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Femur fracture leads to loss of bone at uninjured skeletal sites, which may increase risk of subsequent fracture. Osteocytes, the most abundant bone cells, can directly resorb bone matrix and regulate osteoclast and osteoblast activity, but their role in systemic bone loss after fracture remains poorly understood. In this study we used a transgenic (TG+) mouse model that overexpresses human B-cell lymphoma 2 (BCL-2) in osteoblasts and osteocytes. This causes enhanced osteoblast proliferation, followed by disruption in lacunar-canalicular connectivity and massive osteocyte death by 10 wk of age. We hypothesized that reduced viable osteocyte density would decrease the magnitude of systemic bone loss after femur fracture, reduce perilacunar remodeling, and alter callus formation. Bone remodeling was assessed using serum biomarkers of bone formation and resorption at 5 d post-fracture. We used micro-computed tomography, high resolution x-ray microscopy, mechanical testing, and Raman spectroscopy to quantify the magnitude of systemic bone loss, as well as changes in osteocyte lacunar volume, bone strength, and bone composition 2 wk post-fracture. Fracture was associated with a reduction in circulating markers of bone resorption in non-transgenic (TG-) animals. TG+ mice exhibited high bone mass in the limbs, greater cortical elastic modulus and reduced post-yield displacement. After fracture, TG+ mice lost less trabecular bone than TG- mice, but conversely TG+ mice exhibited trends toward a lower yield point and reduced femoral cortical thickness after fracture, though these were not statistically significant. Lacunar density was greater in TG+ mice, but fracture did not alter lacunar volume in TG+ or TG- mice. These findings suggest that osteocytes potentially play a significant role in the post-traumatic systemic response to fracture, though the effects differ between trabecular and cortical bone.

Post-traumatic osteoarthritis (PTOA) occursfollowing an injury such as Anterior CruciateLigament (ACL) rupture. Clinically, ACLreconstruction surgery is performedfollowing injury. However, it is unclear howeffective knee restabilization is in slowingPTOA progression. This study hypothesized that knee restabilization would slow the progression of PTOA, but the procedure itself would have some mild PTOA-like joint degeneration.

Noninvasive compression-induced anterior cruciate ligament rupture (ACL-R) is an easy and reproducible model for studying post-traumatic osteoarthritis (PTOA) in mice. However, equipment typically used for ACL-R is expensive, immobile, and not available to all researchers. In this study, we compared PTOA progression in mice injured with a low-cost custom ACL-rupture device (CARD) to mice injured with a standard system (ElectroForce 3200). We quantified anterior-posterior (AP) joint laxity immediately following injury, epiphyseal trabecular bone microstructure, and osteophyte volume at 2 and 6 weeks post injury using micro-computed tomography, and osteoarthritis progression and synovitis at 2 and 6 weeks post injury using whole-joint histology. We observed no significant differences in outcomes in mice injured with the CARD system compared to mice injured with the Electroforce (ELF) system. However, AP joint laxity data and week 2 micro-CT and histology outcomes suggested that injuries may have been slightly more severe and PTOA progressed slightly faster in mice injured with the CARD system compared to the ELF system. Altogether, these data confirm that ACL-R can be successfully and reproducibly performed with the CARD system and that osteoarthritis (OA) progression is mostly comparable to that of mice injured with the ELF system, though potentially slightly faster. The CARD system is low cost and portable, and we are making the plans and instructions freely available to all interested investigators in the hopes that they will find this system useful for their studies of OA in mice.

Traumatic joint injuries such as anterior cruciate ligament (ACL) rupture or meniscus tears commonly lead to post-traumatic osteoarthritis (PTOA) within 10-20 years following injury. Understanding the early biological processes initiated by joint injuries (e.g., inflammation, matrix metalloproteinases (MMPs), cathepsin proteases, bone resorption) is crucial for understanding the etiology of PTOA. However, there are few options for in vivo measurement of these biological processes, and the early biological responses may be confounded if invasive surgical techniques or injections are used to initiate OA. In our studies of PTOA, we have used commercially available near-infrared protease activatable probes combined with fluorescence reflectance imaging (FRI) to quantify protease activity in vivo following non-invasive compression-induced ACL injury in mice. This non-invasive ACL injury method closely recapitulates clinically relevant injury conditions and is completely aseptic since it does not involve disrupting the skin or the joint capsule. The combination of these injury and imaging methods allows us to study the time course of protease activity at multiple time points following a traumatic joint injury.

While adult fireflies are terrestrial, their larvae inhabit various habitat types, and a lack of comprehensive research on the feeding habits of these larvae across different habitats has greatly impeded the development of artificial diets. Here, we tested 14 types of foods, primarily covering gastropods, vertebrates, and fruit, to survey feed for aquatic (Aquatica leii and Sclerotia substriata), semi-aquatic (Pygoluciola qingyu and Pygoluciola sp.), and terrestrial (Pyrocoelia analis) fireflies. The results show that A. leii, S. substriata, P. qingyu, Pygoluciola sp., and P. analis fed on 12, 6, 10, 10, and 7 different foods, respectively, showing an obvious difference in feeding range among various inhabit types of fireflies. Aquatic lineages preferred to consume freshwater snails, followed by pork meat and land slugs, while semi-aquatic fireflies favored freshwater snails, followed by fish and shrimp. Favorite foods were thus freshwater snails such as Cipangopaludina chinensis for both aquatic and semi-aquatic fireflies, but feeding preference differed for second favorite foods (e.g., pork vs. fish meat). Terrestrial Pyrocoelia analis showed different feeding preferences compared withthe other two habitat lineages, with terrestrial snails and slugs as their favorite foods, followed by freshwater snails, such as C. chinensis. These findings not only uncovered larval feeding habits of fireflies across various inhabit types but also indicated that readily available and affordable C. chinensis products can serve as wild snail alternatives in the artificial feeding of fireflies. This study is the first to explore the feeding habits of firefly species inhabiting water and land habitat types, adding to the understanding of the feeding characteristics of fireflies.

We previously showed that femur fracture in mice caused a reduction in bone volume at distant skeletal sites within 2 weeks post-fracture. Osteocytes also have the ability to remodel their surrounding bone matrix through perilacunar/canalicular remodeling (PLR). If PLR is altered systemically following fracture, this could affect bone mechanical properties and increase fracture risk at all skeletal sites. In this study, we investigated whether lacunar-canalicular microstructure and the rate of PLR are altered in the contralateral limb following femoral fracture in mice. We hypothesized that femoral fracture would accelerate PLR by 2 weeks postfracture, followed by partial recovery by 4 weeks. We used histological evaluation and high-resolution microcomputed tomography to quantify the morphology of the lacunar-canalicular network at the contralateral tibia, and we used quantitative real-time polymerase chain reaction (RT-PCR) and RNA-seq to measure the expression of PLR-associated genes in the contralateral femur. We found that at both 2 and 4 weeks postfracture, canalicular width was significantly increased by 18.6% and 16.6%, respectively, in fractured mice relative to unfractured controls. At 3 days and 4 weeks post-fracture, we observed downregulation of PLR-associated genes; RNA-seq analysis at 3 days post-fracture showed a deceleration of bone formation and mineralization in the contralateral limb. These data demonstrate notable canalicular changes following fracture that could affect bone mechanical properties. These findings expand our understanding of systemic effects of fracture and how biological and structural changes at distant skeletal sites may contribute to increased fracture risk following an acute injury.

ABSTRACT: Phenyllactic acid (PLA) has been demonstrated to possess antibacterial activity and capacity to prolong food shelf life. However, studies on the performance of PLA in inhibiting Staphylococcus aureus and its effectiveness when applied to dairy products are largely lacking. Here, antibacterial activity (planktonic and biofilm states) of PLA against S. aureus CICC10145 (S. aureus_45) were investigated. The results showed that PLA inhibited growth of S. aureus_45 and formation of S. aureus_45 biofilm. Next, the antibacterial action target of PLA was uncovered from both physiological and phenotypic perspectives. The results showed that PLA decreased cell metabolic activity and cell viability, damaged cell membrane integrity, triggered leakage of intracellular contents (DNA, proteins, and ATP), and caused oxidative stress damage and morphological deformation of S. aureus_45. In practical application, the antibacterial activity of PLA against S. aureus_45 cells was further confirmed in skim milk and cheese as dairy food models, and the antibacterial effects can be adequately maintained during storage for 21 d, at least at 4°C. These findings suggested that PLA could be a potential candidate for controlling S. aureus outgrowth in dairy foods.

Summary: Knowledge about the impact of different dissolved oxygen (DO) on the composition and function of gut bacteria of aquatic insects is largely unknown. Herein, we constructed freshwater environments with different DOs (hypoxia: 2.50 ± 0.50, normoxia: 7.00 ± 0.50, and hyperoxia: 13.00 ± 0.50 mg/L) where aquatic firefly Aquatica leii larvae lived for three months. Their gut flora was analyzed using the combination of 16S rRNA amplicon sequencing and metagenomics. The results showed no difference in alpha diversity of the gut flora between A. leii inhabiting various DOs. However, the relative abundance of several bacterial lineages presented significant changes, such as Pseudomonas. In addition, bacterial genes with an altered relative abundance in response to various DOs were primarily related to metabolism. The alteration of these functions correlated with the DO change. This is the first to uncover structure of gut flora under various DOs in aquatic insect larvae.

ABSTRACT: Staphylococcus aureus and its biofilm have emerged as a significant threat to the safety of dairy products. In recent years, lactic acid bacteria (LAB) bacteriocins have been widely acknowledged as the potential natural antibacterial substance in food biopreservation due to their excellent antibacterial effects. However, few LAB bacteriocins with antibacterial and antibiofilm activity against S. aureus have been reported in dairy products. In the present study, a novel bacteriocin LSX01 of Lactobacillus paracasei LS-6 isolated from a traditional fermented yogurt produced in Yunnan, China, was purified and characterized extensively. The LSX01 possessed a molecular weight of 967.49 Da and an AA sequence of LDQAGISYT. The minimum inhibitory concentration of LSX01 against S. aureus_45 was 16.90 μg/mL, which was close to or lower than the previously reported bacteriocins. The LSX01 exhibited an extensive antimicrobial spectrum against both gram-positive and gram-negative bacteria. Moreover, LSX01 exhibited excellent tolerance to heat and acid-base treatments, and sensitivity to the proteolytic enzymes, such as pepsin and proteinase K. Furthermore, the treatment of S. aureus_45 planktonic cells with LSX01 significantly reduced their metabolic activity and disrupted the cell membrane integrity. Scan electron microscopy results demonstrated that LSX01 induced cytoplasmic content leakage and cell deformation. Additionally, biofilm formation of S. aureus_45 was also significantly inhibited by LSX01. Overall, the results suggested that the novel LAB bacteriocin LSX01 possessed antibacterial activity and antibiofilm activity against S. aureus and, hence, could have potential for improving safety of dairy products.

Mitochondria are the leading organelle for energy metabolism. The toxic effects of environmental toxicants on mitochondrial morphology, energy metabolism, and their determination of cell fate have already been broadly studied. However, minimal research exists on effects of environmental toxicants such as pesticides on mitochondrial energy metabolism at in vitro subcellular level, particularly from an omics perspectives (e.g., metabolomics). Here, German cockroach (Blattella germanica) was fed diets with (0.01 and 0.001 mg/mL) and without abamectin, and highly purified fat body mitochondria were isolated. Swelling measurement confirmed abnormal mitochondrial swelling caused by abamectin stress. The activity of two key mitochondrial energy metabolism-related enzymes, namely succinic dehydrogenase and isocitrate dehydrogenase, was significantly affected. The metabolomic responses of the isolated mitochondria to abamectin were analyzed via untargeted liquid chromatography/mass spectrometry metabolomics technology. Fifty-two differential metabolites (DMs) were identified in the mitochondria between the 0.001 mg/mL abamectin-fed and the control groups. Many of these DMs were significantly enriched in pathways involved in ATP production and energy consumption (e.g., oxidative phosphorylation, TCA cycle, and pentose phosphate pathway). Nineteen of the DMs were typically related to energy metabolism. This study is valuable for further understanding mitochondrial toxicology under environmental toxicants, particularly its subcellular level.

Osteopetrosis (OS) is a rare heritable disorder characterized by osteoclast dysfunction and increased bone density on radiography. Optic nerve osseous compression is the most frequent ocular complication of OS, with nystagmus, strabismus, ptosis, proptosis, and lagophthalmos occurring less frequently. However, it is uncommon for patients to have neurological or ocular symptoms at initial presentation. Herein, we present the case of a 3-year-old girl with the initial presentation of ocular symptoms who was confirmed to have OS through genetic testing. She was born full-term and found to have nystagmus since the age of 1 year. Her best-corrected visual acuity was 1.2/60 for both eyes. Exotropia of the left eye and bilateral small-amplitude pendular nystagmus were also noted. Color fundoscopy revealed a tessellated fundus and pale discs with cup-to-disc ratios of 0.5–0.6. Magnetic resonance imaging revealed bilateral optic canal stenosis and optic nerve atrophy. Whole-exome sequencing revealed a biallelic chloride voltage-gated channel 7 mutation, c.2297T > C (p.Leu766Pro) and c.1577G > A (p.Arg526Gln), and autosomal recessive OS was diagnosed. The patient is currently being evaluated for possible hematopoietic stem cell transplantation. We suggest that OS should be considered a differential diagnosis for unexplained nystagmus and optic nerve atrophy. [ABSTRACT FROM AUTHOR]

Underwater images often encounter challenges such as attenuation, color distortion, and noise caused by artificial lighting sources. These imperfections not only degrade image quality but also impose constraints on related application tasks. Improving underwater image quality is crucial for underwater activities. However, obtaining clear underwater images has been a challenge, because scattering and blur hinder the rendering of true underwater colors, affecting the accuracy of underwater exploration. Therefore, this paper proposes a new deep network model for single underwater image enhancement. More specifically, our framework includes a light field module (LFM) and sketch module, aiming at the generation of a light field map of the target image for improving the color representation and preserving the details of the original image by providing contour information. The restored underwater image is gradually enhanced, guided by the light field map. The experimental results show the better image restoration effectiveness, both quantitatively and qualitatively, of the proposed method with a lower (or comparable) computing cost, compared with the state-of-the-art approaches. [ABSTRACT FROM AUTHOR]

The applications of image classification are useful, especially in the prediction of object categories at a high accuracy rate. The technique helps industrial designers classify the existing products by product appearance and assess the extent to which newly designed products belong to their brand or product family. By doing this, designers classify products by typicality. Yet, the application of image classification of products and brand styles based on typicality is still limited. Thus, we presented a case study in which the extent to which a chair was recognized as a typical Apple product was predicted. In the technique, Waikato Environment for Knowledge Analysis (WEKA) was used. The results show that the industrial designer can use product typicality for the image classification of products to manage the style of a brand and its products. [ABSTRACT FROM AUTHOR]

Patients with refractory diabetes are defined as type 2 diabetes (T2D) patients; they cannot achieve optimal glycemic control and exhibit persistent elevations of hemoglobin A1c (HbA1c) ≥8% while on appropriate therapy. Hyperglycemia can lead to severe microvascular/macrovascular complications. However, in contrast to T2D, few studies have focused specifically on the gut microbiota in refractory diabetes. To examine this issue, we recruited 79 subjects with T2D and refractory diabetes (RT2D), and all subjects received standard therapy with Metformin or other hypoglycemic agents with or without insulin for at least one year. The α-diversity displayed no significant difference, whereas the β-diversity showed a marginal significance (p = 0.054) between T2D and RT2D. The evaluation of taxonomic indices revealed reductions in both Akkermansia muciniphila and Fusobacterium and a corresponding enrichment of Bacteroides vulgatus, Veillonella denticariosi among those with RT2D. These microbial markers distinguished RT2D from T2D with an acceptable degree of discrimination (area under the curve (AUC) = 0.719, p < 0.01) and were involved in several glucose-related functional pathways. Furthermore, the relative abundance of Akkermansia muciniphila was negatively correlated with HbA1c. Our combined results reveal unique features of the gut microbiota in RT2D and suggest that the evaluation of the gut microbiota could provide insights into the mechanisms underlying glycemic control and the impact of therapeutic modalities in patients with RT2D.

Horner Syndrome (HS) is characterized by symptoms of ipsilateral miosis, ptosis, enophthalmos, and facial anhidrosis, which is caused by the damaged oculosympathetic pathway. HS is rarely reported as postoperative complications of fine‐needle aspiration (FNA). We report a case of HS triggered by Ultrasound‐guided FNA during thyroid cancer management and conducted the literature review. A 31‐year‐old male with differentiated thyroid cancer underwent total thyroidectomy and regional lymph node dissection as well as radioactive iodine ablation, presented with persistently elevated tumor marker of thyroglobulin and suspicious left level IV and V cervical lymph nodes by neck ultrasound. Ultrasound‐guided left cervical lymph nodes FNA for cellular diagnosis was performed, and typical manifestations of HS appeared immediately after the procedure. Subsequent ultrasound evaluation of the same area demonstrated a subtle strip of the hypo‐echogenic area in the superior pole of the suspected level IV structure, suggesting sympathetic ganglia with the visible originating nerve fiber on the superior pole. All of the patient's symptoms of HS were resolved 2 months after the incidence. Cervical sympathetic ganglia can be similar in size, shape, and ultrasound characteristics to a malignant lymph node. Thorough ultrasound examination by directly comparing the potential ganglia with a typical malignant lymph node, and paying attention to any potential root fibers on the target is key to avoiding ganglia injury before the neck invasive procedures. [ABSTRACT FROM AUTHOR]