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Using small molecules to induce readthrough of premature termination codons is a promising therapeutic approach to treating genetic diseases and cancers caused by nonsense mutations, as evidenced by the widespread use of ataluren to treat nonsense mutation Duchene muscular dystrophy. Herein we describe a series of novel guanidino quinazoline and pyrimidine scaffolds that induce readthrough in both HDQ-P1 mammary carcinoma cells and mdx myotubes. Linkage of basic, tertiary amines with aliphatic, hydrophobic substituents to the terminal guanidine nitrogen of these scaffolds led to significant potency increases. Further potency gains were achieved by flanking the pyrimidine ring with hydrophobic substituents, inducing readthrough at concentrations as low as 120 nM and demonstrating the potential of these compounds to be used either in combination with ataluren or as stand-alone therapeutics.

Ischemia-reperfusion injury (IRI) is the key reason of injury after testicular torsion and may eventually lead to male infertility. Oleuropein, a natural antioxidant isolated from Olea europaea, has shown beneficial effects in different models of ischemia. We evaluated the effects of oleuropein on testicular IRI and explored the underlying protective mechanisms.A mouse testicular torsion/detorsion (T/D) model and an oxygen-glucose deprivation/reperfusion (OGD/R) germ cell model were established and treated with oleuropein. HE staining was used to evaluate testicular pathological changes. Apoptosis and apoptosis-associated protein levels in testis tissues were assessed by TUNEL staining, immunohistochemical staining and western blot. Apoptosis levels and apoptosis-associated protein levels in GC-1 were evaluated by flow cytometry, immunofluorescence and western blot. Oxidative stress levels were assessed by malondialdehyde (MDA) and superoxide dismutase (SOD) kits. Cell viability and inflammatory protein levels were evaluated by CCK-8 assay coupled with qRT-PCR.Relative to the control group, SOD activity was markedly suppressed, while MDA, Bax, Caspase-3, TNF-α as well as IL-1β levels were significantly increased in the T/D model and OGD/R model. However, all of the aforementioned alterations were relieved by oleuropein treatment.Our findings indicate that oleuropein may be a promising treatment option to attenuate testicular IRI via its anti-oxidant, anti-inflammatory as well as anti-apoptotic properties.

Objective(s): The current study was aimed to investigate the effect of morpholin-4-ium 4 methoxyphenyl (morpholino) phosphinodithioate (GYY4137) on ipsilateral epididymis injury in a rat model of experimental varicocele (VC). Materials and Methods: Sixty Wistar rats were randomly assigned to sham, sham plus GYY4137, VC and VC plus GYY4137 groups. Sperm quality parameters, including sperm count, motility and viability were evaluated after 4 weeks. Histological changes were measured by hematoxylin and eosin staining between the groups. The oxidative stress levels were estimated by determining epididymal superoxide dismutase (SOD) and malondialdehyde (MDA). The apoptosis status and the expression of phosphatidylinositol 3′-OH kinase (PI3K)/Akt were analyzed by immunohistochemical analysis, western blot and RT-qPCR. Results: VC resulted in the decrease of sperm parameters, significant histological damage and higher levels of oxidative stress and apoptosis. Compared to the VC group, GYY4137 markedly ameliorated these observed changes. In addition, treatment with GYY4137 obviously reduced the levels of caspase-3 and Bax and increased the levels of the phosphorylation of PI3K p85 and Akt. Conclusion: Our data demonstrated that GYY4137 may alleviate the sperm damage and epididymis injury in experimentally VC-induced rats by activation of the PI3K/Akt pathway.

Background Potassium (K), an important nutrient element, can improve the stress resistance/tolerance of crops. The application of K in resisting plant parasitic nematodes shows that the K treatment can effectively reduce the occurrence of nematode diseases and increase crop yield. However, data on K2SO4 induced rice resistance to Meloidogyne graminicola are still lacking. To evaluate rice resistance against M. graminicola induced by K2SO4 and to further clarify its mechanism is essential for the rational use of K fertilizer to ensure the safety of rice production.Results In this work, K2SO4 treatment effectively reduced the numbers of both galls and nematodes in rice roots, and delayed the development of nematodes to the adult stage. Rather than by affecting the attractiveness of roots to nematodes and the morphological phenotype of giant cells at feeding sites, such effect was achieved by rapidly stimulating hydrogen peroxide (H2O2) accumulation, increasing callose deposition. Meanwhile, such induced resistance required the active participation of the potassium channel OsAKT1 and the potassium transporter OsHAK5. The numbers of both galls and nematodes were higher in both gene deficient plants than that in the wild-type plants, and the K2SO4-induced resistance showed weaker in the defective plants than in the wild-type plants.Conclusions K2SO4 treatment effectively induces rice resistance to root-knot nematode M. graminicola. The mechanism of inducing resistance is to prime the basic defense of rice, up-regulating the expression of resistance-related genes and with the involvement of K+ channel and transporter. These laid a foundation for further study on the mechanism of rice to defense against root-knot nematodes and the effective use of potassium fertilizer to improve rice resistance against nematodes in the field.

Ausgehend von der Situation der deutschen Sprache sowie des schulischen Unterrichts Deutsch als Fremdsprache in Malaysia wird in dieser Arbeit untersucht, ob das Curriculum des ersten Bachelorprogramms „German with Education“ (Bachelor of Language and Linguistics – German Language with Education), welches von 2009 bis 2017 an der Universität Malaya angeboten wurde, malaysische DaF-Lehrkräfte erfolgreich ausbilden und anschließend an die Sekundarschulen Malaysias vermitteln konnte. Die Arbeit umfasst insgesamt neun Kapitel. Das erste Kapitel stellt die spezifischen Bildungsbedingungen Malaysias dar. Das zweite Kapitel konzentriert sich auf den Einfluss einer multilingualen und multikulturellen Lernumgebung auf den Fremdspracherwerb. Im Mittelpunkt des dritten Kapitels steht die Darstellung des ersten Bachelorprogramms „German with Education“ zur Lehrkräftebildung im Fach Deutsch als Fremdsprache, das an der Universität Malaya entwickelt wurde. Im vierten Kapitel wird das Forschungsdesign zur Evaluation des Bachelorprogramms „German with Education“ erläutert. Im fünften Kapitel werden die Ergebnisse der Auswertung der Perspektive I bzw. die quantitativen und qualitativen Untersuchungen mitsamt den Ergebnissen der Befragung und Interviews der Absolventen/-innen und Studierenden dargestellt. Im sechsten Kapitel werden als Auswertung der Perspektive II die Ansichten und Erfahrungen externer und interner ministerieller Experten/-innen dargestellt. Im siebten Kapitel geht es in Auswertung der Perspektive III um die Ansichten und Lehrerfahrungen der DaF-Dozent/-innen und Lehrkräfte des Bachelorprogramms. Im achten Kapitel erfolgt die thematische Zusammenführung der Evaluationsergebnisse zum Bachelorprogramm. Im neunten Kapitel werden aus den Evaluationsergebnissen Empfehlungen zum Bachelorprogramm „German with Education“ generiert. Die Forschungsergebnisse der Arbeit werden abschließend dem Erziehungsministerium zur Verfügung gestellt, um die Neugestaltung des DaF-Fremdsprachencurriculums auf wissenschaftlicher Grundlage zu unterstützen.

To investigate the expression of long non-coding RNA (lncRNA) H19 in mouse GC-1 cells in vitro and its effects on the proliferation and apoptosis of GC-1 cells.We established an in vitro hypoxia-reoxygenation model in GC-1 cells and detected the expression of lncRNA H19 in the GC-1 cells at different time points of reoxygenation injury by qRT-PCR. We determined the effects of silencing lncRNA H19 on the proliferation and apoptosis of the GC-1 cells by MTT and flow cytometry, the expressions of apoptosis-related proteins Bax and caspase-3 in the GC-1 cells by Western blot, and the expressions of microRNA-203a and PTEN by qRT-PCR and Western blot, respectively.With the prolonging of the time of reoxygenation injury, the expression of lncRNA H19 was increased significantly in the GC-1 cells and peaked at 3-hour hypoxia and 12-hour reoxygenation, but that of microRNA-203a markedly decreased. Silencing lncRNA H19 enhanced the proliferation and inhibited the apoptosis of the GC-1 cells, and up-regulated the expression of microRNA-203a and down-regulated that of PTEN in the GC-1 cells.LncRNA H19 is highly expressed in GC-1 cells in vitro, which may influence the proliferation and apoptosis of GC-1 cells by regulating the microRNA-203a /PTEN signaling pathway.

The outbreak of coronavirus disease 2019 (COVID-19) caused by 2019 novel coronavirus has become a global public health challenge. In addition to the typical respiratory symptoms, COVID-19 can induce damage to testicular spermatogenesis. This study focuses on the possible causes and follow-up monitoring of testicular injury induced by COVID-19.

To study the effect of different levels of autophagy in the testis on the apoptosis of spermatogenic cells in the rat model of varicocele (VC).We randomly divided 54 SD male rats into six groups, blank control (n = 6), rapamycin control (n = 6), chloroquine control (n = 6), VC model control (n = 12), VC + rapamycin (n = 12), and VC + chloroquine (n = 12). We observed the histomorphological changes of the testis and epididymis by HE staining, obtained the scores on spermatogenesis in the testis and epididymis, calculated the apoptosis index (AI) of the testicular spermatogenic cells by TUNEL, and determined the expressions of LC3-Ⅱ, LC3-Ⅰ, p62, Bax and Bcl-2 proteins in the testis tissue by Western blot.There were no significant morphological changes in the testis and epididymis of the rats in the blank control, rapamycin control and chloroquine control groups, or significant differences in the scores on testicular and epididymal spermatogenesis and AI of the testicular spermatogenic cells (P0.05). The animals in the VC model control group exhibited significant pathological damage in the testicular and epididymal tissues, with remarkably decreased scores on spermatogenesis (P0.01) and increased AI (P0.01), which were markedly improved in the VC + rapamycin group and slightly aggravated in the VC + chloroquine group compared with the VC model controls. In comparison with the rats in the blank control group, those in the VC model control group showed significantly up-regulated expressions of the autophagy-related protein LC3 (including the LC3-Ⅱ/LC3-Ⅰ ratio) and the pro-apoptotic protein Bax in testicular tissue (P0.01) but down-regulated expression of the anti-apoptotic protein Bcl-2 (P0.01). The expressions of LC3 and Bcl-2 in the testis tissue were significantly higher in the VC + rapamycin (P0.01) but lower in the VC + chloroquine group (P0.01), while those of p62 and Bax remarkably lower in the VC + rapamycin (P0.01) but higher in the VC + chloroquine group than in the VC model controls (P0.01).Varicocele induces autophagy in the testis and apoptosis of spermatogenic cells in rats. Up-regulating autophagy can inhibit while blocking autophagy can promote the apoptosis of spermatogenic cells.

Guided bone regeneration in inflammatory microenvironments of osteoporotic patients with large alveolar bone defects remains a great challenge. Macrophages are necessary for alveolar bone regeneration via their polarization and paracrine actions. Our previous studies showed that Cu-bearing Ti6Al4V alloys are capable of regulating macrophage responses. When considering the complexity of oral microenvironments, the influences of Cu-bearing Ti6Al4V alloys on osteoporotic macrophages in infectious microenvironments are worthy of further investigations. In this study, we fabricated Ti6Al4V-Cu alloy by selective laser melting technology and used Porphyromonas gingivalis lipopolysaccharide (P.g-LPS) to imitate oral pathogenic bacterial infections. Then, we evaluated the impacts of Ti6Al4V-Cu on osteoporotic macrophages in infectious microenvironments. Our results indicated that Ti6Al4V-Cu not only inhibited the P.g-LPS-induced M1 polarization and pro-inflammatory cytokine production of osteoporotic macrophages but also shifted polarization towards the pro-regenerative M2 phenotype and remarkably promoted anti-inflammatory cytokine release. In addition, Ti6Al4V-Cu effectively promoted the activity of COMMD1 to potentially repress NF-κB-mediated transcription. It is concluded that the Cu-bearing Ti6Al4V alloy results in ameliorated osteoporotic macrophage responses to create a favourable microenvironment under infectious conditions, which holds promise to develop a GBR-barrier membrane for alveolar bone regeneration of osteoporosis patients.

The potential for endophytes to initiate changes in host secondary metabolism has been well documented. However, the mechanisms underlying endophyte-plant metabolic interactions are still poorly understood. Here, we analysed the effects of fungal endophytes on the metabolite profiles of grape cells from two cultivars: 'Cabernet Sauvignon' (CS) and 'Rose honey' (RH). Our results clearly showed that co-culture with endophytic fungi greatly modified the metabolic profiles in grape cells of both varieties. Treatments with endophytic fungal strains caused the numbers of detected metabolites to vary from 10 to 19 in CS cells and from 8 to 14 in RH cells. In addition, 5 metabolites were detected in all CS cell samples, while 4 metabolites were detected in all RH cell samples. Some endophytic fungal strains could even introduce novel metabolites into the co-cultured grape cells. The metabolic profiles of grape leaves shaped by endophytic fungi exhibited host selectivity and fungal strain specificity. In this assay, the fungal strains RH32 (Alternaria sp.) and MDR36 (Colletotrichum sp.) triggered an increased response of the detected metabolites, including the greatest increase in the metabolite contents in grape cells of both cultivars. No obvious effects in terms of metabolite numbers and contents in grape cells when co-cultured with fungal strains RH7 (Epicoccum sp.) and RH48 (Colletotrichum sp.) were observed. The results of this experiment suggest that endophytic fungi could be used to control the metabolic profiles of grapes and thus increase grape quality., VITIS - Journal of Grapevine Research, Vol. 59 No. 4 (2020): Vitis

The molecular mechanism by which plants defend against plant root-knot nematodes (RKNs) is largely unknown. The plant receptor kinase FERONIA and its peptide ligands, rapid alkalinization factors (RALFs), regulate plant immune responses and cell expansion, which are two important factors for successful RKN parasitism. In this study, we found that mutation of FERONIA in Arabidopsis thaliana resulted in plants showing low susceptibility to the RKN Meloidogyne incognita. To identify the underlying mechanisms associated with this phenomenon, we identified 18 novel RALF-likes from multiple species of RKNs and showed that two RALF-likes (i.e., MiRALF1 and MiRALF3) from M. incognita were expressed in the esophageal gland with high expression during the parasitic stages of nematode development. These nematode RALF-likes also possess the typical activities of plant RALFs and can directly bind to the extracellular domain of FERONIA to modulate specific steps of nematode parasitism-related immune responses and cell expansion. Genetically, both MiRALF1/3 and FERONIA are required for RKN parasitism in Arabidopsis and rice. Collectively, our study suggests that nematode-encoded RALFs facilitate parasitism via plant-encoded FERONIA and provides a novel paradigm for studying host–pathogen interactions.

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Massive oral and maxillofacial bone defect regeneration remains a major clinical challenge due to the absence of functionalized bone grafts with ideal mechanical and pro-regeneration properties. In the present study, Laponite (LAP), a synthetic nanosilicate, was incorporated into polycaprolactone (PCL) to develop a biomaterial for bone regeneration. We explored whether LAP-embedded PCL would accelerate bone regeneration by orchestrating osteoblasts to directly and indirectly induce bone regeneration processes. The results confirmed the presence of LAP in PCL, and LAP was distributed in the exfoliated structure without aggregates. Incorporation of LAP in PCL slightly improved the compressive properties. LAP-embedded PCL is biocompatible and exerts pronounced enhancements in cell viability, osteogenic differentiation and extracellular matrix formation of osteoblasts. Furthermore, osteoblasts cultured on LAP-embedded PCL facilitated angiogenesis of vessel endothelial cells and alleviated osteoclastogenesis of osteoclasts in a paracrine manner. The addition of LAP to the PCL endowed favorable bone formation in vivo. Based upon these results, LAP-embedded PCL shows great potential as an ideal bone graft that exerts both space-maintaining and vascularized bone regeneration synergistic effects and can be envisioned for oral and maxillofacial bone defect regeneration. This article is protected by copyright. All rights reserved.