Search

The phagosome-lysosome (PL) signaling pathway plays an important role in the immune system of fish. However, the molecular mechanism underlying PL signaling in fish against Vibrio vulnificus infection remains still unclear. In this study, the half-smooth tongue sole (Cynoglossus semilaevis) infected with V. vulnificus strain ST-6 was used as an experiment model, the immunohistochemical features were evaluated, and spatiotemporal expression analysis of genes and proteins related to PL pathway was verified, respectively. The results showed that the positive staining was observed in the various tissues such as liver, intestine, kidney, spleen, and gills of the infected fish. The differential expressions of PL-related proteins were screened and identified by iTRAQ analysis, and their gene expressions were determined by qRT-PCR detection. The spatiotemporal expression patters were confirmed and most of PL-related genes tended to be downregulated during the early infection at 12 and 36 h post-infection (hpi). Moreover, the protein-protein interaction (PPI) analysis was described for PL pathway. This is the first report on the PL response of C. semilaevis to V. vulnificus infection, which will provide a scientific reference for further elucidating the molecular mechanism of the phagosome-lysosome in marine fish. [ABSTRACT FROM AUTHOR]

Vibrio spp. are major pathogens responsible for mortality and disease in various marine aquaculture organisms. Effective disease control and genetic breeding strategies rely heavily on understanding host vibriosis resistance mechanisms. The Chinese tongue sole (Cynoglossus semilaevis) is economically vital but suffers from substantial mortalities due to vibriosis. Through continuous selective breeding, we have successfully obtained vibriosis-resistant families of this species. In this study, we conducted RNA-seq analysis on three organs, including liver, spleen and intestine from selected resistant and susceptible tongue soles. Additionally, we integrated these data with our previously published RNA-seq datasets of skin and gill, enabling the construction of organ-specific transcriptional profiles and a comprehensive gene co-expression network elucidating the differences in vibriosis resistance. Furthermore, we identified 12 modules with organ-specific functional implications. Overall, our findings provide a valuable resource for investigating the molecular basis of vibriosis resistance in fish, offering insights into target genes and pathways essential for molecular selection and genetic manipulation to enhance vibriosis resistance in fish breeding programs. [ABSTRACT FROM AUTHOR]

The assembly of W and Y chromosomes poses significant challenges in vertebrate genome sequencing and assembly. Here, we successfully assembled the W chromosome of Verasper variegatus with a length of 20.48 Mb by combining population and PacBio HiFi sequencing data. It was identified as a young sex chromosome and showed signs of expansion in repetitive sequences. The major component of the expansion was Ty3/Gypsy. The ancestral Osteichthyes karyotype consists of 24 protochromosomes. The sex chromosomes in four Pleuronectiformes species derived from a pair of homologous protochromosomes resulting from a whole-genome duplication event in teleost fish, yet with different sex-determination systems. V. variegatus and Cynoglossus semilaevis adhere to the ZZ/ZW system, while Hippoglossus stenolepis and H. hippoglossus follow the XX/XY system. Interestingly, V. variegatus and H. hippoglossus derived from one protochromosome, while C. semilaevis and H. stenolepis derived from another protochromosome. Our study provides valuable insights into the evolution of sex chromosomes in flatfish and sheds light on the important role of whole-genome duplication in shaping the evolution of sex chromosomes. [ABSTRACT FROM AUTHOR]

The purpose of this study was to investigate the microplastic (MP) pollution in sediments, surface waters, and four fish species in the northern coast of the Persian Gulf. Sampling was conducted in seven important regions during December 2019. The abundance of MPs was respectively 190 ± 35.5 items/kg dry weight for sediments, 9.28 ± 2.1 items/km2 for surface waters, and 0.33 ± 0.05 items/individual for fish. There was no correlation between MP abundance in surface water, sediment, and fish samples. Except for Cynoglossus arel, abundance of isolated MPs did not show significant relationships with body weight, body length, and gastrointestinal tract weight (P > 0.05). MPs were ranged from 0.3 to 5 mm in size and were prevailed by fiber in shape; black, red, and blue in color; and polypropylene and polyethylene in polymer. This study may help in increasing our knowledge regarding MP pollution in marine water systems and biota. [ABSTRACT FROM AUTHOR]

The transient receptor potential vanilloid 4 (TRPV4), another Ca2+ entry channel, belongs to the vanilloid subfamily and responds to a number of different physical and chemical stimuli and exists widely in mammals. However, our understanding of the TRPV4 in fish remains poor. Therefore, we studied the TRPV4 gene from Cynoglossus semilaevis, named CsTRPV4 that encodes a putative protein of 870 amino acids common in structure and characteristic of mammalian TRPV4, including the domains of ANK repeats, six TM, TRP domain, and CaMBD. The CsTRPV4 was expressed ubiquitously in examined tissues: higher expression in the heart, spleen, testis, and eye, but lower expression in kidney and liver. Surprisingly, the expression of CsTRPV4 in lateral line was significantly higher than in many other tissues as the CsTRPV4 was expressed significantly in the free neuromasts. In addition, CsTRPV4 in the free neuromast from the larval fish was significantly expressed in the hair cells of the free neuromasts. Therefore, the free neuromasts can act as a mechano-sensor to the mechanical stimulation in molecular level in C. semilaevis, which lays a foundation for further study of the functions of the free neuromasts. [ABSTRACT FROM AUTHOR]

At present, the function and profile of long noncoding RNAs (IncRNAs) in half-smooth tongue soles Cynoglossus semilaevis remain poorly understood. Lherefore, we identified IncRNAs in the species using large-scale deep sequencing approaches. A total of 96 726 222 clean reads and 2497 IncRNAs were obtained from the compound samples. In total, 1 694 known IncRNAs and 803 novel IncRNAs were identified. Most of the novel IncRNAs distributed mainly in a range of 200–3 000 nt and contained 2–6 exons. Six novel IncRNAs were selected for expression analysis by quantitative reverse transcription-polymerase chain reaction, of which lnc_770 and lnc_150 were expressed mainly in the ovary. Lhis study provides a valuable resource for IncRNA studies of half-smooth tongue sole and improves our understanding of the function of non-coding RNA in fish. [ABSTRACT FROM AUTHOR]

Cynoglossus yokomaru sp. nov., previously known as Cynoglossus sp., called "Otafuku-genko" (the standard Japanese name), is described based on 15 specimens [64–226 mm in standard length (SL)] collected at depths of 73–157 m in the East China Sea and Yellow Sea. The new species belongs to the Cynoglossus kopsii complex as redefined in this paper and clearly differs from its congeners by the following combination of characters: two ocular-side lateral lines (dorsolateral and midlateral lines present, ventrolateral line absent); no blind-side lateral line; ctenoid scales present on both sides; two ocular-side nostrils; ocular-side posterior nostril immediately anterior to a point between eyes; head length (HL) shorter than its depth; posterior margin of upper jaw on ocular side not reaching below posterior margin of lower eye; body depth 25.8–30.2% SL; upper supraorbital length 35.5–47.3% HL; lower supraorbital length 48.8–64.3% HL; horizontal diameter of lower eye 10.1–17.4% HL; a long and continuous dorsolateral line with 61–70 scales; midlateral-line scales 4–5 + 65–68 = 69–73; scales above dorsolateral line 3–4; scales between dorsolateral line and midlateral line 10–12; cephalodorsal-line scales 8; dorsal-fin rays 115–119; anal-fin rays 90–94; caudal-fin rays 10 (rarely 8 or 9); and vertebrae 9 + 47–48 = 56–57. [ABSTRACT FROM AUTHOR]

Both gonadotropin-inhibitory hormone (GnIH) and kisspeptin play a critical role in the control of reproduction in vertebrates. However, the spatial–temporal expression patterns and possible roles of these two peptides during development are poorly understood. To this end, we evaluated dynamic expression profiles of lpxrfa, the piscine ortholog of gnih, kisspeptin (kiss2) and their receptor mRNAs during embryogenesis and early larval development in a flatfish, Cynoglossus semilaevis. Quantitative RT-PCR analysis indicated that both ligand (lpxrfa and kiss2) and receptor (lpxrfa-r and kiss2r) mRNAs were detected in unfertilized eggs and during embryogenesis, but with different expression profiles. In addition, lpxrfa-r, kiss2 and kiss2r transcripts increased significantly at 6 days post-hatching (dph), while lpxrfa mRNAs did not vary significantly during all larval development. Taken together, these results of the present study suggest that both LPXRFa and kisspeptin systems could be functional and play differential roles during embryogenesis and early larval development in tongue sole. [ABSTRACT FROM AUTHOR]

The Chinese tongue sole (Cynoglossus semilaevis) is a typical female heterogamete species that exhibits female-biased sexual size dimorphism, which has severely hindered the sustainable development of the species in aquaculture. In the present study, four important somatotropic and reproductive tissues including brain, pituitary, liver, and gonad from 15 females and 15 males were used for transcriptome analysis via RNA-seq. A mean of 37,533,991 high-quality clean reads was obtained from each library and 806, 1482, 818, and 14,695 differentially expressed genes in female and male were identified from the brain, pituitary, liver, and gonad, respectively (fold change ≥ 2 and q < 0.05). Enrichment analyses of GO terms and KEGG pathways showed that nucleic acid-binding transcription factor activity, G-protein-coupled receptor activity, MAPK signaling pathway, steroid biosynthesis, and neuroactive ligand-receptor interaction may be involved in the sexual growth differences. Furthermore, via weighted gene co-expression network analyses, two modules (yellowgreen and salmon4) were identified to be significantly positive-correlated with female-biased sexual size dimorphism. An illustrated network map drawn by these two modules enabled the identification of a series of hub genes, including nipped-B-like protein A (nipbla), transcriptional activator protein Pur-beta-like (purb), and BDNF/NT-3 growth factors receptor (ntrk2). Detailed functional investigation of these networks and hub genes will further improve our understanding of the underlying molecular mechanism of sexual size dimorphism in fish. [ABSTRACT FROM AUTHOR]

Heavy metals with high bioaccumulation capacity are considered as important contaminants and may be available in high concentrations in environment and biota samples. The main aim of this study was to determine the concentration of nickel and thallium in Cynoglossus arel fish in Musa estuary. Sixty-seven fish samples were collected from Musa estuary during five intervals of 15 days in summer 2013. After biometric measurements, the concentrations of nickel and thallium were measured by graphite furnace atomic absorption spectrophotometer. The mean concentration of nickel and thallium in muscle tissue of fish samples was 2.458 ± 0.910 and 0.781 ± 1.754 mg kg/ww, respectively. The GLM analysis showed a significant negative relationship between nickel concentration and length. In addition, there was a significant positive relationship between thallium concentration and fish length. Nickel concentration exceeded the allowable standards of WHO and FDA in Cynoglossus arel. Therefore, regarding with high consumption of seafood in this region, it is recommended that these fishes should be consumed under a nutritionist counseling. [ABSTRACT FROM AUTHOR]

Measurements and observations of juvenile hatchery-reared three-lined tonguefish Cynoglossus abbreviatus were performed to examine morphological abnormalities, focusing on five asymmetric characters: eye position, body coloration, presence of a nasal tube, presence of dentition, and premaxillary length. Hatchery-reared juveniles were classified into a normal type similar to wild fish, an ambicolored type (with eyes on both sides of the body), and an intermediate type showing characteristics between the other two types. The ambicolored and intermediate types are considered metamorphosis-related malformations of flatfishes. Pseudoalbinism and reversal, which are commonly observed in hatchery-reared flatfishes, were not found in this study. The most significant result of the present study on hatchery-reared three-lined tonguefish is that the eye position and body coloration of the intermediate-type individuals were between the normal and ambicolored types. Eye migration and body coloration on the blind side of the intermediate type did not synchronize. [ABSTRACT FROM AUTHOR]

Phenotypic and genetic parameters for growth-related traits in the half-smooth tongue sole, Cynoglossus semilaevis, were estimated in 22 full-sib families produced by normal and neo-male breeding stocks. As phenotypic males with female genotypes, neo-males are harmful in C. semilaevis aquaculture because they reduce overall production. The present study evaluated the difference in the growth-related traits: total length (TL), body weight (BW) and square root of body weight (SQ_BW) at the age of 570 days between normal and neo-male offspring (neo-males used as male parents). The difference in the proportion of females between normal and neo-male offspring was also assessed. Based on the linear mixed model, restricted maximum likelihood (REML) and best linear unbiased prediction (BLUP) were used to estimate various (co)variance components and estimated breeding values (EBVs) of growth-related traits. As a result, all the mean values of the three studied traits were significantly larger in normal offspring than in neo-male offspring. Additionally, the female proportion was significantly larger in normal offspring than in neo-male offspring. Heritability was 0.128±0.066 2 for TL, 0.128±0.065 5 for BW and 0.132±0.062 9 for SQ_BW, all of which were low level heritabilities. The correlation coefficients of EBVs and phenotypic values of the target traits were 0.516 for TL, 0.524 for BW and 0.506 for SQ_BW, all of which were highly significant ( P <0.01). Genetic correlations among TL, BW and SQ_BW were positive high (0.921-0.969) and higher than those of phenotype (0.711-0.748), both of which had low standard errors (0.063-0.123 for genotype, and 0.010-0.018 for phenotype). Compared with normal offspring, neo-male offspring have lower breeding values for each studied trait through EBVs comparison. Therefore, neo-male offspring should not be used as broodstock in a C. semilaevis breeding programs. [ABSTRACT FROM AUTHOR]

Marine organisms comprising microbes, plants, invertebrates, and vertebrates elaborate an impressive array of structurally diverse antimicrobial products ranging from small cyclic compounds to macromolecules such as proteins. Some of these biomolecules originate directly from marine animals while others arise from microbes associated with the animals. It is noteworthy that some of the biomolecules referred to above are structurally unique while others belong to known classes of compounds, peptides, and proteins. Some of the antibacterial agents are more active against Gram-positive bacteria while others have higher effectiveness on Gram-negative bacteria. Some are efficacious against both Gram-positive and Gram-negative bacteria and against drug-resistant strains as well. The mechanism of antibacterial action of a large number of the chemically identified antibacterial agents, possible synergism with currently used antibiotics, and the issue of possible toxicity on mammalian cells and tissues await elucidation. The structural characteristics pivotal to antibacterial activity have been ascertained in only a few studies. Demonstration of efficacy of the antibacterial agents in animal models of bacterial infection is highly desirable. Structural characterization of the active principles present in aqueous and organic extracts of marine organisms with reportedly antibacterial activity would be desirable. [ABSTRACT FROM AUTHOR]

Follistatin (FST) is a monomeric glycoprotein highly enriched in cysteines and belongs to TGF-β superfamily. FST can suppress the secretion of follicle-stimulating hormone and plays a vital role in the reproduction of vertebrates. We used rapid amplification of cDNA ends technology to clone the FST gene of half-smooth tongue sole, Cynoglossus semilaevis. We characterized its phylogenetic context and expression patterns to elucidate its function in the breeding season. The full-length sequence of FST is 1 455 bp and encodes a protein of 321 amino acids. We investigated the expression pattern of FST in C. semilaevis at different stages of reproduction using reverse transcription-polymerase chain reaction (RTPCR). FST mRNA was expressed in all 13 tissues analyzed, and was expressed at high levels in gonad and at slightly lower levels in gill and brain. During the reproductive cycle of C. semilaevis, the transcript level of FST was the highest in the perinucleolus stage, decreased in the primary yolk stage, slightly increased in the tertiary yolk stage, and then reduced to a minimal level in the atretic follicles stage of the ovary. We concluded that FST suppressed follicle-stimulating hormone, which stimulated oocyte development. However, no significant variation was observed across all stages of testis development, although the expression level in the spermatogenesis stage was relatively low, which may result from the regulation of FST by aromatase. [ABSTRACT FROM AUTHOR]

Wnt4 gene plays a role in developmental processes in mammals. However, little is known regarding its function in teleosts. We cloned and characterized the full-length half-smooth tongue sole (Cynoglossus semilaevis) wnt4a gene (CS-wnt4a). CS-wnt4a cDNA was 1746 bp in length encoding 353aa. CS-wnt4a expression level was highest in the testis, and gradually increased in the developing gonads until 1 year of age. In situ hybridization revealed that CS-wnt4a expression level was highest in stage II oocytes and sperm in the adult ovary and testis, respectively. CS-wnt4a expression level was significantly up-regulated in the gonads after exposure to high temperature. The level of methylation of the CS-wnt4a first exon was negatively correlated with the expression of CS-wnt4a. The branch-site model suggested that vertebrate wnt4a differed significantly from that of wnt4b, and that the selective pressures differed between ancestral aquatic and terrestrial organisms. Two positively selected sites were found in the ancestral lineages of teleost fish, but none in the ancestral lineages of mammals. One positively selected site was located on the a-helices of the 3D structure, the other on the random coil. Our results are of value for further study of the function of wnt4 and the mechanism of selection. [ABSTRACT FROM AUTHOR]

Genetic sex determination by W and Z chromosomes has developed independently in different groups of organisms. To better understand the evolution of sex chromosomes and the plasticity of sex-determination mechanisms, we sequenced the whole genomes of a male (ZZ) and a female (ZW) half-smooth tongue sole (Cynoglossus semilaevis). In addition to insights into adaptation to a benthic lifestyle, we find that the sex chromosomes of these fish are derived from the same ancestral vertebrate protochromosome as the avian W and Z chromosomes. Notably, the same gene on the Z chromosome, dmrt1, which is the male-determining gene in birds, showed convergent evolution of features that are compatible with a similar function in tongue sole. Comparison of the relatively young tongue sole sex chromosomes with those of mammals and birds identified events that occurred during the early phase of sex-chromosome evolution. Pertinent to the current debate about heterogametic sex-chromosome decay, we find that massive gene loss occurred in the wake of sex-chromosome 'birth'. [ABSTRACT FROM AUTHOR]

Endocrine disrupting chemicals (EDCs) are increasingly viewed as persistent pollutants, similar to natural hormones in function. This paper describes the expression profiles of 7 genes ( DMRT, VTG GnRHR FSHR CYP17A CYP19A, and CYP19B) involved in sex steroid synthesis and action as well as sexual development in adult male and female Cynoglossus semilaevis, after exposure to different concentrations of Bisphenol A (BPA) and 17β-estradiol (E2). Both BPA (1, 10, 50, 125, and 250 mg/kg) and E2 (0.5, 5, and 10 mg/kg) induced changes in target gene expression, although the estrogenic effects of E2 as a model estrogen were stronger. Among the 7 genes, VTG CYP17A and CYP19 responded strongly to BPA or E2 exposure and can thus serve as reference biomarkers for estrogenic EDCs exposure in marine teleosts. These data will provide a window to establish a hypothalamic-pituitary-gonadal model in C. semilaevis to better understand the effect pathways of EDCs. [ABSTRACT FROM AUTHOR]

A new cell line (TSHKC) derived from half-smooth tongue sole ( Cynoglossus semilaevis) head kidney was developed. The cell line was subcultured for 40 passages over a period of 360 days. The cell line was optimally maintained in minimum essential medium supplemented with HEPES, antibiotics, fetal bovine serum, 2-Mercaptoethanol (2-Me), sodium pyruvate and basic fibroblast growth factor. The suitable growth temperature for TSHKC cells was 24 °C, and microscopically, TSHKC cells were composed of fibroblast-like cells. Chromosome analysis revealed that the TSHKC cell line had a normal diploid karyotype with 2 n = 42, contained the heterogametic W chromosome. The TSHKC cell line was found to be susceptible to lymphocystis disease virus. The fluorescent signals were observed in TSHKC when the cells were transfected with green fluorescent protein and red fluorescent protein reporter plasmids. [ABSTRACT FROM AUTHOR]

Concentrations of Cd, Co, Cu, Ni, Pb, Fe and Zn were determined in the muscle, liver and gills of three commercial benthic and pelagic fish species ( Johnius belangerii, Euryglossa orientalis and Cynoglossus arel) from three estuaries in the northwest Persian Gulf. Metals levels varied significantly depending on the tissues, species and locations. Generally, the results showed that liver accumulate higher concentrations of the metals in comparison to muscle and gills, except in few cases. Among the species, E. orientalis showed the highest levels of Co, Cu, Ni and Fe, while the highest concentrations of Pb and Zn were observed in C. arel. J. belangerii accumulated the highest level of Pb element. [ABSTRACT FROM AUTHOR]

Research was performed to investigate the concentration of mercury in muscle and liver of fish species from freshwater ( Barbus xanthopterus, Barbus grypus, Liza abu) and saltwater ( Cynoglossus arel, Periophthalmus waltoni, Otolithes ruber) in Khouzestan, Iran. In freshwater fish, muscle was polluted in comparison with liver except for Barbus xanthopterus which high levels of mercury were measured in liver. In saltwater fish liver was contaminated than muscle except for Cynoglossus arel which high level of mercury was found in muscle. Significant variations in metal values were evaluated using student's t test at P < 0.05. Mercury concentrations were well above the permissible limits suggested by WHO and FDA guidelines. [ABSTRACT FROM AUTHOR]

Major histocompatibility complex (MHC) class II B molecules play an important role in the adaptive immune response in fish. Previous study has reported that two highly polymorphic class II B genes, Cyse-DAB and Cyse-DBB exist in half-smooth tongue sole ( Cynoglossus semilaevis). In this study, the polymorphism within exon 2 of the class II B genes following bacterial challenge was evaluated. Two hundred C. semilaevis individuals were injected intraperitoneally with Vibrio anguillarum. Muscle tissue from the first 20 dead and 20 of the survivors was collected for genotyping. Sixty alleles from the 40 individuals were isolated, of which 32 belonged to Cyse-DAB and 28 belonged to Cyse-DBB. The rate of d (non-synonymous substitution) was higher than that of d (synonymous substitution) in the PBRs (peptide binding residues) of both class II B genes. Conversely, the rate of d was higher than d in the non-PBRs and the complete exon 2 sequence. Thus, the results suggest that positive selection has occurred in the PBRs and purifying selection in the non-PBRs and exon 2. Thirteen class II B alleles were used to study the association between alleles and resistance to infection. Though not significant, alleles Cyse-DAB*0601, Cyse-DAB*0706, and Cyse-DBB*0101, Cyse-DBB*1301 were only found in surviving individuals and may represent alleles that have resistance against V. anguillarum infection. Alleles Cyse-DAB*0701 and Cyse-DAB*1301 were significantly more prevalent in dead individuals than in surviving ones and may represent alleles that are associated with increased susceptibility to V. anguillarum infection. [ABSTRACT FROM AUTHOR]

Abstract  The larval distribution and feeding habits of Cynoglossus abbreviatus and Cynoglossus lighti were investigated around the Rokkaku estuary in Ariake Bay during October 2004 and March and April 2005. Cynoglossus abbreviatus started and completed metamorphosis at larger sizes than C. lighti. Developmental phases consisted primarily of individuals at metamorphosis and postmetamorphosis, which were mainly distributed inside and just outside the Rokkaku River in March–April for C. abbreviatus and October for C. lighti. Tidal changes in vertical distribution just outside the river mouth differed between the closely related species; C. abbreviatus was distributed in the surface and middle layers at flood tide, and aggregated near the bottom (where the current speed was lowest) at ebb tide. Cynoglossus lighti stayed mainly near the bottom during all tidal phases. Larvae of both Cynoglossus species selectively fed on copepods (Pseudobradya sp.), which were scarce in the waters. These facts suggest that the nursery grounds of the two Cynoglossus species were different in terms of their tidal vertical distribution patterns. [ABSTRACT FROM AUTHOR]

Twenty novel microsatellite markers for halfsmooth tongue sole (Cynoglossus semilaevis) were isolated and characterized from a simple sequence repeat-enriched library constructed with (GA)15 and (CA)15. The polymorphism was assessed using 32 individuals collected from one locality along the Chinese coast. The result showed the number of alleles ranged from 2 to 14, with an average of 8.7 alleles/locus. The values of observed and expected heterozygosities ranged from 0.2609 to 1.0000 and from 0.2367 to 0.9716, respectively. Six microsatellite loci departed significantly from Hardy-Weinberg equilibrium. No linkage disequilibrium was found. These markers will be useful for the conservation studies and population structure assessment for this species. [ABSTRACT FROM AUTHOR]

Abstract  Genetic differences between trilinear and dilinear forms of a tongue sole, Cynoglossus interruptus, were examined by use of isozymes. Unequivocal differences were detected between the two forms, including complete replacement of alleles at the FBALD-2*, MDH-3*, PROT-1*, and SOD-1* loci, almost complete replacement at the AAT-3*, AH-1*, AH-2*, EST-3*, G3PDH-3*, GPI-2*, IDHP-1*, and MDH-1* loci, and extreme differences in allelic frequencies at the GPI-1* and PGDH* loci. The genetic distances (D values) between the two forms were 0.4207–0.4353, figures predicatively significant at the specific level. The considerable genetic differences strongly suggested that the two forms represent distinct species. [ABSTRACT FROM AUTHOR]