Mapping chromosomal instability induced by small-molecular therapeutics in a yeast model.

The yeast Saccharomyces cerevisiae has been widely used as a model system for studying the physiological and pharmacological action of small-molecular drugs. Here, a heterozygous diploid S. cerevisiae strain QSS4 was generated to determine whether drugs could induce chromosomal instability by determining the frequency of mitotic recombination. Using the combination of a custom SNP microarray and yeast screening system, the patterns of chromosomal instability induced by drugs were explored at the whole genome level in QSS4. We found that Zeocin (a member of the bleomycin family) treatment increased the rate of genomic alterations, including aneuploidy, loss of heterozygosity (LOH), and chromosomal rearrangement over a hundred-fold. Most recombination events are likely to be initiated by DNA double-stand breaks directly generated by Zeocin. Another remarkable finding is that G4-motifs and low GC regions were significantly underrepresented within the gene conversion tracts of Zeocin-induced LOH events, indicating that certain DNA regions are less preferred Zeocin-binding sites in vivo. This study provides a novel paradigm for evaluating genetic toxicity of small-molecular drugs using yeast models.

[Impacts of Climate Change and Human Activities on Vegetation Restoration in Typical Grasslands of China].

Grasslands, as one of the key ecosystems relevant to the terrestrial ecosystem carbon and water cycles as well as the ecological security in China, are very sensitive to climate change and human activities. However, the relative contributions of climate change and human activities on the vegetation restoration in those regions are still controversial. Using ecosystem net primary production (NPP) as an ecological indicator, this study quantified the relative roles of climate change and human activities on vegetation restoration in Chinese typical grasslands (northern temperate grasslands and Qinghai-Tibet Plateau alpine grasslands) by comparing the trends of actual NPP derived from MODIS and potential NPP estimated by the Thornthwaite Memorial model during 2000-2020. The results showed that approximately 93% of the grasslands in the study area experienced a recovering tendency, with an average increase of NPP (carbon) by 2.12 g·(m2·a)-1(P<0.01). Therein, nearly half of the vegetation-restored areas were jointly-dominated by climate change and human activities, whereas approximately 36% and 10% of the restored areas were controlled individually by climate change and human activities, respectively. In addition, the share of climate-change dominated areas differed greatly by grassland types, characterized by a much larger area percentage in the alpine grasslands than that in the temperate grasslands and an increasing area share with a drying background climate. This study suggested that human activities were not primarily responsible for the vegetation restoration in northern temperate grasslands and Qinghai-Tibet Plateau alpine grasslands, but they could decrease and even cancel the possible vegetation degeneration caused by worsening climate in a few regions. Long-term monitoring of vegetation dynamics and a multi-method comparison are needed in future studies.

Reduced expression of APLP2 in spinal GABAergic inhibitory neurons contributed to nerve injury-induced microglial activation and pain sensitization.

The amyloid precursor protein (APP) is critical for the pathogenesis of Alzheimer's disease (AD). The AD patients usually have lower pain sensitivity in addition to cognitive impairments. However, considerably less is known as yet about the role of APP and its two mammalian homologues, amyloid precursor-like protein 1 and 2 (APLP1, APLP2), in spinal processing of nociceptive information. Here we found that all APP family members were present in spinal cord dorsal horn of adult male C57BL/6J mice. Peripheral nerve injury specifically reduced the expression of spinal APLP2 that correlated with neuropathic mechanical allodynia. The loss of APLP2 was confined to inhibitory GABAergic interneurons. Targeted knockdown of APLP2 in GABAergic interneurons of GAD2-Cre mice evoked pain hypersensitivity by means of microglia activation. Our data showed that GABAergic terminals expressed APLP2, a putative cell adhesion protein that interacted with microglia-specific integrin molecule CD11b. Knocking down APLP2 in GAD2-positive neurons to disrupt the trans-cellular interaction led to microglia-dependent pain sensitization. Our data thus revealed an important role of APLP2 for GABAergic interneurons to control microglial activity and pain sensitivity.

Analysis of volatile organic compounds and potential odour compounds in food contact paperboard using headspace two-dimensional GC-QTOF-MS.

The objective was to establish a robust and reliable approach for the characterisation of volatile organic compounds (VOCs) present in food contact paperboard. This was achieved through the utilisation of headspace solid-phase microextraction in tandem with comprehensive two-dimensional (2D) gas chromatography (GC) and quadrupole time-of-flight mass spectrometry (HS-SPME-GC × GC-QTOF-MS). The experimental parameters were optimised, involving the use of a DVB/C-WR/PDMS fibre at a temperature of 80 °C for a duration of 30 min. A total of 344 VOCs comprising aldehydes, ketones, alcohols, ethers, esters, alkanes and aromatic compounds, were tentatively identified in the samples. Twelve compounds believed to be from biogenic sources had a high odour impact making them major contributors to potential taint from the paperboard samples. Significant attention should be devoted to five compounds namely, 2-methylnaphthalene, 2-pentyl-furan, furfural, 1-octen-3-one and 1-octen-3-ol due to their potential adverse impact on the organoleptic qualities of packaged food items and their potential toxicity.Abbreviations: C-WR: carbon wide range; DVB: divinylbenzene; GC-MS: gas chromatography - mass spectrometry; GCxGC-QTOF-MS: comprehensive two-dimensional gas chromatography coupled to quadrupole-time-of-flight - mass spectrometry; HS-SPME: headspace - solid phase microextraction; LOD: limit of detection; LOQ: limit of quantification; OAV: odor activity values; PDMS: polydimethylsiloxane; RI: retention index; TTC: threshold of toxicological concern; VOC: volatile organic compound.

Dual Role of Caspase 8 in Adipocyte Apoptosis and Metabolic Inflammation.

Caspases are cysteine-aspartic proteases that were initially discovered to play a role in apoptosis. However, caspase 8, in particular, also has additional nonapoptotic roles, such as in inflammation. Adipocyte cell death and inflammation are hypothesized to be initiating pathogenic factors in type 2 diabetes. Here, we examined the pleiotropic role of caspase 8 in adipocytes and obesity-associated insulin resistance. Caspase 8 expression was increased in adipocytes from mice and humans with obesity and insulin resistance. Treatment of 3T3-L1 adipocytes with caspase 8 inhibitor Z-IETD-FMK decreased both death receptor-mediated signaling and targets of nuclear factor κ-light-chain-enhancer of activated B (NF-κB) signaling. We generated novel adipose tissue and adipocyte-specific caspase 8 knockout mice (aP2Casp8-/- and adipoqCasp8-/-). Both males and females had improved glucose tolerance in the setting of high-fat diet (HFD) feeding. Knockout mice also gained less weight on HFD, with decreased adiposity, adipocyte size, and hepatic steatosis. These mice had decreased adipose tissue inflammation and decreased activation of canonical and noncanonical NF-κB signaling. Furthermore, they demonstrated increased energy expenditure, core body temperature, and UCP1 expression. Adipocyte-specific activation of Ikbkb or housing mice at thermoneutrality attenuated improvements in glucose tolerance. These data demonstrate an important role for caspase 8 in mediating adipocyte cell death and inflammation to regulate glucose and energy homeostasis. ARTICLE HIGHLIGHTS: Caspase 8 is increased in adipocytes from mice and humans with obesity and insulin resistance. Knockdown of caspase 8 in adipocytes protects mice from glucose intolerance and weight gain on a high-fat diet. Knockdown of caspase 8 decreases Fas signaling, as well as canonical and noncanonical nuclear factor κ-light-chain-enhancer of activated B (NF-κB) signaling in adipose tissue. Improved glucose tolerance occurs via reduced activation of NF-κB signaling and via induction of UCP1 in adipocytes.

Cbl-b modulated TrkA ubiquitination and function in the dorsal root ganglion of mice.

Casitas B-lineage lymphoma b (Cbl-b) is one of the E3 ubiquitin ligases that ubiquitinate Tropomyosin-related kinase A (TrkA), a key nerve growth factor receptor involved in the pathological pain. Here we found that Cbl-b was abundant in dorsal root ganglion (DRG) neurons of mice and co-localized with TrkA. Ubiquitination of TrkA by Cbl-b exerted a tonic negative control over the protein level of TrkA. Knockdown of Cbl-b caused TrkA accumulation in DRGs and evoked mechanical and heat hypersensitivity in intact mice. Our data showed that knee osteoarthritis induced by destabilization of the medial meniscus (DMM) led to the dissociation of Cbl-b with TrkA in DRG neurons, which impaired the ability of Cbl-b to ubiquitinate TrkA and served as an important mechanism to cause TrkA-dependent pain sensitization. Viral expression of constitutively active Cbl-b in DRGs of osteoarthritic mice effectively repressed TrkA protein level and more importantly, alleviated mechanical allodynia and heat hyperalgesia. Viral delivery of Cbl-b through intra-articular route generated a similar analgesic action. These data suggested that ubiquitination of TrkA by Cbl-b might represent an effective way to treat the osteoarthritic pain.

Macrophage Jak2 deficiency accelerates atherosclerosis through defects in cholesterol efflux.

Atherosclerosis is a chronic inflammatory condition in which macrophages play a major role. Janus kinase 2 (JAK2) is a pivotal molecule in inflammatory and metabolic signaling, and Jak2V617F activating mutation has recently been implicated with enhancing clonal hematopoiesis and atherosclerosis. To determine the essential in vivo role of macrophage (M)-Jak2 in atherosclerosis, we generate atherosclerosis-prone ApoE-null mice deficient in M-Jak2. Contrary to our expectation, these mice exhibit increased plaque burden with no differences in macrophage proliferation, recruitment or bone marrow clonal expansion. Notably, M-Jak2-deficient bone marrow derived macrophages show a significant defect in cholesterol efflux. Pharmacologic JAK2 inhibition with ruxolitinib also leads to defects in cholesterol efflux and accelerates atherosclerosis. Liver X receptor agonist abolishes the efflux defect and attenuates the accelerated atherosclerosis that occurs with M-Jak2 deficiency. Macrophages of individuals with the Jak2V617F mutation show increased efflux which is normalized when treated with a JAK2 inhibitor. Together, M-Jak2-deficiency leads to accelerated atherosclerosis primarily through defects in cholesterol efflux from macrophages.

Hepatic Igf1-Deficiency Protects Against Atherosclerosis in Female Mice.

Atherosclerosis is the leading cause of cardiovascular disease (CVD), with distinct sex-specific pathogenic mechanisms that are poorly understood. Aging, a major independent risk factor for atherosclerosis, correlates with a decline in circulating insulin-like growth factor-1 (IGF-1). However, the precise effects of Igf1 on atherosclerosis remain unclear. In the present study, we assessed the essential role of hepatic Igf1, the major source of circulating IGF-1, in atherogenesis. We generated hepatic Igf1-deficient atherosclerosis-prone apolipoprotein E (ApoE)-null mice (L-Igf1-/-ApoE-/-) using the Cre-loxP system driven by the Albumin promoter. Starting at 6 weeks of age, these mice and their littermate controls, separated into male and female groups, were placed on an atherogenic diet for 18 to 19 weeks. We show that hepatic Igf1-deficiency led to atheroprotection with reduced plaque macrophages in females, without significant effects in males. This protection from atherosclerosis in females was associated with increased subcutaneous adiposity and with impaired lipolysis. Moreover, this impaired lipid homeostasis was associated with disrupted adipokine secretion with reduced circulating interleukin-6 (IL-6) levels. Together, our data show that endogenous hepatic Igf1 plays a sex-specific regulatory role in atherogenesis, potentially through athero-promoting effects of adipose tissue-derived IL-6 secretion. These data provide potential novel sex-specific mechanisms in the pathogenesis of atherosclerosis.

Metabolic Role of PTEN in Insulin Signaling and Resistance.

Phosphatase and tensin homolog (PTEN) is most prominently known for its function in tumorigenesis. However, a metabolic role of PTEN is emerging as a result of its altered expression in type 2 diabetes (T2D), which results in impaired insulin signaling and promotion of insulin resistance during the pathogenesis of T2D. PTEN functions in regulating insulin signaling across different organs have been identified. Through the use of a variety of models, such as tissue-specific knockout (KO) mice and in vitro cell cultures, PTEN's role in regulating insulin action has been elucidated across many cell types. Herein, we will review the recent advancements in the understanding of PTEN's metabolic functions in each of the tissues and cell types that contribute to regulating systemic insulin sensitivity and discuss how PTEN may represent a promising therapeutic strategy for treatment or prevention of T2D.

Artificial Total Disc Replacement Versus Fusion for Lumbar Degenerative Disc Disease: An Update Systematic Review and Meta-Analysis.

AIM: To conduct an updated systematic review and meta-analysis to compare the efficacy and safety between total disc replacement (TDR) and fusion surgery for lumbar degenerative disc disease (LDDD). MATERIAL AND METHODS: We comprehensively searched meta-analyses comparing TDR with fusion through the PubMed, Embase, and Cochrane Library databases. Only randomized controlled trials (RCTs) were selected and collected. The end of the retrieval time was June 2017. Two authors independently extracted the data from the studies after assessing their quality. The statistical software STATA version 12.0 was used to analyze the data. RESULTS: A total of seven RCTs (1706 patients) were included in our analysis. The patients in the TDR group had significantly improved. A greater percentage of these patients were satisfied with the surgery concerning Oswestry disability index, visual analog scale score, and complication rate. In addition, the clinical success in the TDR group was greater than that in the fusion group. Meanwhile, the TDR group had shorter operative time and hospital stay. However, there was no clinical significance regarding blood loss, work status, and reoperation rate between the two groups. CONCLUSION: Our current updated meta-analysis suggests that TDR could be an alternative treatment for LDDD, since it yielded better clinical success and patient satisfaction, shorter hospital stay and operative time, less pain, and lower complication rates than lumbar fusion.

Identification of Cuticular Protein Genes in the Colorado Potato Beetle Leptinotarsa decemlineata (Coleoptera: Chrysomelidae).

Structural cuticular proteins (CPs) are the primary components of insect cuticle, linings of salivary gland, foregut, hindgut and tracheae, and midgut peritrophic membrane. Variation of CPs in insect cuticle can cause penetration resistance to insecticides. Moreover, depletion of specific CP by RNA interference may be a suitable way for the development of potential pest control traits. Leptinotarsa decemlineata (Say) CPs are poorly characterized at present, and therefore, we mined the genome and transcriptome data to better annotate and classify L. decemlineata CPs in this study, by comparison with the annotated CPs of Tribolium castaneum Browse (Coleoptera: Tenebrionidae). We identified 175 CP genes. Except one miscellaneous CP with an 18-amino acid motif, these CPs were classified into 7 families based on motifs and phylogenetic analyses (CPs with a Rebers and Riddiford motif, CPR; CPs analogous to peritrophins, CPAP3 and CPAP1; CPs with a tweedle motif, TWDL; CPs with a 44-amino acid motif, CPF; CPs that are CPF-like, CPFL; and CPs with two to three copies of C-X5-C motif, CPCFC). Leptinotarsa decemlineata CPRs could be categorized into three subfamilies: RR-1 (50), RR-2 (85), and RR-3 (2). The RR-1 proteins had an additional motif with a conserved YTADENGF sequence. The RR-2 members possessed a conserved RDGDVVKG region and three copes of G-x(3)-VV. Few genes were found in TWDL (9), CPAP1 (9), CPAP3 (8), CPF (5), CPFL (4), and CPCFC (2) families. The findings provide valuable information to explore molecular modes of penetration resistance to insecticides and to develop dsRNA-based control method in L. decemlineata.

[Effects of Myospalax baileyi disturbance on plant community at alpine meadow in Three Rivers Headwater Region, China.] / é«˜åŽŸé¼¢é¼ å¹²æ‰°å¯¹ä¸‰æ±ŸæºåŒºé«˜å¯’è‰ç”¸ç¾¤è½ç‰¹å¾çš„å½±å“.

Plateau zokor (Myospalax baileyi) is one of the major rodent species at the alpine meadow in Three Rivers Headwater Region. They eat plant roots, excavate many tunnels and deposit soil on the surface, which result in many exposed mounds to cover the aboveground part of plants. Here, taking plateau zokor mound density in a plot as their disturbance degree, we selected seven plots with different plateau zokor mound densities and one control plot which was not disturbed by plateau zokor to explore the effects of M. baileyi disturbance on community composition, species diversity and productivity. The results showed that, with the increases of available mound density, the dominant species were changed from Cyperaceae and Gramineae species to forb species, including Poly-gonum viviparum, Potentilla anserine, and Polygonum sibiricum. The community coverage and height were significantly decreased. Light or intermediate disturbance by plateau zokor improved species diversity, whereas the evenness index showed no significant change. The changes of community productivity did not support the 'intermediate disturbance hypothesis'. With the increases of plateau zokor disturbance, the aboveground biomass, belowground biomass, total biomass of community significantly decreased. Our findings revealed the effects of plateau zokor's activity on plant community at alpine meadow and presented important information for the management and restoration of degraded grassland and the sustainable utilization of grassland in Three Rivers Headwater Region.

Three-dimensional reconstructions in spine and screw trajectory simulation on 3D digital images: a step by step approach by using Mimics software.

There is a rapidly increasing amount of literature outlining the use of three-dimensional (3D) reconstruction and printing technologies in recent years. However, precise instructive articles which describe step-by-step methods of reconstructing 3D images from computed tomography (CT) or magnetic resonance imaging (MRI) remain limited. To address these issues, this article describes a detailed protocol which will allow the reader to easily perform the 3D reconstruction in their future research, to allow investigation of the appropriate surgical anatomy and allow innovative designs of novel screw fixation techniques or pre-operative surgical planning.

[Allelopathic interactions between invasive plant Solidago canadensis and native plant Phragmites australis].

Taking the seeds of invasive plant Solidago canadensis and native plant Phragmites australis from their mono- and co-dominant communities as allelopathic acceptors, this paper analyzed the differences in the seed germination rate and sprout length after treated with five level (12.5, 25, 50, 100, and 200 mg x mL(-1)) S. canadensis and P. australis extracts, aimed to understand the allelopathic interactions between the two species. The 1000-grain weight and seed germination rate under distilled water treatment of the two species in co-dominated community were greater than those in mono-dominant community. Low level (12.5 and 25 mg x mL(-1)) S. canadensi extracts slightly promoted the seed germination rates of S. canadensis in both mono- and co-dominant communities, but high level (50, 100, and 200 mg x mL(-1)) S. canadensi extracts had strong inhibition effect, especially for the S. canadensis in co-dominated community. No significant patterns were observed about the effects of P. australis extract on S. canadensis seed germination. The sprout length of S. canadensis seeds in both mono- and co-dominant communities decreased with increasing level of S. canadensis extract, but decreased in a fluctuation way with increasing level of P. australis extract. After treated with the extracts of P. australis or S. canadensis, the seed germination rate of P. australis in mono-dominant community was significantly greater than that in co-dominant community (P < 0.05), but there was no significant difference between these two extracts.