Search Results (16)

The germination of seeds is a prerequisite for crop production. Protrusion is important for seed germination, and visible radicle protrusion through seed covering layers is the second phase of the process of seed germination. Analyzing the mechanism of protrusion is important for the cultivation of rice varieties. In this study, 302 microcore germplasm populations were used for the GWAS of the protrusion percentage (PP). The frequency distribution of the PP at 48 h and 72 h is continuous, and six PP-associated QTLs were identified, but only qPP2 was detected repeatedly two times. The candidate gene analysis showed that LOC_Os02g57530 (ETR3), LOC_Os01g57610 (GH3.1) and LOC_Os04g0425 (CTB2) were the candidate genes for qPP2, qPP1 and qPP4, respectively. The haplotype (Hap) analysis revealed that Hap1 of ETR3, Hap1 and 3 of GH3.1 and Hap2 and 5 of CTB2 are elite alleles for the PP. Further validation of the germination phenotype of these candidate genes showed that Hap1 of ETR3 is a favorable allele for the germination percentage; Hap3 of GH3.1 is an elite allele for seed germination; and Hap5 of CTB2 is an elite allele for the PP, the germination percentage and the vigor index. The results of this study identified three putative candidate genes that provide valuable information for understanding the genetic control of seed protrusion in rice. Full article

Increasing the germination percentages from chemical and/or mechanical processes in native species is vital for the conservation of the species. The objective was to evaluate the effect of different treatments on the germination process of species belonging to the tropical deciduous forest (TDF) in a forest nursery. Eight species of trees and shrubs reported to be native to the TDF were selected; the seeds were collected from various sites of the state of Queretaro, Mexico. A randomized design of four replicates with 25 seeds was used, for which three pre-germinative scarification treatments were evaluated for each species: (1) thermal scarification (TS), (2) mechanical scarification (MS), and (3) chemical scarification (CS). Germination was considered as the protrusion of cotyledons onto the substrate and was recorded every 48 h for 60 days. A significant difference was observed between treatments (p ≤ 0.05). The mechanical treatment (MS) obtained the highest germination percentage and the least variation, in contrast to the thermal, chemical, and control treatments, which presented the lowest amount of germinated seeds. The MS treatment was the most effective for all the species studied because it allowed for 100% of the germination to be achieved. Full article

Caper is a perennial shrub that is widespread in the Mediterranean Basin. Although the fruits contain many seeds, they germinate slowly and with very low percentages, due to their nondeep physiological dormancy. The influence of the testa and endosperm, as well as the effect of applying gibberellic acid (GA₃) solutions on seed germination to release its dormancy, are reported in this study. The mechanical resistance exerted by the testa and endosperm against radicle protrusion in mature caper seeds was measured. The best germination results were obtained with seeds devoid of testa wetted with water and with intact seeds wetted with a 500 mg L⁻¹ GA₃ solution, without statistical differences between them. The GA₃ addition triggers an increase in both the content of endogenous gibberellins (GA) and the GA/abscisic acid ratio, increasing germination. Its germination consists of two temporally separated events: testa cracking and endosperm piercing. Testa cracking begins in the hilum-micropillar area; it involves a signal from the embryo, which GA can replace, possibly by increasing the growth potential of the embryo. After testa cracking, the radicle emerges through a hole in the micropylar endosperm. The puncture force necessary to pierce the micropylar endosperm decreased drastically during the first day of imbibition, remaining practically constant until testa cracking, decreasing afterwards, regardless of the addition or not of gibberellins. Full article

Cup overhanging in total hip arthroplasty is a predisposing factor to iliopsoas impingement. In dysplastic hips, cup implantation was simulated in an anatomic hip center of rotation (AHCR) and in high hip center (HHCR). We sought to assess: (1) the percentage of prominent cups; (2) quantify the cup protrusion at different sites on frontal, axial and sagittal views. In 40 Crowe III-IV hips, using a 3D CT-based planning software, cup planning in AHCR and HHCR (CR height ≥ 20 mm) was performed for every hip. Cup prominence was assessed on every plane. HHCR cups were less anteverted (p < 0.01), less medialized (p < 0.001) and less caudal (p = 0.01) than AHCR sockets. AHCR cups were more frequently prominent on at least one plane (92.5% vs. 77.5%), with minimal agreement between the two configurations (k = 0.31, p = 0.07). AHCR cups protruded more than HHCR sockets in the sagittal (p = 0.02) and axial planes (p < 0.001). Axially, at the center of the cup, prominence 6–11 mm occurred in nine (22.5%) AHCR and one (2.5%) HHCR socket. In conclusion, while a routine high hip center should not be recommended, cup placement at a center of rotation height < 20 mm is associated with higher rates and magnitudes of anterior cup protrusion in severe dysplasia. Full article

This work was conducted on the use of the RE (radicle emergence) test to estimate the seedling emergence (SE) and quality (seedling shoot weight (SSW), seedling root weight (SRW) and seedling height (SH)) of twelve hybrid cucumber seed lots with high germination (>95%) at low temperature conditions, high salt conditions and a combination of the two. The percentages of RE, which is a 2 mm radicle protrusion at 25 °C in the dark, were counted between 18 h and 32 h. The seeds were sown in peat moss in trays and kept at 15 °C (low temperature stress), or salty water was applied as 100 mM NaCl at 25 °C (salt stress) for three days, and both were applied in a combined-stress environment. Seedling emergence and quality parameters were reduced by both stress environments and their combination. SE, SSW, SRW and SH values ranged between: 91–100%, 614–844 mg, 102–143 mg, 6.8–8.8 cm at low temperature; 90–98%, 598–904 mg, 101–154 mg, 6.5–7.8 cm at salt stress; and 76–92%, 464–608 mg, 97–133 mg, 5.8–6.9 cm at their combination. The RE values with great differences seen among the lots (20–28 h) were regressed with seedling emergence and quality values. RE 24 h had the highest R² and significant values in all stress environments as R² = 0.596–0.858, p < 0.05–0.001 at low temperature; R² = 0.620–0.827, p < 0.05–0.001 with salt; and R² = 0.686–0.842, p < 0.05–0.001 with combined stresses. We concluded that RE as a vigour test can be used successfully to estimate the seedling quality of highly germinating hybrid cucumber seed lots. Use of the RE test in hybrid cucumber seeds and its influence on high-quality seedling production were discussed. Full article

In the present research work, PVFTX-100, PVFSDS, and PVFT-80 sponges were prepared using polyvinyl-alcohol (PVA) with surfactants triton X-100/sodium dodecyl sulfate (SDS)/Tween 80, respectively, for the removal of organic solvents from polluted soil/water samples. All three obtained sponges were further made hydrophobic using dodecyltrimethoxysilane (DTMS). The prepared sponges were characterized using different spectroscopic techniques and SEM analysis. The peaks obtained near 1050 cm⁻¹ and 790 cm⁻¹ were attributed to Si-O-C and alkyl side chain C-H stretching vibration that confirmed the formation of desired sponges. The SEM images showed the random roughness with a number of protrusions on sponge surfaces, which further played an important role in the absorption and retention of organic solvents molecules. The Sears method was chosen to calculate the surface area and pore volume of all the synthesized sponge samples. Among all three prepared sponges, the PVFTX-100 sponge showed a high pore volume and large surface area, with a maximum percentage absorption capacity of 96%, 91%, 89.9%, 85.6%, and 80 for chlorobenzene, toluene, diesel, petrol, and hexane, respectively, after eightcycles. The organic solvent uptake using PVFTX-100, PVFSDS, and PVFT-80 sponges is quite a unique and simple technology, which could be employed at a large scale for contaminated soil/water systems. Full article

Background and Objectives: One form of treatment for degenerative temporomandibular joint diseases such as osteoarthritis, rheumatic arthritis, TMJ ankylosis, and condylar resorption is total joint replacement. The aim of this study was to examine the function of the temporomandibular joint after prosthetic joint replacement. Materials and methods: Fifteen patients with unilateral or bilateral TMJ total joint replacements and 15 healthy controls were evaluated via a SICAT JMT+ device. This non-invasive system measures 3D position and linear movements in all degrees of freedom and allows undisturbed functional mandibular movements to provide a quantitative evaluation. In addition, a TMJ questionnaire consisting of the subjective symptoms was also obtained. To date, no similar studies have been cited in the literature. Results: Mandibular movements after prosthetic joint replacement were recorded during opening, closing, protrusion, and lateral excursive movements and were all significantly decreased compared to those of controls. In the treatment group, the maximum incisal opening was 33.46 ± 5.47 mm, left lateral movement was 1.91 ± 2.7 mm, right lateral movement was 1.74 ± 1.74 mm, and protrusive movement was 2.83 ± 2.05 mm. The p-value comparison study and control group indicated significant difference (p < 0.0001) between the two groups. The study group stated a high level of satisfaction with the total joint replacement. Conclusion: Within the limitations of the study, the following conclusions can be drawn: (1) TMJ replacement patients showed significantly limited jaw movements compared to the control group; (2) a small percentage of TMJ replacement patients still present low levels of pain but improved chewing ability and quality of life. Full article

The number, morphology, and distribution of C thyrocytes within the thyroid gland vary among species; however, studies in domestic animals are limited. In this study we compared the morphology, distribution pattern, and percentage of C thyrocytes in four domestic species: dogs, pigs, horses, and cattle. Eighty thyroid glands, 20 per species, were examined. C thyrocytes were visualized immunohistochemically with anti-calcitonin rabbit polyclonal antibody alone and combined with the periodic acid Schiff method to simultaneously visualize C thyrocytes with the basement membranes of thyroid follicles. C thyrocyte morphology varied considerably between species, from oval- (dogs) and spindle-shaped (pigs) to polymorphic (cattle and horses). Bovine C thyrocytes demonstrated cytoplasmic protrusion. C thyrocytes were located intrafolliculary (all species), epifollicularly (dogs, horses, cattle), or interfolicularly (cattle). Most porcine and bovine C thyrocytes existed individually whereas canine C thyrocytes usually formed clusters. In horses, they tended to form groups of various shapes and sizes or even rims encompassing whole follicles. In all species, the number of C thyrocyte profiles increased from the periphery to the central area of the thyroid lobe. The mean total fraction of C thyrocytes in the superficial, intermediate, and central areas were as follows: 2.55%, 8.43%, and 12.48% in dogs; 3.81%, 7.66%, and 10.79% in pigs; 1.55%, 7.44%, and 8.87% in horses; and 2.62%, 10.75%, and 12.96% in cattle. No statistical differences in the total number of C thyrocyte profiles were observed among species (8.87% in dogs, 8.58% in cattle, 7.98% in pigs, and 5.83% in horses). Our results indicated that the studied species displayed their own morphological characteristics and distribution pattern of C thyrocytes; however, total numbers of C thyrocyte profiles and their localization within the thyroid lobe are comparable. Full article

This paper focuses on the fabrication of high-quality novel products using a µEDM process variant called Reverse-µEDM. The tool plate required for the Reverse-µEDM is fabricated using Nd: YAG-based laser beam micromachining (LBµM) at the optimized process parameters. The Grey relation analysis technique is used for optimizing LBµM parameters for producing tool plates with arrayed micro-holes in elliptical and droplet profiles. Titanium sheets of 0.5 mm thickness were used for such micro-holes, which can be used as a Reverse-µEDM tool. The duty cycle (a combination of pulse width and frequency) and current percentage are considered as significant input process parameters for the LBµM affecting the quality of the micro-holes. A duty cycle of 1.25% and a current of 20% were found to be an optimal setting for the fabrication of burr-free shallow striation micro-holes with a minimal dimensional error. Thereafter, analogous protrusions of high dimensional accuracy and minimum deterioration were produced by Reverse-µEDM using the LBµM fabricated tool plates. Full article

We demonstrate a novel way of creating three-dimensional microfluidic channels capable of following complex topographies. To this end, substrates with open channels and different geometries were 3D-printed, and the open channels were consecutively closed with a thermoplastic using a low-resolution vacuum-forming approach. This process allows the sealing of channels that are located on the surface of complex multiplanar topographies, as the thermoplastic aligns with the surface-shape (the macrostructure) of the substrate, while the microchannels remain mostly free of thermoplastic as their small channel size resists thermoplastic inflow. This new process was analyzed for its capability to consistently close different substrate geometries, which showed reliable sealing of angles >90°. Furthermore, the thermoplastic intrusion into channels of different widths was quantified, showing a linear effect of channel width and percentage of thermoplastic intrusion; ranging from 43.76% for large channels with 2 mm width to only 5.33% for channels with 500 µm channel width. The challenging sealing of substrate ‘valleys’, which are created when two large protrusions are adjacent to each other, was investigated and the correlation between protrusion distance and height is shown. Lastly, we present three application examples: a serpentine mixer with channels spun around a cuboid, increasing the usable surface area; a cuvette-inspired flow cell for a 2-MXP biosensor based on molecular imprinted polymers, fitting inside a standard UV/Vis-Spectrophotometer; and an adapter system that can be manufactured by one-sided injection molding and is self-sealed before usage. These examples demonstrate how this novel technology can be used to easily adapt microfluidic circuits for application in biosensor platforms. Full article

Fossil frustules of Ellerbeckia and Melosira were studied using laboratory-based nano X-ray tomography (nano-XCT), transmission electron microscopy (TEM) and energy-dispersive X-ray spectroscopy (EDS). Three-dimensional (3D) morphology characterization using nondestructive nano-XCT reveals the continuous connection of fultoportulae, tube processes and protrusions. The study confirms that Ellerbeckia is different from Melosira. Both genera reveal heavily silicified frustules with valve faces linking together and forming cylindrical chains. For this cylindrical architecture of both genera, valve face thickness, mantle wall thickness and copulae thickness change with the cylindrical diameter. Furthermore, EDS reveals that these fossil frustules contain Si and O only, with no other elements in the percentage concentration range. Nanopores with a diameter of approximately 15 nm were detected inside the biosilica of both genera using TEM. In situ micromechanical experiments with uniaxial loading were carried out within the nano-XCT on these fossil frustules to determine the maximal loading force under compression and to describe the fracture behavior. The fracture force of both genera is correlated to the dimension of the fossil frustules. The results from in situ mechanical tests show that the crack initiation starts either at very thin features or at linking structures of the frustules. Full article

Background: Cilia are actin based cellular protrusions conserved from algae to complex multicellular organisms like Homo sapiens. Respiratory motile cilia line epithelial cells of the tracheobronchial tree, beat in a synchronous, metachronal wave, moving inhaled pollutants and pathogens cephalad. Their role in both congenital disorders like primary ciliary dyskinesia (PCD) to acquired disorders like chronic obstructive pulmonary disease (COPD) continues to evolve. In this current body of work we outline a protocol optimized to reciliate human nasal epithelial cells and mouse tracheal cells in vitro. Using this protocol, we knocked down known cilia genes, as well as use a small molecule inhibitor of Notch, N-[N-(3,5-Difluorophenacetyl)-L-alanyl]-S-phenylglycine t-butyl Ester (DAPT), to assess the effect of these on ciliogenesis in order to show the validity of our protocol. Methods: Tracheas were harvested from wild-type, adult C57B6 mice, pronase digested and sloughed off epithelial cells grown to confluence in stationary culture on rat-tail collagen coated wells. Upon reaching confluence, collagen was digested and cells placed suspension culture protocol to reciliate the cells. Using this suspension culture protocol, we employed siRNA gene knockdown to assay gene functions required for airway ciliogenesis. Knock down of Dynein axonemal heavy chain 5 (Dnah5), a ciliary structural protein, was confirmed using immunostaining. Mouse tracheal cells were treated in suspension with varying doses of DAPT, an inhibitor of Notch, with the purpose of evaluating its effect and dose response on ciliogenesis. The optimum dose was then used on reciliating human nasal epithelial cells. Results: siRNA knockdown of Foxj1 prevented ciliation, consistent with its role as a master regulator of motile cilia. Knockdown of Dnai1 and Dnah5 resulted in immotile cilia, and Cand1 knockdown, a centrosome protein known to regulate centrosome amplification, inhibited airway ciliogenesis. Dnah5 knockdown was confirmed with significantly decreased immunostaining of cilia for this protein. Inhibiting Notch signaling by inhibiting gamma secretase with DAPT enhanced the percentage of ciliation, and resulted in longer cilia that beat with higher frequency in both mouse and human airway epithelia. Conclusions: Modifying existing reciliation protocols to suit both human nasal epithelial and mouse tracheal tissue, we have shown that knockdown of known cilia-related genes have the expected effects. Additionally, we have demonstrated the optimal dosage for significantly improving reciliation of airway epithelia using DAPT. Given that cilia length and function are significantly compromised in COPD, these findings open up interesting avenues for further exploration. Full article

Pinus bungeana Zucc. ex Endl. is an endemic conifer tree species in China with high ornamental value. In order to investigate favorable conditions for seed germination and explore the germination inhibition mechanism of this species at high temperatures, the effects of temperature, light, and storage on the mean germination time (MGT), speed of germination (SG), and total germination percentage (TGP) are evaluated here. Seeds that have either been kept still or entered into a state of dormancy at high temperature are assessed here by a recovery experiment. Furthermore, the contribution of covering layers on thermo-inhibition is analyzed here, including the way they work. This has been realized by the structural observation and via the determination of the abscisic acid (ABA) content. The results show that seeds germinate to a high percentage (approximately 90%) at temperatures of 15 or 20 °C, with or without light, whereas higher temperatures of 25 or 30 °C impeded radicle protrusion and resulted in the germination percentage decreasing sharply (within 5%). Inhibition at high temperatures was thoroughly reversed (bringing about approximately 80% germination) by placing the ungerminated seeds in favorable temperatures and incubating them for an additional 30 days. Dry cold storage did little to reduce the temperature request for germination. Embryo coverings, especially the nucellar membrane, and ABA levels both had a dominant role in seed germination regulation in response to temperature. Under favorable temperature conditions, the levels of ABA significantly decreased. Germination occurred when the levels dropped to a threshold of 15 ng/g (FW (Fresh Weight)). Incubation at a high temperature (25 °C) greatly increased ABA levels and caused the inhibition of radicle protrusion. Full article

Triple-negative breast cancer (TNBC) is an aggressive form of breast cancer with high risk of relapse and metastasis. TNBC is a heterogeneous disease comprising different molecular subtypes including those with mesenchymal features. The tyrosine kinase AXL is expressed in mesenchymal cells and plays a role in drug resistance, migration and metastasis. We confirm that AXL is more expressed in mesenchymal TNBC cells compared to luminal breast cancer cells, and that its invalidation impairs cell migration while having no or little effect on cell viability. Here, we found that AXL controls directed migration. We observed that AXL displays a polarized localization at the Golgi apparatus and the leading edge of migratory mesenchymal TNBC cells. AXL co-localizes with F-actin at the front of the cells. In migratory polarized cells, the specific AXL inhibitor R428 displaces AXL and F-actin from the leading edge to a lateral area localized between the front and the rear of the cells where both are enriched in protrusions. In addition, R428 treatment disrupts the polarized localization of the Golgi apparatus towards the leading edge in migratory cells. Immunohistochemical analysis of aggressive chemo-resistant TNBC samples obtained before treatment reveals inter- and intra-tumor heterogeneity of the percentage of AXL expressing tumor cells, and a preference of these cells to be in contact with the stroma. Taken together, our study demonstrates that AXL controls directed cell migration most likely by regulating cell polarity. Full article

In order to metastasize, tumor cells need to migrate and invade the surrounding tissues. It is important to identify compound(s) capable of disrupting the metastasis of invasive cancer cells, especially for hindering invadopodia formation, so as to provide anti-metastasis targeted therapy. Invadopodia are thought to be specialized actin-rich protrusions formed by highly invasive cancer cells to degrade the extracellular matrix (ECM). A curcuminoid analogue known as 2,6-bis-(4-hydroxy-3-methoxybenzylidine)cyclohexanone or BHMC has shown good potential in inhibiting inflammation and hyperalgesia. It also possesses an anti-tumor effects on 4T1 murine breast cancer cells in vivo. However, there is still a lack of empirical evidence on how BHMC works in preventing human breast cancer invasion. In this study, we investigated the effect of BHMC on MDA-MB-231 breast cancer cells and its underlying mechanism of action to prevent breast cancer invasion, especially during the formation of invadopodia. All MDA-MB-231 cells, which were exposed to the non-cytotoxic concentrations of BHMC, expressed the proliferating cell nuclear antigen (PCNA), which indicate that the anti-proliferative effects of BHMC did not interfere in the subsequent experiments. By using a scratch migration assay, transwell migration and invasion assays, we determined that BHMC reduces the percentage of migration and invasion of MDA-MB-231 cells. The gelatin degradation assay showed that BHMC reduced the number of cells with invadopodia. Analysis of the proteins involved in the invasion showed that there is a significant reduction in the expressions of Rho guanine nucleotide exchange factor 7 (β-PIX), matrix metalloproteinase-9 (MMP-9), and membrane type 1 matrix metalloproteinase (MT1-MMP) in the presence of BHMC treatment at 12.5 µM. Therefore, it can be postulated that BHMC at 12.5 µM is the optimal concentration for preventing breast cancer invasion. Full article