Autophagy regulation and protein kinase activity of PIK3C3 controls sertoli cell polarity through its negative regulation on SCIN (scinderin).

Sertoli cells are highly polarized testicular cells that provide a nurturing environment for germ cell development and maturation during spermatogenesis. The class III phosphatidylinositol 3-kinase (PtdIns3K) plays core roles in macroautophagy in various cell types; however, its role in Sertoli cells remains unclear. Here, we generated a mouse line in which the gene encoding the catalytic subunit, Pik3c3, was specifically deleted in Sertoli cells (cKO) and found that after one round of normal spermatogenesis, the cKO mice quickly became infertile and showed disruption of Sertoli cell polarity and impaired spermiogenesis. Subsequent proteomics and phosphoproteomics analyses enriched the F-actin cytoskeleton network involved in the disorganized Sertoli-cell structure in cKO testis which we identified a significant increase of the F-actin negative regulator SCIN (scinderin) and the reduced phosphorylation of HDAC6, an α-tubulin deacetylase. Our results further demonstrated that the accumulation of SCIN in cKO Sertoli cells caused the disorder and disassembly of the F-actin cytoskeleton, which was related to the failure of SCIN degradation through the autophagy-lysosome pathway. Additionally, we found that the phosphorylation of HDAC6 at site S59 by PIK3C3 was essential for its degradation through the ubiquitin-proteasome pathway. As a result, the HDAC6 that accumulated in cKO Sertoli cells deacetylated SCIN at site K189 and led to a disorganized F-actin cytoskeleton. Taken together, our findings elucidate a new mechanism for PIK3C3 in maintaining the polarity of Sertoli cells, in which both its autophagy regulation or protein kinase activities are required for the stabilization of the actin cytoskeleton.Abbreviations: ACTB: actin, beta; AR: androgen receptor; ATG14: autophagy related 14; BafA1: bafilomycin A1; BECN1: beclin 1, autophagy related; BTB: blood-testis barrier; CASP3: caspase 3; CDC42: cell division cycle 42; CDH2: cadherin 2; CHX: cycloheximide; CTNNA1: catenin (cadherin associated protein), alpha 1; CYP11A1: cytochrome P450, family 11, subfamily A, polypeptide 1; EBSS: Earle's balanced salt solution; ES: ectoplasmic specialization; FITC: fluorescein isothiocyanate; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; GCNA: germ cell nuclear acidic protein; GJA1: gap junction protein, alpha 1; H2AX: H2A.X variant histone; HDAC6: histone deacetylase 6; KIT: KIT proto-oncogene, receptor tyrosine kinase; LAMP1: lysosomal associated membrane protein 1; MAP3K5: mitogen-activated protein kinase kinase kinase 5; MAP1LC3B: microtubule associated protein 1 light chain 3 beta; OCLN: occludin; PIK3C3: phosphatidylinositol 3-kinase catalytic subunit type 3; PIK3R4: phosphoinositide-3-kinase regulatory subunit 4; PNA: arachis hypogaea lectin; RAC1: Rac family small GTPase 1; SCIN: scinderin; SQSTM1/p62: sequestosome 1; SSC: spermatogonia stem cell; STK11: serine/threonine kinase 11; TJP1: tight junction protein 1; TubA: tubastatin A; TUBB3: tubulin beta 3 class III; TUNEL: TdT-mediated dUTP nick-end labeling; UB: ubiquitin; UVRAG: UV radiation resistance associated gene; VIM: vimentin; WT1: WT1 transcription factor; ZBTB16: zinc finger and BTB domain containing 16.

Assessing Intrahospital Care Transition Structures Before and During the COVID-19 Pandemic: Network Analysis Study.

Transitions of care (TOC) is essential for patients with complex medical needs to maintain the continuity of care. The COVID-19 pandemic may result in unexpected pressure on healthcare organizations' routine work and may burden the TOC system. The objective of this study is to assess TOC structures in pre- and intra-COVID-19 and quantify changes in the structures through the lens of network analysis. We investigated a trauma registry repository consisting of care transitions of 5,674 (2,699 and 2,975 in pre- and intra-COVID-19) inpatients admitted to Vanderbilt University Medical Center (VUMC) between January 2019 and May 2021. Network metrics, including assortativity, homophily, and small-world-ness were leveraged to measure TOC structures and their changes. Our results showed both pre- and intra-COVID-19 TOC structures were disassortative, homophily, and small-world- ness, and the COVID-19 pandemic had limited influences on the three characteristics of the TOC structures. The disassortative TOC structure indicates patients can be efficiently transferred between triage centers (highly connected units) and receivers (lowly connected units); the homophily structure demonstrates two connected care units serve similar patients, and the small-world-ness reveals a patient can be transferred to highly collaborative care units with a short length of transfer path.

Protection of γ-Amino Butyric Acid on Radiation Induced Intestinal Injury in Mice.

SCOPE: Gamma-aminobutyric acid (GABA) represents positive effects in stress model, but the exact antioxidant remains unclear. This study aims to determine what GABA do and how GABA interfere on oxidative stress in the small intestine of radiation-induced intestinal injury (RIII) mice. METHODS AND RESULTS: C57BL/6J mice are gavaged. (1) Investigating the effects of GABA (50 100 mg kg-1  BW d-1 ) on basic information of healthy mice, and the survival time of RIII mice. (2) Evaluating the effect between GABA and theanine (100 mg kg-1  BW d-1 ) to RIII mice on the small intestine, by observing jejunum pathology, oxidative stress in small intestine and its mitochondria, and apoptosis protein expression. GABA reduces the weight loss and prolongs the median survival time of RIII mice. GABA and theanine reduce liver hyperemia, protect the villus crypt of jejunum, increase the antioxidant of duodenum and its mitochondria, to maintain the normal function and morphology. Besides, GABA increases B-cell lymphoma-2 (Bcl-2) expression and inhibits Caspase-3 activation, thereby inhibiting mitochondria-induced apoptosis. CONCLUSION: GABA reduces the oxidative stress of small intestine in RIII mice, and maintains the normal morphology and function of mitochondria, which mechanism is that high Bcl-2 expression inhibits the autophagy of mitochondrial pathway, thus reducing intestinal barrier damage.

Identifying Spatial Matching between the Supply and Demand of Medical Resource and Accessing Carrying Capacity: A Case Study of Shenzhen, China.

Previous Studies, such as the evaluation of the supply of and demand for regional medical resources and carrying capacity assessments, require further development. This paper aims to evaluate the carrying capacity and spatial distribution of medical resources in Shenzhen from the perspective of supply and demand, and to conduct a time-series variation of the coupling coordination degree from 1986 to 2019. The two-step floating catchment area method was employed to quantify the carrying capacity and coupling coordination degree method and spatial autocorrelation analysis were applied to analyze spatial distribution between supply and demand. The results were as follows. (1) The carrying capacity index in more than 50% of the districts was classified as low-grade. The percentage of regions with good grades was 8.27%. The regions with a high carrying capacity were distributed in the central and southeastern areas. (2) The coupling coordination continued to rise, increasing from 0.03397 in 1986 to 0.33627 in 2019. (3) The level of supply and demand for medical resources in Shenzhen increased from 1986 to 2019, and the highest degree of compatibility between the supply and the population size was largely concentrated in the western and eastern regions. This research can provide a theoretical reference for Shenzhen to rationally plan medical resources and improve the carrying capacity of medical resources.

Polyethylene Hydrogenolysis at Mild Conditions over Ruthenium on Tungstated Zirconia.

Plastics waste has become a major environmental threat, with polyethylene being one of the most produced and hardest to recycle plastics. Hydrogenolysis is potentially the most viable catalytic technology for recycling. Ruthenium (Ru) is one of the most active hydrogenolysis catalysts but yields too much methane. Here we introduce ruthenium supported on tungstated zirconia (Ru-WZr) for hydrogenolysis of low-density polyethylene (LDPE). We show that the Ru-WZr catalysts suppress methane formation and produce a product distribution in the diesel and wax/lubricant base-oil range unattainable by Ru-Zr and other Ru-supported catalysts. Importantly, the enhanced performance is showcased for real-world, single-use LDPE consumables. Reactivity studies combined with characterization and density functional theory calculations reveal that highly dispersed (WO x )n clusters store H as surface hydroxyls by spillover. We correlate this hydrogen storage mechanism with hydrogenation and desorption of long alkyl intermediates that would otherwise undergo further C-C scission to produce methane.

The complete chloroplast genome of Agrostis capillaris L.

Agrostis capillaris is a cool-season turf grass species that is found worldwide in temperate countries, and a good Pb phytostabilizer. In this study, the entire chloroplast genome sequence of A. capillaris was determined by Illumina sequencing. The complete chloroplast genome was circular and composed of 136,396 bp nucleotides with a GC content of about 38.5%. There were a large single-copy region (LSC, 81,659bp), a small single-copy region (SSC, 12,593bp), and a pair of reverse repeat regions (IRs, 42,144bp) in the chloroplast genome. In total, the A. capillaris chloroplast genome contained 144 genes, including 96 protein-coding genes, 40 tRNA genes, and 8 rRNA genes. Phylogenetic analysis revealed that A. capillaris was closely related to A. gigantean. The sequence data of A. capillaris chloroplast genome could provide useful genetic information for the studies on phylogenetic and evolutionary of Agrostidinae.

The complete chloroplast genome of Securigera varia (L.) Lassen.

Securigera varia is an important leguminous forage grass species that is mainly distributed in arid and semi-arid land with water scarcity, and has outstanding drought resistance. In this study, Illumina sequencing was used to obtain the complete sequence of the S. varia chloroplast genome. The complete genome was 154,257 bp in length with 35.9% GC content. It was a circular genome containing a large single-copy region (LSC, 84,762 bp), a small single-copy region (SSC, 18,059 bp), and a pair of inverted repeat regions (IRs, 51,436 bp). A total of 128 genes were encoded, including 83 protein-coding genes, 37 tRNAs, and 8 rRNAs. Phylogenetic analysis revealed that S. varia was closely related to Robinia pseudoacacia. The sequence data of S. varia chloroplast genome could provide useful genetic information for the studies on phylogenetic and evolutionary of Leguminosae.

Exogenous melatonin improves growth in hulless barley seedlings under cold stress by influencing the expression rhythms of circadian clock genes.

BACKGROUND: Melatonin is a hormone substance that exists in various living organisms. Since it was discovered in the pineal gland of cattle in 1956, the function of melatonin in animals has been roughly clarified. Nevertheless, in plants, the research on melatonin is still insufficient. Hulless barley (Hordeum vulgare L. var. nudum hook. f.) is a crop that originates from cultivated barley in the east, usually grown on the Qinghai-Tibet Plateau, becoming the most important food crop in this area. Although the genome and transcriptome research of highland barley has gradually increased recently years, there are still many problems about how hulless barley adapts to the cold climate of the Qinghai-Tibet Plateau. METHODS: In this study, we set three temperature conditions 25°C, 15°C, 5°C hulless barley seedlings, and at the same time soaked the hulless barley seeds with a 1 µM melatonin solution for 12 hours before the hulless barley seeds germinated. Afterwards, the growth and physiological indicators of hulless barley seedlings under different treatment conditions were determined. Meanwhile, the qRT-PCR method was used to determine the transcription level of the hulless barley circadian clock genes under different treatment conditions under continuous light conditions. RESULTS: The results showed the possible mechanism by which melatonin pretreatment can promote the growth of hulless barley under cold stress conditions by studying the effect of melatonin on the rhythm of the circadian clock system and some physiological indicators. The results revealed that the application of 1 µM melatonin could alleviate the growth inhibition of hulless barley seedlings caused by cold stress. In addition, exogenous melatonin could also restore the circadian rhythmic oscillation of circadian clock genes, such as HvCCA1 and HvTOC1, whose circadian rhythmic phenotypes were lost due to environmental cold stress. Additionally, the results confirmed that exogenous melatonin even reduced the accumulation of key physiological indicators under cold stress, including malondialdehyde and soluble sugars. DISCUSSION: Overall, these findings revealed an important mechanism that exogenous melatonin alleviated the inhibition of plant vegetative growths either by restoring the disrupted circadian rhythmic expression oscillations of clock genes, or by regulating the accumulation profiles of pivotal physiological indicators under cold stress.

[Spatiotemporal variability characteristics and driving forces of land use in the Pan-Pearl River Basin, China]. / æ³›ç æ±ŸæµåŸŸåœŸåœ°åˆ©ç”¨æ—¶ç©ºå˜åŒ–ç‰¹å¾åŠé©±åŠ¨å› å­.

The Pan-Pearl River Basin is a bridgehead for China's reform and opening-up and the construction of the Belt and Road at Sea, with vital strategic significance in Chinese overall development. Land use data and climate and socio-economic indicators were integrated to probe the spatiotemporal change and its driving forces of land use in the Pan-Pearl River basin with ArcGIS spatial analysis tool and SPSS factor analysis tool. Results showed that land use in the Pan-Pearl River Basin significantly changed between 1990 and 2015, with decreases of the area of paddy field and woodland and rapid increases of urban land and other construction land. Outflow of grassland occurred in the northwestern part of the basin. Reduction of cultivated field was mainly concentrated in the central part of the basin and coastal areas. Increases in urban and rural land, industrial and mining land, and residential land were centrally distributed in the Guangdong-Hong Kong-Marco Greater Bay Area. The prominent change areas were Guangdong-Hong Kong-Marco Greater Bay Area : central and southeast of Guangxi Province : northern Hainan Province. Land use changes during 1990-2000 were most obvious in the basin. The main driving factor of spatiotemporal variation of land use was the rapid development of social economy and industry and the improvement of residents' consumption level.

Temporal microstructure of dyadic social behavior during relationship formation in mice.

Socially competent animals must learn to modify their behavior in response to their social partner in a contextually appropriate manner. Dominant-subordinate relationships are a particularly salient social context for mice. Here we observe and analyze the microstructure of social and non-social behaviors as 21 pairs of outbred CD-1 male mice (Mus Musculus) establish dominant-subordinate relationships during daily 20-minute interactions for five consecutive days in a neutral environment. Firstly, using a Kleinberg burst detection algorithm, we demonstrate aggressive and subordinate interactions occur in bursting patterns followed by quiescent periods rather than being uniformly distributed across social interactions. Secondly, we identify three phases of dominant-subordinate relationship development (pre-, middle-, and post-resolution) by utilizing two statistical methods to identify stability in aggressive and subordinate behavior across these bursts. Thirdly, using First Order Markov Chains we find that dominant and subordinate mice show distinct behavioral transitions, especially between tail rattling and other aggressive/subordinate behaviors. Further, dominant animals engaged in more digging and allogrooming behavior and were more likely to transition from sniffing their partner's body to head, whereas subordinates were more likely to transition from head sniffing to side-by-side contact. Lastly, we utilized a novel method (Forward Spike Time Tiling Coefficient) to assess how individuals respond to the behaviors of their partner. We found that subordinates decrease their tail rattling and aggressive behavior in response to aggressive but not subordinate behavior exhibited by dominants and that tail rattling in particular may function to deescalate aggressive behavior in pairs. Our findings demonstrate that CD-1 male mice rapidly establish dominance relationships and modify their social and non-social behaviors according to their current social status. The methods that we detail also provide useful tools for other researchers wishing to evaluate the temporal dynamics of rodent social behavior.