Caffeine, but not other phytochemicals, in mate tea (Ilex paraguariensis St. Hilaire) attenuates high-fat-high-sucrose-diet-driven lipogenesis and body fat accumulation

Authors followed the guidelines of the National Institutes of Health guide for the care and use of Laboratory animals (NIH Publications No.8023, revised 1978)The objective was to examine the effectiveness of mate tea (MT, Ilex paraguariensis St. Hilaire) and caffeine from mate tea (MC) on in vitro lipid accumulation and in vivo diet-driven-obesity. MC and decaffeinated mate (DM) were obtained using supercritical CO2 extraction and mainly composed of caffeine and caffeoylquinic acids, respectively. MC reduced lipid accumulation (41%) via downregulation of fatty acid synthase (Fasn) (39%) in 3T3-L1 adipocytes. Rats fed a high-fat-high-sucrose-diet and 0.1% of caffeine from MC, MT, or DM. MC attenuated weight gain (16%) and body fat accumulation (22%). MC reduced Fasn expression in both adipose tissue (66%) and liver (37%). MC diminished pyruvate kinase (PK, 59%) and microsomal triglyceride transfer protein (MTP, 50%) hepatic expression. In silico, neochlorogenic acid interacted with PK and MTP allosteric sites. FAS β‐ketoacyl reductase domain showed the highest affinity to 3,5-dicaffeoylquinic acid. Caffeine suppressed lipid accumulation and body weight gain, through the modulation of lipogenic gene expressio

Expansion and Harvesting of hMSC-TERT

The expansion of human mesenchymal stem cells as suspension culture by means of spinner flasks and microcarriers, compared to the cultivation in tissue culture flasks, offers the advantage of reducing the requirements of large incubator capacities as well as reducing the handling effort during cultivation and harvesting. Nonporous microcarriers are preferable when the cells need to be kept in viable condition for further applications like tissue engineering or cell therapy. In this study, the qualification of Biosilon, Cytodex 1, Cytodex 3, RapidCell and P102-L for expansion of hMSC-TERT with an associated harvesting process using either trypsin, accutase, collagenase or a trypsin-accutase mixture was investigated. A subsequent adipogenic differentiation of harvested hMSC-TERT was performed in order to observe possible negative effects on their (adipogenic) differentiation potential as a result of the cultivation and harvesting method. The cultivated cells showed an average growth rate of 0.52 d-1. The cells cultivated on Biosilon, RapidCell and P102-L were harvested succesfully achieving high cell yield and vitalities near 100%. This was not the case for cells on Cytodex 1 and Cytodex 3. The trypsin-accutase mix was most effective. After spinner expansion and harvesting the cells were successfully differentiated to adipocytes

Diffusion of Protease into Meat & Bone Meal for Solubility Improvement and Potential Inactivation of the BSE Prion

BACKGROUND: Government-imposed feed bans have created a need for new applications for meat & bone meal (MBM). Many potential new applications require MBM protein to be both soluble and free of infectious prion. Treatment with protease is generally effective in reducing insoluble, thermally-denatured proteins to soluble peptides. It has been reported in the literature that certain proteases, including Versazyme™, are able to degrade infectious prions in a system where the prions are readily accessible to proteolytic attack. Prions distributed within MBM, however, may conceivably be protected from proteases. METHODOLOGY/PRINCIPAL FINDINGS: The overall rate of proteolytic MBM digestion depends greatly on whether the protease can penetrate deep within individual particles, or if the protease can only act near the surface of the particle. This research examined the barriers to the diffusion of Versazyme™ into particles of MBM. Confocal microscopy demonstrated differences in the density distributions between the bone and the soft tissue particles of MBM. By tracking the diffusion of fluorescently labeled Versazyme™ through individual particles, it was found that bone particles show full Versazyme™ penetration within 30 minutes, while penetration of soft tissue particles can take up to four hours, depending on the particle's diameter. From the variety of normal proteins comprising MBM, a specific protein was chosen to serve as a prion surrogate based on characteristics including size, solubility, distribution and abundance. This surrogate was used to measure the effect of several factors on Versazyme™ diffusion. CONCLUSIONS/SIGNIFICANCE: Results showed that surrogate distributed in bone particles was more susceptible to degradation than that in soft tissue particles. Three factors controllable by unit operations in an industrial-scale process were also tested. It was found that removing the lipid content and hydrating MBM prior to incubation both significantly increased the rate of surrogate degradation. In a test of particle size, the smallest collected diameter range demonstrated the largest degradation of the prion surrogate, suggesting milling would be beneficial

An overview of animal prion diseases

Prion diseases are transmissible neurodegenerative conditions affecting human and a wide range of animal species. The pathogenesis of prion diseases is associated with the accumulation of aggregates of misfolded conformers of host-encoded cellular prion protein (PrPC). Animal prion diseases include scrapie of sheep and goats, bovine spongiform encephalopathy (BSE) or mad cow disease, transmissible mink encephalopathy, feline spongiform encephalopathy, exotic ungulate spongiform encephalopathy, chronic wasting disease of cervids and spongiform encephalopathy of primates. Although some cases of sporadic atypical scrapie and BSE have also been reported, animal prion diseases have basically occurred via the acquisition of infection from contaminated feed or via the exposure to contaminated environment. Scrapie and chronic wasting disease are naturally sustaining epidemics. The transmission of BSE to human has caused more than 200 cases of variant Cruetzfeldt-Jacob disease and has raised serious public health concerns. The present review discusses the epidemiology, clinical neuropathology, transmissibility and genetics of animal prion diseases

In vitro models for the study of osteoarthritis

AbstractOsteoarthritis (OA) is a prevalent disease of most mammalian species and is a significant cause of welfare and economic morbidity in affected individuals and populations. In vitro models of osteoarthritis are vital to advance research into the causes of the disease, and the subsequent design and testing of potential therapeutics. However, a plethora of in vitro models have been used by researchers but with no consensus on the most appropriate model. Models attempt to mimic factors and conditions which initiate OA, or dissect the pathways active in the disease. Underlying uncertainty as to the cause of OA and the different attributes of isolated cells and tissues used mean that similar models may produce differing results and can differ from the naturally occurring disease.This review article assesses a selection of the in vitro models currently used in OA research, and considers the merits of each. Particular focus is placed on the more prevalent cytokine stimulation and load-based models. A brief review of the mechanism of these models is given, with their relevance to the naturally occurring disease. Most in vitro models have used supraphysiological loads or cytokine concentrations (compared with the natural disease) in order to impart a timely response from the cells or tissue assessed. Whilst models inducing OA-like pathology with a single stimulus can answer important biological questions about the behaviour of cells and tissues, the development of combinatorial models encompassing different physiological and molecular aspects of the disease should more accurately reflect the pathogenesis of the naturally occurring disease

Prion protein gene sequence and chronic wasting disease susceptibility in white-tailed deer ( Odocoileus virginianus

The sequence of the prion protein gene (PRNP) affects susceptibility to spongiform encephalopathies, or prion diseases in many species. In white-tailed deer, both coding and non-coding single nucleotide polymorphisms have been identified in this gene that correlate to chronic wasting disease (CWD) susceptibility. Previous studies examined individual nucleotide or amino acid mutations; here we examine all nucleotide polymorphisms and their combined effects on CWD. A 626 bp region of PRNP was examined from 703 free-ranging white-tailed deer. Deer were sampled between 2002 and 2010 by hunter harvest or government culling in Illinois and Wisconsin. Fourteen variable nucleotide positions were identified (4 new and 10 previously reported). We identified 68 diplotypes comprised of 24 predicted haplotypes, with the most common diplotype occurring in 123 individuals. Diplotypes that were found exclusively among positive or negative animals were rare, each occurring in less than 1% of the deer studied. Only one haplotype (C, odds ratio 0.240) and 2 diplotypes (AC and BC, odds ratios of 0.161 and 0.108 respectively) has significant associations with CWD resistance. Each contains mutations (one synonymous nucleotide 555C/T and one nonsynonymous nucleotide 286G/A) at positions reported to be significantly associated with reduced CWD susceptibility. Results suggest that deer populations with higher frequencies of haplotype C or diplotypes AC and BC might have a reduced risk for CWD infection – while populations with lower frequencies may have higher risk for infection. Understanding the genetic basis of CWD has improved our ability to assess herd susceptibility and direct management efforts within CWD infected areas

The impact of maternal infection with chronic wasting disease on fetal characteristics in wild white-tailed deer (Odocoileus virginianus) in Illinois, USA

abstract: Chronic wasting disease (CWD) is a fatal neurodegenerative disease of cervids caused by a misfolded and infectious protein, called a prion. Infection with CWD is characterized by an asymptomatic period, followed by a decline that inevitably leads to death. The disease is transmitted mainly through direct contact between infected and uninfected animals, but vertical transmission has been documented in muntjac deer (Muntiacus reevesi), along with the presence of infectious prions in the reproductive tissues of elk (Cervus canadensis) and white-tailed deer (Odocoileus virginianus). Therefore, questions have been raised about the effects of CWD on the health and characteristics of the offspring gestated by infected animals, and about how vertical transmission may complicate disease management efforts and drive reduction of herd fitness. To begin finding quantitative answers to these questions, this study used multilevel Bayesian linear regression to model the influence of maternal infection with CWD, along with several other environmental and biological variables, on fetal weight, sex, and length using data collected on white-tailed deer as part of the CWD management program in Illinois for the time period between fiscal years 2003 and 2020. The regression analyses found no statistically significant relationship between maternal CWD status and fetal length or the fraction of males in a litter but did find a significant relationship between maternal infection and fetal weight. Fetuses of CWD positive deer weighed 1% less than fetuses from deer in which CWD was not detected. Given that 88% of the fetuses in this data were in the second trimester of gestation, the impact of CWD on fetuses through the entire course of gestation remains to be evaluated