bluuebunny/crossref_metadata_2025_split huggingface dataset:
CO2-based production of phytase from highly stable expression plasmids in Cupriavidus necator H16
Authors: Arhar Simon , Rauter Thomas , Stolterfoht-Stock Holly , Lambauer Vera , Kratzer Regina , Winkler Margit , Karava Marianna , Kourist Robert , Emmerstorfer-Augustin Anita 
Published: January 2024

Abstract
Background
Existing plasmid systems offer a fundamental foundation for gene expression in Cupriavidus necator; however, their applicability is constrained by the limitations of conjugation. Low segregational stabilities and plasmid copy numbers, particularly in the absence of selection pressure, pose challenges. Phytases, recognized for their widespread application as supplements in animal feed to enhance phosphate availability, present an intriguing prospect for heterologous production in C. necator. The establishment of stable, high-copy number plasmid that can be electroporated would support the utilization of C. necator for the production of single-cell protein from CO2.

Results
In this study, we introduce a novel class of expression plasmids specifically designed for electroporation. These plasmids contain partitioning systems to boost segregation stability, eliminating the need for selection pressure. As a proof of concept, we successfully produced Escherichia coli derived AppA phytase in C. necator H16 PHB− 4 using these improved plasmids. Expression was directed by seven distinct promoters, encompassing the constitutive j5 promoter, hydrogenase promoters, and those governing the Calvin-Benson-Bassham cycle. The phytase activities observed in recombinant C. necator H16 strains ranged from 2 to 50 U/mg of total protein, contingent upon the choice of promoter and the mode of cell cultivation - heterotrophic or autotrophic. Further, an upscaling experiment conducted in a 1 l fed-batch gas fermentation system resulted in the attainment of the theoretical biomass. Phytase activity reached levels of up to 22 U/ml.

Conclusion
The new expression system presented in this study offers a highly efficient platform for protein production and a wide array of synthetic biology applications. It incorporates robust promoters that exhibit either constitutive activity or can be selectively activated when cells transition from heterotrophic to autotrophic growth. This versatility makes it a powerful tool for tailored gene expression. Moreover, the potential to generate active phytases within C. necator H16 holds promising implications for the valorization of CO2 in the feed industry.
Methanol bioconversion into C3, C4, and C5 platform chemicals by the yeast Ogataea polymorpha
Authors: Wefelmeier Katrin , Schmitz Simone , Kösters Benjamin Jonas , Liebal Ulf Winfried , Blank Lars Mathias 
Published: January 2024

Abstract
Background
One carbon (C1) molecules such as methanol have the potential to become sustainable feedstocks for biotechnological processes, as they can be derived from CO2 and green hydrogen, without the need for arable land. Therefore, we investigated the suitability of the methylotrophic yeast Ogataea polymorpha as a potential production organism for platform chemicals derived from methanol. We selected acetone, malate, and isoprene as industrially relevant products to demonstrate the production of compounds with 3, 4, or 5 carbon atoms, respectively.

Results
We successfully engineered O. polymorpha for the production of all three molecules and demonstrated their production using methanol as carbon source. We showed that the metabolism of O. polymorpha is well suited to produce malate as a product and demonstrated that the introduction of an efficient malate transporter is essential for malate production from methanol. Through optimization of the cultivation conditions in shake flasks, which included pH regulation and constant substrate feeding, we were able to achieve a maximum titer of 13 g/L malate with a production rate of 3.3 g/L/d using methanol as carbon source. We further demonstrated the production of acetone and isoprene as additional heterologous products in O. polymorpha, with maximum titers of 13.6 mg/L and 4.4 mg/L, respectively.

Conclusion
These findings highlight how O. polymorpha has the potential to be applied as a versatile cell factory and contribute to the limited knowledge on how methylotrophic yeasts can be used for the production of low molecular weight biochemicals from methanol. Thus, this study can serve as a point of reference for future metabolic engineering in O. polymorpha and process optimization efforts to boost the production of platform chemicals from renewable C1 carbon sources.
Abnormal arginine synthesis confers worse prognosis in patients with middle third gastric cancer
Authors: Hong Lianlian , Tang Xi , Han Jing , Wang Jiaqi , Xu Qianqian , Zhu Xin 
Published: January 2024

Abstract
Background
Gastric cancer at different locations has distinct prognoses and biological behaviors, but the specific mechanism is unclear.

Methods
Non-targeted metabolomics was performed to examine the differential metabolite phenotypes that may be associated with the effects of tumor location on the prognosis of gastric cancer. And silencing of the rate-limiting enzyme to evaluate the effect of abnormal changes in metabolic pathway on the functional biological assays of gastric cancer cells HGC-27 and MKN28.

Results
In a retrospective study of 94 gastric cancer patients, the average survival time of patients with gastric cancer in the middle third of the stomach was significantly lower than that of patients with gastric cancer in other locations (p < 0.05). The middle third location was also found to be an independent risk factor for poor prognosis (HR = 2.723, 95%CI 1.334–5.520), which was closely associated with larger tumors in this location. Non-targeted metabolomic analysis showed that the differential metabolites affected 16 signaling pathways including arginine synthesis, retrograde endocannabinoid signaling, arginine biosynthesis, and alanine and aspartate and glutamate metabolism between gastric cancer and normal tissue, as well as between tumors located in the middle third of the stomach and other locations. Argininosuccinate synthetase 1 (ASS1), the rate-limiting enzyme of the arginine biosynthesis pathway, catalyzes the production of argininosuccinic acid. Here, knockdown of ASS1 significantly inhibited the proliferation, colony formation, and migration/invasion of gastric cancer cells, and promoted apoptosis.

Conclusions
Our study suggests that abnormal arginine synthesis may lead to larger tumor size and worse prognosis in gastric cancer located in the middle third position of the stomach. These findings may provide the basis for the stratification and targeted treatment of gastric cancer in different locations.
The association between hyperuricemia and insulin resistance surrogates, dietary- and lifestyle insulin resistance indices in an Iranian population: MASHAD cohort study
Authors: Seifi Najmeh , Nosrati Mina , Koochackpoor Glareh , Aghasizadeh Malihe , Bahari Hossein , Namdar Hedyeh Beheshti , Afkhami Nafiseh , Darban Reza Assaran , Azarian Farnoosh , Ferns Gordon A. , Ghayour-Mobarhan Majid 
Published: January 2024

Abstract
Background
Previous studies have reported insulin resistance (IR) to be associated with hyperuricemia. In this study, we aimed to assess the possible associations between the empirical dietary index for IR (EDIR), the empirical lifestyle index for IR (ELIR), and non-insulin-based surrogates (triglyceride–glucose (TyG) index, triglyceride-to-high-density-lipoprotein-cholesterol (TG/HDL-C) ratio, metabolic score for insulin resistance (METS-IR) and TyG with body mass index (TyG-BMI)) and hyperuricemia in an Iranian population.

Methods
In this cross-sectional study, 6457 participants aged 35–65 years were recruited as part of the MASHAD cohort study. EDIR and ELIR were calculated using dietary intakes, body mass index, and physical activity information. Insulin resistance surrogates including TyG, TyG-BMI, TG/HDL-C, and METS-IR were calculated for all participants. Hyperuricemia was defined as serum uric acid ≥ 7 mg/dl in men or ≥ 6 mg/dl in women. Multivariable logistic regression models were applied to determine the association between indexes of IR and hyperuricemia.

Results
The mean ELIR and IR surrogates (TyG, TyG-BMI, TG/ HDL, and METS-IR) were significantly higher in subjects with hyperuricemia compared to non-hyperuricemic subjects (p < 0.001). After adjusting for confounding variables, the association between hyperuricemia and EDIR was not significant, but ELIR had a significant association in all models (p < 0.001). All four IR surrogates (TyG, TyG-BMI, TG/ HDL, and METS-IR) showed a significant association with hyperuricemia (p < 0.001).

Conclusion
There was a significant association between indexes of insulin resistance: TyG, TyG-BMI, TG/HDL-c, METS-IR, and ELIR with hyperuricemia, in a population sample from northeastern Iran.
Effect of nutrition education integrating the health belief model and theory of planned behavior on dietary diversity of pregnant women in Southeast Ethiopia: a cluster randomized controlled trial
Authors: Beressa Girma , Whiting Susan J , Belachew Tefera 
Published: January 2024

Abstract
Background
Maternal anemia, miscarriage, low birth weight (LBW), preterm birth (PTB), intrauterine growth restriction (IUGR), prenatal and infant mortality, morbidity, and the risk of chronic disease later in life are all increased by a lack of dietary diversity during pregnancy. However, evidence for the effect of nutrition education on the dietary diversity score (DDS) among pregnant women was sparse in Ethiopia, particularly in the study areas. This study aimed to assess the effect of nutrition education on dietary diversity among pregnant women in urban settings in Southeast Ethiopia.

Methods
A community-based two-arm parallel cluster randomized controlled trial was conducted among 447 randomly selected pregnant women attending antenatal care (224 intervention group and 223 control group) at health facilities from February to December 2021. A multistage cluster sampling technique, followed by systematic sampling, was used to select the pregnant women. Pregnant women who participated in the interventions were given nutrition education starting at 16 weeks of gestation and continuing for 6 months. We used a pre-tested, interviewer-administered, structured questionnaire to collect the data. A 24-hour qualitative dietary recall was used to calculate the dietary diversity score (DDS). A multivariable generalized estimating equation (GEE) model was conducted to evaluate the intervention effect.

Results
After the intervention, the proportion of adequate dietary diversity was 14.15% higher in the intervention arm compared to the control group (45.09% versus 30.94%, P = 0.002). The overall difference in adequate dietary diversity between the two groups was 8.5%. After adjusting for background characteristics, the multivariable GEE binary logistic model revealed that having received intervention [(AOR = 1.89, 95% CI: 1.27, 2.79)], being literate [(AOR = 3.41, 95% CI: 1.13, 10.23)], and having high wealth [(AOR = 1.60, 95% CI: 1.09, 2.35)] significantly improved adequate dietary diversity.

Conclusion
The findings indicated that having received the intervention, being literate, and having a high level of wealth significantly improved maternal dietary diversity. Efforts should be made to increase nutrition education using the health belief model (HBM) and the theory of planned behavior (TPB). Moreover, there is a need to improve literacy and economic empowerment through income-generating activities to enhance adequate dietary diversification during pregnancy.

Trial registration
Clinicaltrials.gov (PACTR202201731802989, Retrospectively registered on 24 January 2022).
The short-term effect of ambient particulate matter on suicide death
Authors: Go Tae-Hwa , Kim Min-Hyuk , Choi Yoon-Young , Han Jaehyun , Kim Changsoo , Kang Dae Ryong 
Published: January 2024

Abstract
Background
Air pollution can cause various respiratory and neurological diseases and continuous exposure can lead to death. Previous studies have reported that particulate matter (PM) exposure increases the risk of depression, suicidal thoughts, and suicidal death; however, the results are inconsistent and limited. This study aimed to examine the relationship between short-term PM exposure and suicide deaths, as well as investigate the short-term effects of PM on suicide death within vulnerable groups based on factors such as sex, age group, suicide-related information (note, method, and cause), psychiatric disorders, and physical diseases.

Methods
Data on a total of 28,670 suicide deaths from 2013 to 2017, provided by the Korea Foundation for Suicide Prevention, were analyzed. The study design employed a time-series analysis with a two-stage approach. In the first step, a generalized additive model combined with a distributed lag nonlinear model was used to estimate the short-term effect of PM exposure on suicide risk specific to each city. In the second step, the estimated results from each city were pooled through a meta-analysis to derive the overall effect. We determined the effects of single lag, cumulative lag, and moving average PM concentrations from days 0–7 before suicide.

Results
We confirmed an association between exposure to PM10 (≤ 10 μm in diameter) and deaths due to suicide. In particular, among individuals with psychiatric disorders and those who employed non-violent suicide methods, increased exposure to PM10 was associated with a higher risk of death by suicide, with percentage changes of 5.92 (95% confidence interval [CI]: 3.95–7.92) and 11.47 (95% CI: 7.95–15.11), respectively. Furthermore, in the group with psychiatric disorders, there was an observed tendency of increasing suicide risk as PM10 levels increased up to 120 µg/m3, whereas in the group with non-violent suicide deaths, there was a pronounced trend of rapid increase in suicide risk with an increase in PM10 up to 100 µg/m3.

Conclusions
These results show an association between short-term exposure to PM and suicide. Our study adds evidence for the benefits of reducing PM in preventing diseases and improving mental health.
Prevalence estimation of Tropheryma whipplei in duodenal biopsy tissues of Koreans
Authors: Yoon Sumi , Hong Min Eui , Hong Soon Auck , Kim Tae-Hyoung , Lee Mi-Kyung 
Published: January 2024

AbstractWhipple’s disease caused by Tropheryma whipplei is difficult to diagnose because of a broad spectrum of manifestations and non-specific clinical signs. In the current global era, the incidence of duodenal infection/inflammation caused by T. whipplei in Korea may has been underestimated. Here we estimated the prevalence of T. whipplei in duodenal biopsy tissues of Koreans using real-time PCRs (RT-PCRs). A total of 252 duodenal biopsy tissues were collected from Korean patients who underwent esophagogastroduodenoscopy and duodenal biopsy. DNA extracted from the duodenal biopsy tissues was analyzed using three RT-PCRs targeting T. whipplei-specific regions of the 16S–23S rRNA intergenic spacer, hsp65, and Dig15 in parallel. In the samples positive in RT-PCRs, direct sequencing was performed for each RT-PCR target. The prevalence of T. whipplei was estimated based on the RT-PCR and sequencing results. Among the analyzed samples, T. whipplei was not detected. The prevalence of T. whipplei in duodenal biopsy tissues of Koreans was estimated to be less than 0.4%. This is the first study to attempt to detect T. whipplei in duodenal biopsy tissues of Koreans and estimate its prevalence. Our findings infer that while T. whipplei carriers exist in Korea, the incidence of duodenal infection/inflammation caused by T. whipplei is extremely rare.
Ultrasound-activated prodrug-loaded liposome for efficient cancer targeting therapy without chemotherapy-induced side effects
Authors: Jiang Yifan , Chen Hongjian , Lin Tao , Zhang Chao , Shen Jiaxin , Chen Jifan , Zhao Yanan , Xu Wen , Wang Guowei , Huang Pintong 
Published: January 2024

Abstract
Background
Off-targeted distribution of chemotherapeutic drugs causes severe side effects, further leading to poor prognosis and patient compliance. Ligand/receptor-mediated targeted drug delivery can improve drug accumulation in the tumor but it always attenuated by protein corona barriers.

Results
To address these problems, a radically different strategy is proposed that can leave the off-targeted drugs inactive but activate the tumor-distributed drugs for cancer-targeting therapy in a tumor microenvironment-independent manner. The feasibility and effectiveness of this strategy is demonstrated by developing an ultrasound (US)-activated prodrug-loaded liposome (CPBSN38L) comprising the sonosensitizer chlorin e6 (Ce6)-modified lipids and the prodrug of pinacol boronic ester-conjugated SN38 (PBSN38). Once CPBSN38L is accumulated in the tumor and internalized into the cancer cells, under US irradiation, the sonosensitizer Ce6 rapidly induces extensive production of intracellular reactive oxygen species (ROS), thereby initiating a cascade amplified ROS-responsive activation of PBSN38 to release the active SN38 for inducing cell apoptosis. If some of the injected CPBSN38L is distributed into normal tissues, the inactive PBSN38 exerts no pharmacological activity on normal cells. CPBSN38L exhibited strong anticancer activity in multiple murine tumor models of colon adenocarcinoma and hepatocellular carcinoma with no chemotherapy-induced side effects, compared with the standard first-line anticancer drugs irinotecan and topotecan.

Conclusions
This study established a side-effect-evitable, universal, and feasible strategy for cancer-targeting therapy.

Graphical Abstract
Thrombolytic therapy based on lyophilized platelet-derived nanocarriers for ischemic stroke
Authors: Migliavacca Martina , Correa-Paz Clara , Pérez-Mato María , Bielawski Patrick-Brian , Zhang Issan , Marie Pauline , Hervella Pablo , Rubio Marina , Maysinger Dusica , Vivien Denis , del Pino Pablo , Pelaz Beatriz , Polo Ester , Campos Francisco 
Published: January 2024

Abstract
Background
Intravenous administration of fibrinolytic drugs, such as recombinant tissue plasminogen activator (rtPA) is the standard treatment of acute thrombotic diseases. However, current fibrinolytics exhibit limited clinical efficacy because of their short plasma half-lives and risk of hemorrhagic transformations. Platelet membrane-based nanocarriers have received increasing attention for ischemic stroke therapies, as they have natural thrombus-targeting activity, can prolong half-life of the fibrinolytic therapy, and reduce side effects. In this study we have gone further in developing platelet-derived nanocarriers (defined as cellsomes) to encapsulate and protect rtPA from degradation. Following lyophilization and characterization, their formulation properties, biocompatibility, therapeutic effect, and risk of hemorrhages were later investigated in a thromboembolic model of stroke in mice.

Results
Cellsomes of 200 nm size and loaded with rtPA were generated from membrane fragments of human platelets. The lyophilization process did not influence the nanocarrier size distribution, morphology, and colloidal stability conferring particle preservation and long-term storage. Encapsulated rtPA in cellsomes and administered as a single bolus showed to be as effective as a continuous clinical perfusion of free rtPA at equal concentration, without increasing the risk of hemorrhagic transformations or provoking an inflammatory response.

Conclusions
This study provides evidence for the safe and effective use of lyophilized biomimetic platelet-derived nanomedicine for precise thrombolytic treatment of acute ischemic stroke. In addition, this new nanoformulation could simplify the clinical use of rtPA as a single bolus, being easier and less time-consuming in an emergency setting than a treatment perfusion, particularly in stroke patients. We have successfully addressed one of the main barriers to drug application and commercialization, the long-term storage of nanomedicines, overcoming the potential chemical and physical instabilities of nanomedicines when stored in an aqueous buffer.
Enhancing immune responses of ESC-based TAA cancer vaccines with a novel OMV delivery system
Authors: Jin Meiling , Huo Da , Sun Jingjing , Hu Jingchu , Liu Shuzhen , Zhan Mingshuo , Zhang Bao-zhong , Huang Jian-Dong 
Published: January 2024

AbstractEmbryonic stem cell (ESC)-derived epitopes can act as therapeutic tumor vaccines against different types of tumors Jin (Adv Healthc Mater 2023). However, these epitopes have poor immunogenicity and stimulate insufficient CD8+ T cell responses, which motivated us to develop a new method to deliver and enhance their effectiveness. Bacterial outer membrane vesicles (OMVs) can serve as immunoadjuvants and act as a delivery vector for tumor antigens. In the current study, we engineered a new OMV platform for the co-delivery of ESC-derived tumor antigens and immune checkpoint inhibitors (PD-L1 antibody). An engineered Staphylococcal Protein A (SpA) was created to non-specifically bind to anti-PD-L1 antibody. SpyCatcher (SpC) and SpA were fused into the cell outer membrane protein OmpA to capture SpyTag-attached peptides and PD-L1 antibody, respectively. The modified OMV was able to efficiently conjugate with ESC-derived TAAs and PD-L1 antibody (SpC-OMVs + SpT-peptides + anti-PD-L1), increasing the residence time of TAAs in the body. The results showed that the combination therapy of ESC-based TAAs and PD-L1 antibody delivered by OMV had significant inhibitory effects in mouse tumor model. Specifically, it was effective in reducing tumor growth by enhancing IFN-γ-CD8+ T cell responses and increasing the number of CD8+ memory cells and antigen-specific T cells. Overall, the new OMV delivery system is a versatile platform that can enhance the immune responses of ESC-based TAA cancer vaccines.
Graphical Abstract
Natural lipid nanoparticles extracted from Morus nigra L. leaves for targeted treatment of hepatocellular carcinoma via the oral route
Authors: Gao Qiang , Chen Nanxi , Li Baoyi , Zu Menghang , Ma Ya , Xu Haiting , Zhu Zhenhua , Reis Rui L. , Kundu Subhas C. , Xiao Bo 
Published: January 2024

AbstractThe clinical application of conventional medications for hepatocellular carcinoma treatment has been severely restricted by their adverse effects and unsatisfactory therapeutic effectiveness. Inspired by the concept of ‘medicine food homology’, we extracted and purified natural exosome-like lipid nanoparticles (LNPs) from black mulberry (Morus nigra L.) leaves. The obtained MLNPs possessed a desirable hydrodynamic particle size (162.1 nm), a uniform size distribution (polydispersity index = 0.025), and a negative surface charge (-26.6 mv). These natural LNPs were rich in glycolipids, functional proteins, and active small molecules (e.g., rutin and quercetin 3-O-glucoside). In vitro experiments revealed that MLNPs were preferentially internalized by liver tumor cell lines via galactose receptor-mediated endocytosis, increased intracellular oxidative stress, and triggered mitochondrial damage, resulting in suppressing the viability, migration, and invasion of these cells. Importantly, in vivo investigations suggested that oral MLNPs entered into the circulatory system mainly through the jejunum and colon, and they exhibited negligible adverse effects and superior anti-liver tumor outcomes through direct tumor killing and intestinal microbiota modulation. These findings collectively demonstrate the potential of MLNPs as a natural, safe, and robust nanomedicine for oral treatment of hepatocellular carcinoma.
Graphic abstract