Miracle-dz/UniFAR-FTUs huggingface dataset:
Unroofing site-specific α-synuclein–lipid interactions at the plasma membrane

Significance α-Synuclein is a neuronal protein with an ill-defined biological function that is central to Parkinson’s disease etiology. While considered to be involved in exocytosis, how α-synuclein facilitates synaptic vesicle fusion and release remains an open question. To address this, we investigated α-synuclein–lipid interactions at the plasma membrane through the technique of cellular unroofing, which uncovers an intact basal membrane. We conclusively show that α-synuclein is recruited to exocytic sites, preferring liquid-ordered lipid domains. Importantly, heterogeneous populations of α-synuclein conformers are revealed by measurements of fluorescence lifetime distributions, which are not adequately described by current models of α-synuclein structures. Membrane-bound α-synuclein is conformationally dynamic, exquisitely sensitive to lipid/protein composition, enabling the protein to carry out its function. Parkinson’s disease is associated with α-synuclein (α-syn), a cytosolic protein enriched in presynaptic terminals. The biological function of α-syn remains elusive; however, increasing evidence suggests that the protein is involved in the regulation of synaptic vesicle fusion, signifying the importance of α-syn–lipid interactions. We show that α-syn preferentially binds to GM1-rich, liquid-ordered lipid domains on cytoplasmic membranes by using unroofed cells, which encapsulates lipid complexity and cellular topology. Moreover, proteins (Rab3a, syntaxin-1A, and VAMP2) involved in exocytosis also localize with α-syn, supporting its proposed functional role in exocytosis. To investigate how these lipid/protein interactions influence α-syn at the residue level, α-syn was derivatized with an environmentally sensitive fluorophore (7-nitrobenz-2-oxa-1,3-diazol-4-yl [NBD]) at different N- and C-terminal sites. Measurements of NBD fluorescence lifetime distributions reveal that α-syn adopts a multitude of membrane-bound conformations, which were not recapitulated in simple micelle or vesicle models, indicating an exquisite sensitivity of the protein to the complex lipid environment. Interestingly, these data also suggest the participation of the C terminus in membrane localization, which is generally overlooked and thus emphasize the need to use cellularly derived and biologically relevant membranes for biophysical characterization. Collectively, our results demonstrate that α-syn is more conformationally dynamic at the membrane interface than previously appreciated, which may be important for both its physiological and pathological functions.
Phylogenetic and Physiological Diversity of Cultured Deep-Biosphere Bacteria from Equatorial Pacific Ocean and Peru Margin Sediments

During ODP Leg 201 microbial communities in Eastern Equatorial Pacific Ocean and Peru Margin sediments were investigated. The sediment layers sampled extended down to 420 m below the sea floor, with estimated ages of up to 40 million years. Contamination-free anoxic slurries were inoculated into media containing different substrate combinations, all at micromolar concentration. These culture media were designed for a broad spectrum of physiological groups. A total of 162 pure cultures were isolated that could be grouped into 19 different phylotypes based on 16S rRNA gene analysis. The isolates belonged to the Alpha-, Gamma- and Deltaproteobacteria, the Firmicutes, Actinobacteria, and Bacteroidetes. The genera most frequently isolated were Bacillus (68 isolates) and Rhizobium (40 isolates). Comparison of strains with the same phylotypes by enterobacterial repetitive intergenic consensus (ERIC-PCR) analysis revealed the presence of several subgroups that did not correlate with medium, sediment depth or sampling site. The majority of the isolates, although obtained from anoxic environments and isolated under strictly anoxic conditions, turned out to be facultativly aerobic. Physiologically, the isolates were characterized as generalists, able to utilize a broad variety of electron donors with either oxygen, nitrate and in some cases manganese oxides as electron acceptors. The diversity inferred from physiological tests was even higher than that on the phylogenetic or genomic level. The outcome of the contamination tests, the isolation of close relatives of already known subsurface bacteria, the repeated finding of the same phylotype from different sites and the level of diversity present in the culture collection strongly suggest that indigenous deep-biosphere bacteria had been isolated.
Adult B‐ and T‐lymphoblastic lymphoma treated with a paediatric acute lymphoblastic leukaemia regimen have excellent outcomes—a short report from two Sydney centres

Armin Rashidi Thomas Kaiser Maryam Ebadi Shernan G. Holtan Tauseef Ur Rehman Daniel J. Weisdorf Alexander Khoruts Christopher Staley Division of Hematology, Oncology, and Transplantation, Department of Medicine, University of Minnesota, Minneapolis, Department of Surgery, University of Minnesota, Minneapolis, and Division of Gastroenterology, Hepatology, and Nutrition, Department of Medicine, University of Minnesota, Minneapolis, MN, USA. E-mail: arashidi@umn.edu
Applied kinesiology and dentistry – A narrative review

ABSTRACT Objective To investigate the use of applied kinesiology in the field of dentistry. Methods A review of the literature was carried out looking for all articles written on the topic. PubMed, Ovid Medline, and The Cochrane Central Register of Controlled Trials (CENTRAL) databases were searched. Results Only one study was retrieved on the use of applied kinesiology in temporomandibular disorder patients, and only one study was published on dental material testing. A change in muscle strength associated with changes in dental occlusion has been observed in many of the articles selected. Discussion The use of applied kinesiology for the diagnosis and treatment of pathologies in the field of dentistry is not supported by scientific evidence. However, a relationship between dental occlusion or maxillo-mandibular relationship and isometric muscle strength has been noted.
Low-Dose Hsp90 Inhibitor Selectively Radiosensitizes HNSCC and Pancreatic Xenografts

Purpose: Treatment approaches using Hsp90 inhibitors at their maximum tolerated doses (MTDs) have not produced selective tumor toxicity. Inhibition of Hsp90 activity causes degradation of client proteins including those involved in recognizing and repairing DNA lesions. We hypothesized that if DNA repair proteins were degraded by concentrations of an Hsp90 inhibitor below those required to cause nonspecific cytotoxicity, significant tumor-selective radiosensitization might be achieved. Experimental Design: Tandem mass tagged-mass spectrometry was performed to determine the effect of a subcytotoxic concentration of the Hsp90 inhibitor, AT13387 (onalespib), on global protein abundance. The effect of AT13387 on in vitro radiosensitization was assessed using a clonogenic assay. Pharmacokinetics profiling was performed in mice bearing xenografts. Finally, the effect of low-dose AT13387 on the radiosensitization of three tumor models was assessed. Results: A subcytotoxic concentration of AT13387 reduced levels of DNA repair proteins, without affecting the majority of Hsp90 clients. The pharmacokinetics study using one-third of the MTD showed 40-fold higher levels of AT13387 in tumors compared with plasma. This low dose enhanced Hsp70 expression in peripheral blood mononuclear cells (PBMCs), which is a biomarker of Hsp90 inhibition. Low dose monotherapy was ineffective, but when combined with radiotherapy, produced significant tumor growth inhibition. Conclusions: This study shows that a significant therapeutic ratio can be achieved by a low dose of Hsp90 inhibitor in combination with radiotherapy. Hsp90 inhibition, even at a low dose, can be monitored by measuring Hsp70 expression in PBMCs in human studies.
How Many Targeted Biopsy Cores are Needed for Clinically Significant Prostate Cancer Detection during Transperineal Magnetic Resonance Imaging Ultrasound Fusion Biopsy?

PURPOSE To uncover the optimal number of transperineal magnetic resonance imaging ultrasound (MRI/US) fusion targeted biopsy (TB) cores per lesion needed for detection of clinically significant prostate cancer (csPCa). MATERIALS AND METHODS A total of 101 patients with at least 1 lesion with a PI-RADS (Prostate Imaging Reporting and Data System) score of 3 or greater were recruited prospectively. At least 4 transperineal MRI/US fusion TB cores per lesion were performed, followed by systematic biopsy (SB). The Kappa test was used to evaluate the consistency of csPCa detection rate (CDR) between different TB cores and ≥4 cores which was regarded as reference standard. RESULTS In the total cohort of 101 patients, 49 (48.5%), 55 (54.5%), and 57 (56.4 %) were detected with csPCa by SB, TB, or TB+SB, respectively. As for the total of 161 lesions, the CDR based upon 1, 2, 3, or ≥4 TB cores was made in 27.3%, 32.9%, 37.3% and 39.1%, respectively. Three cores showed great consistency with ≥4 cores in CDR (Kappa coefficient of 0.961, P<0.001) with a sensitivity of 95.2% (95% confidence interval 85.8-98.8%), and only miss 3 lesions harboring csPCa. The similar results were obtained in cases with PI-RADS 3 or 4 or maximal diameter of < 1.5 cm. CONCLUSIONS This study demonstrated that three TBs per lesion were suitable during transperineal MRI/US fusion biopsy, especially for lesions of PI-RADS 3 or 4 or small lesions (maximal diameter < 1.5 cm), which may help to tailor targeted prostate biopsy procedures.
Interleaved intramuscular stimulation with minimally overlapping electrodes evokes smooth and fatigue resistant forces

Objective. It is known that multi-site interleaved stimulation generates less muscle fatigue compared to single-site synchronous stimulation. However, in the limited number of studies in which intramuscular electrodes were used, the fatigue reduction associated with interleaved stimulation could not consistently be achieved. We hypothesize that this could be due to the inability to place the intramuscular electrodes used in interleaved stimulation in locations that minimize overlap amongst the motor units activated by the electrodes. Our objective in the present study was to use independent intramuscular electrodes to compare fatigue induced by interleaved stimulation with that generated by synchronous stimulation at the same initial force and ripple. Approach. In the medial gastrocnemius muscle of an anesthetized rabbit (n = 3), ten intramuscular hook wire electrodes were inserted at different distances from the nerve entry. Overlap was measured using the refractory technique and only three electrodes were found to be highly independent. After ensuring that forces obtained by both stimulation modalities had the same ripple and magnitude, fatigue induced during interleaved stimulation across three independent distal electrodes was compared to that obtained by synchronously delivering pulses to a single proximal electrode. Main results. Contractions evoked by interleaved stimulation exhibited less fatigue than those evoked by synchronous stimulation. Twitch force recruitment curves collected from each of the ten intramuscular electrodes showed frequent intermediate plateaus and the force value at these plateaus decreased as the distance between the electrode and nerve entry increased. Significance. The results indicate that interleaved intramuscular stimulation is preferred over synchronous intramuscular stimulation when fatigue-resistant and smooth forces are desired. In addition, the results suggest that the large muscle compartments innervated by the primary intramuscular nerve branches give rise to progressively smaller independent compartments in subsequent nerve divisions.
Produtividade e qualidade de frutos de cultivares de maracujazeiro-amarelo com ou sem polinização artificial

The objective of this work was to evaluate the influence of artificial pollination on productivity and fruit quality of yellow passion fruit cultivars. Seven commercial cultivars of yellow passion fruit were evaluated in two experiments carried out in Tangara da Serra, MT, Brazil, in a randomized block design with five replicates and plots consisting of two rows of four plants. In the first experiment, pollination was natural, whereas in the second it was artificial, performed twice a week. Planting was done on January 11, 2010, and harvests occured from July 2010 to March 2011. Artificial pollination increase the productivity of all cultivars, but their response to the technique varied. When artificial pollination was used, the cultivars FB 100, FB 200 and BRS Ouro Vermelho had a higher productivity than IAC 275, IAC 277, BRS Sol do Cerrado e BRS Gigante Amarelo. Artificial pollination increases productivity, fruit mass, diameter and length of fruit, and pulp percentage, and it reduces peel thickness. Strong genotype by environment interaction may be occurring, as for the productivity of the evaluated cultivars.
In Vitro Susceptibility of Treponema pallidum subsp. pallidum to Doxycycline

Doxycycline is regarded as an effective therapy for early syphilis, and there is increasing interest in using doxycycline for prophylaxis of this infection. However, the MIC of doxycycline for Treponema pallidum subsp. pallidum has not been reported previously. In this study, an in vitro culture system was utilized to determine that the MIC of doxycycline is 0.06 to 0.10 μg/ml for four strains of T. pallidum subsp. ABSTRACT Doxycycline is regarded as an effective therapy for early syphilis, and there is increasing interest in using doxycycline for prophylaxis of this infection. However, the MIC of doxycycline for Treponema pallidum subsp. pallidum has not been reported previously. In this study, an in vitro culture system was utilized to determine that the MIC of doxycycline is 0.06 to 0.10 μg/ml for four strains of T. pallidum subsp. pallidum (Nichols, SS14, UW231B, and UW249B). The Nichols strain cultured in vitro with doxycycline was also tested for infectivity in rabbits, and the minimum bactericidal concentration (MBC) was found to be ≤0.1 μg/ml using this method. The low MIC and MBC values are consistent with the previously demonstrated clinical efficacy of doxycycline for the treatment of early syphilis. This study represents the first report of the in vitro susceptibility of T. pallidum to doxycycline, and the resulting information may be useful in the consideration of doxycycline for use in prevention of syphilis.
Phosphotransferase System Uptake and Metabolism of the β-Glucoside Salicin Impact Group A Streptococcal Bloodstream Survival and Soft Tissue Infection

Streptococcus pyogenes (group A Streptococcus [GAS]), a major human-specific pathogen, relies on efficient nutrient acquisition for successful infection within its host. The phosphotransferase system (PTS) couples the import of carbohydrates with their phosphorylation prior to metabolism and has been linked to GAS pathogenesis. In a screen of an insertional mutant library of all 14 annotated PTS permease (EIIC) genes in MGAS5005, the annotated β-glucoside PTS transporter (bglP) was found to be crucial for GAS growth and survival in human blood and was validated in another M1T1 GAS strain, 5448. ABSTRACT Streptococcus pyogenes (group A Streptococcus [GAS]), a major human-specific pathogen, relies on efficient nutrient acquisition for successful infection within its host. The phosphotransferase system (PTS) couples the import of carbohydrates with their phosphorylation prior to metabolism and has been linked to GAS pathogenesis. In a screen of an insertional mutant library of all 14 annotated PTS permease (EIIC) genes in MGAS5005, the annotated β-glucoside PTS transporter (bglP) was found to be crucial for GAS growth and survival in human blood and was validated in another M1T1 GAS strain, 5448. In 5448, bglP was shown to be in an operon with a putative phospho-β-glucosidase (bglB) downstream and a predicted antiterminator (licT) upstream. Using defined nonpolar mutants of the β-glucoside permease (bglP) and β-glucosidase enzyme (bglB) in 5448, we showed that bglB, not bglP, was important for growth in blood. Furthermore, transcription of the licT-blgPB operon was found to be repressed by glucose and induced by the β-glucoside salicin as the sole carbon source. Investigation of the individual bglP and bglB mutants determined that they influence in vitro growth in the β-glucoside salicin; however, only bglP was necessary for growth in other non-β-glucoside PTS sugars, such as fructose and mannose. Additionally, loss of BglP and BglB suggests that they are important for the regulation of virulence-related genes that control biofilm formation, streptolysin S (SLS)-mediated hemolysis, and localized ulcerative lesion progression during subcutaneous infections in mice. Thus, our results indicate that the β-glucoside PTS transports salicin and its metabolism can differentially influence GAS pathophysiology during soft tissue infection.
Educational Interventions to Improve Dental Anatomy Carving Ability of Dental Students: A Systematic Review

This systematic review aimed to identify the level of impact of educational strategies for teaching tooth carving on the carving ability of undergraduate dental students. The PubMed‐NCBI, Cochrane‐CENTRAL, LILACS, Ibecs, BBO, Web of Science, and Scopus databases were searched in May 2019, for randomized controlled trials (RCTs) and two‐arm non‐randomized studies of interventions (NRSI) addressing educational interventions toward the dental carving of undergraduate students. Studies from the year 2000 until the search date, written in English, Portuguese, and Spanish were included. Study screening and data extraction were performed in duplicate and blinded. The data were presented narratively, considering the dental carving ability of students the primary outcome. The risk of bias was assessed using the RoB tool 2.0 and ROBINS‐I, and the level of evidence was determined with GRADE. Of 3,574 studies, 6 were included, with 3 RCTs and 3 NRSIs. Very low level of evidence was provided from the NRSIs that flipped classroom (1 study; n = 140) and a student‐driven revised module (1 study; n = 264) improved the carving ability of students. Additionally, there was moderate evidence of online complementary material (1 RCT; n = 30) and reinforcement class improving the carving ability of students (1 RCT; n = 29). The replacement of traditional classes by an instructional DVD (1 RCT; n = 73) and assessment of carving projects through digital systems (1 NRSI; n = 79) did not enhance the carving ability of students. Study design, risk of bias, and imprecision downgraded the level of evidence. There was a very low to moderate evidence on the effectiveness of student‐driven educational approaches and complementary classes of dental anatomy in improving the dental carving ability of students.
Characterization of ancestral myosin XI from Marchantia polymorpha by heterologous expression in Arabidopsis thaliana

Summary Previous studies have revealed duplications and diversification of myosin XI genes between angiosperms and bryophytes; however, the functional differentiation and conservation of myosin XI between them remain unclear. Here, we identified a single myosin XI gene from the liverwort Marchantia polymorpha (Mp). The molecular properties of Mp myosin XI are similar to those of Arabidopsis myosin XIs responsible for cytoplasmic streaming, suggesting that the motor function of myosin XI is able to generate cytoplasmic streaming. In cultured Arabidopsis cells, transiently expressed green fluorescent protein (GFP)‐fused Mp myosin XI was observed as some intracellular structures moving along the F‐actin. These intracellular structures were co‐localized with motile endoplasmic reticulum (ER) strands, suggesting that Mp myosin XI binds to the ER and generates intracellular transport in Arabidopsis cells. The tail domain of Mp myosin XI was co‐localized with that of Arabidopsis myosin XI‐2 and XI‐K, suggesting that all these myosin XIs bind to common cargoes. Furthermore, expression of GFP‐fused Mp myosin XI rescued the defects of growth, cytoplasmic streaming and actin organization in Arabidopsis multiple myosin XI knockout mutants. The heterologous expression experiments demonstrated the cellular and physiological competence of Mp myosin XI in Arabidopsis. However, the average velocity of organelle transport in Marchantia rhizoids was 0.04 ± 0.01 μm s−1, which is approximately one‐hundredth of that in Arabidopsis cells. Taken together, our results suggest that the molecular properties of myosin XI are conserved, but myosin XI‐driven intracellular transport in vivo would be differentiated from bryophytes to angiosperms.
Impact of vitamin D receptor gene polymorphisms on blood lead levels in Thai lead exposed workers

Background: Although lead poisoning has aroused public concern, its toxicokinetics and mechanism of toxicity are still unclear. Vitamin D regulates calcium homeostasis via vitamin D receptor (VDR) and its polymorphisms cause variability of body calcium and bone density. VDR polymorphisms may correlate with the variability of body burden of lead. Objective: Determine if vitamin D receptor polymorphisms is associated with blood lead in Thai workers exposed to lead. Subjects and methods: Four hundred fifteen lead exposure workers were recruited in the study. Blood lead level was determined as a biomarker of lead. Five VDR polymorphisms (FokI, BsmI, Apa, TagI, and Cdx2) were studied. Results: The allele frequencies of F/f, B/b, A/a, T/t, and A/G for FokI, BsamI, ApaI, TagI and Cdx-2 polymorphisms were 059/0.41, 0.07/0.93, 0.32/0.68, 0.94/0.06 and 0.4/0.6, respectively. Gender, smoking status, lead exposure status, BsmI, and TaqI polymorphisms were associated with blood lead level. The BB genotype of BsmI and the tt genotype of TaqI have significantly lower blood lead levels than other genotypes of their polymorphisms. Conclusion: The BsmI and TaqI polymorphisms of the vitamin D receptor gene had impact on the blood lead level. This association was similar to their effects on calcium. Lead might share the same toxicokinetics with calcium.