Molecular recognition of diketide substrates by a beta-ketoacyl-acyl carrier protein synthase domain within a bimodular polyketide synthase

BACKGROUND: Modular polyketide synthases (PKSs) are large multifunctional proteins that catalyze the biosynthesis of structurally complex bioactive products. The modular organization of PKSs has allowed the application of a combinatorial approach to the synthesis of novel polyketides via the manipulation of these biocatalysts at the genetic level. The inherent specificity of PKSs for their natural substrates, however, may place limits on the spectrum of molecular diversity that can be achieved in polyketide products. With the aim of further understanding PKS specificity, as a route to exploiting PKSs in combinatorial synthesis, we chose to examine the substrate specificity of a single intact domain within a bimodular PKS to investigate its capacity to utilize unnatural substrates. RESULTS: We used a blocked mutant of a bimodular PKS in which formation of the triketide product could occur only via uptake and processing of a synthetic diketide intermediate. By introducing systematic changes in the native diketide structure, by means of the synthesis of unnatural diketide analogs, we have shown that the ketosynthase domain of module 2 (KS2 domain) in 6-deoxyerythronolide B synthase (DEBS) tolerates a broad range of variations in substrate structure, but it strongly discriminates against some others. CONCLUSIONS: Defining the boundaries of substrate recognition within PKS domains is crucial to the rationally engineered biosynthesis of novel polyketide products, many of which could be prepared only with great difficulty, if at all, by direct chemical synthesis or semi-synthesis. Our results suggest that the KS2 domain of DEBS1 has a relatively relaxed specificity that can be exploited for the design and synthesis of medicinally important polyketide products.     

Socially Assistive Robots in Elderly Care: A Mixed-Method Systematic Literature Review

The world’s population is aging, and developed countries are engaged in developing a new aged-care paradigm to reduce spiraling healthcare costs. Assistive technologies like Socially Assistive Robots (SAR) are being considered as enablers to support the process of care giving or keep elderly at home longer. This article reports a mixed-method systematic review of SAR in elderly care and recognizes its impact on elderly well-being, integrating evidence from qualitative and quantitative studies. It follows the principles explained in Cochrane Handbook for Systematic Reviews of Interventions and classifies interventions, measures, and outcomes of field trials of SAR in elderly care. Eighty-six studies in 37 study groups have been included. The findings imply positive effects of SAR on elderly well-being. Ten significant recommendations are made to help avoid the current limitations of existing research and to improve future research and its applicability. This review revealed that SAR can potentially enhance elderly well-being and decrease the workload on caregivers. There is a need for rigorous research methodology, person-centered care, caregiver expectation model, multimodal interaction, multimodal data collection, and modeling of culturally diverse groups to facilitate acceptability of SAR.     

Survey of denoising,segmentation and classification of magnetic resonance imaging for prostate cancer

Multimedia Tools and Applications - Prostate cancer (PCa) has become the second most dreadful cancer in men after lung cancer. Traditional approaches used for treatment of PCa were manual, time...     

Characteristic study of N07718 superalloy surface prepared by cold spray

For better performance in erosive–corrosive, wear and other extreme environmental conditions surface properties of metals and alloys are usually altered by different techniques. Amongst the various surface coating techniques cold spraying is a relatively new and less reported method. Coating formation at energy lower than the melting stage of materials brings novelty to this process. In this study surface of UNS No. - N07718 alloy is altered by cold spraying chromium-carbide (nickel-chromium) layer. The microstructure properties of this cold sprayed surface are observed by various characterization techniques and the results are used to study mechanism of coating layer formation by cold spray process. This newly developed cold sprayed surface is found to be eligible for further testing in wear and erosive-corrosive environmental conditions.     

Proteome of human endometrium: Identification of differentially expressed proteins in proliferative and secretory phase endometrium

Purpose: To exploit the potential of proteomics to identify and study additional yet‐unidentified important proteins present in human endometrium. Experimental design: The proteome of human endometrium would be established using 2‐DE and MALDI and the data analyzed to identify differential protein expression in the proliferative and secretory phase of the menstrual cycle using PDQuest software and MALDI. Results: In the present work, 2‐DE of human endometrium protein led to the resolution of over 200 spots. Subsequent MALDI analysis of 215 spots allowed the identification of 194 proteins. A total of 57 out of the 215 spots were found to be differentially expressed, out of which 49 could be identified using MALDI. These differentially expressed proteins included structural proteins, molecular chaperones, signaling proteins, metabolic proteins, proteins related to immunity, RNA biogenesis, protein biosynthesis and others. The differential expressions of seven representative proteins in secretory and proliferative phase endometrium tissue were confirmed by immunoblot analysis. Conclusion and clinical relevance: This study establishes the 2‐D proteome of human endometrium represented by 194 identified protein spots. The present data provides an important clue towards determining the function of these proteins with respect to endometrium related diseases.     

Plasma lipids are affected similarly by dietary lauric or palmitic acid in gerbils and monkeys

To compare the relative impact of dietary lauric acid (12∶0) and palmitic acid (16∶0) on plasma lipids, two fat-sensitive species, Mongolian gerbils and cebus monkeys, were fed cholesterol-free, purified diets enriched with either 12∶0-rich or 16∶0-rich fats, while all other fatty acids were held constant by selective blending of up to five natural fats or oils. The two gerbil diets (40 en% from fat) allowed for an 8 en% exchange between 12∶0 and 16∶0, and the monkey diets (31 en% from fat) allowed for 6 en% exchange beteen these two fatty acids. Eight gerbils received the diets for eight weeks, and 12 cebus monkeys were fed each diet in a cross-over design for up to 22 wk. Both diets resulted in similar plasma cholesterol, triglyceride, and high density lipoprotein cholesterol concentrations within each species. Additionally, separation of cebus lipoproteins by discontinuous density-gradient ultracentrifugation failed to show any dietary differences in concentration or composition of the three major lipoprotein classes (d<1.019, 1.019–1.055, and 1.055–1.168 g/mL). Thus, in two species sensitive to manipulations in dietary fat while consuming cholesterol-free diets, 16∶0 was not hypercholesterolemic relative to 12∶0. Based on a paper presented at the PORIM International Palm Oil Congress (PIPOC) held in Kuala Lumpur, Malaysia, September 1993.     

Manual Therapy Facilitates Homeostatic Adaptation to Bone Microstructural Declines Induced by a Rat Model of Repetitive Forceful Task

The effectiveness of manual therapy in reducing the catabolic effects of performing repetitive intensive force tasks on bones has not been reported. We examined if manual therapy could reduce radial bone microstructural declines in adult female Sprague–Dawley rats performing a 12-week high-repetition and high-force task, with or without simultaneous manual therapy to forelimbs. Additional rats were provided 6 weeks of rest after task cessation, with or without manual therapy. The control rats were untreated or received manual therapy for 12 weeks. The untreated TASK rats showed increased catabolic indices in the radius (decreased trabecular bone volume and numbers, increased osteoclasts in these trabeculae, and mid-diaphyseal cortical bone thinning) and increased serum CTX-1, TNF-α, and muscle macrophages. In contrast, the TASK rats receiving manual therapy showed increased radial bone anabolism (increased trabecular bone volume and osteoblast numbers, decreased osteoclast numbers, and increased mid-diaphyseal total area and periosteal perimeter) and increased serum TNF-α and muscle macrophages. Rest, with or without manual therapy, improved the trabecular thickness and mid-diaphyseal cortical bone attributes but not the mineral density. Thus, preventive manual therapy reduced the net radial bone catabolism by increasing osteogenesis, while rest, with or without manual therapy, was less effective.     

Engineering the acyltransferase substrate specificity of assembly line polyketide synthases

Polyketide natural products act as a broad range of therapeutics, including antibiotics, immunosuppressants and anti-cancer agents. This therapeutic diversity stems from the structural diversity of these small molecules, many of which are produced in an assembly line manner by modular polyketide synthases. The acyltransferase (AT) domains of these megasynthases are responsible for selection and incorporation of simple monomeric building blocks, and are thus responsible for a large amount of the resulting polyketide structural diversity. The substrate specificity of these domains is often targeted for engineering in the generation of novel, therapeutically active natural products. This review outlines recent developments that can be used in the successful engineering of these domains, including AT sequence and structural data, mechanistic insights and the production of a diverse pool of extender units. It also provides an overview of previous AT domain engineering attempts, and concludes with proposed engineering approaches that take advantage of current knowledge. These approaches may lead to successful production of biologically active ‘unnatural’ natural products.