PI3K-Akt-SGF1-Dimm pathway mediates the nutritional regulation of silk protein synthesis in Bombyx mori.

The efficient synthesis of silk protein is heavily reliant on the ingestion of massive nutrients during the peak growth phase in the silkworm. However, the molecular mechanism of nutritional regulation of silk protein synthesis remains unknown. In this study, we investigated the impact of nutrient deficiency on the synthesis of silk protein. Nutritional deficiency led to a reduction in silk yield, accompanied by decreased levels of silk proteins and fibroin heavy chain (FibH)-activating transcription factors SGF1 and Dimm. Furthermore, insulin enhanced the protein levels of SGF1 and Dimm, which can be attenuated by specific inhibitors of PI3K. Co-immunoprecipitation analysis showed that the nutrient pathway factor protein kinase B (Akt) could interact with SGF1 protein. Knockdown of Akt reduced the phosphorylation level of SGF1 and impedes its nuclear translocation. Further studies revealed that SGF1 was directly bound to Fkh site in the 22-43 region upstream of ATG of Dimm gene to activate its transcription. In conclusion, during the peak growth phase, nutrition promotes the massive synthesis of silk protein through the PI3K-Akt-SGF1-Dimm pathway. This study offers valuable insights into the efficient synthesis of silk proteins and establishes a theoretical foundation for improving silk yield.

Silk Gland Factor 1 Plays a Pivotal Role in Larval Settlement of the Fouling Mussel Mytilopsis sallei.

Most fouling organisms have planktonic larval and benthic adult stages. Larval settlement, the planktonic-benthic transition, is the critical point when biofouling begins. However, our understanding of the molecular mechanisms of larval settlement is limited. In our previous studies, we identified that the AMP-activated protein kinase-silk gland factor 1 (AMPK-SGF1) pathway was involved in triggering the larval settlement in the fouling mussel M. sallei. In this study, to further confirm the pivotal role of SGF1, multiple targeted binding compounds of SGF1 were obtained using high-throughput virtual screening. It was found that the targeted binding compounds, such as NAD+ and atorvastatin, could significantly induce and inhibit the larval settlement, respectively. Furthermore, the qRT-PCR showed that the expression of the foot proteins' genes was significantly increased after the exposure to 10 µM NAD+, while the gene expression was significantly suppressed after the exposure to 10 µM atorvastatin. Additionally, the production of the byssus threads of the adults was significantly increased after the exposure to 10-20 µM of NAD+, while the production of the byssus threads was significantly decreased after the exposure to 10-50 µM of atorvastatin. This work will deepen our understanding of SGF1 in triggering the larval settlement in mussels and will provide insights into the potential targets for developing novel antifouling agents.

UPS writes a new saga of SAGA.

SAGA (Spt-Ada-Gcn5-Acetyltransferase), an evolutionarily conserved transcriptional co-activator among eukaryotes, is a large multi-subunit protein complex with two distinct enzymatic activities, namely HAT (Histone acetyltransferase) and DUB (De-ubiquitinase), and is targeted to the promoter by the gene-specific activator proteins for histone covalent modifications and PIC (Pre-initiation complex) formation in enhancing transcription (or gene activation). Targeting of SAGA to the gene promoter is further facilitated by the 19S RP (Regulatory particle) of the 26S proteasome (that is involved in targeted degradation of protein via ubiquitylation) in a proteolysis-independent manner. Moreover, SAGA is also recently found to be regulated by the 26S proteasome in a proteolysis-dependent manner via the ubiquitylation of its Sgf73/ataxin-7 component that is required for SAGA's integrity and DUB activity (and hence transcription), and is linked to various diseases including neurodegenerative disorders and cancer. Thus, SAGA itself and its targeting to the active gene are regulated by the UPS (Ubiquitin-proteasome system) with implications in diseases.

Splenogonadal fusion: A rare case report and literature review.

INTRODUCTION AND IMPORTANCE: Splenogonadal fusion is a rare congenital anomaly occurs when splenic tissue presents near or within a gonad. It mostly involves male children. Although it is benign and rare, making a pre-operation precise diagnosis is challenging which can lead to unnecessary invasive treatments. CASE PRESENTATION: A 3-year-old boy was presented by the chief complaint of a painless mass on the left testis and left inguinal hernia. He had a previous history of bilateral cryptorchidism and orchiopexy. Ultrasonography showed a small mass on the inferior pole of left testis and left reducible inguinal hernia. He went under left orchiectomy and hernia repair. Pathological investigation of the specimen resembled normal splenic tissue next to testicular tissue and the diagnosis of splenogonadal fusion was made. CLINICAL DISCUSSION: Splenogodal fusion cases can be challenging. Pain and sensation of mass in the scrotal sac are the most common presentation of splenogonadal fusion. Testicular malignancies can be considered as their main differential diagnosis, despite the fact that imaging and intra-operation frozen section can be helpful in making a definite diagnosis in some cases. It is mostly diagnosed incidentally during other procedures such as hernia repair or orchiopexy. Since it is benign, removal of tumor without orchiectomy is curative. CONCLUSION: In dealing with testicular mass in children, raising awareness of splenogonadal fusion have utmost importance to prevent unnecessary radical surgical interventions.

A novel ubiquitin-proteasome system regulation of Sgf73/ataxin-7 that maintains the integrity of the coactivator SAGA in orchestrating transcription.

Ataxin-7 maintains the integrity of Spt-Ada-Gcn5-Acetyltransferase (SAGA), an evolutionarily conserved coactivator in stimulating preinitiation complex (PIC) formation for transcription initiation, and thus, its upregulation or downregulation is associated with various diseases. However, it remains unknown how ataxin-7 is regulated that could provide new insights into disease pathogenesis and therapeutic interventions. Here, we show that ataxin-7's yeast homologue, Sgf73, undergoes ubiquitylation and proteasomal degradation. Impairment of such regulation increases Sgf73's abundance, which enhances recruitment of TATA box-binding protein (TBP) (that nucleates PIC formation) to the promoter but impairs transcription elongation. Further, decreased Sgf73 level reduces PIC formation and transcription. Thus, Sgf73 is fine-tuned by ubiquitin-proteasome system (UPS) in orchestrating transcription. Likewise, ataxin-7 undergoes ubiquitylation and proteasomal degradation, alteration of which changes ataxin-7's abundance that is associated with altered transcription and cellular pathologies/diseases. Collectively, our results unveil a novel UPS regulation of Sgf73/ataxin-7 for normal cellular health and implicate alteration of such regulation in diseases.

Effect of Supergravity Field on the Microstructure and Mechanical Properties of Highly Conductive Cu Alloys.

In consideration of the characteristics of supergravity to strengthen solidification structures, the effect of the supergravity field (SGF) on the grain refinement and mechanical properties of Cu-0.5Sn alloys was investigated in this paper. Firstly, it was experimentally verified that the addition of Sn could effectively refine the grain. Subsequently, the variations in grain size, tensile strength, and plasticity of the Cu-0.5Sn alloy were compared in normal and SGF conditions. The results revealed that the tensile strength and plasticity of the alloy increased with the increase in gravity coefficient. The ultimate tensile strength of the Cu-0.5Sn alloy in a normal gravity field was 145.2 MPa, while it was 160.2, 165.3, 167.9, and 182.0 MPa in an SGF with G = 100, 300, 500, and 1000, respectively, and there was almost no effect on conductivity. Finally, it was clarified that the mechanism of grain refinement by SGF was that the intense convection caused the fracture of the dendrites to become new nucleating particles. The increased viscosity under SGF hindered the diffusion of atoms in the melt and slowed down the movement of atoms toward the nucleus, leading to a decrease in grain size.

Novel cationic cellulose beads for oral delivery of poorly water-soluble drugs.

Cellulose beads emerge as carriers for poorly water-soluble drugs due to their eco-friendly raw materials and favorable porous structure. However, drug dissolution may be limited by their poor swelling ability and the presence of closed pores caused by shrinkage of the pristine cellulose beads. In this study, novel cellulose beads that can swell in acidic environment were prepared by introducing ethylenediamine (EDA) on dialdehyde cellulose (DAC), thereby addressing the shrinkage and closed pore problem of cellulose beads. The effect of the ratio of EDA on the swelling behavior and amine content of beads was studied. Three model drugs with different physicochemical properties were selected to study the physical state of loaded drugs and their release behavior. According to the results of XRPD and DSC, indomethacin and itraconazole loaded in the beads were amorphous at a drug loading of 20%, but fenofibrate was partially crystalline. Both bead size and the ratio of amine groups influenced the release behavior of the model drugs. The in vitro dissolution results showed that the cationic beads greatly improved the solubility and dissolution rate of the drug compared with the crystalline drug. Beads with a small size and high ratio of EDA tend to achieve a better drug dissolution rate and cumulative release percentage. Physical stability studies of the itraconazole-loaded beads were also implemented under four different temperature/humidity conditions for up to two months. The results showed that crystallization only appeared after two months of storage at 40°/75% RH, and the drug maintained a non-crystalline state in the other three storage conditions (0 °C/0 %RH, 0 °C/32 %RH, 25 °C/32 %RH). In conclusion, the novel pH-responsive cationic cellulose beads show great potential as a carrier for improving the rate and extent of dissolution of poorly soluble drugs and maintaining supersaturation.

TB500/TB1000 and SGF1000: A scientific approach for a better understanding of misbranded and adulterated drugs.

Nowadays, numerous websites attempt to commercialize over the internet various products, regardless of the lack of approval by the EMA or the FDA either for human or veterinary use. These products are often produced after aborted drug development due to insufficient or deleterious biological effects, synthesized based on natural products, or only based on scientific literature. However, the administration of such products is dangerous, considering the lack of official control over the production of these substances and the absence of approval by health authorities. In this short communication, we provide an extensive analysis of three misbranded and adulterated products sold over the internet named TB500, TB1000, and SGF1000. We confirm that the content of TB500/TB1000 products is not systematically consistent with it's former descriptions, but also that SGF1000 is mainly composed of sheep extracellular matrix (ECM) and blood proteins, and the signal corresponding to the purported growth promoters is excessively diluted.

A Novel Framework for Analysis of the Shared Genetic Background of Correlated Traits.

We propose a novel effective framework for the analysis of the shared genetic background for a set of genetically correlated traits using SNP-level GWAS summary statistics. This framework called SHAHER is based on the construction of a linear combination of traits by maximizing the proportion of its genetic variance explained by the shared genetic factors. SHAHER requires only full GWAS summary statistics and matrices of genetic and phenotypic correlations between traits as inputs. Our framework allows both shared and unshared genetic factors to be effectively analyzed. We tested our framework using simulation studies, compared it with previous developments, and assessed its performance using three real datasets: anthropometric traits, psychiatric conditions and lipid concentrations. SHAHER is versatile and applicable to summary statistics from GWASs with arbitrary sample sizes and sample overlaps, allows for the incorporation of different GWAS models (Cox, linear and logistic), and is computationally fast.

Viability of Lactobacillus delbrueckii in chocolates during storage and in-vitro bioaccessibility of polyphenols and SCFAs.

This study evaluated the viability of encapsulated Lactobacillus delbrueckii subsp. bulgaricus in chocolate during storage and in-vitro gastrointestinal transit. Flavonoid contents and short chain fatty acids (SCFAs) production during gastrointestinal transit were also assessed. Encapsulated L. delbrueckii subsp. bulgaricus survived well in chocolates >7 logs both after 120 days of storage at 4 °C and 25 °C, and during in-vitro gastrointestinal transit. The release of SCFAs through in-vitro gastrointestinal digestion and colonic fermentation revealed that probiotic-chocolates could be an excellent source of nutrients for the gut microbiota. Encapsulated probiotic in chocolates with 70% cocoa produced significantly (P < 0.05) more acetic, propionic, isobutyric, butyric and isovaleric acids than that with 45% cocoa. The bioconversion results of a specific polyphenol by L. delbrueckii subsp. bulgaricus exhibited that chocolate polyphenols could be utilized by probiotics for their metabolism. These findings confirmed that chocolate could be successfully fortified with L. delbrueckii subsp. bulgaricus encapsulation to improve health promoting properties of chocolates.

Chiral mesoporous silica nano-screws as an efficient biomimetic oral drug delivery platform through multiple topological mechanisms.

In the microscale, bacteria with helical body shapes have been reported to yield advantages in many bio-processes. In the human society, there are also wisdoms in knowing how to recognize and make use of helical shapes with multi-functionality. Herein, we designed atypical chiral mesoporous silica nano-screws (CMSWs) with ideal topological structures (e.g., small section area, relative rough surface, screw-like body with three-dimension chirality) and demonstrated that CMSWs displayed enhanced bio-adhesion, mucus-penetration and cellular uptake (contributed by the macropinocytosis and caveolae-mediated endocytosis pathways) abilities compared to the chiral mesoporous silica nanospheres (CMSSs) and chiral mesoporous silica nanorods (CMSRs), achieving extended retention duration in the gastrointestinal (GI) tract and superior adsorption in the blood circulation (up to 2.61- and 5.65-times in AUC). After doxorubicin (DOX) loading into CMSs, DOX@CMSWs exhibited controlled drug release manners with pH responsiveness in vitro. Orally administered DOX@CMSWs could efficiently overcome the intestinal epithelium barrier (IEB), and resulted in satisfactory oral bioavailability of DOX (up to 348%). CMSWs were also proved to exhibit good biocompatibility and unique biodegradability. These findings displayed superior ability of CMSWs in crossing IEB through multiple topological mechanisms and would provide useful information on the rational design of nano-drug delivery systems.

Isolation and characterization of SGF3, a novel Microviridae phage infecting Shigella flexneri.

In the context of widespread bacterial contamination and the endless emergence of antibiotic-resistant bacteria, more effective ways to control pathogen infection are urgently needed. Phages become potential bactericidal agents due to their bactericidal specificity and not easy resistance to bacteria. But an important factor limiting its development is the lack of phage species. Therefore, the isolation of more new phages and studying their biological and genomic characteristics is of great significance for subsequent applications. So, in this study, SGF3, a Microviridae phage, which has shown lytic activity against Shigella flexneri, was isolated, purified, and characterized. Morphological and phylogenetic analyses identified it as a phiX174 species belonging to the Microviridae family. The latent period of phage SGF3 was 20 min, with an average burst size of approximately 7.1. Host spectrum experiments indicated its strong host specificity. Furthermore, the biofilm removal efficiency was increased by 20%-25% when SGF3 was coupled with other phages. In conclusion, the phage SGF3 found in this study was a lytic phage belonging to the Microviral family, and could be added as an auxiliary material in the phage cocktail. Studies of its characteristics and bactericidal properties had enriched the germplasm resources of microphages, provided more potential material in fighting against emerging and existing multidrug-resistant bacteria.

Oral delivery of proteins and peptides: Challenges, status quo and future perspectives.

Proteins and peptides (PPs) have gradually become more attractive therapeutic molecules than small molecular drugs due to their high selectivity and efficacy, but fewer side effects. Owing to the poor stability and limited permeability through gastrointestinal (GI) tract and epithelia, the therapeutic PPs are usually administered by parenteral route. Given the big demand for oral administration in clinical use, a variety of researches focused on developing new technologies to overcome GI barriers of PPs, such as enteric coating, enzyme inhibitors, permeation enhancers, nanoparticles, as well as intestinal microdevices. Some new technologies have been developed under clinical trials and even on the market. This review summarizes the history, the physiological barriers and the overcoming approaches, current clinical and preclinical technologies, and future prospects of oral delivery of PPs.

In vitro and in vivo correlation for lipid-based formulations: Current status and future perspectives.

Lipid-based formulations (LBFs) have demonstrated a great potential in enhancing the oral absorption of poorly water-soluble drugs. However, construction of in vitro and in vivo correlations (IVIVCs) for LBFs is quite challenging, owing to a complex in vivo processing of these formulations. In this paper, we start with a brief introduction on the gastrointestinal digestion of lipid/LBFs and its relation to enhanced oral drug absorption; based on the concept of IVIVCs, the current status of in vitro models to establish IVIVCs for LBFs is reviewed, while future perspectives in this field are discussed. In vitro tests, which facilitate the understanding and prediction of the in vivo performance of solid dosage forms, frequently fail to mimic the in vivo processing of LBFs, leading to inconsistent results. In vitro digestion models, which more closely simulate gastrointestinal physiology, are a more promising option. Despite some successes in IVIVC modeling, the accuracy and consistency of these models are yet to be validated, particularly for human data. A reliable IVIVC model can not only reduce the risk, time, and cost of formulation development but can also contribute to the formulation design and optimization, thus promoting the clinical translation of LBFs.

The Histone Acetyltransferase GCN5 and the Associated Coactivators ADA2: From Evolution of the SAGA Complex to the Biological Roles in Plants.

Transcription of protein-encoding genes starts with forming a pre-initiation complex comprised of RNA polymerase II and several general transcription factors. To activate gene expression, transcription factors must overcome repressive chromatin structure, which is accomplished with multiprotein complexes. One such complex, SAGA, modifies the nucleosomal histones through acetylation and other histone modifications. A prototypical histone acetyltransferase (HAT) known as general control non-repressed protein 5 (GCN5), was defined biochemically as the first transcription-linked HAT with specificity for histone H3 lysine 14. In this review, we analyze the components of the putative plant SAGA complex during plant evolution, and current knowledge on the biological role of the key components of the HAT module, GCN5 and ADA2b in plants, will be summarized.

Splenogonadal fusion: a case report and review of the literature.

BACKGROUND: Splenogondal fusion (SGF) is a rare congenital anomaly characterized by abnormal association between the splenic tissue and the gonads or mesonephric remnants. SGF that requires separate two-stage laparoscopic staged Fowler-Stephen orchiopexy on both the left and right sides is extremely rare. SGF could be misdiagnosed as testicular malignancy and leads to unnecessary orchiectomy. CASE PRESENTATION: This is a case of an 8-month old male infant presented with bilateral cryptorchidism, B-mode ultrasound visualized the left and right testes in the lower abdominal cavity and the upper margin of the left testicle as a hypoechoic mass extending to the spleen, indicating an undescended right testis and possible SGF on the left side. Single-site laparoscopic examination confirmed the diagnosis of SGF on the left side and an undescended right testis. As both testes were high and the right spermatic vessel was poorly developed and short, a routine single stage orchiopexy would be difficult and risky, therefore, separate two-stage laparoscopic staged Fowler-Stephen orchiopexies for both sides were implemented. Stage 1 of the staged Fowler-Stephen orchiopexy for the right side was performed first without treating the left side, Stage 2 for the right side, separation of the left testis from the spleen as well as Stage 1 for the left side were performed 7 months later, and Stage 2 for the left side was performed 7 months after that. Follow-up ultrasound 1 year after the surgery revealed no obvious abnormalities in the shapes of the testes or their blood supply. This treatment strategy prevented unnecessary orchiectomy. CONCLUSIONS: We reported a rare case of SGF that needed separate two-stage laparoscopic staged Fowler-Stephen orchiopexies for both sides, and a review of the recent literature. SGF is a rare congenital anomaly often diagnosed incidentally during exploration/surgery for scrotal swelling/mass, cryptorchidism or inguinal hernia in young patients. Surgeons, especially pediatric surgeons should be aware of this rare condition to avoid unnecessary, life-altering radical orchiectomy. When routine single stage orchiopexy is not feasible or risky for either side, separate two-stage laparoscopic staged Fowler-Stephen orchiopexies could be performed on both the left and right sides to avoid unnecessary orchiectomy.

Splenogonadal fusion: A rare case report and literature review.

Splenogonadal fusion is a rare benign congenital anomaly with few cases described in the literature. It is 16 times more common in males than in females. A 22 year-old healthy male with cryptorchidism presented with preoperative imaging strongly suggestive of malignancy. Histopathology after left orchiectomy showed mixed splenic and testicular tissue with no sign of malignancy. Splenogonadal fusion is rarely diagnosed preoperatively. It should be included in differential diagnoses in patients presenting with a testicular or abdominal mass. Greater recognition of this rare anomaly may facilitate testis sparing surgery in future cases.

Fates of Au, Ag, ZnO, and CeO2 Nanoparticles in Simulated Gastric Fluid Studied using Single-Particle-Inductively Coupled Plasma-Mass Spectrometry.

The increasing use of engineered nanoparticles (ENPs) in many industries has generated significant research interest regarding their impact on the environment and human health. The major routes of ENPs to enter the human body are inhalation, skin contact, and ingestion. Following ingestion, ENPs have a long contact time in the human stomach. Hence, it is essential to know the fate of the ENPs under gastric conditions. This study aims to investigate the fate of the widely used nanoparticles Ag-NP, Au-NP, CeO2-NP, and ZnO-NP in simulated gastric fluid (SGF) under different conditions through the application of single-particle inductively coupled plasma-mass spectrometry (SP-ICP-MS). The resulting analytical methods have size detection limits for Ag-NP, Au-NP, ZnO-NP, and CeO2-NP from 15 to 35 nm, and the particle concentration detection limit is 135 particles/mL. Metal ions corresponding to the ENPs of interest were detected simultaneously with detection limits from 0.02 to 0.1 µg/L. The results showed that ZnO-NPs dissolved completely and rapidly in SGF, whereas Au-NPs and CeO2-NPs showed apparent aggregation and did not dissolve significantly. Both aggregation and dissolution were observed in Ag-NP samples following exposure to SGF. The size distributions and concentrations of ENPs were affected by the original ENP concentration, ENP size, the contact time in SGF, and temperature. This work represents a significant advancement in the understanding of ENP characteristics under gastric conditions.

[Digestive stability in simulated gastric fluid of EPSPS protein and PAT protein].

OBJECTIVE: To study the digestive stability of 5-enolpyruvylshikimate-3-phosphate synthase(EPSPS) protein and phosphinothricina cetyltransferase(PAT) protein in simulated gastric fluid. METHODS: The component of simulated gastric fluid was based on the method of target protein digestive stability in simulative gastric and intestinal in national standard of the People's Republic of China(Published by the Ministry of Agriculture No. 869-2-2007). The test model of stability of different protein to digestion in Simulated Gastric Fluid was established by dodecyl sulfate, sodium salt-polyacrylamide gel electrophoresis(SDS-PAGE)and western blot. The degradation of EPSPS protein and PAT protein in simulated gastric fluid at different digestion time points were analyzed. RESULTS: The experiment showed that EPSPS protein and PAT protein were completely digested within 15 s in simulated gastric fluid, no any remain of protein was detected by SDS-PAGE and Western blot, indicating that EPSPS protein and PAT protein were easily digested in the simulated gastric. CONCLUSION: EPSPS protein and PAT protein do not have immunogenicity after digestion with simulated gastric fluid.

Characterization of the soybean R2R3-MYB transcription factor GmMYB81 and its functional roles under abiotic stresses.

R2R3-type MYBs are a key group of regulatory factors that control diverse developmental processes and stress tolerance in plants. Soybean is a major legume crop with the richness of seed protein and edible vegetable oil, and 244 R2R3-type MYBs have been identified in soybean. However, the knowledge regarding their functional roles has been greatly limited as yet. In this study, a novel R2R3-type MYB (GmMYB81) was functionally characterized in soybean, and it is closely related to two abiotic stress-associated regulators (AtMYB44 and AtMYB77). GmMYB81 transcripts not only differentially accumulated in soybean tissues and during embryo development, but also were significantly enhanced by drought, salt and cold stress. Histochemical GUS assay in Arabidopsis indicated that GmMYB81 promoter showed high activity in seedlings, rosette leaves, inflorescences, silique wall, mature anthers, roots, and germinating seeds. Further investigation indicated that over-expression of GmMYB81 in Arabidopsis caused auxin-associated phenotypes, including small flower and silique, more branch, and weakened apical dominance. Moreover, over-expression of GmMYB81 significantly elevated the rates of seed germination and green seedling under salt and drought stress, indicating that GmMYB81 might confer plant tolerance to salt and drought stress during seed germination. Additionally, protein interaction analysis showed that GmMYB81 interacts with the abiotic stress regulator GmSGF14l. Further observation indicated that they displayed similar expression patterns under drought and salt stress, suggesting GmMYB81 and GmSGF14l might cooperatively affect stress tolerance. These findings will facilitate future investigations of the regulatory mechanisms of GmMYB81 in response to plant stress tolerance, especially seed germination under abiotic stresses.