Prognostic value of plasma big endothelin-1 in patients with light chain cardiac amyloidosis.

BACKGROUND: Light chain cardiac amyloidosis (AL-CA) is associated with a high incidence of mortality. Big endothelin-1 (ET-1), the precursor of endothelial-vasoconstrictive ET-1, is closely related to the concentration of bioactive ET-1. Association between big ET-1 and prognosis of AL-CA has not yet been documented. The purpose of this study was to evaluate the prognostic value of big ET-1 for poor outcomes in moderate to severe AL-CA. METHODS: Big ET-1 levels were determined on admission in patients with newly diagnosed AL-CA with modified Mayo 2004 stage II or III. Primary outcome was all-cause mortality. The secondary outcomes included death from cardiac cause and the composite of the primary outcome or hospitalisations due to worsening heart failure. RESULTS: Overall, 141 patients were retrospectively included (57 stage II, 34 stage IIIa, 50 stage IIIb). During a median follow-up time of 25.7 months, 84 (59.6%) patients died. Patients with big ET-1 levels of ≤0.88 pmol/L had longer survival than those with >0.88 pmol/L (median survival time: 34.1 months vs 15.3 months, log-rank p<0.001), which was also observed in the validation cohort (log-rank p=0.026). Higher big ET-1 levels were predictive for all-cause mortality after multivariable adjustment (HR 1.91, 95% CI 1.05 to 3.49, p=0.035). Big ET-1 levels added an incremental prognostic value over modified Mayo 2004 stage (C-index: from 0.671 to 0.696, p=0.025; integrated discrimination improvement 0.168, p=0.047). CONCLUSIONS: Big ET-1 is a strong and independent predictor of mortality in patients with moderate to severe AL-CA, which may indicate a possible role for risk stratification in patients with this disease.

Prognostic implications of premature ventricular contractions and non-sustained ventricular tachycardia in light-chain cardiac amyloidosis.

AIMS: Premature ventricular contractions (PVC) and non-sustained ventricular tachycardia (NSVT) are commonly observed in light chain cardiac amyloidosis (AL-CA), but their association with prognosis is still unclear. We aimed to evaluate the prognostic value of PVCs and NSVT in patients with moderate-to-advanced AL-CA. METHODS AND RESULTS: We retrospectively included patients with AL-CA at modified 2004 Mayo stages II-IIIb between February 2014 and December 2020. Twenty-four-hour Holter recordings were assessed on admission. The outcomes included (i) new onset of adverse ventricular arrhythmia (VA) or sudden cardiac death (SCD) and (ii) cardiac death during follow-up. Of the 143 patients studied (60.41 ± 11.06 years, male 64.34%), 132 (92.31%) had presence of PVC, and 50 (34.97%) had NSVT on Holter. Twelve (8.4%) patients died in hospital and 131 patients were followed up (median 24.4 months), among whom 71 patients had cardiac death, and 15 underwent adverse VA/SCD. NSVT [hazard ratio (HR): 13.57, 95% confidence interval (CI): 3.06-60.18, P < 0.001], log-transformed PVC counts (HR: 1.46, 95%CI: 1.15-1.86, P = 0.002) and PVC burden (HR: 1.43 95%CI:1.14-1.80, P = 0.002) were predictive of new onset of adverse VA/SCD. The highest tertile of PVC counts (HR: 2.33, 95%CI: 1.27-4.28, P = 0.006) and PVC burden (HR: 2.58, 95%CI: 1.42-4.69, P = 0.002), rather than NSVT (HR: 1.16, 95%CI: 0.67-1.98, P = 0.603), was associated with cardiac death. Higher PVC counts/burden provided incremental value on modified 2004 Mayo stage in predicting cardiac death, with C index increasing from 0.681 to 0.712 and 0.717, respectively (P values <0.05). CONCLUSION: PVC count, burden, and NSVT significantly correlated with adverse VA/SCD during follow-up in patients with AL-CA. Higher PVC counts/burdens added incremental value for predicting cardiac death.

The chromosome-level assembly of the wild diploid alfalfa genome provides insights into the full landscape of genomic variations between cultivated and wild alfalfa.

Alfalfa (Medicago sativa L.) is one of the most important forage legumes in the world, including autotetraploid (M. sativa ssp. sativa) and diploid alfalfa (M. sativa ssp. caerulea, progenitor of autotetraploid alfalfa). Here, we reported a high-quality genome of ZW0012 (diploid alfalfa, 769 Mb, contig N50 = 5.5 Mb), which was grouped into the Northern group in population structure analysis, suggesting that our genome assembly filled a major gap among the members of M. sativa complex. During polyploidization, large phenotypic differences occurred between diploids and tetraploids, and the genetic information underlying its massive phenotypic variations remains largely unexplored. Extensive structural variations (SVs) were identified between ZW0012 and XinJiangDaYe (an autotetraploid alfalfa with released genome). We identified 71 ZW0012-specific PAV genes and 1296 XinJiangDaYe-specific PAV genes, mainly involved in defence response, cell growth, and photosynthesis. We have verified the positive roles of MsNCR1 (a XinJiangDaYe-specific PAV gene) in nodulation using an Agrobacterium rhizobia-mediated transgenic method. We also demonstrated that MsSKIP23_1 and MsFBL23_1 (two XinJiangDaYe-specific PAV genes) regulated leaf size by transient overexpression and virus-induced gene silencing analysis. Our study provides a high-quality reference genome of an important diploid alfalfa germplasm and a valuable resource of variation landscape between diploid and autotetraploid, which will facilitate the functional gene discovery and molecular-based breeding for the cultivars in the future.

Spiking Neural Network Based on Multi-Scale Saliency Fusion for Breast Cancer Detection.

Deep neural networks have been successfully applied in the field of image recognition and object detection, and the recognition results are close to or even superior to those from human beings. A deep neural network takes the activation function as the basic unit. It is inferior to the spiking neural network, which takes the spiking neuron model as the basic unit in the aspect of biological interpretability. The spiking neural network is considered as the third-generation artificial neural network, which is event-driven and has low power consumption. It modulates the process of nerve cells from receiving a stimulus to firing spikes. However, it is difficult to train spiking neural network directly due to the non-differentiable spiking neurons. In particular, it is impossible to train a spiking neural network using the back-propagation algorithm directly. Therefore, the application scenarios of spiking neural network are not as extensive as deep neural network, and a spiking neural network is mostly used in simple image classification tasks. This paper proposed a spiking neural network method for the field of object detection based on medical images using the method of converting a deep neural network to spiking neural network. The detection framework relies on the YOLO structure and uses the feature pyramid structure to obtain the multi-scale features of the image. By fusing the high resolution of low-level features and the strong semantic information of high-level features, the detection precision of the network is improved. The proposed method is applied to detect the location and classification of breast lesions with ultrasound and X-ray datasets, and the results are 90.67% and 92.81%, respectively.

A Parallel Spiking Neural Network Based on Adaptive Lateral Inhibition Mechanism for Objective Recognition.

Spiking neural network (SNN) has attracted extensive attention in the field of machine learning because of its biological interpretability and low power consumption. However, the accuracy of pattern recognition cannot completely surpass deep neural networks (DNNs). The main reason is that the inherent nondifferentiability of spiking neurons makes SNN unable to be trained directly by the gradient descent algorithm, and there is also no unified training algorithm for SNN. Inspired by the biological vision system, this paper proposes a parallel convolution SNN structure combined with an adaptive lateral inhibition mechanism. And, a way of dynamically evolving the time constant with the training of SNN is proposed to ensure the diversity of neurons. This paper verifies the effectiveness of the proposed methods on static datasets and neuromorphic datasets and extends it to the recognition of breast tumors. Experimental results show that the SNN has obvious advantages in dynamical datasets. For breast tumors, it is also an edge-based task, because the edge of a medical image contains the most important information in the image. This kind of information can provide great help for the noninvasive and accurate diagnosis of diseases. The Experimental results show that the proposed method is very close to the recognition results of DNNs on static datasets, and its performance on neuromorphic datasets exceeds that of DNNs.

Chronic Glutathione Depletion Confers Protection against Alcohol-induced Steatosis: Implication for Redox Activation of AMP-activated Protein Kinase Pathway.

The pathogenesis of alcoholic liver disease (ALD) is not well established. However, oxidative stress and associated decreases in levels of glutathione (GSH) are known to play a central role in ALD. The present study examines the effect of GSH deficiency on alcohol-induced liver steatosis in Gclm knockout (KO) mice that constitutively have ≈15% normal hepatic levels of GSH. Following chronic (6 week) feeding with an ethanol-containing liquid diet, the Gclm KO mice were unexpectedly found to be protected against steatosis despite showing increased oxidative stress (as reflected in elevated levels of CYP2E1 and protein carbonyls). Gclm KO mice also exhibit constitutive activation of liver AMP-activated protein kinase (AMPK) pathway and nuclear factor-erythroid 2-related factor 2 target genes, and show enhanced ethanol clearance, altered hepatic lipid profiles in favor of increased levels of polyunsaturated fatty acids and concordant changes in expression of genes associated with lipogenesis and fatty acid oxidation. In summary, our data implicate a novel mechanism protecting against liver steatosis via an oxidative stress adaptive response that activates the AMPK pathway. We propose redox activation of the AMPK may represent a new therapeutic strategy for preventing ALD.

Update of the human and mouse Fanconi anemia genes.

Fanconi anemia (FA) is a recessively inherited disease manifesting developmental abnormalities, bone marrow failure, and increased risk of malignancies. Whereas FA has been studied for nearly 90 years, only in the last 20 years have increasing numbers of genes been implicated in the pathogenesis associated with this genetic disease. To date, 19 genes have been identified that encode Fanconi anemia complementation group proteins, all of which are named or aliased, using the root symbol "FANC." Fanconi anemia subtype (FANC) proteins function in a common DNA repair pathway called "the FA pathway," which is essential for maintaining genomic integrity. The various FANC mutant proteins contribute to distinct steps associated with FA pathogenesis. Herein, we provide a review update of the 19 human FANC and their mouse orthologs, an evolutionary perspective on the FANC genes, and the functional significance of the FA DNA repair pathway in association with clinical disorders. This is an example of a set of genes--known to exist in vertebrates, invertebrates, plants, and yeast--that are grouped together on the basis of shared biochemical and physiological functions, rather than evolutionary phylogeny, and have been named on this basis by the HUGO Gene Nomenclature Committee (HGNC).

Transgenic mouse models for alcohol metabolism, toxicity, and cancer.

Alcohol abuse leads to tissue damage including a variety of cancers; however, the molecular mechanisms by which this damage occurs remain to be fully understood. The primary enzymes involved in ethanol metabolism include alcohol dehydrogenase (ADH), cytochrome P450 isoform 2E1, (CYP2E1), catalase (CAT), and aldehyde dehydrogenases (ALDH). Genetic polymorphisms in human genes encoding these enzymes are associated with increased risks of alcohol-related tissue damage, as well as differences in alcohol consumption and dependence. Oxidative stress resulting from ethanol oxidation is one established pathogenic event in alcohol-induced toxicity. Ethanol metabolism generates free radicals, such as reactive oxygen species (ROS) and reactive nitrogen species (RNS), and has been associated with diminished glutathione (GSH) levels as well as changes in other antioxidant mechanisms. In addition, the formation of protein and DNA adducts associated with the accumulation of ethanol-derived aldehydes can adversely affect critical biological functions and thereby promote cellular and tissue pathology. Animal models have proven to be valuable tools for investigating mechanisms underlying pathogenesis caused by alcohol. In this review, we provide a brief discussion on several animal models with genetic defects in alcohol-metabolizing enzymes and GSH-synthesizing enzymes and their relevance to alcohol research.

Mitochondrial targeting of mouse NQO1 and CYP1B1 proteins.

Four dioxin-inducible enzymes--NAD(P)H: quinone oxidoreductase-1 (NQO1) and three cytochromes P450 (CYP1A1, CYP1A2 & CYP1B1)--are implicated in both detoxication and metabolic activation of various endobiotics and xenobiotics. NQO1 is generally regarded as a cytosolic enzyme; whereas CYP1 proteins are located primarily in endoplasmic reticulum (ER), CYP1A1 and CYP1A2 proteins are also targeted to mitochondria. This lab has generated Cyp1a1(mc/mc) and Cyp1a1(mtt/mtt) knock-in mouse lines in which CYP1A1 protein is targeted exclusively to ER (microsomes) and mitochondria, respectively. Comparing dioxin-treated Cyp1(+/+) wild-type, Cyp1a1(mc/mc), Cyp1a1(mtt/mtt), and Cyp1a1(-/-), Cyp1b1(-/-) and Nqo1(-/-) knockout mice, in the present study we show that [a] NQO1 protein locates to cytosol, ER and mitochondria, [b] CYP1B1 protein (similar to CYP1A1 and CYP1A2 proteins) traffics to mitochondria as well as ER, and [c] NQO1 and CYP1B1 targeting to mitochondrial or ER membranes is independent of CYP1A1 presence in that membrane.

Generation of a Slc39a8 hypomorph mouse: markedly decreased ZIP8 Zn²âº/(HCO3⁻)2 transporter expression.

Previously this laboratory has identified the mouse Slc39a8 gene encoding the ZIP8 transporter, important in cadmium uptake. ZIP8 functions endogenously as a electroneutral Zn(2+)/(HCO(3)(-))(2) symporter, moving both ions into the cell. The overall physiological importance of ZIP8 remains unclear. Herein we describe generation of a mouse line carrying the Slc39a8(neo) allele, containing the Frt-flanked neomycin-resistance (neo) mini-cassette in intron 3 and loxP sites in introns 3 and 6. Cre recombinase functions correctly in Escherichia coli and in adeno-Cre-infected mouse fetal fibroblasts, but does not function in the intact mouse for reasons not clear. Slc39a8(neo) is a hypomorphic allele, because Slc39a8(neo/neo) homozygotes exhibit dramatically decreased ZIP8 expression in embryo, fetus, and visceral yolk sac - in comparison to their littermate wild-type controls. This ZIP8 hypomorph will be instrumental in studying developmental and in utero physiological functions of the ZIP8 transporter.

[Effects of hypoxia on the secretions of proinflammatory cytokines TNF-alpha and IL-6 in macrophages and its mechanism].

AIM: To investigate the effects of hypoxia on the secretions of proinflammatory cytokines TNF-alpha and IL-6 and to inquire into the mechanism. METHODS: Separated mice abdominal macrophages which were identified with non-specific esterase dye method, and created the hypoxic cultured model. The levels of TNF-alpha and IL-6 in the medium were determined by ELISA method. The mRNA expressions of TNF-alpha and IL-6 were measured by RT-PCR method. NF-kappaB activation was assayed by Western blot method. Finaly, we added cortone (5 microg/ml) to the medium, then observed the secretion levels of TNF-alpha and IL-6 during hypoxia. RESULTS: The secretions of TNF-alpha and IL-6 from Mphi exposed to hypoxia for 12 h were increased significantly compared with control (P < 0.01). The expressions of TNF-alpha mRNA and IL-6 mRNA were enhanced obviously contrasted with control (P < 0.01). NF-kappaB activation in Mphi nuclei was raised at 2 h during hypoxia and persisted to 5 h. We added cortone to the medium and found no significant change in secretion of TNF-alpha and IL-6 during hypoxia. CONCLUSION: Hypoxia could activate NF-kappaB and make it shift to nucleus which promoted the transcriptions and expressions of TNF-alpha and IL-6.

[Changes in expression of apoptosis correlative proteins and HIF-1 during acute hypoxia and hypoxic preconditioning in cultured cardiomyocyte of rat].

AIM: To investigate the relationship between the hypoxia-inducible factor 1(HIF-1) and apoptosis correlative proteins in the cardiomyocyte during hypoxia. METHODS: Rat cardiomyocytes cultured in vitro were divided into normoxia, hypoxia and hypoxic preconditioning groups. The cardiomyocytes in hypoxic preconditioning group were cultured in 1% O2, 5% CO2, 94% N2 for 5 days, 12 h daily before exposed to 0% O2 hypoxia with the hypoxic group. The protein expression of HIF-1alpha, Bcl-2, P53 and Bax in the cardiomyocytes were analysis with Western blot after 48 h 0% O2 hypoxia. RESULTS: With the increment of HIF-1 expression, the Bcl-2 expression was inhibited, the Bax and P53 expression were increased as well during hypoxia. Hypoxic preconditioning could suppress the HIF-1 expression, meanwhile the Bcl-2 expression was elevated, and the Bax expression was decreased. CONCLUSION: HIF-1 may contribute to the regulation of the cardiomyocyte apoptosis with the Bcl-2 family during hypoxia.