Specific interaction of TFII-I with an upstream element on the HIV-1 LTR regulates induction of latent provirus

RBF-2 is a factor comprised of a USF1/2 heterodimer, whose association with a highly conserved upstream element (RBEIII) on the HIV-1 LTR requires a co-factor TFII-I. We have identified specific nucleotides, immediately 3′ of RBEIII that are required for stable association of TFII-I with this region of the LTR. Mutations that inhibit interaction of TFII-I with DNA also prevent stimulation of USF binding to RBEIII, and render the integrated LTR unresponsive to T cell signaling. These results demonstrate an essential role of TFII-I bound at an upstream LTR element for viral replication.     

miR-142-3p/5p role in cancer: From epigenetic regulation to immunomodulation

MicroRNAs (miRNAs) play critical roles in cancer pathobiology, acting as regulators of gene expression and pivotal drivers of tumorigenesis. It is believed that miRNAs act through canonical mechanisms, involving the binding of mature miRNAs to target messenger RNAs (mRNAs) and subsequent repression of protein translation or degradation of target mRNAs. miR-142-3p/5p has been extensively studied and established as a key regulator in various malignancies. Recent discoveries have revealed miR-142-3p/5p serve as either oncogene or tumor suppressor in cancer. By targeting epigenetic factor and cancer-related signaling pathway, miR-142-3p/5p can regulate wide range of downstream genes. The immune modulatory role of miR-142-3p/5p has been shown in various cancers, which provides significant insight into immunosuppression and tumor escape from the immune response. Exosomes with miR-142-3p/5p facilitate cell communication and can affect cancer cell behavior, offering potential therapeutic, and diagnosis applications in cancer therapy. In this review, for the first time, we comprehensively summarize the current knowledge regarding mentioned functions of miR-142-3p/5p in cancer pathobiology.     

Effect of Mechanical Forces on the Behavior of Dental Stem Cells: A Scoping Review of <i>In-Vitro</i> Studies

This article is a scoping review of the studies that assessed the effect of mechanical forces on the behavior of dental stem cells (DSCs). PubMed and Scopus searches were done for in-vitro studies evaluating the effect of tension, hydrostatic pressure (i.e., the pressure applied through an incompressible fluid), compression, simulated microgravity, and vibration on DSCs. The following factors were analyzed: osteogenic/odontogenic differentiation, proliferation, adhesion and migration. Articles were reviewed according to the Preferred Reporting Items for Systematic Reviews extension for scoping reviews (PRISMA-ScR) guideline. Included studies were evaluated based on the modified Consolidated Standards of Reporting Trials (CONSORT). A total of 18 studies published from 2008–2019 were included. Nine studies were focusing on Periodontal ligament Stem Cells (PDLSCs), eight studies on Dental Pulp Stem Cells (DPSCs) and one study on Stem Cells from Apical Papilla (SCAP). Results showed that tension, three-dimensional stress and simulated microgravity promoted the proliferation and osteogenic differentiation of PDLSCs. DPSCs proliferation increased after microgravity and tension exertion. In addition, dynamic hydrostatic pressure and compression promoted odontogenic differentiation of DPSCs. Besides, mechanical stimuli increased the osteogenic differentiation of DPSCs. One study analyzed the effect of carrier features on the response of DSCs to 3D-stress and showed that cells cultivated on scaffolds with 30% bioactive glass (BAG) had the highest osteogenic differentiation compared to other ratios of BAG. It has been shown that increasing the duration of tension (i.e., from 3 h to 24 h force application) enhanced the positive effect of force application on the osteogenic differentiation of DSCs. In conclusion, all types of mechanical forces except uniaxial tension increased the osteogenic/odontogenic differentiation of DSCs. In addition, the effect of mechanical stimulation on the proliferation of DSCs differs based on the type of stem cells and mechanical force.     

Interlaboratory reproducibility of selective reaction monitoring assays using multiple upfront analyte enrichment strategies

Over the past few years, mass spectrometry has emerged as a technology to complement and potentially replace standard immunoassays in routine clinical core laboratories. Application of mass spectrometry to protein and peptide measurement can provide advantages including high sensitivity, the ability to multiplex analytes, and high specificity at the amino acid sequence level. In our previous study, we demonstrated excellent reproducibility of mass spectrometry-selective reaction monitoring (MS-SRM) assays when applying standardized standard operating procedures (SOPs) to measure synthetic peptides in a complex sample, as lack of reproducibility has been a frequent criticism leveled at the use of mass spectrometers in the clinical laboratory compared to immunoassays. Furthermore, an important caveat of SRM-based assays for proteins is that many low-abundance analytes require some type of enrichment before detection with MS. This adds a level of complexity to the procedure and the potential for irreproducibility increases, especially across different laboratories with different operators. The purpose of this study was to test the interlaboratory reproducibility of SRM assays with various upfront enrichment strategies and different types of clinical samples (representing real-world body fluids commonly encountered in routine clinical laboratories). Three different, previously published enrichment strategies for low-abundance analytes and a no-enrichment strategy for high-abundance analytes were tested across four different laboratories using different liquid chromatography-SRM (LC-SRM) platforms and previously developed SOPs. The results demonstrated that these assays were indeed reproducible with coefficients of variation of less than 30% for the measurement of important clinical proteins across all four laboratories in real world samples.     

A knockout mutation in the lignin biosynthesis gene CCR1 explains a major QTL for acid detergent lignin content in Brassica napus seeds

Seed coat phenolic compounds represent important antinutritive fibre components that cause a considerable reduction in value of seed meals from oilseed rape (Brassica napus). The nutritionally most important fibre compound is acid detergent lignin (ADL), to which a significant contribution is made by phenylpropanoid-derived lignin precursors. In this study, we used bulked-segregant analysis in a population of recombinant inbred lines (RILs) from a cross of the Chinese oilseed rape lines GH06 (yellow seed, low ADL) and P174 (black seed, high ADL) to identify markers with tight linkage to a major quantitative trait locus (QTL) for seed ADL content. Fine mapping of the QTL was performed in a backcross population comprising 872 BC₁F₂ plants from a cross of an F₇ RIL from the above-mentioned population, which was heterozygous for this major QTL and P174. A 3:1 phenotypic segregation for seed ADL content indicated that a single, dominant, major locus causes a substantial reduction in ADL. This locus was successively narrowed to 0.75 cM using in silico markers derived from a homologous Brassica rapa sequence contig spanning the QTL. Subsequently, we located a B. rapa orthologue of the key lignin biosynthesis gene CINNAMOYL CO-A REDUCTASE 1 (CCR1) only 600 kbp (0.75 cM) upstream of the nearest linked marker. Sequencing of PCR amplicons, covering the full-length coding sequences of Bna.CCR1 homologues, revealed a locus in P174 whose sequence corresponds to the Brassica oleracea wild-type allele from chromosome C8. In GH06, however, this allele is replaced by a homologue derived from chromosome A9 that contains a loss-of-function frameshift mutation in exon 1. Genetic and physical map data infer that this loss-of-function allele has replaced a functional Bna.CCR1 locus on chromosome C8 in GH06 by homoeologous non-reciprocal translocation.     

Effect of temperature on development time and life table parameters of Nephus hiekei Fürsch,the important predator of Phenacoccus solenopsis Tinsley

The cotton mealybug, Phenacoccus solenopsis Tinsley (Hemiptera: Pseudococcidae) has become a serious threat to agricultural and ornamental plants in Southwest Iran. Pest-control studies have found Nephus hiekei Fürsch (Coleoptera: Coccinellidae), in large numbers on Phenacoccus solenopsis Tinsley, as a potential predator. However, there has been no investigation on this predator. In this research, the biology and age-stage, two-sex life table parameters of N. hiekei feeding on Ph. solenopsis were investigated at three constant temperatures (27, 32 and 37 ± 1 °C, 65 ± 5% RH and a photoperiod of 16: 8 (L: D) h. The duration of total pre-adult stage was found to decrease raising temperature from 26.98 ± 0.36 days at 27 °C to 14.24 ± 0.27 days at 37 °C. The oviposition period lasted 63.95 ± 3.14, 66.42 ± 3.34 and 25.20 ± 1.59 days at 27, 32, and 37 °C, respectively. Females laid an average of 365.68 ± 22.36, 543.79 ± 27.27 and 106.25 ± 6.38 eggs, at these three temperatures, respectively. The intrinsic rate of increase (r_m = 0.1333 ± 0.0050 d^–1), finite rate of increase (λ = 1.1423 ± 0.0057 d^–1) and net reproductive rate (R₀ = 188.89 ± 28.34 offspring) as well as gross reproductive rate (GRR = 308.31 ± 39.54 offspring) were greatest at 32 °C. The shortest mean generation time (T = 26.74 ± 0.65 days) was recorded at 37 °C and the longest at 27 °C (T = 56.48 ± 1.0 days). These results indicate that N. hiekei can successfully survive and reproduce at temperatures of around 32 °C, and has good potential to be an effective biological control agent of Ph. solenopsis.     

Thymoquinone Inhibits Murine Leukemia WEHI-3 Cells In Vivo and In Vitro

BackgroundThymoquinone is an active ingredient isolated from Nigella sativa (Black Seed). This study aimed to evaluate the in vitro and in vivo anti-leukemic effects of thymoquinone on WEHI-3 cells.ConclusionThymoquinone promoted natural killer cell activities. This compound showed high toxicity against WEHI-3 cell line which was confirmed by an increase of the early apoptosis, followed by up-regulation of the anti-apoptotic protein, Bcl2, and down-regulation of the apoptotic protein, Bax. On the other hand, high reduction of the spleen and liver weight, and significant histopathology study of spleen and liver confirmed that thymoquinone inhibited WEHI-3 growth in the BALB/c mice. Results from this study highlight the potential of thymoquinone to be developed as an anti-leukemic agent.     

Andrographis paniculata Leaf Extract Prevents Thioacetamide-Induced Liver Cirrhosis in Rats

This study investigated the hepatoprotective effects of ethanolic Andrographis paniculata leaf extract (ELAP) on thioacetamide-induced hepatotoxicity in rats. An acute toxicity study proved that ELAP is not toxic in rats. To examine the effects of ELAP in vivo, male Sprague Dawley rats were given intraperitoneal injections of vehicle 10% Tween-20, 5 mL/kg (normal control) or 200 mg/kg TAA thioacetamide (to induce liver cirrhosis) three times per week. Three additional groups were treated with thioacetamide plus daily oral silymarin (50 mg/kg) or ELAP (250 or 500 mg/kg). Liver injury was assessed using biochemical tests, macroscopic and microscopic tissue analysis, histopathology, and immunohistochemistry. In addition, HepG2 and WRL-68 cells were treated in vitro with ELAP fractions to test cytotoxicity. Rats treated with ELAP exhibited significantly lower liver/body weight ratios and smoother, more normal liver surfaces compared with the cirrhosis group. Histopathology using Hematoxylin and Eosin along with Masson’s Trichrome stain showed minimal disruption of hepatic cellular structure, minor fibrotic septa, a low degree of lymphocyte infiltration, and minimal collagen deposition after ELAP treatment. Immunohistochemistry indicated that ELAP induced down regulation of proliferating cell nuclear antigen. Also, hepatic antioxidant enzymes and oxidative stress parameters in ELAP-treated rats were comparable to silymarin-treated rats. ELAP administration reduced levels of altered serum liver biomarkers. ELAP fractions were non-cytotoxic to WRL-68 cells, but possessed anti-proliferative activity on HepG2 cells, which was confirmed by a significant elevation of lactate dehydrogenase, reactive oxygen species, cell membrane permeability, cytochrome c, and caspase-8,-9, and, -3/7 activity in HepG2 cells. A reduction of mitochondrial membrane potential was also detected in ELAP-treated HepG2 cells. The hepatoprotective effect of 500 mg/kg of ELAP is proposed to result from the reduction of thioacetamide-induced toxicity, normalizing reactive oxygen species levels, inhibiting cellular proliferation, and inducing apoptosis in HepG2 cells.