Dietary Alaska Pollock Protein Attenuates the Experimental Colitis Induced by Dextran Sulfate Sodium via Regulation of Gut Microbiota and Its Metabolites in Mice

Protein derived from fish has not only nutritional properties but also health-promoting properties. Few studies have examined the effect of dietary Alaska pollock protein (APP) on the anticolitis effect reported to be associated with metabolic syndrome (MetS). This study investigated the effect of APP intake on colitis symptoms, gut microbiota, and its metabolites in the experimental colitis mouse model induced by dextran sulfate sodium (DSS). Male C57BL/6J mice were divided into three groups: (1) DSS-untreated mice fed an American Institute of Nutrition (AIN) 93G diet (protein source is casein), (2) DSS-treated mice fed an AIN93G diet, and (3) DSS-treated mice fed an APP diet. After the mice were fed the diets for 21 days, experimental colitis was induced by three cycles of 2% DSS administration for 5 days followed by washouts over the course of 5 days. APP-reduced body weight loss increased the disease activity index, and elevated spleen weight and alleviated colon length shortening and colonic tissue damage. Furthermore, APP altered the structure and composition of the microbiota and short-chain fatty acids in feces. Since APP intake alleviates experimental colitis induced by DSS administration through alterations in the gut microbiota and its metabolites, we deduced that APP would inhibit MetS progression via colitis suppression.

Sphingosine-1-Phosphate and Its Signal Modulators Alleviate Psoriasis-Like Dermatitis: Preclinical and Clinical Evidence and Possible Mechanisms

BackgroundPsoriasis is an autoimmune skin disease associated with lipid metabolism. Sphingosine-1-phosphate (S1P) is a bioactive lipid that plays a key role in the development of autoimmune diseases. However, there is currently a lack of comprehensive evidence of the effectiveness of S1P on psoriasis.ObjectiveTo assess the efficacy and possible mechanism of S1P and its signal modulators in the treatment of psoriasis-like dermatitis.MethodsSix databases were searched through May 8, 2021, for studies reporting S1P and its signal modulators. Two reviewers independently extracted information from the enrolled studies. Methodological quality was assessed using SYRCLE’s risk of bias tool. RevMan 5.3 software was used to analyze the data. For clinical studies, the Psoriasis Area and Severity Index score were the main outcomes. For preclinical studies, we clarified the role of S1P and its regulators in psoriasis in terms of phenotype and mechanism.ResultsOne randomized double-blind placebo-controlled trial and nine animal studies were included in this study. The pooled results showed that compared with control treatment, S1P receptor agonists [mean difference (MD): −6.80; 95% confidence interval (CI): −8.23 to −5.38; p<0.00001], and sphingosine kinase 2 inhibitors (MD: −0.95; 95% CI: −1.26 to −0.65; p<0.00001) alleviated psoriasis-like dermatitis in mice. The mechanism of S1P receptor agonists in treating psoriasis might be related to a decrease in the number of white blood cells, topical lymph node weight, interleukin-23 mRNA levels, and percentage of CD3+ T cells (p<0.05). Sphingosine kinase 2 inhibitors ameliorated psoriasis in mice, possibly by reducing spleen weight and cell numbers (p<0.05).ConclusionsS1P receptor agonists and sphingosine kinase 2 inhibitors could be potential methods for treating psoriasis by decreasing immune responses and inflammatory factors.

A Cadaveric Study on Morphometric Features of Spleen and Splenomegaly with Accessory Spleen in Hilum

Background: Anatomical knowledge regarding the external morphology of the spleen is essential for surgical intervention and radiological diagnosis. Splenomegaly is defined as pathologic enlargement of the spleen measured by size or weight. A normal spleen has a craniocaudal length of no more than 12 cm and weighs less than 200 g. It is surrounded by a thin capsule. The spleen is usually not palpable unless it is enlarged; therefore, a palpable spleen is almost always abnormal. At times the spleen may be difficult to palpate, but dullness to percussion during inspiration in the area of the lower left intercostal space in the left anterior axillary line suggests splenic enlargement. Massive splenomegaly, weight >1000 g usually occurs in lymphoma, myeloproliferative disorders, visceral leishmaniasis, and malaria. Materials and Methods: This study was conducted in different medical institutions, to find morphometric features, spenomegaly in cadaver during routine anatomy dissection as part of curriculum, 100 cadavers were observed to find out splenomegaly. Results: Out of 100 spleens studied, 81 cases wedge shaped spleen was the most common, followed by 12 tetrahedral shaped spleens and 7 oval shaped spleens. Average weight of the spleen was 175g. Average length of the spleen was 11.64cm, Average breadth of the spleen was 7.3cm and average thickness of spleen was 3.6cm. Out of 100 cadavers observed only one cadaver observed with massive splenomegaly with one accessory spleen in hilum. The spleen weight was 875gm, length was 18.15 cm, width was 8.65cm, thickness was 5.75cm and extended upto 7 rib and it is easily palpable below the rib cage from lumbar aspect. The cadaver was male and age around 55 years. Conclusion: The morphometric knowledge of spleen will helpful for surgeons and for understanding deceases related spleen. The knowledge of splenomegaly is important in finding splenic disorders and accessory spleen information helpful in understanding embryonic development of spleen. KEY WORDS: Splenomegaly, Spleen, Hilum of Spleen, Accessory spleen.

RhANP attenuates endotoxin-derived cognitive dysfunction through subdiaphragmatic vagus nerve-mediated gut microbiota–brain axis

Background Atrial natriuretic peptide (ANP) secreted from atrial myocytes is shown to possess anti-inflammatory, anti-oxidant and immunomodulatory effects. The aim of this study is to assess the effect of ANP on bacterial lipopolysaccharide (LPS)-induced endotoxemia-derived neuroinflammation and cognitive impairment. Methods LPS (5 mg/kg) was given intraperitoneally to mice. Recombinant human ANP (rhANP) (1.0 mg/kg) was injected intravenously 24 h before and/or 10 min after LPS injection. Subdiaphragmatic vagotomy (SDV) was performed 14 days before LPS injection or 28 days before fecal microbiota transplantation (FMT). ANA-12 (0.5 mg/kg) was administrated intraperitoneally 30 min prior to rhANP treatment. Results LPS (5.0 mg/kg) induced remarkable splenomegaly and an increase in the plasma cytokines at 24 h after LPS injection. There were positive correlations between spleen weight and plasma cytokines levels. LPS also led to increased protein levels of ionized calcium-binding adaptor molecule (iba)-1, cytokines and inducible nitric oxide synthase (iNOS) in the hippocampus. LPS impaired the natural and learned behavior, as demonstrated by an increase in the latency to eat the food in the buried food test and a decrease in the number of entries and duration in the novel arm in the Y maze test. Combined prophylactic and therapeutic treatment with rhANP reversed LPS-induced splenomegaly, hippocampal and peripheral inflammation as well as cognitive impairment. However, rhANP could not further enhance the protective effects of SDV on hippocampal and peripheral inflammation. We further found that PGF mice transplanted with fecal bacteria from rhANP-treated endotoxemia mice alleviated the decreased protein levels of hippocampal polyclonal phosphorylated tyrosine kinase receptor B (p-TrkB), brain-derived neurotrophic factor (BDNF) and cognitive impairment, which was abolished by SDV. Moreover, TrkB/BDNF signaling inhibitor ANA-12 abolished the improving effects of rhANP on LPS-induced cognitive impairment. Conclusions Our results suggest that rhANP could mitigate LPS-induced hippocampal inflammation and cognitive dysfunction through subdiaphragmatic vagus nerve-mediated gut microbiota–brain axis.

Aging Predisposes B cells to Malignancy by Activating c-Myc and Perturbing the Genome and Epigenome

Age is the single major risk factor for human cancer, but naturally occurring cancers are rarely studied in aging models. Like humans, mice spontaneously develop cancer with age, and standard laboratory strains are predisposed for B-cell lymphoma. Here, we uncover how B-cell lymphoma develops as a consequence of the aging immune system. We found that aged B cells acquire somatic mutations in tumor suppressors and oncogenes (e.g. Trp53, Pim1, and Myh11) and undergo monoclonal expansions, with some clones representing 86% of splenic B cells. Clonal B cells had hypermethylated promoters and globally silenced expression, suggesting a role of DNA methylation in clonal selection of premalignant B cells. B-cell size, spleen weight, and a novel population of B cells, which we named Myc+ cells, emerged as convenient markers of malignancy. High-throughput analyses of clonal B cells and the use of genetic mouse models revealed that c-Myc drives B-cell size increase and clonal expansion with age. Phoshoproteome and co-culture experiments revealed that c-Myc is activated by signals from the aging microenvironment. Moreover, single-cell RNA-seq suggested that clonal B cells originate from age-associated B cells, further underlying the importance of aging environment in cancer transformation. Longitudinal analyses demonstrated a negative impact of premalignant B cells on mouse lifespan and linked it to age-related myeloid bias. Together, our study revealed cell-autonomous changes that cooperate with the aging microenvironment to give rise to preneoplastic B cells. This stidy established a novel model to study how aging predisposes cells to cancer transformation.

Effect of supplementing feed grade L-methionine in plant sourced diet on the performance of broiler chicken

The study was conducted to investigate the productivity and profitability of broiler on L-methionine supplemented diet. A total of 160 day-old broiler chicks (Cobb 500) was randomly distributed to four dietary supplements, D0 (Control), D1 (0.30 % L-Met), D2 (0.35 % L- Met) and D3 (0.40 % L- Met). Each treatment was replicated five times with 8 birds per cage. The chicks were reared in the battery cages while they were fed with broiler starter diet (crumble) until Day14 of age. The birds with fed with L-Met supplemented diets starting from Day15 until Day35 of age All the diets were iso-caloric and iso-nitrogenous.. Results revealed that the L-Met supplementations were in favor of higher live weight (LW) and better (p<0.05) feed conversion ratio without affecting the feed intake (FI). The D3 diet resulted higher (p<0.01) LW in the birds than all others on Day35 of age. The livability (%) was unaffected (p>0.05) by the dietary supplements. No significant (p>0.05) difference was observed in the relative weights of visceral organs (small intestine, proventriculus, gizzard, liver and heart) except for the spleen. Relatively increased (p<0.05) spleen weight (%) was observed in the D3 diet. The breast weight (%) was increased marginally (p<0.07) in the D3 group, while the other carcass parts; breast weight, thigh weight, wing weight, shank weight, drumstick weight, neck weight, and abdominal fat content were found to be similar (p>0.05) in the dietary groups. Higher profit (p<0.01) and lower production cost (p<0.05) were observed in the D3 group than that of the others. It can be inferred that 0.40% L-Met supplementation in the broiler diet is better for growth performance and profitability. Bang. J. Livs. Res. Vol. 27 (1&2), 2020: P. 1-14

Mitapivat Improves Transfusion Burden and Reduces Iron Overload in Thalassemic Mice

Mitapivat, an oral activator of pyruvate kinase (PK), was recently shown to improve b-thalassemic anemia with a reduction of ineffective erythropoiesis and an amelioration of b-thalassemic red cell features in a mouse model for b-thalassemia (Hbb 3th/+ mice).). These changes were also associated with a beneficial effect on iron homeostasis by modulation of duodenal DMT1 expression (Matte A et al JCI 2021). Two clinical studies have shown improvement of anemia and ineffective erythropoiesis with mitapivat treatment in patients with non-transfusion-dependent (NTD) thalassemia (Kuo et al. EHA 2021). Based on these results, Phase 3 studies in both NTD and TD thalassemia are currently on going. The objective of this preclinical study was to determine if treatment with mitapivat affects the length between transfusion of red blood cells (RBCs) and the liver iron concentration (LIC). Using a previously established murine model of RBCs transfusions (Park Y et al Blood 2020), in Hbb 3th/+ mice, we used Hb 10.5 g/dL as threshold for RBCs transfusion, with washed RBCs, at 40% Hct (400 uL total volume infused). The animals were divided into two groups: vehicle and mitapivat (50mg/Kg by gavage BID for up to 61 days).The length of the interval between transfusions increased in mitapivat treated compared to vehicle treated animals (transfusion interval: 13.8±1.0 days vs vehicle 10.5±1.0 days respectively n=4 and n=3). In both groups, the transfusion regimen induced a significant reduction in spleen weight/mouse weight ratio and in extramedullary erythropoiesis. We also found a significant reduction in liver iron content (LIC) in mitapivat treated compared to vehicle treated animals. We then evaluated the effects of mitapivat in combination with iron chelation using deferiprone (DFP,1.25 mg/mL, drinking water). Casu et al. have previously shown in the same mouse model for β-thal that DFP did not affect erythropoiesis. In the β-thal mice, we did not find negative effects on hematologic parameters when mitapivat (50 mg/Kg/d by gavage BID) was co-administrated with DFP for 28 days. LIC was reduced in mitapivat treated mice and in mitapivat +DFP treated β-thal mice was further decreased compared to vehicle treated animals. This allowed us to reduce DFP dosage from 1.25 to 0.8 mg/mL in mitapivat treated β-thal mice. These data show that in mouse model of transfused β-thalassemia, mitapivat increases the time interval between transfusions, reduces transfusion burden and allows a reduction of the dosing iron chelation with DFP. Thus, mitapivat might represent an interesting option in transfusion dependent β-thalassemic patients, being transfusion burden still an unmet need in this patient population. Disclosures Kosinski: Agios Pharmaceuticals: Current Employment, Current equity holder in publicly-traded company. Dang: Agios Pharmaceuticals, Inc.: Current Employment, Current holder of stock options in a privately-held company. De Franceschi: F. Hoffmann-La Roche Ltd: Consultancy.

Metabolomics Based on 1H-NMR Reveal the Regulatory Mechanisms of Dietary Methionine Restriction on Splenic Metabolic Dysfunction in Obese Mice

Methionine restriction (MR) has been reported to have many beneficial health effects, including stress resistance enhancement and lifespan extension. However, the effects of MR on the splenic metabolic dysfunction induced by obesity in mice remain unknown. This study aimed to investigate the scientific problem and clarify its possible mechanisms. C57BL/6J mice in the control group were fed a control diet (0.86% methionine, 4.2% fat) for 34 weeks, and others were fed a high-fat diet (0.86% methionine, 24% fat) for 10 weeks to establish diet-induced obese (DIO) mouse models. Then, the obtained DIO mice were randomly divided into two groups: the DIO group (DIO diet), the DIO + MR group (0.17% methionine, 24% fat) for 24 weeks. Our results indicated that MR decreased spleen weight, and spleen and plasma lipid profiles, promoted lipid catabolism and fatty acid oxidation, glycolysis and tricarboxylic acid cycle metabolism, and improved mitochondrial function and ATP generation in the spleen. Moreover, MR normalized the splenic redox state and inflammation-related metabolite levels, and increased plasma levels of immunoglobulins. Furthermore, MR increased percent lean mass and splenic crude protein levels, activated the autophagy pathway and elevated nucleotide synthesis to maintain protein synthesis in the spleen. These findings indicate that MR can ameliorate metabolic dysfunction by reducing lipid accumulation, oxidative stress, and inflammation in the spleen, and the mechanism may be the activation of autophagy pathway.

Large-Scale Multivariate Analysis to Interrogate an Animal Model of Stroke: Novel Insights Into Poststroke Pathology

Background and Purpose: Preclinical stroke studies endeavor to model the pathophysiology of clinical stroke, assessing a range of parameters of injury and impairment. However, poststroke pathology is complex and variable, and associations between diverse parameters may be difficult to identify within the usual small study designs that focus on infarct size. Methods: We have performed a retrospective large-scale big data analysis of records from 631 C57BL/6 mice of either sex in which the middle cerebral artery was occluded by 1 of 5 surgeons either transiently for 1 hour followed by 23-hour reperfusion (transient middle cerebral artery occlusion [MCAO]; n=435) or permanently for 24 hours without reperfusion (permanent MCAO; n=196). Analyses included a multivariate linear mixed model with random intercept for different surgeons as a random effect to reduce type I and type II errors and a generalized ordinal regression model for ordinal data when random effects are low. Results: Analyses indicated that brain edema volume was associated with infarct volume at 24 hours (β, 0.52 [95% CI, 0.45–0.59]) and was higher after permanent MCAO than after transient MCAO ( P <0.05). A more severe clinical score was associated with a greater infarct volume but not with the animal’s age or edema volume. Further, a more severe clinical score was observed for a given brain infarct volume after transient MCAO versus permanent MCAO. Remarkably the animal’s age, which corresponded with the period of young adulthood (6–40 weeks; equivalent to ≈18–35 years in humans), was positively associated with severity of lung infection (β, 0.65 [95% CI, 0.42–0.88]) and negatively with spleen weight (β, −0.36 [95% CI, −0.63 to −0.09]). Conclusions: Large-scale analysis of preclinical stroke data can provide researchers in our field with insight into relationships between variables not possible if individual studies are analyzed in isolation and has identified hypotheses for future study.

PSI-B-26 Association of leptin gene polymorphisms (C73T and C528T) on the development of internal organs in Aberdeen-angus cattle

Numerous studies have established the relationship of SNPs in the leptin gene with fat and energy exchange, nutritional behavior in cattle. In turn, the intensity of metabolic processes in body is determined by the development of internal organs. The purpose of research was to study the relationship between the complex of polymorphisms C73T and C528T in leptin gene for the formation of internal organs in Aberdeen-Angus cows and heifers. Cows (n = 30) and heifers (n = 49) were genotyped and identified respectively 7 and 8 various combinations of genotypes of C73T (GenBank AF120500) and C528T (GenBank AB070368). There were no homozygous CC/CC and TT/TT combinations in cows, and no TT/TT combination in heifers. Cows were slaughtered at the age of 3 years, and heifers - at 20 months. Cows with haplotype CT/CC had the highest liver, heart, lung, kidney and spleen weight with superiority relative to peers of 6.5–16.0% (P &gt; 0.05). The minimum development of stomachs was established at cows with a combination of genotypes of CC/TT which conceded to analogs for 7.6–13.8% (P &gt; 0.05). Haplotypes CT/CT and TT/CT in heifers were associated with greater internal organ massiveness. Carriers of complex CT/CT genotype outperformed peers in terms of liver, heart, lung, kidney and spleen weight by 1.75–6.59% (P &lt; 0.05, P &gt; 0.05), and individuals with a combination of TT/CT by 3.50–8.84% (P &lt; 0.05, P &gt; 0.05). Less developed stomachs were established in bodies with haplotype TT/CC, which were inferior to other genotypes by 1.20–7.49% (P &lt; 0.05, P &gt; 0.05). This indirectly indicates a relatively weak ability to use coarse plant foods. Thus, the complex genotype inleptin gene is associated with the variability in the development of internal organs, which determines the intensity of metabolic processes. This research was performed with financial support from the project of RAS0526-2021-0001.