Miracle-dz/UniFAR-FTUs huggingface dataset:
Exploration of CRISPR/Cas9-based gene editing as therapy for osteoarthritis

Objectives Osteoarthritis (OA) is a painful and debilitating disease and it is associated with aberrant upregulation of multiple factors, including matrix metalloproteinase 13 (MMP13), interleukin-1β (IL-1β) and nerve growth factor (NGF). In this study, we aimed to use the CRISPR/Cas9 technology, a highly efficient gene-editing tool, to study whether the ablation of OA-associated genes has OA-modifying effects. Methods We performed intra-articular injection of adeno-associated virus, which expressed CRISPR/Cas9 components to target each of the genes encoding MMP13, IL-1β and NGF, in a surgically induced OA mouse model. We also tested triple ablations of NGF, MMP13 and IL-1β. Results Loss-of-function of NGF palliates pain but worsens joint damage in the surgically induced OA model. Ablation of MMP13 or IL-1β reduces the expression of cartilage-degrading enzymes and attenuates structural deterioration. Targeting both MMP13 and IL-1β significantly mitigates the adverse effects of NGF blockade on the joints. Conclusions CRISPR-mediated ablation of NGF alleviates OA pain, and deletion of MMP13-1β or IL-1β attenuates structural damage in a post-traumatic OA model. Multiplex ablations of NGF, MMP13 and IL-1β provide benefits on both pain management and joint structure maintenance. Our results suggest that CRISPR-based gene editing is useful for the identification of promising drug targets and the development of feasible therapeutic strategies for OA treatment.
Preclinical Evaluation and Pilot Clinical Study of Al18F-PSMA-BCH for Prostate Cancer PET Imaging

Prostate-specific membrane antigen (PSMA)–targeted radioligands have played an important role in the diagnosis of prostate cancer. In this study, we developed an Al18F-labeled radiotracer and evaluated its potential for prostate cancer imaging. Methods: Al18F-PSMA-BCH (BCH is Beijing Cancer Hospital) was efficiently prepared manually. The binding affinity to PSMA was evaluated in vitro using the 22Rv1 (PSMA-positive) cell line. Small-animal PET imaging, biodistribution studies of Al18F-PSMA-BCH in mice bearing 22Rv1 and PC-3 (PSMA-negative) xenografted tumors, and a comparison with 68Ga-PSMA-617 in mice bearing LNCaP tumors were performed. PET/CT imaging was performed on 11 newly diagnosed prostate cancer patients at 1 and 2 h after injection. Biodistribution and preliminary efficacy were evaluated, and radiation dosimetry was estimated using OLINDA/EXM 2.0 software. Results: Al18F-PSMA-BCH was prepared within 30 min and was found to bind to PSMA with a dissociation constant of 2.90 ± 0.83 nM. Small-animal PET imaging of Al18F-PSMA-BCH could clearly differentiate 22Rv1 tumors from PC-3 tumors, as confirmed by ex vivo biodistribution data (7.87% ± 2.37% and 0.54% ± 0.22% injected dose/g at 1 h in 22Rv1 and PC-3 tumors, respectively). The uptake of Al18F-PSMA-BCH in 22Rv1 tumors could be substantially blocked by excess ZJ-43, a PSMA inhibitor. High-level accumulation of Al18F-PSMA-BCH was observed in PSMA-expressing organs, with increased uptake at later time points. Thirty-seven tumor lesions were detected in 11 patients, and the SUVmax in 27 lesions increased between 1 and 2 h after injection (10.60 vs. 14.11). The SUVmax in primary lesions in patients with high-risk prostate cancer was higher than that in patients with intermediate-risk prostate cancer. The kidneys received the highest estimated dose, 0.135 mGy/MBq, and the effective dose was 0.016 mGy/MBq. Conclusion: Al18F-PSMA-BCH was conveniently prepared with a reasonable yield within 30 min and showed a promising imaging capability for prostate cancer with reasonable radiation exposure. Al18F-PSMA-BCH can be used for prostate cancer imaging as a novel 18F PET radiotracer.
Improving Reliability of Myocontrol Using Formal Verification

In the context of assistive robotics, myocontrol is one of the so-far unsolved problems of upper-limb prosthetics. It consists of swiftly, naturally, and reliably converting biosignals, non-invasively gathered from an upper-limb disabled subject, into control commands for an appropriate self-powered prosthetic device. Despite decades of research, traditional surface electromyography cannot yet detect the subject’s intent to an acceptable degree of reliability, that is, enforce an action exactly when the subject wants it to be enforced. In this paper, we tackle one such kind of mismatch between the subject’s intent and the response by the myocontrol system, and show that formal verification can indeed be used to mitigate it. Eighteen intact subjects were engaged in two target achievement control tests in which a standard myocontrol system was compared to two “repaired” ones, one based on a non-formal technique, thus enforcing no guarantee of safety, and the other using the satisfiability modulo theories (SMT) technology to rigorously enforce the desired property. The experimental results indicate that both repaired systems exhibit better reliability than the non-repaired one. The SMT-based system causes only a modest increase in the required computational resources with respect to the non-formal technique; as opposed to this, the non-formal technique can be easily implemented in existing myocontrol systems, potentially increasing their reliability.
Impact of modified gelatin on valvular microtissues

A significant challenge in the field of tissue engineering is the biofabrication of three‐dimensional (3D) functional tissues with direct applications in organ‐on‐a‐chip systems and future organ engineering. Multicellular valvular microtissues can be used as building blocks for the formation of larger scale valvular macrotissues. Yet, for the controlled biofabrication of 3D macrotissues with predefined complex shapes, directed assembly of microtissues through bioprinting is needed. This study aimed to investigate if modified gelatin is an instructive material for valvular microtissues. Valvular microtissues were encapsulated in modified gelatin hydrogels and cross‐linked in the presence of a photoinitiator (Irgacure 2959 or VA‐086). Hydrogel properties were determined, and valvular interstitial cell functions like phenotype, proliferation, migration, mRNA expression of extracellular matrix (ECM) molecules, ECM deposition, and tissue fusion were characterized by histochemical stainings and RT‐qPCR. Encapsulated microtissues remained viable, produced heart valve‐related ECM components, and remained in a quiescent state. However, encapsulation induced some changes in ECM formation and gene expression. Encapsulated microtissues showed lower remodeling capacity and increased expression levels of Col I/V, elastin, hyaluronan, biglycan, decorin, and Sox9 compared with nonencapsulated microtissues. Furthermore, this study demonstrated that proliferation, migration, and tissue fusion was more pronounced in softer gels. In general, we evidenced that modified gelatin is an instructive material for physiologically relevant valvular microtissues and provided a proof of concept for the formation of larger valvular tissue by assembling microtissues at random in soft gels.
Caught Between Expectations and the Practice Field.

BACKGROUND Volunteer crisis line responders are a valuable resource for suicide prevention crisis lines worldwide. AIM The aim of this study was to gain a deeper understanding of how volunteers operating a diaconal crisis line in Norway experienced challenges and how these challenges were met. METHOD A qualitative, explorative study was conducted. A total of 27 volunteers were interviewed through four focus groups. The material was analyzed using systematic text condensation. RESULTS The greatest challenge to the volunteers was the perception of a gap between their expectations and the practice field. The experience of many volunteers was that the crisis line primarily served a broad ongoing support function for loneliness or mental illness concerns, rather than a suicide prevention crisis intervention function. LIMITATIONS The focus group design may have made the participants more reluctant to share experiences representing alternative perspectives or personally sensitive information. CONCLUSION The findings of this study suggest that a uniform response to callers using crisis lines as a source of ongoing support is warranted and should be implemented in volunteer training programs.
Signal Adaptive System for Space/Spatial-Frequency Analysis

This paper outlines the development of a multiple-clock-cycle implementation (MCI) of a signal adaptive two-dimensional (2D) system for space/spatial-frequency (S/SF) signal analysis. The design is based on a method for improved S/SF representation of the analyzed 2D signals, also proposed here. The proposed MCI design optimizes critical design performances related to hardware complexity, making it a suitable system for real time implementation on an integrated chip. Additionally, the design allows the implemented system to take a variable number of clock cycles (CLKs) (the only necessary ones regarding desirable—2D Wigner distribution-presentation of autoterms) in different frequency-frequency points during the execution. This ability represents a major advantage of the proposed design which helps to optimize the time required for execution and produce an improved, cross-terms-free S/SF signal representation. The design has been verified by a field-programmable gate array (FPGA) circuit design, capable of performing S/SF analysis of 2D signals in real time.
Efficacy of systemic temozolomide‐activated phage‐targeted gene therapy in human glioblastoma

Glioblastoma multiforme (GBM) is the most lethal primary intracranial malignant neoplasm in adults and most resistant to treatment. Integration of gene therapy and chemotherapy, chemovirotherapy, has the potential to improve treatment. We have introduced an intravenous bacteriophage (phage) vector for dual targeting of therapeutic genes to glioblastoma. It is a hybrid AAV/phage, AAVP, designed to deliver a recombinant adeno‐associated virus genome (rAAV) by the capsid of M13 phage. In this vector, dual tumor targeting is first achieved by phage capsid display of the RGD4C ligand that binds the αvβ3 integrin receptor. Second, genes are expressed from a tumor‐activated and temozolomide (TMZ)‐induced promoter of the glucose‐regulated protein, Grp78. Here, we investigated systemic combination therapy using TMZ and targeted suicide gene therapy by the RGD4C/AAVP‐Grp78. Firstly, in vitro we showed that TMZ increases endogenous Grp78 gene expression and boosts transgene expression from the RGD4C/AAVP‐Grp78 in human GBM cells. Next, RGD4C/AAVP‐Grp78 targets intracranial tumors in mice following intravenous administration. Finally, combination of TMZ and RGD4C/AAVP‐Grp78 targeted gene therapy exerts a synergistic effect to suppress growth of orthotopic glioblastoma.
Novel Magnetic Nanostructured Beads for Cadmium(II) Removal

This study presents an effective magnetic separation method for cadmium removal, based on the use of a novel nanostructured material as an adsorbent. This adsorbent involves the incorporation of magnetite nanoparticles (Fe3O4-NPs), synthesized by the reverse coprecipitation method, into sodium alginate and activated carbon to form spherical structures by crosslinking Ca2+ ions with the charged alginate chains, referred to as magnetic alginate activated carbon (MAAC) beads. The effect of the experimental parameters, such as pH, contacting time, adsorbent dosage, agitation type, and rotating speed were investigated and optimized for an efficient removal of Cd(II) ions at an initial concentration of 250 mg/L. The amount of adsorbed Cd(II) by MAAC beads increased at a pH of 6 with a removal efficiency over 90%. The maximum adsorption capacity reached was 70 mg/g of adsorbent at an initial Cd(II) concentration of 150 mg/L, whereas at 250 mg/L the adsorption capacity lowered until 60 mg/g. Sorption isotherms were calculated using Langmuir, Freundlich, Temkin, and Dubinin–Radushkevich equations, and were better described by the Freundlich and Temkin models. These results proved the removal efficiency and the potential use under real environmental conditions of the MAAC beads, due to their easy recovery from contaminated aqueous solutions.
First Report of Phytoplasmas in Soybean, Alfalfa, and Lupinus sp. in Lithuania.

Plants of cultivated soybean (Glycine max) and alfalfa (Medicago sativa) in Dotnuva and of wild Lupinus sp. in Ledakalnis, Lithuania, exhibited symptoms that suggested phytoplasmal infections. Soybean plants were of normal growth habit but exhibited veinal necrosis. Alfalfa and Lupinus plants exhibited stunting, abnormally small leaves, and witches'-broom symptoms. Diseases in the plants were termed soybean veinal necrosis (SVN), alfalfa stunt (AlfS), and Lupinus stunt (LupS), respectively. The presence of phytoplasmas in diseased plants was assessed using polymerase chain reaction (PCR) for amplification of phytoplasma-specific 16S rDNA. A phytoplasma-characteristic 1.2-kbp DNA fragment was amplified from all diseased plants but not from known healthy plants in nested PCRs in which the first DNA amplification was primed by primer pair P1/P7 and reamplification of DNA was primed by primer pair F2n/R2 (2,4). Products from the nested PCR primed by F2n/R2 were subjected to restriction fragment length polymorphism (RFLP) analysis, and the RFLP patterns obtained were compared with patterns previously published (1-4). On the basis of AluI, HaeIII, HhaI, HpaI, KpnI, MseI, and RsaI RFLP patterns, the SVN and LupS phytoplasmas were classified in group 16SrIII (peach X-disease phytoplasma group), subgroup B (III-B, type strain clover yellow edge phytoplasma), and the AlfS phytoplasma was classified in group 16SrI (aster yellows phytoplasma group), subgroup B (I-B, type strain aster yellows phytoplasma). Nucleotide sequences were determined for 16S rDNA fragments amplified from SVN and AlfS phytoplasmas in nested PCRs primed by F2n/R2. The sequences were deposited in GenBank under Accession nos. AF177383 for SVN and AF177384 for AlfS. Sequence similarity between the 16S rDNAs of SVN and Canadian clover yellow edge (strain CYE-C, GenBank Accession no. AF175304) phytoplasmas was 99.8%; sequence similarity between 16S rDNAs of AlfS and aster yellows (strain SAY, GenBank Accession no. M86340) phytoplasmas was 99.6%. The SVN phytoplasma 16S rDNA shared 100% sequence similarity with a 16S rDNA from the Lithuanian clover yellow edge (CYE-L, GenBank Accession no. AF173558) phytoplasma. The nucleotide sequence data supported the conclusion that the SVN and AlfS phytoplasmas were closely related to strains classified in subgroups III-B and I-B, respectively. Our findings extend the known geographic ranges of phytoplasma subgroups I-B and III-B to northern Europe, including Lithuania, and expand the known plant host ranges of these pathogens. References: (1) R. E. Davis et al. Int. J. Syst. Bacteriol. 47:262, 1997. (2) R. Jomantiene et al. Int. J. Syst. Bacteriol. 48:269, 1998. (3) R. Jomantiene et al. HortScience 33:1069, 1998. (4) I. -M. Lee et al. Int. J. Syst. Bacteriol. 48:1153, 1998.
Interleukin-8 Gene −251 A/T (rs4073) Polymorphism and Coronary Artery Disease Risk: A Meta-Analysis

Background Inflammation plays an important role in the pathogenesis of coronary artery disease (CAD). Studies have reported that inflammatory cytokine interleukin-8 (IL-8) gene −251 A/T (rs4073) polymorphism is correlated with CAD susceptibility, but the result remains controversial. The objective of this study was to clarify the association between IL-8 gene −251 A/T polymorphism and CAD risk. Material/Methods A meta-analysis included 8244 patients from 9 individual studies with 10 populations was conducted. Heterogeneity test was conducted, and pooled odds ratio (OR) with 95% confidence interval (CI) was calculated used fixed-effect or random-effects model accordingly. Publication bias was evaluated with the Begg’s funnel plot and Egger’s test. Sensitivity analysis was also conducted. Results A significant association between IL-8 gene −251 A/T polymorphism and CAD risk was found in the dominant model (OR 1.42, 95% CI 1.16–1.76, P<0.001), recessive model (OR 1.30, 95% CI 1.12–1.52, P<0.001), allelic model (OR 1.28, 95% CI 1.12–1.47, P<0.001), homozygote model (OR 1.59, 95% CI 1.21–2.08, P<0.001), and heterozygote model (OR 1.35, 95% CI 1.11–1.64, P=0.002). Subgroup analysis by ethnicity found significant associations in the Chinese population in the dominant model(OR 1.43, 95% CI 1.26–1.61, P<0.001), recessive model (OR 1.39, 95% CI 1.21–1.59, P<0.001), allelic model (OR 1.31, 95% CI 1.21–1.42, P<0.001), homozygote model (OR 1.66, 95% CI 1.41–1.95, P<0.001), and heterozygote model (OR 1.34, 95% CI 1.18–1.52, P<0.001), but no significant association was found in the Caucasian population. No significant publication bias was found. Conclusions The IL-8 gene −251 A/T polymorphism was significantly associated with CAD risk in the Chinese population but not in the Caucasian population, −251 A allele carrier had an increased risk of CAD in the Chinese population.
Electrospun polycaprolactone matrices with tensile properties suitable for soft tissue engineering

The extracellular environment is a complex network of functional and structural components that impart chemical and mechanical stimuli that affect cellular function and fate. Cell differentiation on three dimensional scaffolds is also determined by the modulus of the substrate. Electrospun PCL nanofibers, which mimic the extra cellular matrix, have been developed with a wide variety of solvents and their combinations. The various studies have revealed that the solvents used influence the physical and mechanical properties, resulting in scaffolds with Young's modulus in the range of 1.8–15.4 MPa, more suitable for engineering of hard tissue like bone. The current study describes the use of benign binary solvent-generated fibrous scaffolds with a Young's modulus of 36.05 ± 13.08 kPa, which is almost 50 times lower than that of scaffolds derived from the commonly used solvents, characterized with myoblast, which can be further explored for applications in muscle and soft tissue engineering.
On the networking synthesis of studio factors to the integration of design pedagogy

Studio is critically important for design education, but few attempts have been made to demonstrate the parallels between studio factors and design performance. This paper adopts a coherent set of analyses to investigate the major studio factors and attempts to quantify the networking interactions among them. First, it describes how architectural studio is usually organised based on some major factors. Next, a theoretical model is established according to the described hypotheses and their mutual interactions. Third, the research method and statistical analysis with structural equation modelling (SEM) are presented. Finally, the results of this empirical examination are presented for discussion and suggestions. Our findings reveal that studio tutorials have no significant effect on undergraduate's design performance. In contrast, students’ subjective intention plays a more important role in shaping their behaviour, indicating the importance of transferring those exterior forces into internal benefits when the studio instructor attempts to optimise the pedagogy. These findings are also inspiring for all creative disciplines.
Cervical and vaginal fluid soluble Toll-like receptor 2 in pregnancies complicated by preterm prelabor rupture of membranes

Abstract Objective: To determine the cervical and vaginal fluid soluble Toll-like receptor-2 (sTLR2) levels in pregnancies complicated by preterm prelabor rupture of membranes (PPROM) and their correlation to microbial invasion of the amniotic cavity (MIAC) and/or histological chorioamnionitis (HCA). Methods: Sixty-eight women with singleton pregnancies complicated by PPROM were included in this study. Cervical and vaginal fluid was collected at the time of admission, and levels of sTLR2 in the cervical and vaginal fluid were determined using enzyme-linked immunosorbent assay. Result: Women with MIAC and both MIAC and HCA did not have different cervical and vaginal fluid sTLR2 levels compared to those without MIAC and without both MIAC and HCA. Women with HCA had higher cervical fluid sTLR2 levels in crude analysis (with HCA: median 11.6 pg/mL versus without HCA: median 5.5 pg/mL; p = 0.04) but not after adjustment for gestational age at sampling (p = 0.19). No difference in vaginal fluid sTLR2 levels between women with and without HCA was found. A positive correlation between cervical and vaginal fluid sTLR2 levels was identified (rho = 0.54; p < 0.0001). Conclusions: Cervical and vaginal fluid sTLR2 levels did not reflect the presence of MIAC and/or HCA.
Complete mitochondrial genome of the Trichogaster leeri (Anabantoidei: Osphronemidae)

Abstract The complete mitochondrial genome of Trichogaster leeri is determined in this study. It is 16,454 bp in length and consists of 13 protein-coding genes, 2 rRNA genes, 22 tRNA genes and a non-coding control region (D-loop). The overall base composition of the heavy-strand (H-strand) of the T. trichopterus mitochondrial genome is A: 29.18%, T: 28.07%, C: 27.20% and G: 15.55%. The total length of the 13 protein-coding genes was 11,435 bp. Phylogenetic analysis showed that the species of Trichogaster (T. trichopterus, T. lalius and T. leeri) formed a monophyletic group and represented close relationship with the species of Anabantoidei. This study provides an important data set for phylogenetic and taxonomic analyses of the species of genus Trichogaster.
Monte Carlo Simulations of Water Exchange Through Myelin Wraps: Implications for Diffusion MRI

Diffusion magnetic resonance imaging (dMRI) yields parameters sensitive to brain tissue microstructure. A structurally important aspect of this microstructure is the myelin wrapping around the axons. This paper investigated the forward problem concerning whether water exchange via the spiraling structure of the myelin can meaningfully contribute to the signal in dMRI. Monte Carlo simulations were performed in a system with intra-axonal, myelin, and extra-axonal compartments. Diffusion in the myelin was simulated as a spiral wrapping the axon, with a custom number of wraps. Exchange (or intra-axonal residence) times were analyzed for various number of wraps and axon diameters. Pulsed gradient sequences were employed to simulate the dMRI signal, which was analyzed using different methods. Diffusional kurtosis imaging analysis yielded the radial diffusivity (RD) and radial kurtosis (RK), while the two-compartment Kärger model yielded estimates the intra-axonal volume fraction (<inline-formula> <tex-math notation="LaTeX">$\nu _{\mathrm{ ic}}$ </tex-math></inline-formula>) and exchange time (<inline-formula> <tex-math notation="LaTeX">$\tau $ </tex-math></inline-formula>). Results showed that <inline-formula> <tex-math notation="LaTeX">$\tau $ </tex-math></inline-formula> was on the sub-second level for geometries with axon diameters below <inline-formula> <tex-math notation="LaTeX">$1.0~\mu {\mathrm{ m}}$ </tex-math></inline-formula> and less than eight wraps. Otherwise, exchange was negligible compared to typical experimental durations, with <inline-formula> <tex-math notation="LaTeX">$\tau $ </tex-math></inline-formula> of seconds or longer. In situations where exchange influenced the signal, estimates of RK and <inline-formula> <tex-math notation="LaTeX">$ \nu _{\mathrm{ ic}}$ </tex-math></inline-formula> increased with the number of wraps, while RD decreased. <inline-formula> <tex-math notation="LaTeX">$\tau $ </tex-math></inline-formula> estimates from simulated signals were in agreement with predicted ones. In conclusion, exchange through spiraling myelin permits sub-second <inline-formula> <tex-math notation="LaTeX">$ \tau $ </tex-math></inline-formula> for small diameters and low number of wraps. Such conditions may arise in the developing brain or in neurodegenerative disease, and thus the results could aid the interpretation of dMRI studies.