Glycogen Deposition Research Articles

Histochemical Study of Human Placental Tissues in Gestational Diabetic Mellitus

Gestational diabetes mellitus (GDM) is a serious pregnancy complication in which a woman who has never had diabetes develops chronic hyperglycemia during her pregnancy. Normal placental function is essential for optimal fetal growth. The transport of glucose from the mother to the fetus is critical for fetal nutrient demands and can be stored as glycogen in the placenta. However, the function of this glycogen deposition is unknown: It may well be a source of fuel for a placenta itself or the storage reservoir for the later use by the fetus in times of need. While the significance of the placental glycogen remains unknown, the mounting evidence indicates that the altered glycogen metabolism and/or deposition is associated with many pregnancy complications, such as gestational diabetes, that adversely affect fetal development. The aim of this study is to assess glycogen deposition using Histochemical staining of Periodic Acid Schiff (PAS) stain. The placenta tissue collected from 50 women were enrolled in this study (25 women with no complications) and (25 women with gestational diabetes). The placentas of the two groups were compared in this study based on glycogen deposition with periodic acid-Schiff stain. The results of a histochemical investigation using PAS stain revealed a significant increase in the glycogen deposition (p≤0.001) in diabetic women's placentas within the intervillous core, around fetal vessels, and the basement membranes.

Adverse effects of perinatal protein restriction on glucose homeostasis in offspring of Sprague-Dawley rats

Adequate evidence suggests that poor in-utero environment produced by early life dietary disturbance may predispose offspring to chronic diseases in adult life. It remains to be defined the adverse effect of early exposure due to perinatal protein restriction (PPR) (gestation, lactation and/or both) on the regulators of glucose homeostasis of the male offspring in later life. Hence, the present study investigated the role of perinatal protein restriction on the regulators of glucose homeostasis in adult offspring. Twenty-four pregnant and twelve male Sprague-Dawley rats were used. They were fed either a normal diet containing 20% protein or protein-restricted (PR) diet with 8% protein. The dams were given PR diet up to parturition (in-utero group, IUPR), or from birth to postnatal day 21 (lactation group, LPR) or for a period covering both groups (combined group, CPR). Control dams with control diet was run in parallel for comparison. At day 120, oral glucose tolerance, insulin tolerance, fasting blood glucose, fasting insulin level were determined. Hepatic and myocytic glycogen contents, GLUT-4 protein abundance, alpha amylase and alpha glucosidase activities were also assessed. The results revealed that glucose tolerance was unchanged, insulin sensitivity was significantly decreased in IUPR and CPR offspring. HOMA-IR elicited a significant elevation (p<0.01) in IUPR and CPR offspring compared with CONT. Hepatic and myocytic glycogen contents in all PPR offspring significantly decreased compared with CONT. GLUT-4 protein was significantly (p<0.01) reduced in all PPR offspring compared with CONT. Alpha amylase activity decreased significantly (p<0.01) in IUPR and CPR offspring with a significant increase (p<0.01) in LPR offspring compared with CONT. However, alpha glucosidase activity showed no significant difference (p>0.01) in all PPR offspring compared with CONT. In conclusion, the data described in the present investigation provides ample evidence that PPR remarkably malprogram the offspring to impairment of glucose homeostasis with blunted insulin action in the tissues of the periphery which eventually reduces uptake of glucose, utilization and absorption as well as deposition of glycogen in liver and skeletal muscle leading to insulin resistance in PPR offspring.

Goose nephritic astrovirus infection increases autophagy, destroys intercellular junctions in renal tubular epithelial cells, and damages podocytes in the kidneys of infected goslings

Goose nephritic astrovirus (GNAstV) has recently been identified, which causes kidney swelling and visceral gout in goslings. However, the pathological changes in kidney tissue due to GNAstV infection have not yet been described. In the study, fifty goslings were orally infected with GNAstV, and fifty goslings received PBS as a control. Kidney tissue was collected at different days following infection (dpi) to assess the injury. GNAstV infection reduced body weight, increased the relative weight of the kidney, and increased serum uric acid and creatinine levels. GNAstV was found within renal epithelial cells, and the viral load in the kidney peaked at 7 dpi. Pale and swollen kidney tissue was observed in infected goslings, especially at 5 and 7 dpi. GNAstV infection caused degeneration and necrosis of renal epithelial cells, structural destruction of the brush border, glycogen deposition in the glomerular mesangium, increased fibrosis, and infiltration of inflammatory cells into the renal interstitium. Moreover, swollen mitochondria, broken mitochondrial ridges, autophagosomes, and autophagolysosomes were observed under ultrahistopathological examination. GNAstV infection increased levels of LC3B, ATG5, and Beclin 1, and decreased p62, and downregulated WT1 mRNA and upregulated desmin mRNA. At early stages, GNAstV infection decreased expression of intercellular junction-related genes, including ZO-1, occludin, claudin-10, and catenin-α2. In conclusion, GNAstV infection causes renal epithelial cell autophagy, destruction of brush border and intercellular junctions, podocyte damage, and increased fibrosis, ultimately resulting in damage to the kidney.

S2596 Glycogenic Hepatopathy: A Rare Cause of Reversible Severe Hepatitis

Introduction: Glycogenic hepatopathy (GH) is a rare reversible complication of poorly controlled diabetes mellitus. Its presentation can range from mild to severe hepatitis and diagnosis can be challenging. Here we present a case of GH presenting as severe hepatitis. Case Description/Methods: A 51-year-old male with a history of hypertension and poorly uncontrolled T1DM presented with two days of intractable nausea and vomiting. Physical examination revealed cachectic male only oriented to name. Abdominal exam was normal without tenderness to palpation or organomegaly. Laboratory results were significant for bicarbonate < 5.0 mmol/L, glucose 1169 mg/dL, total bilirubin (T bili.) 0.4 mg/dL, alkaline phosphatase (ALKP) 141 U/L, alanine aminotransferase (ALT) 161 U/L, aspartate aminotransferase (AST) 107 U/L, albumin 3.6 g/dL, INR 1.0, hemoglobin A1c 13.7%. Head computed tomography showed no acute intracranial pathology. He was admitted to the intensive care unit, diagnosed with DKA and treated with insulin infusion therapy. On day 4, laboratory results showed T bili. 0.4 mg/dL, ALKP 295 U/L, ALT 1570 U/L, AST 5250 U/L, INR 1.0. An ultrasound showed mild hepatomegaly 14cm, hyperechoic liver parenchyma, no intra or extra hepatic biliary dilation, and patent hepatic and portal veins. Workup of viral hepatitis A, B and C, autoimmune hepatitis, Wilson’s disease, alpha 1 antitrypsin deficiency, and hemochromatosis was negative. An ultrasound guided liver biopsy showed diffuse swollen hepatocytes, minimal focal fibrosis and mild inflammation composed of scattered lymphocytes, plasma cells and few polymorphonuclear leukocytes. Periodic acid-Schiff (PAS) stain with and without diastase highlighted abundant cytoplasmic glycogen. Patient was diagnosed with GH and a rapid downtrend in liver enzymes was noted with DKA treatment. Discussion: GH was first described by Peter Mauriac in 1930 in children with uncontrolled T1DM. Its incidence is unknown as there have been less than 100 cases reported so far in English literature. Pathophysiology of GH remains unclear but rapid fluctuations in glucose and insulin leading to excessive glycogen deposition in hepatocytes has been suggested. Distinguishing GH from NASH, DILI and other chronic liver disease is challenging without a liver biopsy. With proper glycemic control, GH is quickly reversible and has a good prognosis. No long term hepatic complications are known. GH remains underdiagnosed due to a combination of diagnostic challenges and lack of awareness amongst specialists.Figure 1.: Histopathological section of the liver showing: A) Hepatocytes with clear and fine granular cytoplasm under hematoxylin and eosin stain, B) PAS stain showing large amount of dark red-colored glycogen in hepatocytes, C) Complete digestion of glycogen after adding diastase stain and reappearance of clear hepatocytes.

Network pharmacology-based investigation of potential targets of astragalus membranaceous-angelica sinensis compound acting on diabetic nephropathy

To explore the mechanism of the Astragalus membranaceous (AM)-Angelica sinensis (AS) compound in the treatment of diabetic nephropathy (DN) we used network pharmacology and molecular docking. Screen the components and targets of the AM-AS compound in the TCMSP and the BATMAN-TCM, and establish a component-target interaction network by Cytoscape 3.7.2. After searching relevant targets of DN in related databases, the common targets of the AM-AS compound and DN were obtained by comparison. Gene ontology (GO) analysis and Kyoto Encyclopedia of Gene and Genome (KEGG) pathway enrichment analysis were performed through David database. Molecular docking was performed by PyMoL2.3.0 and AutoDock Vina software. After screening, 142 main targets of the AM-AS compound in the treatment of DN have been identified. Target network was established and the topology of PPI network was analyzed. KEGG pathway enrichment analysis shows that these targets are related to apoptosis, oxidative stress, inflammation, insulin resistance, etc. Molecular docking shows that the target proteins have good combinations with the main active components of the AM-AS compound. AM-AS compound may treat DN by acting on VEGFA, TP53, IL-6, TNF, MARK1, etc., and regulate apoptosis, oxidative stress, inflammation, glucose, and lipid metabolism processes. The in vivo study results suggest that AM-AS compound can significantly reduce the FBG level of diabetic rats, increase the level of INS, improve renal functions, reduce urinary proteins, inhibit glycogen deposition, granulocyte infiltration and collagen fiber proliferation in renal tissue, and restrain the progress of DN. In vivo study combined with network pharmacology and molecular docking methods provides new ideas for the pathogenesis and treatments of DN.

STUDY THE HARMFUL EFFECTS OF DIETHYL PHTHALATE ON LIVER AND TESTES OF MALE MICE, (Mus musculus)

The current study was designed in order to evaluate the harmful effects of Diethyl Phthalate (DEP) on testes and liver of 8 weeks-old mouse Mus musculus, which were grouped as Control group and Experimental groups. Control group was given corn oil and experimental groups were given dose of DEP I 1.2 mg/g B.W and DEP II 1.60 mg/g B.W in super refined corn oil with the help of syringe for 21 days. Dose quantity was 0.1 ml. Glutathione Peroxidase 5(GPx5) and Malondialdehyde (MDA) hormones were used. Morphological examination was done on both experimental group and control group. Methods In Dose group, DEP I (1.2 mg/g B W) various deformities were observed i.e. wrinkled surface, shrinkage and reduced size of testes, while in dose group, DEP II (1.60 mg/g BW) showed abnormalities in their size, appearance , shape, their surface become wrinkled and shrinkage of cell also observed. In Morphometric analysis the control and experimental group showed significant results regarding length, width and weight. In control group there is no certain change in the weight, length and width of testes of male mice, whereas in dose group, DEP 1 (1.2 mg/g B.W) and DEP II (1.60 mg/g B.W) shows the significant effects on decreasing length, width and weight of testes in both groups. In the current experiment, the glutathione level has decreased after the intake of dose in male mice. Experimental group show decreased level of glutathione while control group show the normal level. In the recent experiment the level of Malondialdehyde (MDA) significantly increased after administration of certain doses. In control group, MDA level remains normal while in the experimental group, MDA level increases and the antioxidant enzyme of testicles, glutathione peroxidase (GPx) and a hormone testosterone significantly decreased. Liver histological examination in control group show normal deposition of glycogen, normal and intact association of sinusoid, portal vein, with complete lobular boundary, and appear

Maternal choline supplementation in a rat model of periconceptional alcohol exposure: Impacts on the fetus and placenta.

Maternal choline supplementation in rats can ameliorate specific neurological and behavioral abnormalities caused by alcohol exposure during pregnancy. We tested whether choline supplementation ameliorates fetal growth restriction and molecular changes in the placenta associated with periconceptional ethanol exposure (PCE) in the rat. Sprague Dawley dams were given either 12.5% ethanol (PCE) or 0% ethanol (Con) in a liquid diet from 4days prior to 4days after conception. At day 5 of pregnancy, dams were either placed on a standard chow (1.6g choline/kg chow) or an intermediate chow (2.6g choline/kg chow). On day 10 of pregnancy, a subset of the intermediate dams were placed on a chow further supplemented with choline (7.2g choline/kg chow), resulting in 6groups. Fetuses and placentas were collected on day 20 of pregnancy for analysis. Choline supplementation resulted in increased fetal weight at late gestation, ameliorating the deficits due to PCE. This was most pronounced in litters on a standard chow during pregnancy. Choline also increased fetal liver weight and decreased fetal brain:liver ratio, independent of alcohol exposure. Placental weight was reduced as choline levels in the chow increased, particularly in female placentas. This resulted in a greater ratio of fetal:placental weight, suggesting increased placental efficiency. Global DNA methylation in the placenta was altered in a sex-specific manner by both PCE and choline. However, the increased glycogen deposition in female placentas, previously reported in this PCE model, was not prevented by choline supplementation. Our results suggest that choline has the potential to ameliorate fetal growth restriction associated with PCE and improve placental efficiency following prenatal alcohol exposure. Our study highlights the importance of maternal nutrition in moderating the severity of adverse fetal and placental outcomes that may arise from prenatal alcohol exposure around the time of conception.

P–385 The relationship between systemic oestradiol and vaginal microbiota composition in miscarriage and normal pregnancy

Abstract Study question Is there an association between serum oestradiol, vaginal microbial composition and pregnancy outcome in the early first trimester? Summary answer In women with a vaginal microbiome deplete of Lactobacillus species at the time of Pregnancy of Uncertain Viability (IPUV), higher serum oestradiol associates with livebirth. What is known already During pregnancy, oestradiol mediates vaginal mucosal properties and increases glycogen deposition in epithelial cells which is thought to support colonisation of Lactobacillus species. Low levels of Lactobacillus associates with adverse outcomes such as miscarriage and preterm birth. The direct relationship between systemic oestradiol and the vaginal microbiome has never been studied in pregnancy. However studies have shown a positive correlation between serum oestrone, vaginal glycogen and Lactobacillus abundance in menopausal women. Study design, size, duration This was a prospective cohort study where one-hundred women were recruited in early pregnancy at the time of IPUV and donated paired blood and vaginal samples. 40 women had an eventual miscarriage, 58 had a livebirth and two pregnancies were terminated. All 100 women donated one paired serum and vaginal sample at this time point, and 22 women with Lactobacillus depletion at the time of IPUV donated further longitudinal vaginal samples. Participants/materials, setting, methods Participants were recruited from an Early Pregnancy Unit and underwent transvaginal ultrasound assessment of their pregnancy. Serum samples were analysed with an immunoassay on a ROCHE COBAS E411 analyser for Oestradiol (pg/ml) and Progesterone (ng/ml). Bacterial DNA was extracted from paired vaginal swabs and sequenced using Illumina MiSeq sequencing of 16S rRNA gene amplicons. Main results and the role of chance Lactobacillus dominance of the vagina was associated with higher serum levels of E2 and progesterone compared to depletion (E2=398pg/ml vs 302pg/ml(p = 0.02), P4=23.1ng/ml vs 17ng/ml(p = 0.02)). E2 and P4 were positively correlated (r = 0.6, p &amp;lt; 0.05). At species level, L. crispatus dominance associated with significantly higher levels of E2 compared to high-diversity communities (468pg/ml vs 302pg/ml(p = 0.03) but no such relationship was observed for P4. Both E2 and P4 levels were lower in women who eventually miscarried. However there was no significant difference in the vaginal bacterial composition at genera or species level at this early gestational age (P = 0.08) regardless of per vaginal bleeding. However in women with Lactobacillus depleted microbiota, livebirth was associated with significantly higher E2 levels compared to women suffering miscarriage (212pg/ml in miscarriage vs 395pg/ml in livebirth, p = 0.003) (OR = 22.4 P = 0.004). In 22 women who had Lactobacillus depletion at the time of IPUV (7 with an eventual outcome of miscarriage, and 15 with an eventual outcome of livebirth), longitudinal vaginal bacterial DNA sequencing was performed. In 7/15 women with livebirth, and higher E2 levels, the microbial composition changed to become more Lactobacillus dominant during pregnancy, whereas in those with miscarriage, only 1/7 changed to become Lactobacillus dominant. Limitations, reasons for caution In this study, serum oestradiol levels were compared to the local vaginal bacterial environment. The ideal would be to study local vaginal oestradiol, glycogen and the bacterial composition. Wider implications of the findings: In contrast to previous studies in menopause where low oestrogen levels associate with the vaginal microbial composition, this study uses the high oestradiol environment of early pregnancy to study the mechanistic relationship between oestradiol and vaginal Lactobacillus abundance. Trial registration number NA

Contemporary Approaches to Physiological Adaptations after Taper Workouts

This study was conducted to systematically compile and sythesize the studies about taper training in literature and in the most current form, to reveal the physiological changes caused by taper trainings. Qualitative research methods were used for in-depth study and interpretation of the studies on taper applications published between 1985-2020. Document analysis was used as data collection method and the obtained data were analyzed by content analysis method. Taper training is a complex training method that facilitates the systematic reduction of the training load and the attainment of the physiological harmony. Before the major competitions the reductions in load, density, volume or frequency of the training in order to achieve optimal performance are made which is called the taper. The aim of taper training is to reduce fatigue and increase physiological adaptation and performance in athletes through intensive training. Since each sport branch has different physiological demands, taper trainings are applied differently in individual and team sports. The effects of these practices may vary in athletes in different branches. In the literature studies, some increases were found in the blood volume and red blood cells values, muscular glycogen deposits, some enzymes, blood lactate and VO2 max. values and the movement economies of athletes. However, in some studies, some decreases were found in the levels of the respiratory threshold, creatine kinase in the blood and the values of the submaximal ventilation, the diastolic and systolic blood pressures of the athletes. Keywords: Taper training, athlete, performance improvement, physiological changes

Effect of dietary lipid levels on growth performance, body composition, and feed utilization of juvenile spotted knifejaw Oplegnathus punctatus

An 8-week feeding trial was carried out with various lipid level (2.30 %, 6.31 %, 8.98 %, 12.93 %, 16.08 %, and 18.99 %) diets to investigate the effects on growth performance, body composition, and feed utilization of juvenile spotted knifejaw Oplegnathus punctatus. After the feeding experiment, the weight gain (WG) and specific growth rate (SGR) significantly elevated with raising the dietary lipid level to 12.93 %, and then decreased when dietary lipid further increased (p < 0.05). The tendencies of feed efficiency (FE), protein efficiency ratio (PER), nitrogen retention (NR), and energy retention (ER) were consistent with the growth performance (p < 0.05). The lipid retention (LR) significantly decreased with dietary lipid increasing (p < 0.05). The fish consuming 2.30 % and 6.31 % lipid diets had significantly elevated hepatosomatic index (HSI) (p < 0.05). The intraperitoneal fat ratio (IPF) increased with the increase of dietary lipid levels (p < 0.05). The elevation of dietary lipid levels significantly increased the lipid concentrations in the whole body, dorsal muscle, and liver (p < 0.05). The highest liver glycogen deposition was found in the 2.30 % dietary lipid treatment (p < 0.05). The fish fed the diets added with 12.93 % lipid diet had the highest serum protein concentration (p < 0.05). The lowest total cholesterol (TC) content in the serum was detected in 2.30 % dietary lipid treatment (p < 0.05). The fish fed diets with 2.30 % or 18.99 % lipid obtained elevated serum AST and ALT activities (p < 0.05). The regression analysis in second order polynomial and two slope broken-line mode based on SGR and FE indicated that diet containing 10.46–12.83 % lipids was optimal for juvenile O. punctatus to obtain best growth performance and feed efficiency.

Abstract 2461: SIRT1 regulates ovarian cancer metastasis via altering the exosome release and glycogen deposition

Abstract Ovarian cancer is the lethal most gynaecological malignancy which exhibits high metastatic rate in peritoneal cavity, especially leading to metastasis including omentum. According to the histopathological segregation of ovarian cancer, epithelial ovarian cancer (EOC) is considered as the most metastatic in both pre- or post-menopausal state, thus several changes in hormonal level such as estrogen, progesterone etc., increase the risk of developing ovarian cancer in women. Obesity or fat deposition are observed to be the after effects of hormonal transition mainly due the depletion of estrogen level. In this context, stored glycogen or deposited fat in the cancer cells has recently drawn the research concern. Few studies suggested that hypoxic environment (HIF-1) could regulate the accumulation of glycogen in cancer cells leading the metastatic micro-environment. The level of HIF-1 could coordinate with the SIRT1 status in tumor dormancy via regulating oxidative stress and metabolic alterations that confers the ovarian cancer stem cell like properties. Interestingly, SIRT1 also controls the secretion of pro-tumorigenic exosomes in cancer invasion. Exosomes, small bioactive extracellular vesicles, can migrate expediently through body barriers and promotes the homing of migrating cancer cells. We observed that the overexpressed SIRT1 in EOC patients has increased concentration of secreted exosomes as compared to heathy individuals. Simultaneously, we have shown how SIRT1 level in ovarian cancer cells co-relate with exosome secretion in regulating glycogen deposition of post-menopausal females. Furthermore, we also found that post-menopausal EOC patients have shown an increased expression of lipids and glycogen in the cancer cells. Therefore, we can conclude that ovarian cancer cell derived exosomes might facilitate fat deposition via regulating their oncogenic cargoes to metastasize ovarian cancer. Citation Format: Sraddhya Roy, Ananya Das, Manisha Vernekar, Subhadip Das, Nabanita Chatterjee. SIRT1 regulates ovarian cancer metastasis via altering the exosome release and glycogen deposition [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2461.

Hepatotoxicity of copper sulfide nanoparticles towards hepatocyte spheroids using a novel multi-concave agarose chip method.

Aim: To explore the hepatotoxicity of copper sulfide nanoparticles (CuSNPs) toward hepatocyte spheroids. Materials & methods: Other than thetraditional agarose method to generate hepatocyte spheroids, we developed a multi-concave agarose chip (MCAC) method to investigate changes in hepatocyte viability, morphology, mitochondrial membrane potential, reactive oxygen speciesand hepatobiliary transporter by CuSNPs. Results: The MCAC method allowed a large number of spheroids to be obtained per sample. CuSNPs showed hepatotoxicity in vitro through a decrease in spheroid viability, albumin/urea production and glycogen deposition. CuSNPs also introduced hepatocyte spheroid injury through alteration of mitochondrial membrane potential and reactive oxygen species, that could be reversed by N-acetyl-l-cysteine. CuSNPs significantly decreased the activity of BSEP transporter by downregulating its mRNA and protein levels. Activity of theMRP2 transporter remained unchanged. Conclusion: We observed the hepatotoxicity of CuSNPs in vitro with associated mechanisms in an advanced 3D culture system.

Wdfy3 regulates glycophagy, mitophagy, and synaptic plasticity

Autophagy is essential to cell function, as it enables the recycling of intracellular constituents during starvation and in addition functions as a quality control mechanism by eliminating spent organelles and proteins that could cause cellular damage if not properly removed. Recently, we reported on Wdfy3’s role in mitophagy, a clinically relevant macroautophagic scaffold protein that is linked to intellectual disability, neurodevelopmental delay, and autism spectrum disorder. In this study, we confirm our previous report that Wdfy3 haploinsufficiency in mice results in decreased mitophagy with accumulation of mitochondria with altered morphology, but expanding on that observation, we also note decreased mitochondrial localization at synaptic terminals and decreased synaptic density, which may contribute to altered synaptic plasticity. These changes are accompanied by defective elimination of glycogen particles and a shift to increased glycogen synthesis over glycogenolysis and glycophagy. This imbalance leads to an age-dependent higher incidence of brain glycogen deposits with cerebellar hypoplasia. Our results support and further extend Wdfy3’s role in modulating both brain bioenergetics and synaptic plasticity by including glycogen as a target of macroautophagic degradation.

A very fruitful liver – hepatic glycogenosis in poorly controlled type 1 diabetes mellitus

Hepatic glycogenosis (HG) is an underrecognised hepatic pathology of diabetes with excess glycogen deposition in the hepatocytes as a result of hyperglycaemia and large doses of insulin administration in type 1 diabetes. This case study reports a 25-year-old female with a constellation of nonspecific symptoms including lethargy, tremors, diaphoresis, and generalized abdominal pain. This in on a background of anti-IA2 antibody positive type 1 diabetes with a HbA1c of 8.9%. The patient also had a persistently elevated lactate level in the absence of sepsis or abdominal pathology. Core liver biopsy showed macrovesicular steatosis, hepatocytes with granular cytoplasm and pseudo inclusions, which stained PAS-positive for glycogen. The hepatocytes also showed features of megamitochondria. Focal lobular inflammation was seen associated with spotty necrosis in one focus. HG arises from disruption of the normal balance between glycogenesis and glycogenolysis, leading to trapping of glycogen within the hepatocytes. This leads to hepatomegaly and consequent liver damage which becomes evident with blood release of aminotransferases.

Substance P-induced lung inflammation in mice is mast cell dependent.

Allergic asthma is a common inflammatory lung disease and a major health problem worldwide. Mast cells (MCs) play a key role in the early-stage pathophysiology of allergic asthma. Substance P (SP) functions in neurogenic inflammation by activating MCs, and therefore, it may to participate in the occurrence and development of asthma. We examined the relationship between SP and lung inflammation, and also whether SP can directly trigger asthma. We measured the number of peripheral blood eosinophils, neutrophils and basophils and evaluated the levels of IgE and SP in blood samples of 86 individuals with allergic asthma. Serum IgE and SP levels were also determined in 29 healthy individuals. C57BL/6 mice were subjected to different doses of SP, and bronchoalveolar lavage fluid (BALF) was collected to count the inflammatory cells. Lung tissues were analysed using histopathological methods to evaluate lung peribronchial inflammation, fibrosis and glycogen deposition. Levels of IgE, interleukin (IL)-1, IL-2, IL-4, IL-5, IL-13, IL-17 and IFN-γ were determined in mouse serum. Substance P levels were increased in the serum samples of patients with asthma. SP induced mouse lung peribronchial inflammation, fibrosis and glycogen deposition, with high levels of Th2-related cytokines such as IL-4, IL-5 and IL-13 observed in the BALF. Furthermore, low level of total IgE was noted in the serum, and SP had little effect on MC-deficient kitW-sh/W-sh mice. Substance P levels increased significantly in serum of asthmatic patients and independently associated with the risk of asthma. Furthermore, SP induced Th2 lung inflammation in mice, which was dependent on MCs.

GLYCOGEN ACCUMULATION IN TRICHOMONAS IS DRIVEN BY THE AVAILABILITY OF EXTRACELLULAR GLUCOSE.

The parasitic protist Trichomonas vaginalis is the causative agent of trichomoniasis, a highly prevalent sexually transmitted infection. The organism is known to accumulate substantial deposits of the polysaccharide glycogen, which is believed to serve as a store of carbon and energy that can be tapped during periods of nutrient limitation. Such nutrient limitation is likely to occur when T. vaginalis is transmitted between hosts, implying that glycogen may play an important role in the lifecycle of the parasite. Both T. vaginalis glycogen synthase and glycogen phosphorylase, key enzymes of glycogen synthesis and degradation, respectively, have been cloned and characterized, and neither enzyme is subject to the post-translational controls found in other, well-characterized eukaryotic systems. Thus, it is unclear how glycogen metabolism is regulated in this organism. Here we use a glucose limitation/re-feeding protocol to show that the activities of key enzymes of glycogen synthesis do not increase during re-feeding when glycogen synthesis is stimulated. Rather, a simple model appears to operate with glycogen storage being driven by the extracellular glucose concentration.

Tissue-Specific Warburg Effect in Breast Cancer and Cancer-Associated Adipose Tissue-Relationship between AMPK and Glycolysis.

Simple SummarySpecific metabolic phenotypes of breast cancer result from local interactions such as cancer-adipocyte cross-talk and systemic metabolic influences such as obesity. Here we examined key regulatory enzymes involved in glucose metabolism in breast cancer tissue and cancer-associated adipose tissue of normal-weight and overweight/obese premenopausal women in comparison to benign breast tumor tissue and adipose tissue of weight-matched women. We show a simultaneous increase in 5′-AMP-activated protein kinase (AMPK) protein expression with glucose utilization favoring glycolysis and pentose phosphate pathway in breast cancer tissue. In parallel, we show an increased AMPK protein expression with glucose utilization favoring the pentose phosphate pathway in cancer-associated adipose tissue. Moreover, specific features of cancer tissue glycolysis and glycogen metabolism differ between normal-weight and overweight/obese women. The results suggest context-dependent induction of tissue-specific Warburg effect in breast cancer and cancer-associated adipose tissue.Typical features of the breast malignant phenotype rely on metabolic reprogramming of cancer cells and their interaction with surrounding adipocytes. Obesity is strongly associated with breast cancer mortality, yet the effects of obesity on metabolic reprogramming of cancer and cancer-associated adipose tissue remain largely unknown. Paired biopsies of breast tumor tissue and adipose tissue from premenopausal women were divided according to pathohistological analyses and body mass index on normal-weight and overweight/obese with benign or malignant tumors. We investigated the protein expression of key regulatory enzymes of glycolysis, pentose phosphate pathway (PPP), and glycogen synthesis. Breast cancer tissue showed a simultaneous increase in 5′-AMP-activated protein kinase (AMPK) protein expression with typical features of the Warburg effect, including hexokinase 2 (HK 2) overexpression and its association with mitochondrial voltage-dependent anion-selective channel protein 1, associated with an overexpression of rate-limiting enzymes of glycolysis (phosphofructokinase 1—PFK-1) and pentose phosphate pathway (glucose-6-phosphate dehydrogenase—G6PDH). In parallel, cancer-associated adipose tissue showed increased AMPK protein expression with overexpression of HK 2 and G6PDH in line with increased PPP activity. Moreover, important obesity-associated differences in glucose metabolism were observed in breast cancer tissue showing prominent glycogen deposition and higher glycogen synthase kinase-3 protein expression in normal-weight women and higher PFK-1 and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) protein expression in overweight/obese women. In conclusion, metabolic reprogramming of glycolysis contributes to tissue-specific Warburg effect in breast cancer and cancer-associated adipose tissue.

Aspects of transition cow metabolomics—Part II: Histomorphologic changes in the liver parenchyma throughout the transition period, in cows with different liver metabotypes and effects of a metaphylactic butaphosphan and cyanocobalamin treatment

The aims of this study were to evaluate histopathologic changes during the transition period, describe the histopathological features of the metabotypes identified in Part I (Schären et al., 2021b), and investigate effects of a metaphylactic treatment with butaphosphan and cyanocobalamin (BCC) on the liver parenchyma. Eighty German Holstein cows (mean 305-d production: 10,957 kg, range: 6,480-15,193 kg; mean lactation number: 3.9, range: 2-9) from a commercial dairy farm in Saxony, Germany, were enrolled in a randomized, prospective, triple-blinded study. Two groups received a treatment with BCC (5 or 10 mL/100 kg of body weight 10% butaphosphan and 0.005% cyanocobalamin, Catosal, Bayer Animal Health, n = 20 each) and one group a placebo treatment (NaCl 0.9%, n = 40). Liver biopsy specimens were collected 14 d antepartum (AP) and 7, 28, and 42 d postpartum (PP), routinely processed for histologic examination, and stained with hematoxylin and eosin, Sudan III, periodic acid-Schiff, and picrosirius red stains. The sections were assessed for fat and glycogen content and degenerative, inflammatory, fibrotic, and proliferative changes. The statistical analysis included the effects of the sampling day, the lactation number, the treatment, and the metabotype (A = medium, B = minor, C = large alterations in the liver metabolome profile between AP and PP status). There was mild to moderate fat infiltration in the liver of 37% of cows in the last 2 wk AP, and moderate to severe fat infiltration in 66% of cows in the first days PP. The degree of fat infiltration increased from 2 wk AP until the end of the first week PP, and then decreased until the end of the study period, at which time about 25% of cows had moderate to severe fatty infiltration. Lipidosis was positively correlated with the severity of liver cell degeneration, and negatively correlated with the degree of glycogen deposits. Complete glycogen depletion of hepatocytes was not observed in cows, even in the presence of severe hepatic lipidosis. Moderate to severe lymphocytic hepatitis was seen in 39% of cows throughout the study period, and cows with lactation numbers 5 or greater had perisinusoidal fibrosis more often than younger cows. Severe fibrosis and cirrhosis of the liver did not occur. Metabotype B animals exhibited a higher chance of fatty infiltration, lower glycogen storage, and perisinusoidal fibrosis and for this metabotype positive correlations were calculated between increased fat deposition in the liver and marked glycogen depletion, and increased degenerative, inflammatory, fibrotic, and proliferative changes of hepatic tissue. For the treatment with BCC, no significant effect was observed. In summary, during the transition period, the liver of dairy cows is characterized by fat accumulation and glycogen depletion and histologic signs of hepatitis and hepatocyte degeneration. These histomorphologic changes were accentuated in animals exhibiting little alterations in their liver metabolome profile across the transition period (metabotype B) and support the assumption of a decreased grass silage quality as a causative factor.

Glycogen synthesis from dihydroxyacetone in isolated rat hepatocytes

Glycogenolysis and gluconeogenesis are sensitive to nutritional state and their balances are disrupted in the liver by different pathological states. The regulation of the balance between glucose production and synthesis of glycogen continues to be a matter of investigation because of its implications in diseases and its value for therapeutics. We used the gluconeogenic precursor dihydroxyacetone (DHA) to study glycogen synthesis in isolated rat hepatocytes under gluconeogenic conditions. We used a glycogen phosphorylase (GP) inhibitor, 2-deoxy-2-fluoro-α-D-glucopyranosyl fluoride (F2Glc) to prevented GP from breaking up glycogen into glucose subunits and had a pool of glucose in the medium. Therefore, we evaluated the contribution of DHA as a unique source of carbohydrates on glycogen metabolism. We showed that DHA increased G6P levels that induced both GS activation and its translocation and, thus, an increment of glycogen deposition in a similar way as glucose did.

Ganglionated Plexi Ablation Suppresses Chronic Obstructive Sleep Apnea-Related Atrial Fibrillation by Inhibiting Cardiac Autonomic Hyperactivation.

Background: Previous studies have reported that right pulmonary artery ganglionated plexi (GP) ablation could suppress the onset of atrial fibrillation (AF) associated with obstructive sleep apnea (OSA) within 1 h.Objective: This study aimed to investigate the effect of superior left GP (SLGP) ablation on AF in a chronic OSA canine model.Methods and Results: Fifteen beagles were randomly divided into three groups: control group (CTRL), OSA group (OSA), and OSA + GP ablation group (OSA + GP). All animals were intubated under general anesthesia, and ventilation-apnea events were subsequently repeated 4 h/day and 6 days/week for 12 weeks to establish a chronic OSA model. SLGP were ablated at the end of 8 weeks. SLGP ablation could attenuate the atrial effective refractory period (ERP) reduction and decrease ERP dispersion, the window of vulnerability, and AF inducibility. In addition, chronic OSA leads to left atrial (LA) enlargement, decreased left ventricular (LV) ejection fraction, glycogen deposition, increased necrosis, and myocardial fibrosis. SLGP ablation reduced the LA size and ameliorated LV dysfunction, while myocardial fibrosis could not be reversed. Additionally, SLGP ablation mainly reduced sympathovagal hyperactivity and post-apnea blood pressure and heart rate increases and decreased the expression of neural growth factor (NGF), tyrosine hydroxylase (TH), and choline acetyltransferase (CHAT) in the LA and SLGP. After SLGP ablation, the nucleotide-binding oligomerization domain (NOD)-like receptor signaling pathway, cholesterol metabolism pathway, and ferroptosis pathway were notably downregulated compared with OSA.Conclusions: SLGP ablation suppressed AF in a chronic OSA model by sympathovagal hyperactivity inhibition. However, there were no significant changes in myocardial fibrosis.