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Microbial keratinases as potential therapeutic options for neurodegenerative diseases

Posted on February 10, 2022 By Clayton No Comments on Microbial keratinases as potential therapeutic options for neurodegenerative diseases
Background: Normal proteins, if misfolded, form insoluble aggregates or fibrils, resistant to proteolytic degradation. Such proteins, named amyloid proteins, share a common structural motif, called the cross β sheet structure.
The insoluble β-amyloid fibrils have been implicated in neurodegenerative diseases such as Alzheimer’s disease (AD). Prions have structural similarities with β-amyloid fibrils. As microbial keratinases can degrade prions, we investigated the possible capacity of microbial keratinases, purified in our lab, for degrading β- amyloid fibrils.
Method: Lysozyme is a model system to study protein misfolding. β-Amyloid fibrils of lysozyme were prepared by treating Hen Egg White Lysozyme (HEWL) with 8M Urea for 48 h at 55°C.
The fibril formation was confirmed by formation of a fibrous precipitate, immunoblotting with anti β-amyloid antibody, High Performance Liquid Chromatography (HPLC), and Congo Red Absorption spectroscopy. Using Q Sepharose Column Chromatography, two microbial keratinases, Ker1 and Ker2 were purified from an actinomycete strain, Amycolatopsis sp. MBRL 40, previously isolated in our laboratory (MBRL). β-amyloid fibril degrading activity of these keratinases were investigated both in solution or as reconstituted form on cationic/neutral liposomes.
Result: Both soluble and reconstituted Ker1 degraded β-amyloid fibrils as confirmed by loss of fibrous precipitate of β-amyloid fibrils, changes in the spectra as observed in HPLC and Congo Red Absorption spectroscopy and loss of signal when immunoblotted with anti β-amyloid antibody.
Details of the findings on degradation of β-amyloid degradation by keratinase (Ker1) of indigenous Amycolatopsis sp. MBRL 40 obtained in our laboratory and its potential as a therapeutic option for neurodegenerative diseases shall be presented in this oral paper.
Conclusion: Many drugs for treating Alzheimer’s disease (AD) are in various stages of clinical trials. These drugs either inhibit the activity of acetylcholinesterase or inhibit the formation of Aβ plaques and their downstream effects e.g. Ca++ channel antagonists, nonsteroidal anti-inflammatory drugs, antioxidants, iron chelators, hypolipidemic drugs etc. Microbial keratinases are novel molecules in this sense as they haven’t been investigated so far and we propose these enzymes as new potential therapeutic interventions for neurodegenerative diseases such as AD.
Background: In a search for innovative dementia care strategies, a new research field has been dedicated to design social robots for older adults with cognitive impairments and dementia in order to promote their quality of life through communication and social interactions.
Although the results are promising, the quality of studies on effectiveness of social robots in elderly adults is still low due to several methodological limitations (1). Therefore, we aimed to review the methodologies used so far for conducting studies with social robots in long-term and community care settings with people with dementia and elderly adults.
Method: Searches in PubMed, Web of Science, PsycInfo, and Cinahl yielded 33 studies included in the final review after applying inclusion and exclusion criteria. Reviews were conducted by two authors independently.
Result: Across 33 papers, 23 different social robots were examined on elderly adults and PwD, investigating acceptability, usability and effectiveness.
A variety of study designs such as RCTs, mixed methods, cross-sectional, cohort and quasi-experimental designs were employed. Individual and group-based interventions took place in private households, long-term care facilities and laboratories during single or multiple sessions Gentaur Laboratory Paper. Data were collected through observation, questionnaires, interviews and physiological tests.
Most revealed improvements were found in mood, engagement and participation. However, improvements in quality of life, depression and cognition were not significant.
Conclusion: Despite the promising results, due to the insufficient methodology, failures and ambitious findings remain within the many aspects of the enquiry. Appropriate experimental designs such as RCTs with large sample sizes and individual intervention sessions might be established for effectiveness studies. For acceptability/usability studies, a cohort design with long-term intervention would serve best. We also recommend a mixed-method of data collection for any of three effectiveness, acceptability and usability related outcomes. Multiple interaction sessions running for more than one month might help researches to draw significant results and reveal the long-term impact of the robots. Reference: Abdi J, Al-Hindawi A, Ng T, Vizcaychipi MPJBo. Scoping review on the use of socially assistive robot technology in elderly care. 2018;8(2):e018815.

Encapsulation and Outdoor Testing of Perovskite Solar Cells: Comparing Industrially Relevant Process with a Simplified Lab Procedure

Perovskite solar cells (PSCs) have shown great potential for next-generation photovoltaics. One of the main barriers to their commercial use is their poor long-term stability under ambient conditions and, in particular, their sensitivity to moisture and oxygen. Therefore, several encapsulation strategies are being developed in an attempt to improve the stability of PSCs in a humid environment. The lack of common testing procedures makes the comparison of encapsulation strategies challenging. In this paper, we optimized and investigated two common encapsulation strategies: lamination-based glass-glass encapsulation for outdoor operation and commercial use (COM) and a simple glue-based encapsulation mostly utilized for laboratory research purposes (LAB). We compare both approaches and evaluate their effectiveness to impede humidity ingress under three different testing conditions: on-shelf storage at 21 °C and 30% relative humidity (RH) (ISOS-D1), damp heat exposure at 85 °C and 85% RH (ISOS-D3), and outdoor operational stability continuously monitoring device performance for 10 months under maximum power point tracking on a roof-top test site in Berlin, Germany (ISOS-O3). LAB encapsulation of perovskite devices consists of glue and a cover glass and can be performed at ambient temperature, in an inert environment without the need for complex equipment. This glue-based encapsulation procedure allowed PSCs to retain more than 93% of their conversion efficiency after 1566 h of storage in ambient atmosphere and, therefore, is sufficient and suitable as an interim encapsulation for cell transport or short-term experiments outside an inert atmosphere. However, this simple encapsulation does not pass the IEC 61215 damp heat test and hence results in a high probability of fast degradation of the cells under outdoor conditions. The COM encapsulation procedure requires the use of a vacuum laminator and the cells to be able to withstand a short period of air exposure and at least 20 min at elevated temperatures (in our case, 150 °C). This encapsulation method enabled the cells to pass the IEC 61215 damp heat test and even to retain over 95% of their initial efficiency after 1566 h in a damp heat chamber. Above all, passing the damp heat test for COM-encapsulated devices translates to devices fully retaining their initial efficiency for the full duration of the outdoor test (>10 months). To the best of the authors’ knowledge, this is one of the longest outdoor stability demonstrations for PSCs published to date. We stress that both encapsulation approaches described in this work are useful for the scientific community as they fulfill different purposes: the COM for the realization of prototypes for long-term real-condition validation and, ultimately, commercialization of perovskite solar cells and the LAB procedure to enable testing and carrying out experiments on perovskite solar cells under noninert conditions.
Keywords: IEC damp heat test; encapsulation; outdoor testing; perovskite solar cell; stability.

An ultra-low background alpha detection system with a Micromegas-based time projection chamber

In this paper, a time projection chamber (TPC) system for α radiation detection with ultra-low background is presented, which is based on Micromegas, a type of micro-pattern gaseous detector. Based on multi-dimensional information acquired by the TPC, such as the features of particle energy deposition and 3D track, the system can achieve the purpose of α identification with ultra-low background. A prototype TPC with the volume of 15 × 15 × 6.5 cm3 was developed. It was filled with a gas mixture of 95% neon and 5% isobutene at about 1 bar, and no additional shielding or control of alpha-emitting materials was employed. A low-noise high-integration electronic system based on multi-channel waveform sampling techniques was adopted to read out the anode strip signals. A test with a 241Am α source (5.5 MeV), a 49-h background test, and several other tests with weak radioactive samples in the laboratory environment were conducted. A feature selection method was designed to maximize the suppression of background events while retaining the events of interest. Test results showed that within an assumed 5 × 5 cm2 effective area in the center region of the TPC window, the background count rate of this system was lower than 1.6 × 10-3 counts per minute at 95% confidence level, while retaining the acceptance rate of about 96% for 241Am α particles.

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