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This study systematically investigates the effects of the coaxial nozzle’s inner nozzle diameter on the strength and gelation of filaments produced via extrusion-based 3D printing with in situ ionic crosslinking. In this system, bioink (sodium alginate solution) was extruded through the outer nozzle, and the ionic crosslinking solution (calcium chloride solution) was extruded through the inner nozzle. The outer nozzle diameter was fixed at 2.16 mm, and the inner nozzle diameter was varied among 1.19, 0.84, and 0.584 mm. The results indicate that, as the inner nozzle diameter decreased, filament strength decreased, and filament gelation became poorer. These findings highlight the importance of optimizing inner nozzle diameter for improved filament strength and gelation in extrusion-based 3D printing with in situ ionic crosslinking.

This paper reports an experimental study on the compatibility of human bronchial epithelial (HBE) cells in a collagen–alginate bioink. The compatibility was assessed using the culture well method with three bioink compositions prepared from a 10% alginate solution and neutralized TeloCol-10 mg/mL collagen stock solution. Cell viability, quantified by (live cell count—dead cell count)/live cell count within the HBE cell-laden hydrogel, was evaluated using the live/dead assay method from Day 0 to Day 6. Experimental results demonstrated that the collagen–alginate 4:1 bioink composition exhibited the highest cell viability on Day 6 (85%), outperforming the collagen–alginate 1:4 bioink composition and the alginate bioink composition, which showed cell viability of 75% and 45%, respectively. Additionally, the live cell count was highest for the collagen–alginate 4:1 bioink composition on Day 0, a trend that persisted through Days 1 to 6, underscoring its superior performance in maintaining cell viability and promoting cell proliferation. These findings show that the compatibility of HBE cells with the collagen–alginate 4:1 bioink composition was higher compared with the other two bioink compositions.

Abstract Crapemyrtle Bark Scale (Acanthococcus lagerstroemiae; CMBS) is an invasive pest species that primarily infest crapemyrtles (Lagerstroemia spp.) in the United States. Recent reports have revealed the dire threat of CMBS to attack not only crapemrytles but also the U.S. native species with expanded host plants such as American beautyberry (Callicarpa spp.) and Hypericum kalmianum L. (St. Johnswort). A better understanding of plant–insect interaction will provide better and environmental-friendly pest management strategies. In this study, we constructed the first comprehensive life table for CMBS to characterize its biological parameters, including developmental stages, reproductive behavior, and fecundity. The indirect effects of three plant nutrient conditions (water, 0.01MS, and 0.1MS) on CMBS populations were examined using the age-stage, two-sex life table. The demographic analyses revealed that the plant nutrient conditions had significantly altered CMBS development in terms of the intrinsic rate of increase (r), the finite rate of increase (λ), the net reproductive rate (R 0 ), and mean generation time (T). Higher r, λ, and R 0 were recorded under nutrient-deficient conditions (water), while CMBS reared on plants with healthier growing conditions (0.1MS) had the most prolonged T. Overall, CMBS shows better insect performance when reared on plants under nutrient-deficient conditions.

Crapemyrtle bark scale [CMBS (Acanthococcus lagerstroemiae)], a newly emerged pest in the United States, has spread to 16 U.S. states and unexpectedly spread on a native species american beautyberry (Callicarpa americana) in Texas and Louisiana in 2016 since it was initially reported on crapemyrtles (Lagerstroemia sp.) in Texas in 2004. The infestation of CMBS negatively impacted the flowering of crapemyrtles. We observed the infestation on the two most commercially available edible fig (Ficus carica) cultivars Beer’s Black and Chicago Hardy in a preliminary trial in 2018. To help estimate CMBS potential in aggravating risks to the ecosystem stability and the green industry, we conducted a host range and suitability test using ‘Bok Tower’ american beautyberry as a positive control with other eight beautyberry (Callicarpa) species [mexican beautyberry (C. acuminata), ‘Profusion’ bodinieri beautyberry (C. bodinieri), ‘Issai’ purple beautyberry (C. dichotoma), japanese beautyberry (C. japonica var. luxurians), ‘Alba’ white-fruited asian beautyberry (C. longissima), taiwan beautyberry (C. pilosissima), luanta beautyberry (C. randaiensis), and willow-leaf beautyberry (C. salicifolia)] and three fig (Ficus) species [creeping fig (F. pumila), roxburgh fig (F. auriculata), and waipahu fig (F. tikoua)] over 25 weeks. All the tested beautyberry species and waipahu fig sustainably supported the development and reproduction of nymphal CMBS and were confirmed as CMBS hosts. Furthermore, comparing with the control, mexican beautyberry, ‘Profusion’ bodinieri beautyberry, taiwan beautyberry, and willow-leaf beautyberry were significantly less suitable, while ‘Issai’ purple beautyberry, japanese beautyberry, ‘Alba’ white-fruited asian beautyberry, and luanta beautyberry were as suitable as ‘Bok Tower’ american beautyberry. Thus, when using beautyberries in landscapes, their different potential to host CMBS should be considered to minimize spreading CMBS through the native ecosystems.

Crapemyrtle bark scale (CMBS; Acanthococcus lagerstroemiae Kuwana) is an invasive insect that was first discovered in the United States in 2004. The polyphagous feeding habit of CMBS has allowed it to infest a wide range of plant species beyond its primary host, Lagerstroemia. Using molecular approaches, we studied the genetic relationships between CMBS specimens and their hosts from different geographic locations. Naturally occurring CMBS infestations were confirmed on American beautyberry (Callicarpa americana L.), a native plant species in the United States, and spirea (Spiraea L.). The new infestation of CMBS found on Spiraea raises the alarm that other economically important crops in the Amygdaloideae subfamily (subfamily under Rosaceae) might be susceptible to CMBS attacks.

Summary: The sensory organelle cilium is involved in sensing and transducing important signaling cascades in almost all cells of our body. These ciliary-mediated pathways affect cellular homeostasis and metabolisms profoundly. However, it is almost completely unknown whether the cellular metabolic state affects the assembly of cilia. This study is to investigate how O-linked β-N-acetylglucosamine (O-GlcNAc), a sensor of cellular nutrients, regulates the cilia length. Pharmacologic or genetic inhibition of O-GlcNAcylation led to longer cilia, and vice versa. Further biochemical assays revealed that both α-tubulin and HDAC6 (histone deacetylase 6) were O-GlcNAcylated in vivo. In vitro enzymatic assays showed that O-GlcNAcylation of either tubulin or HDAC6 promoted microtubule disassembly, which likely in turn caused ciliary shortening. Taken together, these results uncovered a negative regulatory role of O-GlcNAc in modulating the ciliary microtubule assembly. The cross talk between O-GlcNAc and cilium is likely critical for fine-tuning the cellular response to nutrients. : Biological Sciences; Molecular Biology; Cell Biology; Functional Aspects of Cell Biology Subject Areas: Biological Sciences, Molecular Biology, Cell Biology, Functional Aspects of Cell Biology

Host range confirmation of invasive hemipterans relies on the evaluation of plant susceptibility though greenhouse or field trials, which are inefficient and time-consuming. When the green industry faces the fast-spreading threat of invasive pests such as crapemyrtle bark scale (Acanthococcus lagerstroemiae), it is imperative to timely identify potential host plants and evaluate plant resistance/susceptibility to pest infestation. In this study, we developed an alternative technology to complement the conventional host confirmation methods. We used electrical penetration graph (EPG) based technology to monitor the A. lagerstroemiae stylet-tip position when it was probing in different plant tissues in real-time. The frequency and relative amplitude of insect EPG waveforms were extracted by an R programming-based software written to generate eleven EPG parameters for comparative analysis between plant species. The results demonstrated that the occurrences of phloem phase and xylem phase offered conclusive evidence for host plant evaluation. Furthermore, parameters including the percentage of insects capable of accessing phloem tissue, time duration spent on initiating phloem phase and ingesting phloem sap, provided insight into why host plant susceptibility differs among similar plant species. In summary, this study developed a novel real-time diagnostic tool for quick A. lagerstroemiae host confirmation, which laid the essential foundation for effective pest management.

Process variables of bioprinting (including extrusion pressure, nozzle size, and bioink composition) can affect the shape fidelity and cell viability of printed constructs. Reported studies show that increasing extrusion pressure or decreasing nozzle size would decrease cell viability in printed constructs. However, a smaller nozzle size is often necessary for printing constructs of higher shape fidelity, and a higher extrusion pressure is usually needed to extrude bioink through nozzles with a smaller diameter. Because values of printing process variables that increase shape fidelity can be detrimental to cell viability, the optimum combination of variables regarding both shape fidelity and cell viability must be determined for specific bioink compositions. This paper reports a designed experimental investigation (full factorial design with three variables and two levels) on bioprinting by applying layer-by-layer photo-crosslinking and using the alginate-methylcellulose-GelMA bioink containing algae cells. The study investigates both the main effects and interaction effects of extrusion pressure, nozzle size, and bioink composition on the shape fidelity and cell viability of printed constructs. Results show that, as extrusion pressure changed from its low level to its high level, shape fidelity and cell viability decreased. As nozzle size changed from its low level to its high level, shape fidelity decreased while cell viability increased. As bioink composition changed from its low level (with more methylcellulose content in the bioink) to its high level (with less methylcellulose content in the bioink), shape fidelity and cell viability increased.

This review discusses the reported studies investigating the use of bioprinting to develop functional organ-on-chip systems from a manufacturing perspective. These organ-on-chip systems model the liver, kidney, heart, lung, gut, bone, vessel, and tumors to demonstrate the viability of bioprinted organ-on-chip systems for disease modeling and drug screening. In addition, the paper highlights the challenges involved in using bioprinting techniques for organ-on-chip system fabrications and suggests future research directions. Based on the reviewed studies, it is concluded that bioprinting can be applied for the automated and assembly-free fabrication of organ-on chip systems. These bioprinted organ-on-chip systems can help in the modeling of several different diseases and can thereby expedite drug discovery by providing an efficient platform for drug screening in the preclinical phase of drug development processes.

Crapemyrtle bark scale (CMBS, Acanthococcus lagerstroemiae), an invasive polyphagous sap-sucking hemipteran, has spread across 14 states of the United States since 2004. The infestation of CMBS has negatively impacted the flowering of ornamental plants and even the fruiting of some crops. Host identification is critical for determining potential risks in ecosystems and industries and helps develop strategic management. A host confirmation test was performed over 25 weeks using six Lagerstroemia species (L. caudata, L. fauriei ‘Kiowa’, L. indica ‘Dynamite’, L. limii, L. speciosa, and L. subcostata) and California loosestrife (Lythrum californicum). The 25-week observations confirmed all tested plants as the hosts. The repeated measures of analysis of variance (ANOVA; Tukey’s HSD, α = 0.05) indicated that the average number of CMBS females differed significantly between L. limii and L. speciosa. The highest number of the females observed on L. limii was 576 ± 25 (mean ± SE) at 17 weeks after inoculation (WAI), while the highest number was 57 ± 15 on L. speciosa at 19 WAI. In addition, L. subcostata and L. speciosa had significantly high and low numbers of males, respectively, among the Lagerstroemia species. Our results suggest that L. speciosa could be incorporated in developing new cultivars with low CMBS suitability.

Crapemyrtle bark scale (CMBS; Acanthococcus lagerstroemiae) is an exotic pest species that causes aesthetic and economic damage to crapemyrtles and poses potential threats to other horticultural crops in the United States. Although previous studies reported the infestation of CMBS on several alternative hosts across multiple families in Asia, its potential threats to other documented alternative hosts remain elusive and yet to be confirmed. In this study, feeding preference studies of CMBS were conducted on forty-nine plant species and cultivars in 2016 and 2019, in order to gain insight into the expansion of CMBS distribution in the United States, as well as other regions of the world. The infestations of CMBS were confirmed on apple (Malus domestica), Chaenomeles speciosa, Disopyros rhombifolia, Heimia salicifolia, Lagerstroemia ‘Spiced Plum’, M. angustifolia, and twelve out of thirty-five pomegranate cultivars. However, the levels of CMBS infestation on these test plant hosts in this study is very low compared to Lagerstroemia, and may not cause significant damage. No sign of CMBS infestation was observed on Rubus ‘Arapaho’, R. ‘Navaho’, R. idaeus ‘Dorman Red’, R. fruticosus, B. microphylla var. koreana × B. sempervirens, B. harlandii, or D. virginiana.

Although microtubules are known for dynamic instability, the dynamicity is considered to be tightly controlled to support a variety of cellular processes. Yet diverse evidence suggests that this is not applicable to Chlamydomonas, a biflagellate fresh water green alga, but intense autofluorescence from photosynthesis pigments has hindered the investigation. By expressing a bright fluorescent reporter protein at the endogenous level, we demonstrate in real time discreet sweeping changes in algal microtubules elicited by rises of intracellular H+ and Na+. These results from this model organism with characteristics of animal and plant cells provide novel explanations regarding how pH may drive cellular processes; how plants may respond to, and perhaps sense stresses; and how organisms with a similar sensitive cytoskeleton may be susceptible to environmental changes.

Cilia/flagella are assembled and maintained by the process of intraflagellar transport (IFT), a highly conserved mechanism involving more than 20 IFT proteins. However, the functions of individual IFT proteins are mostly unclear. To help address this issue, we focused on a putative IFT protein TTC26/DYF13. Using live imaging and biochemical approaches we show that TTC26/DYF13 is an IFT complex B protein in mammalian cells and Chlamydomonas reinhardtii. Knockdown of TTC26/DYF13 in zebrafish embryos or mutation of TTC26/DYF13 in C. reinhardtii, produced short cilia with abnormal motility. Surprisingly, IFT particle assembly and speed were normal in dyf13 mutant flagella, unlike in other IFT complex B mutants. Proteomic and biochemical analyses indicated a particular set of proteins involved in motility was specifically depleted in the dyf13 mutant. These results support the concept that different IFT proteins are responsible for different cargo subsets, providing a possible explanation for the complexity of the IFT machinery.

Intraflagellar transport (IFT), the key mechanism for ciliogenesis, involves large protein particles moving bi-directionally along the entire ciliary length. IFT particles contain two large protein complexes, A and B, which are constructed with proteins in a core and several peripheral proteins. Prior studies have shown that in Chlamydomonas reinhardtii, IFT46, IFT52, and IFT88 directly interact with each other and are in a subcomplex of the IFT B core. However, ift46, bld1, and ift88 mutants differ in phenotype as ift46 mutants are able to form short flagella, while the other two lack flagella completely. In this study, we investigated the functional differences of these individual IFT proteins contributing to complex B assembly, stability, and basal body localization. We found that complex B is completely disrupted in bld1 mutant, indicating an essential role of IFT52 for complex B core assembly. Ift46 mutant cells are capable of assembling a relatively intact complex B, but such complex is highly unstable and prone to degradation. In contrast, in ift88 mutant cells the complex B core still assembles and remains stable, but the peripheral proteins no longer attach to the B core. Moreover, in ift88 mutant cells, while complex A and the anterograde IFT motor FLA10 are localized normally to the transition fibers, complex B proteins instead are accumulated at the proximal ends of the basal bodies. In addition, in bld2 mutant, the IFT complex B proteins still localize to the proximal ends of defective centrioles which completely lack transition fibers. Taken together, these results revealed a step-wise assembly process for complex B, and showed that the complex first localizes to the proximal end of the centrioles and then translocates onto the transition fibers via an IFT88-dependent mechanism.

BACKGROUND: Intraflagellar transport (IFT) is the bidirectional movement of IFT particles between the cell body and the distal tip of a flagellum. Organized into complexes A and B, IFT particles are composed of at least 18 proteins. The function of IFT proteins in flagellar assembly has been extensively investigated. However, much less is known about the molecular mechanism of how IFT is regulated. METHODOLOGY/PRINCIPAL FINDINGS: We herein report the identification of a novel IFT particle protein, IFT25, in Chlamydomonas. Dephosphorylation assay revealed that IFT25 is a phosphoprotein. Biochemical analysis of temperature sensitive IFT mutants indicated that IFT25 is an IFT complex B subunit. In vitro binding assay confirmed that IFT25 binds to IFT27, a Rab-like small GTPase component of the IFT complex B. Immunofluorescence staining showed that IFT25 has a punctuate flagellar distribution as expected for an IFT protein, but displays a unique distribution pattern at the flagellar base. IFT25 co-localizes with IFT27 at the distal-most portion of basal bodies, probably the transition zones, and concentrates in the basal body region by partially overlapping with other IFT complex B subunits, such as IFT46. Sucrose density gradient centrifugation analysis demonstrated that, in flagella, the majority of IFT27 and IFT25 including both phosphorylated and non-phosphorylated forms are cosedimented with other complex B subunits in the 16S fractions. In contrast, in cell body, only a fraction of IFT25 and IFT27 is integrated into the preassembled complex B, and IFT25 detected in complex B is preferentially phosphorylated. CONCLUSION/SIGNIFICANCE: IFT25 is a phosphoprotein component of IFT particle complex B. IFT25 directly interacts with IFT27, and these two proteins likely form a subcomplex in vivo. We postulate that the association and disassociation between the subcomplex of IFT25 and IFT27 and complex B might be involved in the regulation of IFT.