TNFSF14 mediates the impact of docosahexaenoic acid on atopic dermatitis: a Mendelian randomization study.

OBJECTIVE: While current research suggests potential value for docosahexaenoic acid (DHA) in the prevention and management of atopic dermatitis (AD), the causal relationship between DHA and AD remains unclear, and the underlying mechanisms are not well understood. MATERIALS AND METHODS: To investigate the potential causal relationship between DHA and AD, as well as to explore potential mediating mechanisms, we employed the Mendelian randomization (MR) methods. To study these potential relationships, we conducted MR analysis using publicly available Genome-Wide Association Studies (GWAS) data. Effect estimates were computed using the random-effects inverse-variance weighted method. RESULTS: Our study demonstrates a negative correlation between DHA levels and AD risk (OR: 0.915, 95% CI: 0.858-0.975, p=0.007). Furthermore, in MR analysis using tumor necrosis factor ligand superfamily member 14 (TNFSF14) levels as an outcome, DHA levels also show a negative association with TNFSF14 levels (OR: 0.933, 95% CI: 0.879-0.990, p=0.022). Subsequently, we performed further analysis to explore the relationship between TNFSF14 and AD risk, revealing a positive correlation (OR: 1.069, 95% CI: 1.005-1.137, p=0.033). This suggests a potential mediating role of TNFSF14 in the impact of DHA on AD risk. CONCLUSIONS: In summary, our study employs MR analysis to offer genetic evidence indicating a potential role of DHA in reducing the risk of AD, as well as opening avenues for further in-depth investigation into potential mechanisms. These findings emphasize the importance of ongoing research in this field.

Enhancing Nasopharyngeal Carcinoma Cell Separation with Selective Fibronectin Coating and Topographical Modification on Polydimethylsiloxane Scaffold Platforms.

The extracellular matrix (ECM) serves as a complex scaffold with diverse physical dimensions and surface properties influencing NPC cell migration. Polydimethylsiloxane (PDMS), a widely used biocompatible material, is hydrophobic and undesirable for cell seeding. Thus, the establishment of a biomimetic model with varied topographies and surface properties is essential for effective NPC43 cell separation from NP460 cells. This study explored how ECM surface properties influence NP460 and NPC43 cell behaviors via plasma treatments and chemical modifications to alter the platform surface. In addition to the conventional oxygen/nitrogen (O2/N2) plasma treatment, O2 and argon plasma treatments were utilized to modify the platform surface, which increased the hydrophilicity of the PDMS platforms, resulting in enhanced cell adhesion. (3-aminopropyl)triethoxysilane and fibronectin (FN) were used to coat the PDMS platforms uniformly and selectively. The chemical coatings significantly affected cell motility and spreading, as cells exhibited faster migration, elongated cell shapes, and larger spreading areas on FN-coated surfaces. Furthermore, narrower top layer trenches with 5 µm width and a lower concentration of 10 µg/mL FN were coated selectively on the platforms to limit NP460 cell movements and enhance NPC43 cell separation efficiency. A significantly high separation efficiency of 99.4% was achieved on the two-layer scaffold platform with 20/5 µm wide ridge/trench (R/T) as the top layer and 40/10 µm wide R/T as the bottom layer, coupling with 10 µg/mL FN selectively coated on the sidewalls of the top and bottom layers. This work demonstrated an innovative application of selective FN coating to direct cell behavior, offering a new perspective to probe into the subtleties of NPC cell separation efficiency. Moreover, this cost-effective and compact microsystem sets a new benchmark for separating cancer cells.

Engineered barriers regulate osteoblast cell migration in vertical direction.

Considering cell migration is essential for understanding physiological processes and diseases. The vertical migration of cells in three dimensions is vital, but most previous studies on cell migration have only focused on two-dimensional horizontal migration. In this paper, cell migration in the vertical direction was studied. Barriers with a height of 1, 5, 10, and 25 µm with grating and arrows in channels as guiding patterns were fabricated. The effects of barrier height and guiding patterns on the vertical migration of MC3T3 cells were explored. The study revealed that taller barriers hinder vertical migration of MC3T3 cells, whereas grating and arrows in channels promote it. The time-lapse and micrograph images showed that as the barrier height increased, the cell climbing angle along the barrier sidewall decreased, and the time taken to climb over the barrier increased. These results indicate that taller barriers increase the difficulty of vertical migration by MC3T3 cells. To promote the vertical migration of MC3T3 cells, 10 µm tall barriers with 18° and 40° sloped sidewalls were fabricated. For barriers with 18° sloped sidewalls, the probability for MC3T3 cells to climb up and down the 10 µm tall barriers was 40.6% and 20.3%, respectively; this is much higher than the migration probability over vertical barriers. This study shows topographic guidance on the vertical migration of MC3T3 cells and broadens the understanding of cell migration in the vertical direction.

RAS and BRAF genes as biomarkers and target for personalised colorectal cancer therapy: An update.

Colorectal cancer (CRC) remains among the most commonly diagnosed cancers and has been on the rise. It is also one of the most lethal diseases globally, being the third leading cause of cancerrelated death. Depending on the stages and disease conditions, CRC is treated by surgery, chemo-, radio-therapy, immunotherapy or in combination. However, these therapies have subpar results with unwanted side effects, hence continuous effort is ongoing to explore new type of therapeutic modalities. Among the sub-types of CRC, KRAS, BRAF and NRAS mutated CRC comprise approximately 43%, 10% and 3% of the total cases, respectively. These mutations are associated with tumour progression and anti-epidermal growth factor receptor (EGFR) treatment resistance. Due to their important role in CRC, these genes have thus become targets in the development of novel treatments. In this paper, we discuss the current and upcoming treatment on CRC by targeting these mutated genes, with more focus on KRAS and BRAF due to the higher occurrence of mutations in CRC.

Separation of nasopharyngeal epithelial cells from carcinoma cells on 3D scaffold platforms.

Scaffold microstructures were developed to mimic a three-dimensional extracellular matrix in studying cell migration and invasion. The multiple-layer scaffold platforms were designed to investigate cell migration and separation from top to bottom layer. Two cell lines including immortalized nasopharyngeal epithelial (NP460) cells and nasopharyngeal carcinoma (NPC43) cells with Epstein-Barr virus were compared in this study. On one-layer platforms with trench depth of 15 µm, both NP460 and NPC43 cells were guided to migrate along the 18-µm-wide trenches, and exhibited random migration directions when the trench width was 10 or 50 µm. Nearly no cell was found to migrate in the 10-µm-wide trenches on one-layer platforms. However, the NP460 and NPC43 cells showed very different probability in the narrow trenches on two-layer platforms, making it possible to separate the nasopharyngeal epithelial cells from the carcinoma cells. Moreover, 1-µm deep grating topography on the top layer inhibited NP460 cells to migrate from top ridges to the 10-µm-wide trenches, but promoted such behavior for NPC43 cells. The results demonstrated in This study suggest that the engineered multiple-layer scaffold platforms could be used to separate carcinoma cells in NPC tumor as a potential treatment of NPC.

Effects of topographical guidance cues on osteoblast cell migration.

Cell migration is a fundamental process that is crucial for many biological functions in the body such as immune responses and tissue regeneration. Dysregulation of this process is associated with cancer metastasis. In this study, polydimethylsiloxane platforms with various topographical features were engineered to explore the influence of guiding patterns on MC3T3-E1 osteoblast cell migration. Focusing on the guiding effects of grating patterns, variations such as etch depth, pattern discontinuity, and bending angles were investigated. In all experiments, MC3T3-E1 cells on patterned surfaces demonstrated a higher migration speed and alignment when compared to flat surfaces. The study revealed that an increase in etch depth from 150 nm to 4.5 µm enhanced cell alignment and elongation along the grating patterns. In the presence of discontinuous elements, cell migration speed was accelerated when compared to gratings of the same etch depth. These results indicated that cell directionality preference was influenced by a high level of pattern discontinuity. On patterns with bends, cells were more inclined to reverse on 45° bends, with 69% of cells reversing at least once, compared to 54% on 135° bends. These results are attributed to cell morphology and motility mechanisms that are associated with surface topography, where actin filament structures such as filopodia and lamellipodia are essential in sensing the surrounding environment and controlling cell displacement. Knowledge of geometric guidance cues could provide a better understanding on how cell migration is influenced by extracellular matrix topography in vivo.

Traversing behavior of tumor cells in three-dimensional platforms with different topography.

Three-dimensional polydimethylsiloxane platforms were developed to mimic the extracellular matrix with blood vessels by having scaffolds with micropatterns, porous membrane and trenches. Precisely controlled physical dimensions, layouts, and topography as well as different surface chemical treatments were applied to study their influences on nasopharyngeal carcinoma cell (10-15 µm in diameter) migration in mimicked platforms over 15-hour of time-lapse imaging. By placing the pores at different distance from the edges of the trenches, pores with different trench sidewall exposures and effective sizes were generated. Pores right next to the trench sidewalls showed the highest cell traversing probability, most likely related to the larger surface contact area with cells along the sidewalls. Straight grating oriented perpendicular to trenches below the top layer increased cell traversing probability. Pore shape as well as pore size influenced the cell traversing probability and cells could not traverse through pores that were 6 µm or less in diameter, which is much smaller than the cell size. Trench depth of 15 µm could induce more cells to traverse through the porous membrane, while shallower trenches impeded cell traversing and longer time was needed for cells to traverse because 3 and 6 µm deep trenches were much smaller than cell size which required large cell deformation. Hydrophobic surface coating on the top layer and fibronectin in pores and trenches increased the cell traversing probability and reduced the pore size that cells could traverse from 8 to 6 µm, which indicated that cells could have larger deformation with certain surface coatings.

Partial thyroxine-binding globulin deficiency in a family with coding region mutations in the TBG gene.

PURPOSE: T4-binding globulin (TBG) is the main thyroid hormone (TH) transporter present in human serum. Inherited thyroxine-binding globulin (TBG) deficiency is caused by mutations in the TBG (SERPINA7) gene, which is located on the X chromosome. This study was performed to report and evaluate coding region mutations in TBG gene for partial thyroxine-binding globulin deficiency. METHODS: A pedigree spanning four generations is described in this study. The proband is a female with partial TBG deficiency. All members of this pedigree underwent thyroid function tests, while Sanger sequencing was used to identify the TBG gene mutations. Bioinformatics databases were used to evaluate the deleterious effects of the mutation(s). Two hundred and seven unrelated individuals were used to evaluate the thyroid function of individuals with different TBG mutations. A one-way ANOVA was used to analyze the impact of the TBG mutations on thyroid function. RESULTS: TBG gene sequencing results revealed that the proband had a novel mutation in codon 27 leading to alanine to valine substitution (p.A27V). This mutation was associated with lower serum T4 levels (p < 0.0001) when compared to the groups that did not carry the mutation. The previously reported p.L283F mutation was also found in the proband. The hemizygous p.L283F individuals presenting with lower T4 serum and TBG levels (p < 0.001) when compared to wildtype males and females. Both mutations were deleterious upon SIFT and PolyPhen-2 evaluation. CONCLUSION: Associated with partial thyroxine-binding globulin deficiency, this study reports a novel p.A27V mutation in the TBG gene.

Integration of biochemical and topographic cues for the formation and spatial distribution of invadosomes in nasopharyngeal epithelial cells.

Invadosomes are invasive protrusions generated by cells which can secrete matrix metalloproteinases for focal digestion of extracellular matrix. They also aid invasive cancer cells in their transmigration through vascular endothelium. However, how the physical and chemical cues in a three-dimensional (3D) system signal the spatial localization of invadosomes remains largely unknown. Here we study the topographic guidance of invadosome formation in invasive nasopharyngeal cells under the stimulation of an inflammatory cytokine, TGF-ß1, using engineered gratings with different width and depth. We first report that TGF-ß1 can act as an external signal to upregulate the formation of invadosomes with a random distribution on a plane 2D surface. When the cells were seeded on parallel 3D gratings of 5 µm width and 1 µm depth, most of the invadosomes aligned to the edges of the gratings, indicating a topographic cue to the control of invadosome localization. While the number of invadosomes per cell were not upregulated when the cells were seeded on 3D topography, guidance of invadosomes localization to edges is correlated with cell migration directionality on 1 µm deep gratings. Invadosomes preferentially form at edges when the cells move at a lower speed and are guided along narrow gratings. The invadosomes forming at 3D edges also have a longer half-life than those forming on a plane surface. These data suggest that there are integrated biochemical and 3D geometric cues underlying the spatial regulation of invasive structures so as to elicit efficient invasion or metastasis of cells. STATEMENT OF SIGNIFICANCE: Nasopharyngeal cells were integrated with the biological cues and matrix topography to govern the activity and spatial distribution of invadosomes. The biochemical induction of invadosome formation by TGF-ß1 in nasopharyngeal cells was observed. When the cells were seeded on parallel 3D gratings, most of the invadosomes aligned to the edges of the gratings due to topographical induced invadosome localization. While the number of invadosomes per cell were not upregulated, guidance of invadosomes localization to edges is correlated with cell migration directionality on 1 µm deep gratings. Invadosomes preferentially form at edges with a higher stability when the cells are guided along narrow gratings. The integrated biochemical and 3D geometric cues could elicit efficient invasion or metastasis of cells.

Pulmonary Embolism as a Cause of Death in Psychiatric Inpatients: a Case Series.

We report four cases of fatal pulmonary embolism confirmed by autopsy among inpatients in a Hong Kong psychiatric hospital from 2010 to 2014. None of the four patients had a medical or premorbid condition associated with vascular thromboembolism or causing prolonged immobilisation. Only two patients were taking long-term antipsychotic medication, but all were physically restrained shortly before the event.

Control of neural probe shank flexibility by fluidic pressure in embedded microchannel using PDMS/PI hybrid substrate.

Implantable neural probes are widely used to record and stimulate neural activities. These probes should be stiff enough for insertion. However, it should also be flexible to minimize tissue damage after insertion. Therefore, having dynamic control of the neural probe shank flexibility will be useful. For the first time, we have successfully fabricated flexible neural probes with embedded microfluidic channels for dynamic control of neural probe stiffness by controlling fluidic pressure in the channels. The present hybrid neural probes consisted of polydimethylsiloxane (PDMS) and polyimide (PI) layers could provide the required stiffness for insertion and flexibility during operation. The PDMS channels were fabricated by reversal imprint using a silicon mold and bonded to a PI layer to form the embedded channels in the neural probe. The probe shape was patterned using an oxygen plasma generated by an inductively coupled plasma etching system. The critical buckling force of PDMS/PI neural probes could be tuned from 0.25-1.25 mN depending on the applied fluidic pressure in the microchannels and these probes were successfully inserted into a 0.6% agarose gel that mimicked the stiffness of the brain tissue. Polymer-based neural probes are typically more flexible than conventional metal wire-based probes, and they could potentially provide less tissue damage after implantation.

Dynamic Tracking of Osteoblastic Cell Traction Force during Guided Migration.

INTRODUCTION: Continuous development of cell traction force can regulate cell migration on various extracellular matrixes in vivo. However, the topographical effect on traction force is still not fully understood. METHODS: Micropost sensors with parallel guiding gratings were fabricated in polydimethylsiloxane to track the cell traction force during topographical guidance in real time. The force distributions along MC3T3-E1 mouse osteoblasts were captured every minute. The traction force in the leading, middle, and trailing regions was monitored during forward and reversed cell migration. RESULTS: The traction force showed periodic changes during cell migration when the cell changed from elongated to contracted shape. For cell migration without guiding pattern, the leading region showed the largest traction force among the three regions, typically 5.8 ± 0.8 nanonewton (nN) when the cell contracted and 7.1 ± 0.5 nN when it elongated. During guided cell migration, a lower traction force was obtained. When a cell contracted, the trailing traction force was 4.1 ± 0.4 for non-guided migration and 2.2 ± 0.2 nN for guided migration. As a cell became elongated, the trailing traction force was 6.0 ± 0.5 nN during non-guided migration and 4.8 ± 0.3 nN under guidance. When a cell reversed its migration direction, the magnitudes of the traction force from the leading to the trailing regions also flipped. CONCLUSION: The cell traction force is continuously influenced by topographical guidance, which determines cell migration speed and direction. These results of cell traction force development on various topographies could lead to better cell migration control using topotaxis.

A microfabricated low-profile wideband antenna array for terahertz communications.

While terahertz communications are considered to be the future solutions for the increasing demands on bandwidth, terahertz equivalents of radio frequency front-end components have not been realized. It remains challenging to achieve wideband, low profile antenna arrays with highly directive beams of radiation. Here, based on the complementary antenna approach, a wideband 2 × 2 cavity-backed slot antenna array with a corrugated surface is proposed. The approach is based on a unidirectional antenna with a cardiac radiation pattern and stable frequency characteristics that is achieved by integrating a series-resonant electric dipole with a parallel-resonant magnetic dipole. In this design, the slots work as magnetic dipoles while the corrugated surface radiates as an array of electric dipoles. The proposed antenna is realized at 1 THz operating frequency by stacking multiple metallized layers using the microfabrication technology. S-parameter measurements of this terahertz low-profile metallic antenna array demonstrate high efficiency at terahertz frequencies. Fractional bandwidth and gain are measured to be 26% and 14 dBi which are consistent with the simulated results. The proposed antenna can be used as the building block for larger antenna arrays with more directive beams, paving the way to develop high gain low-profile antennas for future communication needs.

The influence of precompression on elasticity of thyroid nodules estimated by ultrasound shear wave elastography.

OBJECTIVES: To investigate the influence of variations in resting pressure (precompression) on thyroid ultrasound supersonic shear wave elastography (SWE). METHODS: Thirty-five normal thyroid glands (Norm), 55 benign hyperplastic nodules (BHN), and 17 papillary thyroid cancers (PTC) in 96 subjects underwent thyroid SWE. Four precompression levels were applied manually by the operator, ranging from A (baseline, 0 % strain) to D (high, 22-30 % strain). SWE results at each precompression level were compared using ANOVA tests with P < 0.05 indicating significance. RESULTS: SWE indices were highest in PTC, followed by BHN and Norm at each precompression level (P < 0.05). All tissue types showed successive increases in SWE results as precompression increased, although the rate was higher for PTC than BHN and Norm (Ps < 0.05). SWE values (kPa) of Norm, BHN, and PTC at baseline precompression (A) were 10.3 ± 3.3, 17.7 ± 7.6, and 22.2 ± 11.9 compared with 21.1 ± 4.2, 42.3 ± 16.0, and 97.6 ± 46.8 at high precompression (D). SWE index differences between precompression levels A and D were 10.8 kPa for Norm, 24.6 kPa for BHN, and 75.4 kPa for PTC. CONCLUSION: PTCs show greater SWE stiffening than BHN as precompression rises. Precompression effects on thyroid nodules are not negligible and may account for wide discrepancies in published SWE discriminatory performance results for thyroid malignancy. KEY POINTS: • Increases in resting pressure (precompression) applied by the operator increases thyroid stiffness. • Papillary cancers show greater increases in stiffness (strain hardening) than benign nodules. • Precompression may affect the diagnostic performance of shearwave elastography for thyroid malignancy.

Electrode modifications to lower electrode impedance and improve neural signal recording sensitivity.

OBJECTIVE: Although electrode size should be miniaturized to provide higher selectivity for neural signal recording and to avoid tissue damage, small sized electrodes induce high impedance, which decreases recording quality. In this work, the electrode surface was modified to increase the effective surface area to lower the electrode impedance and to improve the neural signal detection quality by optimizing plasma conditions. APPROACH: A tetrafluoromethane (CF4) plasma was used to increase the effective surface area of gold electrode sites of polyimide-based neural probes. In vitro electrode impedance and in vivo neural signal recording and stimulation were characterized. MAIN RESULTS: For 15 µm diameter (dia.) electrode size, the average surface roughness could be increased from 1.7 to 22 nm after plasma treatment, and the electrode impedance was decreased by 98%. Averaged background noise power in the range of 1 to 1000 Hz was decreased to -106 dB after the 30 µm dia. electrodes were plasma modified-lower than the noise level of -86 dB without plasma treatment. Neural probes with plasma-modified electrode sites of 15 and 30 µm dia. were implanted to the anterior cingulate cortex (ACC) region for acute recording of spontaneous and electrical evoked local field potential (LFP) of neural signals. Spontaneous LFP recorded in vivo by the plasma-modified electrodes of 30 µm dia. was two times higher compared to electrodes without treatment. For a stimulation current of 400 µA, electrically evoked LFP recorded by the plasma-modified electrodes was seven times higher than those without plasma exposure. SIGNIFICANCE: A controllable technology was developed to increase the effective surface area of electrodes using a CF4 plasma. Plasma-modified electrodes improved the quality of the neural probe recording and more sensitive to record spontaneous and evoked LFP in the ACC region.

Control of cell migration direction by inducing cell shape asymmetry with patterned topography.

In this study, we explored the concept of introducing asymmetry to cell shapes by patterned cell culture substrates, and investigated the consequence of this induced asymmetry to cell migration behaviors. Three patterns, named "Squares", "Grating", and "Arcs" were fabricated, representing different levels of rotational asymmetry. Using time-lapse imaging, we systematically compared the motility and directionality of mouse osteoblastic cells MC3T3-E1 cultured on these patterns. Cells were found to move progressively faster on "Arcs" than on "Grating", and cells on "Squares" were the slowest, suggesting that cell motility correlates with the level of rotational asymmetry of the repeating units of the pattern. Among these three patterns, on the "Arcs" pattern, the least symmetrical one, cells not only moved with the highest velocity but also the strongest directional persistence. Although this enhanced motility was not associated with the detected number of focal adhesion sites in the cells, the pattern asymmetry was reflected in the asymmetrical cell spreading. Cells on the "Arcs" pattern consistently displayed larger cytoplasmic protrusion on one side of the cell. This asymmetry in cell shape determined the direction and speed of cell migration. These observations suggest that topographical patterns that enhance the imbalance between the leading and trailing fronts of adherent cells will increase cell speed and control movement directions. Our discovery shows that complex cell behaviors such as the direction of cell movement are influenced by simple geometrical principles, which can be utilized as the design foundation for platforms that guide and sort cultured cells.

The immobilization of DNA molecules to electrodes in confined channels at physiological pH.

Large numbers of DNA molecules are immobilized to electrodes at the physiological pH of 8.0, and the length of the immobilized DNA molecules is controlled using an ac voltage. Efficient DNA immobilization at physiological pH has been demonstrated by integrating electrodes in confined channels 500 nm wide and 100 nm deep. The low volume of the channels allows large numbers of DNA molecules to access the electrode surfaces, leading to efficient immobilization.

Effects of nanoimprinted patterns in tissue-culture polystyrene on cell behavior.

Tissue engineering seeks to develop functional tissues in a biomimetic environment in vitro. As the extracellular environment in vivo is composed of numerous nanostructures, fabrication of nanostructured substrates will be valuable for tissue engineering applications. In this article, we report a simple nanoimprint lithography (NIL) process to pattern nanostructures directly on tissue-culture polystyrene plates. By repeating this NIL process, three-dimensional scaffolds consisting of multiple-layer nanostructures were also fabricated. Bovine pulmonary artery smooth muscle cells were cultured on imprinted gratings ranging from 350 nm to 10 µm. The smooth muscle cells attached and proliferated well on these imprinted substrates without additional surface treatment. Cell elongation and alignment were observed on the micro- and nanopatterns, with the effect significantly more pronounced on the nanostructures.

Nasopharyngeal intraepithelial lesion: latent Epstein-Barr virus infection with malignant potential.

AIMS: To study the morphology and immunohistochemical expression of nasopharyngeal intraepithelial lesions and to understand their place in nasopharyngeal carcinogenesis. METHODS AND RESULTS: Nine cases of nasopharyngeal intraepithelial lesion (NPIL) were diagnosed during nasopharyngeal biopsy screening for nasopharyngeal carcinoma (NPC). Two cases were associated with early invasion. All cases demonstrated specific histological features and consistent positivity on in-situ hybridization for Epstein-Barr virus (EBV)-encoded RNA. Pure NPIL lesions showed low-grade morphology while lesions associated with early invasion were high grade. Immunohistochemical studies showed increased expression of bcl-2 and essentially negative findings for BZLF1 and LMP1. High-grade lesions had relatively stronger expression of bcl-2 and p53. CONCLUSIONS: NPIL harbours latent EBV infection and has malignant potential. Multiple steps are involved in its occurrence and progression. Low-grade and high-grade lesions should be managed differently.