma caused by sensitization to house dust mites. The aims of this study were to analyze DH tolerance in asthmatic patients, to assess the parasitological effects and to analyze the environmental effects produced by the use of these appliances in the bedrooms of asthmatic patients sensitized to house dust mites. 10 stable asthmatic patients sensitized to house dust mites were studied. DH appliances (CD-300) were installed in their bedrooms. Each patient was given symptom scoring tables and a portable peak expiratory flow (P.E.F.) during a period of 5 months, 1 month before installing the DH and 4 months afterwards. To study the parasitological efficacy of the DHs, we analyzed dust samples from the bedrooms and determined the Der p I, Der f I and Der II allergens by means of a modified ELISA based on monoclonal antibodies. Dust samples were collected before installing the DHs and after they had been working for 2 and 4 months. Dry temperature and relative humidity measurements at three time intervals (7-9, 15-17 and 22-24 h) were carried out. The 1st measurement was done prior to installation of the DHs in the patients' bedrooms and the 2nd and 3rd were achieved 2 and 4 months respectively after the installation. Statistical analysis was done by comparison of paired means. No significant differences were detected in the patients' symptoms nor in the P.E.F. measurements in the course of the study. Decreases in the house dust mite allergens were observed in 4 bedrooms. A significant decrease in relative humidity in the bedrooms of mite asthma patients after use of dehumidifier appliances was observed (p0.01). Significant differences between the measurements of the bedrooms with and without DH were detected (p0.01). In summary, DHs were well tolerated by stable asthmatic patients, produced a significant decrease in the relative humidity level and showed some parasitological efficacy.

Eosinophils are important effector cells in allergic inflammation described in allergic rhinitis (AR) and allergic bronchial asthma (BA). During the pollen season serum levels of eosinophil cationic protein (ECP) and eosinophil X protein/eosinophil-derived neurotoxin (EPX/EDN) are increased in BA. The aim of the present study was to evaluate the serum levels of ECP and EPC in pollen atopic patients with AR and BA during the winter. 92 patients were studied. They were divided into three groups: I 29 patients with AR, II 51 patients with BA and III 12 healthy subjects. Allergic rhinitis and bronchial asthma were diagnosed by routine clinical tests: clinical history, skin tests, total IgE and specific IgE. In addition ECP and EPX were determined in serum. All patients were asymptomatic, stable and without medical treatment. Methacholine challenge test (MCT) was performed in all patients. MCT were positive in 4 patients of group I and 45 patients of group II. ECP levels (ug/l) were: 21 (I), 24 (II) and 7 (III). EPX levels (ug/l) were 35 (I), 45 (II) and 21 (III). Statistical differences (p0.01) were observed both in ECP and EPX levels in patients with MCT positive in relation to patients with MCT negative, and in allergic patients (I and II) in comparison with the healthy subjects (III) (p0.01). ECP and EPX serum levels are increased in patients with a positive MCT in the winter, out of the pollen season, when patients are asymptomatic, stable and without treatment. This fact suggests that eosinophils play an important role in the pathogenesis of bronchial asthma.

We report on the case of a 32-year-old atopic patient who showed a severe anaphylactic reaction due to the ingestion of a pollen compound prepared in an herbalist's. A few minutes after ingestion, generalized pruritus, difuse erythema, facial edema, cough, hoarseness and dysphonia appeared, and the emergency administration of subcutaneous epinephrine and intravenous methylprednisolone was necessary. Skin tests with a battery of inhalants and food allergens were performed. The patient only showed sensitization to Artemisia vulgaris, Taraxacum officinalis and Salix alba. Specific IgE levels were evaluated by FEIA-CAP giving a seric level of CAP class 3 to Artemisia vulgaris and class 2 to Taraxacum officinalis and Salix alba. Samples of the pollen compound were shown in the microscopical analysis to be 93% pollens and 6% fungi. In the qualitative study Taraxacum officinalis (15%), Artemisia vulgaris (5%) and Salix alba (15%) were the main elements identified. In summary, this case study describes a food-induced systemic reaction due to a pollen compound in an atopic patient with a history of allergic rhinitis. Pollinic patients must be informed on the risks that the consumption of these compounds might cause.

In the course of our continuing pharmacological studies of marine microalgae, we look now at Tetraselmis suecica Stewart (Prasinophyceae) and Isochrysis galbana Demart (Haptophyceae), assessing the effects which aqueous extracts of these algae exert upon the central nervous system. To this end, we have tested for effects upon the following: spontaneous motor activity, body temperature, motor coordination, dextroamphetamine-induced hypermotility, exploratory conduct, muscle relaxation, catalepsy, conditioned avoidance response, oxotremorine-induced cholinergic syndrome, and pentylenetetrazole-induced convulsions. Our results show that aqueous extracts of both microalgae display action profiles which prove difficult to categorize by standard pharmacological criteria.

At the level of the central nervous system an aqueous extract of the marine microalga Chlorella stigmatophora (Butcher) (Chlorophyceae) showed anti-dopaminergic-type activity in assays of its effects on spontaneous motor response, rectal temperature, motor coordination, amphetamine-induced hypermotility, exploratory conduct, muscle relaxation, catalepsy, conditioned avoidance responses, oxotremorine-induced cholinergic syndrome, and pentylenetetrazole-induced convulsions.