ABSTRACT
Harvesting freshwater from fog is one of the possible solutions to the global water scarcity crisis. Surfaces with both hydrophobic and hydrophilic regions are extensively employed for this purpose. Nevertheless, the longevity of these surfaces is still constrained by their delicate surface structures. The hydrophilic zones may become damaged or contaminated after repeated use, thereby compromising their effectiveness in fog collection. The preparation of generally applicable durable superhydrophobic coatings with self-generated Wenzel sites is reported here for long-term efficient and stable fog collection. The coatings are prepared by depositing the poly(tannic acid) coating as the primer layer on various substrates, self-assembly of trichlorovinylsilane into staggered silicone nanofilaments, and then thiol-ene click reaction with 1H,1H,2H,2H-perfluorodecanethiol. The coatings demonstrate remarkable static superhydrophobicity, robust impalement resistance, and stable self-generated Wenzel sites for water droplets. Therefore, the fog collection rate (FCR) of the coatings reaches 2.13 g cm-2 h-1 during 192 h continuous fog collection, which is triple that of bare substrate and outperforms most previous studies. Moreover, the systematic experiments and models have revealed that the key factors for achieving high FCR on superhydrophobic coatings are forming condensed droplets ≈1 mm in critical radius and a Wenzel site proportion of 0.3-0.4.
ABSTRACT
Epileptic encephalopathy (EE) is characterized by seizures that respond poorly to antiseizure drugs, psychomotor delay, and cognitive and behavioral impairments. One of the frequently mutated genes in EE is KCNQ2, which encodes the Kv7.2 subunit of voltage-gated Kv7 potassium channels. Kv7 channels composed of Kv7.2 and Kv7.3 are enriched at the axonal surface, where they potently suppress neuronal excitability. Previously, we reported that the de novo dominant EE mutation M546V in human Kv7.2 blocks calmodulin binding to Kv7.2 and axonal surface expression of Kv7 channels via their intracellular retention. However, whether these pathogenic mechanisms underlie epileptic seizures and behavioral comorbidities remains unknown. Here, we report conditional transgenic cKcnq2+/M547V mice, in which expression of mouse Kv7.2-M547V (equivalent to human Kv7.2-M546V) is induced in forebrain excitatory pyramidal neurons and astrocytes. These mice display early mortality, spontaneous seizures, enhanced seizure susceptibility, memory impairment, and repetitive behaviors. Furthermore, hippocampal pathology shows widespread neurodegeneration and reactive astrocytes. This study demonstrates that the impairment in axonal surface expression of Kv7 channels is associated with epileptic seizures, cognitive and behavioral deficits, and neuronal loss in KCNQ2-related EE.
Subject(s)
Epileptic Syndromes/genetics , KCNQ2 Potassium Channel/genetics , Nerve Tissue Proteins/genetics , Animals , Behavior, Animal , Cognitive Dysfunction , Epileptic Syndromes/pathology , Epileptic Syndromes/psychology , Female , Gliosis , Hippocampus/pathology , KCNQ2 Potassium Channel/metabolism , Kainic Acid , Male , Mice, Inbred C57BL , Mice, Transgenic , Nerve Tissue Proteins/metabolism , Pyramidal Cells/metabolismABSTRACT
RESEARCH QUESTION: What is the effect of miR-122 on the progression and recovery of fibrosis in Asherman's syndrome? DESIGN: Endometrial tissue was collected from 21 patients, 11 with intrauterine adhesion (IUA) and 10 without IUA. Quantitative real-time polymerase chain reaction, immunofluorescence and Western blot were applied to observe the expression of mRNAs/miRNAs and protein, respectively. The endometrial physical injury was carried out in C57BL/6 mice to create an endometrial fibrosis model, with intrauterine injection of adenovirus to compare the antifibrosis and repair function of miR-122 on endometrium. The morphology of the uterus was observed using haematoxylin and eosin staining, and fibrosis markers were detected by immunohistochemistry. RESULTS: miR-122 expression was reduced in patients with IUAs, accompanied by fibrosis. MiR-122 overexpression reduced the degree of fibrosis in endometrial stromal cells. Further molecular analyses demonstrated that miR-122 inhibited fibrosis through the TGF-ß/SMAD pathway by directly targeting the 3' untranslated region of SMAD family member 3, suppressing its expression. Notably, miR-122 promoted endometrial regeneration and recovery of pregnancy capacity in a mouse endometrial injury model. CONCLUSIONS: miR-122 is a critical regulator for repair of endometrial fibrosis and provided new insight for the clinical treatment of intrauterine adhesions.
Subject(s)
Gynatresia , MicroRNAs , Uterine Diseases , Mice , Animals , Female , Pregnancy , Humans , Transforming Growth Factor beta/metabolism , Mice, Inbred C57BL , Uterine Diseases/genetics , Uterine Diseases/pathology , Endometrium/metabolism , MicroRNAs/genetics , MicroRNAs/metabolism , Tissue Adhesions , Disease Models, Animal , FibrosisABSTRACT
Accessory spleen (AS) refers to single or multiple splenic tissues which appear outside the relative normal spleen position results from embryonic dysplasia similar in structure and function to the spleen. AS is frequently observed in the splenic hilus and or adjacent to the tail of pancreas, and only a few cases occurred in the pelvic cavity. We present an extremely rare AS case in urachus, which was initially considered as an urachal neoplasm revealed on CT images with big mass. However, the postoperative pathology confirmed it was an AS that had not been reported at urachal before. Urachal AS can be misdiagnosed as a tumor, so it is vital to make an accurate imaging preoperative diagnosis to avoid unnecessary biopsy and surgery.
Subject(s)
Urachus , Humans , Urachus/diagnostic imaging , Urachus/surgery , Spleen/diagnostic imaging , Spleen/surgery , Tomography, X-Ray Computed , BiopsyABSTRACT
OBJECTIVE: STriatal-Enriched protein tyrosine Phosphatase (STEP) is a brain-specific tyrosine phosphatase. Membrane-bound STEP61 is the only isoform expressed in hippocampus and cortex. Genetic deletion of STEP enhances excitatory synaptic currents and long-term potentiation in the hippocampus. However, whether STEP61 affects seizure susceptibility is unclear. Here we investigated the effects of STEP inhibitor TC-2153 on seizure propensity in a murine model displaying kainic acid (KA)-induced status epilepticus and its effect on hippocampal excitability. METHODS: Adult male and female C57BL/6J mice received intraperitoneal injection of either vehicle (2.8% dimethylsulfoxide [DMSO] in saline) or TC-2153 (10 mg/kg) and then either saline or KA (30 mg/kg) 3 h later before being monitored for behavioral seizures. A subset of female mice was ovariectomized (OVX). Acute hippocampal slices from Thy1-GCaMP6s mice were treated with either DMSO or TC-2153 (10 µM) for 1 h, and then incubated in artificial cerebrospinal fluid (ACSF) and potassium chloride (15 mM) for 2 min prior to live calcium imaging. Pyramidal neurons in dissociated rat hippocampal culture (DIV 8-10) were pre-treated with DMSO or TC-2153 (10 µM) for 1 h before whole-cell patch-clamp recording. RESULTS: TC-2153 treatment significantly reduced KA-induced seizure severity, with greater trend seen in female mice. OVX abolished this TC-2153-induced decrease in seizure severity in female mice. TC-2153 application significantly decreased overall excitability of acute hippocampal slices from both sexes. Surprisingly, TC-2153 treatment hyperpolarized resting membrane potential and decreased firing rate, sag voltage, and hyperpolarization-induced current (Ih ) of cultured hippocampal pyramidal neurons. SIGNIFICANCE: This study is the first to demonstrate that pharmacological inhibition of STEP with TC-2153 decreases seizure severity and hippocampal activity in both sexes, and dampens hippocampal neuronal excitability and Ih . We propose that the antiseizure effects of TC-2153 are mediated by its unexpected action on suppressing neuronal intrinsic excitability.
Subject(s)
Dimethyl Sulfoxide , Hippocampus , Animals , Benzothiepins , Dimethyl Sulfoxide/adverse effects , Dimethyl Sulfoxide/metabolism , Female , Kainic Acid/pharmacology , Male , Mice , Mice, Inbred C57BL , Rats , Seizures/chemically induced , Seizures/metabolismABSTRACT
BACKGROUND: The amino acid sequence of proteins generally carries all the necessary information for acquisition of native conformations, but the vectorial nature of translation can additionally determine the folding outcome. Such consideration is particularly relevant in human diseases associated to inherited mutations leading to structural instability, aggregation, and degradation. Mutations in the KCNQ2 gene associated with human epilepsy have been suggested to cause misfolding of the encoded Kv7.2 channel. Although the effect on folding of mutations in some domains has been studied, little is known of the way pathogenic variants located in the calcium responsive domain (CRD) affect folding. Here, we explore how a Kv7.2 mutation (W344R) located in helix A of the CRD and associated with hereditary epilepsy interferes with channel function. RESULTS: We report that the epilepsy W344R mutation within the IQ motif of CRD decreases channel function, but contrary to other mutations at this site, it does not impair the interaction with Calmodulin (CaM) in vitro, as monitored by multiple in vitro binding assays. We find negligible impact of the mutation on the structure of the complex by molecular dynamic computations. In silico studies revealed two orientations of the side chain, which are differentially populated by WT and W344R variants. Binding to CaM is impaired when the mutated protein is produced in cellulo but not in vitro, suggesting that this mutation impedes proper folding during translation within the cell by forcing the nascent chain to follow a folding route that leads to a non-native configuration, and thereby generating non-functional ion channels that fail to traffic to proper neuronal compartments. CONCLUSIONS: Our data suggest that the key pathogenic mechanism of Kv7.2 W344R mutation involves the failure to adopt a configuration that can be recognized by CaM in vivo but not in vitro.
Subject(s)
Epilepsy , KCNQ2 Potassium Channel/genetics , Amino Acid Sequence , Calcium/metabolism , Calmodulin/genetics , Calmodulin/metabolism , Epilepsy/genetics , Humans , KCNQ2 Potassium Channel/metabolism , MutationABSTRACT
Neuronal Kv7/KCNQ channels are voltage-gated potassium channels composed of Kv7.2/KCNQ2 and Kv7.3/KCNQ3 subunits. Enriched at the axonal membrane, they potently suppress neuronal excitability. De novo and inherited dominant mutations in Kv7.2 cause early onset epileptic encephalopathy characterized by drug resistant seizures and profound psychomotor delay. However, their precise pathogenic mechanisms remain elusive. Here, we investigated selected epileptic encephalopathy causing mutations in calmodulin (CaM)-binding helices A and B of Kv7.2. We discovered that R333W, K526N, and R532W mutations located peripheral to CaM contact sites decreased axonal surface expression of heteromeric channels although only R333W mutation reduced CaM binding to Kv7.2. These mutations also altered gating modulation by phosphatidylinositol 4,5-bisphosphate (PIP2), revealing novel PIP2 binding residues. While these mutations disrupted Kv7 function to suppress excitability, hyperexcitability was observed in neurons expressing Kv7.2-R532W that displayed severe impairment in voltage-dependent activation. The M518â¯V mutation at the CaM contact site in helix B caused most defects in Kv7 channels by severely reducing their CaM binding, K+ currents, and axonal surface expression. Interestingly, the M518â¯V mutation induced ubiquitination and accelerated proteasome-dependent degradation of Kv7.2, whereas the presence of Kv7.3 blocked this degradation. Furthermore, expression of Kv7.2-M518V increased neuronal death. Together, our results demonstrate that epileptic encephalopathy mutations in helices A and B of Kv7.2 cause abnormal Kv7 expression and function by disrupting Kv7.2 binding to CaM and/or modulation by PIP2. We propose that such multiple Kv7 channel defects could exert more severe impacts on neuronal excitability and health, and thus serve as pathogenic mechanisms underlying Kcnq2 epileptic encephalopathy.
Subject(s)
Axons/metabolism , Brain Diseases/metabolism , Epilepsy, Generalized/metabolism , KCNQ2 Potassium Channel/biosynthesis , Neurons/metabolism , Phosphatidylinositols/biosynthesis , Amino Acid Sequence , Animals , Axons/pathology , Brain Diseases/genetics , Brain Diseases/pathology , Epilepsy, Generalized/genetics , Epilepsy, Generalized/pathology , Gene Expression , HEK293 Cells , Humans , KCNQ2 Potassium Channel/chemistry , KCNQ2 Potassium Channel/genetics , Neurons/pathology , Phosphatidylinositols/genetics , Protein Structure, Secondary , RatsABSTRACT
Disease problems cause major economic losses for the aquaculture industries. In catfish, enteric septicemia of catfish (ESC), caused by the bacterial pathogen Edwardsiella ictaluri, is the leading disease problem, causing tens of millions of dollars of annual economic losses. In this study, we conducted a genome-wide association study to determine quantitative trait loci (QTL) for resistance against ESC using an interspecific hybrid system. Five hundred fish were used in the analysis and 192 phenotypic extremes were used for genotyping with the catfish 250K SNP arrays. A genomic region on linkage group (LG) 1 was found significantly associated with ESC disease resistance. In addition, two suggestively associated QTL for ESC resistance were identified on LG 12 and LG 16. The nlrc3 duplicates were identified within all the three QTL, suggesting their importance in association with the QTL. Within the significant QTL on LG 1, 16 genes with known functions in immunity were identified. Of particular interest is the nck1 gene nearby the most significantly associated SNP. Nck1 was known to function as an adaptor to facilitating the pathogenesis of enteropathogenic Escherichia coli (EPEC) in humans. E. ictaluri and EPEC pathogens belong to the same bacterial family and share many common characteristics. The fact that nck1 is mapped in the QTL and that it was significantly upregulated in channel catfish intestine after ESC challenge suggested its candidacy of being involved in resistance/susceptibility of ESC.
Subject(s)
Catfishes , Edwardsiella ictaluri/physiology , Enterobacteriaceae Infections/veterinary , Fish Diseases/genetics , Sepsis/veterinary , Animals , Crosses, Genetic , Disease Resistance , Enterobacteriaceae Infections/genetics , Enterobacteriaceae Infections/immunology , Enterobacteriaceae Infections/microbiology , Fish Diseases/immunology , Fish Diseases/microbiology , Genome-Wide Association Study , Quantitative Trait Loci , Sepsis/genetics , Sepsis/immunologyABSTRACT
Heat shock proteins 70/110 (Hsp70/110) are a family of conserved ubiquitously expressed heat shock proteins which are produced by cells in response to exposure to stressful conditions. Besides the chaperone and housekeeping functions, they are also known to be involved in immune response during infection. In this study, we identified 16 Hsp70/110 geness in channel catfish (Ictalurus punctatus) through in silico analysis using RNA-Seq and genome databases. Among them 12 members of Hsp70 (Hspa) family and 4 members of Hsp110 (Hsph) family were identified. Phylogenetic and syntenic analyses provided strong evidence in supporting the orthologies of these HSPs. In addition, we also determined the expression patterns of Hsp70/110 genes after Flavobacterium columnare and Edwardsiella ictaluri infections by meta-analyses, for the first time in channel catfish. Ten out of sixteen genes were significantly up/down-regulated after bacterial challenges. Specifically, nine genes were found significantly expressed in gill after F. columnare infection. Two genes were found significantly expressed in intestine after E. ictaluri infection. Pathogen-specific pattern and tissue-specific pattern were found in the two infections. The significantly regulated expressions of catfish Hsp70 genes after bacterial infections suggested their involvement in immune response in catfish.
Subject(s)
Enterobacteriaceae Infections/veterinary , Fish Diseases/genetics , Fish Proteins/genetics , Flavobacteriaceae Infections/veterinary , Gene Expression Regulation/immunology , HSP70 Heat-Shock Proteins/genetics , Ictaluridae/genetics , Animals , Edwardsiella ictaluri/physiology , Enterobacteriaceae Infections/genetics , Enterobacteriaceae Infections/immunology , Enterobacteriaceae Infections/microbiology , Fish Diseases/immunology , Fish Diseases/microbiology , Fish Proteins/metabolism , Flavobacteriaceae Infections/genetics , Flavobacteriaceae Infections/immunology , Flavobacteriaceae Infections/microbiology , Flavobacterium/physiology , Genome , HSP70 Heat-Shock Proteins/metabolism , Ictaluridae/metabolism , Molecular Sequence Data , Phylogeny , Sequence Analysis, DNA , SyntenyABSTRACT
OBJECTIVE: To identify the prevalence of unexpected uterine sarcoma after total laparoscopic or abdominal hysterectomy for presumed leiomyoma and compare clinical consequences after hysterectomy with and without transvaginal scalpel morcellation (TVSM). METHODS: In this retrospective study, the medical records of patients who had unexpected uterine sarcoma after total laparoscopic or abdominal hysterectomy for presumed leiomyoma between 2009 and 2013 were reviewed. RESULTS: Among 3021 patients who underwent total hysterectomy for presumed leiomyoma, 18 (1/168, 0.60%) had unexpected uterine sarcoma (5 [1/604, 0.17%] had leiomyosarcoma and 13 [1/232, 0.43%] had low-grade endometrial stromal sarcoma). The risk of unexpected leiomyosarcoma increased steadily in ages from the 40s to the 50s, whereas the risk of unexpected endometrial stromal sarcoma (ESS) decreased steadily in the same period. The unexpected sarcoma was identified in 7 (1/158, 0.63%) of 1104 patients treated by laparoscopy and 11 (1/174, 0.57%) of 1917 patients by laparotomy. Transvaginal scalpel morcellation was performed to extract the uterus in majority (78.53%) of the patients with total laparoscopic hysterectomy. Sixteen (88.89%) cases were low grade, and 2 (11.11%) were high grade: 17 at stage I and 1 at stage II. Nine patients underwent a secondary operation, and 11 patients received adjuvant therapy postoperatively. Except for 1 patient with additional power morcellation, all patients with unexpected ESS survived without recurrence after total hysterectomy with and without TVSM, with mean follow-ups of 25.20 (16-36) months and 32.57 (21-50) months, respectively. CONCLUSIONS: The overall incidence of unexpected uterine sarcoma after total hysterectomy for presumed leiomyoma was low. Low-grade endometrial stromal sarcoma was the dominant subtype of unexpected uterine sarcoma in the present study. Currently, incidental TVSM of unexpected ESS during total laparoscopic hysterectomy seemed to cause no additional increase in sarcoma dissemination in the short-term.
Subject(s)
Hysterectomy/adverse effects , Leiomyoma/surgery , Morcellation/methods , Sarcoma/pathology , Uterine Neoplasms/surgery , Vagina/surgery , Adult , Aged , Female , Follow-Up Studies , Humans , Laparoscopy , Leiomyoma/complications , Leiomyoma/pathology , Middle Aged , Neoplasm Staging , Prognosis , Retrospective Studies , Sarcoma/etiology , Survival Rate , Uterine Neoplasms/complications , Uterine Neoplasms/pathologyABSTRACT
BACKGROUND: Cytochrome P450s (CYPs) encode one of the most diverse enzyme superfamily in nature. They catalyze oxidative reactions of endogenous molecules and exogenous chemicals. METHODS: We identified CYPs genes through in silico analysis using EST, RNA-Seq and genome databases of channel catfish. Phylogenetic analyses and conserved syntenic analyses were conducted to determine their identities and orthologies. Meta-analysis of RNA-Seq databases was conducted to analyze expression profile of CYP genes following bacterial infection. RESULTS: A full set of 61 CYP genes was identified and characterized in channel catfish. Phylogenetic tree and conserved synteny provided strong evidence of their identities and orthorlogy. Lineage-specific gene duplication was evident in a number of clans in channel catfish. CYP46A1 is missing in the catfish genome as observed with syntenic analysis and RT-PCR analysis. Thirty CYPs were found up- or down-regulated in liver, while seven and eight CYPs were observed regulated in intestine and gill following bacterial infection. CONCLUSION: We systematically identified and characterized a full set of 61 CYP genes in channel catfish and studied their expression profiles after bacterial infection. While bacterial challenge altered the expression of large numbers of CYP genes, the mechanisms and significance of these changes are not known. GENERAL SIGNIFICANCE: This work provides an example to systematically study CYP genes in non-model species. Moreover, it provides a basis for further toxicological and physiological studies in channel catfish.
Subject(s)
Cytochrome P-450 Enzyme System , Fish Proteins , Gene Expression Regulation, Enzymologic/physiology , Genome/physiology , Ictaluridae , Phylogeny , Animals , Cytochrome P-450 Enzyme System/biosynthesis , Cytochrome P-450 Enzyme System/genetics , Fish Proteins/biosynthesis , Fish Proteins/genetics , Ictaluridae/genetics , Ictaluridae/metabolismABSTRACT
BACKGROUND: Columnaris causes severe mortalities among many different wild and cultured freshwater fish species, but understanding of host resistance is lacking. Catfish, the primary aquaculture species in the United States, serves as a great model for the analysis of host resistance against columnaris disease. Channel catfish in general is highly resistant to the disease while blue catfish is highly susceptible. F2 generation of hybrids can be produced where phenotypes and genotypes are segregating, providing a useful system for QTL analysis. To identify genes associated with columnaris resistance, we performed a genome-wide association study (GWAS) using the catfish 250 K SNP array with 340 backcross progenies derived from crossing female channel catfish (Ictalurus punctatus) with male F1 hybrid catfish (female channel catfish I. punctatus × male blue catfish I. furcatus). RESULTS: A genomic region on linkage group 7 was found to be significantly associated with columnaris resistance. Within this region, five have known functions in immunity, including pik3r3b, cyld-like, adcyap1r1, adcyap1r1-like, and mast2. In addition, 3 additional suggestively associated QTL regions were identified on linkage groups 7, 12, and 14. The resistant genotypes on the QTLs of linkage groups 7 and 12 were found to be homozygous with both alleles being derived from channel catfish. The paralogs of the candidate genes in the suggestively associated QTL of linkage group 12 were found on the QTLs of linkage group 7. Many candidate genes on the four associated regions are involved in PI3K pathway that is known to be required by many bacteria for efficient entry into the host. CONCLUSION: The GWAS revealed four QTLs associated with columnaris resistance in catfish. Strikingly, the candidate genes may be arranged as functional hubs; the candidate genes within the associated QTLs on linkage groups 7 and 12 are not only co-localized, but also functionally related, with many of them being involved in the PI3K signal transduction pathway, suggesting its importance for columnaris resistance.
Subject(s)
Catfishes/genetics , Disease Resistance/genetics , Genome-Wide Association Study , Quantitative Trait Loci , Animals , Female , Genetic Association Studies , Genetic Linkage , Genomics , Linkage Disequilibrium , Male , Mortality , Polymorphism, Single NucleotideABSTRACT
Interfacial solar evaporation (ISE) holds considerable promise to solve fresh water shortage, but it is challenging to achieve high evaporation rate (Reva) and fresh water yield in close system. Here, we report design and preparation of MOF-based solar evaporators with hierarchical microporous/nanobridged/nanogranular structures for rapid ISE and fresh water collection in close system. The evaporators are fabricated by growing silicone nanofilaments with variable length as nanobridges on a microporous silicone sponge followed by grafting with polydopamine nanoparticles and Cu-MOF nanocrystals. Integration of the unique structure and excellent photothermal composites endows the evaporators with high Reva of 3.5-20 wt% brines (3.60-2.90 kg m-2 h-1 in open system and 2.38-1.44 kg m-2 h-1 in close system) under simulated 1 sun, high Reva under natural sunlight, excellent salt resistance and high fresh water yield, which surpass most state-of-the-art evaporators. Moreover, when combined with a superhydrophilic cover, the evaporators show much higher average Reva of real seawater, remarkable fresh water yield and excellent long-term stability over one month continuous ISE under natural sunlight. The findings here will promote the development of advanced evaporators via microstructure engineering and their real-world ISE applications.
ABSTRACT
Solitary fibrous tumor (SFT) is a rare spindle cell tumor originating from mesenchymal tissue, and even rarer when it occurs intracranially. This case report described a 42-year-old man who presented with headache and limb weakness for more than 10 days. Magnetic resonance imaging (MRI) showed a well-defined multicompartmental cystic space-occupying lesion in the left occipital region, with surrounding edema and a compressed left lateral ventricle, the mass growing across the cerebellar vermis, which was initially diagnosed as hemangioblastoma. Neurosurgery was utilized to successfully remove the mass, and intracranial solitary fibrous tumor (ISFT) was identified by postoperative pathological analysis. Here, this article describes the imaging manifestations and pathologic features of a case of cystic intracranial solitary fibrous tumor, aiming to improve the understanding and diagnosis of this disease in order to provide an accurate therapy plan.
ABSTRACT
Circulating tumor cells (CTCs) captured from the bloodstream of patients with solid tumors have the potential to accelerate precision oncology by providing insight into tumor biology, disease progression and response to treatment. However, their potential is hampered by the lack of standardized CTC enrichment platforms across tumor types. EpCAM-based CTC enrichment, the most commonly used platform, is limited by EpCAM downregulation during metastasis and the low EpCAM expression in certain tumor types, including the highly prevalent and lethal NSCLC. In this study we demonstrate that Transferrin Receptor (TfR) is a selective, efficient biomarker for CTC identification and capture in patients with prostate, pancreatic and NSCLC. TfR identifies significantly higher CTC counts than EpCAM, and TfR + -CTC enumeration correlates with disease progression in metastatic prostate and pancreatic cancers, and overall survival and osimetrinib-resistance in non-small cell lung cancer (NSCLC). Profiling of TfR + -CTCs provides a snapshot of the molecular landscape of each respective tumor type and identifies potential mechanisms underlying treatment response to EGFR TKi and immune checkpoint inhibitors in NSCLC. One sentence summary: Transferrin Receptor identifies circulating tumor cells in solid tumors.
ABSTRACT
Temperature is one of the most prominent abiotic factors affecting ectotherms. Most fish species, as ectotherms, have extraordinary ability to deal with a wide range of temperature changes. While the molecular mechanism underlying temperature adaptation has long been of interest, it is still largely unexplored with fish. Understanding of the fundamental mechanisms conferring tolerance to temperature fluctuations is a topic of increasing interest as temperature may continue to rise as a result of global climate change. Catfish have a wide natural habitat and possess great plasticity in dealing with environmental variations in temperature. However, no studies have been conducted at the transcriptomic level to determine heat stress-induced gene expression. In the present study, we conducted an RNA-Seq analysis to identify heat stress-induced genes in catfish at the transcriptome level. Expression analysis identified a total of 2,260 differentially expressed genes with a cutoff of twofold change. qRT-PCR validation suggested the high reliability of the RNA-Seq results. Gene ontology, enrichment, and pathway analyses were conducted to gain insight into physiological and gene pathways. Specifically, genes involved in oxygen transport, protein folding and degradation, and metabolic process were highly induced, while general protein synthesis was dramatically repressed in response to the lethal temperature stress. This is the first RNA-Seq-based expression study in catfish in response to heat stress. The candidate genes identified should be valuable for further targeted studies on heat tolerance, thereby assisting the development of heat-tolerant catfish lines for aquaculture.
Subject(s)
Catfishes/genetics , Gene Expression Profiling , Heat-Shock Response/genetics , Oxygen/metabolism , Protein Biosynthesis/genetics , Protein Folding , Proteolysis , Sequence Analysis, RNA , Adaptation, Physiological/genetics , Animals , Biological Transport/genetics , Body Size/genetics , Catfishes/anatomy & histology , Gills/metabolism , Liver/metabolism , Molecular Sequence Annotation , Reproducibility of Results , Reverse Transcriptase Polymerase Chain Reaction , Signal Transduction/genetics , Transcriptome/geneticsABSTRACT
BACKGROUND: The application of RNA-seq has accelerated gene expression profiling and identification of gene-associated SNPs in many species. However, the integrated studies of gene expression along with SNP mapping have been lacking. Coupling of RNA-seq with bulked segregant analysis (BSA) should allow correlation of expression patterns and associated SNPs with the phenotypes. RESULTS: In this study, we demonstrated the use of bulked segregant RNA-seq (BSR-Seq) for the analysis of differentially expressed genes and associated SNPs with disease resistance against enteric septicemia of catfish (ESC). A total of 1,255 differentially expressed genes were found between resistant and susceptible fish. In addition, 56,419 SNPs residing on 4,304 unique genes were identified as significant SNPs between susceptible and resistant fish. Detailed analysis of these significant SNPs allowed differentiation of significant SNPs caused by genetic segregation and those caused by allele-specific expression. Mapping of the significant SNPs, along with analysis of differentially expressed genes, allowed identification of candidate genes underlining disease resistance against ESC disease. CONCLUSIONS: This study demonstrated the use of BSR-Seq for the identification of genes involved in disease resistance against ESC through expression profiling and mapping of significantly associated SNPs. BSR-Seq is applicable to analysis of genes underlining various performance and production traits without significant investment in the development of large genotyping platforms such as SNP arrays.
Subject(s)
Catfishes/genetics , Disease Resistance/genetics , Enterobacteriaceae Infections/veterinary , Fish Diseases/genetics , Sepsis/genetics , Animals , Edwardsiella ictaluri , Enterobacteriaceae Infections/genetics , Fish Diseases/microbiology , Gene Expression Profiling , Polymorphism, Single Nucleotide , Sepsis/microbiology , Sepsis/veterinary , Sequence Analysis, RNAABSTRACT
A full-length sequence encoding the estrogen receptor beta was isolated from half-smooth tongue sole, Cynoglossus semilaevis (hstsERß) using reverse transcription-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends procedures. The hstsERß cDNA clone was found to contain 1,791 nucleotides including an open reading frame that encodes 578 amino acids. The deduced hstsERß protein consisted of six nuclear receptor-characteristic domains. Based on a phylogenetic analysis, the hstsERß C and E domains are highly conserved compared to other fishes. The potential phosphorylation sites for PKC, CK-2 and PTK are also found in this protein. Highest amino acid identities were found for hstsERß with common carp (Cyprinus carpio) ERß (76 %) and Japanese flounder (Paralichthys olivaceus) ERß (76 %). Tissue expression analysis confirmed that the hstsERß was widely distributed and predominantly expressed in testis, brain and liver. Seasonal changes in the testis, brain and liver expression profiles of hstsERß were examined by RT-PCR; the present results suggest that level of hstsERß in brain increased to the highest then decreases with gonadal growth; whereas in the testis and liver, the hstsERß mRNA level dropped to lowest then slightly increased.
Subject(s)
Estrogen Receptor beta/genetics , Estrogen Receptor beta/metabolism , Flatfishes , Phylogeny , Amino Acid Sequence , Analysis of Variance , Animals , Base Sequence , Brain/metabolism , Cloning, Molecular , Cluster Analysis , Conserved Sequence/genetics , DNA Primers/genetics , DNA, Complementary/genetics , Estrogen Receptor beta/chemistry , Gene Expression Profiling , Liver/metabolism , Male , Molecular Sequence Data , Nucleic Acid Amplification Techniques , Open Reading Frames/genetics , Protein Structure, Tertiary , Radioimmunoassay , Reverse Transcriptase Polymerase Chain Reaction , Sequence Analysis, DNA , Sequence Homology , Testis/metabolismABSTRACT
Mucinous tubular and spindle cell carcinoma of the kidney (MTSCC) is a rare subtype of renal cancer. It consists of tubules separated by mucus stroma and a spindle cell. Few cases have been reported; thus, the imaging features of MTSCC are not well characterized. An MTSCC in the left kidney of a 65-year-old woman was incidentally discovered during a medical checkup. A review of the patient's medical history revealed that this kidney lump had an indolent growth process. The current study presented this case and reviewed the pathological features, imaging findings and treatment options of MTSCC to strengthen the recognition of this rare renal neoplasm by radiologists.
ABSTRACT
BACKGROUND: Upon the completion of whole genome sequencing, thorough genome annotation that associates genome sequences with biological meanings is essential. Genome annotation depends on the availability of transcript information as well as orthology information. In teleost fish, genome annotation is seriously hindered by genome duplication. Because of gene duplications, one cannot establish orthologies simply by homology comparisons. Rather intense phylogenetic analysis or structural analysis of orthologies is required for the identification of genes. To conduct phylogenetic analysis and orthology analysis, full-length transcripts are essential. Generation of large numbers of full-length transcripts using traditional transcript sequencing is very difficult and extremely costly. RESULTS: In this work, we took advantage of a doubled haploid catfish, which has two sets of identical chromosomes and in theory there should be no allelic variations. As such, transcript sequences generated from next-generation sequencing can be favorably assembled into full-length transcripts. Deep sequencing of the doubled haploid channel catfish transcriptome was performed using Illumina HiSeq 2000 platform, yielding over 300 million high-quality trimmed reads totaling 27 Gbp. Assembly of these reads generated 370,798 non-redundant transcript-derived contigs. Functional annotation of the assembly allowed identification of 25,144 unique protein-encoding genes. A total of 2,659 unique genes were identified as putative duplicated genes in the catfish genome because the assembly of the corresponding transcripts harbored PSVs or MSVs (in the form of pseudo-SNPs in the assembly). Of the 25,144 contigs with unique protein hits, around 20,000 contigs matched 50% length of reference proteins, and over 14,000 transcripts were identified as full-length with complete open reading frames. The characterization of consensus sequences surrounding start codon and the stop codon confirmed the correct assembly of the full-length transcripts. CONCLUSIONS: The large set of transcripts assembled in this study is the most comprehensive set of genome resources ever developed from catfish, which will provide the much needed resources for functional genome research in catfish, serving as a reference transcriptome for genome annotation, analysis of gene duplication, gene family structures, and digital gene expression analysis. The putative set of duplicated genes provide a starting point for genome scale analysis of gene duplication in the catfish genome, and should be a valuable resource for comparative genome analysis, genome evolution, and genome function studies.