Molecular Logic Gate for Sensing pH/Peroxynitrite with Potential Applications in Cisplatin Treatment.

Cisplatin is a common chemotherapy drug for multiple solid tumors; however, due to the nitrification of peroxynitrite (ONOO-), a series of side effects seriously affect its dose and efficacy. Considering that the reactivity of ONOO- is significantly affected by pH in vitro, revealing their roles in living cells contributes to understanding the side-effect process induced by cisplatin. Herein, we present a near-infrared fluorescent logic gate for sensing pH/ONOO-, which can accurately discriminate four scenarios (no analyte, analyte H+ alone, analyte ONOO- alone, and H+ + ONOO-) and which one comes first. With this probe, the significant roles of pH and ONOO- in cisplatin treatment are disclosed, in which the cell account shows a dramatic reduction accompanied by decreased pH and upregulated ONOO- levels. By artificially recovering the pH, the ONOO- content and cell account can maintain a stable state, possibly due to the protection from acidification and nitration. This work provides an ideal pH/ONOO- logical sensor for revealing their potential roles under cisplatin, which is expected to proffer new insights into more related diseases and drug research.

POI-associated EIF4ENIF1 mutations exhibit impaired translation regulation abilities.

Various genetic variants have been found to be associated with the clinical onset of premature ovarian insufficiency (POI). However, when measured in vitro, the functional influence of the variants can be difficult to determine. By whole-exome sequencing (WES) of 93 patients with sporadic POI, we found a missense variant c.623G > A;p.R208H in the EIF4ENIF1 gene. In silico prediction of the variant using different algorithms suggested it might be a damaging variant. We compared the property of EIF4ENIF1 R208H and Q842P, a POI-related mutant that we reported previously, with wildtype (WT) protein using 293FT cells in vitro. Surprisingly, a change in subcellular distribution and granule forming ability (Q842P) and nuclear import capacity (R208H) was not observed, despite domain prediction evidences. Since EIF4ENIF1 was reported to inhibit translation, we employed T&T-seq, a translation-transcription dual-omics sequencing method, to profile gene expression upon overexpression of EIF4ENIF1 WT and mutants. EIF4ENIF1 WT overexpression group exhibited significantly (P < 0.0001) lower translation efficiency (TE) than empty vector or GFP overexpression control group. Surprisingly, EIF4ENIF1 Q842P overexpression failed to repress global translation, showing an overall TE significantly higher than WT group. Overexpression R208H significantly (P < 0.0001) lowered the overall TE, whereas exhibiting a reduced translation inhibitory effect on high-TE genes (TE > 2 in GFP control group). Several fertility-associated genes, such as AMH in Q842P group and SERPINE1 and THBS1 in R208H group, was translationally up-regulated in mutant groups versus WT control, suggesting a potential mechanism of mutated EIF4ENIF1 causing POI via impaired translation repression. It is further proposed that T&T-seq can be a sensitive evaluation tool for the measurement of functional alteration by variants in many other translational regulator genes, not only EIF4ENIF1, helping to eliminate misinterpretation of clinical significance of genetic variants.

FOXL2 and NR5A1 induce human fibroblasts into steroidogenic ovarian granulosa-like cells.

Human granulosa cells in different stages are essential for maintaining normal ovarian function, and granulosa cell defect is the main cause of ovarian dysfunction. To address this problem, it is necessary to induce functional granulosa cells at different stages in vitro. In this study, we established a reprogramming method to induce early- and late-stage granulosa cells with different steroidogenic abilities. We used an AMH-fluorescence-reporter system to screen candidate factors for cellular reprogramming and generated human induced granulosa-like cells (hiGC) by overexpressing FOXL2 and NR5A1. AMH-EGFP+ hiGC resembled human cumulus cells in transcriptome profiling and secreted high levels of oestrogen and progesterone, similar to late-stage granulosa cells at antral or preovulatory stage. Moreover, we identified CD55 as a cell surface marker that can be used to isolate early-stage granulosa cells. CD55+ AMH-EGFP- hiGC secreted high levels of oestrogen but low levels of progesterone, and their transcriptome profiles were more similar to early-stage granulosa cells. More importantly, CD55+ hiGC transplantation alleviated polycystic ovary syndrome (PCOS) in a mouse model. Therefore, hiGC provides a cellular model to study the developmental program of human granulosa cells and has potential to treat PCOS.

Effects of Thalidomide on Erythropoiesis and Iron Homeostasis in Transfusion-Dependent ß-Thalassemia.

Thalidomide is a therapeutic option for patients with ß-thalassemia by increasing fetal hemoglobin and thereby reducing the requirement for blood transfusions. However, information on changes in erythropoiesis and iron homeostasis during thalidomide treatment is lacking. This study investigated the effects of thalidomide treatment on hematologic, erythropoietic, and ironstatus parameters in 22 patients with transfusion-dependent ß-thalassemia (TDT). Thalidomide significantly improved anemia endpoints, including increases in hemoglobin (p<0.001), red blood cells (p<0.001), and hematocrit (p<0.001), as well as reducing erythropoietin levels (p=0.033) and ameliorating erythropoiesis. Thalidomide treatment significantly reduced serum iron levels (p=0.018) and transferrin saturation (p=0.039) and increased serum transferrin levels (p=0.030). Thalidomide had no observed effect on serum ferritin or hepcidin, but changes in hepcidin (r=0.439, p=0.041) and serum iron (r=-0.536, p=0.010) were significantly correlated with hemoglobin increment. This comprehensive study indicates that thalidomide treatment can ameliorate erythropoiesis and iron homeostasis in patients with TDT, thus supporting the effectiveness of this drug.

Eif4enif1 haploinsufficiency disrupts oocyte mitochondrial dynamics and leads to subfertility.

Infertility affects couples worldwide. Premature ovarian insufficiency (POI) refers to loss of ovarian function before 40 years of age and is a contributing factor to infertility. Several case studies have reported dominant-inherited POI symptoms in families with heterozygous EIF4ENIF1 (4E-T) mutations. However, the effects of EIF4ENIF1 haploinsufficiency have rarely been studied in animal models to reveal the underlying molecular changes related to infertility. Here, we demonstrate that Eif4enif1 haploinsufficiency causes mouse subfertility, impairs oocyte maturation and partially arrests early embryonic development. Using dual-omic sequencing, we observed that Eif4enif1 haploinsufficiency significantly altered both transcriptome and translatome in mouse oocytes, by which we further revealed oocyte mitochondrial hyperfusion and mitochondria-associated ribonucleoprotein domain distribution alteration in Eif4enif1-deficient oocytes. This study provides new insights into the molecular mechanisms underlying clinical fertility failure and new avenues to pursue new therapeutic targets to address infertility.

THRSP identified as a potential hepatocellular carcinoma marker by integrated bioinformatics analysis and experimental validation.

Hepatocellular carcinoma (HCC) is the most common malignant liver tumor with high mortality and poor prognosis worldwide. This study aimed to identify hub genes and investigate the underlying molecular mechanisms in HCC progression by integrated bioinformatics analysis and experimental validation. Based on the Gene Expression Omnibus (GEO) databases and The Cancer Genome Atlas (TCGA), 12 critical differential co-expression genes were identified between tumor and normal tissues. Via survival analysis, we found higher expression of LCAT, ACSM3, IGF1, SRD5A2, THRSP and ACADS was associated with better prognoses in HCC patients. Among which, THRSP was selected for the next investigations. We found that THRSP mRNA expression was negatively correlated with its methylation and closely associated with clinical characteristics in HCC patients. Moreover, THRSP expression had a negative correlation with the infiltration levels of several immune cells (e.g., B cells and CD4+ T cells). qRT-PCR verified that THRSP was lower expressed in HCC tissues and cell lines compared with control. Silencing of THRSP promoted the migration, invasion, proliferation, and inhibited cell apoptosis of HCCLM and Huh7 cell lines. Decreased expression of THRSP promoted HCC progression by NF-κB, ERK1/2, and p38 MAPK signaling pathways. In conclusion, THRSP might serve as a novel biomarker and therapeutic target of HCC.

Hydraulically Coupled Dielectric Elastomer Actuators for a Bioinspired Suction Cup.

Suction cups of cephalopods show a preeminent performance when absorbing irregular or flat objects. In this paper, an octopi-inspired suction cup, driven by hydraulically coupled dielectric elastomer actuators (HCDEAs), is proposed, which is considered to be controlled easily and have compact structure. To investigate the performance of suction cups, experiments have been conducted to clarify the effect of the pre-stretch ratio and chamber angle on suction forces. It could be seen that both factors have a complicated influence on suction forces, and the best performance obtained was a reasonable combination of the pre-stretch ratio and chamber angle. Here, we achieved a maximum suction force of 175 mN with λp = 1.2, α = 23° under a DC voltage of 3500 V. To enhance the capacity and adaptation of the suction cup, flat objects of various types of materials were introduced as targets. Experimental results displayed that for tested materials, including a dry/wet acrylic plate, CD, ceramic wafer, and aluminum plate, the suction cup showed outstanding performance of absorbing and lifting the target without any damage or scratch to them. Our research may serve as a guide to the optimal design and provide insights into the performance of the HCDEAs-actuated suction cup.

Humidity Effect on Dynamic Electromechanical Properties of Polyacrylic Dielectric Elastomer: An Experimental Study.

In this research, by utilizing the Very-High-Bond (VHB) 4905 elastomer, we carry out an experimental examination on the humidity effect on dynamic electromechanical performances of dielectric elastomers, including the dynamic response and viscoelastic creeping. Firstly, we experimentally analyze effects of the pre-stretch, peak voltage, waveform and frequency of the dynamic response of VHB 4905 elastomer under several ambient humidities. In general, the amplitude of dynamic deformation gradually adds up with the increasing humidity. Besides, it is found that the amplitude affected by different parameters shows diverse sensitivity to humidity. Subsequently, effect of humidity on the viscoelastic creeping of VHB 4905 is explored. The results demonstrate that, subject to different ambient humidities, the viscoelastic creeping under Alternating Current (AC) voltage is similar to that under Direct Current (DC) voltage. Furthermore, the equilibrium position of dynamic viscoelastic creep enlarges gradually with the humidity, regardless of voltage waveforms. For the dielectric elastomer with a pre-stretch ratio of 3, when the humidity increases from 20% to 80%, the increase of average equilibrium position of dynamic viscoelastic creep is larger than 1599%.