The insulin-loaded alginate-chitosan microspheres prepared by SPG membrane emulsification and two-step solidification could realize the pH-sensitive release of insulin.
The alginate–chitosan ...microspheres with narrow size distribution were prepared by membrane emulsification technique in combination with ion (Ca
2+) and polymer (chitosan) solidification. The preparation procedure was observed, and the physical properties (particle size distribution, surface morphology, chitosan distribution, zeta potential) of the microspheres were characterized. Subsequently, the microspheres were employed to load model peptide of insulin. The effect of loading ways on the loading efficiency and immunological activity of insulin were investigated. It was shown that the higher loading efficiency (56.7%) and remarkable activity maintenance (99.4%) were obtained when the insulin was loaded during the chitosan solidification process (Method B). Afterward, the release profile
in vitro for the optimal insulin-loaded microspheres was investigated. Under the pH conditions of gastrointestinal environment, only 32% of insulin released during the simulated transit time of drug (2
h in the stomach and 4
h in the intestinal). While under the pH condition of blood environment, insulin release was stable and sustained for a long time (14
days). Furthermore, the chemical stability of insulin released from the microspheres was well preserved after they were treated with the simulated gastric fluid containing pepsin for 2
h. Finally, the blood glucose level of diabetic rats could be effectively reduced and stably kept for a long time (∼60
h) after oral administration of the insulin-loaded alginate–chitosan microspheres. Therefore, the alginate–chitosan microspheres were found to be promising vectors showing a good efficiency in oral administration of protein or peptide drugs.
MicroRNAs are regulatory molecules that can be packaged into exosomes to modulate cellular response of recipients. While the role of exosomes during viral infection is beginning to be appreciated, ...the involvement of exosomal miRNAs in immunoregulation in invertebrates has not been addressed. Here, we observed that exosomes released from WSSV-injected mud crabs could suppress viral replication by inducing apoptosis of hemocytes. Besides, miR-137 and miR-7847 were found to be less packaged in mud crab exosomes during viral infection, with both miR-137 and miR-7847 shown to negatively regulate apoptosis by targeting the apoptosis-inducing factor (AIF). Our data also revealed that AIF translocated to the nucleus to induce DNA fragmentation, and could competitively bind to HSP70 to disintegrate the HSP70-Bax (Bcl-2-associated X protein) complex, thereby activating the mitochondria apoptosis pathway by freeing Bax. The present finding therefore provides a novel mechanism that underlies the crosstalk between exosomal miRNAs and apoptosis pathway in innate immune response in invertebrates.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Three Cu-Al alloys with nominal Al contents of 2.2, 4.5 and 6.9 wt% were subjected to surface mechanical attrition treatment (SMAT) to investigate the effect of stacking fault energy (SFE) on the ...microstructural feature of gradient structure (GS) layers produced by SMAT and back stress hardening during Bauschinger unloading-reloading tests. In the GS layer, the alloys with different SFE exhibit various grain morphologies, grain size distributions and densities and distributions of geometrically necessary dislocations (GNDs). Because of these differences in the microstructural features of the GS layers, the alloys show different variations associated with SMAT during the Bauschinger unloading-reloading tests, i.e. the increment of true stress decreases sharply with a decrease of SFE at small strains, while it slightly decreases at large strains. It is found that SFE can significantly affect grain boundary strengthening and back stress hardening to synthetically improve the tensile strength of GS Cu-Al alloys to different levels.
Elongases of very long-chain fatty acids (Elovls) catalyze the rate-limiting step of the elongation pathway that results in net two‑carbon elongation of pre-existing fatty acyl chains. As a set of ...crucial enzymes involved in the long-chain polyunsaturated fatty acids (LC-PUFA) biosynthesis, Elovls of fish have been investigated extensively in recent years. In the present study, we first identified two novel fish-specific elovl genes (named as elovl8a and elovl8b) from the herbivorous marine teleost rabbitfish (Siganus canaliculatus) by genomic survey and molecular cloning methods. Subsequently, their functional characteristics, tissue distribution patterns and transcriptional changes in response to different nutritional states were investigated. Full-length coding sequences of the elovl8a and elovl8b genes were 804 and 792 bp, encoding 267 and 263 amino acids, respectively. Multiple alignment, genomic synteny and phylogenetic analyses further suggested that elovl8 genes were unique to teleosts. Functional characterization by heterologous expression in yeast showed that Elovl8b could elongate C18 (18:2n-6, 18:3n-3 and 18:4n-3) and C20 (20:4n-6 and 20:5n-3) polyunsaturated fatty acids (PUFAs) to longer-chain polyunsaturated fatty acids (LC-PUFAs) whereas Elovl8a lacked this ability. In vitro, the expression of elovl8b but not elovl8a in rabbitfish hepatocytes was significantly up-regulated by incubation with 18:2n-6, 18:3n-3, 20:4n-6 and 20:5n-3, respectively. In vivo, compared with fish oil, dietary vegetable oil enriched in C18 PUFA enhanced the expression of elovl8b in rabbitfish brain, liver, intestine and gill tissues. These findings suggest that elovl8b but not elovl8a is a novel active member of the Elovl protein family involved in the LC-PUFAs biosynthesis pathway in rabbitfish, and provide novel insight into the mechanisms of LC-PUFAs biosynthesis in teleost.
•Two novel fish-specific elovl8 genes were first identified in Siganus canaliculatus.•Comprehensive studies revealed two elovl8 isoforms were widely existed in teleost.•Rabbitfish elovl8a was highly expressed in heart and spleen, while elovl8b was mainly distributed in the brain and eye.•Elovl8b but not Elovl8a was found to have the ability to elongate the LC-PUFA.•A new mechanism involved in LC-PUFA biosynthesis was established.
Display omitted
•SOM was mostly derived from plant litter rather than microbial residues.•SOM was associated with vermiculite via surface adsorption.•SOM was associated with illite and kaolinite ...through pore entrapment.•More plant carbohydrates were associated with 2:1 rather than 1:1 clay minerals.•SOM formation efficiency was affected by clay mineral type more than by plant litter type.
Plant litters incorporated in soils are decomposed by microorganisms and partially transformed into soil organic matter (SOM) through mineral-organic association and physical protection in soil aggregates. Few studies have linked the effects of clay mineralogy and plant litter quality on controlling the formation efficiency of SOM. Using model soils, the objectives of this study were (1) to determine the effects of clay mineral type and plant litter quality on soil respiration dynamics, and formation efficiency of SOM, physical fractions, and chemical and microbial compositions of SOM at the end of a 120-day incubation; (2) to unravel SOM protection mechanisms and extents by specific clay minerals; and (3) to understand the key role of clay minerals relative to plant litter quality in controlling SOM formation. The changes in X-ray diffraction peak intensity (in terms of peak height) of the clay minerals during incubation and after H2O2 treatment provided evidence for surface adsorption by vermiculite and pore entrapment by kaolinite and illite assemblages. The SOM protection extent parameter, defined based on accumulative soil respiration dynamics, explained well the variation of the formation efficiency of mineral associated SOM (MAOM) and, to a lesser extent, that of occluded particulate SOM (oPOM) in aggregates. 90–96% of plant litter-derived C was protected in the vermiculite material and 33–60% in the pure kaolinite and illite materials. The pure vermiculite material showed the greatest fractions of MAOM and oPOM, the highest relative abundances of O–alkyls and anomeric from carbohydrates in the MAOM fraction. < 1% of plant litter-derived C was transformed into microbial biomass and residues, and fungal residues only were associated with the pure vermiculite material. These results suggested that plant litter incorporated into the soil was decomposed mainly through the ex vivo modification pathway and transformed into SOM through both mineral-organic association and physical protection pathways. Plant litter-derived C was likely protected through mineral-organic association from the earlier stages of decomposition by vermiculite than kaolinite and illite, resulting in more plant carbohydrates associated with vermiculite. Therefore, clay minerals determined SOM formation pathways and played a greater role in controlling SOM formation efficiency than plant litter quality.
Mud crab, Scylla paramamosain, a euryhaline crustacean species, mainly inhabits the Indo-Western Pacific region. Wild mud crab spawn in high-salt condition and the salinity reduced with the growth of ...the hatching larvae. When the larvae grow up to megalopa, they migrate back to estuaries and coasts in virtue of the flood tide, settle and recruit adult habitats and metamorphose into the crablet stage. Adult crab can even survive in a wide salinity of 0-35 ppt. To investigate the mRNA profile after salinity stress, S. paramamosain megalopa were exposed to different salinity seawater (low, 14 ppt; control, 25 ppt; high, 39 ppt).
Firstly, from the expression profiles of Na+/K+/2Cl- cotransporter, chloride channel protein 2, and ABC transporter, it turned out that the 24 h might be the most influenced duration in the short-term stress. We collected megalopa under different salinity for 24 h and then submitted to mRNA profiling. Totally, 57.87 Gb Clean Data were obtained. The comparative genomic analysis detected 342 differentially expressed genes (DEGs). The most significantly DEGs include gamma-butyrobetaine dioxygenase-like, facilitated trehalose transporter Tret1, sodium/potassium-transporting ATPase subunit alpha, rhodanese 1-like protein, etc. And the significantly enriched pathways were lysine degradation, choline metabolism in cancer, phospholipase D signaling pathway, Fc gamma R-mediated phagocytosis, and sphingolipid signaling pathway. The results indicate that in the short-term salinity stress, the megalopa might regulate some mechanism such as metabolism, immunity responses, osmoregulation to adapt to the alteration of the environment.
This study represents the first genome-wide transcriptome analysis of S. paramamosain megalopa for studying its stress adaption mechanisms under different salinity. The results reveal numbers of genes modified by salinity stress and some important pathways, which will provide valuable resources for discovering the molecular basis of salinity stress adaptation of S. paramamosain larvae and further boost the understanding of the potential molecular mechanisms of salinity stress adaptation for crustacean species.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
PIWI-interacting RNAs (piRNAs) are abundantly found in germ cells and involved in gametogenesis and gonadal development. Information on the regulatory roles of piRNAs in crustacean reproduction, ...however, is scarce. Thus, we identified gonadal piRNAs of mud crab Scylla paramamosain. Of the 115,491 novel piRNAs, 596 were differentially expressed. Subsequently, 389,887 potential piRNA-target genes were predicted. The expression of 4 piRNAs and 9 genes with high piRNA interactions were validated with the inclusion of additional immature specimens, including LRP2 that is involved in growth and reproduction, MDN1 in ribosome biogenesis pathway and gametogenesis, and PRKDC, a DNA repair gene involved in gonadal differentiation and maturation. KEGG analysis further revealed the involvement of predicted piRNA target genes in gametogenesis- and reproduction-related pathways. Our findings provide baseline information of mud crab piRNAs and their differential expression between testes and ovaries suggests that piRNAs play an essential role in regulating gametogenesis and gonadal development.
•596 out of 115,491 novel piRNAs were gonadal differentially expressed.•Predicted potential target genes of piRNAs were involved in gametogenesis- and reproduction-related pathways.•piRNAs potentially regulate gametogenesis and gonadal development in crustaceans.
•A novel AMP TGH2 was screened from C-terminal sequence of T. granosa hemoglobin.•The MIC of TGH2 on E. coli decreased by 4-fold under ultrasound treatment.•The permeability on E. coli cell membrane ...changed with ultrasound treatment.•TGH2 had a slight untwisting effect under ultrasound treatment for 0.5 h.
In recent years, foodborne diseases caused by Escherichia coli are a major threat to the food industry and consumers. Antimicrobial peptides (AMPs) and ultrasound both have good inhibitory effects on E. coli. In this work, the mechanism of action and synergistic effect of an in silico predicted AMP, designated as TGH2 (AEFLREKLGDKCTDRHV), from the C-terminal sequence of Tegillarca granosa hemoglobin, combined with low-intensity ultrasound was explored. The minimal inhibitory concentration (MIC) of TGH2 on E. coli decreased by 4-fold to 31.25 μg/mL under 0.3 W/cm2 ultrasound treatment, while the time kill curve analysis showed that low-intensity ultrasound combined with peptide TGH2 had an enhanced synergistic bactericidal effect after 0.5 h. The permeability on E. coli cell membrane increased progressively during combined treatment with peptide TGH2 and low-intensity ultrasound, resulting in the leakage of intracellular solutes, as shown by transmission electron microscopy (TEM). Structural analysis using circular dichroism (CD) revealed that peptide TGH2 has an α-helical structure, showing a slight untwisting effect under 0.3 W/cm2 ultrasound treatment for 0.5 h. The findings here provide new insight into the potential application of ultrasound and AMPs combination in food preservation.
Mud crabs, Scylla spp., are commercially important large-size marine crustaceans in the Indo-West Pacific region. As females have the higher growth rate and economic value, the production of all ...female stocks is extremely essential in aquaculture. However, the sex determination mechanism is still unclear. Development of sex-specific genetic markers based on next-generation sequencing proved to be an effective tool for discovering sex determination system in various animals.
Restriction-site associated DNA sequencing (RAD-seq) was employed to isolate sex-specific SNP markers for S. paramamosain. A total of 335.6 million raw reads were obtained from 20 individuals, of which 204.7 million were from 10 females and 130.9 million from 10 males. After sequence assembly and female-male comparison, 20 SNP markers were identified to be sex-specific. Furthermore, ten SNPs in a short sequence (285 bp) were confirmed heterozygous in females and homozygous in males in a large population by PCR amplification and sequencing. Subsequently, a female-specific primer was successfully designed according to the female-specific nucleotide which could amplify an expected band from females but not from males. Thus, a rapid and effective method for molecular sexing in S. paramamosain was developed, meanwhile, this method could successfully identify the sex of S. tranquebarica and S. serrata. Finally, nine and four female-specific SNP markers were detected in S. tranquebarica and S. serrata, respectively.
Sex-specific SNP markers were firstly identified in crab species and showed female heterogamety and male homogamety, which provided strong genetic evidence for a WZ/ZZ sex determination system in mud crabs S. paramamosain, S. tranquebarica and S. serrata. These findings will lay a solid foundation for the study of sex determination mechanism, sex chromosome evolution, and the development of mono-sex population in crustaceans.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Innate immunity is the only defense system for resistance against infections in crustaceans. In crustaceans, white spot diseases caused by white spot syndrome virus (WSSV) are a serious viral disease ...with high accumulative mortality after infection. Attachment and entry into cells have been known to be two initial and important steps in viral infection. However, systematic information about the mechanisms related to WSSV infection in crustaceans is still limited. Previous studies have reported that cellular receptors are important in the innate immune system and are responsible for the recognition of foreign microorganisms and in the stimulation of the immune responses during infections. In this review, we summarize the current understanding of the functions of cellular receptors, including Toll, C-type lectin, scavenger receptor, β-integrin, polymeric immunoglobulin receptor, laminin receptor, globular C1q receptor, lipopolysaccharide-and β-1,3-glucan-binding protein, chitin-binding protein, Ras-associated binding, and Down syndrome cell adhesion molecule in the innate immune defense of crustaceans, especially shrimp and crabs, in response to WSSV infection. The results of this study provide information on the interaction between viruses and hosts during infections, which is important in the development of preventative strategies and antiviral targets in cultured aquatic animals.
PDF
