Temporal Proteomic Profiling During Differentiation of Normal and Dystrophin-Deficient Human Muscle Cells

Background: Myogenesis is a dynamic process involving temporal changes in the expression of many genes. Lack of dystrophin protein such as in Duchenne muscular dystrophy might alter the natural course of gene expression dynamics during myogenesis. Objective: To gain insight into the dynamic temporal changes in protein expression during differentiation of normal and dystrophin deficient myoblasts to myotubes. Method: A super SILAC spike-in strategy in combination and LC-MS/MS was used for temporal proteome profiling of normal and dystrophin deficient myoblasts during differentiation. The acquired data was analyzed using Proteome Discoverer 2.2. and data clustering using R to define significant temporal changes in protein expression. Results: sFour major temporal protein clusters that showed sequential dynamic expression profiles during myogenesis of normal myoblasts were identified. Clusters 1 and 2, consisting mainly of proteins involved mRNA splicing and processing expression, were elevated at days 0 and 0.5 of differentiation then gradually decreased by day 7 of differentiation, then remained lower thereafter. Cluster 3 consisted of proteins involved contractile muscle and actomyosin organization. They increased in their expression reaching maximum at day 7 of differentiation then stabilized thereafter. Cluster 4 consisting of proteins involved in skeletal muscle development glucogenesis and extracellular remodeling had a lower expression during myoblast stage then gradually increased in their expression to reach a maximum at days 11–15 of differentiation. Lack of dystrophin expression in DMD muscle myoblast caused major alteration in temporal expression of proteins involved in cell adhesion, cytoskeleton, and organelle organization as well as the ubiquitination machinery. Conclusion: Time series proteome profiling using super SILAC strategy is a powerful method to assess temporal changes in protein expression during myogenesis and to define the downstream consequences of lack of dystrophin on these temporal protein expressions. Key alterations were identified in dystrophin deficient myoblast differentiation compared to normal myoblasts. These alterations could be an attractive therapeutic target.

An in vitro model for the cultivation of polymicrobial biofilms under continuous-flow conditions

The airways of people with cystic fibrosis (CF) are often chronically colonised with a diverse array of bacterial and fungal species. However, little is known about the relative partitioning of species between the planktonic and biofilm modes of growth in the airways. Existing in vivo and in vitro models of CF airway infection are ill-suited for the long-term recapitulation of mixed microbial communities. Here we describe a simple, in vitro continuous-flow model for the cultivation of polymicrobial biofilms and planktonic cultures on different substrata. Our data provide evidence for inter-species antagonism and synergism in biofilm ecology. We further show that the type of substratum on which the biofilms grow has a profound influence on their species composition. This happens without any major alteration in the composition of the surrounding steady-state planktonic community. Our experimentally-tractable model enables the systematic study of planktonic and biofilm communities under conditions that are nutritionally reminiscent of the CF airway microenvironment, something not possible using any existing in vivo models of CF airway infection.

Collection alteration and pristine structural properties of dust particles collected by MIDAS/Rosetta at comet 67P/Churyumov-Gerasimenko

<p>Comets are believed to have preserved pristine material from the early stages of the Solar System formation, thus providing unique information on intricate processes like dust growth mechanisms. The Rosetta mission gave us the best opportunity to investigate nearly pristine cometary dust particles of comet 67P/Churyumov–Gerasimenko. Among the three in-situ dust instruments, the MIDAS (Micro-Imaging Dust Analysis System) atomic force microscope collected cometary dust particles with sizes from hundreds of nanometres to tens of micrometres and recorded their 3D topography, size, shape, morphology, and related parameters [1].</p> <p>MIDAS collected dust emitted from comet 67P on dedicated targets. Particles fell through the entry funnel and collided with the collection targets [2] causing an unknown degree of particle alteration. To understand which structural properties of the dust remained pristine and can be used to understand comets and early Solar System processes it is important to understand the collection alteration. Dedicated laboratory experiments were carried out by previous studies [3, 4]. They found that the degree of alteration upon collection is strongly determined by the particle size, strength, and the collection velocity. They indicate that particles in the MIDAS size range deposited with moderate velocities about less than a few metres per second can stick on a target without major alteration.</p> <p>We aim to determine the structurally least altered MIDAS particles and investigate their properties. As database we use an improved version of the MIDAS particle catalogue [5]. Selecting all particles suitable for our analysis (e.g., cometary origin, sufficiently high image quality) grants us topographic data of over 600 nano- to micrometre-sized dust particles of comet 67P. We create dust coverage maps showing the distribution of the selected dust particles on the collection targets. As first, simple classification we divide the particles into those detected in clusters, suggested to be fragments originating in a shattering event of one large parent particle, and those remote from others that are potentially individually collected particles. Finally, we use a shape descriptor to categorise the particles according to their characteristics, e.g., shape and size, and compare to previous results from COSIMA [6] and simulation/laboratory studies [3, 7].</p> <p> </p> <p>[1] Bentley, M.S., Schmied, R., Mannel, T., et al. 2016, Nature, 537</p> <p>[2] Bentley, M. S., Arends, H., Butler, B., et al. 2016, Acta Astronautica, 125, 11</p> <p>[3] Ellerbroek, L. E., Gundlach, B., Landeck, A., et al. 2017, MNRAS, 469, S204</p> <p>[4] Ellerbroek, L. E., Gundlach, B., Landeck, A., et al. 2019, MNRAS, 486, S3755</p> <p>[5] Boakes, P., and the MIDAS team, 2018. ‘MIDAS Particle Catalogue’. ESA Planetary Science Archive  Dataset: RO-C-MIDAS-5-PRL-TO-EXT3-V2.0. Product ID: RO-C-MIDAS-5-PRL-TO-EXT3-V2.0</p> <p>[6] Langevin, Y., Hilchenbach, M., Ligier, N., et al. 2016, Icarus, 271, 76</p> <p>[7] Lasue, J., Maroger, I., Botet, R., et al. 2019, A&A, 630,</p>

A Low Energy Approach for the Preparation of Nano-Emulsions with a High Citral-Content Essential Oil

Pectis elongata is found in the northern and northeastern regions of Brazil. It is considered a lemongrass due to its citric scent. The remarkable citral content and the wide antimicrobial properties and bioactive features of this terpene make this essential oil (EO) eligible for several industrial purposes, especially in cosmetics and phytotherapics. However, to address the problems regarding citral solubility, nano-emulsification is considered a promising strategy thanks to its improved dispersability. Thus, in this paper we propose a low-energy approach for the development of citral-based nano-emulsions prepared with P. elongata EO. The plant was hydrodistillated to produce the EO, which was characterized with a gas chromatograph coupled to mass spectrometry. The nano-emulsion prepared by a non-heated water titrating (low-energy) method was composed of 5% (w/w) EO, 5% (w/w) non-ionic surfactants and 90% (w/w) deionized water and was analyzed by dynamic light scattering. Levels of citral of around 90% (neral:geranial—4:5) were detected in the EO and no major alteration in the ratio of citral was observed after the nano-emulsification. The nano-emulsion was stable until the 14th day (size around 115 nm and polydispersity index around 0.2) and no major alteration in droplet size was observed within 30 days of storage. Understanding the droplet size distribution as a function of time and correlating it to concepts of compositional ripening, as opposing forces to the conventional Ostwald ripening destabilization mechanism, may open interesting approaches for further industrial application of novel, low-energy, ecofriendly approaches to high citral essential oil-based nano-emulsions based on lemongrass plants.

LOSS OF MASTICATORY APPARATUS INTEGRITY - DISABILITY OR DISABLEMENT?

Были обследованы 408 пациентов (151 мужчина и 257 женщин) 54-85 лет (средний возраст 73,9±4,9 года) с помощью клинических (опрос, осмотр, пальпация, перкуссия, аускультация), социологических («GOHAI»), параклинических (определение эффективности жевания, чистоты речи, исследование дефицита массы тела, ортопантомография, КТ) методов. Все пожилые пациенты были с двумя патологическими состояниями - декомпенсированным зубным рядом и полной потерей зубов. У 322 человек рабочий синдромологический диагноз - декомпенсированный зубной ряд - относился к 167 нижним челюстям, 65 верхним челюстям и у 90 человек - и к верхним, и к нижним челюстям (всего 412 зубных рядов). Эффективность жевания у контингента беззубых также была невысокой, несмотря на наличие у большинства из них съемных зубных протезов. Согласно гериатрическому показателю здоровья полости рта («GOHAI»), ответы обследуемых колебались от 7 до 25 баллов (в среднем 18,23±4,19 балла), что относилось к низкому уровню. Общими синдромами пациентов оказались грубые, уродующие, старящие изменения внешнего вида лица, значительное нарушение эстетических норм. Не менее важны инвалидизирующие факторы почти полного отсутствия способности к жеванию, звукообразования, речи, затруднённого проглатывания пищи. В силу явных морфофункциональных и психосоциальных нарушений, социальной дезадаптации, сопровождающих эти заболевания, таких пациентов следовало бы считать инвалидами для повышения доступности геронтостоматологической реабилитации и включения их в программу государственных гарантий оказания стоматологической ортопедической помощи. Необходимо также ввести квотирование имплантационного зубного протезирования для пациентов пожилого и старческого возраста, которое по своей результативности и эффективности вписывается в систему адекватной реабилитации указанного контингента лиц и является мерой профилактики инвалидности. We examined 408 patients (151 male, 257 female) in an age range from 54 to 85 years (mean age 73,9±4,9) performing clinical (anamnesis, visual analysis, palpation, percussion, auscultation) sociological (GOHAI), additional (chewing efficiency evaluation, phonetic analysis, weight deficit of body calculation, OPG, CBCT) assessment. Among 322 of examined subjects working syndromological diagnosis was «decompensated dentition» related to lower jaw in 167 cases, 65 to upper jaw, 90 to both upper and lower jaws (altogether 412 dental arches). Chewing efficiency among edentulous patients was low considering that majority of patients had dentures. Score range for Geriartric General Oral Health Assessment Index (GOHAI) was from 7 to 25 points (mean 18,23±4,19) which is attributed as low level. General syndromes among patients were rough, disfiguring, ageing changes in facial appearance, major alteration of aesthetic norms. Equally important or even more was incapacitation impact of total loss of proper mastication, speech problems, phonetic alteration, problematic food swallowing. In our research we highlighted two pathological conditions among elderly and senile age groups: decompensated dentition, and fully edentulous jaws. Taking into consideration suffering of the patients due to morphological functional and psychosocial disabling conditions, accompanying these diseases, we might consider this group of patients as handicapped and that could lead to the need to introduction of setting quotas of implant prosthetics treatment of the elderly and senile patients. The result, efficacy and efficiency of such rehabilitation are considered in compliance with the system of adequate treatment result for this part of population.

Degradation of the veterinary antibiotics in swine manure via anaerobic digestion

Antimicrobial resistance is becoming an imminent danger for human health, in part because of excessive consumption of antibiotics in the animal industry. Laboratory-scale anaerobic digesters (ADs) were utilized to investigate the potential removal of veterinary antibiotics (VA) in swine manure. Nine laboratory ADs were kept at 39 [plus-minus] 2 [degrees] C in incubators and were loaded every two days. The digesters had a working volume of 1.38 L (in a 1.89-L glass jar), with a hydraulic retention time (HRT) of 21 days and a loading rate of 1.0 g-VS/L-day. Carbon dioxide concentration of the biogas and pH values of the digestate verifying digester performance were measured every four days and every week, respectively. The AD process allowed 2 HRT to stabilize before the addition of the VAs, of Chlortetracycline (CTC), Tylosin, and Sulfamethazine (SMZ). Tests were conducted to compare the effects of VAs on manure nutrients, volatile solid destruction, and biogas production. Concentrations of VA added to the manure samples were 263 to 298 mg/L of CTC, 88 to 263 mg/L of Tylosin, and 1042 to 1339 mg/L of SMZ, respectively. Analysis of VA concentrations before and after the AD process was conducted to determine the VA degradation. The process of measuring the relatively low VA concentrations involves using solid-phase extraction and LC-MS-MS methodology. Further tests were performed in water under room temperature and digester temperature to approve the degradation of VAs. Amplicon sequencing analysis performed on each group using the 50 most frequent amplicon sequence variants (ASVs) showed significant discriminations between groups. A hierarchical method (UPGMA) was applied to obtain the major alteration in bacterial dynamics between treatments. The biomarkers' abundance can be employed to distinguish different antibiotics contamination.

The Impact of Biodiesel Blend Variation Contamination to Engine Friction, Wear, Performance and Emission

Biodiesel can be utilised in diesel engines without major alteration to the engine; however, its use can lead to engine oil dilution through biodiesel leaking and scraping to the engine oil pan. The objective of this research is to investigate the contamination of biodiesel in engine lubricant oil, and determine the relationship between engine performance and emission for three different blends of palm methyl ester (B10, B20, and B30). To simulate the contamination, the engine oils were blended with biodiesel fuels at 5%, 10%, 15% and 20% by volume and these mixtures were tested on four-ball test equipment. An air-cooled diesel engine was used to analyse the influence of the three biodiesel blends on the output and emission of the diesel engine. The results show that both coefficient of friction and wear scar diameter increased with the increase of biodiesel percentage. The results of engine performance during both sweep test and step test showed that the variation of torque and power among the blends at a particular speed was within a very narrow range.

Pregnancy-induced changes in the transcriptome of the bovine corpus luteum during and after embryonic interferon-tau secretion

Understanding luteal maintenance during early pregnancy is of substantial biological and practical importance. Characterizing effects of early pregnancy, however, has historically been confounded by use of controls with potential exposure to early PGF pulses or differences in CL age. To avoid this, the present study utilized bihourly blood sampling to ensure control CL (n = 6) were of a similar age to CL from pregnant animals (n = 5), yet without exposure to PGF pulses. Additionally, CL from second month of pregnancy (n = 4) were analyzed to track fate of altered genes after cessation of embryonic interferon tau (IFNT) secretion. The major alteration in gene expression in first month of pregnancy occurred in interferon-stimulated genes (ISGs), with immune/interferon signaling pathways enriched in three independent over-representation analyses. Most ISGs decreased during second month of pregnancy, though, surprisingly, some ISGs remained elevated in the second month even after cessation of IFNT secretion. Investigation of luteolytic genes found few altered transcripts, in contrast to previous reports, likely due to removal of controls exposed to PGF pulses. An exception to this trend was decreased expression of transcription factor NR4A1. Beyond luteolytic genes and ISGs, over representation analyses highlighted the prevalence of altered genes within the extracellular matrix and regulation of IGF availability, confirming results of other studies independent of luteolytic genes. These results support the idea that CL maintenance in early pregnancy is related to lack of PGF exposure, although potential roles for CL expression of diverse ISGs and other pathways activated during early pregnancy remain undefined.

Impact of wetting/drying cycles on the hydromechanical behaviour of a treated soil

The positive effects of lime or cement treatment could be altered by weathering in the very long term. In this context, the main purpose of this study is to examine the impact of wetting/drying cycles on the strength and the hydraulic conductivity of a compacted soil treated with lime and cement. Compacted specimens were cured for 90 days before being exposed up to twelve wetting and drying cycles. A special concern of the study was the experimental method to impose the wetting and drying cycles. Two protocols were employed: one relied on relative humidity control to dry the samples, while the other was based on oven drying. The impact of the cycles was quantified by comparing the performance of the samples exposed to the cycles to the performance of the unsolicited samples. The results showed that the cycles induced a major alteration of the strength of the samples, with both methods. This degradation is associated to a significant increase of the hydraulic conductivity of the samples with the number of cycles.

Antimicrobial Study of Clinical Mastitic Milk Samples of Cows in and Around Bikaner

Mastitis, an inflammatory condition of the mammary gland, is one of the most economically important diseases of dairy herds. This disease is more prevalent in high yielding dairy cattle. It is caused due to colonization of pathogenic bacteria causing series of events that lead to major alteration in the mammary tissues and composition of milk secreted from the tissue cells (Batavani et al., 2007; Goel et al., 2008). Clinical mastitis is recognized by abnormal milk, varying degree of mammary gland inflammation (redness, heat, swelling and pain) with or without illness of the cow. Mastitis in dairy animals leads to heavy economic losses due to reduction in milk production, altered composition, and cost of treatment of affected animals. The clinical mastitis in different cases can be identified by external symptoms such as udder swelling, touch, pain, redness and hardness of the udder. The objective of the present study was to assess the sensitivity of bacterial isolates from mastitic milk samples of cows to some selected antibiotics.