bluuebunny/crossref_metadata_2025_split huggingface dataset:
Milk protein and fat in Saanen goats
Authors: Mudruk Semen , Bakhta Alesia , Karpenko Larisa , Kozitcyna Anna , Polistovskaia Polina 
Published: May 2022

Dairy industry is one of the leading branches of animal husbandry. Dairy goat breeding is becoming more and more relevant each day; it has already acquired special significance for the agro‐industry of Russian Federation. In addition to the outstanding milk productivity of Saanen goats, it is worth noting the versatility of the milk processing, which is important for farms additionally engaged in manufacturing of goat cheese and other dairy products. Therefore, the chemical composition of goat milk depending on various physiological states of goats is of study interest.Research was conducted in North‐Western region of Russian Federation, in Saanen goats. Five groups included 30 Saanen goats each, from 1stto 5th lactations. The milk samples were taken once during control milking. Levels of milk fat and proteins were assessed. Results presented as mean±standard error of the mean. Student's t‐test was used after proving normal distribution. Level of significance is labeled as p<0.05.As a result of presented study, it was revealed that, in goats of 1st lactation the milk protein level was 3.1±0.22% and milk fat was 3.8±0.98%, in animals of 2nd lactation the milk protein level was 2.9±0.21% and fat was 3.7±0.52%, in goats of 3rd lactation the protein level was 2.8±0.23% and milk fat was 3.3±0.58%, in animals of 4th lactation the milk protein level was 2.1±0.23% and fat was 3.3±0.53%, in animals of 5th lactation, the protein level was 3.1±0.27% and milk fat was 3.5±0.48%. The average values according to these indicators were 3.0±0.24% for milk protein, 3.6±0.73% for milk fat. When analyzing the milk protein obtained results level he minimum values were noted in goats of 4th lactation. This marker was significantly (p<0.05) lower by 30% relative to the average protein content in milk. The maximum amount of milk protein was observed in goats of 1st and 5th lactations. There were no significant differences in animals of 1st, 2nd, 3rd and 5th lactations when compared to the average value for the entire lactation period. When analyzing the amount of milk fat it was noted that there were no significant differences in the goats of 1st, 2nd, 3rd and 4th lactations. There was a tendency to milk fat decreasing in animals of 3rd, 4thand 5th lactations relative to the average values for the entire lactation period, and a tendency to milk fat increasing in goats of 1st and 2nd lactations by 5.5% and 2.77%, respectively.Consequently this data is important to be taken into consideration keeping goats and planning further processing of milk depending on the prevalent number of presented goats in the herd.
Nuclear Location Bias of HCAR1 Drives Cancer Malignancy by Multiple Routes
Authors: Mohammad Nezhady Mohammad Ali , Cagnone Gael , Chemtob Sylvain 
Published: May 2022

IntroductionG‐Protein Coupled Receptors (GPCR) are virtually involved in all physiological processes. HCAR1 (GPR81), as a GPCR, is endogenously activated by lactate and has been shown to promote cancer malignancy via higher level of glycolysis due to Warburg effect. Its expression level is highly elevated in many cancers and negatively correlates with patient’s prognosis. However, its mechanism of action is not well understood.On the other hand, nuclear localization of several GPCRs have been described albeit it is unusual feature for them. Additionally, it has been shown that nuclear GPCRs can perform functions distinct from their cell surface counterparts in vivo. Here we demonstrate HCAR1 has a nuclear localization and this localization pattern promotes cancer malignancy by multiple routes.Methods and ResultsWe determined HCAR1 nuclear localization pattern by cell fractionation, immunofluorescence confocal imaging and TEM. Site‐directed mutagenesis showed ICL3 and phosphorylation of C‐terminal domains are required for nuclear localization. We also demonstrated that this localization is ligand independent and there is a pool of nuclear HCAR1 (N‐HCAR1) in the cells. We show N‐HCAR1 induces intra‐nuclear signaling through Gi and Gßγ by WB and ELISA leading to increased phosphorylation of nuclear AKT and ERK resulting in increased cancer cell survival and proliferation. Our ChIP‐sequencing data shows N‐HCAR1 binds to the genes regulating transcription and promoting expression of genes involved in cell migration and we validated this in cellulo proving N‐HCAR1 promotes migration. We identified N‐HCAR1 interactome by Bio‐ID coupled with mass spectrometry and found, it interacts with proteins involved in ribosome biogenesis, translation and DNA damage repair and our experimental data demonstrates that specifically the N‐HCAR1 promotes these three process in cellulo. Additionally, we identified the transcriptomic signature of N‐HCAR1 and showed it regulated a larger gene network than its plasma membrane counterpart. Concordantly, our in vivo tumor xenografts and tail vein injections proves that tumors without N‐HCAR1 have smaller size and tumor mass and lower metastatic rate as well.ConclusionHere we show an unusual localization of a GPCR in the nucleus and provide evidence that N‐HCAR1 specifically promotes various hallmarks of cancer malignancy. The effect of N‐HCAR1 is validate in vivo in tumor xenografts as well. Understanding these mechanisms can provide targets and cues for therapeutic developments.
Gut Microbial and Metabolic Signatures are Altered in Adolescents with Type 1 Diabetes
Authors: Pon Velayutham Anandh Babu , Mokhtari Pari , Metos Julie M. , Jambal Puujee , Shankar Kartik 
Published: May 2022

BackgroundEvidence suggests that the pathogenesis and development of Type 1 Diabetes (T1D) are driven by both genetic predisposition and immune dysfunction which may be influenced by gut microbiota. The present study aimed to investigate the composition and functional pathways of the gut microbiome in adolescents with T1D, determine the association between gut microbes and clinical and dietary factors in T1D, and identify features predictive of T1D.MethodsHealthy adolescents (n=35) and adolescents with T1D (n=30) were recruited and microbial profiling in participants’ stool samples was performed using deep shotgun metagenomic sequencing. The bioBakery3 pipeline (Kneaddata, Metaphlan3 and HUMAnN) was used to analyze the reads to assign taxonomy and functional annotations. The differences in taxonomic abundances were identified using Linear Discriminant Analysis (LDA) Effect Size (LEfSe) and ecological measures (diversity) were assessed using standard methods. The association between the abundance of specific bacterial species and the metadata (HbA1C, skin carotenoids, diet and physical activity) was assessed by performing Spearman’s correlation. Random forest modeling (using the MUVR package) was used to identify features predictive of T1D.ResultsAdolescents with T1D exhibited a significant increase in Simpson α‐diversity index as compared to healthy adolescents. β‐diversity was similar between the two groups. LEfSe indicated a significant difference in the microbial community at different taxonomic levels between the two groups. Seven taxa were enriched in adolescents with T1D at the species level that includes Alistipes Shahii, Asaccharobacter celatus, Blautia obeum, Coprococcus eutactus, Coprobacillus cateniformis, Clostridium symbiosum, and Eggerthella lenta. Three species Alistripes Putredins, Dialister invisus, and Eubacterium ventriosum were enriched in healthy adolescents. Further, 33 out of 400 microbial metabolic pathways were significantly different between the two groups. Importantly, biosynthesis of amino acids, vitamins, enzyme cofactors, and electron carrier were downregulated whereas fermentation pathways were upregulated in adolescents with T1D as compared to healthy adolescents. Association analysis indicated that specific bacterial species are correlated with HbA1C, skin carotenoids, dietary factors and physical activity. Most of the bacterial species associated with these measures were different between the two groups. MUVR identified bacterial taxa predictive of T1D status (0.82 sensitivity and 0.63 specificity). The top features in using genus level abundance included Coprococcus, Faecalibaterium and Streptococcus.ConclusionThe present study indicates an extensive alteration in the composition and functional capacity of gut microbiota in adolescents with T1D providing new insight into microbial and metabolic signatures in adolescents with T1D.
Endothelial‐specific nanodelivery of Rho kinase inhibitors: Targeting tumor angiogenesis and metastasis
Authors: Lahooti Behnaz , Akwii Racheal , Mattheolabakis George , Mikelis Constantinos M. 
Published: May 2022

ObjectiveThe limitations of current vascular endothelial growth factor (VEGF)‐centered anti‐angiogenic therapy include the inhibition of physiological angiogenesis, development of evasive resistance, and increased vascular leakage, which drives the need for a novel therapy for tumor angiogenesis and metastasis. RhoA pathway inhibition blocks VEGF‐ and sphingosine‐1 phosphate (S1P)‐induced angiogenesis and vascular leakage without hindering endogenous angiogenesis. Fasudil, a clinically relevant Rho‐associated kinase (ROCK) inhibitor, blocks the RhoA downstream signaling, but adverse events (hypotension, skin reactions) exist. Here, we propose endothelial ROCK targeting with Fasudil‐containing immune‐related nanodelivery.HypothesisFasudil‐encapsulated CD31‐targeting liposomal formulation is a potential tumor antiangiogenic therapy by blocking RhoA signaling via ROCK inhibition in the endothelial cells.MethodsFasudil‐liposomal nanoparticles were prepared by DPPC, cholesterol, 16:0 PEG2000PE, DSPE‐PEG2000‐mal, DSPE‐PEG‐FITC, and CHCl3: MeOH (4:1) combination, thin‐film formation, downsizing after Fasudil incorporation, filtering, and conjugation with IgG and CD31‐targeting antibodies. We used dynamic light scattering (DLS) for the liposomal characterization, dose‐response experiments on primary Human Umbilical Vein Endothelial Cells (HUVEC) and Bend3 cells for in vitro toxicity evaluation, antibody (Ab)‐to‐liposome binding evaluation and pathway inhibition evaluation by western blot, and UPLC analysis (EE)% for the evaluation of encapsulation efficiency. The binding affinity of the CD31‐conjugated liposomes and their time‐dependent cell incorporation were evaluated by competitive binding assay via immunohistochemistry (IHC) and flow cytometry. Morphometric analysis was assessed by Livecyte imaging analysis.ResultsSize and PDI measurement by DLS confirmed the stability of our formulation, which presented limited toxicity (concentration: 0.625‐5120 µM; IC50 at 43.05 µM [HUVECs over 24 hours of treatment]) with a significantly higher overall survival rate than Fasudil. Efficient CD31 and mouse IgG Ab conjugation to Fasudil liposomes and blockade of the RhoA downstream signaling in endothelial cells were confirmed. The specific binding of CD31‐targeting liposomes on endothelial cells was verified in the presence and absence of excess CD31 Ab. The time‐dependent study confirmed the kinetics of liposome incorporation in endothelial cells and the enhanced uptake of CD31 liposomes compared to IgG. Cell morphometric and physiological analysis revealed comparable alterations caused by Fasudil‐ and endothelial‐specific Fasudil‐containing liposomes.ConclusionWe have successfully developed Fasudil‐encapsulated, CD31‐targeting liposomes. Future studies will be directed to study the formulation’s effect on endothelial physiology in vitro and in vivo, tumor growth, angiogenesis, and metastasis.
Loss of PTPN2 Activity Alters Iron Handling Protein Expression in IBD Patients and Causes Iron Deficiency in Mice
Authors: Lei Hillmin , Shawki Ali , Spalinger Marianne R. , Canale Vinicius , Santos Alina N. , Chatterjee Pritha , Crawford Meli’sa S. , Manz Salomon , Becerra Anica , Scharl Michael , McCole Declan F. 
Published: May 2022

BackgroundAnemia is the most common extraintestinal complication of inflammatory bowel disease (IBD) and is a risk factor for Crohn’s disease (CD) onset. Iron deficiency is the most common cause of anemia in IBD; however, the mechanisms involved are poorly understood. Here, we investigated the role of the IBD risk gene, protein tyrosine phosphatase non‐receptor type 2 (PTPN2), in regulating iron homeostasis.MethodsProteomic analyses were performed on serum from IBD patients genotyped for the IBD‐associated loss‐of‐function rs1893217 PTPN2variant (n=10/genotype). Constitutive Ptpn2 wild type (WT), heterozygous (Het), and knockout (KO) 3‐week‐old mice were analyzed for serum iron content, liver non‐heme iron and liver expression of the iron regulatory hormone, hepcidin (Hamp1). Protein and RNA from duodenal epithelial cells (DECs) were assayed by western blotting and qPCR. Localization of the brush border ferrous iron transporter, DMT1, in duodenal tissue was determined by immunohistochemistry (IHC).ResultsIron homeostasis genes, the iron carrier transferrin (TF) and the transferrin receptor (TFRC), were reduced (‐log p‐value = 10.7) in CD patients with the PTPN2 risk variant. Ptpn2‐KO mice had reduced i) serum iron (p<0.001; n=11); ii) serum TF saturation (p<0.01; n=9); and iii) liver non‐heme iron levels (p<0.01; n=10), vs. Ptpn2‐WT and Het mice. This indicated that PTPN2 loss decreased serum and liver iron storage. Moreover, Ptpn2‐KO mice had reduced liver expression of Hamp1 (p<0.01; n=7), likely suppressed by low serum iron. DEC gene expression of ferritin (Fth1), an intracellular iron storage molecule, was reduced (p=0.048; n=5) while Tfrc1, a mediator of basolateral cellular iron uptake, was significantly increased (p=0.0048; n=6) in Ptpn2‐KO mice. Reduced FTH1 expression (p=0.007; n=12) in DECs of Ptpn2‐KO mice was confirmed by western blot, indicating reduced intracellular iron storage. DMT1 expression was unchanged (n=6), whereas IHC showed reduced apical membrane DMT1 in duodenal epithelium of Ptpn2‐KO mice (n=6), suggesting a possible mechanism of impaired intestinal non‐heme iron uptake.ConclusionsCD patients with the PTPN2 loss‐of‐function rs1893217 SNP display alterations in serum iron handling proteins. Loss of PTPN2 activity in mice causes features of anemia including iron deficiency associated with mislocalization of the duodenal transporter DMT1. These findings identify a major role for PTPN2 in regulating iron homeostasis.
Conditional deletion of the glucocorticoid receptor in vascular smooth muscle disrupts the circadian rhythm of vascular reactivity and reduces blood pressure
Authors: Krilis Georgios , Bailey Matthew A. , Ivy Jessica R. 
Published: May 2022

Glucocorticoids synchronise peripheral clocks with the master clock in the suprachiasmatic nucleus in response to the ambient light cycle. In humans and mice, arrhythmic glucocorticoids induce non‐dipping blood pressure and vascular dysfunction. The mechanisms of this are poorly understood. We hypothesize that arrhythmic activation of the glucocorticoid receptor attenuates the circadian clock signalling in the vasculature leading to changes in blood pressure. To test this hypothesis, we assessed vascular function and blood pressure rhythms in control mice and mice with smooth muscle specific deletion of the glucocorticoid receptor (SMGRKO).Renal and mesenteric arteries were isolated at 7am (ZT0; lights on) and 7pm (ZT12; lights off), mounted on a wire myograph and the responses to increasing doses of phenylephrine or sodium nitroprusside (SNP) assessed. In wild‐type mice, the sensitivity of the renal artery to phenylephrine‐mediated contraction was greater at ZT12 than at ZT0, by comparing the half maximal effective concentration of each group, (logEC50: 7.54±0.26 versus 7.29±0.17 respectively; p=0.04). The sensitivity to SNP‐mediated relaxation was also greater when measured at ZT12 compared to ZT0 (7.64±0.3 versus 7.09±0.5; p=0.03). Similar responses were also detected in the mesenteric arteries. In SMGRKO mice, the temporal differences in renal and mesenteric artery vascular reactivity were absent. We used radiotelemetry to assess the effect of this altered rhythm of vascular reactivity on blood pressure. SMGRKO had significantly lower systolic blood pressure throughout the 24h cycle compared to wild‐type littermates, both when lights were off (107.9 ± 22.13 versus 136.3 ± 17.17 mmHg respectively; p=0.015) and during the lights on period (99.6 ± 13.16 versus 118 ± 15.9 mmHg; p=0.014). Blood pressure rhythmicity was not altered in SMGRKO mice.These data show that the glucocorticoid receptor signalling in vascular smooth muscle contributes both to the diurnal variation of vascular reactivity and to blood pressure, but loss of the receptor does not affect blood pressure rhythm. Whether glucocorticoid receptor deletion in smooth muscle protects against non‐dipping blood pressure induced by glucocorticoids needs further investigation.
Mapping Spatio‐temporal ADAR Editing Landscapes in Neurodevelopment
Authors: Plonski Noel‐Marie , Meindl Richard , Piontkivska Helen 
Published: May 2022

BackgroundTranscriptome diversity, more specifically RNA editing, plays a fundamental role in neurodevelopment and plasticity. One form of RNA editing, catalyzed by Adenosine Deaminases Acting on RNA (ADAR), creates variant isoforms, many of which are involved in important pathways including differentiation, proliferation, cell‐migration, vesicular trafficking, and receptor endocytosis and recycling. Immune activation can lead to changes in gene expression of one specific isoform, ADAR1p150, altering RNA editing landscapes causing changes to the structure and/or function of proteins, which, in turn, may contribute to the broad spectrum of symptoms seen in many neurological disorders.MethodsUsing publically available RNA‐seq datasets combined with our computational pipeline, Automated Isoform Diversity Detective (AIDD), including our novel statistical approach using Guttman scaling, we examine global ADAR‐editing patterns in different regions and time points during brain development. In addition, to explore the more subtle nuances of editing changes, nucleotide frequencies at individual genes within the excitome are used to determine differential expression of variant isoform.ResultsADAR expression and global ADAR editing patterns are significantly different between both brain regions and gestational week. Additionally, ADAR editing landscapes are spatially and temporally regulated, especially prominent in the excitome, a collection of 109 experimentally confirmed ADAR editing sites. A few editing sites are unique to each developing region, while other sites are differentially edited across gestational weeks. Finally, serotonin receptor 2C (5HTR2C), with 5 well‐studied editing sites, is differentially edited early in development in choroid plexus and later in development in the basal ganglia, but not edited in the cerebellum.ConclusionsMapping the global RNA editing patterns during neurodevelopment offers insights into mechanisms behind the evolution of higher cognitive function, as well as into molecular pathways underpinning neurological disease. Changes in the excitome variant isoform expression can potentially be used as diagnostic, prognostic and/or therapeutic biomarkers in neurodevelopmental, psychiatric, and degenerative disorders.
Functional MYO5B Loss Alters Lipid Metabolic Pathways in Intestinal Progenitor Cells
Authors: Kaji Izumi 
Published: May 2022

Background and Aims: Differentiation of intestinal epithelial cells involves cell division inhibition, cell lineage choice, and brush border elaboration. Myosin Vb (MYO5B) is a motor protein that is critical for cell polarization and membrane protein trafficking in intestinal epithelial cells and its deficiency causes both hyperproliferation and microvillus defects. Recently, we reported that a bioactive phospholipid, lysophosphatidic acid (LPA), can promote cell differentiation and nutrient absorption in tamoxifen‐induced MYO5B knockout mice in vivo and in enteroids. Our RNA‐sequencing data with KEGG pathway and GO enrichment analysis indicated that MYO5B loss disrupts stem cell characteristics, cell lineage differentiation, and energy metabolic pathways. We hypothesize that the alteration of metabolic pathways in progenitor cells causes the disruption of stem cell function.Methods: Adult Villin‐CreERT2;Myo5bflox/flox or littermate control (Myo5bflox/flox) mice received a single dose of tamoxifen and the intestinal tissues were analyzed 4 days after the tamoxifen injection. Fresh frozen sections of jejunum were analyzed by imaging mass spectrometry (IMS). Immunostaining for key metabolic enzymes were utilized to support the RNA‐seq and IMS observations.Results: Using RNA‐sequencing, MYO5B deficient epithelial cells showed significant decreases in transcripts for enzymes that mediate fatty acid ß‐oxidation compared to control mouse jejunum. Instead, Acsl3, Acat2, and Hmgcs2 were significantly upregulated, proteins that are important for lipogenesis, cholesterol ester synthesis, and ketogenesis, respectively. Consistent with these results, IMS demonstrated that MYO5B deficient tissues accumulated long‐chain fatty acids, phosphatidylinositol, cholesterol derivatives, and nucleotide products in the mucosa (Figure 1). Immunostaining signals of HMGCS2 in control mouse jejunum is limited to the crypt stem cells, whereas MYO5B‐deficient intestine showed expanded HMGCS2 expression in the proliferative cell zone. Interestingly, systemic LPA treatment in MYO5B‐deficient mice further increased the expression of HMGCS2 in both proliferating and differentiated epithelial cells (Figure 2). These observations suggest that MYO5B loss induces a starvation‐like phenotype in proliferative epithelial cells and that LPA treatment increased ketone body production as alternative energy fuels, resulting in the promotion of epithelial cell differentiation.Conclusion: MYO5B loss impairs fatty acid oxidation in the intestinal progenitor cells, which likely leads to cell differentiation deficits. Alteration of metabolic pathways might be a therapeutic target for epithelial disfunction seen in diarrheal diseases.