Different Tissues Of Mice Research Articles

Mathematical modeling of endogenous and exogenously administered T cell recirculation in mouse and its application to pharmacokinetic studies of cell therapies

IntroductionIn vivo T cell migration has been of interest to scientists for the past 60 years. T cell kinetics are important in the understanding of the immune response to infectious agents. More recently, adoptive T cell therapies have proven to be a most promising approach to treating a wide range of diseases, including autoimmune and cancer diseases, whereby the characterization of cellular kinetics represents an important step towards the prediction of therapeutic efficacy. MethodsHere, we developed a physiologically-based pharmacokinetic (PBPK) model that describes endogenous T cell homeostasis and the kinetics of exogenously administered T cells in mouse. Parameter calibration was performed using a nonlinear fixed-effects modeling approach based on published data on T cell kinetics and steady-state levels in different tissues of mice. The Partial Rank Correlation Coefficient (PRCC) method was used to perform a global sensitivity assessment. To estimate the impact of kinetic parameters on exogenously administered T cell dynamics, a local sensitivity analysis was conducted. ResultsWe simulated the model to analyze cellular kinetics following various T cell doses and frequencies of CCR7+ T cells in the population of infused lymphocytes. The model predicted the effects of T cell numbers and of population composition of infused T cells on the resultant concentration of T cells in various organs. For example, a higher percentage of CCR7+ T cells among exogenously administered T lymphocytes led to an augmented accumulation of T cells in the spleen. The model predicted a linear dependence of T cell dynamics on the dose of adoptively transferred T cells. DiscussionThe mathematical model of T cell migration presented here can be integrated into a multi-scale model of the immune system and be used in a preclinical setting for predicting the distribution of genetically modified T lymphocytes in various organs, following adoptive T cell therapies.

Comparative evaluation of silver nanoparticles and human platelet rich-plasma versus traditional therapy in the treatment of murine chronic toxoplasmosis.

Toxoplasmosis is a worldwide parasitic disease infecting about one-third of the human population. At present, licensed medications are incapable of curing human chronic infection. The present work aimed to evaluate for the first time the combination between (spiramycin and human platelet rich plasma), in addition to (spiramycin and silver-nanoparticles) in treating murine experimental toxoplasmosis using parasitological, biochemical, histopathological and immunohistochemical studies. Seventy-seven Swiss albino male mice divided into seven groups according to the treatment used as follows: (GI): control negative; (GII): control infected; (GIII): spiramycin; (GIV): Silver nanoparticles (AgNPs); (GV): Human platelet-rich plasma (HPRP); (GVI): combined spiramycin and AgNPs; (GVII): combined spiramycin and HPRP. Obtained results demonstrated that (spiramycin and AgNPs) treated group showed significant reduction of T. gondii tissue cysts number, the lowest level of serum malondialdehyde, remarkable improvement in pathological changes in different tissues of mice e.g. brain and liver and weak expression of EGFR in brain tissues of mice compared to control infected group. Moreover, AgNPs administered alone produced minimal anti-Toxoplasma results, whereas their combination with spiramycin exhibited significant therapeutic efficacy. In conclusion, combination therapy of spiramycin and AgNPs may represent a unique possible adjuvant therapy for reducing the pathogenic, toxic, and inflammatory consequences of toxoplasmosis on the brain and liver tissues in immunocompetent mice, and the expression of EGFR in brain tissues of mice is a good tool for evaluating the therapeutic improvement of murine toxoplasmosis.

Chronic sleep deprivation induces alterations in DNA and RNA modifications by liquid chromatography-mass spectrometry analysis

Sleep deprivation (SD) is a widespread issue that disrupts the lives of millions of people. These effects initiate as changes within neurons, specifically at the DNA and RNA level, leading to disruptions in neuronal plasticity and the dysregulation of various cognitive functions, such as learning and memory. Nucleic acid epigenetic modifications that could regulate gene expression have been reported to play crucial roles in this process. However, there is a lack of comprehensive research on the correlation of SD with nucleic acid epigenetic modifications. In the current study, we aimed to systematically investigate the landscape of modifications in DNA as well as in small RNA molecules across multiple tissues, including the heart, liver, kidney, lung, hippocampus, and spleen, in response to chronic sleep deprivation (CSD). Using liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis, we characterized the dynamic changes in DNA and RNA modification profiles in different tissues of mice under CSD stress. Specifically, we observed a significant decrease in the level of 5-methylcytosine (5mC) and a significant increase in the level of 5-hydroxymethylcytosine (5hmC) in the kidney in CSD group. Regarding RNA modifications, we observed an overall increased trend for most of these significantly changed modifications across six tissues in CSD group. Our study sheds light on the significance of DNA and RNA modifications as crucial epigenetic markers in the context of CSD-induced stress.

Investigating the protective effects ofElaeagnus angustifoliafruit extract on hematological parameters and damage of different tissues of male mice exposed to graphene oxide nanoparticles

AbstractThe aim of this study is to investigate the protective effects ofElaeagnus angustifoliafruit extract (EAFE) on hematology, biochemical parameters, liver enzymes and damage of different tissues of male mice exposed to graphene oxide nanoparticles (GO NPs). In this research, 48 mice were divided into six groups. Hematology parameters including red blood cells (RBC), white blood cells (WBC), hematocrit (Ht), and hemoglobin (Hb), biochemical parameters such as cholesterol, triglyceride, glucose, protein, albumin, creatinine and bilirubin, liver enzymes including aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP) and lactate dehydrogenase (LDH) and histological damage of liver, kidney, lung and testis of mice were investigated. The results show that GO NPs in both concentrations (30 and 60 mg/kg/bw) decrease the number of RBC compared to the control group (p < 0.05). EAFE has no effect on RBC count while EAFE in combination with both concentrations of 30 and 60 mg/kg/bw GO NPs significantly (p < 0.05) improves the RBC count of mice. Also, the results show that EAFE improves the number of WBC, Ht, Hb, and biochemical parameters of mice exposed to different concentrations of GO NPs. A series of morphological changes were observed in the RBC of rats exposed to 60 mg/kg/bw GO NPs, including Dacrocyte, Poikilocyte and Schistocyte, which EAFE can improve the morphological changes caused in RBC. A series of histological damages such as degeneration and dilatation in the central vein in liver tissue, thickening and inflammatory cell infiltration in the renal capsule, peribronchiolar inflammatory cell infiltration and congestion in peribronchiolar blood vessels in lung tissue and decrease in the height of the germinal epithelium and a decrease in the population of spermatozoid cells in spermatogenic cells were observed in the testis of mice exposed to 60 mg/kg/bw GO NPs. The results show that EAFE can improve the damage caused by GO NPs in the liver, kidney and lung tissues, while it has no effect on the damage in the mice testis tissue. Therefore, GO NPs cause damage in the blood and different tissues of mice, and the use of EAFE can be effective in improving these damages.

High-Intensity Interval Training Induces Protein Lactylation in Different Tissues of Mice with Specificity and Time Dependence

Protein lysine lactylation (Kla) is a novel protein acylation reported in recent years, which plays an important role in the development of several diseases with pathologically elevated lactate levels, such as tumors. The concentration of lactate as a donor is directly related to the Kla level. High-intensity interval training (HIIT) is a workout pattern that has positive effects in many metabolic diseases, but the mechanisms by which HIIT promotes health are not yet clear. Lactate is the main metabolite of HIIT, and it is unknown as to whether high lactate during HIIT can induce changes in Kla levels, as well as whether Kla levels differ in different tissues and how time-dependent Kla levels are. In this study, we observed the specificity and time-dependent effects of a single HIIT on the regulation of Kla in mouse tissues. In addition, we aimed to select tissues with high Kla specificity and obvious time dependence for lactylation quantitative omics and analyze the possible biological targets of HIIT-induced Kla regulation. A single HIIT induces Kla in tissues with high lactate uptake and metabolism, such as iWAT, BAT, soleus muscle and liver proteins, and Kla levels peak at 24 h after HIIT and return to steady state at 72 h. Kla proteins in iWAT may affect pathways related to glycolipid metabolism and are highly associated with de novo synthesis. It is speculated that the changes in energy expenditure, lipolytic effects and metabolic characteristics during the recovery period after HIIT may be related to the regulation of Kla in iWAT.

Synergistic Effect Evaluation of Magnetotherapy and a Cationic-Magnetic Nanocomposite Loaded with Doxorubicin for Targeted Drug Delivery to Breast Adenocarcinoma.

This work investigates the synergistic effect of magnetotherapy and a novel cationic-magnetic drug delivery system on inhibiting breast cancer cell growth and other tissues. First, super-paramagnetic magnetite (Fe3O4) nanoparticles were coated with doxorubicin-imprinted poly(methacrylic acid-co-diallyl dimethylammonium chloride) [Fe3O4/poly(MAA-DDA)]. The cationic-magnetic nanocomposite (CMC) was characterized using XRD, FT-IR, VSM, TGA, TEM, FESEM, EDS, DLS, and BET. In vitro analyses, including drug release kinetics, cytotoxicity, and hemolytic assays, confirmed this novel CMC's good drug release profile and biocompatibility. Finally, in vivo experiments on BALB/c mice were designed to evaluate the synergistic effect of magnetotherapy on targeted drug delivery using the CMC. In vivo fluorescence imaging evaluated the drug distribution in different tissues of mice. Tumor volume evaluation demonstrated the efficiency of the CMC and magnetotherapy in preventing tumor growth; the two techniques significantly reduced tumor volume. Histopathological analysis proved that applying magnetotherapy in conjunction with the cationic-magnetic drug delivery system significantly prevented tumor cell proliferation and increased apoptosis with limited impact on other tissues. Also, Dox and Fe concentrations in different tissues confirmed the efficient drug delivery to tumor cells.

Complex genomic patterns of abasic sites in mammalian DNA revealed by a high-resolution SSiNGLe-AP method

DNA damage plays a critical role in biology and diseases; however, how different types of DNA lesions affect cellular functions is far from clear mostly due to the paucity of high-resolution methods that can map their locations in complex genomes, such as those of mammals. Here, we present the development and validation of SSiNGLe-AP method, which can map a common type of DNA damage, abasic (AP) sites, in a genome-wide and high-resolution manner. We apply this method to six different tissues of mice with different ages and human cancer cell lines. We find a nonrandom distribution of AP sites in the mammalian genome that exhibits dynamic enrichment at specific genomic locations, including single-nucleotide hotspots, and is significantly influenced by gene expression, age and tissue type in particular. Overall, these results suggest that we are only starting to understand the true complexities in the genomic patterns of DNA damage.

Accurate in vivo real-time determination of the hydrogen concentration in different tissues of mice after hydrogen inhalation

As an antioxidant, anti-inflammatory and anti-apoptotic agent, hydrogen (H2) shows a promising potential in basic and clinical research against various diseases owing to its safety and efficacy. However, knowledge involving its underlying mechanisms of action, dosage effects, and dose duration remains limited. Previously, the dynamics of H2 concentrations in different tissues of rats after exogenous H2 inhalation had been detected by our team. Here, sequential changes of H2 concentrations in different tissues of another most commonly used experimental rodent mice were monitored in real time with an electrochemical H2 gas sensor during continuous different concentrations of H2 inhalation targeting on five tissues including brain, liver, spleen, kidney, and gastrocnemius. The results showed that the H2 saturation concentrations varied among tissues significantly regardless of the concentration of H2 inhaled, and they were detected the highest in the kidney but the lowest in the gastrocnemius. Meantime, it required a significant longer time to saturate in the thigh muscle. By comparing the H2 saturation concentrations of mice and rats, we found that there were no differences detected in most tissues except the kidney and spleen. Both gas diffusion and bloodstream transport could help the H2 reach to most organs. The results provide data reference for dosage selection, dose duration determination to ensure optimal therapeutic effects of H2 for mice experiments.

Vertebrate extracellular matrix protein hemicentin-1 interacts physically and genetically with basement membrane protein nidogen-2

Hemicentins are large proteins of the extracellular matrix that belong to the fibulin family and play pivotal roles during development and homeostasis of a variety of invertebrate and vertebrate tissues. However, bona fide interaction partners of hemicentins have not been described as yet. Here, applying surface plasmon resonance spectroscopy and co-immunoprecipitation, we identify the basement membrane protein nidogen-2 (NID2) as a binding partner of mouse and zebrafish hemicentin-1 (HMCN1), in line with the formerly described essential role of mouse HMCN1 in basement membrane integrity. We show that HMCN1 binds to the same protein domain of NID2 (G3) as formerly shown for laminins, but with an approximately 3.5-fold lower affinity and in a competitive manner. Furthermore, immunofluorescence and immunogold labeling revealed that HMCN1/Hmcn1 is localized close to basement membranes and in partial overlap with NID2/Nid2a in different tissues of mouse and zebrafish. Genetic knockout and antisense-mediated knockdown studies in zebrafish further show that loss of Nid2a leads to similar defects in fin fold morphogenesis as the loss of Laminin-α5 (Lama5) or Hmcn1. Finally, combined partial loss-of-function studies indicated that nid2a genetically interacts with both hmcn1 and lama5. Together, these findings suggest that despite their mutually exclusive physical binding, hemicentins, nidogens, and laminins tightly cooperate and support each other during formation, maintenance, and function of basement membranes to confer tissue linkage.

VDR mediated HSD3B1 to regulate lipid metabolism and promoted testosterone synthesis in mouse Leydig cells.

The vitamin D receptor (VDR) mediates the pleiotropic biological actions that include osteoporosis, immune responses and androgen synthesis wherein the VDR transcriptionally regulates expression of the genes involved in this complex process. 3β-Hydroxysteroid dehydrogenase-1 (HSD3B1) is an absolutely necessary enzyme for androgen synthesis. The purpose of the present study was to explore the molecular mechanism of VDR mediated HSD3B1 regulation of lipid metabolism and testosterone synthesis. The levels of VDR, HSD3B1 and lipid metabolism associated protein were determined by quantitative real-time polymerase chain reaction (RT-qPCR) or western blot. The levels of testosterone concentrations in cell culture media serum by enzyme-linked immunosorbent assay (ELISA). Targeted relationship between VDR and Hsd3b1 was evaluated by dual-luciferase reporter assay. Based on the data analysis of mouse testicular proteome, we found that the expression of HSD3B1 was significantly reduced after VDR deletion. Here, we identified that Hsd3b1 was widely expressed in different tissues of mice by RT-qPCR, and was highly expressed in testis, and mainly located in testicular Leydig cells. Dual-luciferase assay confirmed that VDR could bind candidate vitamin D responsive elements (VDREs) in upstream region of Hsd3b1, and enhance gene expression. Furthermore, over-expression VDR and HSD3B1 significantly increased testosterone synthesis in mice Leydig cells. Meanwhile, Lpl expression was significantly down-regulated and Angptl4 expression was significantly up-regulated in the present of HSD3B1 overexpression. Both LPL and ANGPTL4 play important roles in regulating lipid metabolism. The present study unveiled VDR mediated HSD3B1 to regulate lipid metabolism and promoted testosterone synthesis in mouse Leydig cells. These findings will greatly help us to understand the roles of VDR and HSD3B1 in testosterone synthesis and lipid metabolism.

Characterization of Transverse Aortic Constriction in Mice Based on the Specific Recruitment of Leukocytes to the Hypertrophic Myocardium and the Aorta Ascendens.

Transverse aortic constriction (TAC) is a model that mimics pressure overload-induced left ventricular (LV) hypertrophy in mice. Alterations in immune cell functionality can promote cardiac and vascular remodeling. In the present study, we characterized the time course in innate immune cell dynamics in response to TAC in the different tissues of mice. It was determined whether TAC induces a characteristic leukocyte-driven immune response in the myocardium, aorta ascendens and descendens, spleen, blood, and draining lymph nodes supported by cytokine-driven chemotaxis in mice at 3, 6, and 21 days following surgery. We used complex flow cytometry staining combinations to characterize the various innate immune cell subsets and a multiplex array to determine cytokine concentrations in the serum. The results of the current study indicated that leukocytes accumulate in the myocardium and aorta ascendens in response to TAC. The leukocyte dynamics in the myocardium were dominated by the Ly6Clow macrophages with an early accumulation, whereas the response in the aorta ascendens was characterized by a long-lasting proinflammatory phenotype driven by Ly6Chigh macrophages, neutrophils, and activated DCs. In contrast to the high-pressure environment of the aorta ascendens, the tissue of the aorta descendens did not react to TAC with any leukocyte increase. The levels of proinflammatory cytokines in the blood were elevated in response to TAC, indicating a systemic reaction. Moreover, our findings strongly suggest that cardiac macrophages could origin from splenic pools and reach the site of the inflammation via the blood. Based on the current findings, it can be concluded that the high-pressure conditions in the aorta ascendens cause a characteristic immune response, dominated by the accumulation of leukocytes and the activation of DCs that varies in comparison to the immune cell dynamics in the myocardium and the aorta descendens.

Therapeutic effects of benznidazole in Swiss mice that are orally inoculated with Trypanosoma cruzi IV strains from the Western Brazilian Amazon

Strains of Trypanosoma cruzi, etiological agent of Chagas disease, are classified into different discrete typing units that may present distinct dynamics of infection and susceptibility to benznidazole (BZ) treatment. Mice that were orally inoculated with T. cruzi IV strains exhibited a more intense course of infection compared with intraperitoneally inoculated mice, reflected by higher parasite loads. We evaluated the efficacy of BZ treatment in Swiss mice that were inoculated with T. cruzi IV strains from the Western Brazilian Amazon. The mice were orally (OR) or intraperitoneally (IP) inoculated with 2 × 106 culture-derived metacyclic trypomastigotes of the AM14, AM16, AM64, and AM69 strains of T. cruzi that were obtained from two outbreaks of orally acquired acute Chagas disease in the state of Amazonas, Brazil. The animals were treated with BZ (100 mg/kg/day for 20 days). Fresh blood examination, hemoculture, conventional and quantitative real-time polymerase chain reaction were performed to monitor the therapeutic effects of BZ. Significant reductions in five of 24 parameters of parasitemia and parasite load were found in different tissues in the OR group, indicating worse response to BZ treatment compared with the IP group, in which significant reductions in nine of those 24 parameters were observed. The cure rates in the OR groups ranged from 18.2% (1/11) to 75.0% (9/12) and in the IP groups from 58.3% (7/12) to 91.7% (11/12), for the AM14 and AM69 strains, respectively. These findings indicate that treatment with BZ had fewer beneficial effects with regard to reducing parasitemia and parasite load in different tissues of mice that were OR inoculated with four TcIV strains compared with IP inoculation. Therefore, the route of infection with T. cruzi should be considered when evaluating the therapeutic efficacy of BZ in patients with Chagas disease.

In vivo toxicities of the hospital effluent in Mahdia Tunisia.

Hospital effluent (HE) is one of the most important sources of pharmaceuticals released into the environment. This kind of pollution is a recognized problem for both human health and aquatic life. Consequently, in the present study, we assessed the effects of untreated hospital effluent on mice via biochemical and histopathological determinations. Female mice were given free access to water bottles containing untreated HE at different dilutions for 21 days. Then clinical biochemistry and histopathology evaluation were conducted. Serum biochemistry analysis showed the presence of significant increase in cholesterol, triglycerides, glycaemia and total bilirubin. However, phosphatase alkaline and urea activities have been significantly decreased compared to the control group. No significant variation was observed for the rest of the studied parameters (high-density lipoproteins; low-density lipoproteins and uric acid). Additionally, multiple alterations, including cellular necrosis, leucocyte infiltration and congestion, were observed in different tissues of mice exposed to the tested HE.

Expression profiling of WD40 family genes including DDB1- and CUL4- associated factor (DCAF) genes in mice and human suggests important regulatory roles in testicular development and spermatogenesis

BackgroundThe WD40-repeat containing proteins, including DDB1–CUL4-associated factors (DCAFs), are abundant and conserved proteins that play important roles in different cellular processes including spermatogenesis. DCAFs are subset of WD40 family proteins that contain WDxR motif and have been proposed to function as substrate receptor for Cullin4-RING-based E3 ubiquitin ligase complexes to recruit diverse proteins for ubiquitination, a vital process in spermatogenesis. Large number of WD40 genes has been identified in different species including mouse and human. However, a systematic expression profiling of WD40 genes in different tissues of mouse and human has not been investigated. We hypothesize that large number of WD40 genes may express highly or specifically in the testis, where their expression is uniquely regulated during testis development and spermatogenesis. Therefore, the objective of this study is to mine and characterize expression patterns of WD40 genes in different tissues of mouse and human with particular emphasis on DCAF genes expressions during mouse testicular development.ResultsPublically available RNA sequencing (RNA seq) data mining identified 347 and 349 WD40 genes in mouse and human, respectively. Hierarchical clustering and heat map analyses of RNA seq datasets revealed differential expression patterns of WD40 genes with around 60–73% of the genes were highly or specifically expressed in testis. Similarly, around 74–83% of DCAF genes were predominantly or specifically expressed in testis. Moreover, WD40 genes showed distinct expression patterns during embryonic and postnatal testis development in mice. Finally, different germ cell populations of testis showed specific patterns of WD40 genes expression. Predicted gene ontology analyses revealed more than 80% of these proteins are implicated in cellular, metabolic, biological regulation and cell localization processes.ConclusionsWe have identified large number of WD40 family genes that are highly or specifically expressed in the testes of mouse and human. Moreover, WD40 genes have distinct expression patterns during embryonic and postnatal development of the testis in mice. Further, different germ cell populations within the testis showed specific patterns of WD40 genes expression. These results provide foundation for further research towards understanding the functional genomics and molecular mechanisms of mammalian testis development and spermatogenesis.

Salvia miltiorrhiza-Derived Sal-miR-58 Induces Autophagy and Attenuates Inflammation in Vascular Smooth Muscle Cells

Autophagy is associated with the cytoprotection of physiological processes against inflammation and oxidative stress. Salvia miltiorrhiza possesses cardiovascular protective actions and has powerful anti-oxidative and anti-inflammatory effects; however, whether and how Salvia miltiorrhiza-derived microRNAs (miRNAs) protect vascular smooth muscle cells (VSMCs) by inducing autophagy across species are unknown. We first screened and identified Sal-miR-58 from Salvia miltiorrhiza as a natural autophagy inducer. Synthetic Sal-miR-58 suppresses chronic angiotensin II (Ang II) infusion-induced abdominal aortic aneurysm (AAA) formation in mice, as well as induces autophagy in VSMCs and attenuates the inflammatory response elicited by Ang II in vivo and in vitro. Mechanistically, Sal-miR-58 downregulates Krüppel-like factor 3 (KLF3) expression through direct binding to the 3′ UTR of KLF3, which in turn relieves KLF3 repression of E3 ubiquitin ligase neural precursor cell-expressed developmentally downregulated 4-like (NEDD4L) expression, whereas NEDD4L upregulation increases the ubiquitination and degradation of the platelet isoform of phosphofructokinase (PFKP), subsequently leading to a decrease in the activation of Akt/mammalian target of rapamycin (mTOR) signaling and facilitating VSMC autophagy induced by Sal-miR-58 in the context of chronic Ang II stimulation and aneurysm formation. Our results provide the first evidence that plant-derived Sal-miR-58 induces autophagy and attenuates inflammation in VSMCs through cross-species modulation of the KLF3/NEDD4L/PFKP regulatory pathway.

Enterotoxic, neurotoxic and cytotoxice demonstrated by shiga toxin (2d) producing escherichia coli in experimental models

Background: Shiga toxin (Stx) producing Escherichia coli (STEC) colonize human intestinal tract and their infections have asymptomatic clinical manifestations which cause local and systemic pathological changes.
 Objectives: This study intended to establish the role of Shiga toxin (Stx2d) in developing clinical manifestations in STEC infections using experimental models.
 Methods: A total 300 stool samples were screened from hospitalised diarrhoeal patients enrolled in 2% surveillance system at International Centre for Diarrhoeal Disease Research, Bangladesh (icddr,b). The stx gene profile including their variants was identified byPCR.stx2d gene positive STEC PT187 was selected for toxin (s) preparation.Toxin was prepared by centrifugation of culture supernatant. Enterotoxicand paralytic-lethal activities were tested in rabbit ileal loops and mice, respectively. Histopathological study of the rabbit ileal loop segments and different tissues of mice by paraffin embedded method and stained by H & E staining. Cytotoxic effect was performed on HeLa cells.
 Results: Nine STEC strains were identified for stx2 gene positive. Among them STEC PT187 was found stx2d gene positive strain and selected for toxic activities. Toxin (s) responsible for causing accumulation of fluid in rabbit ileal loops and its segments showed inflammation and enterocytenecrosis. In mouse model, toxin(s) was found to cause hind limbs paralysis and death. Brain, spinal cord and kidney tissue of mice showed histopathological changes. Toxin (s) also showed positive cytotoxic activity in HeLa cell.
 Conclusion: In this study, results indicated that Stx2d producing E. coli exhibit not only enterotoxic activity, but also cause impaired neurological functions and cytotoxic effect.
 Bangladesh Med Res Counc Bull 2020; 46(1): 41-47

Expressions and significances of the angiotensin-converting enzyme 2 gene, the receptor of SARS-CoV-2 for COVID-19.

The ACE2 gene is a receptor of SARS-CoV-2 (severe acute respiratory syndrome coronavirus 2) for COVID-19 (coronavirus disease 2019). To analyze the expression profiles and clinical significances for this gene in humans, RNA-seq data representing 27 different tissues were analyzed using NCBI; total RNA was extracted from different tissues of mouse and semi-quantitative reverse transcriptional-polymerase chain reaction (Q-RT-PCR) was carried out. Immunohistochemistry expression profiles in normal tissues and cancer tissues and TCGA survival analysis in renal and liver cancer were conducted. ACE2 was highly conserved in different species. In normal tissues, ACE2 expression distributions were organ-specific, mainly in the kidney, male testis and female breast, and cardiovascular and gastrointestinal systems. High level of expression in testis, cardiovascular and gastrointestinal system indicated that SARS-CoV-2 might not only attack the lungs, but also affect other organs, particularly the testes, thus it may severely damage male sexual development for younger male and lead to infertility in an adult male, if he contracted COVID-19. On the other side, high expression of ACE2 was correlated with increased survival rate in renal and liver cancer, indicating that ACE2 is a prognostic marker in both renal cancer and liver cancers. Thus, the ACE2 is a functional receptor for SARS-CoV-2 and has a potential anti-tumor role in cancer. Taken together, this study may not only provide potential clues for further medical pathogenesis of COVID-19 and male fertility, but also indicate the clinical significance of the role of the ACE2 gene in cancer.

Dyno to use machine learning for AAV design

Adeno-associated viruses (AAVs) are the DNA-delivery vehicle of choice for many gene-therapy companies, but they’re not perfect. Many past attempts to engineer improved AAVs have failed, because the protein shell of the virus—the capsid—is like a Rubik’s Cube. Improving one feature often throws others out of line. A new study suggests that machine learning could help solve that molecular puzzle. And a new start-up, Dyno Therapeutics, has been quietly raising money to test the concept on an industrial scale. A team led by George Church at Harvard Medical School created all possible single codon mutations—substitutions, deletions, and insertions—in the capsid of an AAV variant called AAV2. The team then tested how the mutations altered the AAVs’ immunogenicity, thermostability, ability to multiply in cells, and distribution to different tissues in mice. With all this data in hand, the team set out to introduce multiple mutations in AAVs to improve their delivery

Characterization, evaluation of nutritional parameters of Radix isatidis protein and its antioxidant activity in D-galactose induced ageing mice

BackgroundRadix isatidis (Isatis indigotica Fort.) is an ancient medicinal herb, which has been applied to the prevention and treatment of influenza virus since ancient times. In recent years, the antioxidant activity of Radix isatidis has been widely concerned by researchers. Our previous studies have shown that Radix isatidis protein (RIP) has good antioxidant activity in vitro. In this study, the composition of the protein was characterized and its antioxidant activity in vivo was evaluated.MethodsThe model of oxidative damage in mice was established by subcutaneous injection of D-galactose for 7 weeks. Commercially available kits were used to determine the content of protein and several oxidation indexes in different tissues of mice. The tissue samples were stained with hematoxylin and eosin (H&E) and the pathological changes were observed by optical microscope. The molecular weight of RIP was analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The amino acid composition of RIP was determined by a non-derivative method developed by our research group.ResultsRIP significantly increased the activities of antioxidant enzymes such as SOD, CAT, GSH-Px and total antioxidant capability (TAOC) but decreased the MDA level in the serum, kidney and liver. H&E stained sections of liver and kidney revealed D-galactose could cause serious injury and RIP could substantially attenuate the injury. The analysis of SDS-PAGE showed that four bands with molecular weights of 19.2 kDa, 21.5 kDa, 24.8 kDa and 40.0 kDa were the main protein components of RIP.ConclusionsThe results suggested that RIP had excellent antioxidant activity, which could be explored as a health-care product to retard aging and a good source of protein nutrition for human consumption.

A novel assay for detection and quantification of C-mannosyl tryptophan in normal or diabetic mice

C-Mannosyl tryptophan (C-Man-Trp) is a unique molecule in that an α-mannose is connected to the indole C2 carbon atom of a Trp residue via C-glycosidic linkage. Although serum C-Man-Trp may be a novel biomarker of renal function in humans, the biological significance of C-Man-Trp has yet to be fully investigated. In this study, a novel assay system for C-Man-Trp was established using hydrophilic-interaction liquid chromatography, followed by detecting the fluorescence intensity or mass abundance of C-Man-Trp. Using this system, we systematically assessed the amount of free monomeric C-Man-Trp in different tissues of mice. The tissue level of C-Man-Trp was high, especially in the ovaries and uterus. Other organs with high levels of C-Man-Trp included the brain, spleen, lungs, bladder, and testes. The level was low in skeletal muscle. We also investigated whether the tissue level of C-Man-Trp is affected in diabetes. In KK-Ay diabetic mice, the level of urinary C-Man-Trp excretion was increased, and the tissue levels of C-Man-Trp were decreased in the liver but increased in the kidney. These results demonstrate that C-Man-Trp is differentially distributed in numerous tissues and organs in mice, and the levels are altered by disordered carbohydrate metabolism such as diabetes.