Strictosidine, the common precursor for monoterpenoid indole alkaloids with 3 α and 3 β configuration

Recently we reported that strictosidine (l> is the key intermediate in the formation of the three classes (ASDidOf3DeIm5, Iboga, Corynanthe) of monoterpenoid indole alkaloids In Catharanthus roseus and a variety of other plant species in cell culture using in vivo and in vitro techniques"2. These results were independently confirmed In Manchester 394 and subsequently also Scott et a1.5 were able to confirm the precursor role of (1) using Catharanthus material. All these results are in accord with reports on the biosynthesis of an alkaloid of taxonomically distant origin, camptothecln, for which strictosidine la&am6 was previously found to be a precursor, and recently7 also (1). The key intermediate in the biosynthesis of the majority of mono-terpenoid alkaloids is therefore (1) with 3 a (2) configuration, rather than vincoside (2) with 3 13 (lX> configuration as had previously been assumeda. However, a generalization of this precursor function of (1

Rejuvenation by cell reprogramming: A new horizon in gerontology

The discovery of animal cloning and subsequent development of cell reprogramming technology were quantum leaps as they led to the achievement of rejuvenation by cell reprogramming and the emerging view that aging is a reversible epigenetic process. Here, we will first summarize the experimental achievements over the last 7 years in cell and animal rejuvenation. Then, a comparison will be made between the principles of the cumulative DNA damage theory of aging and the basic facts underlying the epigenetic model of aging, including Horvath's epigenetic clock. The third part will apply both models to two natural processes, namely, the setting of the aging clock in the mammalian zygote and the changes in the aging clock along successive generations in mammals. The first study demonstrating that skin fibroblasts from healthy centenarians can be rejuvenated by cell reprogramming was published in 2011 and will be discussed in some detail. Other cell rejuvenation studies in old humans and rodents published afterwards will be very briefly mentioned. The only in vivo study reporting that a number of organs of old progeric mice can be rejuvenated by cyclic partial reprogramming will also be described in some detail. The cumulative DNA damage theory of aging postulates that as an animal ages, toxic reactive oxygen species generated as byproducts of the mitochondria during respiration induce a random and progressive damage in genes thus leading cells to a progressive functional decline. The epigenetic model of aging postulates that there are epigenetic marks of aging that increase with age, leading to a progressive derepression of DNA which in turn causes deregulated expression of genes that disrupt cell function. The cumulative DNA damage model of aging fails to explain the resetting of the aging clock at the time of conception as well as the continued vitality of species as millenia go by. In contrast, the epigenetic model of aging straightforwardly explains both biologic phenomena. A plausible initial application of rejuvenation in vivo would be preventing adult individuals from aging thus eliminating a major risk factor for end of life pathologies. Further, it may allow the gradual achievement of whole body rejuvenation.Fil: Goya, Rodolfo Gustavo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias Médicas. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; ArgentinaFil: Lehmann, Marianne. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias Médicas. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; ArgentinaFil: Chiavellini, Priscila. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias Médicas. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; ArgentinaFil: Canatelli Mallat, Martina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias Médicas. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; ArgentinaFil: Hereñú, Claudia Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Farmacología Experimental de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Instituto de Farmacología Experimental de Córdoba; ArgentinaFil: Brown, Oscar Alfredo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias Médicas. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; Argentin

Metformin treatment for diabetes mellitus correlates with progression and survival in colorectal carcinoma

BACKGROUND: Diabetes mellitus is unfavorably associated with cancer risk. The purpose of this multi-disciplinary project was to evaluate a possible association of diabetes mellitus and other comorbidities and their treatment with progression of colorectal cancer. PATIENTS AND METHODS: We investigated the correlation between pathological characteristics and clinical course, including comorbidities in 1004 Czech patients diagnosed and surgically treated for colorectal adenocarcinoma (CRC) between 1999 and 2016. RESULTS: In our data set, CRC patients treated with metformin due to coexisting diabetes mellitus type 2 (T2DM) developed fewer distant metastases which clinically correlates with slower CRC progression. Survival in metformin subgroup was longer, particularly in men with CRC. Osteoporosis may be a negative factor of survival in CRC patients. CONCLUSIONS: Our findings also indicate that aging, higher tumor grade and TNM stage, coexistence of selected endocrine disorders, and metabolic abnormalities may change the tumor microenvironment and impact survival in colorectal cancer, although mechanism of these observations yet to be explained. Patients with diabetes mellitus type 2 treated with metformin may represent the altered microenvironment with specifically tuned metabolic molecular responses and with various epigenetic characteristics. More awareness and increased understanding of the mechanisms underlying the positive effect of metformin on patients' survival could offer insight into new treatment methods and permit more individualized treatment plans.Web of Science13239238

Characterisation and process verification studies in a miniature bioreactor used as a predictive tool to scale-up an industrial process

There is continuous pressure on the pharmaceutical industry to acquire a thorough understanding of their product and process, using a quality by design (QbD) approach, in order to speed up the timescales for approval of a therapy to market. One way to achieve this is to conduct detailed characterization studies using a miniature model, such as a bioreactor or a single-use micro-well plate system, in order to identify the key parameters which have an impact on the scale-up of a bioprocess. The design and development of such a representative miniature model, using integrated sensors for monitoring of process conditions on-line, is vital for scaling purposes and allows for parallelization and automation whereby multiple parameters can be tested in a high-throughput fashion. This concept has been applied to a downstream process for the recovery of periplasmic Fab’ from E.coli cells, a therapeutic recombinant protein used to treat autoimmune diseases. The aim of the research is to address the key challenges of scaling by characterizing a 20mL small scale model in terms of mixing performance and fluid dynamics and identifying which factors, such as shear stress, fluid velocity and spatial distribution of cells have the most impact on scaling. In the study, novel, advanced techniques, such as the dual pH indicator system for mixing time (DISMT) and particle image velocimetry (PIV), were used to characterise the fluid dynamics in the small scale bioreactor. The bioreactor was fitted with a miniature temperature and pH probe to observe conditions, and process verification studies were conducted in order to compare performance at different scales for a variety of operating conditions. The results show that the small scale bioreactor was able to successfully mimic an industrially relevant heat extraction process up to 10,000 fold scale with good accuracy on the basis of constant volumetric power input. Assays were used to analyse the quantity and quality of the product and impurities, and scanning electron microscopy was used to examine the morphology of cells at different stages of the process. The study was further extended to 24 deep square micro-well plates and initial data suggests that the shaken system may also be used to scale-down 10,000 fold, and therefore indicates the feasibility and applicability of these single-use plates as a predictive tool for scaling up. The poster will discuss how fluid dynamic studies may be used to understand and improve scale-up and will additionally address alternative methods which may be used to minimise the variation observed in the feed material and hence its impact on subsequent unit operations

Measuring what matters in healthcare: A practical guide to psychometric principles and instrument development

The provision of quality healthcare relies on scales and measures with robust evidence of their psychometric properties. Using measurement instruments with poor reliability, validity, or feasibility, or those that are not appropriate for the target diagnostic group or construct/dimension under consideration, may be unfavorable for patients, unproductive, and hinder empirical advancement. Resources from the COnsensus-based Standards for the selection of health status Measurement INstruments (COSMIN) group can assist in identifying and developing psychometrically sound measures. The COSMIN initiative is the only international, research-based practice taxonomy and methodological guidelines for measurement in healthcare. This manuscript aims to provide an accessible introduction to theories, principles and practices of psychometrics, instrument properties, and scale development, with applied examples from the COSMIN recommendations. It describes why measurement in healthcare is critical to good practice, explains the concepts of the latent variable and hypothetical construct and their importance in healthcare assessments, explores issues of flawed measurement and briefly explains key theories relevant to psychometrics. The paper also outlines a ten-step process to develop and validate a new measurement instrument, with examples drawn from a recently developed visuoperceptual measure for analysis of disordered swallowing to demonstrate key concepts and provides a guide for understanding properties of and terminology related to measurement instruments. This manuscript serves as a resource for healthcare clinicians, educators, and researchers who seek to develop and validate new measurement instruments or improve the properties of existing ones. It highlights the importance of using psychometrically sound measurement instruments to ensure high-quality healthcare assessments

Waiting for shelter: Perspectives on a homeless shelter’s procedures

Research on homeless shelter implementation is limited. Some shelters have lengthy waitlists, which raises important questions about implications of waitlists for individuals with immediate shelter needs. This study used qualitative methods to understand the experiences of shelter seekers who were on a shelter waitlist (N = 59), including individuals who entered the shelter from the waitlist, and those removed from the shelter waitlist for procedural reasons. The average waitlist time was nearly 3 weeks, and 22.0% stayed at least one night on the street or another public place while on the waitlist. Responses to open‐ended questions regarding barriers and effectiveness of the shelter referral procedures revealed 4 themes: procedural challenges, procedural benefits, benefits of the temporary stay, and communication challenges. Further research is needed to inform shelter implementation on a larger scale in accordance with current community‐wide efforts to coordinate shelter services.Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/138221/1/jcop21896_am.pdfhttps://deepblue.lib.umich.edu/bitstream/2027.42/138221/2/jcop21896.pd

ALS-associated missense and nonsense TBK1 mutations can both cause loss of kinase function

Mutations in TANK binding kinase 1 (TBK1) have been linked to amyotrophic lateral sclerosis. Some TBK1 variants are nonsense and are predicted to cause disease through haploinsufficiency; however, many other mutations are missense with unknown functional effects. We exome sequenced 699 familial amyotrophic lateral sclerosis patients and identified 16 TBK1 novel or extremely rare protein-changing variants. We characterized a subset of these: p.G217R, p.R357X, and p.C471Y. Here, we show that the p.R357X and p.G217R both abolish the ability of TBK1 to phosphorylate 2 of its kinase targets, IRF3 and optineurin, and to undergo phosphorylation. They both inhibit binding to optineurin and the p.G217R, within the TBK1 kinase domain, reduces homodimerization, essential for TBK1 activation and function. Finally, we show that the proportion of TBK1 that is active (phosphorylated) is reduced in 5 lymphoblastoid cell lines derived from patients harboring heterozygous missense or in-frame deletion TBK1 mutations. We conclude that missense mutations in functional domains of TBK1 impair the binding and phosphorylation of its normal targets, implicating a common loss of function mechanism, analogous to truncation mutations

Global impacts of energy demand on the freshwater resources of nations

The growing geographic disconnect between consumption of goods, the extraction and processing of resources, and the environmental impacts associated with production activities makes it crucial to factor global trade into sustainability assessments. Using an empirically validated environmentally extended global trade model, we examine the relationship between two key resources underpinning economies and human well-being—energy and freshwater. A comparison of three energy sectors (petroleum, gas, and electricity) reveals that freshwater consumption associated with gas and electricity production is largely confined within the territorial boundaries where demand originates. This finding contrasts with petroleum, which exhibits a varying ratio of territorial to international freshwater consumption, depending on the origin of demand. For example, although the United States and China have similar demand associated with the petroleum sector, international freshwater consumption is three times higher for the former than the latter. Based on mapping patterns of freshwater consumption associated with energy sectors at subnational scales, our analysis also reveals concordance between pressure on freshwater resources associated with energy production and freshwater scarcity in a number of river basins globally. These energy-driven pressures on freshwater resources in areas distant from the origin of energy demand complicate the design of policy to ensure security of fresh water and energy supply. Although much of the debate around energy is focused on greenhouse gas emissions, our findings highlight the need to consider the full range of consequences of energy production when designing policy

Fracture of a biopolymer gel as a viscoplastic disentanglement process

We present an extensive experimental study of mode-I, steady, slow crack dynamics in gelatin gels. Taking advantage of the sensitivity of the elastic stiffness to gel composition and history we confirm and extend the model for fracture of physical hydrogels which we proposed in a previous paper (Nature Materials, doi:10.1038/nmat1666 (2006)), which attributes decohesion to the viscoplastic pull-out of the network-constituting chains. So, we propose that, in contrast with chemically cross-linked ones, reversible gels fracture without chain scission