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Polysaccharide is one of the principal bioactive components found in medicinal mushrooms and has been proven to enhance host immunity. However, the possible mechanism of immunomodulatory activity of Cordyceps militaris polysaccharide is not fully understood. Hot water extraction and alcohol precipitation, DEAE-Sephadex A-25 chromatography, and Sephadex G-100 chromatography were used to isolate polysaccharide from C. militaris. A high-molecular-weight polysaccharide isolated from C. militaris was designated as HCMP, which had an Mw of 6.18 × 105 Da and was composed of arabinose, galactose, glucose, mannose, and xylose in a mole ratio of 2.00:8.01:72.54:15.98:1.02. The polysaccharide content of HCMP was 91.2% ± 0.16. The test in vitro showed that HCMP activated mouse macrophage RAW 264.7 cells by enhancing phagocytosis and NO production, and by regulating mRNA expressions of inflammation-related molecules in RAW 264.7 cells. Western blotting revealed that HCMP induced the phosphorylation of mitogen-activated protein kinases (MAPKs). Moreover, using inhibitors of MAPKs decreased the mRNA levels of inflammation-related molecules induced by HCMP. These data evidenced that the immunomodulatory effect of HCMP on RAW 264.7 macrophages was mediated via the MAPK signaling pathway. These findings suggested that HCMP could be developed as a potent immunomodulatory agent for use in functional foods and dietary supplements. [ABSTRACT FROM AUTHOR]

Background People who inject drugs (PWID) are at high risk of severe wounds, invasive infections, and overdoses. To date, there are few data on the bacterial and chemical contaminants PWID are exposed to when using illicitly manufactured fentanyls and stimulants. Methods Previously used injection drug use equipment was recovered in St Louis, Missouri, by harm reduction organizations over a 12-month period. Syringe residue was analyzed for bacterial contaminants by routine culturing followed by whole genome sequencing of single bacterial isolates. Chemical adulterants in syringe residue were identified by liquid chromatography–mass spectrometry. Results Bacteria were cultured from 58.75% of 160 syringes analyzed. Polymicrobial growth was common and was observed in 23.75% of samples. Bacillus cereus was the most common pathogen present and was observed in 20.6% of syringe residues, followed closely by Staphylococcus aureus at 18.8%. One hundred syringes underwent mass spectrometry, which demonstrated that chemical adulterants were common and included caffeine, diphenhydramine, lidocaine, quinine, and xylazine. Conclusions Analysis of syringe residue from discarded drug use equipment demonstrates both chemical and biological contaminants, including medically important pathogens and adulterants. [ABSTRACT FROM AUTHOR]

The fleshy fruit of tomato (Solanum lycopersicum) are climacteric and, as such, ethylene plays a pivotal role in their ripening and quality traits. In this study, a basic helix–loop–helix transcription factor, EMB1444-like, was found to induce the expression of YELLOW-FRUITED TOMATO 1 (YFT1), which encodes the SlEIN2 protein, a key element in the ethylene signaling pathway. Yeast one-hybrid and EMSA analyses revealed that EMB1444-like binds to the E-box motif (CACTTG, –1295 bp to –1290 bp upstream of the ATG start codon) of the YFT1 promoter (pYFT1). Suppression of EMB1444-like expression in tomato lines (sledl) using RNAi reduced ethylene production by lowering the expression of 1-AMINOCYCLOPROPANE-1-CARBOXYLATE SYNTHASE 2/4 (ACS2/4) and ACC OXIDASE1 (ACO1) in a positive feedback loop. sledl tomato also showed differences in numerous quality traits related to fruit ripening, compared with the wild type, such as delayed chromoplast differentiation, a decrease in carotenoid accumulation, and delayed fruit ripening in an ethylene-independent manner, or at least upstream of ripening mediated by YFT1/SlEIN2. This study elucidates the regulatory framework of fruit ripening in tomato, providing information that may be used to breed tomato hybrid cultivars with an optimal balance of shelf-life, durability, and high quality. [ABSTRACT FROM AUTHOR]

Fruit color is a genetic trait and a key factor for consumer acceptability and is therefore receiving increasing importance in several breeding programs. Plant pigments offer plants with a variety of colored organs that attract animals for pollination, favoring seed dispersers and conservation of species. The pigments inside plant cells not only play a light-harvesting role but also provide protection against light damage and exhibit nutritional and ecological value for health and visual pleasure in humans. Tomato (Solanum lycopersicum) is a leading vegetable crop; its fruit color formation is associated with the accumulation of several natural pigments, which include carotenoids in the pericarp, flavonoids in the peel, as well as the breakdown of chlorophyll during fruit ripening. To improve tomato fruit quality, several techniques, such as genetic engineering and genome editing, have been used to alter fruit color and regulate the accumulation of secondary metabolites in related pathways. Recently, clustered regularly interspaced short palindromic repeat (CRISPR)-based systems have been extensively used for genome editing in many crops, including tomatoes, and promising results have been achieved using modified CRISPR systems, including CAS9 (CRISPR/CRISPR-associated-protein) and CRISPR/Cas12a systems. These advanced tools in biotechnology and whole genome sequencing of various tomato species will certainly advance the breeding of tomato fruit color with a high degree of precision. Here, we attempt to summarize the current advancement and effective application of genetic engineering techniques that provide further flexibility for fruit color formation. Furthermore, we have also discussed the challenges and opportunities of genetic engineering and genome editing to improve tomato fruit color. [ABSTRACT FROM AUTHOR]

To create novel tomato (Solanum lycopersicum) germplasm, a wild tomato relative, S. pimpinellifolium (a red-fruited LA1585 accession), was used as the male parent to cross with the yellow-fruited tomato mutant, e9292 (S. lycopersicum). Forty-four morphological characteristics were examined in the present study; 22 S. pimpinellifolium (LA1585) traits and six S. lycopersicum (e9292) traits were dominant in the hybrids (first generation, F1); 10 were intermediate types, and the remaining six resulted in a nonmorphological difference. Ten representative lines were chosen from 1338 line of the second generation of the hybrid (e9292 × LA1585). The ascorbic acid content was higher in three F2 hybrids than in LA1585, which had higher parental levels, as well as the lycopene content in two of the hybrid lines. The fructose and glucose contents were higher in five F2 hybrids compared to e9292, and the sugar/acid value was higher in three hybrid lines. The broad-sense heritability values ranged from 75.06% for total soluble solids to 99.92% for ascorbic acid, and the average value was 92.66% for 15 quality traits. When seedlings were exposed to NaCl and mannitol, the tolerance of high salt concentrations and drought was enhanced in two hybrid lines (F2-266 and F2-299) compared with else tomatoes (e9292, LA1585, F1 hybrids, and F2-332). We created novel tomato germplasm resources with horticulturally desirable quality traits and abiotic stress tolerance, thus offering a methodology for novel tomato germplasm creation and evaluation. [ABSTRACT FROM AUTHOR]

In this study, copper nanoparticles (Cu NPs) were synthesized in an eco-friendly pathway applying Mentha piperita extract as reducing/stabilizing agent. The morphological and physicochemical features of the prepared nanoparticles were determined using several advanced techniques like FE-SEM, EDX, and UV–Vis studies. The as synthesized Cu NPs was explored in the antioxidant and anti-human esophageal cancer tests. The in vitro cytotoxic and anti-esophageal cancer effects of biologically synthesized Cu NPs against human esophageal squamous cell carcinoma (KYSE-270), human caucasian esophageal carcinoma (OE33), and adenocarcinoma of the gastroesophageal junction (ESO26) cancer cell lines were assessed. The anti-esophageal cancer properties of the Cu NPs could significantly remove KYSE-270, OE33, and ESO26 cancer cell lines in a time and concentration-dependent manner by MTT assay. The IC50 of the Cu NPs were 241, 278, and 240 µg/mL against KYSE-270, OE33, and ESO26 cancer cell lines. The antioxidant activity of Cu NPs was determined by DPPH method. The Cu NPs showed the high antioxidant activity according to the IC50 value. It seems that the anti-human esophageal cancer effect of recent nanoparticles is due to their antioxidant effects. [ABSTRACT FROM AUTHOR]

Objectives: Gigantopithecus blacki, the largest hominoid known, is one of the representative Pleistocene mammals in southern China and northern Southeast Asia. Here we investigate the feeding ecology of G. blacki in its core habitat (Guangxi, Southern China) during the early Early Pleistocene, which was the early period in its evolution. Materials and methods: The stable isotopic (C, O) analysis of tooth enamel of the fauna associated with G. blacki (n = 58), including the largest number of G. blacki teeth (n = 12) to date from the Liucheng Gigantopithecus Cave (~2 Ma), Guangxi, China, is undertaken. Results: The δ13C values of Liucheng fauna range from −12.9 to −19.0‰ with an average of −16.1 ± 1.3‰ (n = 58) and the δ18O values range from −4.3 to −9.6‰ with an average of −6.9 ± 1.2‰ (n = 58). The δ13C values of G. blacki range from −15.9‰ to −17.0‰ with an average of −16.5 ± 0.4‰ (n = 12), and the δ18O values vary from −5.9‰ to −7.5‰ with an average of −6.6 ± 0.5‰ (n = 12). Conclusions: The isotopic data show Guangxi was characterized by closed C3 forest and humid climate in the early Early Pleistocene. Niche partitioning is found among G. blacki, Sinomastodon, Ailuropoda and Stegodon, the typical megafauna in South China in the early Early Pleistocene. This could be one of the important factors for them to co‐exist until the Middle Pleistocene. Smallest isotopic variations of G. blacki are found compared with those of contemporary animals, indicating a conservative foraging ecology i.e., limited foraging area and/or narrow dietary flexibility. Furthermore, the more confined foraging ecology of G. blacki is also seen in comparison with fossil and extant large‐bodied primates. However, the unique dietary pattern of G. blacki does not seem to have hindered its survival. The environment in Guangxi during the early Early Pleistocene offered the suitable conditions for G. blacki to become one of the typical species in the faunal assemblages. Highlights: Isotopic results of Gigantopithecus fauna from Liucheng Gigantopithecus Cave, southern China, indicate a paleoecological setting of a C3 dense forest under humid conditions during the early Early Pleistocene.Significant foraging differences are found among G. blacki, giant panda, Stegodon and Sinomastodon.Small isotopic variations of G. blacki provide the first direct evidence of conservative foraging ecology (limited foraging area and/or confined dietary flexibility) in its core habitat during its early evolution.Narrower dietary breadth of G. blacki during the early Early Pleistocene might relate to its large body size in comparison with other modern and fossil large‐bodied primates. [ABSTRACT FROM AUTHOR]

Rice blast, caused by the fungus Magnaporthe oryzae , is the most devastating disease affecting rice production. Identification of protein–protein interactions (PPIs) is a critical step toward understanding the molecular mechanisms underlying resistance to blast fungus in rice. In this study, we presented a computational framework for predicting plant–pathogen PPIs based on structural information. Compared with the sequence-based methods, the structure-based approach showed to be more powerful in discovering new PPIs between plants and pathogens. Using the structure-based method, we generated a global PPI network consisted of 2,018 interacting protein pairs involving 1,344 rice proteins and 418 blast fungus proteins. The network analysis showed that blast resistance genes were enriched in the PPI network. The network-based prediction enabled systematic discovery of new blast resistance genes in rice. The network provided a global map to help accelerate the identification of blast resistance genes and advance our understanding of plant–pathogen interactions. [ABSTRACT FROM AUTHOR]

Fruit quality in most fleshy fruit crops is fundamentally linked to ripening-associated traits, including changes in colour. In many climacteric fruits, including tomato (Solanum lycopersicum), the phytohormone ethylene plays a key role in regulating ripening. Previous map-based cloning of YELLOW FRUITED-TOMATO 1 (YFT1) revealed that it encodes the EIN2 protein, a core component in ethylene signal transduction. A YFT1 allele with a genetic lesion was found to be down-regulated in the yft1 tomato mutant that has a yellow fruit phenotype and perturbed ethylene signalling. Based on bioinformatic analysis, yeast one hybrid assays and electrophoretic mobility shift assays, we report that transcription factor WRKY32 regulates tomato fruit colour formation. WRKY32 binds to W-box and W-box-like motifs in the regulatory region of the YFT1 promoter and induces its expression. In tomato fruits of WRKY32-RNAi generated lines, ethylene signalling was reduced, leading to a suppression in ethylene emission, a delay in chromoplast development, decreased carotenoid accumulation, and a yellow fruit phenotype. These results provide new insights into the regulatory networks that govern tomato fruit colour formation via ethylene signal transduction. [ABSTRACT FROM AUTHOR]

Objectives: We aimed to investigate molar enamel development in fossil orangutans from Guangxi and shed light on the evolution of Asian great apes. Materials and methods: We collected 32 fossil orangutan molars, most of which were from Guangxi apothecaries and the Guangxi Daxin Heidong cave, and prepared histological sections of each molar. We then characterized aspects of dental development, including long period line periodicity, number of Retzius lines and lateral enamel formation time, cuspal enamel thickness, and enamel formation time. Results: The long period line periodicity in fossil orangutans ranged from 9 to 10 days (mean, 9.09 days). The molar lateral enamel formation time ranged from 1.48 to 3.17 years (540–1,152 days). Cuspal enamel thickness in fossil orangutan molars ranged from 949 to 2,535 μm, and cuspal enamel formation time ranged from 0.64 to 1.87 years. Molar enamel formation time of fossil orangutans ranged from 2.47 to 4.67 years. Discussion Long‐period line periodicity of fossil orangutans from Guangxi was within the variation range of extant orangutans, and the average long period line periodicity (9.09 days) of fossil orangutans from Guangxi in this study was lower than the values for extant orangutans (9.5 days) and fossil orangutans (10.9 days) from Sumatra and Vietnam. Orangutan enamel thickness may have gradually decreased from the Middle Pleistocene to Holocene. Crown formation time of fossil orangutans was slightly longer than that of extant orangutans, and the M1 emergence age of fossil orangutans from Guangxi was about 4–6 years. These findings might indicate the regional difference or evolutionary changes in orangutans since Pleistocene. Dental development of the Guangxi fossil orangutans were more similar to that of Asian Miocene apes, suggesting the closer evolutionary relationship of orangutans to Miocene Asian fossil apes. [ABSTRACT FROM AUTHOR]

Rice ( Oryza sativa) is one of the most important staple foods for more than half of the global population. Many rice traits are quantitative, complex and controlled by multiple interacting genes. Thus, a full understanding of genetic relationships will be critical to systematically identify genes controlling agronomic traits. We developed a genome-wide rice protein-protein interaction network (RicePPINet, ) using machine learning with structural relationship and functional information. Rice PPINet contained 708 819 predicted interactions for 16 895 non-transposable element related proteins. The power of the network for discovering novel protein interactions was demonstrated through comparison with other publicly available protein-protein interaction ( PPI) prediction methods, and by experimentally determined PPI data sets. Furthermore, global analysis of domain-mediated interactions revealed Rice PPINet accurately reflects PPIs at the domain level. Our studies showed the efficiency of the Rice PPINet-based method in prioritizing candidate genes involved in complex agronomic traits, such as disease resistance and drought tolerance, was approximately 2-11 times better than random prediction. Rice PPINet provides an expanded landscape of computational interactome for the genetic dissection of agronomically important traits in rice. [ABSTRACT FROM AUTHOR]

Objectives The present study investigated the distribution of perikymata on anterior teeth of Miocene Lufengpithecus lufengensis so as to broaden the comparative data of developmental variation within and among hominoids. We also compared perikymata-spacing pattern of Lufengpithecus lufengensis with hominins and extant African great apes to understand the implication of dental development. Materials and methods A total of 30 anterior teeth (including 6 I1, 10 I2, and 14 C) of Lufengpithecus lufengensis were examined using a scanning electron microscope and Keyence VHX-600EOS digital microscope to document the number and distribution of perikymata on their labial surfaces. The labial crown height of each tooth was divided into 10 equal deciles and the total perikymata number in each decile was recorded. The mean number of perikymata per millimeter was then calculated for each decile. SPSS statistical software was used to perform analyses of these data, including t-tests for sexual dimorphism and plots showing the perikymata distribution in Lufengpithecus lufengensis. Results Perikymata counts of Lufengpithecus lufengensis in the first three deciles are fewer than the remaining deciles, and with perikymata becoming increasingly more closely packed as growth progresses from cusp to cervix, but decrease in density in the cervical decile. Besides, total labial perikymata counts of canines tend to display very significant sexual dimorphism. Discussion Lufengpithecus lufengensis anterior teeth are more similar in their distribution of labial perikymata to those of Australopithecus than those of other hominins and extant African great apes from previous studies. [ABSTRACT FROM AUTHOR]

Thymosin β4 (Tβ4) is a peptide that is known to play important roles in protection, regeneration, and remodeling of injured tissues in humans, and that shows great promise in a range of clinical applications. However, current strategies to Tβ4 are insufficient to meet growing demand and have a number of limitations. In this current study we investigated whether expression of recombinant Tβ4 in plants, specifically in tobacco (Nicotiana tabacum) leaves, represents an effective approach. To address this question, a 168 bp Tβ4 gene optimized for tobacco codon usage bias was constitutively expressed in tobacco as a 4-unit repeat concatemer, fused to a polyhistidine tag. Quantitative polymerase chain reaction and Western blot analyses were used to verify 4×Tβ4 expression in 14 transgenic tobacco lines and enzyme-linked immunosorbent assay analysis indicated 4×Tβ4 protein concentrations as high as 3 μg/g of fresh weight in the leaves. We observed that direct administration of tobacco-derived Tβ4 was more effective than Tβ4 either obtained commercially or derived from expression in Escherichia coli at promoting splenocyte proliferation in vitro and wound healing in mice through an endothelial migration assay. This study provides new insights into the development of plant-derived therapeutic proteins and their application by direct administration. [ABSTRACT FROM AUTHOR]

The plant cuticle is thought to be a critical evolutionary adaptation that allowed the first plants to colonize land, because of its key roles in regulating plant water status and providing protection from biotic and abiotic stresses. Much has been learned about cuticle composition and structure through genetic and biochemical studies of angiosperms, as well as underlying genetic pathways, but little is known about the cuticles of early diverging plant lineages. Here, we demonstrate that the moss Physcomitrella patens , an extant relative of the earliest terrestrial plants, has a cuticle that is analogous in both structure and chemical composition to those of angiosperms. To test whether the underlying cuticle biosynthetic pathways were also shared among distant plant lineages, we generated a genetic knockout of the moss ATP binding cassette subfamily G (ABCG) transporter Pp-ABCG7, a putative ortholog of Arabidopsis thaliana   ABCG transporters involved in cuticle precursor trafficking. We show that this mutant is severely deficient in cuticular wax accumulation and has a reduced tolerance of desiccation stress compared with the wild type. This work provides evidence that the cuticle was an adaptive feature present in the first terrestrial plants and that the genes involved in their formation have been functionally conserved for over 450 million years. [ABSTRACT FROM AUTHOR]

Hepatocellular carcinoma ( HCC) is a common clinical primary malignant tumor; however, efficient drugs for the treatment of HCC are still lacking at the present time. To develop a new approach for liver cancer therapy, we designed a chimeric gene ( his-HR) encoding a single-chain variable fragment of human HAb25 (h Hsc Fv) fused to a cytotoxic ribonuclease from Rana catesbeiana ( RC- RNase) and expressed the corresponding fusion protein in transgenic tobacco ( Nicotiana tabacum). Eleven positive transgenic plant lines were identified from 204 regenerated tobacco plants by PCR and Southern blot analysis, and the immunocompetence of the recombinant his- HR protein was confirmed by Western blotting. The expression levels of his- HR protein ranged from 0.75 to 1.99 µg/g in the fresh tobacco leaves. To characterize the bifunction of the expressed his- HR protein in tobacco, binding specificity and cell toxicity to several cell lines were examined by the indirect immunocytochemical streptavidin-biotin complex method and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2 H-tetrazolium bromide assay. Data indicated that the his- HR protein had stronger specific binding affinity to Hep G2 (human liver HCC cell line) than to the other tumor cell lines and normal liver cell line, and the capacity to kill the HCC cell lines SMMC7721 and Hep G2 with an half maximal inhibiting concentration of 2.0 and 2.4 nM, respectively. The results suggest that recombinant bifunctional his- HR protein derived from transgenic plants may provide a novel strategy to treat HCC in the future. [ABSTRACT FROM AUTHOR]

Abstract: Vitamin E has been found to be associated with an important antioxidant property in mammals and plants. In photosynthetic organisms, the enzyme 4-hydroxyphenylpyruvate dioxygenase (HPPD; E.C. 1.13.11.27) plays an important role in the vitamin E biosynthetic pathway. The full-length cDNA encoding HPPD was isolated from Lactuca sativa L. by rapid amplification of cDNA ends (RACE). The cDNA, designated as LsHPPD, was 1743 base pairs (bp) long containing an open reading frame (ORF) of 1338bp encoding a protein of 446 amino acids. Sequence analysis indicated that LsHPPD shared high identity with HPPD from Medicago truncatula L. Real-time fluorescent quantitative PCR (qPCR) analysis revealed that LsHPPD was preferentially expressed in mature leaves compared with other tissues and that the LsHPPD expression was sensitive to high light and drought stress treatments. Transient expression of LsHPPD via agroinfiltration resulted in 12-fold increase in LsHPPD mRNA expression level and 4-fold enhancement in α-tocopherol content compared with the negative control. A decrease in chlorophyll content and inhibition of photosystem II were observed during stress treatments and agroinfiltration. [Copyright &y& Elsevier]

Genetic modification can be an effective strategy for improving the agronomic traits of tomato (Solanum lycopersicum) to meet demands for yield, quality, functional components, and stress tolerance. However, limited numbers of available tissue-specific promoters represent a bottleneck for the design and production of transgenic plants. In the current study, a total of 25 unigenes were collected from an RNA-sequence dataset based on their annotation as being exclusively expressed in five type of tissues of tomato pericarp (outer and inner epidermis, collenchyma, parenchyma, and vascular tissues), and every five unigenes, was respectively selected from each tissue based on transcription expression. The 3-kb 5′ upstream region of each unigene was identified from the tomato genome sequence (SL2.50) using annotated unigene sequences, and the promoter sequences were further analyzed. The results showed an enrichment in T/A (T/A > 70%) in the promoter regions. A total of 15 putative tissue-/organ-specific promoters were identified and analyzed by real-time (RT) quantitative (q) PCR analysis, of which six demonstrated stronger activity than widely used tissue-specific tomato promoters. These results demonstrate how high spatiotemporal and high throughput gene expression data can provide a powerful means of identifying spatially targeted promoters in plants. [ABSTRACT FROM AUTHOR]