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International Journal of Molecular... May 2024NPC intracellular cholesterol transporter 1 (NPC1) plays an important role in sterol metabolism and transport processes and has been studied in many vertebrates and some...
NPC intracellular cholesterol transporter 1 (NPC1) plays an important role in sterol metabolism and transport processes and has been studied in many vertebrates and some insects, but rarely in crustaceans. In this study, we characterized NPC1 from () and evaluated its functions. Its total cDNA length was 4283 bp, encoding for 1344 amino acids. It contained three conserved domains typical of the NPC family (NPC1_N, SSD, and PTC). In contrast to its role in insects, was mainly expressed in the adult female hepatopancreas, with moderate expression in the ovary and heart. No expression was found in the embryo (stages CS-ZS) and only weak expression in the larval stages from hatching to the post-larval stage (L1-PL15). expression was positively correlated with ovarian maturation. In situ hybridization showed that it was mainly located in the cytoplasmic membrane and nucleus of oocytes. A 25-day RNA interference experiment was employed to illustrate the Mn-NPC1 function in ovary maturation. Experimental knockdown of using dsRNA resulted in a marked reduction in the gonadosomatic index and ecdysone content of females. The experimental group showed a significant delay in ovarian maturation and a reduction in the frequency of molting. These results expand our understanding of NPC1 in crustaceans and of the regulatory mechanism of ovarian maturation in .
Topics: Animals; Female; Palaemonidae; Ovary; Molting; Phylogeny; Amino Acid Sequence; Gene Expression Regulation, Developmental; Arthropod Proteins; RNA Interference
PubMed: 38892237
DOI: 10.3390/ijms25116049 -
International Journal of Molecular... May 2024Carotenoid cleavage oxygenases can cleave carotenoids into a range of biologically important products. Carotenoid isomerooxygenase (NinaB) and β, β-carotene 15,...
Carotenoid cleavage oxygenases can cleave carotenoids into a range of biologically important products. Carotenoid isomerooxygenase (NinaB) and β, β-carotene 15, 15'-monooxygenase (BCO1) are two important oxygenases. In order to understand the roles that both oxygenases exert in crustaceans, we first investigated () and () within the genome of Chinese mitten crab (). Their functions were then deciphered through an analysis of their expression patterns, an in vitro β-carotene degradation assay, and RNA interference. The results showed that both and contain an RPE65 domain and exhibit high levels of expression in the hepatopancreas. During the molting stage, exhibited significant upregulation in stage C, whereas showed significantly higher expression levels at stage AB. Moreover, dietary supplementation with β-carotene resulted in a notable increase in the expression of and in the hepatopancreas. Further functional assays showed that the expressed in underwent significant changes in its color, from orange to light; in addition, its β-carotene cleavage was higher than that of . After the knockdown of or in juvenile , the expression levels of both genes were significantly decreased in the hepatopancreas, accompanied by a notable increase in the redness () values. Furthermore, a significant increase in the β-carotene content was observed in the hepatopancreas when mRNA was suppressed, which suggests that plays an important role in carotenoid cleavage, specifically β-carotene. In conclusion, our findings suggest that and may exhibit functional co-expression and play a crucial role in carotenoid cleavage in crabs.
Topics: Animals; beta Carotene; Brachyura; beta-Carotene 15,15'-Monooxygenase; Hepatopancreas; Molting; Oxygenases; Phylogeny; Arthropod Proteins
PubMed: 38891781
DOI: 10.3390/ijms25115592 -
Animals : An Open Access Journal From... May 2024Fasting-induced molting (FIM) is a common method used to improve the laying performance of aged laying hens. Nevertheless, this approach may impose various stresses on...
Fasting-induced molting (FIM) is a common method used to improve the laying performance of aged laying hens. Nevertheless, this approach may impose various stresses on chickens, such as disruptions in intestinal flora and inflammation issues within the intestines. However, the impact of an imbalance in intestinal flora on intestinal health during the FIM process remains elusive. Therefore, intestinal injury, the microbiome, and the metabolome were analyzed individually and integrated to elucidate the impact of the intestinal flora on intestinal health during the FIM process. The findings indicated that fasting resulted in a notable reduction in villus height and villus/crypt ratio, coupled with elevated levels of intestinal inflammation and permeability. During the fasting period, microbiota compositions changed. The abundance of increased, while the abundance of and decreased. was positively correlated with Citrinin and Sterobilin, which lead to intestinal inflammation. and exhibited positive correlations with Lanthionine and reduced Glutathione, thereby reducing intestinal inflammation. This study screened the intestinal probiotics, and , that influence gut health during the fasting period, providing an experimental basis for improving gut microbiota and reducing intestinal inflammation during the FIM process.
PubMed: 38891687
DOI: 10.3390/ani14111640 -
BMC Genomics Jun 2024A growing number of studies have demonstrated that the polar regions have the potential to be a significant repository of microbial resources and a potential source of...
BACKGROUND
A growing number of studies have demonstrated that the polar regions have the potential to be a significant repository of microbial resources and a potential source of active ingredients. Genome mining strategy plays a key role in the discovery of bioactive secondary metabolites (SMs) from microorganisms. This work highlighted deciphering the biosynthetic potential of an Arctic marine-derived strain Aspergillus sydowii MNP-2 by a combination of whole genome analysis and antiSMASH as well as feature-based molecular networking (MN) in the Global Natural Products Social Molecular Networking (GNPS).
RESULTS
In this study, a high-quality whole genome sequence of an Arctic marine strain MNP-2, with a size of 34.9 Mb was successfully obtained. Its total number of genes predicted by BRAKER software was 13,218, and that of non-coding RNAs (rRNA, sRNA, snRNA, and tRNA) predicted by using INFERNAL software was 204. AntiSMASH results indicated that strain MNP-2 harbors 56 biosynthetic gene clusters (BGCs), including 18 NRPS/NRPS-like gene clusters, 10 PKS/PKS-like gene clusters, 8 terpene synthse gene clusters, 5 indole synthase gene clusters, 10 hybrid gene clusters, and 5 fungal-RiPP gene clusters. Metabolic analyses of strain MNP-2 grown on various media using GNPS networking revealed its great potential for the biosynthesis of bioactive SMs containing a variety of heterocyclic and bridge-ring structures. For example, compound G-8 exhibited a potent anti-HIV effect with an IC value of 7.2 nM and an EC value of 0.9 nM. Compound G-6 had excellent in vitro cytotoxicities against the K562, MCF-7, Hela, DU145, U1975, SGC-7901, A549, MOLT-4, and HL60 cell lines, with IC values ranging from 0.10 to 3.3 µM, and showed significant anti-viral (H1N1 and H3N2) activities with IC values of 15.9 and 30.0 µM, respectively.
CONCLUSIONS
These findings definitely improve our knowledge about the molecular biology of genus A. sydowii and would effectively unveil the biosynthetic potential of strain MNP-2 using genomics and metabolomics techniques.
Topics: Aspergillus; Arctic Regions; Humans; Multigene Family; Biological Products; Aquatic Organisms; Cell Line, Tumor; Biosynthetic Pathways; Secondary Metabolism; Genome, Fungal
PubMed: 38886660
DOI: 10.1186/s12864-024-10501-0 -
Oncology Letters Jul 2024The present study aimed to investigate the anti-leukemic effects of dihydroartemisinin (DHA) on T-cell acute lymphoblastic leukemia (T-ALL) cell lines, Jurkat and...
The present study aimed to investigate the anti-leukemic effects of dihydroartemisinin (DHA) on T-cell acute lymphoblastic leukemia (T-ALL) cell lines, Jurkat and Molt-4, and the underlying mechanisms. Cell Counting Kit-8 was performed to measure cell viability. Cell apoptosis and cell cycle distribution were assessed by flow cytometry. The expression levels of ATF4 and CHOP mRNA were assessed by reverse transcription-quantitative PCR, while the protein abundance of SLC7A11, GPX4, ATF4 and CHOP was determined by western blotting. Moreover, malondialdehyde, glutathione (GSH) and reactive oxygen species (ROS) assays were used to detect the levels of ferroptosis. The results showed that DHA suppressed T-ALL cell viability , and induced cell cycle arrest at S or G/M phase. DHA also induced ROS burst, activated endoplasmic reticulum (ER) stress, disrupted the system Xc-GSH-GSH peroxidase 4 antioxidant system, and increased lipid peroxide accumulation, resulting in cell death. By contrast, the pharmacological inhibition of ferroptosis alleviated DHA-induced cell death, confirming that DHA induces T-ALL cell death via ferroptosis. Mechanistically, the effect of DHA on ferroptosis was partly mediated by downregulating SLC7A11 and upregulating the ATF4-CHOP signaling pathway, which is associated with ER stress. These results indicated that DHA may induce ferroptosis in T-ALL cell lines and could represent a promising therapeutic agent for treating T-ALL.
PubMed: 38846431
DOI: 10.3892/ol.2024.14470 -
BMC Genomics Jun 2024Compound eyes formation in decapod crustaceans occurs after the nauplius stage. However, the key genes and regulatory mechanisms of compound eye development during... (Comparative Study)
Comparative Study
Compound eyes formation in decapod crustaceans occurs after the nauplius stage. However, the key genes and regulatory mechanisms of compound eye development during crustacean embryonic development have not yet been clarified. In this study, RNA-seq was used to investigate the gene expression profiles of Neocaridina denticulata sinensis from nauplius to zoea stage. Based on RNA-seq data analysis, the phototransduction and insect hormone biosynthesis pathways were enriched, and molting-related neuropeptides were highly expressed. There was strong cell proliferation in the embryo prior to compound eye development. The formation of the visual system and the hormonal regulation of hatching were the dominant biological events during compound eye development. The functional analysis of DEGs across all four developmental stages showed that cuticle formation, muscle growth and the establishment of immune system occurred from nauplius to zoea stage. Key genes related to eye development were discovered, including those involved in the determination and differentiation of the eye field, eye-color formation, and visual signal transduction. In conclusion, the results increase the understanding of the molecular mechanism of eye formation in crustacean embryonic stage.
Topics: Animals; Gene Expression Profiling; Compound Eye, Arthropod; Transcriptome; Gene Expression Regulation, Developmental; Decapoda; Eye
PubMed: 38844864
DOI: 10.1186/s12864-024-10453-5 -
Veterinary World Apr 2024is an obligate intraerythrocytic rickettsial parasite that infects cattle in tropical and subtropical regions. There is no evidence that inoculation can be used to...
BACKGROUND AND AIM
is an obligate intraerythrocytic rickettsial parasite that infects cattle in tropical and subtropical regions. There is no evidence that inoculation can be used to culture in rabbits. This study aimed to determine the molting of s larvae, nymphs, and adults on rabbits as well as nymphs and adults of on calves with or without . Transstadial, horizontal, and transovarial transmissions of in reared on rabbits and calves were evaluated.
MATERIALS AND METHODS
Engorged female ticks were collected from field samples of -infected and non-infected cattle. We divided the eight rabbits into two groups: A and B. Group A rabbits were infected with through parenteral inoculation, whereas Group B rabbits were kept as a control. The "clean rabbits" in Group B were observed for tick rearing without . Polymerase chain reaction was used to screen in rabbits and stages of tick. The complete life cycle of with or without was observed on rabbits.
RESULTS
A 6.5-day longer life cycle was observed in ticks harboring than in ticks without . To observe transstadial transmission, transstadial, horizontal, and transovarial transmissions of in ticks were experimentally observed in one clean calf fed separately with infected nymphs and female adult ticks.
CONCLUSION
We experimentally observed transovarian, transstadial, and transovarial transmission of in ticks as a biological vector reared on calves and rabbits. We used rabbits as a model animal for rearing ticks and culture of .
PubMed: 38798298
DOI: 10.14202/vetworld.2024.903-910 -
Molecules (Basel, Switzerland) May 2024L-asparaginases are used in the treatment of acute lymphoblastic leukemia. The aim of this work was to compare the antiproliferative potential and proapoptotic...
L-asparaginases are used in the treatment of acute lymphoblastic leukemia. The aim of this work was to compare the antiproliferative potential and proapoptotic properties of novel L-asparaginases from different structural classes, viz. EcAIII and KpAIII (class 2), as well as ReAIV and ReAV (class 3). The EcAII (class 1) enzyme served as a reference. The proapoptotic and antiproliferative effects were tested using four human leukemia cell models: MOLT-4, RAJI, THP-1, and HL-60. The antiproliferative assay with the MOLT-4 cell line indicated the inhibitory properties of all tested L-asparaginases. The results from the THP-1 cell models showed a similar antiproliferative effect in the presence of EcAII, EcAIII, and KpAIII. In the case of HL-60 cells, the inhibition of proliferation was observed in the presence of EcAII and KpAIII, whereas the proliferation of RAJI cells was inhibited only by EcAII. The results of the proapoptotic assays showed individual effects of the enzymes toward specific cell lines, suggesting a selective (time-dependent and dose-dependent) action of the tested L-asparaginases. We have, thus, demonstrated that novel L-asparaginases, with a lower substrate affinity than EcAII, also exhibit significant antileukemic properties in vitro, which makes them interesting new drug candidates for the treatment of hematological malignancies. For all enzymes, the kinetic parameters (K and k) and thermal stability (T) were determined. Structural and catalytic properties of L-asparaginases from different classes are also summarized.
Topics: Humans; Asparaginase; Antineoplastic Agents; Apoptosis; Cell Proliferation; Cell Line, Tumor; Substrate Specificity; HL-60 Cells; Leukemia
PubMed: 38792133
DOI: 10.3390/molecules29102272 -
Scientific Reports May 2024Behaviors can vary throughout an animal's life and this variation can often be explained by changes associated with learning and/or maturing. Currently, there is little...
Behaviors can vary throughout an animal's life and this variation can often be explained by changes associated with learning and/or maturing. Currently, there is little consensus regarding how these processes interact to affect behaviors. Here we proposed a heuristic approach to disentangle the effects of learning and maturation on behavior and applied it to the predatory behaviors of Physocyclus globosus spiderlings. We varied the degree of prey difficulty and familiarity spiderlings received along the first instar and across the molt to the second instar and quantified the time spiderlings spent wrapping prey, as a proxy for prey capture efficiency. We found no overall evidence for learning or maturation. Changes in efficiency were mainly due to the switch from difficult to easy prey, or vice versa. However, there was one treatment where spiderlings improved in efficiency before and after the molt, without a switch in prey type. This provides some indication that difficult prey may offer more opportunity for learning or maturation to impact behavior. Although we found little effect of learning or maturation on prey capture efficiency, we suggest that our heuristic approach is effective and could be useful in investigating these processes in other behaviors and other animals.
Topics: Animals; Spiders; Predatory Behavior; Learning; Heuristics
PubMed: 38778018
DOI: 10.1038/s41598-024-61252-7 -
BioRxiv : the Preprint Server For... May 2024Apical extracellular matrices (aECMs) are associated with all epithelia and form a protective layer against biotic and abiotic threats in the environment. Despite their...
Apical extracellular matrices (aECMs) are associated with all epithelia and form a protective layer against biotic and abiotic threats in the environment. Despite their importance, we lack a deep understanding of their structure and dynamics in development and disease. molting offers a powerful entry point to understanding developmentally programmed aECM remodeling. A transient matrix is formed in embryos and at the end of each larval stage, presumably to pattern the new cuticle. Focusing on targets of NHR-23, a key transcription factor which drives molting, we identified the Kunitz family protease inhibitor gene as an NHR-23 target. We identified NHR-23-binding sites that are necessary and sufficient for epithelial expression. is necessary to pattern every layer of the adult cuticle, suggesting a broad patterning role prior to the formation of the mature cuticle. MLT-11::mNeonGreen::3xFLAG transiently localized to the aECM in the cuticle and embryo. It was also detected in lining openings to the exterior (vulva, rectum, mouth). Reduction of function disrupted the barrier function of the cuticle. Tissue-specific RNAi suggested activity is primarily necessary in seam cells and we observed alae and seam cell fusion defects upon inactivation. Predicted null mutations caused fully penetrant embryonic lethality and elongation defects suggesting also plays an important role in patterning the embryonic sheath. Finally, we found that inactivation suppressed the blistered cuticle phenotype of mutants of mutants, a subtilisin protease gene but did not affect BLI-4::sfGFP expression. These data could suggest that MLT-11 may be necessary to assure proper levels of BLI-4 activity.
PubMed: 38766248
DOI: 10.1101/2024.05.12.593762