Anatomical Laser Microdissection of the Ileum Reveals mtDNA Depletion Recovery in A Mitochondrial Neuro-Gastrointestinal Encephalomyopathy (MNGIE) Patient Receiving Liver Transplant

mitochondrial neuro-gastrointestinal encephalomyopathy (MNGIE) is a rare genetic disorder characterized by thymidine phosphorylase (TP) enzyme defect. The absence of TP activity induces the imbalance of mitochondrial nucleotide pool, leading to impaired mitochondrial DNA (mtDNA) replication and depletion. Since mtDNA is required to ensure oxidative phosphorylation, metabolically active tissues may not achieve sufficient energy production. The only effective life-saving approach in MNGIE has been the permanent replacement of TP via allogeneic hematopoietic stem cell or liver transplantation. However, the follow-up of transplanted patients showed that gut tissue changes do not revert and fatal complications, such as massive gastrointestinal bleeding, can occur. The purpose of this study was to clarify whether the reintroduction of TP after transplant can recover mtDNA copy number in a normal range. Using laser capture microdissection and droplet-digital-PCR, we assessed the mtDNA copy number in each layer of full-thickness ileal samples of a naive MNGIE cohort vs. controls and in a patient pre- and post-TP replacement. The treatment led to a significant recovery of gut tissue mtDNA amount, thus showing its efficacy. Our results indicate that a timely TP replacement is needed to maximize therapeutic success before irreversible degenerative tissue changes occur in MNGIE.     

Assessing the landscape context and conversion risk of protected areas using satellite data products

Since the establishment of the first national park (Yellowstone National Park in 1872) and the first wildlife refuge (Pelican Island in 1903), dramatic changes have occurred in both ecological and cultural landscapes across the U.S. The ability of these protected areas to maintain current levels of biodiversity depend, at least in part, on the integrity of the surrounding landscape. Our objective was to quantify and compare the extent and pattern of natural land cover, risk of conversion, and relationships with demographic and economic variables in counties near National Park Service units and U.S. Fish and Wildlife Service refuges with those counties distant from either type of protected area in the coterminous United States. Our results indicate that landscapes in counties within 10 km of both parks and refuges and those within 10 km of just parks were more natural, more intact, and more protected than those in counties within 10 km of just refuges and counties greater than 10 km from either protected area system. However, they also had greater human population density and change in population, indicating potential conversion risk since the percent of landscape protected averaged < 5% in both groups and human population dynamics are primary drivers of change in many landscapes. Conversion outweighed protection by at least two times (Conservation Risk Index > 2) in 76% of counties near both parks and refuges, 81% of counties near just parks, 91% of counties near just refuges, and 93% of distant counties. Thirteen percent of counties in the coterminous U.S. had moderate to high amounts of natural land cover (> 60%), low protection (< 20%), and the greatest change in population (> 20%). Although these areas are not the most critically endangered, they represent the greatest conservation opportunity, need, and urgency. Our approach is based on national level metrics that are simple, general, informative, and can be understood by broad audiences and by policy makers and managers to assess the health of lands surrounding parks and refuges. Regular monitoring of these metrics with satellite data products in counties surrounding protected areas provides a consistent, national level assessment of management opportunities and potentially adverse changes on adjacent lands.     

Competitive snoopy caching

In a snoopy cache multiprocessor system, each processor has a cache in which it stores blocks of data. Each cache is connected to a bus used to communicate with the other caches and with main memory. Each cache monitors the activity on the bus and in its own processor and decides which blocks of data to keep and which to discard. For several of the proposed architectures for snoopy caching systems, we present new on-line algorithms to be used by the caches to decide which blocks to retain and which to drop in order to minimize communication over the bus. We prove that, for any sequence of operations, our algorithms' communication costs are within a constant factor of the minimum required for that sequence; for some of our algorithms we prove that no on-line algorithm has this property with a smaller constant.A preliminary and condensed version of this paper appeared in theProceedings of the 27th Annual Symposium on the Foundations of Computer Science, IEEE, 1986.This author received support from an IBM doctoral fellowship, and did part of this work while a research student associate at IBM Almaden Research Center.     

Incremental Branching Programs

We propose a new model of restricted branching programs specific to solving GEN problems, which we call incremental branching programs. We show that syntactic incremental branching programs capture previously studied models of computation for the problem GEN, namely marking machines (Cook, S.A. in J. Comput. Syst. Sci. 9(3):308–316, 1974) and Poon’s extension (Proc. of the 34th IEEE Symp. on the Foundations of Computer Science, pp. 218–227, 1993) of jumping automata on graphs (Cook, S.A., Rackoff, C.W. in SIAM J. Comput. 9:636–652, 1980). We then prove exponential size lower bounds for our syntactic incremental model, and for some other variants of branching program computation for GEN. We further show that nondeterministic syntactic incremental branching programs are provably stronger than their deterministic counterpart when solving a natural NL-complete GEN sub-problem. It remains open if syntactic incremental branching programs are as powerful as unrestricted branching programs for GEN problems. A preliminary version of this paper appears as (Gál, A., Koucky, M., McKenzie, P., Incremental branching programs, in Proc. of the 2006 Computer Science in Russia Conference CSR06. Lecture Notes in Computer Science, vol. 3967, pp. 178–190, 2006). A. Gal supported in part by NSF Grant CCF-0430695 and an Alfred P. Sloan Research Fellowship. M. Koucky did part of this work while being a postdoctoral fellow at McGill University, Canada and at CWI, Amsterdam, Netherlands. Supported in part by NWO vici project 2004–2009, project No. 1M0021620808 of MŠMT ČR, grants 201/07/P276, 201/05/0124 of GA ČR, and Institutional Research Plan No. AV0Z10190503. P. McKenzie supported by the NSERC of Canada and the (Québec) FQRNT.     

A Dynamic Network Oligopoly Model with Transportation Costs,Product Differentiation,and Quality Competition

This paper develops a new dynamic model of Cournot–Nash oligopolistic competition that includes production and transportation costs, product differentiation, and quality levels in a network framework. The production costs capture the total quality cost, which, in turn, can also represent the R&D cost. We first present the equilibrium version and derive alternative variational inequality formulations. We then construct the projected dynamical systems model, which provides a continuous-time evolution of the firms’ product shipments and product quality levels, and whose set of stationary points coincides with the set of solutions to the variational inequality problem. We establish stability analysis results using a monotonicity approach and construct a discrete-time version of the continuous-time adjustment process, which yields an algorithm, with closed form expressions at each iteration. The algorithm is then utilized to compute solutions to several numerical examples. The framework can serve as the foundation for the modeling and analysis of competition among firms in industries ranging from food to pharmaceuticals to durable goods and high tech products, as well as Internet services, where quality and product differentiation are seminal.     

STUDIES ON YEAST ESTERASE

An esterase (carboxylic acid hydrolase EC 3 1.1.1.) has been prepared from Saccharomyces cerevisiae. This esterase is active at pH 4.4 and, though it is unstable in solution, it can be maintained in an active form either by lyophilization or by coupling to an affinity gel. At pH 4.4, yeast esterase can hydrolyse between 20 and 40% of the esters commonly present in beer, and it is capable of synthesizing ethyl acetate from ethanol and acetic acid in a simple buffered solution without the provision of a co-factor. Different yeast strains yield different amounts of esterase, and there appears to be a positive correlation between the ability of the yeast to form esterase and the level of esters present in fermentations accomplished by that yeast.     

Critical Review on Fatty Acid-Based Food and Nutraceuticals as Supporting Therapy in Cancer

Fatty acids have an important place in both biological and nutritional contexts and, from a clinical point of view, they have known consequences for diseases’ onset and development, including cancer. The use of fatty acid-based food and nutraceuticals to support cancer therapy is a multidisciplinary subject, involving molecular and clinical research. Knowledge regarding polyunsaturated fatty acids essentiality/oxidizability and the role of lipogenesis-desaturase pathways for cell growth, as well as oxidative reactivity in cancer cells, are discussed, since they can drive the choice of fatty acids using their multiple roles to support antitumoral drug activity. The central role of membrane fatty acid composition is highlighted for the application of membrane lipid therapy. As fatty acids are also known as biomarkers of cancer onset and progression, the personalization of the fatty acid-based therapy is also possible, taking into account other important factors such as formulation, bioavailability and the distribution of the supplementation. A holistic approach emerges combining nutra- and pharma-strategies in an appropriate manner, to develop further knowledge and applications in cancer therapy.     

Progression and Dissemination of Pulmonary Mycobacterium Avium Infection in a Susceptible Immunocompetent Mouse Model

Pulmonary infections caused by the group of nontuberculosis mycobacteria (NTM), Mycobacterium avium complex (MAC), are a growing public health concern with incidence and mortality steadily increasing globally. Granulomatous inflammation is the hallmark of MAC lung infection, yet reliable correlates of disease progression, susceptibility, and resolution are poorly defined. Unlike widely used inbred mouse strains, mice that carry the mutant allele at the genetic locus sst1 develop human-like pulmonary tuberculosis featuring well-organized caseating granulomas. We characterized pulmonary temporospatial outcomes of intranasal and left intrabronchial M. avium spp. hominissuis (M.av) induced pneumonia in B6.Sst1S mice, which carries the sst1 mutant allele. We utilized traditional semi-quantitative histomorphological evaluation, in combination with fluorescent multiplex immunohistochemistry (fmIHC), whole slide imaging, and quantitative digital image analysis. Followingintrabronchiolar infection with the laboratory M.av strain 101, the B6.Sst1S pulmonary lesions progressed 12–16 weeks post infection (wpi), with plateauing and/or resolving disease by 21 wpi. Caseating granulomas were not observed during the study. Disease progression from 12–16 wpi was associated with increased acid-fast bacilli, area of secondary granulomatous pneumonia lesions, and Arg1+ and double positive iNOS+/Arg1+ macrophages. Compared to B6 WT, at 16 wpi, B6.Sst1S lungs exhibited an increased area of acid-fast bacilli, larger secondary lesions with greater Arg1+ and double positive iNOS+/Arg1+ macrophages, and reduced T cell density. This morphomolecular analysis of histologic correlates of disease progression in B6.Sst1S could serve as a platform for assessment of medical countermeasures against NTM infection.     

Mitochondrial Dysfunction in Spinocerebellar Ataxia Type 3 Is Linked to VDAC1 Deubiquitination

Dysfunctional mitochondria are linked to several neurodegenerative diseases. Metabolic defects, a symptom which can result from dysfunctional mitochondria, are also present in spinocerebellar ataxia type 3 (SCA3), also known as Machado–Joseph disease, the most frequent, dominantly inherited neurodegenerative ataxia worldwide. Mitochondrial dysfunction has been reported for several neurodegenerative disorders and ataxin-3 is known to deubiquitinylate parkin, a key protein required for canonical mitophagy. In this study, we analyzed mitochondrial function and mitophagy in a patient-derived SCA3 cell model. Human fibroblast lines isolated from SCA3 patients were immortalized and characterized. SCA3 patient fibroblasts revealed circular, ring-shaped mitochondria and featured reduced OXPHOS complexes, ATP production and cell viability. We show that wildtype ataxin-3 deubiquitinates VDAC1 (voltage-dependent anion channel 1), a member of the mitochondrial permeability transition pore and a parkin substrate. In SCA3 patients, VDAC1 deubiquitination and parkin recruitment to the depolarized mitochondria is inhibited. Increased p62-linked mitophagy, autophagosome formation and autophagy is observed under disease conditions, which is in line with mitochondrial fission. SCA3 fibroblast lines demonstrated a mitochondrial phenotype and dysregulation of parkin-VDAC1-mediated mitophagy, thereby promoting mitochondrial quality control via alternative pathways.