Transcriptome and anatomical studies reveal alterations in leaf thickness under long-term drought stress in tobacco.

Drought is one of the foremost environmental factors that limit the growth of plants. Leaf thickness (LT) is an important quantitative trait in plant physiology. The experiment was carried out in a growth room and the plants were divided into two groups such as well-watered and drought-stressed. This work investigated leaf growth in terms of leaf surface growth and expansion rate, leaf stomata traits, LT, anticlinal growth, and leaf cell layers. The results showed that the leaf area and leaf surface expansion rate were decreased by drought stress (DS). Similarly, LT, anticlinal expansion rate, palisade and spongy tissue thickness, and their related expansion rates were also decreased at different days' time points (DTP) of DS. However, a steady increase was observed in the aforementioned parameters after 12 DTP of DS. The stomatal density increased while stomata size decreased at 3 DTP and 12 DTP (low leaf water potential and relative leaf water content at these time points) and vice versa at 24 DTP compared with the well-watered plants indicating adaptations in these traits in response to DS, and thus the leaf water status played a role in the regulation of leaf stomata traits. The cell length decreased in the upper epidermis, palisade and spongy tissues by DS up to 12 DTP led to lower LT while an increase was observed after 12 DTP that resulted in higher LT. The increase in the LT was supported by the upregulation of starch and sucrose metabolism, glycerolipid metabolism, protein processing in endoplasmic reticulum pathways at 18 DTP along with the differentially expressed genes induced that were related to cell wall remodeling (cellulose, expansin, xyloglucans) and cell expansion (auxin response factors and aquaporin). The results explain the response of leaf thickness to drought stress and show alterations in LT and leaf stomatal traits. This study might serve as a valuable source of gene information for functional studies and provide a theoretical basis to understand leaf growth in terms of leaf anatomy and leaf stomatal traits under drought stress.

Application of 2,4-Epibrassinolide Improves Drought Tolerance in Tobacco through Physiological and Biochemical Mechanisms.

Drought stress is a major abiotic stress that hinders plant growth and development. Brassinosteroids (BR), including 2,4-epibrassinolide (EBR), play important roles in plant growth, development, and responses to abiotic stresses, including drought stress. This work investigates exogenous EBR application roles in improving drought tolerance in tobacco. Tobacco plants were divided into three groups: WW (well-watered), DS (drought stress), and DSB (drought stress + 0.05 mM EBR). The results revealed that DS decreased the leaf thickness (LT), whereas EBR application upregulated genes related to cell expansion, which were induced by the BR (DWF4, HERK2, and BZR1) and IAA (ARF9, ARF6, PIN1, SAUR19, and ABP1) signaling pathway. This promoted LT by 28%, increasing plant adaptation. Furthermore, EBR application improved SOD (22%), POD (11%), and CAT (5%) enzyme activities and their related genes expression (FeSOD, POD, and CAT) along with a higher accumulation of osmoregulatory substances such as proline (29%) and soluble sugars (14%) under DS and conferred drought tolerance. Finally, EBR application augmented the auxin (IAA) (21%) and brassinolide (131%) contents and upregulated genes related to drought tolerance induced by the BR (BRL3 and BZR2) and IAA (YUCCA6, SAUR32, and IAA26) signaling pathways. These results suggest that it could play an important role in improving mechanisms of drought tolerance in tobacco.

Oral transmucosal absorption of iodine after intraoral preparation with povidone-iodine prior to oral surgeries: a randomized controlled study in 12 male patients.

INTRODUCTION: Absorption of iodine through skin and an increased incidence of thyroid disorders due to iodophor exposure are reported. However, the risk of oral transmucosal absorption of povidone-iodine after intraoral preparation is not clear. OBJECTIVE: To investigate the possibility of oral transmucosal absorption of povidone-iodine after intraoral preparation and its effect on thyroxine level in blood. PATIENTS AND METHODS: A randomized controlled study was carried out in the department of oral and maxillofacial surgery. Twenty- to 40-year-old healthy male adults planning to receive oral surgery under general anesthesia were enrolled. The study group received povidone-iodine irrigation of oral cavity for 3 min as intraoral preparation before operation. The control group received chlorhexidine gluconate irrigation of oral cavity for 3 min as intraoral preparation before operation. Iodine levels in blood and urine, and thyroxine levels in blood were tested and compared, before and after operation. RESULT(S): In total, 24 patients were included and analyzed finally. We found that after intraoral preparation with povidone-iodine, serum iodine level increased significantly to 2-3 times the pre-operation level in 15-30 min, and the urinary iodine level increased to 5 times the pre-operation level on the first day after operation. Iodine levels in blood and urine decreased significantly on the third day after operation but still significantly greater than the pre-operation levels. However, thyroxine levels were not altered accordingly. CONCLUSION(S): Oral transmucosal absorption of iodine is observed when povidone-iodine is used for intraoral preparation in healthy male adults, though the free thyroxine in blood is not affected accordingly. CLINICAL RELEVANCE: Povidone-iodine is commonly used as an antiseptic in oral surgery and dental clinics. Based on our findings that iodine levels in blood and urine may elevate significantly after intraoral preparation with povidone-iodine prior to oral surgeries, care must be taken for individuals when excess iodine intakes can endanger the safety of the patient. REGISTRATION INFORMATION: Name of the trial registry: The Chinese Clinical Trial Registry ( http://www.chictr.org.cn/ ). Registration number: ChiCTR2100042028.

Identification and characterization of long noncoding RNAs and mRNAs expression profiles related to postnatal liver maturation of breeder roosters using Ribo-zero RNA sequencing.

BACKGROUND: The liver is mainly hematopoietic in the embryo, and converts into a major metabolic organ in the adult. Therefore, it is intensively remodeled after birth to adapt and perform adult functions. Long non-coding RNAs (lncRNAs) are involved in organ development and cell differentiation, likely they have potential roles in regulating postnatal liver development. Herein, in order to understand the roles of lncRNAs in postnatal liver maturation, we analyzed the lncRNAs and mRNAs expression profiles in immature and mature livers from one-day-old and adult (40 weeks of age) breeder roosters by Ribo-Zero RNA-Sequencing. RESULTS: Around 21,939 protein-coding genes and 2220 predicted lncRNAs were expressed in livers of breeder roosters. Compared to protein-coding genes, the identified chicken lncRNAs shared fewer exons, shorter transcript length, and significantly lower expression levels. Notably, in comparison between the livers of newborn and adult breeder roosters, a total of 1570 mRNAs and 214 lncRNAs were differentially expressed with the criteria of log2fold change > 1 or < - 1 and P values < 0.05, which were validated by qPCR using randomly selected five mRNAs and five lncRNAs. Further GO and KEGG analyses have revealed that the differentially expressed mRNAs were involved in the hepatic metabolic and immune functional changes, as well as some biological processes and pathways including cell proliferation, apoptotic and cell cycle that are implicated in the development of liver. We also investigated the cis- and trans- regulatory effects of differentially expressed lncRNAs on its target genes. GO and KEGG analyses indicated that these lncRNAs had their neighbor protein coding genes and trans-regulated genes associated with adapting of adult hepatic functions, as well as some pathways involved in liver development, such as cell cycle pathway, Notch signaling pathway, Hedgehog signaling pathway, and Wnt signaling pathway. CONCLUSIONS: This study provides a catalog of mRNAs and lncRNAs related to postnatal liver maturation of chicken, and will contribute to a fuller understanding of biological processes or signaling pathways involved in significant functional transition during postnatal liver development that differentially expressed genes and lncRNAs could take part in.

Effect of dietary Astragalus Polysaccharide supplements on testicular miRNA expression profiles and enzymatic changes of breeder cocks.

Astragalus Polysaccharide (APS) is an important feed additive due to its immunomodulatory functions. Previous studies have proven that miRNAs play important roles in posttranscriptional gene regulation. Our goals were to identify differentially expressed miRNAs in testes in responses to APS dietary supplements and to find the effects of APS on breeder cock testes. We measured several enzymatic activities in testes and sperm samples and further generated miRNA expression profiles of testes from breeder cocks fed with control diets and extra APS. As a result, we found APS could increase testicular functional activities of marker enzymes. Meanwhile, there were 16 up-regulated and 17 down-regulated miRNAs in APS group, compared with the control group meeting the criteria of P-values < 0.05. Meanwhile, twelve differentially expressed miRNAs were validated by Mir-XTM miRNA RT-qPCR. Further GO and KEGG analyses of target genes for differentially expressed miRNAs revealed that some miRNAs may be involved in testicular nutrient metabolisms and NK cell mediated cytotoxicity pathway. Moreover, the effect of dietary APS supplements on NK cell mediated cytotoxicity pathway was also validated by RT-qPCR. Our results provided a novel insight into the effect of dietary APS supplements on testicular miRNA expression profiles and enzymatic changes of breeder cocks.

Combined Use of Facial Osteoplasty and Orthognathic Surgery for Treatment of Dentofacial Deformities.

PURPOSE: Orthognathic surgery is an efficient procedure for cosmetic and functional aims. However, when functional improvement is achieved by mandibular or maxillary operations, additional esthetic corrections may be imperative for some patients. This study aims to introduce our primary practice of simultaneous facial bone contouring and orthognathic surgery for esthetic reasons. PATIENTS AND METHODS: Ten patients with dentofacial deformities as well as a prominent angle, asymmetric deformities, or a high zygoma and zygomatic arch were recruited from West China Hospital of Stomatology, Sichuan University (Chengdu, China), between January 1, 2014, and July 31, 2015. Traditional orthognathic surgical procedures such as bilateral sagittal split osteotomy and Le Fort I osteotomy combined with facial osteoplasty including mandibular angle ostectomy, outer cortex ostectomy of the mandibular angle, and zygoma and zygomatic arch reduction were performed. Radiographs and medical photographs were taken before and after surgery to compare the effectiveness of the combined use of facial osteoplasty and orthognathic surgery. RESULTS: All patients had an uneventful postoperative recovery, with no signs of infection, jaw displacement, or osteonecrosis. Radiographs taken 1 week after surgery and pictures of the facial profile and occlusion taken 6 months after surgery showed satisfactory esthetic outcomes. All patients were satisfied with the functional and cosmetic results. CONCLUSIONS: This study indicated the clinical feasibility of simultaneous facial bone contouring and orthognathic surgery for the treatment of dentofacial deformities. Simultaneous facial bone contouring seems to be an alternative procedure in addition to conventional orthognathic surgery for cosmetic aims in certain patients.

Changes in DNA Methylation and Chromatin Structure of Pro-inflammatory Cytokines Stimulated by LPS in Broiler Peripheral Blood Mononuclear Cells.

The pro-inflammatory cytokines IL-1ß, IL-6, and tumor necrosis factor (TNF)-α mediate inflammation, which is a protective response by body to ensure removal of detrimental stimuli, as well as a healing process for repairing damaged tissue. The overproduction of pro-inflammatory cytokines can induce autoimmune diseases and can be fatal. The aim of this study was to investigate epigenetic mechanisms in the regulation of pro-inflammatory cytokines expression after lipopolysaccharide (LPS) stimulation of broiler peripheral blood mononuclear cells (PBMC). Gene expression, promoter DNA methylation, and chromatin accessibility of pro-inflammatory cytokines in untreated and LPS-treated PBMC were compared. The expression of epigenetic enzymes DNA methyltransferase (DNMT) 1, histone deacetylase (HDAC), and histone acetylase (HAT) were measured after LPS stimulation. The results showed the activated gene expression of pro-inflammatory cytokines in broiler PBMC stimulated 3 h by LPS. The demethylation of IL-6 gene - 302 and -264 cytosine-guanine (CpG) sites, as well as TNF-α gene -371 CpG site, occurred after LPS treatment (P < 0.05), whereas the methylaiton pattern in the IL-1ß gene promoter region was not affected. Otherwise, LPS stimulation relaxed the chromatin structure at IL-1ß and IL-6 promoter (P < 0.05). The lower expression of DNMT1 and HDAC2, and higher expression of p300-CBP-associated factor and tat-interaction protein-60, were detected in response to LPS (P < 0.05). Our data indicated that after LPS stimulation for 3 h, IL-1ß and IL-6 promoter are remodeled into an accessible structure, and the IL-6 and TNF-α promoter are demethylated at special sites, which possible impact the mRNA expression of pro-inflammatory cytokines.