Roundup® induces premature senescence of mouse granulosa cells via mitochondrial ROS-triggered NLRP3 inflammasome activation.

Roundup, a glyphosate-based herbicide widely used in agriculture, has raised concerns regarding its potential impact on human health due to the detection of its residues in human urine and serum. Granulosa cells are essential for oocyte growth and follicle development. Previous research has shown that Roundup could affect steroid synthesis, increases oxidative stress, and induces apoptosis in granulosa cells. However, little is known about the effects of Roundup on NLRP3 (nucleotide binding oligomerization domain-like receptor family pyrin-containing domain protein 3) inflammasome activation and cellular senescence in granulosa cells. Here, we provided evidence that exposure to Roundup induced premature senescence in mouse granulosa cells through the activation of NLRP3 inflammasome triggered by mitochondrial ROS. Our findings demonstrated that Roundup significantly reduced the viability of granulosa cells under in vitro culture conditions. It also disrupted mitochondrial function and induced oxidative stress in these cells. Subsequent investigations showed that NLRP3 inflammasome was activated in treated granulosa cells, as evidenced by the upregulation of inflammasome-related genes and the processing of inflammatory cytokines IL-1ß and IL-1α into their mature forms. Consequently, premature cellular senescence occurred in response to the challenge posed by Roundup. Notably, direct inhibition of NLRP3 inflammasome with MCC950 does not alleviate mitochondrial damage and oxidative stress. However, supplementation of resveratrol, which has been known to attenuate mitochondrial damage and oxidative stress, effectively mitigated the inflammatory response and the expression of senescence-related markers, and prevented the senescence in granulosa cells. These results suggested that mitochondrial function and oxidative homeostasis might play pivotal roles as upstream regulators of NLRP3 inflammasome. In summary, our findings indicated that the premature senescence of granulosa cells caused by mitochondrial ROS-triggered NLRP3 inflammasome activation might contribute to the ovarian toxicity of Roundup, in addition to its known effects on steroidogenesis and apoptosis. Supplementary Information: The online version contains supplementary material available at 10.1007/s43188-024-00229-0.

PATJ and MPDZ are required for trophectoderm lineage specification in early mouse embryos.

In brief: During the morula to blastocyst transformation, polarity establishment in outer cells is a prerequisite for trophectoderm lineage specification. This study reveals the roles of polarity proteins PATJ and MPDZ in trophectoderm lineage fate decision. Abstract: In mouse preimplantation embryos, cell polarity plays a crucial role in the first lineage specification. PATJ and its homolog MPDZ are the main members of CRB-PALS1-PATJ (CRUMBS-Protein associated with Lin7 1-Pals-associated tight junction protein) apical polarity complex. They act as adaptor proteins connecting CRB-PALS1 and tight junction proteins, making them essential for cell polarization and stabilization of apical junctions. However, their roles in regulating trophectoderm differentiation and blastocyst development remain unclear. In this study, PATJ and/or MPDZ were downregulated by the microinjection of specific RNA interference constructs into zygotes. Downregulation of PATJ alone did not severely affect early embryonic development and trophectoderm lineage differentiation although it slowed down the blastocyst formation. Depletion of PATJ and MPDZ did not affect compaction and morula development but impaired blastocyst formation. Furthermore, the expression of trophectoderm-specific transcription factors and trophoblast differentiation was compromised in the absence of PATJ/MPDZ. These abnormalities might result from the breakdown of apical domain in the outer cells of the embryo. The loss of PATJ/MPDZ caused the breakdown of CRB and PAR polarity complexes as well as deficiencies in tight junctions and actin filaments. These defects led to ectopic activation of Hippo signaling in the outer cells of developing embryos, ultimately suppressing Cdx2 expression and trophectoderm differentiation. Altogether, PATJ and MPDZ are essential for trophectoderm lineage differentiation and normal blastocyst morphogenesis via the regulation of the establishment of apical domain, formation of tight junctions, phosphorylation and localization of YAP, and expression of trophectoderm-specific transcription factors.

Noninvasive autologous mitochondria transport improves the quality and developmental potential of oocytes from aged mice.

OBJECTIVE: To establish an optimized autologous mitochondria transport technique for oocyte-aging rescue, which minimizes both the patient's pains and the damage to oocytes. DESIGN: Experimental laboratory study. SETTING: Laboratory. ANIMAL(S): Institute of Cancer Research mice. INTERVENTION(S): The murine umbilical cord mesenchymal stem cells were isolated from the female pup and cryopreserved. After the female aged, its germinal vesicle (GV) oocytes were collected and treated to weaken the zona pellucida. Its autologous umbilical cord mesenchymal stem cells were induced into granulosa cells (iGCs). The zona-weakened GV oocytes were aggregated with iGCs into iGC-oocyte complexes. Then, these complexes were cultured in growth-differentiation factor 9-containing media for 3 days. Next, they were subjected to in vitro maturation and fertilization. Presumptive zygotes were cultured for 24 hours, and the cleaved 2-cell embryos were selected for embryo transfer. MAIN OUTCOME MEASURE(S): The oocyte quality was determined by examining mitochondrial ultrastructure using transmission electron microscopy, the adenosine triphosphate content using a luminometer, and intracellular reactive oxygen species levels by confocal microscopy. The spindle organization in mature oocytes was examined by confocal microscopy. The developmental potential of oocytes was evaluated by monitoring the in vitro embryo development and the birth rate after embryo transfer. RESULT(S): Mitochondria migrated from iGCs into the GV oocyte via transzonal filopodia. The maturation rate, quality, and developmental potential of these oocytes were substantially increased. Furthermore, the birth rate after embryo transfer has been improved. CONCLUSION(S): This approach used noninvasive procedures to collect mitochondria donor cells and optimized mitochondria transfer manipulations; thus, it may have potential in ameliorating oocyte-aging-related subfertility.

Elimination of abnormal phase fluctuation in digital holography.

In digital holography, the phase is most important, and the quality of the reconstructed phase determines the final reconstructed image effect. However, noise is inevitably introduced in the process of recording the hologram. For regions without object light, the phase has a random distribution, which affects the final phase quality. This kind of noise is called abnormal phase fluctuations in this paper. The correlation between amplitude and phase in digital holography is used to judge whether there is useful phase information. Through structural similarity and the light-dark relationship, a credible probability mask is introduced to extract the phase that needs to be preserved. The simulation and experimental results show that abnormal phase fluctuations are successfully removed, and the useful phase information is retained.

Speckle reduction in digital holography with low-dimensional reconstruction.

Speckle reduction is a crucial technique since the presence of speckle disturbs the quality of the reconstruction in digital holography. In this paper, we present an easy, fast, and efficient single-shot method to reduce speckle noise in digital holography. The method reconstructs subholograms from a single hologram. Then, sub-reconstruction images are randomly shuffled and divided into several groups and low-dimensional noise-reduced images can be achieved by averaging sub-reconstruction image groups by groups. Next, these low-dimensional noise-reduced images are combined to obtain a noise-reduced image. Finally, the noise-reduced image is processed by a mean filter to obtain a final image, which has substantially less speckle noise while preserving the dimensions of the original image. The experimental results demonstrate the effectiveness of the proposed method and indicate its potential in real-time digital holography.

Single-shot speckle reduction by elimination of redundant speckle patterns in digital holography.

Speckle reduction is a crucial technique, since the presence of speckle disturbs the quality of the reconstruction in digital holography. In this paper, we present a redundant speckle elimination method to suppress the speckle noise. For the same position in each of the reconstructed sub-images, we consider pixels with the same gray value as information with the same speckle distribution. Therefore, a speckle-suppressed gray value can be obtained by extracting pixels with different gray values and then averaging. Through theoretical analysis and experiments, we demonstrate that speckle contrast can be decreased significantly by using the proposed method. Moreover, we show that the despeckle strength of the proposed method highly depends on the number of binary masks. These results indicate the potential of the proposed method for various applications.

Erratum: Bao, L.; Cai, W.; Zhang, X.; Liu, J.; Chen, H.; Wei, Y.; Jia, X.; Bai, Z. Distinct Microbial Community of Phyllosphere Associated with Five Tropical Plants on Yongxing Island, South China Sea. Microorganisms 2019, 7, 525.

The authors wish to make the following erratum in this paper [...].

Microbial community overlap between the phyllosphere and rhizosphere of three plants from Yongxing Island, South China Sea.

Phyllosphere and rhizosphere are unique and wide-ranging habitats that harbor various microbial communities, which influence plant growth and health, and the productivity of the ecosystems. In this study, we characterized the shared microbiome of the phyllosphere and rhizosphere among three plants (Ipomoea pes-caprae, Wedelia chinensis, and Cocos nucifera), to obtain an insight into the relationships between bacteria (including diazotrophic bacteria) and fungi, present on these host plants. Quantitative PCR showed that the abundances of the microbiome in the soil samples were significantly higher than those in the phyllosphere samples, though there was an extremely low abundance of fungi in bulk soil. High-throughput sequencing showed that the alpha-diversity of bacteria and fungi was higher in the rhizosphere than the phyllosphere samples associated with the same plant, while there was no obvious shift in the alpha-diversity of diazotrophic communities between all the tested phyllosphere and soil samples. Results of the microbial composition showed that sample-specific bacteria and fungi were found among the phyllosphere and rhizosphere of the different host plants. About 10%-27% of bacteria, including diazotrophs, and fungi overlapped between the phyllosphere and the rhizosphere of these host plants. No significant difference in microbial community structure was found among the tested rhizosphere samples, and soil properties had a higher influence on the soil microbial community structures than the host plant species.

Low-dose Roundup induces developmental toxicity in bovine preimplantation embryos in vitro.

Roundup is a widely used glyphosate-based herbicide worldwide. Roundup residues can be detected in the organs and urine of animals. However, its toxicity on mammalian preimplantation embryos has not been well investigated. Here, we show Roundup impairs the development and quality of bovine preimplantation embryos in a dose-dependent manner. Exposure to the agricultural recommended doses of Roundup caused in vitro developmental arrest and quick death of bovine embryos. Furthermore, even a very low concentration (0.9 ppm) of Roundup was harmful to bovine preimplantation development. In addition, Roundup increases intracellular calcium levels and induces oxidative stress and apoptosis in bovine embryos. Even if the embryos developed to morphologically normal blastocysts when cultured with low concentrations of Roundup, abnormal intracellular calcium and oxidative stress could be detected inside the embryos and led to an increased incidence of apoptosis in the blastocysts. These data suggest Roundup residues from the agricultural application are potentially dangerous to mammalian preimplantation embryos.

Seasonal variation of epiphytic bacteria in the phyllosphere of Gingko biloba, Pinus bungeana and Sabina chinensis.

Phyllosphere harbors diverse microorganisms, which influence plant growth and health. In order to understand the extent to which environmental factors affect epiphytic microbial communities, we characterized microbial communities on leaves of three separate tree species present on the college campus, and also present within a forest park over two seasons. Quantitative PCR analysis showed the quantity of 16S rRNA genes was lower in May compared with October, while the abundances of functional genes (nifH and bacterial amoA genes) were extremely high in May. High-throughput sequencing revealed a large variation in the diversity and composition of bacterial and diazotrophic communities over the two seasons, and showed the abundance of functional genera, such as Nocardioides, Bacillus and Zoogloea were significantly elevated in May. In addition, xenobiotic biodegradation pathways of bacterial communities were clearly elevated in May. Network analysis showed the correlations between phyllospheric bacteria in May were more complex than that in October and showed greater negative correlations. These results were consistent in all tree species in this study. This study showed that phyllospheric bacteria varied greatly in different seasons, which implies that different growing seasons should be considered in the exploitation of the interactions between phyllospheric microorganisms and host plants.

Distinct Microbial Community of Phyllosphere Associated with Five Tropical Plants on Yongxing Island, South China Sea.

The surfaces of a leaf are unique and wide habitats for a microbial community. These microorganisms play a key role in plant growth and adaptation to adverse conditions, such as producing growth factors to promote plant growth and inhibiting pathogens to protect host plants. The composition of microbial communities very greatly amongst different plant species, yet there is little data on the composition of the microbiome of the host plants on the coral island in the South China Sea. In this study, we investigated the abundances and members of a major microbial community (fungi, bacteria, and diazotrophs) on the leaves of five dominant plant species (Ipomoea pes-caprae, Wedelia chinensis, Scaevola sericea, Cocos nucifera, and Sesuvium portulacastrum) on the island using real-time quantitative polymerase chain reaction (PCR) and high-throughput amplicon sequencing. Quantitative PCR results showed that fungi and bacteria were ubiquitous and variable among different host plants. Scaevola sericea showed the lowest absolute abundance and highest diversity of fungi and bacteria, while Cocos nucifera had the lowest abundance and the highest diversity of diazotrophs compare to the other four plants. There was a small proportion of shared microorganisms among the five different plants, while unique fungi, bacteria and diazotrophs were significantly enriched for different host plant species in this study (p < 0.05). Some of the most abundant organisms found in the communities of these different host plants are involved in important biogeochemical cycles that can benefit their host, including carbon and nitrogen cycles.

Critical Care Resources in Guangdong Province of China: Three Surveys from 2005 to 2015.

OBJECTIVES: Data about the critical care resources in China remain scarce. The purpose of this study was to investigate the variation and distribution of critical care resources in Guangdong province from 2005 to 2015. DESIGN: Data in regard to critical care resources were collected through questionnaires and visits every 5 years from 2005. SETTING: All hospitals in Guangdong province were screened and hospitals that provide critical care services were enrolled. INTERVENTION: None. MEASUREMENTS AND MAIN RESULTS: One hundred eleven, 158, and 284 hospitals that provide critical care services were enrolled in the three consecutive surveys respectively. The number of ICUs, ICU beds, intensivists, and nurses increased to 324, 3,956, 2,470, and 7,695, respectively, by 2015. Adjusted by population, the number of ICU beds per 100,000 (100,000) population increased by 147.7% from 2005 to 2015, and the number of intensivists and nurses per 100,000 population increased by 35.3% and 55.1% from 2011 to 2015. However, the numbers in the Pearl River Delta, a richer area, were higher than those in the non-Pearl River Delta area (ICU beds: 4.64 vs 2.58; intensivists: 2.90 vs 1.61; nurses: 9.30 vs 4.71 in 2015). In terms of staff training, only 17.85% of intensivists and 14.29% of nurses have completed a formal accredited critical care training program by 2015. CONCLUSIONS: Our study was the first one to investigate the trend and distribution of critical care resources in China. The quantity of ICU beds and staff has been increasing rapidly, but professional training for staff was inadequate. The distribution of critical care resources was unbalanced. Our study can be beneficial for healthcare policymaking and the allocation of critical care resources in Guangdong province and other provinces in China.