Policy Learning for Actively Labeled Sample Selection on Lumbar Semi-supervised Classification.

Large labeled data bring significant performance improvement, but acquiring labeled medical data is particularly challenging due to the laborious, time-consuming, and medically qualified annotation. Semi-supervised learning has been employed to leverage unlabeled data. However, the quality and quantity of annotated data have a great influence on the performance of the semi-supervised model. Selecting informative samples through active learning is crucial and could improve model performance. Therefore, we propose a unified semi-supervised active learning architecture (RL-based SSAL) that alternately trains a semi-supervised network and performs active sample selection. Semi-supervised model is first well trained for sample selection, and selected label-required samples are annotated and added to the previously labeled dataset for subsequent semi-supervised model training. To learn to select the most informative samples, we adopt a policy learning-based approach that treats sample selection as a decision-making process. A novel reward function based on the product of predictive confidence and uncertainty is designed, aiming to select samples with high confidence and uncertainty. Comparisons with a semi-supervised baseline on collected lumbar disc herniation dataset demonstrate the effectiveness of the proposed RL-based SSAL, achieving over 3% promotion across different amounts of labeled data. Comparisons with other active learning methods and ablation studies reveal the superiority of proposed policy learning based on active sample selection and reward function. Our model trained with only 200 labeled data achieves an accuracy of 89.32% which is comparable to the performance achieved with the entire labeled dataset, demonstrating its significant advantage.

Long-term impact of nusinersen on motor and electrophysiological outcomes in adolescent and adult spinal muscular atrophy: insights from a multicenter retrospective study.

BACKGROUND: 5q spinal muscular atrophy (SMA) is a progressive autosomal recessive motor neuron disease. OBJECTIVE: We aimed to assess the effects of nusinersen on motor function and electrophysiological parameters in adolescent and adult patients with 5q SMA. METHODS: Patients with genetically confirmed 5q SMA were eligible for inclusion, and clinical data were collected at baseline (V1), 63 days (V4), 180 days (V5), and 300 days (V6). The efficacy of nusinersen was monitored by encompassing clinical assessments, including the Revised Upper Limb Module (RULM), Hammersmith Functional Motor Scale Expanded (HFMSE), 6-Minute Walk Test (6MWT), and percent-predicted Forced Vital Capacity in sitting position (FVC%) and Compound Muscle Action Potential (CMAP) amplitude. The patients were divided into "sitter" and "walker" subgroups according to motor function status. RESULTS: 54 patients were screened, divided into "sitter" (N = 22) and "walker" (N = 32), with the mean age at baseline of 27.03 years (range 13-53 years). The HFMSE in the walker subgroup increased significantly from baseline to V4 (mean change +2.32-point, P = 0.004), V5 (+3.09, P = 0.004) and V6 (+4.21, P = 0.005). The patients in both the sitter and walker subgroup had no significant changes in mean RULM between V1 and the following time points. Significant increases in CMAP amplitudes were observed in both upper and lower limbs after treatment. Also, patients with RULM ≥ 36 points showed significant CMAP improvements. Our analysis predicted that patients with CMAP amplitudes of trapezius ≥ 1.76 mV were more likely to achieve significant motor function improvements. CONCLUSIONS: Nusinersen effectively improves motor function and electrophysiological data in adolescent and adult patients with SMA. This is the first report on the CMAP amplitude changes in the trapezius after treatment in patients with SMA. The CMAP values effectively compensate for the ceiling effect observed in the RULM, suggesting that CMAP could serve as an additional biomarker for evaluating treatment efficacy.

Streak tube imaging lidar with kilohertz laser pulses and few-photons detection capability.

Lidar using active light illumination is capable of capturing depth and reflectivity information of target scenes. Among various technologies, streak tube imaging lidar (STIL) has garnered significant attention due to its high resolution and excellent precision. The echo signals of a STIL system using single laser pulse are often overwhelmed by noise in complex environments, making it difficult to discern the range of the target. By combining high-frequency laser pulses with the repetitive sweep circuit, the STIL system enables efficient detection of few-photons signal in weak-light environments. Additionally, we have developed a robust algorithm for estimating the depth and reflectivity images of targets. The results demonstrate that this lidar system achieves a depth resolution better than 0.5 mm and a ranging accuracy of 95 um. Furthermore, the imaging of natural scenes also validates the exceptional 3D imaging capability of this system.

Momordica charantia Is a Novel Host of 'Candidatus Phytoplasma malaysianum'-Related Strains Associated with Bitter Melon Stem Fasciation Disease in China.

Momordica charantia, also known as bitter melon, bitter gourd, and bitter squash, is a member of the Cucurbitaceae family and is widely grown in tropical and subtropical regions for its edible fruit and medicinal properties (Alves et al. 2017). In April 2022, bitter melon plants exhibiting stem fasciation and excessive tendril symptoms were observed in a 50-acre vegetable farm in Yijia Village, Weishan Yizu Huizu Autonomous County, Dali, Yunnan Province, China (Fig. 1). The farm primarily grew tomatoes, but around 400 bitter melon plants were planted in spots where tomatoes failed to establish. One plot had a 40% incidence rate, with four out of ten bitter melon plants showing symptoms. Scattered cases were observed in other plots, leading to an overall disease incidence rate of around 2% for the entire farm. Phytoplasma infection was suspected due to symptomatic plants growing in the same province as previously reported cases of phytoplasma diseases, such as happy tree (Camptotheca accuminata) witches'-broom disease, and the presence of phytoplasma-transmitting leafhoppers (Qiao et al. 2023). DNA was extracted from four symptomatic samples and two healthy controls collected from the abovementioned plot with a 40% disease incidence using Bioteke's Plant Genomic DNA Extraction Kit and then tested for phytoplasma infection. A nested PCR assay was conducted using primer pair P1/16S-SR followed by P1A/16S-SR to amplify the near full-length phytoplasma 16S rDNA (about 1.5kb) as previously described (Lee et al. 2004). None of the healthy controls tested positive for phytoplasma infection, while three out of four symptomatic plants showed positive results. The amplicons from the nested PCR were cloned into the pCRII-TOPO vector as previously described (Lee et al. 2004). The resulting clones were sequenced, and the representative sequence was deposited into GenBank (accession number PP489216). The iPhyClassifier (Zhao et al. 2009) was employed to determine the phytoplasma species and group/subgroup associated with the bitter melon stem fasciation (BMSF) disease. The results indicated that the diseased bitter melon plants were infected with a strain related to 'Candidatus Phytoplasma malaysianum' (EU371934), with a 98.07% sequence identity. The similarity coefficient was 1.00 compared to the reference strain of 16SrXXXII-D (GenBank accession: MW138004). The phytoplasma strain associated with BMSF disease was designated as BMSF1. In addition, the same DNA samples underwent further characterization of the BMSF strains. A nested PCR was conducted using primer pair rpL2F3/rpIR1A, followed by rp(III)-FN/rpIR1A to amplify a phytoplasma-specific rp gene segment (about 1.2 kb) (Martini et al. 2007; Davis et al. 2013). Three out of four samples tested positive, consistent with the 16S rRNA gene amplification results. Similarly, a primer pair L15F1/MapR1 followed by secYF1(III)/secYR1(III) was used to amplify a phytoplasma-specific partial spc operon (about 1.7 kb) that includes the complete secY gene and partial rpl15 and map genes, as previously described (Lee et al. 2010). The obtained rp and partial spc amplicons were cloned and sequenced (GenBank accession numbers PP464295 and PP464296). The rp and secY gene sequences were searched against the non-redundant nucleotide collection in the NCBI database using BLASTN. The top hit for the rp gene was 'Ca. Phytoplasma luffae' (CP054393), with 83.24% identity (1068/1283 base-matching). The top hit for the secY gene was also 'Ca. Phytoplasma luffae' (CP054393), with 72.53% identity (1294/1784 base-matching). The percent identity of the BMSF sequences compared to the top hit is low since no other group 16SrXXXII rp and secY gene sequences are available for comparison. A subgroup 16SrXXXII-D phytoplasma strain has been previously reported associated with Camptotheca acuminata witches'-broom (Qiao et al. 2023) and Trema tomentosa witches'-broom (Yu et al. 2021) in China. To our knowledge, bitter melon represents a new host of 'Candidatus Phytoplasma malaysianum'-related strains, and this is the first report of BMSF disease in China. The findings suggest that 'Candidatus Phytoplasma malaysianum'-related strains infect not only ornamental plants but also crops.

Semantic contrast with uncertainty-aware pseudo label for lumbar semi-supervised classification.

BACKGROUND: Lumbar disc herniation (LDH) is a prevalent spinal disease that can result in severe pain, with Magnetic resonance imaging (MRI) serving as a commonly diagnostic tool. However, annotating numerous MRI images, necessary for deep learning based LDH diagnosis, can be challenging and labor-intensive. Semi-supervised learning, mainly utilizing pseudo labeling and consistency regularization, can leverage limited labeled images and abundant unlabeled images. However, consistency regularization solely focuses on maintaining the semantic consistency of transformed unlabeled data but fails to utilize the semantic information from labeled data to guide the unlabeled data, and additionally, pseudo labeling is prone to confirmation bias. METHOD: We propose SeCoFixMatch, an innovative approach that seamlessly integrates semantic contrast and uncertainty-aware pseudo labeling into semi-supervised learning. Semantic contrast constraints the semantic consistency between labeled and unlabeled images. Pseudo labels are generated by combining predictive confidence and uncertainty, with uncertainty computing by optimizing the Kullback-Leibler (KL) loss between predictive and target Dirichlet distribution. RESULTS: Comparison with other semi-supervised models and ablation experiment with varying labeled data demonstrate the effectiveness and generalization of proposed model. Notably, SeCoFixMatch, trained with just 40 labels, outperforms the baseline model trained with 200 labels, reducing the annotation effort by a remarkable 80%. CONCLUSIONS: Proposed pseudo labeling algorithm generates more precise pseudo labels for semantic contrastive learning and semantic contrastive learning facilitates better feature representation, thereby further improving the prediction accuracy of pseudo label. The mutual reinforcement of pseudo labeling and semantic contrast constraints boosts the performance of semi-supervised algorithm.

Passive localization and reconstruction of multiple non-line-of-sight objects in a scene with a large visible transmissive window.

Passive non-line-of-sight imaging methods have been demonstrated to be capable of reconstructing images of hidden objects. However, current passive non-line-of-sight imaging methods have performance limitations due to the requirements of an occluder and aliasing between multiple objects. In this paper, we propose a method for passive localization and reconstruction of multiple non-line-of-sight objects in a scene with a large visible transmissive window. The analysis of the transport matrix revealed that more redundant information is acquired in a scene with a window than that with an occluder, which makes the image reconstruction more difficult. We utilized the projection operator and residual theory to separate the reconstruction equation of multiple objects into the independent equations of the located objects that can be reconstructed independently by TVAL3 and Split-Bregman algorithms, which greatly reduces the computational complexity of the reconstruction. Our method lays the foundation for multiple objects reconstruction in complex non-line-of-sight scenes.

Minisci-Type Dehydrogenative Coupling of N-Heteroaromatic Rings with Inert C(sp3)-H Enabled by a Visible-Light-Catalyzed Intermolecular Hydrogen Atom Transfer Process.

The Minisci-type dehydrogenative coupling of N-heteroaromatic rings with inert C-H or Si-H partners via visible-light-catalyzed hydrogen atom transfer has been reported. This methodology allows the coupling reactions to be carried out in water as a solvent under air atmospheric conditions with visible-light illumination. A wide range of inert C-H and Si-H partners could be directly coupled with various N-aromatic heterocycles to deliver products in good to excellent yields.

Diagnosis of Challenging Spinal Muscular Atrophy Cases with Long-Read Sequencing.

Spinal muscular atrophy (SMA) is an autosomal recessive neuromuscular disorder primarily caused by the deletion or mutation of the survival motor neuron 1 (SMN1) gene. This study assesses the diagnostic potential of long-read sequencing (LRS) in three patients with SMA. For Patient 1, who has a heterozygous SMN1 deletion, LRS unveiled a missense mutation in SMN1 exon 5. In Patient 2, an Alu/Alu-mediated rearrangement covering the SMN1 promoter and exon 1 was identified through a blend of multiplex ligation-dependent probe amplification, LRS, and PCR across the breakpoint. The third patient, born to a consanguineous family, bore four copies of hybrid SMN genes. LRS determined the genomic structures, indicating two distinct hybrids of SMN2 exon 7 and SMN1 exon 8. However, a discrepancy was found between the SMN1/SMN2 ratio interpretations by LRS (0:2) and multiplex ligation-dependent probe amplification (0:4), which suggested a limitation of LRS in SMA diagnosis. In conclusion, this newly adapted long PCR-based third-generation sequencing introduces an additional avenue for SMA diagnosis.

Screening and identification of functional bacterial attachment genes in aerobic granular sludge.

The screening and identification of attachment genes is important to exploring the formation mechanism of biofilms at the gene level. It is helpful to the development of key culture technologies for aerobic granular sludge (AGS). In this study, genome-wide sequencing and gene editing were employed for the first time to investigate the effects and functions of attachment genes in AGS. With the help of whole-genome analysis, ten attachment genes were screened from thirteen genes, and the efficiency of gene screening was greatly improved. Then, two attachment genes were selected as examples to further confirm the gene functions by constructing gene-knockout recombinant mutants of Stenotrophomonas maltophilia; when the two attachment genes were knocked out, the attachment potential was reduced by 50.67% and 43.93%, respectively. The results provide a new theoretical principle and efficient method for the development of AGS from the perspective of attachment genes.

Insights into refractory chronic inflammatory demyelinating polyneuropathy: a comprehensive real-world study.

Background: Refractory chronic inflammatory demyelinating polyneuropathy (CIDP) is a challenging subset of CIDP. It does not respond well to immune therapy and causes substantial disability. A comprehensive understanding of its clinical profile, electrophysiological characteristics and potential risk factors associated with refractoriness remains to be further elucidated. Methods: Data in this cross-sectional study was collected and reviewed from the Huashan Peripheral Neuropathy Database (HSPN). Included patients were categorized into refractory CIDP and non-refractory CIDP groups based on treatment response. The clinical and electrophysiological characteristics were compared between refractory and non-refractory CIDP groups. Potential risk factors associated with refractory CIDP were explored with a multivariate logistic regression model. Results: Fifty-eight patients with CIDP were included. Four disease course patterns of refractory CIDP are described: a relapsing-remitting form, a stable form, a secondary progressive form and a primary progressive form. Compared to non-refractory CIDP patients, refractory CIDP exhibited a longer disease duration (48.96 ± 33.72 vs. 28.33 ± 13.72 months, p = 0.038) and worse functional impairment (MRC sum score, 46.08 ± 12.69 vs. 52.81 ± 7.34, p = 0.018; mRS, 2.76 ± 0.93 vs. 2.33 ± 0.99, p = 0.082; INCAT, 3.68 ± 1.76 vs. 3.03 ± 2.28, p = 0.056, respectively). Electrophysiological studies further revealed greater axonal impairment (4.15 ± 2.0 vs. 5.94 ± 2.77 mv, p = 0.011, ulnar CMAP) and more severe demyelination (5.56 ± 2.86 vs. 4.18 ± 3.71 ms, p = 0.008, ulnar distal latency, 7.94 ± 5.62 vs. 6.52 ± 6.64 ms, p = 0.035, median distal latency; 30.21 ± 12.59 vs. 37.48 ± 12.44 m/s, p = 0.035, median conduction velocity; 58.66 ± 25.73 vs. 42.30 ± 13.77 ms, p = 0.033, median F-wave latency), compared to non-refractory CIDP. Disease duration was shown to be an independent risk factor for refractory CIDP (p < 0.05, 95%CI [0.007, 0.076]). Conclusion: This study provided a comprehensive description of refractory CIDP, addressing its clinical features, classification of clinical course, electrophysiological characteristics, and prognostic factors, effectively elucidating its various aspects. These findings contribute to a better understanding of this challenging subset of CIDP and might be informative for management and treatment strategies.

Non-coding CGG repeat expansion in LOC642361/NUTM2B-AS1 is associated with a phenotype of oculopharyngodistal myopathy.

BACKGROUND: Oculopharyngodistal myopathy (OPDM) is a rare adult-onset neuromuscular disease, associated with CGG repeat expansions in the 5' untranslated region of LRP12, GIPC1, NOTCH2NLC and RILPL1. However, the genetic cause of a proportion of pathoclinically confirmed cases remains unknown. METHODS: A total of 26 OPDM patients with unknown genetic cause(s) from 4 tertiary referral hospitals were included in this study. Clinical data and laboratory findings were collected. Muscle samples were observed by histological and immunofluorescent staining. Long-read sequencing was initially conducted in six patients with OPDM. Repeat-primed PCR was used to screen the CGG repeat expansions in LOC642361/NUTM2B-AS1 in all 26 patients. RESULTS: We identified CGG repeat expansion in the non-coding transcripts of LOC642361/NUTM2B-AS1 in another two unrelated Chinese cases with typical pathoclinical features of OPDM. The repeat expansion was more than 70 times in the patients but less than 40 times in the normal controls. Both patients showed no leucoencephalopathy but one showed mild cognitive impairment detected by Montreal Cognitive Assessment. Rimmed vacuoles and p62-positive intranuclear inclusions (INIs) were identified in muscle pathology, and colocalisation of CGG RNA foci with p62 was also found in the INIs of patient-derived fibroblasts. CONCLUSIONS: We identified another two unrelated cases with CGG repeat expansion in the long non-coding RNA of the LOC642361/NUTM2B-AS1 gene, presenting with a phenotype of OPDM. Our cases broadened the recognised phenotypic spectrum and pathogenesis in the disease associated with CGG repeat expansion in LOC642361/NUTM2B-AS1.

Eomesodermin expression in CD4+T-cells associated with disease progression in amyotrophic lateral sclerosis.

AIM: To clarify the role of Eomesodermin (EOMES) to serve as a disease-relevant biomarker and the intracellular molecules underlying the immunophenotype shifting of CD4+T subsets in amyotrophic lateral sclerosis (ALS). METHODS: The derivation and validation cohorts included a total of 148 ALS patients and 101 healthy controls (HCs). Clinical data and peripheral blood were collected. T-cell subsets and the EOMES expression were quantified using multicolor flow cytometry. Serum neurofilament light chain (NFL) was measured. In 1-year longitudinal follow-ups, the ALSFRS-R scores and primary endpoint events were further recorded in the ALS patients of the validation cohort. RESULTS: In the derivation cohort, the CD4+EOMES+T-cell subsets were significantly increased (p < 0.001). EOMES+ subset was positively correlated with increased serum NFL levels in patients with onset longer than 12 months. In the validation cohort, the elevated CD4+EOMES+T-cell proportions and their association with NFL levels were also identified. The longitudinal study revealed that ALS patients with higher EOMES expression were associated with higher progression rates (p = .010) and worse prognosis (p = .003). CONCLUSIONS: We demonstrated that increased CD4+EOMES+T-cell subsets in ALS were associated with disease progression and poor prognosis. Identifying these associations may contribute to a better understanding of the immunopathological mechanism of ALS.

Electrostatic Self-Assembled Synthesis of Amorphous/Crystalline g-C3N4 Homo-Junction for Efficient Photocatalytic H2 Production with Simultaneous Antibiotic Degradation.

g-C3N4 has been regarded as a promising photocatalyst for photo-reforming antibiotics for H2 production but still suffers from its high charge recombination, which has been proven to be solvable by constructing a g-C3N4 homo-junction. However, those reported methods based on uncontrollable calcination for preparing a g-C3N4 homo-junction are difficult to reproduce. Herein, an amorphous/crystalline g-C3N4 homo-junction (ACN/CCN) was successfully synthesized via the electrostatic self-assembly attachment of negatively charged crystalline g-C3N4 nanorods (CCN) on positively charged amorphous g-C3N4 sheets (ACN). All the ACN/CCN samples displayed much higher photo-reforming of antibiotics for H2 production ability than that of pristine ACN and CCN. In particular, ACN/CCN-2 with the optimal ratio exhibited the best photocatalytic performance, with a H2 evolution rate of 162.5 µmol·g-1·h-1 and simultaneous consecutive ciprofloxacin (CIP) degradation under light irradiation for 4 h. The UV-vis diffuse reflectance spectra (DRS), photoluminescence (PL), and electrochemical results revealed that a homo-junction is formed in ACN/CCN due to the difference in the band arrangement of ACN and CCN, which effectively suppressed the charge recombination and then led to those above significantly enhanced photocatalytic activity. Moreover, H2 was generated from the water reduction reaction with a photogenerated electron (e-), and CIP was degraded via a photogenerated hole (h+). ACN/CCN exhibited adequate photostability and reusability for photocatalytic H2 production with simultaneous CIP degradation. This work provides a new idea for rationally designing and preparing homo-junction photocatalysts to achieve the dual purpose of chemical energy production and environmental treatment.

Construction of a CRISPR Interference System for Gene Knockdown in Stenotrophomonas maltophilia AGS-1 from Aerobic Granular Sludge.

To identify the function of attachment genes involved in biofilm formation in Stenotrophomonas maltophilia AGS-1 isolated from aerobic granular sludge, an effective gene molecular tool is needed. We developed a two-plasmid CRISPRi system in Stenotrophomonas maltophilia AGS-1. One plasmid expressed dCas9 protein with the l-arabinose inducible promoter, and the other plasmid contained the sgRNA cassette complementary to the target gene. Under control of the araC-inducible promoter, this system exhibited little leaky basal expression and highly induced expression that silenced endogenous and exogenous genes with reversible knockdown. This system achieved up to 211-fold suppression for mCherry expression on the nontemplate strand compared to the template strand (91-fold). The utility of the developed CRISPRi platform was also characterized by suppressing the xanA and rpfF genes. The expression of these two genes was rapidly depleted and the adhesion ability decreased, which demonstrated that the modulation of either gene was an important factor for biofilm formation of the AGS-1 strain. The system also tested the ability to simultaneously silence transcriptional suppression of multiple targeted genes, an entire operon, or part of it. Lastly, the use of CRISPRi allowed us to dissect the gene intricacies involved in flagellar biosynthesis. Collectively, these results demonstrated that the CRISPRi system was a simple, feasible, and controllable manipulation system of gene expression in the AGS-1 strain.

Portal Selection for Suture Anchor Placement During Hip Arthroscopic Labral Repair: A Study Based on 3-Dimensional Model Reconstruction.

Background: Arthroscopic suture repair is the main treatment option for hip labral tears; however, anchor insertion and placement from arthroscopic portals is difficult. Purpose: To quantitatively evaluate the safety of various arthroscopic portals for suture anchor placement during hip labral repair. Study Design: Descriptive laboratory study. Methods: The computed tomography scans of 20 patients with normally developed hip joints were used to create 3-dimensional models. The distances from the anchor to the articular cartilage (DAC) and from the acetabular insertion point to the cortical bone (DCB) were measured in the anterolateral portal (AL), posterolateral portal (PL), midanterior portal (MAP), medial MAP, and 3 distal anterolateral accessory portals (DALAs): DALA-proximal, DALA-middle, and DALA-distal. Labral tears were divided into anterior (4, 3, and 2 o'clock), lateral (1, 12, and 11 o'clock), and posterior (10, 9, and 8 o'clock) acetabular zones, and the Kruskal-Wallis and Mann-Whitney U test were used to compare DAC and DCB in the zones. The success rate was defined as anchors placed with DAC ≥1 mm and DCB ≥15 mm. Results: The DAC was significantly smaller in the AL at 1 o'clock (0.68 ± 0.32 mm; P < .001) and 12 o'clock (0.37 ± 0.30 mm; P < .001), and in the PL at 12 o'clock (-0.35 ± 0.38 mm; P < .001) and 11 o'clock (0.60 ± 0.24 mm; P < .001). The DCB was significantly smaller in the DALA-P at 3 o'clock (8.93 ± 2.12 mm; P < .001) and 11 o'clock (9.59 ± 2.84 mm; P < .001), the MAP at 12 o'clock (13.76 ± 3.89 mm; P < .001) and 11 o'clock (0.27 ± 0.27 mm; P < .001), and the MMA at 12 o'clock (5.96 ± 2.31 mm; P < .001) and 11 o'clock (0 mm; P < .001). Success rates were high for MAP and MMA between 4 o'clock and 1 o'clock, for DALA-proximal at 12 o'clock, for AL at 11 o'clock, and for PL between 10 o'clock and 8-o'clock. Conclusion: There were significant differences in the success rate of anchor placement using different portals during hip arthroscopic labral repair. Clinical Relevance: MAP is recommended for labral repair between 4 o'clock and 1 o'clock, DALA-P is recommended between 2 o'clock and 12 o'clock, AL is suitable at 11 o'clock, and PL is suitable between 10 o'clock and 8 o'clock.

Hyperpolarized 13C metabolic imaging detects long-lasting metabolic alterations following mild repetitive traumatic brain injury.

Career athletes, active military, and head trauma victims are at increased risk for mild repetitive traumatic brain injury (rTBI), a condition that contributes to the development of epilepsy and neurodegenerative diseases. Standard clinical imaging fails to identify rTBI-induced lesions, and novel non-invasive methods are needed. Here, we evaluated if hyperpolarized 13C magnetic resonance spectroscopic imaging (HP 13C MRSI) could detect long-lasting changes in brain metabolism 3.5 months post-injury in a rTBI mouse model. Our results show that this metabolic imaging approach can detect changes in cortical metabolism at that timepoint, whereas multimodal MR imaging did not detect any structural or contrast alterations. Using Machine Learning, we further show that HP 13C MRSI parameters can help classify rTBI vs. Sham and predict long-term rTBI-induced behavioral outcomes. Altogether, our study demonstrates the potential of metabolic imaging to improve detection, classification and outcome prediction of previously undetected rTBI.

Construction and application of a mCherry fluorescent labeling system for Stenotrophomonas AGS-1 from aerobic granular sludge.

To elucidate the specific mechanism by which high-attachment bacteria promote aerobic granular sludge (AGS) formation, a red fluorescent protein mCherry-based biomarker system was developed in the high-attachment strain Stenotrophomonas AGS-1 from AGS. The fluorescent labeling system used plasmid-mediated mCherry expression driven by a Ptac constitutive promoter. mCherry-labeled AGS-1 had normal unimpaired growth, strong fluorescent signals, and good fluorescence imaging. Also, the mCherry labeling system had no effect on the attachment ability of AGS-1. In addition, mCherry-labeled AGS-1 maintained high plasmid stability, even after more than 100 generations. Notably, after the addition of mCherry-labeled AGS-1 into the activated sludge system, the mCherry fluorescence of the sludge system can be used as a good reflection of the relative amount of AGS-1. Moreover, the spatial distribution of mCherry-labeled AGS-1 in the sludge system could be visualized and remained clear even after 5 days by fluorescence imaging. These results revealed that the mCherry-based biomarker system would provide a valuable tool for labeling AGS-1 to monitor the spatial distribution and fate of AGS-1 in AGS, which would help to better understand the mechanism of AGS formation and facilitate the development of AGS technology.

Intrafamilial phenotypic heterogeneity in GIPC1-related oculopharyngodistal myopathy type 2: a case report.

Oculopharyngodistal myopathy (OPDM) is a rare adult-onset neuromuscular disease characterized by ocular, facial, bulbar and distal limb muscle weakness. Here, we presented a pair of siblings with OPDM2 displaying marked intrafamilial phenotypic heterogeneity. In addition to muscle weakness, the proband also demonstrated tremor and visual disturbance that have not been reported previously in OPDM2. Electrophysiological and pathological studies further suggested the presence of neurogenic impairment in the proband. Repeat-primed polymerase chain reaction (RP-PCR) and fluorescence amplicon length analysis polymerase chain reaction (AL-PCR) confirmed the molecular diagnosis of OPDM2 in the siblings. Given the rarity of the case, the association between OPDM2 and tremor, visual disturbance, or neurogenic impairment remained to be explored.

Construction and application of a new CRISPR/Cas12a system in Stenotrophomonas AGS-1 from aerobic granular sludge.

Aerobic granular sludge (AGS) is a microbial aggregate with a biofilm structure. Thus, investigating AGS in the aspect of biofilm and microbial attachment at the genetic level would help to reveal the mechanism of granule biofilm formation. In this work, a two-plasmid clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated proteins (Cas)12a genome editing system was constructed to identify attachment genes for the first time in Stenotrophomonas AGS-1 from AGS. One plasmid contained a Cas12a cassette driven by an arabinose-inducible promoter, and another contained the specific crRNA and homologous arms (HAs). Acidaminococcus sp. Cas12a (AsCas12a) was adopted and proven to have mild toxicity (compared to Cas9) and strong cleavage activity for AGS-1. CRISPR/Cas12a-mediated rmlA knockout decreased attachment ability by 38.26%. Overexpression of rmlA in AGS-1 resulted in an increase of 30.33% in attachment ability. These results showed that the modulation of rmlA was an important factor for the biofilm formation of AGS-1. Moreover, two other genes (xanB and rpfF) were knocked out by CRISPR/Cas12a and identified as attachment-related genes in AGS-1. Also, this system could achieve point mutations. These data indicated that the CRISPR/Cas12a system could be an effective molecular platform for attachment gene function identification, which would be useful for the development of AGS in wastewater treatment.