bluuebunny/crossref_metadata_2025_split huggingface dataset:
Targeted control of supporting pathways in paclitaxel biosynthesis with CRISPR-guided methylation
Authors: Brzycki Newton Cassandra , Young Eric M. , Roberts Susan C. 
Published: October 2023

Introduction: Plant cell culture biomanufacturing is rapidly becoming an effective strategy for production of high-value plant natural products, such as therapeutic proteins and small molecules, vaccine adjuvants, and nutraceuticals. Many of these plant natural products are synthesized from diverse molecular building blocks sourced from different metabolic pathways. Even so, engineering approaches for increasing plant natural product biosynthesis have typically focused on the core biosynthetic pathway rather than the supporting pathways.Methods: Here, we use both CRISPR-guided DNA methylation and chemical inhibitors to control flux through the phenylpropanoid pathway in Taxus chinensis, which contributes a phenylalanine derivative to the biosynthesis of paclitaxel (Taxol), a potent anticancer drug. To inhibit PAL, the first committed step in phenylpropanoid biosynthesis, we knocked down expression of PAL in Taxus chinensis plant cell cultures using a CRISPR-guided plant DNA methyltransferase (NtDRM). For chemical inhibition of downstream steps in the pathway, we treated Taxus chinensis plant cell cultures with piperonylic acid and caffeic acid, which inhibit the second and third committed steps in phenylpropanoid biosynthesis: cinnamate 4-hydroxylase (C4H) and 4-coumaroyl-CoA ligase (4CL), respectively.Results: Knockdown of PAL through CRISPR-guided DNA methylation resulted in a profound 25-fold increase in paclitaxel accumulation. Further, through the synergistic action of both chemical inhibitors and precursor feeding of exogenous phenylalanine, we achieve a 3.5-fold increase in paclitaxel biosynthesis and a similar reduction in production of total flavonoids and phenolics. We also observed perturbations to both activity and expression of PAL, illustrating the complex transcriptional co-regulation of these first three pathway steps.Discussion: These results highlight the importance of controlling the metabolic flux of supporting pathways in natural product biosynthesis and pioneers CRISPR-guided methylation as an effective method for metabolic engineering in plant cell cultures. Ultimately, this work demonstrates a powerful method for rewiring plant cell culture systems into next-generation chassis for production of societally valuable compounds.
Elevated CO2 does not improve seedling performance in a naturally regenerated oak woodland exposed to biotic stressors
Authors: Mayoral Carolina , Ioni Susanna , Luna Estrella , Crowley Liam M. , Hayward Scott A. L. , Sadler Jon P. , MacKenzie A. Robert 
Published: October 2023

IntroductionInsect herbivores and biotrophic pathogens are major stressors influencing natural regeneration in woodlands. Information on the effect of elevated CO2 (eCO2) on plant-insect-pathogen interactions under natural conditions is lacking.MethodsWe studied the effects of eCO2 on leaf-out phenology, as well as on levels of insect herbivory and powdery mildew (PM), i.e., reduction of leaf photosynthetic material. We then assessed the combined impacts of these biotic stressors and eCO2 on seedling photosynthesis and growth. A total of 92 naturally recruited and 114 potted seedlings of 5 temperate tree species (Quercus robur, Acer pseudoplatanus, Corylus avellana, Crataegus monogyna, and Ilex aquifolium) within a mature oak woodland were studied.ResultsWe found that eCO2 advanced leaf-out phenology and was a significant explanatory variable for growth and physiological performance in potted seedlings. Potted oak seedlings experienced 11-fold higher tissue loss from insect herbivory than natural seedlings. The earliest leaf-flushing species, hawthorn, and the evergreen holly were resistant to insect attack and were not affected by PM. Oak was defoliated most but showed the highest regeneration capacity. Hazel was more resistant to PM infection than oak and sycamore. Despite being highly infected by PM, sycamore was less affected than oak. The more vigorous sycamore and oak seedlings suffered more severe PM disease.ConclusionNo evidence emerged that eCO2 enhances natural regeneration under biotic stress for any of the species studied.
Omicron subvariants illustrate reduced respiratory tissue penetration, cell damage and inflammatory responses in human airway epithelia
Authors: Zaderer Viktoria , Abd El Halim Hussam , Wyremblewsky Anna-Lena , Lupoli Gaia , Dächert Christopher , Muenchhoff Maximilian , Graf Alexander , Blum Helmut , Lass-Flörl Cornelia , Keppler Oliver T. , Huber Lukas A. , Posch Wilfried , Wilflingseder Doris 
Published: October 2023

IntroductionTo explore whether the reported lower pathogenicity in infected individuals of variant of concern (VoC) Omicron and its current subvariants compared to VoC Delta may be related to fundamental differences in the initial virus-tissue interaction, we assessed their ability to penetrate, replicate and cause damage in a human 3D respiratory model.MethodsFor this, we used TEER measurements, real-time PCR, LDH, cytokine and complex confocal imaging analyses.Results and discussionWe observed that Delta readily penetrated deep into the respiratory epithelium and this was associated with major tissue destruction, high LDH activity, high viral loads and pronounced innate immune activation as observed by intrinsic C3 activation and IL-6 release at infection sites. In contrast, Omicron subvariants BA.5, BQ.1.1 and BF7 remained superficially in the mucosal layer resulting merely in outward-directed destruction of cells, maintenance of epithelial integrity, minimal LDH activity and low basolateral release of virus at infection sites, as well as significantly smaller areas of complement activation and lower IL-6 secretion. Interestingly, also within Omicron subvariants differences were observed with newer Omicron subvariants BQ.1.1 and BF.7 illustrating significantly reduced viral loads, IL-6 release and LDH activity compared to BA.5. Our data indicate that earliest interaction events after SARS-CoV-2 transmission may have a role in shaping disease severity.
Single cell transcriptomics reveals cell type specific features of developmentally regulated responses to lipopolysaccharide between birth and 5 years
Authors: Read James F. , Serralha Michael , Armitage Jesse D. , Iqbal Muhammad Munir , Cruickshank Mark N. , Saxena Alka , Strickland Deborah H. , Waithman Jason , Holt Patrick G. , Bosco Anthony 
Published: October 2023

BackgroundHuman perinatal life is characterized by a period of extraordinary change during which newborns encounter abundant environmental stimuli and exposure to potential pathogens. To meet such challenges, the neonatal immune system is equipped with unique functional characteristics that adapt to changing conditions as development progresses across the early years of life, but the molecular characteristics of such adaptations remain poorly understood. The application of single cell genomics to birth cohorts provides an opportunity to investigate changes in gene expression programs elicited downstream of innate immune activation across early life at unprecedented resolution.MethodsIn this study, we performed single cell RNA-sequencing of mononuclear cells collected from matched birth cord blood and 5-year peripheral blood samples following stimulation (18hrs) with two well-characterized innate stimuli; lipopolysaccharide (LPS) and Polyinosinic:polycytidylic acid (Poly(I:C)).ResultsWe found that the transcriptional response to LPS was constrained at birth and predominantly partitioned into classical proinflammatory gene upregulation primarily by monocytes and Interferon (IFN)-signaling gene upregulation by lymphocytes. Moreover, these responses featured substantial cell-to-cell communication which appeared markedly strengthened between birth and 5 years. In contrast, stimulation with Poly(I:C) induced a robust IFN-signalling response across all cell types identified at birth and 5 years. Analysis of gene regulatory networks revealed IRF1 and STAT1 were key drivers of the LPS-induced IFN-signaling response in lymphocytes with a potential developmental role for IRF7 regulation.ConclusionAdditionally, we observed distinct activation trajectory endpoints for monocytes derived from LPS-treated cord and 5-year blood, which was not apparent among Poly(I:C)-induced monocytes. Taken together, our findings provide new insight into the gene regulatory landscape of immune cell function between birth and 5 years and point to regulatory mechanisms relevant to future investigation of infection susceptibility in early life.
A human pilot study on positive electrostatic charge effects in solid tumors of the late-stage metastatic patients
Authors: Zandi Ashkan , Shojaeian Fatemeh , Abbasvandi Fereshteh , Faranoush Mohammad , Anbiaee Robab , Hoseinpour Parisa , Gilani Ali , Saghafi Mohammad , Zandi Afsoon , Hoseinyazdi Meisam , Davari Zahra , Miraghaie Seyyed Hossein , Tayebi Mahtab , Taheri Morteza Sanei , Ardestani S. Mehdi Samimi , Sheikhi Mobarakeh Zahra , Nikshoar Mohammad Reza , Enjavi Mohammad Hossein , Kordehlachin Yasin , Mousavi-kiasary S. M. Sadegh , Mamdouh Amir , Akbari Mohammad Esmaeil , Yunesian Masud , Abdolahad Mohammad 
Published: October 2023

BackgroundCorrelative interactions between electrical charges and cancer cells involve important unknown factors in cancer diagnosis and treatment. We previously reported the intrinsic suppressive effects of pure positive electrostatic charges (PEC) on the proliferation and metabolism of invasive cancer cells without any effect on normal cells in cell lines and animal models. The proposed mechanism was the suppression of pro-caspases 3 and 9 with an increase in Bax/Bcl2 ratio in exposed malignant cells and perturbation induced in the KRAS pathway of malignant cells by electrostatic charges due to the phosphate molecule electrostatic charge as the trigger of the pathway. This study aimed to examine PECs as a complementary treatment for patients with different types of solid metastatic tumors, who showed resistance to chemotherapy and radiotherapy.MethodsIn this study, solid metastatic tumors of the end-stage patients (n = 41) with various types of cancers were locally exposed to PEC for at least one course of 12 days. The patient’s signs and symptoms, the changes in their tumor size, and serum markers were followed up from 30 days before positive electrostatic charge treating (PECT) until 6 months after the study.ResultsEntirely, 36 patients completed the related follow-ups. Significant reduction in tumor sizes and cancer-associated enzymes as well as improvement in cancer-related signs and symptoms and patients’ lifestyles, without any side effects on other tissues or metabolisms of the body, were observed in more than 80% of the candidates.ConclusionPECT induced significant cancer remission in combination with other therapies. Therefore, this non-ionizing radiation would be a beneficial complementary therapy, with no observable side effects of ionizing radiotherapy, such as post-radiation inflammation.
Midgut microbiota affects the intestinal barrier by producing short-chain fatty acids in Apostichopus japonicus
Authors: Song Mingshan , Zhang Zhen , Li Yanan , Xiang Yangxi , Li Chenghua 
Published: October 2023

IntroductionThe intestinal microbiota participates in host physiology and pathology through metabolites, in which short-chain fatty acids (SCFAs) are considered principal products and have extensive influence on intestine homeostasis. It has been reported that skin ulceration syndrome (SUS), the disease of Apostichopus japonicus caused by Vibrio splendidus, is associated with the alteration of the intestinal microbiota composition.MethodTo investigate whether the intestinal microbiota affects A. japonicus health via SCFAs, in this study, we focus on the SCFA profiling and intestinal barrier function in A. japonicus treated with V. splendidus.Results and discussionWe found that V. splendidus could destroy the mid-intestine integrity and downregulate the expression of tight junction proteins ZO-1 and occludin in A. japonicus, which further dramatically decreased microorganism abundance and altered SCFAs contents. Specifically, acetic acid is associated with the largest number of microorganisms and has a significant correlation with occludin and ZO-1 among the seven SCFAs. Furthermore, our findings showed that acetic acid could maintain the intestinal barrier function by increasing the expression of tight junction proteins and rearranging the tight junction structure by regulating F-actin in mid-intestine epithelial cells. Thus, our results provide insights into the effects of the gut microbiome and SCFAs on intestine barrier homeostasis and provide essential knowledge for intervening in SUS by targeting metabolites or the gut microbiota.
10-hydroxy-2-decenoic acid alleviates lipopolysaccharide-induced intestinal mucosal injury through anti-inflammatory, antioxidant, and gut microbiota modulation activities in chickens
Authors: Han Lianquan , Zhang Maolu , Li Fuwei , Su Jing , Wang Ruiming , Li Guiming , Yang Xiaohui 
Published: October 2023

IntroductionThis study aimed to investigated the effects of 10-hydroxy-2-decenoic acid (10-HDA) on the growth performance, intestinal barrier, inflammatory response, oxidative stress, and gut microbiota of chickens challenged with lipopolysaccharide (LPS).MethodsA total of 240 one-day-old chickens were randomly assigned to five treatment groups: (1) control group (basal diet + saline); (2) LPS group (basal diet + LPS); (3) Chlortetracycline (CTC) group (basal diet containing 75 mg/kg CTC + LPS); (4) 0.1% 10-HDA group (basal diet containing 1 g/kg 10-HDA + LPS); and (5) 0.5% 10-HDA group (basal diet containing 5 g/kg 10-HDA + LPS). All chickens were injected intraperitoneally with 0.5 mg/kg body weight of either LPS or saline at 17, 19, and 21 days of age.ResultsThe results showed that dietary 10-HDA supplementation attenuated the loss in growth performance caused by the LPS challenge (p &lt; 0.05). 10-HDA effectively alleviated LPS-induced intestinal mucosal injury, as evidenced by reduced bleeding, decreased serum diamine oxidase levels (p &lt; 0.05), and increased villus/crypt ratios of the jejunum and ileum (p &lt; 0.05). Dietary treatment with 0.1% 10-HDA reduced the concentrations of inflammatory cytokines (TNF-α, IL-1β, IL-6; p &lt; 0.05), and increased immunoglobulin (IgA, IgG) and antioxidant enzyme levels (CAT, GSH-px, T-SOD) in the serum of LPS-challenged chickens (p &lt; 0.05). These effects were similar to those observed in the CTC group. Moreover, 0.1% 10-HDA treatment reversed the LPS-induced variations in the mRNA expression of genes related to inflammation, antioxidant capacity, and intestinal tight junctions (p &lt; 0.05). 16S rRNA analysis revealed that 10-HDA supplementation increased the relative abundance of Faecalibacterium and Clostridia_UCG-014 (p &lt; 0.05). Additionally, it decreased the abundance of Clostridia_vadinBB60_group, Eubacterium_nodatum_group, and UC5-1-2E3 (p &lt; 0.05). These changes were correlated with reduced inflammation and improved antioxidant capacity in the LPS-challenged chickens.ConclusionCollectively, dietary 10-HDA supplementation alleviated LPS-induced intestinal mucosal injury and the loss of growth performance through anti-inflammatory, antioxidant, and gut microbiota modulation activities in chickens. Moreover, 0.1% 10-HDA supplementation had comparable or even better protection for LPS-challenged chickens than supplementation with antibiotics or 0.5% 10-HDA. 10-HDA has the potential to be used as an alternative to antibiotics in protecting the intestinal health and improving the performance of poultry.
Comorbidity and outcomes among hospitalized patients with stroke: a nationwide inpatient analysis
Authors: Chen Wei , Li Dong 
Published: October 2023

ObjectiveWe aimed to characterize healthcare utilization and comorbidity outcomes among hospitalized elderly stroke patients using a nationally representative dataset in the United States.MethodsUsing the 2019 National Inpatient Sample, patients aged 65 years or older with and without comorbidities who were hospitalized for acute stroke were identified using the International Classification of Diseases, Tenth Revision, Clinical Modification codes. Patient comorbidities were identified with the use of the Elixhauser Comorbidity Index. The prevalence of comorbidities, in-hospital mortality, length of stay, and total hospital costs were analyzed for these patients.ResultsWithin 451,945 patients (mean age 78 years, 54.1% women, 73.7% white), we observed that more than 90% of patients had a minimum of two comorbidities. The median number of comorbidities was 4.0 (IQR 3.0–6.0). There was significant variation in the prevalence rate of comorbidities. The five most common comorbidities were uncomplicated hypertension (55.4%), paralysis (40.1%), congestive heart failure (39.8%), various neurological illnesses (38.3%), and complex hypertension (32.5%). After adjusting for patient- and hospital-level characteristics, a statistically significant association was observed between comorbidities and various adverse outcomes. Specifically, comorbidities were found to be significantly associated with an increased risk of inpatient mortality (odds ratio: 1.09; 95% CI: 1.08–1.11; P &lt; 0.001), a longer duration of hospitalization (0.68 days; 95% CI: 0.66–0.71; P &lt; 0.001), and higher total cost ($1,874.9; 95% CI: 1,774.6–1,975.2; P &lt; 0.001).ConclusionThis national data suggests that comorbidity is common among hospitalized older stroke patients and substantially increases the healthcare burden and inpatient mortality in the United States. These findings underscore the integration of comorbidity management into the care of older stroke patients.
Alcoholic drink produced by pea is a risk factor for incident knee surgery in patients with knee osteoarthritis
Authors: Huang Xiaopeng , Zhang Jinshan , Zheng Yongqiang , Liu Xiaofeng , Xu Yongquan , Fang Yangzhen , Lin Zhenyu , Lin Liang , Zhang Hongpeng , Wang Zefeng 
Published: October 2023

ObjectiveThe objective of this study is to investigate whether alcohol exposure and specific alcoholic drinks are independent risk factors for incident knee surgery in knee osteoarthritis (KOA) patients.MethodsWe identified all patients who were clinically diagnosed as KOA between January 2010 and January 2018 in our outpatient department. Demographic, clinical, and radiographic data were collected from the database of our hospital. Next, we analyzed the association between alcohol consumption and incident knee surgery.ResultsA total of 4,341 KOA patients completed the current study and were included in the final analysis. Incident knee surgery for the purpose of treating osteoarthritis was observed in 242 patients. Incident knee surgery was significantly associated with age (OR [95%CI], 1.023 [1.009–1.039], P = 0.002), BMI (OR [95%CI], 1.086 [1.049–1.123], P &lt; 0.001), baseline K-L grade 3 (OR [95%CI], 1.960 [1.331–2.886], P = 0.001), baseline K-L grade 4 (OR [95%CI], 1.966 [1.230–3.143], P = 0.005), 7.1–14 drinks per week (OR [95%CI], 2.013 [1.282–3.159], P = 0.002), &gt;14 standard drinks per week (OR [95%CI], 2.556 [1.504–4.344], P = 0.001), and the most common alcoholic drink produced by pea (OR [95%CI], 3.133 [1.715–5.723], P &lt; 0.001).ConclusionKOA patients who consumed more than seven standard drinks per week were at substantial risk of incident knee surgery. In addition, alcoholic drink produced by pea is also an independent risk factor.
Application of endoport-assisted neuroendoscopic techniques in lateral ventricular tumor surgery
Authors: Yan Chaolong , Mao Jiannan , Yao Chenbei , Liu Yang , Jin Wei , Yan Huiying 
Published: October 2023

ObjectiveThe objective of this study was to investigate the clinical experience and therapeutic efficiency of Endoport-assisted neuroendoscopic surgery for resection of lateral ventricular tumors. The key points and application value of this surgical technique were additionally discussed.MethodsA retrospective analysis was conducted on the clinical and follow-up data of 16 patients who underwent endoport-assisted neuroendoscopic surgery for lateral ventricular tumors at the Department of Neurosurgery, Nanjing Drum Tower Hospital, the Affiliated Hospital of Nanjing University Medical School, between January 2018 and September 2020. The surgical procedures, complications and outcomes were analyzed.ResultsThe study included a total of 16 patients (5 males and 11 females) with lateral ventricular tumors, with a mean age of 43.2 years (18-70 years old). The tumors were distributed as follows: 5 cases involved the body of the lateral ventricle, 3 involved the frontal horn and body, 3 involved the occipital horn, 2 involved the trigone, 2 involved the frontal horn, and 1 case involved the occipital horn and body. Perioperative complications were analyzed, revealing 1 case of intraoperative acute epidural hematoma intraoperative and 2 cases of postoperative obstructive hydrocephalus. All complications were promptly managed. Postoperative MRI revealed that 14 cases (88%) achieved total resection, while 2 cases (12%) achieved subtotal resection. During the follow-up of 6-38 months, no recurrence was observed. The patient diagnosed with glioblastoma died 16 months after surgery (GOS=1), while the remaining patients have successfully resumed to normal daily life with a GOS score of 5.ConclusionIn conclusion, endoport-assisted neuroendoscopic surgery proved to be a minimally invasive and effective technique for resecting lateral ventricular tumors, with acceptable complications. It effectively utilizes the benefits of close observation, comprehensive exposure, and reduced tissue damage. Therefore, endoport-assisted neuroendoscopic surgery is suitable for the resection of lateral ventricular tumors.
Radiotherapy for Non-Hodgkin’s lymphoma with cardiac infiltration: A case report
Authors: Wang Weifeng , Zhang Zhuo , Deng Xiaocong , Gu Anqi , Chen Xianzhao , Cai Yizheng , Zhao Yuting 
Published: October 2023

BackgroundNon-Hodgkin’s lymphoma (NHL) with cardiac infiltration has a poor prognosis. The median OS of patients failing to respond to chemotherapy has been reported to be 1 month vs. 18 months in patients responding to chemotherapy.Case presentationHerein, we reported a case of a 57-year-old male confirmed with diffuse large B-cell lymphoma who received radiation therapy of 150-cGy daily, administered in 30 fractions to the volume of cardiac infiltration, resulting in complete relief. Chemotherapy had no curative effect. The patient was subsequently enrolled in a clinical trial and received oral administration of zanubrutinib 80mg twice daily, after which he achieved complete remission. The progression-free survival was from diagnosis (January 7, 2020) to the follow-up (September 20, 2022), amounting to 32 months.ConclusionProper irradiation dose and timing of treatment can relieve NHL symptoms.