Packing core-corona particles on a spherical surface.

We explore the non-trivial structures that can be obtained by the assembly of repulsive core-corona particles confined on a spherical surface. Using Monte Carlo simulations, we study the low-temperature equilibrium configurations as a function of the size of the confining (spherical) surface for a small number of particles (N ≤ 12) and obtain a large variety of minimal-energy arrangements including anisotropic and chiral structures. For a small cluster (N = 4), we construct a phase diagram in the confining surface radius vs corona range plane that showed regions where configurations with a certain energy are not accessible. Also, a phase diagram in the temperature and confining surface radius plane showed the presence of reentrant phases. The assembly of Platonic and Archimedean solids and the emergence of helical structures are also discussed. When the number of particles is large (N ≥ 100), apart from the appearance of defects, the overall configurations correspond closely to the ones formed in an unconfined two-dimensional case. Interestingly, the present model reproduces the symmetry of experimentally obtained small clusters of colloidal spheres confined at the surface of evaporating liquid droplets which cannot be explained in terms of packing of hard spheres. Thus, our simulations provide insight on the role that the softness of the particles may have in the assembly of clusters of nanoparticles.

Peer review audit of non-specialist occupational physician reports.

BACKGROUND: With declining specialist occupational physician (OP) numbers, there is increasing recognition of the importance of non-specialist physicians in occupational health (OH) service delivery, yet to date, this physician group remains understudied and their competency requirements poorly understood. AIMS: To evaluate the quality of a sample of non-specialist OH reports and compare these with specialist reports. METHODS: A retrospective peer review audit of a convenience sample of 200 consecutive non-specialist and specialist OH reports from an Irish OH service using an assessment form based on the modified Sheffield Assessment Instrument for Letters SAIL(OH)1. RESULTS: Of the 200 peer reviewed OH reports, 159 (80%) were from non-specialists. For all questions, 87% and above of non-specialist reports were 'satisfactory' or 'above expected'. On the overall assessment, out of 10, the mean non-specialist report score was 6.8 (standard deviation (SD) 3-10) and the specialist score was 7.3 (SD 3-10). Comparatively, non-specialist reports highlighted legal/ethical issues marginally more and adhered slightly better to contractual/ethical/legal boundaries, while specialist reports fared better in addressing manager's questions, in their structure and clarity and in covering all significant aspects of the case, particularly if the case was complex. CONCLUSIONS: Our findings demonstrate a high standard of OH report quality in this sample of non-specialist OPs that is consistent across all key OH report components. Potential development areas are also identified that can inform education/training tailored to this physician group and assist in competency standard-setting.

Hybrid ADFT Study of the C104 and C106 IPR Isomers.

This work presents a hybrid auxiliary density functional theory (ADFT) study of the neutral and hexaanionic C104 and C106 fullerenes with the aim to determine their ground state structures. To this end, all C104 and C106 fullerene structures that obey the isolated pentagon rule (IPR) were optimized with the Perdew-Burke-Ernzerhof generalized gradient approximation followed by a single-point energy calculation with the PBE0 hybrid functional. Our studies show that this composite approach yields relative energies of giant fullerenes that are accurate to around 1 kcal/mol. As a result, the ground states of C104, C1046-, and C1066- can be assigned to the isomers 234:Cs, 821:D2, and 891:Cs, respectively. On the other hand, the energetically lowest lying IPR isomers of C106, 331:Cs, 1194:C2, 534:C1 are separated by less than 1 kcal/mol which makes an unequivocal ground state assignment by hybrid DFT methods impossible. To guide future experiments, we also report the simulated IR and Raman spectra of the most stable neutral and hexaanionic C104 and C106 fullerenes.

The role of intragestational ghrelin on postnatal development and reproductive programming in mice

The purpose of this study was to evaluate the intragestational role of ghrelin in offspring development and reproductive programming in a mouse model of ghrelin imbalance during pregnancy. Female mice were injected with ghrelin (supraphysiological levels: 4 nmol/animal/day), antagonist (endogenous ghrelin inhibition with (D-Lys3)GHRP-6, 6 nmol/animal/day) or vehicle (control = normal ghrelin levels) throughout the pregnancy. Parameters evaluated in litters were growth, physical, neurobiological and sexual development and, at adulthood, reproductive function. Litter size and initial weight did not vary between treatments. Male pups from dams treated with ghrelin showed higher body weight increase until adulthood (31.7 ± 0.8 vs control = 29.7 ± 0.7, n = 11­14 litters/treatment; P < 0.05). Postnatal physical and neurobiological development was not modified by treatments. The antagonist accelerated male puberty onset, evidenced as earlier testis descent and increased relative testicular weight (antagonist = 0.5 ± 0.0% vs ghrelin = 0.4 ± 0.0% and control = 0.4 ± 0.0%, n = 5­10 litters/treatment; P < 0.05). At adulthood, these males exhibited lower relative testicular weight and reduced sperm motility (63.9 ± 3.6% vs control = 70.9 ± 3.3 and ghrelin = 75.6 ± 3.0, n = 13­15 animals; P < 0.05), without changes in plasma testosterone or fertility. Female pups intragestationally exposed to the antagonist showed earlier vaginal opening (statistically significant only at Day 25) and higher ovarian volume (antagonist = 1085.7 ± 64.0 mm3 vs ghrelin = 663.3 ± 102.8 mm3 and control = 512.3 ± 116.4 mm3; n = 4­6 animals/treatment; P < 0.05), indicating earlier sexual maturation. At adulthood, these females and those exposed to ghrelin showed a tendency to higher percentages of embryo loss and/or foetal atrophy. In conclusion, ghrelin participates in reproductive foetal programming: alterations in ghrelin activity during pregnancy modified body weight increase and anticipated puberty onset, exerting (or tending to) negative effects on adult reproductive function.

Influence of the number of daily pills and doses on adherence to antiretroviral treatment: a 7-year study.

WHAT IS KNOWN AND OBJECTIVE: Antiretroviral treatment (ART) is hampered by complicated regimens, high pill burden, drug-drug interactions, and frequent short- and long-term adverse effects, leading to decreased adherence. Over recent years, considerable effort has been directed at developing regimens that are less burdening. We undertook a 7-year retrospective study of the records of 264 HIV-infected subjects enrolled in a pharmaceutical care programme to document the progress made and to study the influence of the number of ART pills and doses on the level of treatment adherence. METHODS: Antiretroviral dispensing records were analysed for the number of pills and doses administered and the ART adherence rate estimated. RESULTS AND DISCUSSION: In 2005, the patients took a mean of 6·2 pills daily (CI 95%: 5·9-6·6), and 92·9% of them were on a twice-a-day (BID) dosage regimen. By 2012, the mean number of pills was reduced to 4·1 (CI 95%: 3·8-4·4), and only 50·9% were on a BID regimen. No statistically significant relation was observed between number of daily pills and doses and ART adherence reached by the patients in any of the analyses performed. WHAT IS NEW AND CONCLUSIONS: There has been a continuous reduction in the number of pills and doses of antiretrovirals taken by individual patients over the last 7 years due largely to the introduction of improved treatments and regimens. More daily pills or doses was not associated with worse ART adherence in our pharmaceutical care programme.

Risk factors associated with hepatitis C virus infection in an urban population of the State of Mexico.

The aim of this study was to determine the seroprevalence anti-hepatitis C virus (HCV) antibodies and risk factors associated with patients attending primary-care clinics in the State of Mexico. A cross-sectional, prospective study was conducted on 10,524 consenting patients with history of at least one risk factor for HCV. Antibodies were detected by immunoassay, third-generation ELISA; RT-PCR was carried out to confirm HCV infection. The seroprevalence of HCV antibodies was 1.2% (128). The most common risk factor was blood transfusion prior to 1993 (56.3%), followed by family history of cirrhosis 29 (22.7%); tattoos and/or piercings, 28 (21.9%); high-risk sexual practices, 4 (3.1%); healthcare work, 8 (6.3%); and intravenous drug use, 1 (8%). RT-PCR was performed on samples from 83 patients. Forty-five were considered positive. Genotype 1a was the most prevalent (37.7%).

A structural and surface approach to size and shape control of sulfur-modified undoped and Fe-doped TiO2 anatase nano-materials.

In situ synchrotron X-ray diffraction and diffuse reflectance infrared spectroscopy (DRIFTS) are combined to study the influence of sulfur on the crystallization of pure and Fe-doped titania nano-materials. Using these two tools we have investigated the effect of sulfur on the nucleation and growth processes of the anatase polymorph from amorphous powders and show how the addition of sulfur controls the primary particle size and shape of the materials. As well known, sulfur leads to the stabilization of the oxide particle size against sintering during thermal treatments and here we interpret the physico-chemical basis of such behaviour as an exclusive effect on grain growth kinetics, in turn linked to the dehydration of the surface layers of the materials. In addition this work shows that the presence of sulfur also affects the shape of the anatase particles, favouring the existence of (101)-type surfaces and elongated (along the c crystallographic axis) particles. This combined analysis of how sulfur influences morphological aspects of the anatase phase as it grows provides a basis for understanding of surface and chemical properties of anatase nano-powders that are highly dependent upon particle morphology.

[Characterization of Vibrio cholerae non-O1 and non-O139 isolates associated with diarrhea]. / Caracterización de aislamientos de Vibrio cholerae no-O1, no-O139 asociados a cuadros de diarrea.

Vibrio cholerae, etiologic agent of cholera, is transmitted to humans by ingestion of contaminated food or water. Even though serogroups O1 and O139 are the ones usually associated to epidemic cholera, isolates from other serogroups also cause gastroenteritis and extraintestinal infections. During the period 2003-2005, presence of V. cholerae in stools was investigated in children with diarrhea that seaked assistance at the Niño Jesús Hospital in Tucumán. Thirty four isolates of V. cholerae non-O1, non-O139 were recovered. We characterized the isolates studying its virulence factors by PCR, antimicrobial susceptibility patterns and genetic diversity by pulsed-field gel electrophoresis. Eight virulence patterns were obtained although no isolate was positive for the cholera toxin or the thermostable toxin. Four isolates were positive for the type three secretion system. The 17.6% of the isolates were resistant or intermediate to ampicillin and 5.9% were resistant to trimethoprim-sulfamethoxazole. By Sfil-PFGE, all isolates were genetically very diverse, as 27 different patterns were identified in 29 typeable isolates by pulsed-field gel electrophoresis. Although it has a low incidence, V. cholerae continues to be a causative agent of diarrhea in children, who are affected by a variety of circulating strains of V. cholerae non-O1, non-O139.

[Phenotypic and genotypic characterization of imipenem-resistant Pseudomonas aeruginosa isolated in a Buenos Aires hospital]. / Caracterización fenotípica y genotípica de la resistencia a imipenem en Pseudomonas aeruginosa aisladas en un hospital de Buenos Aires.

From 129 P. aeruginosa isolated at a health care centre located in Buenos Aires (Hospital "Eva Perón"), 14% produced IMP-13. Although 18 isolates were metallo-beta-lactamases (MBL) producers, only those isolates that displayed altered outer membrane protein profiles correlated with the resistant category according to CLSI or even Subcomisión de Antimicrobianos, SADEBAC, AAM. Phenotypic screening of metallo-beta-lactamases proved to be appropriate for detecting MBL producing isolates. IMP-13 producing isolates corresponded to at least five different clonal types, which not only suggests the dissemination of the resistant strain but also of the resistant marker.

[Leucocyte esterase as a reliable diagnostic, cost-effective and fast in knee infections]. / Esterasa leucocitaria como prueba diagnóstica eficaz, económica y rápida ante un proceso infeccioso articular de rodilla.

BACKGROUND: The articular infection represents a challenge due to its complexity and its devastating effect when not treated promptly. We have various diagnostic studies: cultures, ESR, CRP, count of leukocytes, among others but none is specific, it takes more than 30 minutes to complete and require complex infrastructure. In this study we determine the sensitivity and specificity of the leukocyte esterase for detection of an infectious process joint in Mexican population. MATERIAL AND METHODS: From November 2015 to April 2016 was obtained synovial fluid from patients with diagnosis of knee infection with or without implant and without infection with degenerative pathology of the knee. It assessed the sample through the COMBI-SCREEN 11SYS leukocyte esterase with reading colorimetric test at two minutes determining positive for infection: two crosses, the remainder of the sample was sent to culture. RESULTS: We perform the test in 64 samples of synovial fluid of knee joint 19 diagnosed with infection and 45 without infection. Was obtained a sensitivity 100%, specificity of 88.24%, PPV 68.42% and PNV 100%, kappa index 0.753 using the program IBM SPSS Statistics 22, Python ver. 2.7. CONCLUSIONS: Leukocyte esterase is a fast, economical and effective to detect an infectious process against one inflammatory with high probability of success. This study showed an index of concordance 0.753 Kappa, proving to be reproducible so recommend be implemented in the emergency department at the national level.

ANTECEDENTES: La infección articular es un reto ortopédico por la complejidad diagnóstica y sus efectos devastadores al no tratarse oportunamente. Se cuenta con diversos estudios de diagnóstico: cultivo, VSG, PCR, conteo de leucocitos, entre otros, pero ninguno es preciso, tardan más de 30 minutos en realizarse y requieren una infraestructura compleja. En este estudio se determina la sensibilidad y especificidad de la esterasa leucocitaria para la detección de un proceso infeccioso articular en población mexicana. MATERIAL Y MÉTODOS: de Noviembre de 2015 a Abril de 2016 se obtuvo líquido sinovial de pacientes con diagnóstico de infección articular con o sin implante y sin infección con patología degenerativa de rodilla. Se evaluó la muestra mediante el test de esterasa leucocitaria COMBI-SCREEN 11SYS con lectura colorimétrica a los dos minutos, determinando positivo para infección: dos cruces, el resto de la muestra fue enviado a cultivo. RESULTADOS: Se aplicó el test a 64 muestras de líquido sinovial de rodilla, 19 diagnosticadas con infección articular y 45 sin infección. Se obtuvo una sensibilidad 100%, especificidad 88.24% VPP 68.42% y VPN 100%, índice de concordancia kappa 0.753 mediante el programa IBM SPSS Statistics 22, Python versión 2.7. CONCLUSIONES: La esterasa leucocitaria es una prueba rápida, económica y eficaz para detectar un proceso infeccioso contra un proceso inflamatorio con alta probabilidad de acierto. Este estudio presentó un índice de concordancia kappa de 0.753, demostrando ser reproducible, por lo que se recomienda implementarse en los servicios de urgencias a nivel nacional.

[Community-acquired methicillin-resistant Staphylococcus aureus infections in a hospital for acute diseases]. / Infecciones adquiridas en la comunidad por Staphylococcus aureus resistente a meticilina en un hospital de agudos.

Community-acquired methicillin-resistant Staphylococcus aureus infections in a hospital for acute diseases. Methicillin-resistant Staphylococcus aureus (MRSA) is one of the most prevalent pathogens associated with nosocomial infections. However, most recently, MRSA has arisen as an emerging community pathogen, causing serious infections, mainly among young patients. We herein describe 33 cases of infections caused by community-acquired MRSA (C-MRSA), diagnosed between May 2005 and June 2006, at "Eva Perón" Hospital. The isolations were retrospectively studied. Methicillin resistance was confirmed by means of the detection of the mecA gene, and the genes for two virulence factors (Panton-Valentine Leucocidin -PVL- and gamma-haemolysin) as well as the cassette mec type were screened by PCR. All the patients were previously healthy. Four patients under 12, presented bacteremia, one had serious pneumonia, and the three remaining patients had osteoarticular infections; all the patients over 12, had skin and soft tissue infections without systemic damage. The C-MRSA strains harboured cassette mec type IV, and the PVL and gamma-haemolysin genes. They were methicillin-resistant, with no other associated resistances. It is important to consider the presence of these community- acquired strains in order to develop strategies for their correct treatment.

[Psychological evaluation in patients treated with external fixator]. / Evaluación sicológica en pacientes tratados con fijador externo.

OBJECTIVE: To establish whether complications in treatment, readmissions and reinterventions are related to psychological disturbances. To determine if the type, age and time of use are related to psychological aspects. MATERIAL AND METHODS: Retrospective, observational and transversal study in patients treated with external fixator in the period from 2005 to 2010. We used clinical records, 2 statistical tests and Students t-test with the IBM SPSS Statistics program, version 24. RESULTS: Circular fixator: we observed a significant p of 0.024 in the relations between age and psychological alterations. Regarding the complications and the psychological alterations, we obtained a significant p 0.048. Monoplanar fixator: the difference was significant of 0.038 on the relation between readmissions and psychological alterations. DISCUSSION: There are psychological disturbances in patients and families that may interfere with the treatment and complications. The literature showed that if there are alterations in the initial questionnaire, they will remain equal and generate increased aggressiveness; if there were no initial alterations, there will be no changes. We observed less psychological effects in older patients (13-16 years) and greater anxiety in patients in the medium range (10-12 years), as well as in patients with residual deformities. CONCLUSIONS: The psychological intervention is important in the pre-, trans- and postsurgical time. Patients of medium age, which corresponds to preteens, presented more psychological disturbances; therefore, it is recommended to avoid the use of external fixators in this age range.

OBJETIVOS: Establecer si las complicaciones en el tratamiento, reingresos y reintervenciones están relacionadas con alteraciones sicológicas. Determinar si el tipo, la edad y el tiempo de uso se relacionan con los aspectos sicológicos. MATERIAL Y MÉTODOS: Estudio retrospectivo, observacional y transversal en pacientes tratados con fijador externo en el período comprendido de 2005 a 2010. Se utilizaron expedientes clínicos, pruebas estadísticas 2 y t de Student con el programa IBM SPSS Statistics versión 24. RESULTADOS: Fijador circular: se observó una p significativa de 0.024 en la relación entre la edad y las alteraciones sicológicas. En cuanto a las complicaciones en relación con las alteraciones sicológicas, se obtuvo una p significativa de 0.048. Fijador monoplanar: la p fue significativa de 0.038 en la relación de reingresos y las alteraciones sicológicas. DISCUSIÓN: Hay alteraciones sicológicas en los pacientes y familiares que pueden interferir con el apego y las complicaciones. En la literatura se observó que si en el cuestionario inicial hay alteraciones, éstas continuarán igual y generarán mayor agresividad; si no hubo alteraciones iniciales, no habrá cambios. Se identificaron menos efectos sicológicos en pacientes de mayor edad (13-16 años) y mayor ansiedad en pacientes del grupo medio (10-12 años), así como en aquéllos con deformidades residuales. CONCLUSIONES: Es importante la intervención de sicología pre-, trans- y postquirúrgica. Los pacientes de edad media, lo cual corresponde a preadolescencia, presentarán más alteraciones sicológicas, por lo que se recomienda evitar el uso de fijadores en este rango de edad.

[Leukocyte esterase as a diagnostic tool for an infectious disease of the knee]. / Esterasa leucocitaria como prueba diagnóstica ante un proceso infeccioso articular de rodilla.

BACKGROUND: Articular infection is an orthopedic challenge due to its difficult diagnosis and devastating results. Various diagnostic studies exist: culture, ESR, CRP, count of leukocytes, among others, but none is specific, they all take more than 30 minutes to complete, and require complex infrastructure. In this study, we determine the sensitivity and specificity of the leukocyte esterase for detection of an infectious process joint in Mexican population. MATERIAL AND METHODS: During November 2015 to April 2016, we obtained synovial fluid from two groups of patients: one with a diagnosis of synovial joint infection with or without implant, and the control group, without infection but with degenerative pathology of the knee. We evaluated the sample using the leukocyte esterase test COMBI-SCREEN 11SYS with colorimetric reading at two minutes; two crosses determined positive for infection; the remainder of the sample was sent for culture. RESULTS: We performed the test in 64 samples of synovial fluid, 19 diagnosed with articular infection and 45 without it. We obtained a sensitivity of 100%, specificity of 88.24%, PPV of 68.42%, and NPV of 100%; Kappa index of .753. CONCLUSIONS: Leukocyte esterase is an effective test to detect an infectious process against an inflammatory one with a high probability of success. This study presented an index of agreement Kappa of 0.753, proving to be reproducible.

ANTECEDENTES: La infección articular es un reto ortopédico por su complejidad diagnóstica y efectos devastadores al no tratarse oportunamente. Contamos con diversos estudios de diagnóstico: cultivo, VSG, PCR, conteo de leucocitos, entre otros, pero ninguno es preciso, tardan más de 30 minutos en realizarse y requieren de infraestructura compleja. En este estudio determinamos la sensibilidad y especificidad de la esterasa leucocitaria para la detección de un proceso infeccioso articular en población mexicana. MATERIAL Y MÉTODOS: Durante Noviembre de 2015 a Abril de 2016, se obtuvo líquido sinovial de pacientes con diagnóstico de infección articular con o sin implante, y de otros sin infección, con patología degenerativa de rodilla. Se evaluó la muestra mediante el test de esterasa leucocitaria COMBI-SCREEN 11SYS con lectura colorimétrica a los dos minutos; se determinó positivo para infección con dos cruces; el resto de la muestra fue enviado a cultivo. RESULTADOS: Realizamos el test en 64 muestras de líquido sinovial de rodilla, 19 diagnosticadas con infección articular y 45 sin infección. Se obtuvo una sensibilidad de 100%, especificidad de 88.24%, VPP de 68.42% y VPN de 100%; índice de Kappa de .753. CONCLUSIONES: La esterasa leucocitaria es una prueba eficaz para detectar un proceso infeccioso contra uno inflamatorio con alta probabilidad de acierto. Este estudio presentó un índice de concordancia Kappa de 0.753, con lo que demostró ser reproducible, por lo que recomendamos implementarlo en los servicios de urgencias a nivel nacional.

Nitric oxide donors induce calcium-mobilisation from internal stores but do not stimulate catecholamine secretion by bovine chromaffin cells in resting conditions.

The potential role of nitric oxide (NO) donors and peroxynitrites on both basal catecholamine (CA) secretion and modulation of calcium levels has been investigated in primary cultures of bovine chromaffin cells. NO donors did not modulate catecholamine secretion, while peroxynitrites induced a time dose-dependent increase in basal CA secretion. Two facts may explain the lack of these compounds on basal CA secretion. NO donors induce, on the one hand, an increase in intracellular calcium levels by depletion of internal IP3-stores from endoplasmic reticulum. On the other hand, a small calcium influx through N-type voltage-dependent calcium channels (VDCC), which seem not to be coupled to exocytosis of adrenaline and noradrenaline in chromaffin cells. Both effects, calcium-mobilisation from internal stores and calcium entry through N-type VDCC are mediated by cGMP synthesis. In contrast, peroxynitrites induce an increase in basal CA secretion by both a decrease of intracellular catecholamine content and a toxic effect on cellular membrane. All these results, taken together, could explain contradictory results in the literature on the role of NO on basal catecholamine secretion and on modulation of intracellular calcium in chromaffin cells.

SNAP, a NO donor, induces cellular protection only when cortical neurons are submitted to some aggression process.

Nitric oxide is a versatile molecule, which plays important physiological and pathological roles. Its protective and toxic actions have been already evidenced in several cell types. However, the protective effect in cortical neurons remains elusive. In this work, we demonstrate that the NO-donor SNAP may induce both neuroprotection and neurotoxicity in this sort of cells. The protective effect of NO was evidenced when cortical neurons were exposed to deleterious conditions, such as serum deprivation. Serum deprivation induces apoptotic cortical neuron death through a caspase-dependent mechanism. Under these conditions, SNAP was able to oppose cell death through both caspase-3 inhibition and/or increase of antiapoptotic protein levels (Bcl-2 and Bcl-x(L)). On the other hand, in a normally serum-supplemented medium, high dose of SNAP behaves as a neurotoxic agent, through a mechanism which involves caspase-3 activation.

SNAP, a NO donor, induces cortical neuron death by a mechanism in which the caspase pathway is implicated.

In this paper, we present data which demonstrate that, in cortical neurons, SNAP induces loss in cell viability as evaluated by the XTT test. This cell death started at 250 microM SNAP when the treatment was performed in a serum-free medium and at 10 microM when the treatment was given in the presence of serum. This death was mediated, at least in part, by an apoptotic mechanism detected by flow cytometry and DNA fractionation. The highest SNAP concentrations induced a dual behavior on caspase-3 activity. Concentrations of 250 microM in the absence of serum and 10 microM to 300 microM in the presence of serum produced caspase-3 activation. This indicates that NO induces neuronal death by an apoptotic mechanism in which the caspase pathway is implicated. Higher SNAP concentrations (500 microM to 1 mM) diminished the caspase-3 activity to levels similar or even lower than control values. This profile was observed in the absence as well as in the presence of serum in the medium. The caspase-3 inhibition mediated by the highest SNAP concentrations did not imply NO cellular protection since the caspase-3 inhibition mediated by these SNAP concentrations neither correlated with cellular viability nor with cellular apoptosis. The possible mechanism of caspase-3 inhibition at the highest SNAP concentrations used is discussed.

Experimental and computational models of neurite extension at a choice point in response to controlled diffusive gradients.

OJECTIVE: Axons are guided toward desired targets through a series of choice points that they navigate by sensing cues in the cellular environment. A better understanding of how microenvironmental factors influence neurite growth during development can inform strategies to address nerve injury. Therefore, there is a need for biomimetic models to systematically investigate the influence of guidance cues at such choice points. APPROACH: We ran an adapted in silico biased turning axon growth model under the influence of nerve growth factor (NGF) and compared the results to corresponding in vitro experiments. We examined if growth simulations were predictive of neurite population behavior at a choice point. We used a biphasic micropatterned hydrogel system consisting of an outer cell restrictive mold that enclosed a bifurcated cell permissive region and placed a well near a bifurcating end to allow proteins to diffuse and form a gradient. Experimental diffusion profiles in these constructs were used to validate a diffusion computational model that utilized experimentally measured diffusion coefficients in hydrogels. The computational diffusion model was then used to establish defined soluble gradients within the permissive region of the hydrogels and maintain the profiles in physiological ranges for an extended period of time. Computational diffusion profiles informed the neurite growth model, which was compared with neurite growth experiments in the bifurcating hydrogel constructs. MAIN RESULTS: Results indicated that when applied to the constrained choice point geometry, the biased turning model predicted experimental behavior closely. Results for both simulated and in vitro neurite growth studies showed a significant chemoattractive response toward the bifurcated end containing an NGF gradient compared to the control, though some neurites were found in the end with no NGF gradient. SIGNIFICANCE: The integrated model of neurite growth we describe will allow comparison of experimental studies against growth cone guidance computational models applied to axon pathfinding at choice points.

Molecular mechanisms of glutamate release by bovine chromaffin cells in primary culture.

Previous work indicated that glutamate could be involved in the regulation of catecholamine secretion in bovine chromaffin cells. Thus, the question arises on the source of this putative regulatory glutamate. In this work we have examined the possibility that glutamate could be released from chromaffin cells. Data from this study indicate that chromaffin cells are able to release glutamate when they are stimulated by different depolarising agents such as 60 mM KCl, 1 mM 4-aminopyridine or 50 microM veratridine. The amount of glutamate released by these compounds was 0.32 nmol/10(6) cells (9.24% of cellular glutamate content), 0.275 (7.86%) and 0.158 (4.52%) for KCl, 4-AP and veratridine stimulation, respectively. All these catecholamine-secretagogues induced glutamate secretion by two mechanisms: 1) a Ca(2+)-dependent, probably exocytotic, mechanism and 2) a Ca(2+)-independent mechanism mediated by reversion of the electrogenic glutamate transporter. Analysis of Ca(2+)-dependent and independent releases for different compounds carried out by several experimental approaches, indicate that Ca(2+)-dependent release was the predominant mechanism for release induced by 4-aminopyridine (84% of total release) and high KCl (63%) whilst Ca(2+)-independent release was predominant for veratridine (67%). The Ca(2+)-dependent glutamate release evoked by depolarisation of chromaffin cells with high KCl and 4-AP could be split into both a fast and a slow kinetic component, which might correspond to the release of docked and mobilised chromaffin granules, respectively. On the other hand, depolarisation of cells with veratridine result in glutamate release with only the fast kinetic component. In the case of 60 mM KCl-evoked glutamate release, the fast component exhibited a decay time of <1 s and accounted for 0.63 nmol glu/6x10(6) cells (70% of total exocytotic release), whereas the slow component, which exhibited a decay time of 231 s, accounted for the release of 0.27 nmol glu/6x10(6) cells (30% of total exocytotic release). By contrast in the case of 4-aminopyridine the fast component of exocytosis only represents a 19% of total secretion and the slow a 81% with a decay time of 94 s. These data are very similar to those found in neurones and support the possible intracellular origin of glutamate having a role in the regulation of catecholamine secretion from chromaffin cells. In support of this, we have found that glutamate secretion could be evoked by stimulation of the nicotinic cholinergic receptors.

Apoptosis and necrosis: two distinct events induced by cadmium in cortical neurons in culture.

(1) Cadmium is an extremely toxic metal commonly found in industrial workplaces, a food contaminant and a major component of cigarette smoke. Cadmium can severely damage several organs, including the brain. In this work, we have studied both the cadmium toxicity on rat cortical neurons in culture and the possible protective effect of serum. (2) Our results indicate that: (1) cadmium is taken up by the neurons in a dose and serum dependent way; (2) cadmium, at concentrations from 1 micro M or 10 micro M (depending on the absence or the presence of serum) up to 100 micro M, decreases the metabolic capacity, which was evaluated by the XTT (tetrazolium salt) test; (3) cadmium induces apoptosis and LDH (lactate dehydrogenase) release in a dose dependent way; (4) in a serum-free medium, the cadmium-induced apoptosis is accompanied by caspase-3 activation; (5) both the caspase-3 activation and the cadmium-induced apoptosis are reversed by N-acethyl-Asp-Glu-Val-Asp-aldehyde (Ac-DEVD-CHO), a selective caspase-3 inhibitor, indicating that the caspase-3 pathway is involved in cadmium-induced apoptosis in cortical neurons; and (6) the cadmium concentrations which produce caspase-3 activation do not modify the intracellular ATP levels; however, higher cadmium concentrations lead to both intracellular ATP depletion and ATP release, but do not increase the caspase-3 activity, indicating that cadmium also produces cellular death by necrosis. (3) These results suggest that cadmium induces either apoptosis or necrosis in rat cortical neurons, depending on the cadmium concentration.

Psychiatric diagnosis and intervention in older and younger patients in a primary care clinic: effect of a screening and diagnostic instrument.

OBJECTIVES: To determine whether patient age is associated with psychiatric diagnosis or provider intervention in a busy primary care clinic, and, if so, whether a screening and diagnostic tool, the PRIME-MD, modifies age-related differences. DESIGN, SETTING, AND PARTICIPANTS: PRIME-MD use, psychiatric diagnosis, and provider interventions for psychiatric conditions were recorded for eligible patients attending a Veterans Affairs Medical Center primary care clinic. Data from 952 younger (<65 years) and 1135 older patients (> or =65 years) were analyzed to determine whether there were age-related differences in diagnosis/intervention and if use of the PRIME-MD modified these differences. INTERVENTION: Implementation of the PRIME-MD, a two-step instrument consisting of a self-administered patient questionnaire and a provider-administered structured diagnostic interview. MEASUREMENTS: Outcome measures were rates of (1) PRIME-MD use, (2) overall psychiatric diagnosis, (3) new psychiatric diagnosis, and (4) provider intervention for psychiatric conditions. RESULTS: There was no association between patient age and PRIME-MD use. Older patients were less likely to receive a psychiatric diagnosis in analyses that adjusted for "highly positive" screening questionnaires (OR = .45; P<.001). Older patients were also less likely to receive an intervention for a psychiatric condition in analyses that adjusted for whether a psychiatric diagnosis (OR = .62, P = .015) or a new psychiatric diagnosis (OR = .36, P<.001) was made during the study visit. The PRIME-MD increased rates of diagnosis and intervention but did not alter age-related disparities. CONCLUSIONS: Decreased rates of psychiatric diagnosis and intervention in older primary care patients are of concern. Implementing the PRIME-MD will likely increase rates of diagnosis and intervention but will need to be accompanied by additional measures to eliminate age-related disparities.