Group IIc Research Articles

The ultrastructural changes in the adult rat ovary after administration of copper oxide nanoparticles and the possible ameliorative influence of selenium

ABSTRACT There is an important concern about the potential health and environmental risks that may develop due to exposure to copper oxide nanoparticles (CuO-NPs). Selenium is an essential trace element. It supports the expression of a variety of selenoproteins. The present study was designed to study the ultrastructural and biochemical changes in the adult rat ovary after oral administration of CuO-NPs and to assess the possible ameliorative influence of Selenium. Sixty adult female albino rats were divided in two major groups: Group I and Group II. Group I was further subdivided into three groups: Group IA (control), Group IB: received a single high dose of 2000 mg/kg CuO-NPs, Group IC: received selenium (0.5 mg/kg), five days before giving a single high dose of CuO-NPs (2000 mg/kg). Thereafter, given selenium for 14 days. Group II was subdivided into three groups: Group IIA (control), Group IIB: received a small dose of 300 mg/kg of CuO-NPs for 28 days, Group IIC: received selenium (0.5 mg/kg), five days before starting concomitant administration of CuO-NPs (300 mg/kg) and Selenium (0.5 mg/kg) for 28 days. Damage of the ovarian ultrastructural features, increased MDA levels, and decreased serum estrogen and progesterone hormones levels were detected in group IB and group IIB. Group IC and group IIC showed improvement of ovarian ultrastructural, decreased MDA levels, and increased serum estrogen and progesterone hormones levels as compared to group IB and group IIB indicating that Selenium could decrease the damage induced by CuO-NPs in the adult rat ovaries.

Impact of Copper Oxide Nanoparticles on Adult Rat Ovary and Possible Protective Effect of Selenium: A Histological and Immunohistochemical Study

Copper oxide nanoparticles are thought to be an important transition metal oxide due to their interesting properties. Selenium is an important structural component of numerous enzymes, including glutathione peroxidase (GPx), thioredoxin reductase, as well as deiodinases.The present work was undertaken to evaluate the histological and immunohistochemical alterations in the adult rat ovary after oral administration of CuO-NPs, and to assess the potential ameliorative impact of selenium. Sixty adult female albino rats were equally divided in two major groups: group I and group II. Group I was further divided into three groups: control group IA, group IB: given 2000 mg/kg CuO-NPs once, group IC: received selenium (0.5 mg/kg), five days before giving one high dose of 2000 mg/kg CuO-NPs. Thereafter, given selenium for 14 days. group II was subdivided into three groups: group IIA (control), group IIB: received 300 mg/kg/dayof CuO-NPs for 28 days, group IIC: received selenium (0.5 mg/kg), five days before starting concomitant administration of 300 mg/kg CuO-NPs and 0.5 mg/kg selenium for 28 days. Both group IB and group IIB showed signs of follicle and corpora lutea degeneration and revealed increased numbers of apoptotic bodies and atretic follicles. Increased reactions for caspase 3 and decreased reactions for BCL2 were also demonstrated. Apoptosis was greater in group IB. Group IC and group IIC showed apparent improvement of the histological structure, decreased numbers of apoptotic bodies (by 34.3% for the acute study and 38% for the chronic study)，decreased atretic follicles (by 37.5% for the acute study and 34.78% for the chronic study), decreased reactions for caspase 3 and increased reactions for BCL2 as compared to groups IB and IIB respectively.

Genome-wide analysis of the WRKY gene family and their response to low-temperature stress in elephant grass.

Elephant grass (Cenchrus purpureus) is a perennial forage grass characterized by tall plants, high biomass and wide adaptability. Low-temperature stress severely limits elephant grass biomass and geographic distribution. WRKY is one of the largest families of plant-specific transcription factors and plays important roles in plant resistance to low-temperature. However, the understanding of the WRKY family in grasses is limited. In this study, we conducted a genome-wide characterization of WRKY proteins in elephant grass, including gene structure, phylogeny, expression, conserved motif organization, and functional annotation, to identify key CpWRKY candidates involved in cold tolerance. In this study, a total of 176 WRKY genes were identified in elephant grass. It was found that 172 were unevenly distributed across its 14 chromosomes, while the remaining 4 genes were not anchored to any chromosome. The genes were classified into three groups based on their WRKY conserved domains and zinc finger motifs. There were 12, 8, 19, 27, 12, 18 and 80 CpWRKYs belonging to group I, group IIa, group IIb, group IIc, group IId, group IIe and group III, respectively. We hypothesized that the ancient subgroup IIc WRKY gene is the ancestor of all WRKY genes in elephant grass. Most CpWRKYs in the same group have similar structure and motif composition. A total of 169 duplicate gene pairs were identified, suggesting that segmental duplication might have contributed to the expansion of the CpWRKY gene family. Ka/Ks analysis revealed that most of the CpWRKYs were subjected to purifying selection during the evolution. It was also found that six genes (CpWRKY51, CpWRKY81, CpWRKY100, CpWRKY101, CpWRKY140 and CpWRKY143) exhibited higher expression in roots compare to leaves, and were significantly induced by low temperature stress. Among them, CpWRKY81 had the highest expression under low-temperature stress, and its over-expression significantly enhanced the cold tolerance in yeast. In this study, we characterized WRKY genes in elephant grass and further investigated their physicochemical properties, evolution, and expression patterns under low-temperature stress. This research provides valuable resources for identifying key CpWRKY genes that contribute to cold tolerance in elephant grass.

Overexpression of BnaA10.WRKY75 Decreases Cadmium and Salt Tolerance via Increasing ROS Accumulation in Arabidopsis and Brassica napus L.

Soil is indispensable for agricultural production but has been seriously polluted by cadmium and salt in recent years. Many crops are suffering from this, including rapeseed, the third largest global oilseed crop. However, genes simultaneously related to both cadmium and salt stress have not been extensively reported yet. In this study, BnaA10.WRKY75 was screened from previous RNA-seq data related to cadmium and salt stress and further analyses including sequence comparison, GUS staining, transformation and qRT-PCR were conducted to confirm its function. GUS staining and qRT-PCR results indicated BnaA10.WRKY75 was induced by CdCl2 and NaCl treatment. Sequence analysis suggested BnaA10.WRKY75 belongs to Group IIc of the WRKY gene family and transient expression assay showed it was a nuclear localized transcription factor. BnaA10.WRKY75-overexpressing Arabidopsis and rapeseed plants accumulated more H2O2 and O2- and were more sensitive to CdCl2 and NaCl treatment compared with untransformed plants, which may be caused by the downregulation of BnaC03.CAT2. Our study reported that BnaA10.WRKY75 increases sensitivity to cadmium and salt stress by disrupting the balance of reactive oxygen species both in Arabidopsis and rapeseed. The results support the further understanding of the mechanisms underlying cadmium and salt tolerance and provide BnaA10.WRKY75 as a valuable gene for rapeseed abiotic stress breeding.

Evaluation of serum levels of sestrin 2 and betatrophin in type 2 diabetic patients with diabetic nephropathy

BackgroundDiabetic kidney disease (DKD) is one of the most serious microvascular complications of diabetes mellitus (DM) and the leading cause of chronic kidney disease (CKD) worldwide. Since obesity and type 2 DM (T2DM) are considered as inflammatory conditions, thus reducing their accompanied systemic inflammation may lessen their complications. Sestrin 2 belongs to a group of stress induced proteins which are produced in response to oxidative stress, inflammation and DNA damage. Betatrophin; a hormone that stimulates the growth, proliferation and mass expansion of pancreatic beta-cells and improves glucose tolerance. The objective of the study was to evaluate levels of serum Sestrin 2 and betatrophin in patients with different stages of diabetic nephropathy (DN)) and compare results with healthy control.MethodsThis cross sectional study was carried out on 60 patients above 18 years old, recruited from Tanta University hospitals out patients clinics and 20 apparently healthy individuals of matched sex and age as a control group. Participants were divided into two groups: group I: 20 normal subjects as control group and group II: 60 patients with type 2 DM,. further subdivided in to three equal groups: group 1IIA(20 patients) with normo-albuminuria (ACR < 30 mg/g), group IIB (20 patients) with micro albuminuria (ACR = 30 to 300 mg/g) and group IIC (20 patients) with macro albuminuria (ACR > 300 mg/g). They were subjected to detailed history taking, careful clinical examination and laboratory investigations including blood urea, serum creatinine, estimated glomerular filtration rate (eGFR), urinary albumin creatinine ratio, and specific laboratory tests for Sestrin 2 and Betatrophin by using ELISA technique.ResultsSerum Sestrin 2 significantly decreased, while serum betatrophin level significantly increased in macroalbuminuric group compared to control and other 2 diabetic groups (P value < 0.05). The cut off value of serum sestrin 2 was 0.98 ng/ml with sensitivity 99%, specificity 66% while the cut off value of serum betatrophin was > 98.25 ng/ml with sensitivity 98%, specificity 82%. Serum betatrophin positively correlated with age, fasting, 2 h postprandial, BMI, triglyceride, total cholesterol, serum creatinine, blood urea, UACR, and negatively correlated with eGFR and serum albumin. Serum Sestrin 2 positively correlated with serum albumin. BMI, serum urea, UACR and serum albumin. Serum betatrophin are found to be risk factors or predictors for diabetic nephropathy.ConclusionsPatients with DN, particularly the macroalbuminuria group, had a significant increase in betatrophin levels and a significant decrease in serum Sestrin 2 level. The function of Sestrin 2 is compromised in DN, and restoring it can reverse a series of molecular alterations with subsequent improvement of the renal functions, albuminuria and structural damage.

Genome‐wide study of OsWRKY gene in Oryza sativa subsp. japonica

AbstractWRKY transcription factor (TF) family is well known to govern essential physiological functioning as well as regulate plant response to biotic and abiotic stress. In this study, we have identified 108 OsWRKY genes in the genome of Oryza sativa subsp. japonica, using the updated genomic data from the Rice Annotation Project Database and Oryzabase, which were further used to conduct the phylogenetic study, motif analysis, gene structure analysis, chromosomal mapping, and prediction of sub‐cellular localization. The multiple sequence alignment OsWRKY proteins revealed the presence of nine different types of alterations in the conserved heptapeptide sequence WRKYGQK associated with 19 OsWRKY genes. Physiochemical analysis discloses the hydrophobic amino acid‐rich, thermally stable, and polar nature of OsWRKY proteins. These genes were noted as highly conserved between the two cultivated sub‐species of Oryza sativa, that is Indica and japonica type. Additionally, from motif analysis, we have found a new motif, which was categorized as hAT family C‐terminal dimerization region associated with four members of group IIc. We have identified 21 stress‐responsive OsWRKY genes, and their significance to the different biotic and abiotic stress‐mediated cascades was further evaluated by analysing 1500 kb upstream sequences and this disclosed the presence of important phytohormone‐responsive cis‐elements in the OsWRKY gene, suggesting its direct involvement in defence against a wide range of external stressors and these 21 OsWRKY genes are tentatively listed as possible candidates for more study.

Assessment of the Additive Effect of Remineralizing Agents in Combination With Fluoride Releasing Adhesives in the Prevention of Enamel Decalcification in Orthodontic Patients: An In Vitro Study.

Introduction Incorporation of remineralizing agents with fluoride-releasing bracket adhesives may prevent the development of white spot lesions (WSL) or reverse the established WSL in patients undergoing fixed orthodontic treatment. We aimed to find out how effectively casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) and fluoride varnish (FV) can remineralize teeth when mixed with fluoride-releasing orthodontic adhesive. Materials and methods We randomly assigned a total of 60 premolar teeth, therapeutically extracted for orthodontic purposes, into two equal groups. Group I (n = 30) utilized fluoride-releasing adhesive (FR), and Group II (n = 30) bonded with non-fluoride adhesive (NFR). Based on the applied remineralizing agent, we further divided each of the two groups into three equal subgroups of 10: Group IA (FR+FV), Group IB (FR+CPP-ACP), Group IC (control-only FR), Group IIA (NFR+FV), Group IIB (NFR+CPP-ACP), and Group IIC (control-only NFR). Following bonding procedures, all the samples underwent pH cycling for 28 days, where the enamel samples were immersed in 20 ml of demineralizing solution for three hours, followed by immersion in 30 ml of remineralizing solution for 17 hours. The samples were analyzed for shear bond strength (SBS) on a universal testing machine and hardness values (HV) by the Vickers microhardness test (VMT) using the indentation method. We also evaluated the adhesive remnant index (ARI) scores to determine the site of bracket failure. Statistical analysis The shear bond strength (SBS) and hardness value (HV) were expressed as the mean, standard deviation (SD), and median for each subgroup. We used the non-parametric Kruskal-Wallis test to analyze the SBS and HV, followed by the Dunn-Bonferroni test for intra-pair differences. The ARI score was expressed as the frequency of the percentage distribution, and the difference in the distribution of ARI scores between the groups was assessed by the Cochran chi-square test. The probability (p) value equal to or less than 0.05 was considered statistically significant. Results The results show that Group IB, bonded with a fluoride-releasing adhesive and a CPP-ACP remineralizing agent surface treatment, has the highest HV of300.23 units. Group IIC (only NFR) has the lowest hardness of153.3 units, which is statistically significant (p < 0.001). However, the ARI scores are not statistically significant between the groups tested. Conclusion The bond strength of the adhesive and the surface hardness of the enamel increased with the addition of fluoride varnish and CPP-ACP to both the fluoride-releasing and non-fluoride-releasing adhesives.

Overexpression of a pearl millet WRKY transcription factor gene, PgWRKY74, in Arabidopsis retards shoot growth under dehydration and salinity-stressed conditions

Pearl millet (Cenchrus americanus) is a cereal crop that can tolerate high temperatures, drought, and low-fertility conditions where other crops lose productivity. However, genes regulating this ability are largely unknown. Transcription factors (TFs) regulate transcription of their target genes, regulate downstream biological processes, and thus are candidates for regulators of such tolerance of pearl millet. PgWRKY74 encodes a group IIc WRKY TF in pearl millet and is downregulated by drought. PgWRKY74 may have a role in drought tolerance. The objective of this study was to gain insights into the physiological and biochemical functions of PgWRKY74. Yeast one-hybrid and gel shift assays were performed to examine transcriptional activation potential and deoxyribonucleic acid (DNA)-binding ability, respectively. Transgenic Arabidopsis thaliana plants overexpressing PgWRKY74-green fluorescent protein (GFP) fusion gene were generated and tested for growth and stress-responsive gene expression under mannitol and NaCl-stressed conditions. A construct with PgWRKY74 enabled yeast reporter cells to survive on test media in the yeast one-hybrid assays. The electrophoretic mobility of DNA with putative WRKY TF-binding motifs was lower in the presence of a recombinant PgWRKY74 protein than its absence. The PgWRKY74-GFP-overexpressing Arabidopsis plants exhibited smaller rosette areas than did wild-type plants under mannitol-stressed and NaCl-stressed conditions, and exhibited weaker expression of RD29B, which is induced by the stress-related phytohormone abscisic acid (ABA), under the mannitol-stressed condition. PgWRKY74 have transcriptional activation potential and DNA-binding ability, and can negatively regulate plant responses to mannitol and NaCl stresses, possibly by decreasing ABA levels or ABA sensitivity.

Hydrolytic endonucleolytic ribozyme (HYER) is programmable for sequence-specific DNA cleavage.

Ribozymes are catalytic RNAs with diverse functions including self-splicing and polymerization. This work aims to discover natural ribozymes that behave as hydrolytic and sequence-specific DNA endonucleases, which could be repurposed as DNA manipulation tools. Focused on bacterial group II-C introns, we found that many systems without intron-encoded protein propagate multiple copies in their resident genomes. These introns, named HYdrolytic Endonucleolytic Ribozymes (HYERs), cleaved RNA, single-stranded DNA, bubbled double-stranded DNA (dsDNA), and plasmids in vitro. HYER1 generated dsDNA breaks in the mammalian genome. Cryo-electron microscopy analysis revealed a homodimer structure for HYER1, where each monomer contains a Mg2+-dependent hydrolysis pocket and captures DNA complementary to the target recognition site (TRS). Rational designs including TRS extension, recruiting sequence insertion, and heterodimerization yielded engineered HYERs showing improved specificity and flexibility for DNA manipulation.

Influence of Diabetes Mellitus and Universal Adhesive Application Mode on the Bond Strength of Composite Resin to Dentine.

The aim of the current study was to evaluate the influence of diabetes mellitus and the mode of applying a universal adhesive on the shear bond strength of composite resin to dentin. Forty teeth were extracted from diabetic individuals who had been living with diabetes for a period of 5 to 15 years. These forty teeth were divided into two groups: twenty molars from patients with type 1 diabetes and twenty molars from patients with type 2 diabetes. The remaining twenty sound human molars were collected from non-diabetic patients. The collected teeth were assigned into 3 groups, and each group was further divided into 2 subgroups. Group A (n = 20): non-diabetic (ND) where sub group IA: Universal adhesive applied in the total-etch mode (n = 10) and sub group IIA: Universal adhesive applied in the self-etch mode (n = 10). Group B (n = 20): diabetic type 1 (D1) where sub group IB: Universal adhesive applied in the total-etch mode (n = 10) and sub group IIB: Universal adhesive applied in the self-etch mode (n = 10). Group C (n = 20): diabetic type 2 (D2). Where Sub group IC: Universal adhesive applied in the total-etch mode (n = 10) and sub group IIC: Universal adhesive applied in the self-etch mode (n = 10). A Teflon mold measuring 3mm in diameter was attached to the dentin surface, used to build Filtek Z550 to a height of 3mm. The specimens were fixed to the universal testing to measure shear bond strength. There was a statistically significant difference of Mean ±SD of shear bond strength among the three tested groups. In group A, the Mean ±SD were (21.710 ± 0.638), it was decreased in group B to (14.626 ± 0.726) and group C to (17.740 ± 0.668). Subgroup I had lower shear bond strength values than subgroup II in all tested groups. The difference between each subgroup in groups A, B, and C was significant. 1. Diabetes mellitus has an adverse effect on the shear bond strength of composite to dentine. 2. Type 1 diabetes mellitus significantly reduces the shear bond strength of composite resin to dentin. 3. Shear bond strength of the universal adhesive was higher when applied to dentin using the self-etch mode, as compared to the total etch mode, in all groups, regardless of whether the participants had diabetes or not. Key words:Diabetes Mellitus, Universal Adhesive, Application Mode, Shear bond Strength, Dentine, Composite.

Correlation of mandibular third molar orientation and available retromolar space with arch length discrepancy in subjects with different growth pattern.

To evaluate, compare, and correlate the mandibular third molar orientation and available retromolar space with arch length discrepancy in subjects with skeletal class II malocclusion and different growth pattern. A total of 250 patients (age >18 yrs) having skeletal class II malocclusion (based on YEN angle and WITS appraisal) were divided into two groups. Both the groups (Group I with erupted mandibular third molars {N = 150} and Group II with impacted mandibular third molars {N = 100}) were subdivided into subgroups IA (n = 71), IB (n = 19), IC (n = 71) and Group IIA (n = 54), IIB (n = 30) and IIC (n = 16) for normo-, hypo- and hyperdivergent growth patterns, respectively (based on Jarabak ratio and Sn-Go-Gn angle). Four parameters, that is, retromolar space, width of third molar, third molar angulation, and mandibular incisor angulation were measured on orthopantomogram whereas arch length discrepancy was calculated with the help of lateral cephalogram and study model. Intragroup, intergroup comparisons (using unpaired Student's 't' test), and Pearson's correlation coefficient for assessed parameters were obtained. Third molar angulation and retromolar space were significantly higher in Group I than in Group II (hyperdivergent pattern of Group II had highest value). The width of third molar was less than retromolar space in Group II and vice versa for Group I. Mandibular incisor angulation and arch length discrepancy were more in Group II than in Group I, but difference was statistically nonsignificant. Strong positive correlation was observed for mandibular third molar angulation and available retromolar space in normo- and hyperdivergent growth patterns. Lack of retromolar space along with increased amount of arch length discrepancy and mandibular incisor angulation is responsible for increased chances of third molar impaction in some subjects with class II malocclusion.

Utility of combined use of elastography and von Willebrand factor antigen/thrombocyte ratio in prediction of gastroduodenal mucosal portal hypertension related vascular lesions

Background: Liver cirrhosis is the most important cause of portal hypertension, esophageal varices (EVs) represent its major complication as they may rupture and bleed with increased mortality rate. vWF-Ag is an established and valuable marker for determining the grade of fibrosis and cirrhosis and for mortality in patients with cirrhosis. Aim of our work is to search for the utility of combined use of elastography and Von Willebrand Factor Antigen/Thrombocyte Ratio in prediction of gastroduodenal portal hypertension related vascular lesions and identify suitable cutoff values for predicting these lesions. Patients and methods: 80 cirrhotic patients, were divided into 2 main groups, Group I included 20 patients with no evidence of portal hypertension related vascular lesions as verified by endoscopic examination (disease control group), group II, included patients with vascular lesions and was divided into 3 subgroups groups, group IIa included 20 patients with esophageal varices or gastro-esophageal varices, group IIb included 20 patients with isolated gastric varices, group IIc included 20 patients with portal hypertension related lesions other than esophageal and gastric varices.

In Silico Discovery of Group II Intron RNA Splicing Inhibitors.

Here, we describe the discovery of compounds that inhibit self-splicing in group II introns. Using docking calculations, we targeted the catalytic active site within the Oceanobacillus iheyensis group IIC intron and virtually screened a library of lead-like compounds. From this initial virtual screen, we identified three unique scaffolds that inhibit splicing in vitro. Additional tests revealed that an analog of the lead scaffold inhibits splicing in an intron-dependent manner. Furthermore, this analog exhibited activity against the group II intron from a different class: the yeast ai5γ IIB intron. The splicing inhibitors we identified could serve as chemical tools for developing group II intron-targeted antifungals, and, more broadly, our results highlight the potential of in silico techniques for identifying bioactive hits against structured and functionally complex RNAs.

Differential involvement of WRKY genes in abiotic stress tolerance of Dendrobium huoshanense

Extreme environments have adversely affected the habitats of medicinal Dendrobium species. Dendrobium huoshanense, a well-known traditional Chinese medicinal plant, is highly susceptible to abiotic stress. Evidence suggests that WRKY transcription factors play versatile roles in responding to both biotic and abiotic stressors experienced by terrestrial plants. The regulatory mechanisms underlying the stress responses of WRKY genes have been characterized for several model plants. In this study, 64 WRKY genes were identified in the D. huoshanense genome. Chromosome localization analysis showed that all the WRKY genes were anchored on the 15 pseudochromosomes, among which chromosome 5 was occupied by the highest number of WRKY genes (from DhWRKY7 to DhWRKY19), followed by chromosome 17 (from DhWRKY45 to DhWRKY53). Subcellular localization predictions indicated that most of the WRKY genes were located in the nucleus, with only a few WRKY genes localized in peroxisome, plastid, and mitochondria. Analysis of the conserved motifs and gene structure demonstrated that most genes, except for a few truncated genes, had more than two exons, and this was likely to be due to transposon insertion. A comparative genomics investigation based on the number of synteny blocks indicated the close evolutionary relationships of several orchid species, such as D. nobile, D. chrysotoxum, and G. elata, with D. huoshanense. Interspecies gene duplication events suggested that Dendrobium experienced a large-scale duplication event that included whole genome duplication and segmental duplication, and a limited number of WRKY paralogs were produced by tandem duplication. Based on the construction of a phylogenetic tree with AtWRKY nomenclature, DhWRKY genes could be categorized into three primary subfamilies. Group II could be further subdivided into five subclades. Group IIc had the highest number of WRKY genes, followed by Groups I and III. Multiple alignment of the amino acid sequences indicated that all DhWRKYs encoded a zinc-finger domain in addition to the highly conserved WRKYGQK domain, indicating their potential roles in transcriptional regulation and protein–protein interactions. Expression of most of the WRKY genes could be induced by light signals, phytohormones, and diverse abiotic stresses. Quantitative RT-PCR (qRT-PCR) and expression profiling results confirmed that the expression of WRKY genes could be induced by abscisic acid, salicylic acid, methyl jasmonate, gibberellin, and cold stress. Overall, the results of the cis-acting element analysis, expression profiling, and qRT-PCR highlighted DhWRKY42 as a key regulator involved in the regulation of jasmonic-acid-responsive genes. This study can provide valuable references for the study of WRKY genes in other Dendrobium species.

Light and scanning electron microscopic study of the effects of platelet-rich plasma on dexamethasone-induced osteoporosis in adult male albino rats

Abstract Background Osteoporosis may occur by using glucocorticoids for a long time. Recently, platelet-rich plasma (PRP) is used in treating a wide variety of musculoskeletal problems. Aim This work aimed to study the effect of PRP against dexamethasone-induced osteoporosis in adult male albino rats. Materials and methods This study was carried out on 40 adult male albino rats that were divided into two main groups: control group I: included 10 rats and experimental group II included 30 rats, which were subdivided into three subgroups. Subgroup IIa received PRP, subgroup IIb was injected with dexamethasone intramuscularly, and subgroup IIc received the same dose of dexamethasone and then injected with PRP. At the end of the experiment, upper ends of femur bones of all animals were obtained and processed for histological study using light and scanning electron micrograph studies. Results Light microscopic examination revealed osteoporotic histological changes in group IIb where cancellous bone trabeculae appeared thin and discontinuous. Scanning electron micrograph studies confirmed the results of the light microscopic study. Group IIc revealed partial preservation of the normal histological picture. Statistical analysis showed that group IIb and untreated left femur of group IIc revealed a highly significant decrease in the mean thickness of trabeculae and a highly significant increase in the width of intertrabecular spaces as compared with the control group, while group IIa and treated right femur of group IIc revealed nonsignificant changes as compared with the control group. Conclusion Osteoporosis resulted from glucocorticoids in adult male albino rats could be partially treated with PRP injection.

To evaluate and compare surface characteristics of different aesthetic nickel-titanium archwires using surface profilometry, sem and stereomicroscopy: A multi-arm split-mouth randomized controlled trial

This multi-arm split-mouth randomized controlled trial aimed to compare the surface characteristics, coating thickness and coating stability of four different aesthetic coated Nickel-Titanium Orthodontic rectangular archwires before and after oral exposure. To evaluate and compare the surface characteristics of four as-received (Control group) aesthetic nickel-titanium orthodontic rectangular archwires (0.016 x 0.022ʺ NiTi) from different manufacturers that are BioForce (Group I), Rabbit force (Group II), Libral (Group III), and ORMCO (Group IV) and after oral exposure (Experimental group) of four weeks, 15 patients meeting the inclusion criteria were recruited for this prospective study. Scanning electron microscopy, stereomicroscopy, and 3D profilometry were done for evaluating the surface characteristics of each group of wires.Four quadrants of every patient were randomized in a 1:1:1:1 ratio to the wires of either BioForce (Group I), Rabbit force (Group II), Libral (Group III), or ORMCO (Group IV). On examination of as-received wires (control group), the maximum coating thickness was seen in Group III c (Teflon coated, Libral), followed by Group IIc (Epoxy coated, Rabbit Force), then Group Ic (Teflon coated, BioForce), and least in Group IVc (Epoxy coated, ORMCO). After 4 weeks of oral exposure, the maximum coating loss was seen in Group II (Rabbit Force) wire, and the least amount of coating loss was seen in Group IV (ORMCO).The results of this study indicated a low aesthetic value of each of the manufacturer’s coated archwires because they presented a non-durable coating after oral exposure.

Apple WRKY transcription factor MdWRKY56 positively modulates drought stress tolerance

Drought seriously threatens the growth, development, and yield of crops. WRKY transcription factors (TFs) play important roles in abiotic stress responses. However, the study of WRKY TFs in woody plant apple (Malus domestica) in terms of drought tolerance regulation is limited. This study aimed to functionally characterize an apple group IIc WRKY gene MdWRKY56. The transcription abundance of MdWRKY56 was significantly upregulated under drought stress. The subcellular localization analysis revealed that the MdWRKY56 protein localized to the nucleus. Transcriptional activation experiments proved that MdWRKY56 had transcriptional activity in yeast, and its N-terminal played a decisive role in this activity. The functional analysis using MdWRKY56 transgenic Arabidopsis, apple calli, and apple plants indicated that the MdWRKY56 TF functioned positively in the modulation of osmotic and drought stress tolerance, with the MdWRKY56-overexpressed transgenic materials showing lower electrolyte leakage, malondialdehyde (MDA) content, and reactive oxygen species (ROS) accumulation, and higher fresh weight, proline content, and antioxidant enzyme activities. In conclusion, our findings indicated that MdWRKY56 served as an important positive regulator of drought stress tolerance in apple.

Molecular Cloning and Characterization of WRKY12, A Pathogen Induced WRKY Transcription Factor from Akebia trifoliata.

WRKY transcription factors (TFs), which are plant-specific TFs, play significant roles in plant defense. Here, a pathogen-induced WRKY gene, named AktWRKY12, which was the homologous gene of AtWRKY12, was isolated from Akebia trifoliata. The AktWRKY12 gene has a total length of 645 nucleotides and an open reading frame (ORF) encoding 214 amino acid polypeptides. The characterizations of AktWRKY12 were subsequently performed with the ExPASy online tool Compute pI/Mw, PSIPRED and SWISS-MODEL softwares. The AktWRKY12 could be classified as a member of WRKY group II-c TFs based on sequence alignment and phylogenetic analysis. The results of tissue-specific expression analysis revealed that the AktWRKY12 gene was expressed in all the tested tissues, and the highest expression level was detected in A. trifoliata leaves. Subcellular localization analysis showed that AktWRKY12 was a nuclear protein. Results showed that the expression level of AktWRKY12 significantly increased in A. trifoliata leaves with pathogen infection. Furthermore, heterologous over-expression of AktWRKY12 in tobacco resulted in suppressed expression of lignin synthesis key enzyme genes. Based on our results, we speculate that AktWRKY12 might play a negative role in A. trifoliata responding to biotic stress by regulating the expression of lignin synthesis key enzyme genes during pathogen infection.

Overexpression of a WRKY transcription factor McWRKY57-like from Mentha canadensis L. enhances drought tolerance in transgenic Arabidopsis

BackgroundDrought has become a major environmental problem affecting crop production. Members of the WRKY family play important roles in plant development and stress responses. However, their roles in mint have been barely explored.ResultsIn this study, we isolated a drought-inducible gene McWRKY57-like from mint and investigated its function. The gene encodes a group IIc WRKY transcription factor, McWRKY57-like, which is a nuclear protein with a highly conserved WRKY domain and a C2H2 zinc-finger structure, and has transcription factor activity. Its expression levels were examined in different tissues of mint and under the treatment of mannitol, NaCl, abscisic acid, and methyl jasmonate. We found that McWRKY57-like overexpression in Arabidopsis significantly increased drought tolerance. Further studies showed that under drought stress, McWRKY57-like-overexpressing plants had higher chlorophyll, soluble sugar, soluble protein, and proline contents but lower water loss rate and malondialdehyde content than wild-type plants. Moreover, the activities of antioxidant enzymes catalase, superoxide dismutase, and peroxidase were enhanced in McWRKY57-like transgenic plants. Furthermore, qRT-PCR analysis revealed that the drought-related genes AtRD29A, AtRD29B, AtRD20, AtRAB18, AtCOR15A, AtCOR15B, AtKIN2, and AtDREB1A were upregulated in McWRKY57-like transgenic plants than in wild-type Arabidopsis under simulated drought conditions.ConclusionThese data demonstrated that McWRKY57-like conferred drought tolerance in transgenic Arabidopsis by regulating plant growth, osmolyte accumulation and antioxidant enzyme activities, and the expression of stress-related genes. The study indicates that McWRKY57-like plays a positive role in drought response in plants.

Recognition of transcription terminators during retrotransposition: How to keep a group II intron quiet

Chung etal. recently presented the structure of a primitive group IIC intron with its DNA target, which reveals the structural requirements that this class of intron uses to recognize a transcription terminator stem loop at the DNA level for insertion during retrotransposition.