Miracle-dz/UniFAR-FTUs huggingface dataset:
Site-Selective C–H Halogenation Using Flavin-Dependent Halogenases Identified via Family-Wide Activity Profiling

Enzymes are powerful catalysts for site-selective C–H bond functionalization. Identifying suitable enzymes for this task and for biocatalysis in general remains challenging, however, due to the fundamental difficulty of predicting catalytic activity from sequence information. In this study, family-wide activity profiling was used to obtain sequence–function information on flavin-dependent halogenases (FDHs). This broad survey provided a number of insights into FDH activity, including halide specificity and substrate preference, that were not apparent from the more focused studies reported to date. Regions of FDH sequence space that are most likely to contain enzymes suitable for halogenating small-molecule substrates were also identified. FDHs with novel substrate scope and complementary regioselectivity on large, three-dimensionally complex compounds were characterized and used for preparative-scale late-stage C–H functionalization. In many cases, these enzymes provide activities that required several rounds of directed evolution to accomplish in previous efforts, highlighting that this approach can achieve significant time savings for biocatalyst identification and provide advanced starting points for further evolution.
Computerised decision support to aid primary care clinicians in the management of specialist drugs : an evaluation of the needs of UK General Practitioners (GPs)

2 In the UK since the early 1990s there has been a growing trend for certain hospital initiated specialist drugs to be prescribed by GPs within primary care. GPs have been encouraged to undertake this activity supported by the use of so-called “shared care protocols. ” Despite these arrangements this has proven difficult with concerns over the quality and availability of shared care protocols and reported safety related incidents including fatalities linked to the use of certain specialist drugs. GPs have benefited from the complete computerisation of general practice unlike their hospital based colleagues who on the whole continue to work and prescribe within paper-based systems. These benefits have included the use of electronic prescribing and the introduction of computerised decision support systems (CDSS) in the form of reminders and drug related warnings and alerts. Scoping reviews are an established way of assessing the evidence base of a subject area which can then be summarised to reflect the broad nature of the field. For this thesis a scoping review of CDSS was used to evaluate how this intervention has been used within primary care and to identify areas where it can be further developed. Although the use of information technology has been suggested as a possible solution to some of the problems at the primary and secondary care interface, no published studies have evaluated the potential for a CDSS to support the prescribing of specialist drugs by GPs. In order to explore this concept further and gain additional knowledge of current CDSS usage, key informants were identified and interviewed. The key informants described the current availability and characteristics of CDSS within UK general practice. The concept of developing a CDSS to support GPs in the use of specialist drugs to include prescribing was acknowledged as beneficial and was widely supported. Enablers and barriers to development and implementation were identified including a number of potential operating models. Key enablers included data quality and functionality features, joint development and implementation and to make use of existing systems and frameworks. Key barriers included addressing the needs of end users, security, regulation and funding. Human ergonomics was used to further investigate GPs and the actual level and use of computers and software programs including CDSS at the point of prescribing both during and outside of patient consultations. The application of an analytical approach to these processes through a task analysis framework identified failings in existing arrangements for GPs to safely prescribe specialist drugs.
Mobile Geriatric Teams – A Cost-Effective Way Of Improving Patient Safety And Reducing Traditional Healthcare Utilization Among The Frail Elderly? A Randomized Controlled Trial

Background Demographic changes combined with costly technological progress put a financial strain on the healthcare sector in the industrialized world. Hence, there is a constant need to develop new cost-effective treatment procedures in order to optimize the use of available resources. As a response, the concept of a Mobile Geriatric Team (MGT) has emerged not only nationally but also internationally during the last decade; however, scientific evaluation of this initiative has been very scarce. Thus, the objective of this study was to perform a mixed methods analysis, including a prospective, controlled and randomized quantitative evaluation, in combination with an interview-based qualitative assessment, to measure the effectiveness and user satisfaction of MGT. Materials and methods Community-dwelling, frail elderly people were randomized to an intervention group (n=31, mean age 84) and a control group (n=31, mean age 86). A two-year retrospective quantitative data collection and a prospective one-year follow-up on healthcare utilization were combined with qualitative interviews. Non-parametric statistics and difference-in-difference (DiD) analyses were applied to the quantitative data. Qualitative data were analyzed using content analysis. Results No significant group differences in healthcare utilization were found before inclusion. Post intervention, primary care contact (including MGTs) increased for the MGT group. Inpatient care decreased dramatically for both groups. Hence, the increase in primary care contact for MGT patients was not accompanied by a reduction in inpatient care compared to the control group. Utilization of non-primary care was lower (p< 0.01) post-intervention in both groups. Conclusion There appears to be a “natural” variation in healthcare needs over time among frail elderly people. Hence, it is vital to perform open, controlled clinical studies in tandem with the implementation of new caregiving strategies. The MGT initiative was clearly appreciated but did not fully achieve the desired reduction in healthcare utilization in this study. Trial registration Retrospectively registered 09/10/2018, ClinicalTrials.gov ID NCT03662945.
Expression And Biological Interaction Network Of RHOC For Hepatic Carcinoma With Metastasis In PBMC Samples

Objectives Hepatic carcinoma with metastasis remains incurable, and clinical diagnostic methods lacked adequate sensitivity and specificity. Therefore, seeking effectively diagnostic biomarkers is still essential for it. RHOC was reported to be linked to metastasis of hepatic carcinoma. However, almost all of the studies used tissues as detection samples, which was not ideal for clinical course minoring. Therefore, here, it was aimed to use PBMC samples that were not only easily accessible but also minimally invasive to determine the expression and biological interaction network of RHOC for hepatic carcinoma with metastasis. Methods PBMC samples were isolated. Then, RNA-seq was performed to identify the DEGs between hepatic carcinoma with metastasis and hepatic carcinoma with solitary tumor. Subsequently, q-RT-PCR was used to verify the expression level of RHOC. Finally, bioinformatic analysis was used to present the biological interaction network of RHOC for hepatic carcinoma with metastasis in PBMC samples. Results The results of both RNA-seq and q-RT-PCR showed that the expression level of RHOC was significantly higher in the PBMC samples of hepatic carcinoma with metastasis than in those of hepatic carcinoma with solitary tumor. By using variety of bioinformatic analysis platforms, in PBMCs, 18 co-expression genes with RHOC were identified and their interaction network showed that MYL9 and RHOC had the highest edge evidence, and were involved in some cell migration-related pathways. Conclusion Our results indicated that RHOC in PBMCs could be potentially minimally invasive indicators for the diagnosis and clinical course supervision of hepatic carcinoma with metastasis, and its biological interaction network determined based on bioinformatic methods would lay a foundation for further study of the role of RHOC in tumor invasion and metastasis.
Distinct roles of GRIN2A and GRIN2B variants in neurological conditions

Rapid advances in sequencing technology have led to an explosive increase in the number of genetic variants identified in patients with neurological disease and have also enabled the assembly of a robust database of variants in healthy individuals. A surprising number of variants in the GRIN genes that encode N-methyl-D-aspartate (NMDA) glutamatergic receptor subunits have been found in patients with various neuropsychiatric disorders, including autism spectrum disorders, epilepsy, intellectual disability, attention-deficit/hyperactivity disorder, and schizophrenia. This review compares and contrasts the available information describing the clinical and functional consequences of genetic variations in GRIN2A and GRIN2B. Comparison of clinical phenotypes shows that GRIN2A variants are commonly associated with an epileptic phenotype but that GRIN2B variants are commonly found in patients with neurodevelopmental disorders. These observations emphasize the distinct roles that the gene products serve in circuit function and suggest that functional analysis of GRIN2A and GRIN2B variation may provide insight into the molecular mechanisms, which will allow more accurate subclassification of clinical phenotypes. Furthermore, characterization of the pharmacological properties of variant receptors could provide the first opportunity for translational therapeutic strategies for these GRIN-related neurological and psychiatric disorders.
R T ] 2 5 O ct 2 01 9 Categorified canonical bases and framed BPS states

We consider a cluster variety associated to a triangulated surface without punctures. The algebra of regular functions on this cluster variety possesses a canonical vector space basis parametrized by certain measured laminations on the surface. To each lamination, we associate a graded vector space, and we prove that the graded dimension of this vector space gives the expansion in cluster coordinates of the corresponding basis element. We discuss the relation to framed BPS states inN = 2 field theories of class S.
Therapeutic Management of Ketosis Associated with Sub-clinical Mastitis in Transition Cows

Present research records the association between sub-clinical mastitis and sub-clinical ketosis in apparently healthy early lactating cows. All cows were subjected to california mastitis test and cultural tests for confirmation of sub-clinical mastitis. Subclinical ketosis was identified based on serum BHBA levels (more than 1.40 mmol/L). Out of 62 cattle with subclinical mastitis, 28 were found to be having sub-clinical ketosis. In vitro drug sensitivity test was conducted on 62 whole milk cultures which revealed that sensitive to Enrofloxacin and Ceftriaxone with Tazobactum. 28 animals were divided into two groups and therapy was given with antibiotics (Enrofloxacin and Ceftriaxone with Tazobactam) along with Meloxicam and Chlorpheniramine maleate for both groups. For group-A cows were treated with additional supplementation of oral glucogenic precursor with Cyanocobalamin and Nicotinamide. After treatment both groups of animals were tested for serum BHBA levels and milk for cultural examination at 1st week and 2nd week of post therapy. Significant reduction in serum BHBA values recorded in group-A animals than compare with group-B animals after completion of therapy. Present study reports the effect of oral gluconeogenic precursor, cyanocobalamin and nicotinamide in the treatment of subclinical ketosis in cows.
Modulation of GABAA receptor function by PKA and PKC protein phosphorylation

Protein kinase A (PKA) has been reported to regulate synaptic αβγ GABAA receptor currents, but whether or not PKA regulates GABAA receptor periand extrasynaptic tonic currents is unknown. GABAA receptors containing α4 subunits are important in mediating tonic inhibition and exist as both α4βδ and α4βγ receptors in the brain. To mimic GABA-independent and GABA-dependent tonic currents, we transfected HEK 293T cells with α4β3δ or α4β3γ2L subunits and recorded spontaneous currents in the absence of applied GABA and steady-state currents in the presence of 1 μM GABA. Both α4β3δ and α4β3γ2L receptors displayed spontaneous currents, but PKA-activation increased spontaneous α4β3δ currents substantially more than spontaneous α4β3γ2L currents. The increase in spontaneous α4β3δ currents was due to an increase in single channel open frequency. In contrast, PKA-activation did not alter steady state, tonic currents recorded in the presence of 1 μM GABA. We concluded that PKA had a GABA concentration-dependent effect on α4β3δ and α4β3γ2L currents. In the absence of GABA, spontaneous α4β3δ, and to a lesser 35 extent α4β3γ2L currents, could provide a basal, tonic current that could be regulated by PKA. With increasing concentrations of extracellular GABA, however, tonic α4β3δ and α4β3γ2L currents would become more GABA-dependent and less PKAsensitive. Thus, in brain regions with fluctuating extracellular GABA levels, the dynamic range of GABA-activated tonic currents would be set by PKA, and the increase in tonic current produced by increasing GABA would be reduced by PKA mediated phosphorylation. When ambient GABA reaches micromolar concentrations, PKA would have no effect on steady-state tonic currents. Introduction GABAA receptors are the major inhibitory neurotransmitter receptors in the adult mammalian CNS and mediate both phasic and tonic inhibition (Farrant and Nusser 2005). Phasic inhibition is produced primarily by rapid, synchronous activation of post-synaptic αβγ receptors with brief pre-synaptic release of high concentrations of GABA, and in contrast, tonic inhibition is produced by persistent activation of periand extra-synaptic αβδ receptors at low concentrations of ambient GABA (Essrich et al. 1998; Nusser et al. 1998; Semyanov et al. 2004) as well as some αβγ receptors that are located outside of synapses (Caraiscos et al. 2004; Zhang et al. 2007). Tonic currents play critical roles in regulating neuronal excitability. Time-averaged net charge transferred by perior extrasynaptic GABAA receptors can be larger than time-averaged charge transferred by phasic currents conducted by synaptic receptors. In addition to GABA-activated tonic currents, there is also
Modelling low-cycle fatigue tests using a gradient-enhanced continuum damage model

This study is concerned with the modelling of uniaxial low-cycle fatigue tests using a gradient-enhanced continuum damage model. The aim of the paper is to show that a simple but adequate description can be performed using such kind of phenomenological approach. The model accounts for the coupling between plasticity and damage, including a mathematically consistent description of the softening behaviour and size dependency. It successfully predicts low-cycle fatigue and can deal with regular cycles. A consistent procedure to identify experimentally the constitutive parameters that appear in the theory is proposed. Lifetime predictions for different loading amplitudes are compared with experimental data for two aluminium alloys showing a good agreement. The one-dimensional model can be interpreted as a particular version of a more general gradient-enhanced damage model, which is summarized at the end of the paper.
Nitrogen Starvation of Assessment in the Production of Single Cell Oils and Biodiesel Quality in Heterotrophic Cultures of Cyanobacteria Phormidium autumnale

The cyanobacteria have great potential for aquaculture industries, bioproducts, bioenergy and bioremediation, as in the reduction of ammonia, phosphorus and organic compounds. The manipulation of the medium and the modes of operation of the cultures are necessary to induce the production of biomass and bioproducts, such as lipids. The aim of the study was to evaluate the different nitrogen sources and their depletion in heterotrophic cultures of the cyanobacterium Phormidium autumnale aimed at obtaining high yields lipid. The heterotrophic cultures were carried out in bubble column bioreactor employing cassava starch as organic carbon source. In the first stage, were studied different nitrogen source under the C/N (carbon/nitrogen) ratios in the culture medium of 20, 40 and 60. In the second stage, selected to nitrogen source with the greatest potential for the production of lipids and held its depletion searching for the induction of a cellular stress. The fatty acids profile in the end of cultures was used in the estimation of biodiesel quality. In the first stage, the cultures employing sodium nitrate under CN ratio of 60 resulting in lipid content of 13.22% and lipid productivity of 7.62 mg/L.h. From the depletion of sodium nitrate, second stage, obtained a lipid productivity and lipid content of 10.43 mg/L.h and 25.07%, respectively. The fatty acid profile showed, in the presence of nitrogen, saturated fatty acid fraction and monounsaturated of 76.72% and 23.88%, respectively. The depletion of sodium nitrate induced a change in the lipid profile, directing the concentration of saturated fatty acids of 98.97%. The biodiesel properties obtained meet the standards set by standardization of the quality of biodiesel.
Digital Health in Cardiology: The Estonian Perspective

Background: Digital health is rapidly entering clinical practice in cardiology. Estonia is one of the leading nations in implementing digital nationwide solutions. Recently, the European Society of Cardiology organized the 1st Summit on Digital Health in Tallinn, which provided the opportunity to discuss various aspects concerning the digitalization of cardiology. Summary: The current review focuses on the advancements of Estonian digital health and digital cardiology as well as possible barriers and solutions for implementing digital innovations in cardiology. Key Messages: The authors have included aspects from the recent summit, personal communications, and literature reviews to express the current state and future possibilities of digital health in cardiology from the Estonian perspective.
Long-Term Follow-Up and Oncological Outcome of Patients Undergoing Radical Cystectomy for Bladder Cancer following an Enhanced Recovery after Surgery (ERAS) Protocol: Results of a Large Randomized, Prospective, Single-Center Study

Introduction: Enhanced recovery after surgery (ERAS) concepts are implemented in various surgical disciplines. For patients undergoing radical cystectomy, prospective data are still rare. In the year 2014, our group could prove significant benefits of ERAS for these patients in a prospective randomized study compared to a conservative regimen. Objective: To evaluate long-term follow-up results of ERAS concepts in patients undergoing radical cystectomy for bladder cancer. Methods: Of the 101 patients who had initially prospectively been randomized to ERAS or a conservative regimen, 35 patients could be included in the study. Median follow-up time was 83 months. Primary end point assessed quality of life. Secondary end points were cancer-specific survival and overall survival. Results: Five to seven years after the initial inclusion to the study, no significant difference regarding quality of life parameters was detected (p values range between 0.112 and 0.970). Continence status in patients with neobladder showed no significant differences between ERAS and conservative regimens (p = 0.785). Cancer-specific survival in the ERAS group did not differ significantly from that in the conservative group (49 vs. 58%, p = 0.725). Conclusions: While ERAS represents an excellent way to improve postoperative reconvalescence and quality of life in the short-term follow-up, our data do not support the idea that there is also a long-term effect in terms of quality of life issues.
Timing of Morphine Administration Differentially Alters Paraventricular Thalamic Neuron Activity

Abstract The paraventricular thalamic nucleus (PVT) is a brain region involved in regulating arousal, goal-oriented behaviors, and drug seeking, all key factors playing a role in substance use disorder. Given this, we investigated the temporal effects of administering morphine, an opioid with strongly addictive properties, on PVT neuronal function in mice using acute brain slices. Here, we show that morphine administration and electrophysiological recordings that occur during periods of animal inactivity (light cycle) elicit increases in PVT neuronal function during a 24-h abstinence time point. Furthermore, we show that morphine-induced increases in PVT neuronal activity at 24-h abstinence are occluded when morphine administration and recordings are performed during an animals’ active state (dark cycle). Based on our electrophysiological results combined with previous findings demonstrating that PVT neuronal activity regulates drug-seeking behaviors, we investigated whether timing morphine administration with periods of vigilance (dark cycle) would decrease drug-seeking behaviors in an animal model of substance use disorder. We found that context-induced morphine-seeking behaviors were intact regardless of the time morphine was administered (e.g., light cycle or dark cycle). Our electrophysiological results suggest that timing morphine with various states of arousal may impact the firing of PVT neurons during abstinence. Although, this may not impact context-induced drug-seeking behaviors.
CRISPR interference (CRISPRi) as transcriptional repression tool for Hungateiclostridium thermocellum DSM 1313

Hungateiclostridium thermocellum DSM 1313 has biotechnological potential as a whole‐cell biocatalyst for ethanol production using lignocellulosic renewable sources. The full exploitation of H. thermocellum has been hampered due to the lack of simple and high‐throughput genome engineering tools. Recently in our research group, a thermophilic bacterial CRISPR–Cas9‐based system has been developed as a transcriptional suppression tool for regulation of gene expression. We applied ThermoCas9‐based CRISPR interference (CRISPRi) to repress the H. thermocellum central metabolic lactate dehydrogenase (ldh) and phosphotransacetylase (pta) genes. The effects of repression on target genes were studied based on transcriptional expression and product formation. Single‐guide RNA (sgRNA) under the control of native intergenic 16S/23S rRNA promoter from H. thermocellum directing the ThermodCas9 to the promoter region of both pta and ldh silencing transformants reduced expression up to 67% and 62% respectively. This resulted in 24% and 17% decrease in lactate and acetate production, correspondingly. Hence, the CRISPRi approach for H. thermocellum to downregulate metabolic genes can be used for remodelling of metabolic pathways without the requisite for genome engineering. These data established for the first time the feasibility of employing CRISPRi‐mediated gene repression of metabolic genes in H. thermocellum DSM 1313.