56 research outputs found

    Generation of Biotechnology-Derived Flavobacterium columnare Ghosts by PhiX174 Gene E-Mediated Inactivation and the Potential as Vaccine Candidates against Infection in Grass Carp

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    Flavobacterium columnare is a bacterial pathogen causing high mortality rates for many freshwater fish species. Fish vaccination with a safe and effective vaccine is a potential approach for prevention and control of fish disease. Here, in order to produce bacterial ghost vaccine, a specific Flavobacterium lysis plasmid pBV-E-cat was constructed by cloning PhiX174 lysis gene E and the cat gene with the promoter of F. columnare into the prokaryotic expression vector pBV220. The plasmid was successfully electroporated into the strain F. columnare G4cpN22 after curing of its endogenous plasmid. F. columnare G4cpN22 ghosts (FCGs) were generated for the first time by gene E-mediated lysis, and the vaccine potential of FCG was investigated in grass carp (Ctenopharyngodon idellus) by intraperitoneal route. Fish immunized with FCG showed significantly higher serum agglutination titers and bactericidal activity than fish immunized with FKC or PBS. Most importantly, after challenge with the parent strain G4, the relative percent survival (RPS) of fish in FCG group (70.9%) was significantly higher than FKC group (41.9%). These results showed that FCG could confer immune protection against F. columnare infection. As a nonliving whole cell envelope preparation, FCG may provide an ideal alternative to pathogen-based vaccines against columnaris in aquaculture

    Multidose Streptozotocin Induction of Diabetes in BALB/c Mice Induces a Dominant Oxidative Macrophage and a Conversion of T(H)1 to T(H)2 Phenotypes During Disease Progression

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    Macrophages (Mp) are implicated in both early and late phases in type 1 diabetes development. Recent study has suggested that a balance between reductive Mp (RMp) and oxidative Mp (OMp) is possible to regulate T(H)1/T(H)2 balance. The aim of this study is to investigate the redox status of peritoneal Mp and its cytokine profile during the development of autoimmune diabetes induced by multiple low-dose streptozotocin in BALB/c mice. Meanwhile, the polarization of T(H)1/T(H)2 of splenocytes or thymocytes was also examined. We found that peritoneal Mp appeared as an “incomplete” OMp phenotype with decreased icGSH along with disease progression. The OMp showed reduced TNF-α, IL-12, and NO production as well as defective phagocytosis activity compared to nondiabetic controls; however, there was no significant difference with IL-6 production. On the other hand, the levels of IFN-γ or IL-4 of splenocytes in diabetic mice were significantly higher compared to the control mice. The ratio of IFN-γ to IL-4 was also higher at the early stage of diabetes and then declined several weeks later after the occurrence of diabetes, suggesting a pathogenetic T(H)1 phenotype from the beginning gradually to a tendency of T(H)2 during the development of diabetes. Our results implied that likely OMp may be relevant in the development of type 1 diabetes; however, it is not likely the only factor regulating the T(H)1(H)/T(H)2 balance in MLD-STZ-induced diabetic mice

    The association of two single nucleotide polymorphisms (SNPs) in growth hormone (GH) gene with litter size and superovulation response in goat-breeds

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    Two active mutations (A 781 G and A 1575 G) in growth hormone (GH) gene, and their associations with litter size (LS), were investigated in both a high prolificacy (Matou, n = 182) and a low prolificacy breed (Boer, n = 352) by using the PCR-RFLP method. Superovulation experiments were designed in 57 dams, in order to evaluate the effect of different genotypes of the GH gene on superovulation response. Two genotypes (AA and AB, CC and CD) in each mutation were detected in these two goat breeds. Neither BB nor DD homozygous genotypes were observed. The genotypic frequencies of AB and CC were significantly higher than those of AA and CD. In the third parity, Matou dams with AB or CC genotypes had significantly larger litter sizes than those with AA and CD (p < 0.05). On combining the two loci, both Matou and Boer dams with ABCD genotype had the largest litter sizes when compared to the other genotypes (p < 0.05). When undergoing like superovulation treatments, a significantly higher number of corpora lutea and ova, with a lower incidence of ovarian cysts, were harvested in the AB and CC genotypes than in AA and CD. These results show that the two loci of GH gene are highly associated with abundant prolificacy and superovulation response in goat breeds

    Dual inhibition of AKT‐mTOR and AR signaling by targeting HDAC3 in PTEN‐ or SPOP‐mutated prostate cancer

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    Abstract AKT‐mTOR and androgen receptor (AR) signaling pathways are aberrantly activated in prostate cancer due to frequent PTEN deletions or SPOP mutations. A clinical barrier is that targeting one of them often activates the other. Here, we demonstrate that HDAC3 augments AKT phosphorylation in prostate cancer cells and its overexpression correlates with AKT phosphorylation in patient samples. HDAC3 facilitates lysine‐63‐chain polyubiquitination and phosphorylation of AKT, and this effect is mediated by AKT deacetylation at lysine 14 and 20 residues and HDAC3 interaction with the scaffold protein APPL1. Conditional homozygous deletion of Hdac3 suppresses prostate tumorigenesis and progression by concomitant blockade of AKT and AR signaling in the Pten knockout mouse model. Pharmacological inhibition of HDAC3 using a selective HDAC3 inhibitor RGFP966 inhibits growth of both PTEN‐deficient and SPOP‐mutated prostate cancer cells in culture, patient‐derived organoids and xenografts in mice. Our study identifies HDAC3 as a common upstream activator of AKT and AR signaling and reveals that dual inhibition of AKT and AR pathways is achievable by single‐agent targeting of HDAC3 in prostate cancer

    Two new species of Xyalidae (Monhysterida, Nematoda) from the East China Sea

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    Wang, Chunming, An, Liguo, Huang, Yong (2018): Two new species of Xyalidae (Monhysterida, Nematoda) from the East China Sea. Zootaxa 4514 (4): 583-592, DOI: https://doi.org/10.11646/zootaxa.4514.4.1

    Daptonema donghaiensis Wang & An & Huang 2018, sp. nov.

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    Daptonema donghaiensis sp. nov. (Figs 1-2) Type material. Five males and two females were measured and studied. Holotype male and paratype male on slide DH12-422-13; paratype females on slides DH12-422-8, DH12-422-11; additional males on slides DH12-413-2, DH12-418-4, DH12-418-5. Type locality and habitat. Sublittoral in the East China Sea. Holotype and paratype specimens comes from Station DH4-2: 29°18´N, 123° 22´E, water depth 70 m. Muddy with a little sand sediment fraction. Additional specimens come from station DH4-1: 29° 28´N, 123° 6´E, water depth 61 m. Muddy with a little sand sediment fraction. Etymology. This name refers to the East China Sea, where the species was collected. ‘Donghai’ means ‘East China Sea’ in Chinese. Measurements. Table 1. Description. Male. Body short, 836–972 µm long. Maximum diameter 27–34 µm. Cuticle finely striated. Epidermal chords consisting of transparent cells present in dorsal, ventral and lateral sides of anterior and middle body parts, especially in pharyngeal region. Buccal cavity funnel-shaped, 5.2–5.7 µm wide and 4.1–4.5 µm deep. Six inner labial papilliform sensilla. Six outer labial setae (9 µm) and four cephalic setae (6.4–7.2 µm) arranged in one circle. Somatic setae absent. Amphids circular, 6.6–6.9 µm in diameter (43.5–47.1% of corresponding body diameter) and 13–15.8 µm (1.7–2.2 times the head diameter) from the anterior body end. Pharynx muscular and cylindrical, 144–172 µm long, 16–20% of body length. Nerve ring 66–94 µm from the anterior body end. Cardia conical, 5.8–6 µm long. Tail conico-cylindrical, 113–136 µm long (5–6.5 times the anal body diameter), cylindrical part long (46-54% of tail length). Three caudal glands in line. Two terminal setae, 6.5–7.5 µm long. Gonads diorchic and opposed. Anterior testis to the left of the intestine. Posterior testis to the right of the intestine. Spicules L-shaped, 26–31 µm along arc (1.33–1.48 times the anal body diameter), cephalated at proximal end. Gubernaculums tubular, parallel to the distal part of spicules, 9.5 µm long. Female. Similar to males in most respects. Reproductive system monodelphic, prodelphic. Ovary outstretched, situated to the right of the intestine. Spermatheca oval in shape (43 µm long, 22 µm wide). Vagina short, bending anteriorly. Vulva a transverse slit-like structure, located slightly posterior to midbody, 544 µm (63.6%) from anterior end. Differentiation diagnosis and discussion. Daptonema donghaiensis sp. nov. is characterized by a short body, finely striated cuticle, six outer labial setae and four cephalic setae in one circle, absence of somatic setae, epidermal chords of transparent cells in anterior and middle body parts, spicules L-shaped and cephalated at proximal end, gubernaculums tubular, tail conico-cylindrical with a long cylindrical part. Daptonema possesses the highest number of valid species in Xyalidae and is taxonomically one of the most difficult genera. The presence of transparent cells in epidermal chords is considered one of the diagnostic features by Aryuthaka & Kito (2012). To date, four species of Daptonema possessing transparent cells have been reported: D. trabeculosum G. Schneider, 1906; D. conicum Filipjev, 1922; D. hyalocella Aryuthaka & Kito, 2012 and D. setihyalocella Aryuthaka & Kito, 2012 (Aryuthaka & Kito 2012). D. donghaiensis sp. nov. mostly resembles D. hyalocella in its similar body length, finely striated cuticle, tail with a long cylindrical part and two terminal setae. However, it differs in its slender body (maximum diameter 27– 35 µm vs. 36–48 µm), anterior sensilla configuration (6+10 vs. 6+12), narrower head diameter (6.5–9.5 µm vs. 13.5–15.3 µm), larger amphids (35–47% of corresponding body diameter vs. 25–33% of corresponding body diameter), position of amphids (1.7–2.2 times the head diameter vs. 0.5–0.9 times the head diameter, from anterior end), shape of spicules (L-shaped with cephalated proximal end vs. loosely S-shaped without proximal cephalation) and distinct tubular gubernaculums (indistinct in D. hyalocella) (Aryuthaka & Kito 2012). D. donghaiensis sp. nov. differs from D. setihyalocella in its body length (786–972 µm vs. 1124–1347 µm), absence of somatic setae, anterior sensilla configuration (6+10 vs. 6+12), position of amphids (1.7–2.2 times the head diameter vs. 0.6–0.9 times the head diameter, from anterior end), tail length (5–6.5 times the anal body diameter vs. 3.6–4.1 times the anal body diameter), length of spicules (26–31 µm vs. 48–57 µm), and gubernaculums without apophysis (Aryuthaka & Kito 2012). D. donghaiensis sp. nov. differs from D. trabeculosum in its shorter body (786–972 µm vs. 880–1100 µm), anterior sensilla configuration (6+10 vs. 6+12), position of amphids (1.7–2.2 times the head diameter vs. 0.9 times the head diameter), shorter spicules (26–31 µm vs. 34–36 µm), gubernaculum shape (tubular vs. gubernaculum with lateral piece) and absence of somatic setae. D. donghaiensis sp. nov. resembles D. conicum in its similar body length, cephalic setae configuration and similar values of de Man’s c ratios. However, it differs in its slender body (maximum diameter 27–35 µm vs. 36–43 µm), longer cephalic setae (6.4 µm vs. 2 µm), larger amphids (6–7.7 µm vs. 4 µm), position of amphids (1.7–2.2 times head diameter vs. 1.4 times the head diameter, from anterior end) and shorter gubernaculums (9.5 µm vs. 17 µm) (Lorenzen 1977).Published as part of Wang, Chunming, An, Liguo & Huang, Yong, 2018, Two new species of Xyalidae (Monhysterida, Nematoda) from the East China Sea, pp. 583-592 in Zootaxa 4514 (4) on pages 584-587, DOI: 10.11646/zootaxa.4514.4.11, http://zenodo.org/record/260929

    Cobbia heterospicula Wang & An & Huang 2018, sp. nov.

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    Cobbia heterospicula sp. nov. (Figs 3-4) Type material. Four males and one female were measured and studied. Holotype male on slide DH12-387-5; female on slide DH12-332-1; additional males on slides DH12-434-2, DH12-433-2, DH12-513-2. Type locality and habitat. Sublittoral zone in the East China Sea. Holotype male comes from station DH3-8, 30° N, 126° E, water depth of 79 m. Muddy with a small fraction of sand sediment. Additional specimens come from station DH3-3, 30°N, 123°29´E, water depth 69 m; DH4-3, 29° 5´N, 123°47´E, water depth 79 m; DH5-1, 28° 27´N, 122°10.4´E, water depth 43 m. Muddy with a little sand sediment fraction. Etymology. The species name refers to the unequal size of the spicules. Measurements. Table 2. Description. Male. Body short and slender, 987–1094 µm long. Maximum diameter 16–19 µm. Anterior body slightly truncated and posterior body filiform. Buccal cavity funnel-shaped, with one slightly cuticularized dorsal tooth and two small subventral teeth. Head blunt, weakly set off. Six inner labial setae setiform. Six outer labial setae (7–9.4 µm) and four cephalic setae (4–4.5 µm) arranged in one circle. Somatic setae absent. Amphids circular, 4.7–6.5 µm in diameter (41–60% of corresponding body diameter), 21–29 µm (4.5–6.5 times the head diameter) from the anterior body end. Pharynx cylindrical, 142–152 µm long, posterior part widens but without forming a real bulb. Nerve ring 58–83 µm from the anterior body end, 40–54% of the pharynx length. Excretory glands and excretory pore unobserved. Cardia conical, 5 µm long. Tail conico-cylindrical, 227–307 µm long (13– 18 times the anal body diameter), cylindrical part filiform (167–245 µm, 74–80% of tail length). Gonads diorchic and opposed. Anterior testis to the left of the intestine and posterior testis to the right of the intestine. Spicules curved, paired but differing in length, right spicule 27–33 µm and left spicule 15–20 µm (measured along arc). Gubernaculums are parallel to the distal part of spicules, with tapering dorsal apophysis. Female. Similar to males in most respects, but with a longer tail. Reproductive system monodelphic, prodelphic. A single ovary, anteriorly outstretched, situated to the right of the intestine. Vagina short, bending anteriorly. Spermatheca not seen. Vulva a transverse slit-like structure, located at approximately the midbody, 603 µm (50.4%) from the anterior body end. Differentiation diagnosis and discussion. Cobbia heterospicula sp. nov. is characterized by a short and slender body, buccal cavity with one dorsal and two small subventral teeth, amphids located far from the anterior body end, conico-cylindrical tail with a long filiform portion, paired spicules of different sizes, and gubernaculums with dorsal apophysis. C. heterospicula sp. nov. can be easily distinguished from the other species by its unequally sized spicules and slender body. Anterior sensilla configuration and tooth composition are two diagnostic features in Cobbia (Lo Russo & Pastor de Ward 2012), and this new species belongs to the group of species that have anterior sensilla configuration of 6+10, one large dorsal tooth and two subventral teeth. C. heterospicula sp. nov. resembles C. caledonia in its body length, anterior sensilla configuration, large amphids (0.41–0.6 times the corresponding body diameter), and position of amphids (more than two times the head diameter from the anterior body end) but differs in its spicules without proximal cephalation, gubernaculums with dorsal apophysis and tail shape (conico-cylindrical with a long filiform portion vs. conical) (Warwick & Platt 1973). Gubernaculum apophysis was observed in C. heterospicula sp. nov., C. truncata and C. sinica. However, the apophysis in C. sinica is small, and in C. truncata, the distal end is blunt or slender (Huang & Zhang 2010, Wieser 1959). The new species differs from C. truncata in its shorter body length (1.06–1.19 mm vs. 1.45–1.59 mm), cuticle pattern (finely striated vs. coarsely annulated), anterior sensilla configuration (6+10 vs. 6+12), teeth composition (one dorsal and two small subventral teeth vs. three teeth of equal size), absence of cervical setae and somatic setae, and longer tail (17 anal body diameters long vs. 5.8 anal body diameters long) (Wieser 1959).Published as part of Wang, Chunming, An, Liguo & Huang, Yong, 2018, Two new species of Xyalidae (Monhysterida, Nematoda) from the East China Sea, pp. 583-592 in Zootaxa 4514 (4) on pages 587-591, DOI: 10.11646/zootaxa.4514.4.11, http://zenodo.org/record/260929

    Inactivation of mitogen-activated protein kinase signaling pathway reduces caspase-14 expression in impaired keratinocytes

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    Objective(s):Several investigations have revealed that caspase-14 is responsible for the epidermal differentiation and cornification, as well as the regulation of moisturizing effect. However, the precise regulation mechanism is still not clear. This study was aimed to investigate the expression of caspase-14 in filaggrin-deficient normal human epidermal keratinocytes (NHEKs) and to explore the possible mechanism that contributes to the regulation of caspase-14. Materials and Methods:The filaggrin-deficient NHEKs were induced by transfection with lentivirus (LV) vector encoding small hairpin RNAs (shRNA). The inhibitors SB203580, PD98059 and SP600125 were used for suppressing the expression of p38 mitogen-activated protein kinase (MAPK), p44/42 MAPK and stress-activated protein kinase/c-Jun N-terminal kinase (SAPK/JNK). The expression of filaggrin, p38 MAPK, p44/42 MAPK and SAPK/JNK, caspase-14, keratin1and keratin2 were detected by western blot.  Results:In filaggrin-deficient NHEKs, the expression of p38, p44/42 MAPK and SAPK/JNK and caspase-14 were significantly decreased. The inhibition of p38 and SAPK/JNK reduced the expression of caspase-14, while the p44/42 MAPK showed no consistent effects. Moreover, the filaggrin knockdown decreased the expression of keratin2, but had no effects on the level of keratin1. Conclusion: The decreased expression of caspase-14 in filaggrin-deficient NHEKs may be induced by the inactivation of MAPK signaling pathway. These provide a novel perspective to understand the mechanism for the protective effects of filaggrin and caspase-14 on skin barrier function
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