bluuebunny/crossref_metadata_2025_split huggingface dataset:
RNAscope in situ hybridization reveals microvascular sequestration of Plasmodium relictum pSGS1 blood stages but absence of exo-erythrocytic dormant stages during latent infection of Serinus canaria
Authors: Himmel Tanja , Harl Josef , Matt Julia , Nedorost Nora , Iezhova Tatjana , Ilgūnas Mikas , Valkiūnas Gediminas , Weissenböck Herbert 
Published: March 2024

Abstract
Background
Birds chronically infected with avian malaria parasites often show relapses of parasitaemia after latent stages marked by absence of parasites in the peripheral circulation. These relapses are assumed to result from the activation of dormant exo-erythrocytic stages produced during secondary (post-erythrocytic) merogony of avian Plasmodium spp. Yet, there is no morphological proof of persistent or dormant tissue stages in the avian host during latent infections. This study investigated persistence of Plasmodium relictum pSGS1 in birds with latent infections during winter, with the goal to detect presumed persisting tissue stages using a highly sensitive RNAscope® in situ hybridization technology.

Methods
Fourteen domestic canaries were infected with P. relictum pSGS1 by blood-inoculation in spring, and blood films examined during the first 4 months post infection, and during winter and spring of the following year. After parasitaemia was no longer detectable, half of the birds were dissected, and tissue samples investigated for persisting tissue stages using RNAscope ISH and histology. The remaining birds were blood-checked and dissected after re-appearance of parasitaemia, and their tissues equally examined.

Results
Systematic examination of tissues showed no exo-erythrocytic stages in birds exhibiting latent infections by blood-film microscopy, indicating absence of dormant tissue stages in P. relictum pSGS1-infected canaries. Instead, RNAscope ISH revealed rare P. relictum blood stages in capillaries of various tissues and organs, demonstrating persistence of the parasites in the microvasculature. Birds examined after re-appearance of parasitemia showed higher numbers of P. relictum blood stages in both capillaries and larger blood vessels, indicating replication during early spring and re-appearance in the peripheral circulation.

Conclusions
The findings suggest that persistence of P. relictum pSGS1 during latent infection is mediated by continuous low-level erythrocytic merogony and possibly tissue sequestration of infected blood cells. Re-appearance of parasitaemia in spring seems to result from increased erythrocytic merogony, therefore representing recrudescence and not relapse in blood-inoculated canaries. Further, the study highlights strengths and limitations of the RNAscope ISH technology for the detection of rare parasite stages in tissues, providing directions for future research on persistence and tissue sequestration of avian malaria and related haemosporidian parasites.
Genetic fusion of CCL11 to antigens enhances antigenicity in nucleic acid vaccines and eradicates tumor mass through optimizing T-cell response
Authors: Qi Hailong , Sun Zhongjie , Gao Tianle , Yao Yanling , Wang Yu , Li Weiwei , Wang Xudong , Wang Xiaofang , Liu Defang , Jiang Jian-Dong 
Published: March 2024

AbstractNucleic acid vaccines have shown promising potency and efficacy for cancer treatment with robust and specific T-cell responses. Improving the immunogenicity of delivered antigens helps to extend therapeutic efficacy and reduce dose-dependent toxicity. Here, we systematically evaluated chemokine-fused HPV16 E6/E7 antigen to improve the cellular and humoral immune responses induced by nucleotide vaccines in vivo. We found that fusion with different chemokines shifted the nature of the immune response against the antigens. Although a number of chemokines were able to amplify specific CD8 + T-cell or humoral response alone or simultaneously. CCL11 was identified as the most potent chemokine in improving immunogenicity, promoting specific CD8 + T-cell stemness and generating tumor rejection. Fusing CCL11 with E6/E7 antigen as a therapeutic DNA vaccine significantly improved treatment effectiveness and caused eradication of established large tumors in 92% tumor-bearing mice (n = 25). Fusion antigens with CCL11 expanded the TCR diversity of specific T cells and induced the infiltration of activated specific T cells, neutrophils, macrophages and dendritic cells (DCs) into the tumor, which created a comprehensive immune microenvironment lethal to tumor. Combination of the DNA vaccine with anti-CTLA4 treatment further enhanced the therapeutic effect. In addition, CCL11 could also be used for mRNA vaccine design. To summarize, CCL11 might be a potent T cell enhancer against cancer.
Immune aging in annual killifish
Authors: Morabito Gabriele , Ryabova Alina , Valenzano Dario Riccardo 
Published: March 2024

AbstractTurquoise killifish (Nothobranchius furzeri) evolved a naturally short lifespan of about six months and exhibit aging hallmarks that affect multiple organs. These hallmarks include protein aggregation, telomere shortening, cellular senescence, and systemic inflammation. Turquoise killifish possess the full spectrum of vertebrate-specific innate and adaptive immune system. However, during their recent evolutionary history, they lost subsets of mucosal-specific antibody isoforms that are present in other teleosts. As they age, the immune system of turquoise killifish undergoes dramatic cellular and systemic changes. These changes involve increased inflammation, reduced antibody diversity, an increased prevalence of pathogenic microbes in the intestine, and extensive DNA damage in immune progenitor cell clusters. Collectively, the wide array of age-related changes occurring in turquoise killifish suggest that, despite an evolutionary separation spanning hundreds of millions of years, teleosts and mammals share common features of immune system aging. Hence, the spontaneous aging observed in the killifish immune system offers an excellent opportunity for discovering fundamental and conserved aspects associated with immune system aging across vertebrates. Additionally, the species' naturally short lifespan of only a few months, along with its experimental accessibility, offers a robust platform for testing interventions to improve age-related dysfunctions in the whole organism and potentially inform the development of immune-based therapies for human aging-related diseases.
Excess of body weight is associated with accelerated T-cell senescence in hospitalized COVID-19 patients
Authors: Madruga Mailton Prestes , Grun Lucas Kich , Santos Letícya Simone Melo Dos , Friedrich Frederico Orlando , Antunes Douglas Bitencourt , Rocha Marcella Elesbão Fogaça , Silva Pedro Luis , Dorneles Gilson P. , Teixeira Paula Coelho , Oliveira Tiago Franco , Romão Pedro R.T. , Santos Lucas , Moreira José Claudio Fonseca , Michaelsen Vinicius Schenk , Cypel Marcelo , Antunes Marcos Otávio Brum , Jones Marcus Herbert , Barbé-Tuana Florencia María , Bauer Moisés Evandro 
Published: March 2024

Abstract
Background
Several risk factors have been involved in the poor clinical progression of coronavirus disease-19 (COVID-19), including ageing, and obesity. SARS-CoV-2 may compromise lung function through cell damage and paracrine inflammation; and obesity has been associated with premature immunosenescence, microbial translocation, and dysfunctional innate immune responses leading to poor immune response against a range of viruses and bacterial infections. Here, we have comprehensively characterized the immunosenescence, microbial translocation, and immune dysregulation established in hospitalized COVID-19 patients with different degrees of body weight.

Results
Hospitalised COVID-19 patients with overweight and obesity had similarly higher plasma LPS and sCD14 levels than controls (all p < 0.01). Patients with obesity had higher leptin levels than controls. Obesity and overweight patients had similarly higher expansions of classical monocytes and immature natural killer (NK) cells (CD56+CD16−) than controls. In contrast, reduced proportions of intermediate monocytes, mature NK cells (CD56+CD16+), and NKT were found in both groups of patients than controls. As expected, COVID-19 patients had a robust expansion of plasmablasts, contrasting to lower proportions of major T-cell subsets (CD4 + and CD8+) than controls. Concerning T-cell activation, overweight and obese patients had lower proportions of CD4+CD38+ cells than controls. Contrasting changes were reported in CD25+CD127low/neg regulatory T cells, with increased and decreased proportions found in CD4+ and CD8+ T cells, respectively. There were similar proportions of T cells expressing checkpoint inhibitors across all groups. We also investigated distinct stages of T-cell differentiation (early, intermediate, and late-differentiated – TEMRA). The intermediate-differentiated CD4 + T cells and TEMRA cells (CD4+ and CD8+) were expanded in patients compared to controls. Senescent T cells can also express NK receptors (NKG2A/D), and patients had a robust expansion of CD8+CD57+NKG2A+ cells than controls. Unbiased immune profiling further confirmed the expansions of senescent T cells in COVID-19.

Conclusions
These findings suggest that dysregulated immune cells, microbial translocation, and T-cell senescence may partially explain the increased vulnerability to COVID-19 in subjects with excess of body weight.
Food insecurity, environment, institutional quality, and health outcomes: evidence from South Asia
Authors: Azimi Mohammad Naim , Rahman Mohammad Mafizur 
Published: March 2024

Abstract
Background
Food insecurity and environmental degradation pose significant threats to health outcomes in South Asia, necessitating effective policy interventions. Therefore, this study aims to examine the impact of food insecurity and environmental degradation on health outcome indicators amidst global inflationary shocks and institutional quality arrangements. Additionally, it aims to explore the intricate moderating role of institutional quality on the relationship between food insecurity, endogenous variables, and external shocks.

Method
In alignment with the study’s objectives, a set of panel data spanning from 2000 to 2021 is compiled for South Asia. The study introduces a novel variable representing inflationary shock, crafted through the integration of inflation datapoints and the application of the generalized autoregressive conditional heteroskedasticity model. Additionally, a distinctive aggregate institutional quality index is formulated, drawing from six key measures of the Worldwide Governance indicators. To scrutinize the effects of food insecurity, environmental degradation, and other explanatory variables, the study employs the two-step system generalized method of moment technique, offering a robust analytical approach to uncover complex relationships and dynamics in the region.

Results
The results indicate that the prevalence of undernourishment, inequality in per capita calorie intake, and CO2 emissions significantly reduce life expectancy and increase mortality rates. Additionally, it shows that per capita kilocalorie supply, per capita GDP, per capita health expenditures, and urbanization are statistically significant for increasing life expectancy and decreasing mortality rates. The findings reveal that inflationary shocks severely affect food insecurity and environmental factors, exerting further pressure on contemporary life expectancy and mortality rates. In rebuttal, the institutional quality index is found to have significant effects on increasing and decreasing life expectancy and mortality rates, respectively. Furthermore, the institutional quality index is effective in moderating the nexus between food insecurity, environmental degradation, and health outcomes while also neutralizing the negative impact of inflationary shocks on the subject.

Conclusion
The results verify triple health constraints such as food insecurity, environmental factors, and economic vulnerability to global shocks, which impose severe effects on life expectancy and mortality rates. Furthermore, poor institutional quality is identified as a hindrance to health outcomes in South Asia. The findings suggest specific policy implications that are explicitly discussed.
Attractive combinations of female gingival displays, buccal corridor sizes, and facial heights according to orthodontists, dentists, and laypeople of different ages and sexes: a psychometric study
Authors: Niknam Ozra , Yousefi Hafshejani Shakila , Rakhshan Vahid 
Published: March 2024

Abstract
Introduction
Esthetics plays a crucial role in orthodontics and many other dental and medical fields. To date, no study has assessed the combined effects of the 3 facial features ‘facial height, gingival display (GD), and buccal corridor size (BC)’ on facial/smile beauty. Therefore, this study was conducted for the first time.

Methods
In this psychometric diagnostic study, beauty of 27 randomized perceptometric images of a female model with variations in facial heights (short, normal, long), gingival displays (0, 2, 4, 6 mm), and buccal corridor sizes (2%, 10%, 15%, 20%, 25%) were evaluated by 108 judges (36 orthodontists, 36 dentists, 36 laypeople) using a 5-scale Likert scale (1 to 5). Combined effects of facial heights, GDs, BCs, judges’ sexes, ages, and jobs, and their 2-way interactions were tested using a mixed-model multiple linear regression and a Bonferroni test. Zones of ideal features were determined for all judges and also for each group using repeated-measures ANOVAs and the Bonferroni test (α=0.05).

Results
Judges’ sex but not their age or expertise might affect their perception of female beauty: men gave higher scores. The normal face was perceived as more beautiful than the long face (the short face being the least attractive). Zero GD was the most attractive followed by 4 mm; 6 mm was the least appealing. BCs of 15% followed by 10% were the most attractive ones, while 25% BC was the worst. The zone of ideal anatomy was: long face + 0mm GD + 15% BC; normal face + 2mm GD + 15% BC; long face + 2mm GD + 15% BC; normal face + 0mm GD + 15% BC.

Conclusions
Normal faces, zero GDs, and 15% BCs may be the most appealing. Facial heights affect the perception of beauty towards GDs but not BCs.
Detection of HER2 expression using 99mTc-NM-02 nanobody in patients with breast cancer: a non-randomized, non-blinded clinical trial
Authors: Zhao Lingzhou , Xing Yan , Liu Changcun , Ma Shaofei , Huang Wenhua , Cheng Zhen , Zhao Jinhua 
Published: March 2024

Abstract
Background
99mTc radiolabeled nanobody NM-02 (99mTc-NM-02) is a novel single photon emission computed tomography (SPECT) probe with a high affinity and specificity for human epidermal growth factor receptor 2 (HER2). In this study, a clinical imaging trial was conducted to investigate the relationship between 99mTc-NM-02 uptake and HER2 expression in patients with breast cancer.

Methods
Thirty patients with pathologically confirmed breast cancer were recruited and imaged with both 99mTc-NM-02 SPECT/computed tomography (CT) and 18F-fluorodeoxyglucose (18F-FDG) positron emission tomography (PET)/CT. According to the treatment conditions before recruitment, patients were divided into two groups, the newly diagnosed group (n = 24) and the treated group (n = 6). The maximal standard uptake value (SUVmax) of 18F-FDG and SUVmax and mean SUV (SUVmean) of 99mTc-NM-02 in the lesions were determined to analyze the relationship with HER2 expression.

Results
No meaningful relationship was observed between 18F-FDG uptake and HER2 expression in 30 patients with breast cancer. 99mTc-NM-02 uptake was positively correlated with HER2 expression in the newly diagnosed group, but no correlation was observed in the treated group. 99mTc-NM-02 uptake in HER2-positive lesions was lower in those with effective HER2-targeted therapy compared with the newly diagnosed group. 99mTc-NM-02 SPECT/CT detected brain and bone metastases of breast cancer with a different imaging pattern from 18F-FDG PET/CT. 99mTc-NM-02 showed no non-specific uptake in inflamed tissues and revealed intra- and intertumoral HER2 heterogeneity by SPECT/CT imaging in 9 of the 30 patients with breast cancer.

Conclusions
99mTc-NM-02 SPECT/CT has the potential for visualizing whole-body HER2 overexpression in untreated patients, making it a promising method for HER2 assessment in patients with breast cancer.

Trial registration
NCT04674722, Date of registration: December 19, 2020.
Joint analysis of mutational and transcriptional landscapes in human cancer reveals key perturbations during cancer evolution
Authors: Cho Jae-Won , Cao Jingyi , Hemberg Martin 
Published: March 2024

Abstract
Background
Tumors are able to acquire new capabilities, including traits such as drug resistance and metastasis that are associated with unfavorable clinical outcomes. Single-cell technologies have made it possible to study both mutational and transcriptomic profiles, but as most studies have been conducted on model systems, little is known about cancer evolution in human patients. Hence, a better understanding of cancer evolution could have important implications for treatment strategies.

Results
Here, we analyze cancer evolution and clonal selection by jointly considering mutational and transcriptomic profiles of single cells acquired from tumor biopsies from 49 lung cancer samples and 51 samples with chronic myeloid leukemia. Comparing the two profiles, we find that each clone is associated with a preferred transcriptional state. For metastasis and drug resistance, we find that the number of mutations affecting related genes increases as the clone evolves, while changes in gene expression profiles are limited. Surprisingly, we find that mutations affecting ligand-receptor interactions with the tumor microenvironment frequently emerge as clones acquire drug resistance.

Conclusions
Our results show that lung cancer and chronic myeloid leukemia maintain a high clonal and transcriptional diversity, and we find little evidence in favor of clonal sweeps. This suggests that for these cancers selection based solely on growth rate is unlikely to be the dominating driving force during cancer evolution.
Multi-omics reveals that alkaline mineral water improves the respiratory health and growth performance of transported calves
Authors: Qi Jiancheng , Gan Linli , Huang Fangyuan , Xie Yue , Guo Hongrui , Cui Hengmin , Deng Junliang , Gou Liping , Cai Dongjie , Pan Chunhui , Lu Xia , Shah Ali Mujtaba , Fang Jing , Zuo Zhicai 
Published: March 2024

Abstract
Background
Long-distance transportation, a frequent practice in the cattle industry, stresses calves and results in morbidity, mortality, and growth suppression, leading to welfare concerns and economic losses. Alkaline mineral water (AMW) is an electrolyte additive containing multiple mineral elements and shows stress-mitigating effects on humans and bovines.

Results
Here, we monitored the respiratory health status and growth performance of 60 Simmental calves subjected to 30 hours of road transportation using a clinical scoring system. Within the three days of commingling before the transportation and 30 days after the transportation, calves in the AMW group (n = 30) were supplied with AMW, while calves in the Control group (n = 29) were not. On three specific days, namely the day before transportation (day -3), the 30th day (day 30), and the 60th day (day 60) after transportation, sets of venous blood, serum, and nasopharyngeal swab samples were collected from 20 calves (10 from each group) for routine blood testing, whole blood transcriptomic sequencing, serology detection, serum untargeted metabolic sequencing, and 16S rRNA gene sequencing. The field data showed that calves in the AMW group displayed lower rectal temperatures (38.967 ℃ vs. 39.022 ℃; p = 0.004), respiratory scores (0.079 vs. 0.144; p < 0.001), appetite scores (0.024 vs. 0.055; p < 0.001), ocular and ear scores (0.185 vs. 0.338; p < 0.001), nasal discharge scores (0.143 vs. 0.241; p < 0.001), and higher body weight gains (30.870 kg vs. 7.552 kg; p < 0.001). The outcomes of laboratory and high throughput sequencing data revealed that the calves in the AMW group demonstrated higher cellular and humoral immunities, antioxidant capacities, lower inflammatory levels, and intestinal absorption and lipogenesis on days -3 and 60. The nasopharynx 16S rRNA gene microbiome analysis revealed the different composition and structure of the nasopharyngeal microflora in the two groups of calves on day 30. Joint analysis of multi-omics revealed that on days -3 and 30, bile secretion was a shared pathway enriched by differentially expressed genes and metabolites, and there were strong correlations between the differentially expressed metabolites and the main genera in the nasopharynx.

Conclusions
These results suggest that AMW supplementation enhances peripheral immunity, nutrition absorption, and metabolic processes, subsequently affecting the nasopharyngeal microbiota and improving the respiratory health and growth performance of transported calves. This investigation provided a practical approach to mitigate transportation stress and explored its underlying mechanisms, which are beneficial for the development of the livestock industry.
Ectopic epipericardial fat necrosis: a case report
Authors: Yoshino Ryusei , Nakatsubo Masaki , Ujiie Nanami , Ito Akane , Yoshida Nana , Aoki Naoko , Kitada Masahiro 
Published: March 2024

Abstract
Background
Epipericardial fat necrosis (EFN) is a rare disease in which local inflammation and necrosis occur in the adipose tissue surrounding the heart, particularly epicardial fat. Few cases of EFN in which surgical resection was performed have been reported. We report a case of EFN after surgical resection of a right extrapulmonary tumor, in which a malignant disease could not be excluded.

Case presentation
A 75-year-old male patient presented with fever and chest pain. A contrast-enhanced computed tomography scan of the chest revealed a lesion, 53 × 48 mm in size, with mixed fatty density spanning the middle and lower lobes of the right lung. Thoracic magnetic resonance imaging (MRI) revealed a mass with mixed fat and soft tissue density in the same area; the lesion was contiguous with pericardial fatty tissue. The tumor was diagnosed as a liposarcoma or teratocarcinoma based on imaging results; however, the possibility of lung cancer could not be excluded. Finally, EFN was diagnosed based on the postoperative histopathological examination. The patient underwent surgical resection of the suspected right extrapulmonary tumor. The intraoperative findings revealed a mediastinal mass contiguous with pericardial fat located between the middle and lower lobes. Intraoperative pathological examination of the lesion was performed using a needle biopsy; however, no definitive diagnosis was made. The tumor may have invaded the middle lobe of the right lung, and partial resection of the right lower lobe was performed in addition to resection of the middle lobe of the right lung. The patient was followed up every 3 months without adjuvant therapy. No recurrence was reported at 1 year after surgery.

Conclusion
EFN should be considered in the differential diagnosis of an extrapulmonary tumor when continuity with the pericardial space is observed on MRI or other imaging studies. Surgical resection is useful in the diagnosis and treatment of EFNs. Preoperative three-dimensional reconstructive imaging and MRI should be used to identify vascular structures and confirm the continuity of the lesion with the surrounding tissues to ensure safe and rapid tumor removal.