It has been proposed that prohibitin (PHB) involved in multiple cellular functions, including cell proliferation, differentiation, apoptosis, senescence and carcinogenesis. Various cellular ...compartment location of PHB demonstrates its diverse roles. Based on the full-length sequence of PHB gene, we analyzed the deduced amino acid sequence and the predicted protein structure of this gene in the swimming crab Charybdis japonica. It shows that the structure and function of PHB are conservative. The expression level of PHB mRNA and protein in different tissues were analyzed by sqRT-PCR and western blot respectively, which showed its high expression in testis. We then traced PHB protein by immunofluorescence, and we found its diverse distribution in cytoplasm and mitochondria at different stages. We propose that PHB may participate actively in spermatogenic cell anti-apoptosis, cell nucleus distortion as well as acrosome morphogenesis during the spermatogenesis in Charybdis japonica. Furthermore, PHB was found to be ubiquitinated at different levels. Its signal was weak in spermatocytes and Stage 1 spermatids, stronger in stage 2–4 spermatids, and lowest in mature sperm. Our data shows that PHB may mediate the paternal mitochondrial material degradation by ubiquitination. We conclude that PHB is indispensable in the spermatogenesis of the swimming crab Charybdis japonica through different testis developmental stages.
•The characteristics of C. japonica PHB gene and protein were investigated.•The sub-cellular location of PHB was confirmed in C. japonica germ cells.•The ubiquitination state of PHB was explored during C. japonica spermatogenesis.
Phthalates are widely used synthetic chemicals that determine endocrine disruption effects on female reproductivity and oviposition. Our study demonstrated that the mitochondrial quality in ovarian ...granulosa cells (GCs) is associated with a poor prognosis in female reproduction. However, the molecular mechanism of di-(2-ethylhexyl) phthalate (DEHP) exposure on the quail ovarian GC layer is still unknown. To validate the effects of DEHP on the GC layer, 8 days’ old 150 female Japanese quail were treated orally with DEHP (250, 500, and 750 mg/kg BW/day) for 45 days to explore the toxic effects of DEHP on the ovarian GC layer. Histopathological assessment and ultrastructure observation found that DEHP decreased the thickness of the GC layer, resulted in mitochondrial damage, and activated mitocytosis. Additionally, the results further suggested that DEHP impacted the secretion of steroid hormones (reduced FSH, E2, and T levels and boosted Prog, PRL, and LH levels) by triggering mitocytosis (enhanced transcription of MYO19 and protein of KIF5B levels), mitochondrial dynamics (increasing mRNA and protein levels of OPA1, DRP1, MFN1, and MFN2), mitophagy (increasing mRNA and protein levels of Parkin, LC3B, and P62), and inducing GC function disorder. In conclusion, our research provided a new idea to explain the mechanism of DEHP toxicity of the ovarian GC layer in quail and presented insights into the role of mitocytosis in DEHP-induced ovarian GC layer injury.
Francisella tularensis
is a dangerous pathogen that causes an extremely contagious zoonosis in humans named tularemia. Given its low-dose morbidity, the potential to be fatal, and aerosol spread, it ...is regarded as a severe threat to public health. The US Centers for Disease Control and Prevention (CDC) has classified it as a category A potential agent for bioterrorism and a Tier 1 Select Agent. Herein, we combined recombinase polymerase amplification (RPA) with CRISPR/Cas12a system to select the
F. tularensis
target gene (TUL4), creating a two-pronged rapid and ultrasensitive diagnostic method for detecting
F. tularensis
. The real-time RPA (RT-RPA) assay detected
F. tularensis
within 10 min at a sensitivity of 5 copies/reaction,
F. tularensis
genomic DNA of 5 fg, and
F. tularensis
of 2 × 10
2
CFU/ml; the RPA-CRISPR/Cas12a assay detects
F. tularensis
within 40 min at a sensitivity of 0.5 copies/reaction,
F. tularensis
genomic DNA of 1 fg, and
F. tularensis
of 2 CFU/ml. Furthermore, the evaluation of specificity showed that both assays were highly specific to
F. tularensis
. More importantly, in a test of prepared simulated blood and sewage samples, the RT-RPA assay results were consistent with RT-PCR assay results, and the RPA-CRISPR/Cas12a assay could detect a minute amount of
F. tularensis
genomic DNA (2.5 fg). There was no nonspecific detection with blood samples and sewage samples, giving the tests a high practical application value. For example, in on-site and epidemic areas, the RT-RPA was used for rapid screening and the RPA-CRISPR/Cas12a assay was used for more accurate diagnosis.
Amyloid-β (Aβ) plays an important role in the neuropathology of Alzheimer’s disease (AD), but some factors promoting Aβ generation and Aβ oligomer (Aβo) neurotoxicity remain unclear. We here find ...that the levels of ArhGAP11A, a Ras homology GTPase-activating protein, significantly increase in patients with AD and amyloid precursor protein (APP)/presenilin-1 (PS1) mice. Reducing the ArhGAP11A level in neurons not only inhibits Aβ generation by decreasing the expression of APP, PS1, and β-secretase (BACE1) through the RhoA/ROCK/Erk signaling pathway but also reduces Aβo neurotoxicity by decreasing the expressions of apoptosis-related p53 target genes. In APP/PS1 mice, specific reduction of the ArhGAP11A level in neurons significantly reduces Aβ production and plaque deposition and ameliorates neuronal damage, neuroinflammation, and cognitive deficits. Moreover, Aβos enhance ArhGAP11A expression in neurons by activating E2F1, which thus forms a deleterious cycle. Our results demonstrate that ArhGAP11A may be involved in AD pathogenesis and that decreasing ArhGAP11A expression may be a promising therapeutic strategy for AD treatment.
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•The levels of ArhGAP11A are significantly increased in patients with AD and APP/PS1 mice•ArhGAP11A increases Aβ production through RhoA/ROCK/Erk signaling pathway•ArhGAP11A promotes Aβo neurotoxicity by p53-mediated apoptosis pathway•Aβos activate the transcription factor E2F1 and elevate ArhGAP11A expression
Huang et al. show that the levels of ArhGAP11A are significantly increased in patients with AD and APP/PS1 mice. ArhGAP11A participates in the neuropathological development of AD by enhancing Aβ generation and Aβo neurotoxicity. Specific reduction of ArhGAP11A expression in neurons significantly improves the cognitive deficits of APP/PS1 mice.
High-mobility group (HMG)-box proteins are involved in chromatin organization in eukaryotes, especially in sex determination and regulation of mitochondrial DNA compaction. Although a novel HMG-box ...protein, PoxHmbB, had been initially identified to be required for filter paper cellulase activity by
Penicillium oxalicum
, the biological roles of HMG-box proteins in biomass-degrading enzyme production have not been systematically explored. The
P. oxalicum
mutant ∆
PoxHmbB
lost 34.7–86.5% of cellulase (endoglucanase,
p
-nitrophenyl-β-cellobiosidase, and
p
-nitrophenyl-β-glucopyranosidase) activities and 60.3% of xylanase activity following Avicel induction, whereas it exhibited about onefold increase in amylase activity following soluble corn starch induction. Furthermore, ∆
PoxHmbB
presented delayed conidiation and hyphae growth. Transcriptomic profiling and real-time quantitative reverse transcription-PCR revealed that
PoxHmbB
regulated the expression of major genes encoding plant biomass-degrading enzymes such as
PoxCel7A-2
,
PoxCel5B
,
PoxBgl3A
,
PoxXyn11B
, and
PoxGA15A
, as well as those involved in conidiation such as
PoxBrlA
. In vitro binding experiments further confirmed that PoxHmbB directly binds to the promoter regions of these major genes. These results further indicate the diversity of the biological functions of HMG-box proteins and provide a novel and promising engineering target for improving plant biomass-degrading enzyme production in filamentous fungi.
The effects of water addition and noodle thickness on the surface tackiness of frozen cooked noodles (FCNs) were investigated. The results showed that the FCNs with water addition of 34% had high ...disulfide group content and small ice crystal in the freezing process, which was a benefit for forming a stable gluten network. The texture characteristics (hardness, chewiness, and tensile strength) were at the best level and the surface tackiness exhibited the lowest level. Besides, the low moisture area in the recooked FCNs increased with the ascent of noodle thickness and the hardness of thick FCNs was high for forming reliable spatial support. The texture characteristics of recooked FCNs (thickness 2 mm) were better. Combined with the sensory evaluation, when the water addition was 34% and the thickness was 2 mm, the surface tackiness of recooked FCNs was lower, and the FCNs had a good quality and taste.
Practical applications
In this study, the effects of water addition and noodle thickness on the surface tackiness of frozen cooked noodles (FCNs) were investigated. When the water addition (34%) and noodle thickness (2 mm) were at the appropriate level, the surface tackiness of recooked FCNs was low, and the FCNs had a good taste and appearance. The texture properties and sensory evaluation of the FCNs were the best. The study provides a reference for processing the FCNs with low surface tackiness in the fast‐food industry. At present, the FCNs made according to this research results are widely used in the Chinese mainland market. The water addition of FCNs is 34%~36%, and the shape of FCNs is basically 20 cm length, 3 mm width, and 2 mm thickness.
Hormone therapy (HT) has failed to improve learning and memory in postmenopausal women according to recent clinical studies; however, the reason for failure of HT in improving cognitive performance ...is unknown. In our research, we found cognitive flexibility was improved by 17β-Estradiol (E2) in mice 1 week after ovariectomy (OVXST), but not in mice 3 months after ovariectomy (OVXLT). Isobaric tags for relative and absolute quantitation (iTRAQ) revealed increased cannabinoid receptor interacting protein 1 (CNRIP1) in E2-treated OVXLT mice compared with E2-treated OVXST mice. Adeno-associated virus 2/9 (AAV2/9) delivery of Cnrip1 short-hairpin small interfering RNA (Cnrip1-shRNA) rescued the impaired cognitive flexibility in E2 treated OVXLT mice. This effect is dependent on CB1 function, which could be blocked by AM251—a CB1 antagonist. Our results indicated a new method to increasing cognitive flexibility in women receiving HT by disrupting CNRIP1.
•Estrogen fails to rescue cognitive flexibility in long-term estrogen-deprived female mice.•CNRIP1 protein is increased in long-term estrogen-deprived female mice compared with short-term group.•Disrupting CNRIP1 rescues cognitive flexibility long-term estrogen-deprived female mice after estrogen treatment.
Reported herein is an intramolecular dehydrogenative coupling of two inert aryl C–H bonds for the synthesis of aporphine analogues. The process represents a novel tool for the preparation of ...aporphines via palladiun-catalyzed C–H bond activation. The present reaction is compatible with various functional groups, and the coupling products have been further applied for the synthesis of natural products aporphine and zenkerine.
This work aimed to develop a novel colorimetric indicator film for monitoring of food freshness based on gelatin/polyvinyl alcohol matrix incorporated with anthocyanin extracts from mulberry. The ...color of anthocyanin extracts solutions obviously changed from bright red to dark green in the pH range of 2.0–11.0. FTIR spectra and isothermal titration calorimetry showed that the anthocyanin extracts were successfully combined with gelatin/polyvinyl alcohol matrix by hydrogen binding and electrostatic interaction, which enhanced the stability of anthocyanin. The scanning electric microscopy showed that the compatibility between polyvinyl alcohol and gelatin were improved owing to the addition of anthocyanin extracts. With the anthocyanin extracts addition from 0 to 45 mg/100 mL mixed solution, the tensile strength decreased from 30.80 to 21.01 MPa and the elongation at break increased from 589.22% to 905.86%. The color response of film in buffer solution of different pH were in accordance with anthocyanin extracts solutions, and its color changes were clearly visible with naked eye. Finally, the film was evaluated by a test on monitoring fish spoilage, which presented visible color changes due to volatile nitrogenous compounds formed over time. These results showed that this developed film could be used as an effective method for the monitoring of food freshness.
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•A colorimetric film was developed by using gelatin, PVA and mulberry anthocyanin.•The anthocyanin stability was enhanced by combining with film matrix.•The mechanical properties of films were improved due to the addition of anthocyanin.•Color changes of film in buffer solution of different pH were visible with naked eye.•The film could be used as an effective method for the monitoring of fish spoilage.
