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This review explores the potential benefits of fecal microbiota transplantation (FMT) as an adjunct treatment in tuberculosis (TB), drawing parallels from its efficacy in inflammatory bowel disease (IBD). FMT has shown promise in restoring the gut microbial balance and modulating immune responses in IBD patients. Considering the similarities in immunomodulation and dysbiosis between IBD and TB, this review hypothesizes that FMT may offer therapeutic benefits as an adjunct therapy in TB. Methods: We conducted a systematic review of the existing literature on FMT in IBD and TB, highlighting the mechanisms and potential implications of FMT in the therapeutic management of both conditions. The findings contribute to understanding FMT’s potential role in TB treatment and underscore the necessity for future research in this direction to fully leverage its clinical applications. Conclusion: The integration of FMT into the comprehensive management of TB could potentially enhance treatment outcomes, reduce drug resistance, and mitigate the side effects of conventional therapies. Future research endeavors should focus on well-designed clinical trials to develop guidelines concerning the safety and short- and long-term benefits of FMT in TB patients, as well as to assess potential risks.

The article investigates wound healing and fibrosis in intestinal disease. Although efforts for the development of anti-inflammatory treatments for patients with inflammatory bowel disease (IBD) have been extensive, treatments for fibrosis and fistulas in IBD are still lacking. Experts recommend for further studies to deepen understanding of the mechanisms responsible for the development of fibrosis in the gastrointestinal mucosa.

Nonintrusive load monitoring (NILM) can determine operating schedule of electrical loads in a target system from measurements made at a centralized location, such as the electric utility service entry. NILM is an ideal platform for extracting useful information about any system that uses electromechanical devices. It has a low installation cost and high reliability because it uses a bare minimum of sensors. It is possible to use modem state and parameter estimation algorithms to verify remotely the "health" of electromechanical loads by using NILM to analyze measured waveforms associated with the operation of individual loads. NILM can also monitor the operation of the electrical distribution system itself, identifying situations where two or more otherwise healthy loads interfere with each other's operation through voltage waveform distortion or power quality problems. Strategies for nonintrusive monitoring have developed over the last 20 years. Advances in computing technology make a new wealth of computational tools useful in practical, field-based NILM systems. This article reviews techniques for high-performance nonintrusive load and diagnostic monitoring and illustrates key points with results from field tests. [ABSTRACT FROM PUBLISHER]

Aggregate formation of superoxide dismutase 1 (SOD1) inside motor neurons is known as a major factor in onset of amyotrophic lateral sclerosis. The thermodynamic stability of the SOD1 β-barrel has been shown to decrease in crowded environments such as inside a cell, but it remains unclear how the thermodynamics of crowding-induced protein destabilization relate to SOD1 aggregation. Here we have examined the effects of a protein crowder, lysozyme, on fibril aggregate formation of the SOD1 β-barrel. We found that aggregate formation of SOD1 is decelerated even in mildly crowded solutions. Intriguingly, transient diffusive interactions with lysozyme do not significantly affect the static structure of the SOD1 β-barrel but stabilize an alternative excited "invisible" state. The net effect of crowding is to favor species off the aggregation pathway, thereby explaining the decelerated aggregation in the crowded environment. Our observations suggest that the intracellular environment may have a similar negative (inhibitory) effect on fibril formation of other amyloidogenic proteins in living cells. Deciphering how crowded intracellular environments affect aggregation and fibril formation of such disease-associated proteins will probably become central in understanding the exact role of aggregation in the etiology of these enigmatic diseases.

The structural stability of proteins is found to markedly change upon their transfer to the crowded interior of live cells. For some proteins, the stability increases, while for others, it decreases, depending on both the sequence composition and the type of host cell. The mechanism seems to be linked to the strength and conformational bias of the diffusive in-cell interactions, where protein charge is found to play a decisive role. Because most proteins, nucleotides, and membranes carry a net-negative charge, the intracellular environment behaves like a polyanionic ( Z :1) system with electrostatic interactions different from those of standard 1:1 ion solutes. To determine how such polyanion conditions influence protein stability, we use negatively charged polyacetate ions to mimic the net-negatively charged cellular environment. The results show that, per Na + equivalent, polyacetate destabilizes the model protein SOD1 barrel significantly more than monoacetate or NaCl. At an equivalent of 100 mM Na + , the polyacetate destabilization of SOD1 barrel is similar to that observed in live cells. By the combined use of equilibrium thermal denaturation, folding kinetics, and high-resolution nuclear magnetic resonance, this destabilization is primarily assigned to preferential interaction between polyacetate and the globally unfolded protein. This interaction is relatively weak and involves mainly the outermost N-terminal region of unfolded SOD1 barrel . Our findings point thus to a generic influence of polyanions on protein stability, which adds to the sequence-specific contributions and needs to be considered in the evaluation of in vivo data.

In the cytosolic environment, protein crowding and Brownian motions result in numerous transient encounters. Each such encounter event increases the apparent size of the interacting molecules, leading to slower rotational tumbling. The extent of transient protein complexes formed in live cells can conveniently be quantified by an apparent viscosity, based on NMR-detected spin-relaxation measurements, that is, the longitudinal ( T 1 ) and transverse ( T 2 ) relaxation. From combined analysis of three different proteins and surface mutations thereof, we find that T 2 implies significantly higher apparent viscosity than T 1 . At first sight, the effect on T 1 and T 2 seems thus nonunifiable, consistent with previous reports on other proteins. We show here that the T 1 and T 2 deviation is actually not a inconsistency but an expected feature of a system with fast exchange between free monomers and transient complexes. In this case, the deviation is basically reconciled by a model with fast exchange between the free-tumbling reporter protein and a transient complex with a uniform 143 kDa partner. The analysis is then taken one step further by accounting for the fact that the cytosolic content is by no means uniform but comprises a wide range of molecular sizes. Integrating over the complete size distribution of the cytosolic interaction ensemble enables us to predict both T 1 and T 2 from a single binding model. The result yields a bound population for each protein variant and provides a quantification of the transient interactions. We finally extend the approach to obtain a correction term for the shape of a database-derived mass distribution of the interactome in the mammalian cytosol, in good accord with the existing data of the cellular composition.

Random encounters between proteins in crowded cells are by no means passive, but found to be under selective control. This control enables proteome solubility, helps to optimise the diffusive search for interaction partners, and allows for adaptation to environmental extremes. Interestingly, the residues that modulate the encounters act mesoscopically through protein surface hydrophobicity and net charge, meaning that their detailed signatures vary across organisms with different intracellular constraints. To examine such variations, we use in-cell NMR relaxation to compare the diffusive behaviour of bacterial and human proteins in both human and Escherichia coli  cytosols. We find that proteins that 'stick' in E. coli are generally less restricted in mammalian cells. Furthermore, the rotational diffusion in the mammalian cytosol is less sensitive to surface-charge mutations. This implies that, in terms of protein motions, the mammalian cytosol is more forgiving to surface alterations than E. coli cells. The cellular differences seem not linked to the proteome properties per se , but rather to a 6-fold difference in protein concentrations. Our results outline a scenario in which the tolerant cytosol of mammalian cells, found in long-lived multicellular organisms, provides an enlarged evolutionary playground, where random protein-surface mutations are less deleterious than in short-generational bacteria., Competing Interests: We declare no conflict of interest., (© 2020 The Author(s).)

In the disordered state, a protein exhibits a high degree of structural freedom, in both space and time. For an ensemble of disordered or unfolded proteins, this means that the ensemble comprises a high diversity of structures, ranging from compact collapsed states to fully extended polypeptide chains. In addition, each chain is highly dynamic and undergoes conformational changes and local dynamics on both fast and slow timescales. The size properties of disordered proteins are thus best described as ensemble averages. A straightforward measure of the size is the hydrodynamic radius, R H , of the ensemble. Since the disordered state is conformationally fluid, the observed R H does not refer to a particular shape or fold. Instead, it should be interpreted as a measure for the average compaction of the structural ensemble. In addition to characterizing the disordered ensemble itself, R H can be used to, with good precision, monitor changes in the ensemble size properties upon functional interactions of the disordered protein, e.g., dimerization, ligand binding, and folding pathways. Here, we present a step-by-step protocol for diffusion measurements using pulsed field gradient nuclear magnetic resonance (PFG NMR) spectroscopy. We describe how to calibrate the magnetic field gradient and offer different schemes for sample preparation. Finally, we describe how to obtain R H directly from the diffusion coefficient as well as from using an internal standard as a reference.

Aim: The aim of this study was to assess the prevalence of elevated transaminase levels in children undergoing prophylactic treatment for tuberculosis (TB) infection., Methods: All children living in a geographically defined area, who started TB prophylaxis during 2009-2011, were included. Data on background factors, treatment regimes and transaminase levels at baseline and follow-up were collected retrospectively., Results: Of the 277 children who were treated, 113 (41%) had elevated transaminase levels. Of these, 97 (35%) had levels that were less than three times the upper limit of the normal range and 16 (6%) had levels that were more than three times the normal range. Four patients had to discontinue isoniazid treatment and were successfully switched to rifampicin. In 17 patients, the highest transaminase peak did not occur until after 6 months of treatment. Elevated transaminases were significantly more common in patients below 9 years of age (62%) than in those aged 10-18 years (28%). Transaminases were elevated in 44% of all boys and 36% of all girls (p = 0.17)., Conclusion: Transaminase elevation was common in children receiving prophylactic treatment for TB and started at different points throughout the treatment period. Younger patients faced an increased risk. Regular blood tests are recommended throughout treatment., (©2015 Foundation Acta Paediatrica. Published by John Wiley & Sons Ltd.)

Expression in Escherichia coli represents the simplest and most cost effective means for the production of recombinant proteins. This is a routine task in structural biology and biochemistry where milligrams of the target protein are required in high purity and monodispersity. To achieve these criteria, the user often needs to screen several constructs in different expression and purification conditions in parallel. We describe a pipeline, implemented in the Center for Optimized Structural Studies, that enables the systematic screening of expression and purification conditions for recombinant proteins and relies on a series of logical decisions. We first use bioinformatics tools to design a series of protein fragments, which we clone in parallel, and subsequently screen in small scale for optimal expression and purification conditions. Based on a scoring system that assesses soluble expression, we then select the top ranking targets for large-scale purification. In the establishment of our pipeline, emphasis was put on streamlining the processes such that it can be easily but not necessarily automatized. In a typical run of about 2 weeks, we are able to prepare and perform small-scale expression screens for 20-100 different constructs followed by large-scale purification of at least 4-6 proteins. The major advantage of our approach is its flexibility, which allows for easy adoption, either partially or entirely, by any average hypothesis driven laboratory in a manual or robot-assisted manner.

Objectives: To test the ability of patients with brain injury to use a patient locator and minder (PLAM) system to assist in their adherence to therapy schedules., Participants: Five patients with acquired brain injury who were inpatients on an acute rehabilitation floor of a rehabilitation hospital., Measures: The number of human prompts necessary to direct a patient to, and ensure arrival at, a scheduled therapy destination and the proportion of therapy sessions requiring no prompting was measured both before and after the introduction of the PLAM system., Results: With the PLAM system, the average number of human prompts dropped by more than 50%, and the number of sessions requiring no prompting increased from 7% to 44%., Conclusion: The PLAM system described in this article seems feasible and useful for patients with acquired brain injury in assisting them with arrival at their therapy destinations without the assistance of staff.

Macrophages in lymphoid organs are in close contact to nerve terminals of the sympathetic nervous system. Hence, these cells could be targets of neuronal modulation. We studied sympathetic neurotransmitters as chemoattractants enabling the aggregation of macrophages and nerve terminals. Norepinephrine (NE), neuropeptide Y (NPY), isoproterenol (beta-adrenergic), p-aminoclonidine (alpha2-adrenergic), methoxamine (alpha1-adrenergic), and adenosine triphosphate (ATP) were used to study human monocyte and macrophage migration in 48-well Boyden chambers. NE stimulated chemotaxis of monocytes and macrophages at an optimal concentration of 10-(10) M (P < 0.025). Isoproterenol, but not p-aminoclonidine or methoxamine, induced chemotaxis of monocytes (10(-10) M, P < 0.05). In these studies, elevation of cAMP is a critical step in NE-induced chemotaxis of monocytes. NPY (10(-11) M, P < 0.05) stimulated monocyte chemotaxis as well. ATP at 10(-4) and 10(-5) M stimulated undirected cell mobility (P < 0.05). All tested neurotransmitters of the sympathetic nerve terminal were potent chemoattractants. These findings may explain the close association of nerves and macrophages in tissue and lymphoid organs and may thus be of functional relevance in neuroimmunomodulation.