Towards Independent Particle Reconstruction from Cryogenic Transmission Electron Microscopy

Coronary heart disease is the single largest killer of Americans so improved means of detecting risk factors before arterial obstructions appear are expected to lead to a improvement in quality of life with a reduced cost. This paper introduces a new approach to 3-D reconstruction of individual particles based on statistical modeling from a sparse set of 2-D projection images. This paper introduces a new approach to 3-D reconstruction of individual particles based on statistical modeling from a sparse set of 2-D projection images. The method is in contrast to the current state of practice where reconstruction is performed via signal processing or Bayesian methods that use averaged images acquired from an ensemble of particles. As such, this new approach has its impetus in use for novel diagnostic tests such as LDL and HDL particle shape characterization. The approach is also expected to have uses in areas such as quality assurance for drug delivery nano-technologies and for general proteomic studies. The individual particle reconstruction algorithm is based on a hidden Markov model. Higher order Markov chain statistics, which are generated from the a priori model of the target of interest, can be derived from traditional methods such as single particle reconstruction and/or the underlying physical properties of the particle. By placing the reconstruction voxel space at a 45° angle to the projection image, 4-passes of the HMM processing can be performed from a single image. Reconstruction results from a simple model and a single projection image resulted in better than 98% reconstruction accuracy as compared to the original target

Coronary artery endothelial dysfunction is positively correlated with low density lipoprotein and inversely correlated with high density lipoprotein subclass particles measured by nuclear magnetic resonance spectroscopy.

OBJECTIVE: The association between cholesterol and endothelial dysfunction remains controversial. We tested the hypothesis that lipoprotein subclasses are associated with coronary endothelial dysfunction. METHODS AND RESULTS: Coronary endothelial function was assessed in 490 patients between November 1993 and February 2007. Fasting lipids and nuclear magnetic resonance (NMR) lipoprotein particle subclasses were measured. There were 325 females and 165 males with a mean age of 49.8+/-11.6 years. Coronary endothelial dysfunction (epicardial constriction>20% or increase in coronary blood flow<50% in response to intracoronary acetylcholine) was diagnosed in 273 patients, the majority of whom (64.5%) had microvascular dysfunction. Total cholesterol and LDL-C (low density lipoprotein cholesterol) were not associated with endothelial dysfunction. One-way analysis and multivariate methods adjusting for age, gender, diabetes, hypertension and lipid-lowering agent use were used to determine the correlation between lipoprotein subclasses and coronary endothelial dysfunction. Epicardial endothelial dysfunction was significantly correlated with total (p=0.03) and small LDLp (LDL particles) (p<0.01) and inversely correlated with total and large HDLp (high density lipoprotein particles) (p<0.01). CONCLUSIONS: Epicardial, but not microvascular, coronary endothelial dysfunction was associated directly with LDL particles and inversely with HDL particles, suggesting location-dependent impact of lipoprotein particles on the coronary circulation

Response of macroinvertebrates to experimental nutrient and fish additions in European localities at different latitudes

The sedimentary and plant-associated macroinvertebrate communities have been examined in a series of mesocosm experiments carried out in five locations, in Finland, Sweden, UK, Northwest Spain and Mediterranean Spain (the International Mesocosm Experiment). The experiments were designed to investigate how shallow lake ecosystems respond to the addition of nutrients and planktivorous fish along a climate gradient. Nutrient additions increased sedimentary benthos densities and total biomass to a plateau at intermediate levels of nutrients. The effect was greater in the warmest localities. Beyond the saturation point a further increase in nutrient loading had different effects depending on locality. Only in the southernmost site (in Mediterranean Spain), did it induce a shift to a turbid phase and macrophyte disappearance. Thus there are breakpoints in the relationship with nutrients, leading to alternative sediment communities subsequent to the water alternative states. Presence of fish increased the densities and biomass of sediment macroinvertebrates, more clearly in the more southern locality, where the fish Gambusia did not prey in the bottom animals. A stabilising role of macrophytes was observed by cutting them, samples taken three weeks after cutting indicated a higher effect of fish on the sediment community when macrophytes were not present. Nutrients tended also to increase densities of plant associated macroinvertebrates at low levels with a flattening of response at intermediate levels and a decline at high levels. At high fish level the numbers and biomass of plant-associated macroinvertebrates were lowered in all cases, because of increased predation. However fish addition at low level in most cases had the opposite effect, because their predation pressure was not high and regeneration of nutrients through their activities might increase food availability and control invertebrate predators. Fish influenced especially community composition, because they prey selectively. Addition of fish at low densities increased macroinvertebrate diversity. A diel migratory cycle was observed in plant-associated macroinvertebrates, but only in the southerly locations. Latitudinal influences on the structure of the macroinvertebrate communities occur but that they operate indirectly. There was a prominent effect of nutrients on these communities so that starting trophy is very important. Fish effects are layered on these and depend to a large extent on the particular range of diet of the species involved.Se ha estudiado la comunidad de macroinvertebrados bentónicos del sedimento así como la de los asociados a los macrófitos en una serie de mesocosmos concebidos para un experimento múltiple simultáneo llevado a cabo en cinco localidades europeas: Finlandia, Suecia, Reino Unido (UK), Noroeste de España y Levante Español. El experimento fue diseñado para investigar como responden los ecosistemas de lagos someros a los aportes de nutrientes y a la abundancia de peces planctívoros a lo largo de un gradiente climático. La adición de nutrientes incrementó la densidad del bentos del sedimento y su biomasa hasta estabilizarse a niveles intermedios de nutrientes, siendo este efecto mayor en las latitudes más bajas y cálidas. Más allá del punto de saturación, un aumento adicional de la carga de nutrientes tuvo efectos diferentes dependiendo de la localidad y solo en el lugar de menor latitud, (en el Levante Español) indujo el cambio hacia una fase turbia, con desaparición de los macrófitos, esto es, hay puntos de ruptura relacionados con la carga de nutrientes que conducen a comunidades alternativas en el sedimento, subsiguientes a los mencionados estados alternativos en las aguas someras. La presencia de peces aumentó la densidad y biomasa de los macroinvertebrados del sedimento, ello también más claramente en la localidad situada más al Sur, ya que el pez planctívoro ensayado (Gambusia) por lo general no depreda en los animales del sedimento. Se observó también el papel estabilizador de los macrófitos comparando las muestras tomadas antes y después de que estos fueran cortados, por ejemplo se observó un efecto mayor de los peces en las comunidades del sedimento cuando los macrófitos no estaban presentes. Los nutrientes tendieron también a incrementar las densidades de los macroinvertebrados asociados a los macrófitos cuando se aportan a bajos niveles, con una estabilización de la respuesta para los valores intermedios y un declive para los valores más altos. Cuando se adicionaron densidades altas de peces, el número y biomasa de los macroinvertebrados asociados a los macrófitos descendieron en todos los casos, como resultado del aumento de la depredación. Sin embargo, las adiciones a baja densidad de peces, en la mayoría de los casos, tuvo el efecto contrario, ya que su presión de depredación no es alta y pueden contribuir a la regeneración de los nutrientes a través de la vía detrítica que incrementa la disponibilidad de alimento, así como al control de otros invertebrados depredadores. La presencia de peces influenció especialmente la composición de la comunidad, ya que su depredación es selectiva. La adición de peces en bajas densidades aumentó la diversidad de los macroinvertebrados. También fue observado que el ciclo diario de migración en los macroinvertebrados asociados a los macrófitos, es más importante en las localidades ubicadas en las latitudes de más al Sur. Las influencias latitudinales en la estructura de las comunidades de macroinvertebrados, si bien sucede, opera indirectamente. Hay un efecto dominante de los nutrientes en estas comunidades, y el nivel trófico de las condiciones iniciales es muy importante. Los efectos de los peces se subordinan al de los nutrientes y dependen en gran medida de la dieta de las especies involucradas

The Safety and Efficacy of Live Viral Vaccines in Patients With Cartilage-Hair Hypoplasia

Background: Live viral vaccines are generally contraindicated in patients with combined immunodeficiency including cartilage-hair hypoplasia (CHH); however, they may be tolerated in milder syndromes. We evaluated the safety and efficacy of live viral vaccines in patients with CHH.Methods: We analyzed hospital and immunization records of 104 patients with CHH and measured serum antibodies to measles, mumps, rubella, and varicella zoster virus (VZV) in all patients who agreed to blood sampling (n= 50). We conducted a clinical trial (identifier: NCT02383797) of live VZV vaccine on five subjects with CHH who lacked varicella history, had no clinical symptoms of immunodeficiency, and were seronegative for VZV; humoral and cellular immunologic responses were assessed post-immunization.Results: A large proportion of patients have been immunized with live viral vaccines, including measles-mumps-rubella (MMR) (n= 40, 38%) and VZV (n= 10, 10%) vaccines, with no serious adverse events. Of the 50 patients tested for antibodies, previous immunization has been documented with MMR (n= 22), rubella (n= 2) and measles (n= 1) vaccines. Patients with CHH demonstrated seropositivity rates of 96%/75%/91% to measles, mumps and rubella, respectively, measured at a medium of 24 years post-immunization. Clinical trial participants developed humoral and cellular responses to VZV vaccine. One trial participant developed post-immunization rash and knee swelling, both resolved without treatment.Conclusion: No serious adverse events have been recorded after immunization with live viral vaccines in Finnish patients with CHH. Patients generate humoral and cellular immune response to live viral vaccines. Immunization with live vaccines may be considered in selected CHH patients with no or clinically mild immunodeficiency

Immunopathological signatures in multisystem inflammatory syndrome in children and pediatric COVID-19

: Pediatric Coronavirus Disease 2019 (pCOVID-19) is rarely severe; however, a minority of children infected with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) might develop multisystem inflammatory syndrome in children (MIS-C), with substantial morbidity. In this longitudinal multi-institutional study, we applied multi-omics (analysis of soluble biomarkers, proteomics, single-cell gene expression and immune repertoire analysis) to profile children with COVID-19 (n = 110) and MIS-C (n = 76), along with pediatric healthy controls (pHCs; n = 76). pCOVID-19 was characterized by robust type I interferon (IFN) responses, whereas prominent type II IFN-dependent and NF-κB-dependent signatures, matrisome activation and increased levels of circulating spike protein were detected in MIS-C, with no correlation with SARS-CoV-2 PCR status around the time of admission. Transient expansion of TRBV11-2 T cell clonotypes in MIS-C was associated with signatures of inflammation and T cell activation. The association of MIS-C with the combination of HLA A*02, B*35 and C*04 alleles suggests genetic susceptibility. MIS-C B cells showed higher mutation load than pCOVID-19 and pHC. These results identify distinct immunopathological signatures in pCOVID-19 and MIS-C that might help better define the pathophysiology of these disorders and guide therapy

Interactions between Predation and Resources Shape Zooplankton Population Dynamics

Identifying the relative importance of predation and resources in population dynamics has a long tradition in ecology, while interactions between them have been studied less intensively. In order to disentangle the effects of predation by juvenile fish, algal resource availability and their interactive effects on zooplankton population dynamics, we conducted an enclosure experiment where zooplankton were exposed to a gradient of predation of roach (Rutilus rutilus) at different algal concentrations. We show that zooplankton populations collapse under high predation pressure irrespective of resource availability, confirming that juvenile fish are able to severely reduce zooplankton prey when occurring in high densities. At lower predation pressure, however, the effect of predation depended on algal resource availability since high algal resource supply buffered against predation. Hence, we suggest that interactions between mass-hatching of fish, and the strong fluctuations in algal resources in spring have the potential to regulate zooplankton population dynamics. In a broader perspective, increasing spring temperatures due to global warming will most likely affect the timing of these processes and have consequences for the spring and summer zooplankton dynamics

Pulmonary Follow-Up Imaging in Cartilage-Hair Hypoplasia : a Prospective Cohort Study

Cartilage-hair hypoplasia is a syndromic immunodeficiency with short stature, chondrodysplasia, and variable degree of immune dysfunction. Patients with cartilage-hair hypoplasia are prone to recurrent respiratory tract infections, and the prevalence of bronchiectasis ranges from 29 to 52%. Pulmonary complications contribute significantly to the mortality; therefore, regular lung imaging is essential. However, the optimal schedule for repeated lung imaging remains unestablished. We determined the rate and correlates of progression of structural lung changes in a prospectively followed cohort of 16 patients with cartilage-hair hypoplasia. We analyzed clinical, laboratory, and pulmonary functional testing data and performed lung magnetic resonance imaging at a median interval of 6.8 years since previous imaging. Imaging findings remained identical or improved due to disappearance of inflammatory changes in all evaluated patients. Patients with subtle signs of bronchiectasis on imaging tended to have low immunoglobulin M levels, as well as suffered from pneumonia during the follow-up. In conclusion, our results suggest slow if any development of bronchiectasis in selected subjects with cartilage-hair hypoplasia.Peer reviewe