bluuebunny/crossref_metadata_2025 huggingface dataset:
Characterization of feline-originated probiotics Lactobacillus rhamnosus CACC612 and Bifidobacterium animalis subsp. lactis CACC789 and evaluation of their host response
Authors: Jang Hyun-Jun , Kim Jung-Ae , Kim Yangseon
Published: April 2024

Background

Probiotics are beneficial for animal health and new potential probiotics need to be characterized for their prospective use in improving animal health. In this study, 32 bacterial strains were isolated from a Norwegian forest cat (castrated, 12 years old) and a Persian cat (castrated, 10 years old), which were privately owned and had indoor access.

Results

Lactobacillus rhamnosus CACC612 (CACC612) and Bifidobacterium animalis subsp. lactis CACC789 (CACC789) were selected as potential probiotics; characterization of the two strains showed equivalent acid tolerance, similar cell adhesion rates on the HT-29 monolayer cell line, and superior bile tolerance compared to Lactobacillus rhamnosus GG (LGG). Subsequently, they exhibited inhibitory effects against a broad spectrum of pathogenic bacteria, including E. coli (KCTC 2617), Salmonella Derby (NCCP 12,238), Salmonella Enteritidis (NCCP 14,546), Salmonella Typhimurium (NCCP 10,328), Clostridium difficile JCM 1296T. From evaluating host effects, the viability of the feline macrophage cell line (Fcwf-4) increased with the treatment of CACC612 or CACC789 (P < 0.05). The induced expression of immune-related genes such as IFN-γ, IL1β, IL2, IL4, and TNF-α by immune stimulation was significantly attenuated by the treatment of CACC612 or CACC789 (P < 0.05). When 52 clinical factors of sera from 21 healthy cats were analyzed using partial least squares discriminant analysis (PLS-DA), the animals were obviously clustered before and after feeding with CACC612 or CACC789. In addition, hemoglobin and mean corpuscular hemoglobin concentration (MCHC) significantly increased after CACC612 feeding (P < 0.05).

Conclusions

In this study, feline-originated probiotics were newly characterized and their potentially probiotic effects were evaluated. These results contribute to our understanding of the functional effects of feline-derived probiotics and support their industrial applications.
Prevalence, genetic diversity, and antimicrobial susceptibility of Vibrio spp. infected gilthead sea breams from coastal farms at Damietta, Egypt
Authors: Ismail Esraa Tawfeek , El-Son Mai A. M. , El-Gohary Fatma A. , Zahran Eman
Published: April 2024

Background

Vibriosis is one of the most serious bacterial diseases and causes high morbidity and mortality among cultured sea breams. This study was undertaken to track the surveillance of Vibrio infection and its correlation to environmental factors. A total of 115 gilthead sea breams were collected seasonally from a private earthen pond fish farm in the Shatta area of Damietta, Egypt from September 2022 to July 2023. Physicochemical parameters of water were analyzed, and heavy metal levels were measured. The fish samples were subjected to clinical, bacteriological, Enterobacterial Repetitive Intergenic Consensus (ERIC) fingerprinting, and hematoxylin and Eosin histopathological staining.

Results

The results revealed significant variations in the water quality parameters over different seasons, in addition to an increase in heavy metals. Naturally infected fish showed external signs and postmortem lesions that were relevant to bacterial infection. Two dominant Vibrio subspecies of bacteria were identified: V. alginolyticus (205 isolates) and V. fluvialis (87 isolates). PCR confirmed the presence of V. alginolyticus using the species-specific primer collagenase at 737 bp. The highest prevalence of V. alginolyticus was detected during the summer season (57.72%), and the lowest prevalence was observed in autumn (39.75%). The correlation analysis revealed a positive relationship between V. alginolyticus and water temperature (r = 0.69). On the other hand, V. fluvialis showed a high prevalence during the autumn season (25.30%) and the lowest prevalence during the summer season (10.56%), where it was negatively correlated with water temperatures (r =—0.03). ERIC fingerprinting showed genetic variation within the Vibrio isolates. Antimicrobial susceptibility testing revealed sensitivity to ciprofloxacin and doxycycline, and resistance to amoxicillin and erythromycin. The multiple antibiotic resistance (MAR) index values for V. alginolyticus and V. fluvialis ranged from 0.3 to 0.7, with a multi-drug resistance pattern to at least three antibiotics. Histopathological alterations in the affected tissues revealed marked hemorrhage, vascular congestion, and hemosiderosis infiltration.

Conclusion

This study provides insights into the potential propagation of waterborne diseases and antibiotic resistance in the environment. Ensuring that the environment does not serve as a reservoir for virulent and contagious Vibrio species is a critical concern for regional aquaculture industries. Therefore, we recommend implementing environmental context-specific monitoring and surveillance tools for microbial resistance.
Transcriptomics aids in uncovering the metabolic shifts and molecular machinery of Schizochytrium limacinum during biotransformation of hydrophobic substrates to docosahexaenoic acid
Authors: Mariam Iqra , Krikigianni Eleni , Rantzos Chloe , Bettiga Maurizio , Christakopoulos Paul , Rova Ulrika , Matsakas Leonidas , Patel Alok
Published: April 2024

Background

Biotransformation of waste oil into value-added nutraceuticals provides a sustainable strategy. Thraustochytrids are heterotrophic marine protists and promising producers of omega (ω) fatty acids. Although the metabolic routes for the assimilation of hydrophilic carbon substrates such as glucose are known for these microbes, the mechanisms employed for the conversion of hydrophobic substrates are not well established. Here, thraustochytridSchizochytrium limacinumSR21 was investigated for its ability to convert oils (commercial oils with varying fatty acid composition and waste cooking oil) into ω-3 fatty acid; docosahexaenoic acid (DHA).

Results

Within 72 h SR21 consumed ~ 90% of the oils resulting in enhanced biomass (7.5 g L− 1) which was 2-fold higher as compared to glucose. Statistical analysis highlights C16 fatty acids as important precursors of DHA biosynthesis. Transcriptomic data indicated the upregulation of multiple lipases, predicted to possess signal peptides for secretory, membrane-anchored and cytoplasmic localization. Additionally, transcripts encoding for mitochondrial and peroxisomal β-oxidation along with acyl-carnitine transporters were abundant for oil substrates that allowed complete degradation of fatty acids to acetyl CoA. Further, low levels of oxidative biomarkers (H2O2, malondialdehyde) and antioxidants were determined for hydrophobic substrates, suggesting that SR21 efficiently mitigates the metabolic load and diverts the acetyl CoA towards energy generation and DHA accumulation.

Conclusions

The findings of this study contribute to uncovering the route of assimilation of oil substrates by SR21. The thraustochytrid employs an intricate crosstalk among the extracellular and intracellular molecular machinery favoring energy generation. The conversion of hydrophobic substrates to DHA can be further improved using synthetic biology tools, thereby providing a unique platform for the sustainable recycling of waste oil substrates.
Motivation and context of concurrent stimulant and opioid use among persons who use drugs in the rural United States: a multi-site qualitative inquiry
Authors: Fredericksen R. J. , Baker R. , Sibley A. , Estadt A. T. , Colston D. , Mixson L. S. , Walters S. , Bresett J. , Levander X. A. , Leichtling G. , Davy-Mendez T. , Powell M. , Stopka T. J. , Pho M. , Feinberg J. , Ezell J. , Zule W. , Seal D. , Cooper H. L. F. , Whitney B. M. , Delaney J. A. C. , Crane H. M. , Tsui J. I.
Published: April 2024

Background

In recent years, stimulant use has increased among persons who use opioids in the rural U.S., leading to high rates of overdose and death. We sought to understand motivations and contexts for stimulant use among persons who use opioids in a large, geographically diverse sample of persons who use drugs (PWUD) in the rural settings.

Methods

We conducted semi-structured individual interviews with PWUD at 8 U.S. sites spanning 10 states and 65 counties. Content areas included general substance use, injection drug use, changes in drug use, and harm reduction practices. We used an iterative open-coding process to comprehensively itemize and categorize content shared by participants related to concurrent use.

Results

We interviewed 349 PWUD (64% male, mean age 36). Of those discussing current use of stimulants in the context of opioid use (n = 137, 39%), the stimulant most used was methamphetamine (78%) followed by cocaine/crack (26%). Motivations for co-use included: 1) change in drug markets and cost considerations; 2) recreational goals, e.g., seeking stronger effects after heightened opioid tolerance; 3) practical goals, such as a desire to balance or alleviate the effects of the other drug, including the use of stimulants to avoid/reverse opioid overdose, and/or control symptoms of opioid withdrawal; and 4) functional goals, such as being simultaneously energized and pain-free in order to remain productive for employment.

Conclusion

In a rural U.S. cohort of PWUD, use of both stimulants and opioids was highly prevalent. Reasons for dual use found in the rural context compared to urban studies included changes in drug availability, functional/productivity goals, and the use of methamphetamine to offset opioid overdose. Education efforts and harm reduction services and treatment, such as access to naloxone, fentanyl test strips, and accessible drug treatment for combined opioid and stimulant use, are urgently needed in the rural U.S. to reduce overdose and other adverse outcomes.
Determinants and predictive model of failure of surgical repair of obstetric vesico-vaginal fistula in the Democratic Republic of the Congo
Authors: Paluku Justin Lussy , Aksanti Barthelemy Kasi , Clemmer William C. , Furaha Cathy Mufungizi , Kamabu Eugénie Mukekulu , Kasereka Jonathan M. L. , Kalole Benjamin Kambale , Mukuku Olivier , Tsongo Zacharie Kibendelwa , Wembonyama Stanis Okitotsho , Mpoy Charles Wembonyama , Juakali Jeannot Sihalikyolo
Published: April 2024

Introduction

Surgical repair of obstetric fistula aims to restore the anatomical and functional integrity of the urinary tract, enabling affected women to regain their dignity and quality of life. However, such repairs can end in a failure. The aim of this study is to develop a predictive score to identify factors influencing failure of surgical repair of obstetric vesico-vaginal fistula (FSROVVF) in the Democratic Republic of the Congo.

Methods

This was an analytical cross-sectional study of 318 women with obstetric vesico-vaginal fistula (OVVF) who had undergone surgical management. A bivariate and then a multivariate analysis were performed. Score discrimination was assessed using the ROC curve and C-index, and score calibration using the Hosmer–Lemeshow test.

Results

Surgical repair of OVVF was unsuccessful in 16.98% of cases (54/318). After logistic modeling, six criteria emerged as predictive factors for FSROVVF: the presence of fibrosis (AOR = 5.01; 95% CI:1.73–14.49), the presence of 2 or more fistulas (AOR = 9.04; 95% CI:3.01–27.13), the association of OVVF with another anatomoclinical entity of fistula (AOR = 3.16; 95% CI:1.09–9.13), the fistula size > 3 cm (AOR = 3.65; 95% CI:1.36–9.76), the peri-operative hemorrhage (AOR = 7.01; 95% CI:2.33–21.03), and the post-operative infection (AOR = 178.89; 95% CI:26.09–1226.64). A score ranging from 0 to 13 points was obtained, of which a value ≤ 5 points defines a low risk of FSROVVF, a value between 6 and 8 points defines a moderate risk and value ≥ 9 points corresponds to a high risk of FSROVVF. The area under the ROC curve of the score is 0.925 with a sensitivity of 61.11%, a specificity of 96.59%, a positive predictive value of 78.57% and a negative predictive value of 92.39%.

Conclusion

This study demonstrated that the number of fistulas ≥ 2, fistula size > 3 cm, fibrosis, association of OVVF with other types of fistulas, peri-operative hemorrhage, and post-operative infection are factors predictive of FSROVVF. These six factors are key contributors to the score used to predict FSROVVF. Once validated, this score will inform and enable preoperative counseling regarding the prognosis and the chances of a successful outcome of surgical repair of OVVF.
Ferroptosis contributing to cardiomyocyte injury induced by silica nanoparticles via miR-125b-2-3p/HO-1 signaling
Authors: Li Xueyan , Xu Hailin , Zhao Xinying , Li Yan , Lv Songqing , Zhou Wei , Wang Ji , Sun Zhiwei , Li Yanbo , Guo Caixia
Published: April 2024

Background

Amorphous silica nanoparticles (SiNPs) have been gradually proven to threaten cardiac health, but pathogenesis has not been fully elucidated. Ferroptosis is a newly defined form of programmed cell death that is implicated in myocardial diseases. Nevertheless, its role in the adverse cardiac effects of SiNPs has not been described.

Results

We first reported the induction of cardiomyocyte ferroptosis by SiNPs in both in vivo and in vitro. The sub-chronic exposure to SiNPs through intratracheal instillation aroused myocardial injury, characterized by significant inflammatory infiltration and collagen hyperplasia, accompanied by elevated CK-MB and cTnT activities in serum. Meanwhile, the activation of myocardial ferroptosis by SiNPs was certified by the extensive iron overload, declined FTH1 and FTL, and lipid peroxidation. The correlation analysis among detected indexes hinted ferroptosis was responsible for the SiNPs-aroused myocardial injury. Further, in vitro tests, SiNPs triggered iron overload and lipid peroxidation in cardiomyocytes. Concomitantly, altered expressions of TfR, DMT1, FTH1, and FTL indicated dysregulated iron metabolism of cardiomyocytes upon SiNP stimuli. Also, shrinking mitochondria with ridge fracture and ruptured outer membrane were noticed. To note, the ferroptosis inhibitor Ferrostatin-1 could effectively alleviate SiNPs-induced iron overload, lipid peroxidation, and myocardial cytotoxicity. More importantly, the mechanistic investigations revealed miR-125b-2-3p-targeted HO-1 as a key player in the induction of ferroptosis by SiNPs, probably through regulating the intracellular iron metabolism to mediate iron overload and ensuing lipid peroxidation.

Conclusions

Our findings firstly underscored the fact that ferroptosis mediated by miR-125b-2-3p/HO-1 signaling was a contributor to SiNPs-induced myocardial injury, which could be of importance to elucidate the toxicity and provide new insights into the future safety applications of SiNPs-related nano products.

Graphical Abstract
A system for the study of roots 3D kinematics in hydroponic culture: a study on the oscillatory features of root tip
Authors: Simonetti Valentina , Ravazzolo Laura , Ruperti Benedetto , Quaggiotti Silvia , Castiello Umberto
Published: April 2024

Background

The root of a plant is a fundamental organ for the multisensory perception of the environment. Investigating root growth dynamics as a mean of their interaction with the environment is of key importance for improving knowledge in plant behaviour, plant biology and agriculture. To date, it is difficult to study roots movements from a dynamic perspective given that available technologies for root imaging focus mostly on static characterizations, lacking temporal and three-dimensional (3D) spatial information. This paper describes a new system based on time-lapse for the 3D reconstruction and analysis of roots growing in hydroponics.

Results

The system is based on infrared stereo-cameras acquiring time-lapse images of the roots for 3D reconstruction. The acquisition protocol guarantees the root growth in complete dark while the upper part of the plant grows in normal light conditions. The system extracts the 3D trajectory of the root tip and a set of descriptive features in both the temporal and frequency domains. The system has been used on Zea mays L. (B73) during the first week of growth and shows good inter-reliability between operators with an Intra Class Correlation Coefficient (ICC) > 0.9 for all features extracted. It also showed measurement accuracy with a median difference of < 1 mm between computed and manually measured root length.

Conclusions

The system and the protocol presented in this study enable accurate 3D analysis of primary root growth in hydroponics. It can serve as a valuable tool for analysing real-time root responses to environmental stimuli thus improving knowledge on the processes contributing to roots physiological and phenotypic plasticity.
Biomimetic design and clinical application of Ti-6Al-4V lattice hemipelvis prosthesis for pelvic reconstruction
Authors: Li Zhuangzhuang , Luo Yi , Lu Minxun , Wang Yitian , Gong Taojun , He Xuanhong , Hu Xin , Long Jingjunjiao , Zhou Yong , Min Li , Tu Chongqi
Published: April 2024

Objective

This study aims to biomimetic design a new 3D-printed lattice hemipelvis prosthesis and evaluate its clinical efficiency for pelvic reconstruction following tumor resection, focusing on feasibility, osseointegration, and patient outcomes.

Methods

From May 2020 to October 2021, twelve patients with pelvic tumors underwent tumor resection and subsequently received 3D-printed lattice hemipelvis prostheses for pelvic reconstruction. The prosthesis was strategically incorporated with lattice structures and solid to optimize mechanical performance and osseointegration. The pore size and porosity were analyzed. Patient outcomes were assessed through a combination of clinical and radiological evaluations.

Results

Multiple pore sizes were observed in irregular porous structures, with a wide distribution range (approximately 300–900 μm). The average follow-up of 34.7 months, ranging 26 from to 43 months. One patient with Ewing sarcoma died of pulmonary metastasis 33 months after surgery while others were alive at the last follow-up. Postoperative radiographs showed that the prosthesis’s position was consistent with the preoperative planning. T-SMART images showed that the host bone was in close and tight contact with the prosthesis with no gaps at the interface. The average MSTS score was 21 at the last follow-up, ranging from 18 to 24. There was no complication requiring revision surgery or removal of the 3D-printed hemipelvis prosthesis, such as infection, screw breakage, and prosthesis loosening.

Conclusion

The newly designed 3D-printed lattice hemipelvis prosthesis created multiple pore sizes with a wide distribution range and resulted in good osteointegration and favorable limb function.
Mediastinal Rosai-Dorfman Disease with KRAS mutation case report and literature review
Authors: Zhang Wenyu , Fang Linan , Wang Jing , Ma Xiaobo , Hu Xintong , Liu Wei
Published: April 2024

Background

Rosai-Dorfman Disease (RDD) is a rare self-limiting histiocytosis, more prevalent in children and young adults. It typically manifests as painless bilateral massive cervical lymphadenopathy but may also extend to extra-nodal sites, with intrathoracic RDD noted in 2% of cases. Distinguishing mediastinal RDD from thymoma on imaging poses challenges, underscoring the reliance on pathological features and immunohistochemical staining for diagnosis.

Case presentation

Patient, male, 33 years old, underwent lung a CT revealing an enlarged round soft tissue shadow in the anterior superior mediastinum, compared to a year ago. Surgical resection removed the entire mass, thymus, and part of the pericardium, confirming RDD on pathology. Genetic testing using second-generation testing technology identified a KRAS gene point mutation.

Conclusions

No established treatment protocol currently exists for this disease. However, as genetic mutation research progresses, a novel therapeutic avenue is emerging: targeted therapy integrated with surgical interventions.
Genetic insights into the ‘sandwich fusion’ subtype of Klippel–Feil syndrome: novel FGFR2 mutations identified by 21 cases of whole-exome sequencing
Authors: Xu Nanfang , Hung Kan-Lin , Gong Xiaoli , Fan Dongwei , Tian Yinglun , Yan Ming , Wei Yuan , Wang Shenglin
Published: April 2024

Background

Klippel–Feil syndrome (KFS) is a rare congenital disorder characterized by the fusion of two or more cervical vertebrae during early prenatal development. This fusion results from a failure of segmentation during the first trimester. Although six genes have previously been associated with KFS, they account for only a small proportion of cases. Among the distinct subtypes of KFS, “sandwich fusion” involving concurrent fusion of C0-1 and C2-3 vertebrae is particularly noteworthy due to its heightened risk for atlantoaxial dislocation. In this study, we aimed to investigate novel candidate mutations in patients with “sandwich fusion.”

Methods

We collected and analyzed clinical data from 21 patients diagnosed with “sandwich fusion.” Whole-exome sequencing (WES) was performed, followed by rigorous bioinformatics analyses. Our focus was on the six known KFS-related genes (GDF3, GDF6, MEOX1, PAX1, RIPPLY2, and MYO18). Suspicious mutations were subsequently validated through in vitro experiments.

Results

Our investigation revealed two novel exonic mutations in the FGFR2 gene, which had not previously been associated with KFS. Notably, the c.1750A > G variant in Exon 13 of FGFR2 was situated within the tyrosine kinase domain of the protein, in close proximity to several established post-translational modification sites. In vitro experiments demonstrated that this certain mutation significantly impacted the function of FGFR2. Furthermore, we identified four heterozygous candidate variants in two genes (PAX1 and MYO18B) in two patients, with three of these variants predicted to have potential clinical significance directly linked to KFS.

Conclusions

This study encompassed the largest cohort of patients with the unique “sandwich fusion” subtype of KFS and employed WES to explore candidate mutations associated with this condition. Our findings unveiled novel variants in PAX1, MYO18B, and FGFR2 as potential risk mutations specific to this subtype of KFS.
Identification of novel pathogenic variants of Calpain-3 gene in limb girdle muscular dystrophy R1
Authors: Banerjee Sukanya , Radotra Bishan Dass , Luthra-Guptasarma Manni , Goyal Manoj K
Published: April 2024

Background

Limb Girdle Muscular Dystrophy R1 (LGMDR1) is an autosomal recessive neuromuscular disease caused by mutations in the calpain-3 (CAPN3) gene. As clinical and pathological features may overlap with other types of LGMD, therefore definite molecular diagnosis is required to understand the progression of this debilitating disease. This study aims to identify novel variants of CAPN3 gene in LGMDR1 patients.

Results

Thirty-four patients with clinical and histopathological features suggestive of LGMD were studied. The muscle biopsy samples were evaluated using Enzyme histochemistry, Immunohistochemistry, followed by Western Blotting and Sanger sequencing. Out of 34 LGMD cases, 13 patients were diagnosed as LGMDR1 by immunoblot analysis, demonstrating reduced or absent calpain-3 protein as compared to controls. Variants of CAPN3 gene were also found and pathogenicity was predicted using in-silico prediction tools. The CAPN3 gene variants found in this study, included, two missense variants [CAPN3: c.1189T > C, CAPN3: c.2338G > C], one insertion-deletion [c.1688delinsTC], one splice site variant [c.2051-1G > T], and one nonsense variant [c.1939G > T; p.Glu647Ter].

Conclusions

We confirmed 6 patients as LGMDR1 (with CAPN3 variants) from our cohort and calpain-3 protein expression was significantly reduced by immunoblot analysis as compared to control. Besides the previously known variants, our study found two novel variants in CAPN3 gene by Sanger sequencing-based approach indicating that genetic variants in LGMDR1 patients may help to understand the etiology of the disease and future prognostication.
The endotoxin hypothesis of Alzheimer’s disease
Authors: Brown Guy C. , Heneka Michael T.
Published: April 2024

Lipopolysaccharide (LPS) constitutes much of the surface of Gram-negative bacteria, and if LPS enters the human body or brain can induce inflammation and act as an endotoxin. We outline the hypothesis here that LPS may contribute to the pathophysiology of Alzheimer’s disease (AD) via peripheral infections or gut dysfunction elevating LPS levels in blood and brain, which promotes: amyloid pathology, tau pathology and microglial activation, contributing to the neurodegeneration of AD. The evidence supporting this hypothesis includes: i) blood and brain levels of LPS are elevated in AD patients, ii) AD risk factors increase LPS levels or response, iii) LPS induces Aβ expression, aggregation, inflammation and neurotoxicity, iv) LPS induces TAU phosphorylation, aggregation and spreading, v) LPS induces microglial priming, activation and neurotoxicity, and vi) blood LPS induces loss of synapses, neurons and memory in AD mouse models, and cognitive dysfunction in humans. However, to test the hypothesis, it is necessary to test whether reducing blood LPS reduces AD risk or progression. If the LPS endotoxin hypothesis is correct, then treatments might include: reducing infections, changing gut microbiome, reducing leaky gut, decreasing blood LPS, or blocking LPS response.
A high-content screen of FDA approved drugs to enhance CAR T cell function: ingenol-3-angelate improves B7-H3-CAR T cell activity by upregulating B7-H3 on the target cell surface via PKCα activation
Authors: Lee Ha Won , O’Reilly Carla , Beckett Alex N. , Currier Duane G. , Chen Taosheng , DeRenzo Christopher
Published: April 2024

Background

CAR T cell therapy is a promising approach to improve outcomes and decrease toxicities for patients with cancer. While extraordinary success has been achieved using CAR T cells to treat patients with CD19-positive malignancies, multiple obstacles have so far limited the benefit of CAR T cell therapy for patients with solid tumors. Novel manufacturing and engineering approaches show great promise to enhance CAR T cell function against solid tumors. However, similar to single agent chemotherapy approaches, CAR T cell monotherapy may be unable to achieve high cure rates for patients with difficult to treat solid tumors. Thus, combinatorial drug plus CAR T cell approaches are likely required to achieve widespread clinical success.

Methods

We developed a novel, confocal microscopy based, high-content screen to evaluate 1114 FDA approved drugs for the potential to increase expression of the solid tumor antigen B7-H3 on the surface of osteosarcoma cells. Western blot, RT-qPCR, siRNA knockdown and flow cytometry assays were used to validate screening results and identify mechanisms of drug-induced B7-H3 upregulation. Cytokine and cytotoxicity assays were used to determine if drug pre-treatment enhanced B7-H3-CAR T cell effector function.

Results

Fifty-five drugs were identified to increase B7-H3 expression on the surface of LM7 osteosarcoma cells using a novel high-content, high-throughput screen. One drug, ingenol-3-angelate (I3A), increased B7-H3 expression by up to 100%, and was evaluated in downstream experiments. Validation assays confirmed I3A increased B7-H3 expression in a biphasic dose response and cell dependent fashion. Mechanistic studies demonstrated that I3A increased B7-H3 (CD276) mRNA, total protein, and cell surface expression via protein kinase C alpha activation. Functionally, I3A induced B7-H3 expression enhanced B7-H3-CAR T cell function in cytokine production and cytotoxicity assays.

Conclusions

This study demonstrates a novel high-content and high-throughput screen can identify drugs to enhance CAR T cell activity. This and other high-content technologies will pave the way to develop clinical trials implementing rational drug plus CAR T cell combinatorial therapies. Importantly, the technique could also be repurposed for an array of basic and translational research applications where drugs are needed to modulate cell surface protein expression.
“NeuroVanguard”: a contemporary strategy in neuromonitoring for severe adult brain injury patients
Authors: Rodriguez Edith Elianna , Zaccarelli Mario , Sterchele Elda Diletta , Taccone Fabio Silvio
Published: April 2024

Severe acute brain injuries, stemming from trauma, ischemia or hemorrhage, remain a significant global healthcare concern due to their association with high morbidity and mortality rates. Accurate assessment of secondary brain injuries severity is pivotal for tailor adequate therapies in such patients. Together with neurological examination and brain imaging, monitoring of systemic secondary brain injuries is relatively straightforward and should be implemented in all patients, according to local resources. Cerebral secondary injuries involve factors like brain compliance loss, tissue hypoxia, seizures, metabolic disturbances and neuroinflammation. In this viewpoint, we have considered the combination of specific noninvasive and invasive monitoring tools to better understand the mechanisms behind the occurrence of these events and enhance treatment customization, such as intracranial pressure monitoring, brain oxygenation assessment and metabolic monitoring. These tools enable precise intervention, contributing to improved care quality for severe brain injury patients. The future entails more sophisticated technologies, necessitating knowledge, interdisciplinary collaboration and resource allocation, with a focus on patient-centered care and rigorous validation through clinical trials.
A single-center, open-labeled, randomized, 6-month, parallel-group study to assess the safety and efficacy of allogeneic cultured keratinocyte sheet transplantation for deep second-degree burn wounds: rationale and design of phase I/II clinical trial
Authors: Farzanbakhsh Shayan , Shahrbaf Mohammad Amin , Madani Hoda , Dahmardei Mostafa , Sadri Bahareh , Vosough Massoud
Published: April 2024

Background

Burn-related injuries are a major global health issue, causing 180,000 deaths per year. Early debridement of necrotic tissue in association with a split-thickness skin graft is usually administered for some of the 2nd- and 3rd-degree injuries. However, this approach can be complicated by factors such as a lack of proper donor sites. Artificial skin substitutes have attracted much attention for burn-related injuries. Keratinocyte sheets are one of the skin substitutes that their safety and efficacy have been reported by previous studies.

Methods

Two consecutive clinical trials were designed, one of them is phase I, a non-randomized, open-label trial with 5 patients, and phase II is a randomized and open-label trial with 35 patients. A total number of 40 patients diagnosed with 2nd-degree burn injury will receive allogenic keratinocyte sheet transplantation. The safety and efficacy of allogeneic skin graft with autograft skin transplantation and conventional treatments, including Vaseline dressing and topical antibiotic, will be compared in different wounds of a single patient in phase II. After the transplantation, patients will be followed up on days 3, 7, 10, 14, 21, and 28. In the 3rd and 6th months after the transplantation scar, a wound closure assessment will be conducted based on the Vancouver Scar Scale and the Patient and Observer Scar Assessment Scale.

Discussion

This study will explain the design and rationale of a cellular-based skin substitute for the first time in Iran. In addition, this work proposes this product being registered as an off-the-shelf product for burn wound management in the country.

Trial registration

Iranian Registry of Clinical Trials (IRCT) IRCT20080728001031N31, 2022-04-23 for phase I and IRCT20080728001031N36, 2024-03-15 for phase II.
IL-33-ST2 signaling in fibro-adipogenic progenitors alleviates immobilization-induced muscle atrophy in mice
Authors: Takahashi Yoshiyuki , Yoda Masaki , Tsuji Osahiko , Horiuchi Keisuke , Watanabe Kota , Nakamura Masaya
Published: April 2024

Background

The regenerative and adaptive capacity of skeletal muscles reduces with age, leading to severe disability and frailty in the elderly. Therefore, development of effective therapeutic interventions for muscle wasting is important both medically and socioeconomically. In the present study, we aimed to elucidate the potential contribution of fibro-adipogenic progenitors (FAPs), which are mesenchymal stem cells in skeletal muscles, to immobilization-induced muscle atrophy.

Methods

Young (2–3 months), adult (12–14 months), and aged (20–22 months) mice were used for analysis. Muscle atrophy was induced by immobilizing the hind limbs with a steel wire. FAPs were isolated from the hind limbs on days 0, 3, and 14 after immobilization for transcriptome analysis. The expression of ST2 and IL-33 in FAPs was evaluated by flow cytometry and immunostaining, respectively. To examine the role of IL-33-ST2 signaling in vivo, we intraperitoneally administered recombinant IL-33 or soluble ST2 (sST2) twice a week throughout the 2-week immobilization period. After 2-week immobilization, the tibialis anterior muscles were harvested and the cross-sectional area of muscle fibers was evaluated.

Results

The number of FAPs increased with the progression of muscle atrophy after immobilization in all age-groups. Transcriptome analysis of FAPs collected before and after immobilization revealed that Il33 and Il1rl1 transcripts, which encode the IL-33 receptor ST2, were transiently induced in young mice and, to a lesser extent, in aged mice. The number of FAPs positive for ST2 increased after immobilization in young mice. The number of ST2-positive FAPs also increased after immobilization in aged mice, but the difference from the baseline was not statistically significant. Immunostaining for IL-33 in the muscle sections revealed a significant increase in the number of FAPs expressing IL-33 after immobilization. Administration of recombinant IL-33 suppressed immobilization-induced muscle atrophy in aged mice but not in young mice.

Conclusions

Our data reveal a previously unknown protective role of IL-33-ST2 signaling against immobilization-induced muscle atrophy in FAPs and suggest that IL-33-ST2 signaling is a potential new therapeutic target for alleviating disuse muscle atrophy, particularly in older adults.
Complement C1q-mediated microglial synaptic elimination by enhancing desialylation underlies sevoflurane-induced developmental neurotoxicity
Authors: Wang Gang , Liu Hua-yue , Meng Xiao-wen , Chen Ying , Zhao Wei-ming , Li Wen-ting , Xu Han-bing , Peng Ke , Ji Fu-hai
Published: April 2024

Background

Repeated neonatal sevoflurane exposures led to neurocognitive disorders in young mice. We aimed to assess the role of microglia and complement C1q in sevoflurane-induced neurotoxicity and explore the underlying mechanisms.

Methods

Neonatal mice were treated with sevoflurane on postnatal days 6, 8, and 10, and the Morris water maze was performed to assess cognitive functions. For mechanistic explorations, mice were treated with minocycline, C1q-antibody ANX005, and sialidase-inhibitor N-acetyl-2,3-dehydro-2-deoxyneuraminic acid (NADNA) before sevoflurane exposures. Western blotting, RT-qPCR, Golgi staining, 3D reconstruction and engulfment analysis, immunofluorescence, and microglial morphology analysis were performed. In vitro experiments were conducted in microglial cell line BV2 cells.

Results

Repeated neonatal sevoflurane exposures resulted in deficiencies in learning and cognition of young mice, accompanied by microglial activation and synapse loss. Sevoflurane enhanced microglia-mediated synapse elimination through C1q binding to synapses. Inhibition of microglial activation and phagocytosis with minocycline significantly reduced the loss of synapses. We further revealed the involvement of neuronal sialic acids in this process. The enhanced activity of sialidase by sevoflurane led to the loss of sialic acids, which facilitated C1q binding to synapses. Inhibition of C1q with ANX005 or inhibition of sialidase with NADNA significantly rescued microglia-mediated synapse loss and improved neurocognitive function. Sevoflurane enhanced the engulfment of BV2 cells, which was reversed by ANX005.

Conclusions

Our findings demonstrated that C1q-mediated microglial synaptic elimination by enhancing desialylation contributed to sevoflurane-induced developmental neurotoxicity. Inhibition of C1q or sialidase may be a potential therapeutic strategy for this neurotoxicity.
Rapid recovery of desert shrubs is attributed to imbalance of water supply to demand after disturbance in Northwest China
Authors: Zhou Hai , Feng Xiangyan , He Zhibin , Yang Qiyue , Ren Heng , Zhao Wenzhi
Published: April 2024

Background

Disturbance is a key driver of changes in terrestrial ecosystems, especially in water limited regions. In the desert ecosystem of Northwest China, shrubs are often foraged by camels and this causes the shrubs lost their aboveground tissues. Although the aboveground tissue could recover after the disturbance, the mechanisms of resprouts’ recovery after the disturbance are still poorly understood. We investigated the morphological and physiological characteristics and root water uptake sources of resprouts and non-disturbed seedlings of four desert shrub species that were often foraged by camels.

Results

The results showed that the resprouts grew rapidly after the removal of aboveground tissue. The height and aboveground biomass of resprouts reached the level of 5-year-old seedlings following a growing season. Compared to un-disturbed seedlings, higher leaf water potential, leaf hydraulic conductance and photosynthesis rate were observed in resprouts, indicating that resprouts grew rapidly after disturbance. Furthermore, we found that the root water uptake source did not change after disturbance. This indicated that the roots of resprouts kept intact water uptake capacity after disturbance, which could keep supplying sufficient water to the aboveground tissue. Due to the removal of aboveground tissue of the resprouts, the water demand of aboveground tissue decreased, which induced an imbalance of water supply from roots to demand of aboveground tissues of resprouts. This explained the improved physiological performance and rapid recovery of resprouts after disturbance.

Conclusions

The imbalance of water supply to demand contributes to the rapid recovery of desert shrubs after disturbance. The findings provide insight into the eco-physiological mechanism for the maintenance of desert ecosystems that are facing disturbance.
Piperine reduces hair oiliness by inhibiting adipogenesis of hair stem cells
Authors: Im Minyoung , Kim Nackhyoung , Park Ui-Hyun , Heo Hyeon Ho , Um Soo-Jong
Published: April 2024

Piperine, an alkaloid compound in black pepper (Piper nigrum), has beneficial bioactivities. Specifically, piperine inhibits adipogenesis in 3T3-L1 cells by suppressing the transcriptional activity of PPARγ. Control of hair oiliness, which is related to adipogenic regulation, is important to prevent hair loss. Excessive sebum from the sebaceous gland (SG) can cause acne, folliculitis, or irritated skin by clogging pores. To investigate the in vivo function of piperine in SG, we used mice fed a high-fat diet (HFD). The HFD increased the size and Oil Red O (ORO) staining intensity of SG, which were significantly reduced by piperine. The HFD also upregulated the expression of sebocyte-associated genes, including PPARγ target genes, an effect reversed by piperine. In CD34/CD49f double-positive hair follicle bulge stem cells isolated from mouse vibrissae, piperine inhibited cellular adipogenesis, likely via transcriptional repression of related genes. Furthermore, piperine reduced the thickness of subcutaneous fat. In human dermal papilla cells, piperine inhibited cellular adipogenesis, as shown by the reduction in ORO staining and the downregulation of PPARγ target genes. In conclusion, piperine can be used to reduce hair greasiness by suppressing adipogenesis in hair stem cells.
Classification of multiple text reading test formats by analyzing English language proficiency tests
Authors: Mikami Yosuke
Published: April 2024

This study proposes a classification method for multiple text reading test formats in English language proficiency tests. A preliminary study involving 11 proficiency tests revealed two tests that fit the scope of the main study. Results show that multiple text reading test formats use complementary texts rather than conflicting texts. As for questions in a set of test forms in multiple text reading test formats, cognitive processing on integrating the contents of texts was different in proficiency tests. Moreover, the type of connection formation required by the questions differed among the proficiency tests. Implications for pedagogy are presented.
Empowering students through active learning in educational big data analytics
Authors: Tsai Yun-Cheng
Published: April 2024

Purpose

This paper explores how Educational Big Data Analytics can enhance student learning. It investigates the role of active learning in improving students’ data analysis skills and critical thinking. By actively engaging students in data analysis assessments, the aim is to equip them with the skills to navigate the data-rich educational landscape.

Methods

The study uses a teaching strategy that combines structured and unstructured data analysis using Python tools and ChatGPT APIs. It presents five assignments, each highlighting data analysis skills and encouraging critical thinking.

Results

The paper offers insights into how the teaching strategy effectively enhances students’ data analysis and critical thinking skills. It investigates the specific impact of active learning on students’ engagement with educational data. The study reveals that all students can complete a comprehensive project, integrating the skills they have learned in the five assignments related to educational big data while incorporating the educational implications from their respective disciplines.

Conclusion

The key lies in instructors being able to design individual assignments that link practical experiences, enabling each teaching session’s effectiveness to accumulate in students’ personal experiences and practical skills, ultimately empowering them with the abilities necessary to work effectively with Educational Big Data Analytics. The findings of this study make a valuable contribution to the ongoing conversation about enhancing the educational experience for students in this data-rich era.
Optimizing PSMA scintigraphy for resource limited settings – a retrospective comparative study
Authors: Kolade Olumayowa U. , Brink Anita , Ayeni Akinwale O. , More Stuart , Holness Jennifer
Published: April 2024

Background

PSMA PET/CT is the most sensitive molecular imaging modality for prostate cancer (PCa), yet much of the developing world has little or no access to PET/CT. [99mTc]Tc-PSMA scintigraphy (PS) is a cheaper and more accessible gamma camera-based alternative. However, many resource-constrained departments have only a single camera without tomographic or hybrid imaging functionality, and camera time is frequently in high demand. Simplifying imaging protocols by limiting the field of view (FOV) and omitting SPECT/CT or even SPECT may provide a partial solution. The aim was thus to determine the adequacy of PS planar-only and/or SPECT-only imaging protocols with a limited FOV.

Methods

The scans of 95 patients with histologically proven PCa who underwent PS with full-body planar and multi-FOV SPECT/CT were reviewed. The detection rates for uptake in the prostate gland/bed and in metastases were compared on planar, SPECT, and SPECT/CT. The agreement between modalities was calculated for the detection of metastases and for staging. The impact of imaging a limited FOV was determined.

Results

Pathological prostatic uptake was seen in all cases on SPECT/CT (excluding two post-prostatectomy patients), 90.3% of cases on SPECT, and 15.1% on planar images (p < 0.001). Eleven (11.7%) patients had seminal vesicle involvement on SPECT/CT, which was undetectable/indistinguishable on planar images and SPECT. The agreement between modalities was moderate to good (κ = 0.41 to 0.61) for the detection of nodal metastases, with detection rates that did not differ significantly (SPECT/CT = 11.6%, SPECT = 8.4%, planar = 5.3%). Detection rates for bone metastases were 14.7% (SPECT/CT) and 11.6% (SPECT and planar). Agreement between modalities for the detection of bone metastases was good (κ = 0.73 to 0.77). Three (3.1%) patients had visceral metastases on SPECT/CT, two of which were detected on SPECT and planar. There was good agreement between modalities for the TNM staging of patients (κ = 0.70 to 0.88). No metastatic lesions were missed on the limited FOV images.

Conclusion

When PS scintigraphy is performed, SPECT/CT is recommended. However, the lack of SPECT/CT capabilities should not preclude the use of PS in the presence of limited resources, as both planar and SPECT imaging are adequate and will correctly stage most PCa patients. Furthermore, time-based optimisations are achievable by limiting the FOV to exclude the distal lower limbs.
Non-diffractive, toric, extended depth-of-focus intraocular lenses in eyes with low corneal astigmatism
Authors: Pastor-Pascual Francisco , Orts-Vila Paz , Tañá-Sanz Pedro , Tañá-Sanz Santiago , Ruiz-Mesa Ramón , Tañá-Rivero Pedro
Published: April 2024

Background

To assess clinical outcomes after implanting toric, extended-depth-of-focus intraocular lenses (IOLs) to correct low corneal astigmatism in eyes with cataracts.

Methods

47 eyes were implanted with the AcrySof IQ Vivity Toric DFT215 IOL. Main outcome measures were refractive error, monocular uncorrected and corrected distance (UDVA/CDVA), uncorrected and distance-corrected intermediate (UIVA/DCIVA), and uncorrected near and distance-corrected near (UNVA/DCNVA) visual acuities, monocular defocus curve, rotational stability, and IOLSAT and QUVID questionnaires. Patients were assessed at 3 months postsurgery.

Results

All eyes had a postoperative spherical equivalent (SE) within ± 0.50 D and 97.87% (n = 46) had a refractive cylinder ≤ 0.50 D. The mean SE and refractive cylinder were − 0.10 ± 0.17 D and − 0.16 ± 0.24 D, respectively. The CDVA was ≥ 20/25 and ≥ 20/32 in 95.74% (n = 45) and 97.87% (n = 46) of eyes, respectively. The DCIVA was ≥ 20/32 in 85.11% (n = 40) of eyes and the DCNVA was ≥ 20/40 in 74.47% (n = 35). The mean values of CDVA, DCIVA, and DCNVA were − 0.02 ± 0.08, 0.14 ± 0.09, and 0.23 ± 0.12 logMAR, respectively. The defocus curve revealed good visual acuity at far and intermediate distances with a depth-of-focus of about 1.75 D. IOL rotation was 0.74 ± 1.13 degrees and all eyes had a rotation of less than 5 degrees. Patients reported either good or very good postoperative vision without eyeglasses under bright-light-conditions at distance (87.80%, 36/41) and intermediate distance (92.68%, 38/41). Between about 63.83%–72.34% (30–34) of patients reported no starburst, halos, or glare, or if experienced, were not bothersome.

Conclusions

The Vivity toric IOL implanted in eyes with low-astigmatism provides accurate refractive outcomes, good visual acuity at different distances and excellent rotational stability.

Trial Registration The study was registered with the German Clinical Trials Register (DRKS00030579)