Male Liver Research Articles

HDI-STARR-seq: Condition-specific enhancer discovery in mouse liver in vivo.

STARR-seq and other massively-parallel reporter assays are widely used to discover functional enhancers in transfected cell models, which can be confounded by plasmid vector-induced type-I interferon immune responses and lack the multicellular environment and endogenous chromatin state of complex mammalian tissues. Here, we describe HDI-STARR-seq, which combines STARR-seq plasmid library delivery to the liver, by hydrodynamic tail vein injection (HDI), with reporter RNA transcriptional initiation driven by a minimal Albumin promoter, which we show is essential for mouse liver STARR-seq enhancer activity assayed 7 days after HDI. Importantly, little or no vector-induced innate type-I interferon responses were observed. Comparisons of HDI-STARR-seq activity between male and female mouse livers and in livers from males treated with an activating ligand of the transcription factor CAR (Nr1i3) identified many condition-dependent enhancers linked to condition-specific gene expression. Further, thousands of active liver enhancers were identified using a high complexity STARR-seq library comprised of ~50,000 genomic regions released by DNase-I digestion of mouse liver nuclei. When compared to stringently inactive library sequences, the active enhancer sequences identified were highly enriched for liver open chromatin regions with activating histone marks (H3K27ac, H3K4me1, H3K4me3), were significantly closer to gene transcriptional start sites, and were significantly depleted of repressive (H3K27me3, H3K9me3) and transcribed region histone marks (H3K36me3). HDI-STARR-seq offers substantial improvements over current methodologies for large scale, functional profiling of enhancers, including condition-dependent enhancers, in liver tissue in vivo, and can be adapted to characterize enhancer activities in a variety of species and tissues by selecting suitable tissue- and species-specific promoter sequences.

Letter to the Editor: “Effect of grape seed extract on β-catenin gene expression and hyperglycemia in rats induced by streptozotocin”

This letter is related to the article titled "Effect of grape seed extract on β-catenin gene expression and hyperglycemia in rats induced by streptozotocin" published in Volume 39, Is sue 2 of the Eurasian Journal of Veterinary Sciences in 2023. First of all, we would like to thank you for the study whose aim was to determine the effect of grape seed extract (GSE) on body weight, blood glucose, and β-catenin gene expres sion in experimental Type 2 diabetic rats. We read this ar ticle with interest and have one suggestion for the authors. The authors reported that the effect of grape seed extract on blood glucose, body weight, and B-catenin gene expression levels determined in the Wnt/β-catenin signaling pathway in Type 2 Diabetes Mellitus. Instead of examining the expression levels of the tissue ma terials obtained due to the experimental design at a single gene level, next-generation sequencing, and total transcrip tome level will allow the expression levels of many genes expressed in the relevant pathway to be determined. In this regard, a literature search on the Web of Science with the keywords "streptozotocin", "diabetes", and "RNA-seq" reve aled 29 studies in the last four years. Among these studies; there is a study on RNA-seq analysis in diabetic male mouse liver (Ge et al 2020). Again, different functional, cellular, and molecular models of diabetic cardiomyopathy were exami ned in Type I and Type II mouse models administered with Streptozotocin (Marino et al 2023). Elzinga et al. (2023) reported research results in which diabetic kidney disease and neuropathy were examined at the transcriptome level in type I and type II diabetic mice. In the currently planned and realized experimental design, it is recommended that next-generation sequencing analyses in liver and pancre atic tissues be performed in a short time in terms of cost, time, and more information content. It is envisaged that further investigation of Type 2 Diabetes Mellitus disease at the transcriptome level in animal experimental models with grape seed extract and publication of the analysis results in international journals will have a serious interest potential for researchers working on this subject

Detraining after short-term exercise induces hyperphagia and obesity with fatty liver and brown adipose tissue whitening in young male OLETF rats.

This study examined the effects of exercise and detraining at a young age on fat accumulation in various organs. Four-week-old male Otsuka Long-Evans Tokushima Fatty (OLETF) rats were assigned to either the non-exercise sedentary (OLETF Sed) or exercise groups. The exercise group was subdivided into two groups: exercise between 4 and 12 weeks of age (OLETF Ex) and exercise between 4 and 6 weeks of age followed by non-exercise between 6 and 12 weeks of age (OLETF DT). Body weight was significantly lower in the OLETF Ex group than in the OLETF Sed group at 12 weeks of age. Fat accumulation in the epididymal white adipose tissue, liver, and brown adipose tissue was suppressed in the OLETF Ex group. During the exercise period, body weight and food intake in the OLETF DT group were significantly lower than those in the OLETF Sed group. However, food intake was significantly higher in the OLETF DT group than in the OLETF Sed group after exercise cessation, resulting in extreme obesity with fatty liver and brown adipose tissue whitening. Detraining after early-onset exercise promotes hyperphagia, causing extreme obesity. Overeating should be avoided during detraining periods in cases of exercise cessation at a young age.

Methanolic leaf extract of Vernonia amygdalina mitigates cadmium-induced oxidative stress in the liver and kidney of adult male Wistar rats

ABSTRACT Cadmium is a hazardous metal swiftly generated through industrialization and persists in the environment due to its non-biodegradability. Cadmium induces liver and kidney damage primarily by triggering free radical production causing oxidative stress. This study explored the potential of methanolic extract of Vernonia amygdalina in ameliorating cadmium-induced oxidative stress. Twenty adult male Wistar rats were used for this experiment, with each group consisting of five rats. The first group was the unexposed control group given standard rat chow and distilled water ad libitum while the test groups (II-IV) were orally administered with cadmium at standard doses of 12 mg/kg/BW. Vernonia amygdalina at standard doses of 100 mg/kg/BW and 200 mg/kg/BW was administered to rats in groups III and IV respectively. After 28 days, the rats were euthanized using cervical dislocation and the livers and kidneys were analyzed for histological and mRNA analysis of tumor necrosis factor-alpha (TNF-α) and interleukin-10 (IL-10). The results showed significant pathological changes in the histoarchitecture of the livers and kidneys, indicating oxidative stress and the effectiveness of Vernonia amygdalina, which mitigated cadmium-induced oxidative damage by modulating TNF-α and IL-10 expression and ameliorating the histological alteration seen in the studied organs.

Integrated liver and serum proteomics uncover sexual dimorphism and alteration of several immune response proteins in an aging Werner syndrome mouse model.

Werner syndrome (WS) is a progeroid disorder caused by mutations in a protein containing both a DNA exonuclease and DNA helicase domains. Previous studies indicated that males lacking the helicase domain of the Wrn protein orthologue exhibited hepatic transcriptomic and metabolic alterations. In this study, we used a label-free liquid chromatography-tandem mass spectrometry approach to uncover proteins abundance associated with specific biological processes that differed depending on the age (four or ten months) and/or the genotype (wild type or Wrn mutant) in the serum and liver of mice. Principal component analysis of the proteomic data from both serum and hepatic tissue revealed a sexual dimorphism regardless of the age and the genotype of the mice. Moreover, although all Wrn mutant mice exhibited fatty liver by the age of ten months, a significant age and genotype dependent enrichment of proteins involved in lipid and fatty acid metabolic processes were uncovered only in males. Also, a genotype dependent increase in serum oxidant detoxification processes was observed in the serum of Wrn mutant males. Despite these sexual differences, several aspects of the immune system were affected in both females and males. Finally, an increase of specific immunoglobulin molecules was common in the liver and serum of both older Wrn mutant females and males. Such results suggest that specific immunoglobulin variants maybe associated with fatty liver progression in WS.

Changes in CpG Methylation of the Vitellogenin 1 Promoter in Adult Male Zebrafish after Exposure to 17α-Ethynylestradiol.

Numerous pharmaceutical and industrial chemicals are classified as endocrine-disrupting chemicals (EDCs) that interfere with hormonal homeostasis, leading to developmental disorders and other pathologies. The synthetic estrogen 17α-ethynylestradiol (EE2) is used in oral contraceptives and other hormone therapies. EE2 and other estrogens are inadvertently introduced into aquatic environments through municipal wastewater and agricultural effluents. Exposure of male fish to estrogens increases expression of the egg yolk precursor protein vitellogenin (Vtg), which is used as a molecular marker of exposure to estrogenic EDCs. The mechanisms behind Vtg induction are not fully known, and we hypothesized that it is regulated via DNA methylation. Adult zebrafish were exposed to either dimethyl sulfoxide or 20 ng/L EE2 for 14 days. Messenger RNA (mRNA) expression and DNA methylation were assessed in male zebrafish livers at 0, 0.25, 0.5, 1, 4, 7, and 14 days of exposure; and those of females were assessed at 13 days (n ≥ 4/group/time point). To test the persistence of any changes, we included a recovery group that received EE2 for 7 days and did not receive any for the following 7 days, in the total 14-day study. Methylation of DNA at the vtg1 promoter was assessed with targeted gene bisulfite sequencing in livers of adult male and female zebrafish. A significant increase in vtg1 mRNA was observed in the EE2-exposed male fish as early as 6 h. Interestingly, DNA methylation changes were observed at 4 days. Decreases in the overall methylation of the vtg1 promoter in exposed males resulted in levels comparable to those in female controls, suggesting feminization. Importantly, DNA methylation levels in males remained significantly impacted after 7 days post-EE2 removal, unlike mRNA levels. These data identify an epigenetic mark of feminization that may serve as an indicator of not only estrogenic exposure but also previous exposure to EE2. Environ Toxicol Chem 2024;43:1547-1556. © 2024 SETAC. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA.

Effect of Mesenchymal Stem Cells-Conditioned Medium versus Silymarin on Experimentally Induced Fatty Liver in Adult Male Albino Rats (A Histological and Immunohistochemical Study)

Effect of Mesenchymal Stem Cells-Conditioned Medium versus Silymarin on Experimentally Induced Fatty Liver in Adult Male Albino Rats (A Histological and Immunohistochemical Study)

Evaluation of propolis hepatotoxicity in male rats

Background: Propolis is a natural resinous combination that honeybees produce from the materials they gather from plant parts, buds, and exudates. Numerous favorable pharmacological qualities have been demonstrated for propolis. Objectives: This study aimed to investigate the hepatotoxic effects of ethanolic propolis extract on the liver of male rats. Methods: In this research, 40 male rats were divided randomly into five groups: 1. Control 2. Sham (solvent), and 3. Three experimental groups (ethanolic propolis extract at doses of 50,100 and 200 mg/kg). All materials were administered by oral gavage once daily for 13 consecutive days. On the 14th day, blood sampling was performed to measure serum levels of liver function enzymes and triglycerides. After deep induction of anesthesia, the liver of the rats was removed for histopathological studies. Data were analyzed using ANOVA and Tukey’s test (p <0.05). Results: The data showed that the administration of propolis significantly increased the serum levels of aminotransferases in a dose-dependent manner and decreased triglycerides, accompanied by pathological changes. Conclusions: The results of this study indicate that the administration of propolis is associated with liver toxicity, and it seems that it should be consumed more carefully. Keywords: Alanine aminotransferase, Aspartate aminotransferase, Propolis, Hepatotoxicity Triglycerides

Determination of Flavonoids and Phenolic Acids in the Liver of Wistar Rats after a Dietary Enrichment with Corinthian Currant (Vitis vinifera L., var. Apyrena): A Liquid Chromatography-Tandem Mass Spectrometry Study.

Corinthian currants are dried fruits produced from Vitis vinifera L. var. Apyrena grape. This study investigated the distribution of phenolic compounds in male Wistar rat livers following two distinct Corinthian currant long-term dietary intake protocols (3 and 10% w/w). Method optimization, comparing fresh and lyophilized tissues, achieved satisfactory recoveries (>70%) for most analytes. Enzymatic hydrolysis conditions (37 °C, pH 5.0) minimally affected phenolics, but enzyme addition showed diverse effects. Hydrolyzed lyophilized liver tissue from rats consuming Corinthian currants (3 and 10% w/w) exhibited elevated levels of isorhamnetin (20.62 ± 2.27 ng/g tissue and 33.80 ± 1.38 ng/g tissue, respectively), along with similar effects for kaempferol, quercetin, and chrysin after prolonged Corinthian currant intake. This suggests their presence as phase II metabolites in the fasting-state liver. This study is the first to explore phenolic accumulation in rat liver, simulating real conditions of dried fruit consumption, as seen herein with Corinthian currant.

The state of cancer care in Russia: liver cancer - (From 22). Age-related features of the observed and relative one-year and five-year survival of patients. (clinical and population-based study). Part 2.

Survival rate calculations for liver cancer patients at the federal level in the Russian Federation are only conducted in the North-Western Federal District and a few other individual administrative territories. In the first part of our study, we presented a detailed analysis of the prevalence of liver cancer, noting that the levels of standardized incidence and mortality rates in Russia and the North-Western Federal District are virtually identical, allowing us to consider the results of our study as nationally representative. The conducted research confirmed the seriousness of the issue of liver cancer among the population. However, certain advancements in patient treatment were recorded, with a significant increase in survival rates, primarily for one-year survival. For example, in the North-Western Federal District, one-year survival increased from 13.6% in 2000 to 23.1% in 2022, representing a 69.8% increase. At the same time, five-year survival rates remained largely unchanged. Increases in survival rates were observed in most age groups. A significant deficiency in the initial patient record-keeping was identified - insufficiently accurate determination of the stage of liver cancer. One-year survival rates for male and female liver cancer patients (22.7% and 22.1%) were nearly equal, as were the survival rates for liver cancer patients residing in urban and rural areas. Therefore, further improvement in the outcomes of treatment for this complex patient group can only be achieved through the implementation of regional and federal screening and epidemiological surveillance programs among populations at high and extremely high risk of developing hepatocellular carcinoma.

EVALUATION OF ACUTE AND SUB-ACUTE TOXICITY OF METHANOLIC LEAF EXTRACT OF BAMBUSA VULGARIS ON MALE WISTAR RATS

Objectives: The study aims to evaluate the potential toxicity of methanolic extract of leaves of Bambusa vulgaris in male Wistar rats. Methods: The methanolic extract was prepared for acute and sub-acute toxicity to determine the lethal dose. During the acute toxicity study, male rats were orally administered with leaf extracts at single doses of 100, 500,800, and 2000 mg/kg according to OECD Guidelines 425 for 14 days. Sub-acute toxicity dose at 200, 400, and 800 mg/kg was studied by daily dosing on male rats for 30 days. Results: The acute toxicity study revealed no lethal effects and behavioral signs of toxicity at the tested doses indicating that LD50 is <2000 mg/kg. In a sub-acute study, non-significant changes in the body weight (p<0.05), body weight gain percentage (p<0.05) food intake (p<0.05), and relative organ weight percentage (p<0.05), and hematological parameters (p<0.05) of the rats were observed. A significant (p<0.05) increase in ALT level was observed dose at 800 mg/kg. Histopathological analysis showed mild cell distortion in the liver and kidney in male rats treated at 800 mg/kg of extract. Conclusion: Our study suggests that oral administration of methanolic leaf extract Bambusa vulgais is safe. However, a dose of 800 mg/kg may be toxic for the liver and kidney.

Tissue-specific differences in Ca2+ sensitivity of the mitochondrial permeability transition pore (PTP). Experiments in male rat liver and heart.

Permeability transition pore (PTP) opening dissipates ion and electron gradients across the internal mitochondrial membrane (IMM), including excess Ca2+ in the mitochondrial matrix. After opening, immediate PTP closure must follow to prevent outer membrane disruption, loss of cytochrome c, and eventual apoptosis. Flickering, defined as the rapid alternative opening/closing of PTP, has been reported in heart, which undergoes frequent, large variations in Ca2+. In contrast, in tissues that undergo depolarization events less often, such as the liver, PTP would not need to be as dynamic and thus these tissues would not be as resistant to stress. To evaluate this idea, it was decided to follow the reversibility of the permeability transition (PT) in isolated murine mitochondria from two different tissues: the very dynamic heart, and the liver, which suffers depolarizations less frequently. It was observed that in heart mitochondria PT remained reversible for longer periods and at higher Ca2+ loads than in liver mitochondria. In all cases, Ca2+ uptake was inhibited by ruthenium red and PT was delayed by Cyclosporine A. Characterization of this phenomenon included measuring the rate of oxygen consumption, organelle swelling and Ca2+ uptake and retention. Results strongly suggest that there are tissue-specific differences in PTP physiology, as it resists many more Ca2+ additions before opening in a highly active organ such as the heart than in an organ that seldom suffers Ca2+ loading, such as the liver.

Signaling of GPR17 in Hepatic and Renal Tissues

G protein-coupled receptors (GPCRs) are the largest class of proteins in the human body and are considered to be excellent targets for drug discovery. However, the vast majority of GPCRs are understudied. Using a large-scale TaqMan array screen on both male and female C57BL6 murine livers, we identified GPR17 as one of the most highly expressed orphan receptors within the liver (ΔCt = 19.9, normalized to GAPDH). GPR17 expression in the liver was confirmed using reverse transcriptase polymerase chain reaction (RT-PCR) and was also identified in murine whole kidney from both males and females indicating that GPR17 RNA expression is not sex-dependent. While GPR17 has known roles in oligodendrocyte maturation and myelination in the central nervous system, its functions within the liver and kidney are undetermined. To investigate its physiological functions, we started by uncovering its localization within the liver and kidney and identifying its ligand profile. A series of RNAScope in-situ hybridization assays were performed to localize GPR17 within hepatic tissue. Gpr17 mRNA was found within hepatocytes indicating that its highest expression is in the parenchymal cells of the liver. To identify possible activators for GPR17, we cloned the coding region into both flag-tagged PME18s and the Presto-Tango vectors. Live-cell surface labeling of HEK293T expressing the GPR17 constructs confirmed robust surface expression on the plasma membrane facilitating ligand screening. GPR17 downstream signaling has yet to be studied. Thus, we used the PRESTO-TANGO assay, an exogenous chemical ligand screening assay that relies on a luminescence readout to indicate -arrestin recruitment and receptor activation. A large, non-biased chemical screen of over 500 physiological compounds was tested for activation of GPR17. To date, we identified seven exogenous activators of GPR17: 3-[methyl(1,2,3,4- tetrahydronaphthalen-2-yl)amino]pyrazine-2-carbonitrile, 4-(4-fluorophenyl)-5-methyl-N-(2-methyl-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)thiophene-2-carboxamide,({1-[(2-chlorophenyl)methyl]-3,5-dimethyl-1H-pyrazol-4-yl}methyl)(methyl)[(1-methyl-1H-imidazol-2-yl)methyl]amine,N,N-dimethyl-6-[(5-phenyl-1,3-oxazol-2-yl)methyl]-5H,6H,7H,8H-pyrido[4,3-d]pyrimidin-4-amine,4-phenyl-1-[6-(pyrrolidin-1-yl)-9H-purin-9-yl]butan-2-ol, 2-(3-fluorophenyl)-3-methyl-1-{[1,2,4]triazolo[4,3-a]pyridine-6-carbonyl}pyrrolidine, and 2,6,6-trimethyl-N-({2-methyl-4H,5H,6H,7H-pyrazolo[1,5-a]pyrimidin-6-yl}methyl)-4,5,6,7-tetrahydro-1-benzofuran-4-amine. Ongoing studies are aimed at narrowing down the in vivo activators of GPR17 to ultimately shed light on the physiological functions that this receptor plays in the kidney and liver. NIH/NCATS. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Phenotypic Sexual Dimorphism in Response to High Fat Diet in C57BL/6J Mice Lacking Liver Androgen Receptor

Previous research has indicated that liver androgen receptors may play a role in modulating disease. This study aims to investigate the pathophysiology of high fat diet (HFD) induced dysglycemia in male and female liver androgen receptor knockout (LivARKO) mice. We performed metabolic tests on LivARKO female and male mice fed a high fat diet (HFD) or a control diet (CD) (from Research Diets Inc) during months 1 or 2 after starting the diet. Additionally, we performed western blot and qRT-PCR analysis on the livers of the mice to examine intermediates in the insulin signaling pathway. LivARKO-HFD female mice displayed no difference in glucose tolerance compared to female LivARKO-Control mice. Whereas in WT female mice, HFD impaired glucose tolerance (IGT). Our data suggests that starting at one-month LivARKO may be protecting female mice from HFD-induced metabolic dysfunction. LivARKO-HFD female mice displayed significantly worse insulin sensitivity at 15 minutes compared to LivARKO-Control (Con) female mice; but, strangely, LivARKO-HFD female mice had significantly better insulin sensitivity at 60 and 90 minutes compared to LivARKO-Con female mice. Despite protecting against IGT, LivARKO did not protect against HFD-induced hyperinsulinemia (HI) in female mice. In contrast to females, male LivARKO-HFD mice displayed impaired glucose tolerance (IGT) compared to male LivARKO-Con mice. Thus, LivARKO is not protective against HFD-induced glucose metabolic dysfunction in male mice. Lastly, LivARKO-HFD female mice maintained hepatic insulin sensitivity whereas LivARKO-HFD male mice displayed hepatic insulin resistance. These findings suggest that LivARKO delayed the onset of HFD-Induced dysglycemia in female mice. Funding: S.A. grant support from the National Science Foundation (NSF) Inclusion across the Nation of Communities of Learners of Underrepresented Discoverers in Engineering and Science (INCLUDES, HRD-1931045). S.A. grant from the National Institutes of Health (NIH) 1R01DK126892-01A1. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

ADGRA3: In vivo hepatic localization, functional importance, and in vitro activation

G protein-coupled receptors (GPCRs), which encode for more than 3% of all human genes, are the largest protein family in the human genome. However, most GPCRs remain under-characterized with little known about their expression, agonists, and physiological function. To identify understudied receptors in the liver, the largest metabolic organ in the body, we performed a TaqMan array screen of more than 300 GPCRs in both male and female C57BL6 murine livers. Among the most highly expressed orphan receptors is ADGRA3 (GPR125), an adhesion GPCR (aGPCR), whose expression is comparable to that of the glucagon receptor, with ΔCt values of 17.2 and 16.0 respectively. aGPCRs possess a unique N-terminal domain that includes a GPCR auto proteolysis-inducing (GAIN) domain, which contains a GPS cleavage site that allows for the cleavage and non-covalent attachment of the N-Terminal (NTF) and C-Terminal Fragments. It is theorized that the displacement of the NTF exposes a tethered ligand, which ultimately results in ADGRA3 activation. ADGRA3 has an unconventional GPS site and it was unclear if this receptor follows a similar activation sequence. To test this, we synthesized peptides corresponding to the β-strand immediately following the predicted GPS domain and tested for activation using the PRESTO-TANGO assay, a luminescence readout to indicate receptor activation. We confirmed that ADGRA3 is activated by a tethered ligand, observing maximal activation with the complete β-strand and 2 additional C-Terminal amino acids. We then further employed the PRESTO-TANGO assay to deorphanize ADGRA3 by identifying non-self activators in a large, unbiased chemical screen (>500 chemicals). This resulted in the identification of more than 25 unique ligands whose selective binding results in ADGRA3 activation. To date, our studies show that ADGRA3 is best activated by C16H20N2O3. By using RNA scope, we localized Adgra3 transcripts to the cuboidal endothelial cells in the lining of the bile duct and hepatocytes. This localization was confirmed through antibody staining, which revealed that ADGRA3 localizes to the basolateral membrane of cholangiocytes. To investigate physiological function of hepatic ADGRA3, we are characterizing the C57BL6-ADGRA3-GreenLantern mouse (GL-KO), where insertion of a GreenLantern tag within ADGRA3 is predicted to disrupt functionality. To visualize the impacts of ADGRA3 disruption, we employed functional Magnetic Resonance Imaging (MRI) to track the uptake of gadoxetate disodium (EOVIST). Injection of exogenous EOVIST is selectively taken up by the liver through organic anion transporting polypeptides (OATP) and is further cleared into the gallbladder. In wildtype ADGRA3 mice, EOVIST was rapidly taken up by the liver and was found within the common bile duct entering the gallbladder by 60 minutes. In the GL-KO, this clearance was delayed with minimal accumulation in the common bile duct. Given the localization of ADGRA3 within the cholangiocytes, this raises the possibility that ADGRA3 may play an essential role in hepatic development and biliary repair mechanisms. NIH/NCATS. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Pneumocystis jirovecii Pneumonia in a Liver Transplant Recipient With an Adverse Reaction to Trimethoprim/Sulfamethoxazole Treated With a Sulfonamide Desensitization Protocol: Case Report

Pneumocystis jirovecii pneumonia (PJP) is an opportunistic fungal infection that, in immunocompromised patients, can progress to respiratory failure and death. Since trimethoprim/sulfamethoxazole (TMP/SMX) chemoprophylaxis has become a standard management, the prognosis has improved. However, there are patients with a history of TMP/SMX intolerance who cannot receive chemoprophylaxis. We report on a 53-year-old male liver recipient treated with a standard triple immunosuppressive regimen in whom TMP/SMX was waived because of a history of allergy manifested as a generalized rash with edema more than 30 years ago. At transplantation, the immunologic risk was assessed as low, and liver graft function was normal. In the third month after engraftment, he developed dyspnea at rest required constant passive oxygen therapy. Ceftriaxone, azithromycin, and clindamycin were implemented. Mycophenolate acid was stopped, and tacrolimus was reduced. High-resolution computed tomography revealed interstitial pneumonia. Pneumocystis jirovecii pneumoniae was diagnosed from bronchoalveolar lavage. Instead of TMP/SMX, pentamidine and caspofungin were also used for PJP, with no improvement. After 3 weeks, the patient deteriorated. Because of his life-threatening condition, TMP/SMX was introduced in the sulfonamide desensitization protocol, including hydrocortisone and clemastinum. Within 4 days, the patient stabilized with no signs of TMP/SMX intolerance. Pneumonia subsided within a month, and TMP/SMX was prescribed lifelong. Prophylaxis for PJP with TMP/SMX still remains an important issue in transplant recipients. Adverse reaction to TMP/SMX in the past is not always a contraindication to reintroducing prophylaxis. The decision of prophylaxis avoidance should be analyzed carefully; in uncertain cases, a sulfonamide desensitization protocol should be considered.

The Association of Cardiometabolic, Diet and Lifestyle Parameters With Plasma Glucagon-like Peptide-1: An IMI DIRECT Study.

The role of glucagon-like peptide-1 (GLP-1) in type 2 diabetes (T2D) and obesity is not fully understood. We investigate the association of cardiometabolic, diet, and lifestyle parameters on fasting and postprandial GLP-1 in people at risk of, or living with, T2D. We analyzed cross-sectional data from the two Innovative Medicines Initiative (IMI) Diabetes Research on Patient Stratification (DIRECT) cohorts, cohort 1 (n = 2127) individuals at risk of diabetes; cohort 2 (n = 789) individuals with new-onset T2D. Our multiple regression analysis reveals that fasting total GLP-1 is associated with an insulin-resistant phenotype and observe a strong independent relationship with male sex, increased adiposity, and liver fat, particularly in the prediabetes population. In contrast, we showed that incremental GLP-1 decreases with worsening glycemia, higher adiposity, liver fat, male sex, and reduced insulin sensitivity in the prediabetes cohort. Higher fasting total GLP-1 was associated with a low intake of wholegrain, fruit, and vegetables in people with prediabetes, and with a high intake of red meat and alcohol in people with diabetes. These studies provide novel insights into the association between fasting and incremental GLP-1, metabolic traits of diabetes and obesity, and dietary intake, and raise intriguing questions regarding the relevance of fasting GLP-1 in the pathophysiology T2D.

Induction of the Inflammasome Pathway by Tyrosine Kinase Inhibitors Provides an Actionable Therapeutic Target for Hepatocellular Carcinoma.

During the last decade, tyrosine kinase inhibitors (TKIs) sorafenib and regorafenib have been standard systemic treatments for advanced hepatocellular carcinoma (HCC). Previous data associated sorafenib with inflammasome activation. However, the role of the inflammasome in sorafenib and regorafenib signaling has not been described in liver cancer patients. For this purpose, we analyzed inflammasome-related transcriptomic changes in a murine HCC model. Our data confirmed inflammasome activation after both TKI treatments, sharing a similar pattern of increased gene expression. According to human database results, transcriptional increase of inflammasome genes is associated with poorer prognosis for male liver cancer patients, suggesting a sex-dependent role for inflammasome activation in HCC therapy. In biopsies of HCC and its surrounding tissue, we detected durable increases in the inflammasome activation pattern after sorafenib or regorafenib treatment in male patients. Further supporting its involvement in sorafenib action, inflammasome inhibition (MCC950) enhanced sorafenib anticancer activity in experimental HCC models, while no direct in vitro effect was observed in HCC cell lines. Moreover, activated human THP-1 macrophages released IL-1β after sorafenib administration, while 3D Hep3B spheres displayed increased tumor growth after IL-1β addition, pointing to the liver microenvironment as a key player in inflammasome action. In summary, our results unveil the inflammasome pathway as an actionable target in sorafenib or regorafenib therapy and associate an inflammasome signature in HCC and surrounding tissue with TKI administration. Therefore, targeting inflammasome activation, principally in male patients, could help to overcome sorafenib or regorafenib resistance and enhance the efficacy of TKI treatments in HCC.

Spontaneously hypertensive rats exhibit increased liver flavin monooxygenase expression and elevated plasma TMAO levels compared to normotensive and Ang II-dependent hypertensive rats.

Background: Flavin monooxygenases (FMOs) are enzymes responsible for the oxidation of a broad spectrum of exogenous and endogenous amines. There is increasing evidence that trimethylamine (TMA), a compound produced by gut bacteria and also recognized as an industrial pollutant, contributes to cardiovascular diseases. FMOs convert TMA into trimethylamine oxide (TMAO), which is an emerging marker of cardiovascular risk. This study hypothesized that blood pressure phenotypes in rats might be associated with variations in the expression of FMOs. Methods: The expression of FMO1, FMO3, and FMO5 was evaluated in the kidneys, liver, lungs, small intestine, and large intestine of normotensive male Wistar-Kyoto rats (WKY) and two distinct hypertensive rat models: spontaneously hypertensive rats (SHRs) and WKY rats with angiotensin II-induced hypertension (WKY-ANG). Plasma concentrations of TMA and TMAO were measured at baseline and after intravenous administration of TMA using liquid chromatography-mass spectrometry (LC-MS). Results: We found that the expression of FMOs in WKY, SHR, and WKY-ANG rats was in the descending order of FMO3 > FMO1 >> FMO5. The highest expression of FMOs was observed in the liver. Notably, SHRs exhibited a significantly elevated expression of FMO3 in the liver compared to WKY and WKY-ANG rats. Additionally, the plasma TMAO/TMA ratio was significantly higher in SHRs than in WKY rats. Conclusion: SHRs demonstrate enhanced expression of FMO3 and a higher plasma TMAO/TMA ratio. The variability in the expression of FMOs and the metabolism of amines might contribute to the hypertensive phenotype observed in SHRs.

Superoxidase dismutases (SODs) in the European eel: Gene characterization, expression response to temperature combined with hormonal maturation and possible migratory implications

Superoxide dismutases (SODs) are antioxidant enzymes that protect cells from oxidation. Three SODs have been identified in mammals, but there is limited information in teleosts. This study investigates SODs in the European eel and their expression patterns during testis maturation. Phylogenetic and synteny analyses revealed SODs paralogs and their evolution in vertebrates. The eel possesses one SOD1 and two SOD2/3 (a and b), indicating SOD2 and SOD3 duplication in elopomorphs. SODs expression were then evaluated in various male and female tissues. SOD1 is more expressed in females, while SOD2a and SOD2b dominate brain-pituitary-gonad tissues in both sexes. SOD3a showed predominant expression in the ovary and the male livers, whereas SOD3b was found in the pituitary and brain of both sexes. The effects of different maturation protocols (standard hormonal treatment vs. same protocol preceded with cold seawater pre-treatment) on SODs expression during testis maturation were evaluated. Salinity increase at the onset of standard treatment at 20 °C, simulating early migration, upregulated SOD1, SOD2a, and SOD2b, coinciding with spermatogonia type A differentiated cells dominance. Thereafter, SOD2a and SOD3a decreased, while SOD2b increased during hormonal treatment-induced spermatogenesis. Pre-treatment with seawater at 10 °C, mimicking the conditions at the beginning of the seawater migration, downregulated SOD1 but increased SOD3a expression. Finally, the standard hormonal treatment, replicating spawning at higher temperatures, downregulated SOD1 in eels without any pre-treatment while SOD2a expression increased in pre-treated eels. This study revealed tissue-specific, sex-dependent, and maturation-related SOD expression patterns, predicting SODs dynamic expression profiles during their reproductive migration.