Evaluation of perioperative pachymetry of thin corneas during corneal crosslinking using hypo-osmolar riboflavin and hydroxypropyl methylcellulose

ABSTRACT Purpose: This study aimed to analyze variations in intraoperative corneal thickness during corneal cross-linking in patients with keratoconus and to investigate its possible correlation with presurgical maximal keratometry (Kmax) and pachymetry. Methods: This was a prospective case series. We used a method similar to the Dresden protocol, with the application of hydroxypropyl methylcellulose 0.1% hypo-osmolar riboflavin in corneas between 330 and 400 µm after epithelium removal. Corneal thickness was measured using portable calipers before and immediately after epithelium removal, and 30 and 60 min after the procedure. Results: The 30 patients in this study were followed up for one year. A statistically significant difference was observed in pachymetry values during the intraoperative period (p<0.0001) and an increase of 3.05 µm (95%C1: 0.56-5.54) for each diopter was seen after epithelium removal (p0.019). We found an average Kmax difference of —2.12 D between men and women (p0.013). One year after treatment, there was a statistically significant reduction in pachymetry (p<0.0001) and Kmax (p0.0170) values. Conclusions: A significant increase in pachymetry measurements was seen during the procedure, and most patients showed a regression in Kmax and pachymetry values one year after surgery.

Effect of short-term exposure to Raag Bilawal of North Indian classical music on young Indian adults: a high-density electroencephalogram microstate study.

Introduction: the objective of the study was to find out the microstate map topographies and their parameters generated during the resting state and during listening to North Indian classical Music Raag 'the Raag Bilawal'. It was hypothesized that in the resting state and during listening to music conditions, there would be a difference in microstate parameters i.e. mean duration, global explained variance (GEV), and time coverage. Methods: a 128-channel electroencephalogram (EEG) was recorded for 12 Indian subjects (average age 26.1+1.4 years) while resting and listening to music using the EEG microstate investigation. Investigation and comparison of the microstate parameters were the mean duration, global explained variance (GEV), and time coverage between both conditions were performed. Results: seven microstate maps were found to represent the resting state and listening to music condition, four canonical and three novel maps. No statistically significant difference was found between the two conditions for time coverage and mean duration. The statistical significance levels of the map-1, map-2, map-3, map-4, map-5, map-6, and map-7 for the mean duration were 0.4, 0.6, 0.97, 0.34, 0.32, 0.69, and 0.29 respectively; and for time coverage were 0.92, 0.92, 0.96, 0.64, 0.78, 0.38, and 0.76 respectively. Map-1, map-4, and map-7 were the three novel maps we found in our study. Conclusion: similarities regarding stability and predominance of maps with small vulnerability exist in both conditions indicating that phonological, visual, and dorsal attention networks may be activated in both resting state and listening to music condition.

Effect of micropillar density on morphology and migration of low and high metastatic potential breast cancer cells.

Study of cell migration in cancer is crucial to the comprehension of the processes and factors that govern tumor spread. Cancer cells migrate invading tissues, causing alterations in cell adhesion, cytoskeleton, and signaling pathways. Little is known about the physical attributes of cancer cells that change when interacting with microenvironments. In this work, the local topography of the ECM has been mimicked through micropillar array substrates. MDA-MB-231 and MCF-7 breast cancer cells, exhibiting high and low metastatic potential, respectively, were analyzed. Differences in morphology and migration of the cells were investigated by examining the cell spreading area, circularity, aspect ratio, migration speed, and migration path. This work encountered that none of the studied cell lines have preferential orientation migrating on uniform patterns. In contrast, cell migration on graded patterns shows preferential orientation along the longitudinal direction from sparser to denser zones which is significantly influenced by substrate stiffness and indicates that both cell lines can sense the spacing gradient and respond to this topographical cue. The migration speed of the breast cancer cell lines significantly decreases from the sparse to medium to dense zones, registering higher values for the MDA-MB-231.

Medicalizing the Body and the Locale: Kala Azar and Disease Thinking in Assam, 1824-1900.

The article examines two seemingly unconnected occurrences at the nineteenth-century north east frontier of British India. The first is the production of a pathological space via moral, social, and cultural codes enacted by medical topographies on the region since the first Anglo-Burmese war (1824-1826) and the subsequent rise of disease thinking. The second is the ambivalence in disease thinking that is brought to fore through the mysterious malady called kala azar (visceral leishmaniasis), which was geographically designated as Assam fever. This article contends that the geographical designation of kala azar as Assam fever is not just coincidental or a nosological confusion of the late nineteenth century but rather has its origin in the preceding pathological carving of space at the frontier. Further, it explores the troubled ontology between research on malaria and kala azar investigations to show that the old codes enacted by medical topographies hinged upon the era of laboratory medicine.

Coolants, organic acids, flavourings and other additives that facilitate inhalation of tobacco and nicotine products: implications for regulation.

To inform regulatory policy, this article summarises findings on inhalation facilitation from the ninth report of the WHO Study Group on Tobacco Product Regulation. Some additives counteract the harshness and bitterness of tobacco and nicotine product aerosols, making them easier to inhale. Additives that promote inhalability may perpetuate and increase the use of inhaled tobacco and nicotine products, especially by young people. Thus, as a class, additives that facilitate inhalation are an important regulatory target to prevent tobacco and nicotine product uptake. We defined inhalation facilitation as modifications to products during manufacturing that enhance the sensory experience and (potentially) behaviours associated with inhalation (eg, deeper puffs, faster inhalation, larger puff volume, shorter intervals in between puffs and use episodes). Evidence review showed that: (a) menthol and synthetic coolants decrease irritation caused by aerosol constituents by activating sensory perception receptors (eg, cooling receptors) and may promote dependence in inexperienced users; (b) acid additives and sugars, which lower the pH of aerosols and shift nicotine from free-base to protonated salt forms, reduce harshness and increase blood nicotine yield; (c) e-cigarette flavourings perceived as sweet or fruity reduce subjective bitterness, increase attractiveness and may escalate use, although their effects on perceived harshness are inconclusive; (d) sugars in tobacco impart sweet sensations, but limited industry-independent data preclude strong conclusions for sugars' roles in inhalation facilitation. Given these findings, WHO policy recommendations suggest that regulators might consider banning ingredients that facilitate inhalation in all commercial inhaled tobacco and nicotine products.

Tectonic structures of the Dome Fuji region, East Antarctica, based on new magnetic data.

The Oldest Ice Reconnaissance (OIR) airborne geophysical survey in East Antarctica was flown over approximately 170,000 km2 of the Dome Fuji region in 2016/17. The survey's results support new insights into the subglacial geology and its meaning for the tectonic histories of the supercontinents Rodinia and Gondwana. The new magnetic and radar-derived bed topography data are integrated with previously acquired magnetic and gravity data, allowing the mapping of crustal domains within and beyond the survey's limits. The magnetic data reveal three distinct domains within the survey region, delineated by N-S oriented boundaries, partly aligned with gravity domains following upward continuation transformations for both datasets. Additionally, four primary sets of magnetic lineaments were identified, exhibiting correlations with topographic and gravity patterns. These correlations indicate the continuation of the Tonian Oceanic Arc Super Terrane (TOAST) southward of its previously known southern limit. Moreover, an E-W-trending magnetic anomaly, the Elbert magnetic anomaly, suggests the suture between the recently-proposed subglacial Valkyrie craton and the TOAST. Furthermore, the analysis reveals a broad scale shear zone, named here the OIR shear zone, which formed as a result of oblique collision of the Ruker and Valkyrie cratons during the amalgamation of Gondwana.

Pear-Shaped Craniopharyngioma: Selection of the Transcranial Approach With Respect to Topographical Classifications.

We present the selection of the transcranial microsurgical approach, operative treatment, and outcomes following the resection of a pear-shaped craniopharyngioma (CP). A nine-year-old boy was operated on and followed up for 2.5 years after radical resection of the extrapial CP. Postoperatively, there was no tumor recurrence. The surgical strategy was discussed based on the preoperative MRI appearance of the CP, especially its morphological characteristics, including not only its size and shape but also its relationship with the hypothalamus, pituitary stalk/gland, ventricles, and optic chiasm, and the possible location of perforators. A description of the tumor topography is provided together with a discussion on the rationale for the selection of our surgical approach. Based on an understanding of the tumor topography, important information can be gained for approach selection, surgical planning, and anticipation of the hypothalamic-pituitary outcome.

Medicalizing Topography: The French Army and Mediterranean Climates, 1760-1860.

This article details how the French army employed medical topography as a tool of military occupation throughout the Mediterranean world from the mid-eighteenth to mid-nineteenth century. It departs from other works by focusing exclusively on medical topography's military applications. Medical topographies charted the connections between health and the environment by observing a location's features, such as soil, air, and water quality, as well as elevation, prevailing winds, common local diseases, sources of potential contagion, and the cleanliness of urban environments. Because a medical-topographic study took time to write and implement, its findings provided little utility during active conflict. Only after the fighting ceased during a campaign could the army make use of a medical topography's findings by taking measures such as draining swamps, relocating hospitals in unhealthy environments, and issuing climate-appropriate gear.

Cet article examine la façon dont l'armée française utilisait la topographie médicale en tant qu'outil d'occupation militaire pendant les dix-huitième et dix-neuvième siècles dans le monde Méditerranéen. Il se détache des autres travaux en se concentrant exclusivement sur les applications militaires de l'étude. Les études topographiques-médicales analysaient des liens entre l'environnement et la santé. Ces études permettaient l'observation minutieuse et l'enregistrement des caractéristiques d'un lieu, comme par exemple : son élévation, les vents dominants, la qualité de terre, d'eaux, et d'air, la propreté des centres urbaines, et des maladies locales ainsi que leurs origines. Parce que ces études exigeaient du temps d'écrire et d'implémenter, elles n'ont pas eu une grande utilité pratique pendant une campagne. Cependant, après la cessation des hostilités, l'armée a mis en pratique les résultats de ces études. Elle a vidé des marais, déménagé des hôpitaux malsains, et distribué du matériel adapté à l'environnement.

Morphometry and skeletopy of kidneys and renal vessels in marmoset (Callithrix spp.).

BACKGROUND: Marmosets, Callithrix spp, are small New World monkeys that have gained importance as an experimental animal model for human. Despite its use, information on its renal morphometry, vascularization, and location are limited. Therefore, this study will supply basic anatomy for applied studies and for comparative anatomy. METHODS: Fifty cadavers of Callithrix spp were collected on highways from the Atlantic Forest biome, identified and injected with a 10% formaldehyde solution. Later, the specimens were dissected and the measurements and topography of the kidneys and renal vessels were recorded. Both left and right kidneys were significantly larger in females. RESULTS: In the specimens studied, the average body length was 20.00 ± 2.46 cm in males and 20.50 ± 1.98 cm in females (p = .43). The kidneys of the Callithrix spp. were symmetrical in shape and resembled a "bean." They were also pale brown with a smooth surface. In males, the most frequent location of the right kidney was at the L1-L2 level (92%), while the location of the left kidney was between L2 and L3 (76%). In females, the most frequent location of the right kidney was at the L1-L2 level (56%), while the location of the left kidney was between L2 and L3 (32%) (Table 1). However, in seven (28%) males and nine (36%) females, the kidneys were at the same level. CONCLUSIONS: In both sexes, there was a positive and significant linear correlation between body length and kidney length. Regardless of the variable location of the kidneys in both sides and in either sexe, the right kidney was always located more cranially than the left, similar to observations in other non-human primates.

Clinical outcomes after topography-guided FS-LASIK for myopia with nonastigmatic eyes.

BACKGROUND: To analyze the clinical outcomes after topography-guided femtosecond laser-assisted laser in situ keratomileusis (FS-LASIK) with Phorcides Analytic Engine (PAE) algorithm or Custom-Q FS-LASIK for myopia with nonastigmatic eyes. METHODS: In this retrospective study, a total of 90 eyes with myopia without manifest astigmatism (82 patients) were included. All surgeries were performed by topography-guided FS-LASIK planned with a PAE algorithm (42 eyes) or Custom-Q system (48 eyes). Refractive, visual outcomes and corneal aberrations were compared between the two groups. RESULTS: At 6 months postoperatively, the postoperative uncorrected distance visual acuity (UDVA) was 20/20 or better in 42 eyes (100%) in the PAE compared with 44 eyes (92%) in Custom-Q (P = .120). The postoperative UDVA of 20/16 or better was measured in 92% of eyes in the PAE group and 81% of eyes in the Custom Q group (P = .320). Postoperative corrected distance visual acuity, manifest refractive spherical equivalent and refractive astigmatism were similar between the two groups (P > .05). The postoperative optical path difference (OPD) and Strehl ratio (SR) were significantly better in the PAE group compared with the Custom Q group. CONCLUSIONS: Topography-guided FS-LASIK with PAE algorithm or Custom Q demonstrated similar refractive efficacy and predictability. PAE for the patients with zero manifest astigmatism demonstrated better results in correcting corneal aberrations.

Improving the identification of pollution source areas with catchment-resolution sensitivity analysis.

The significant impacts of total nitrogen (TN) and total phosphorus (TP) on riverine ecosystems underscores the critical need to identify the primary nutrient source areas in watersheds. This study aims to unravel the influences of terrain and land use types on mean monthly TN (TNM) and mean monthly TP (TPM) export across varying catchment resolutions in the Qiantang River Watershed of China. The findings of this study illuminated the critical role of topography in understanding nutrient dynamics, wielding a profound influence over water flow patterns and nutrient dispersion. Both land slope and Stream Power Index (SPI) displayed substantial negative correlations (r < -0.6) with TNM and TPM concentrations, whereas the Topographic Wetness Index (TWI) showed positive correlations with the nutrient indexes. In addition to terrain characteristics, impervious land surfaces had a positive correlation with nutrient concentrations, while grassland and forest areas exhibited negative correlations. Results further underscored the substantial influence of catchment resolution on correlations between watershed properties and riverine nutrient concentrations. It was imperative to choose an effective catchment resolution in watershed delineation - not too coarse, nor too fine - to accurately capture the topographic and land use impacts on nutrient dynamics. With the most appropriate catchment size (Catchment 700 km2), the critical pollution source areas for TN and TP pollution were identified, and thus could be used to guide future pollution reduction efforts. The study not only highlights the importance of identifying an appropriate catchment size for water pollution, but also emphasizes the necessity of effectively extracting critical pollution source areas to mitigate water nutrient pollution and increase the ecological integrity of the Qiantang River Watershed.

Corneal aberrations are associated with low-energy meniscus injuries.

BACKGROUND: Visual impairment can cause balance problems. Therefore, visual impairment caused by an increase in corneal deviations can lead to sudden and unstable loads in the lower extremities. We aimed to investigate the possible relationship between low-energy meniscal injuries and corneal structural measures. METHODS: This prospective, observational study included individuals aged between 18-40 years with a normal body-mass index. The study group consisted of 54 patients with grade 2 or 3 meniscus injuries after low-energy activity. The control group consisted of 54 healthy individuals without any complaints in the knee joint. The corneal parameters of all participants were evaluated with a Scheimpflug corneal topography and specular microscopy device. Simulated keratometry (SimK), minimum central corneal thickness (MCCT), cylindrical diopter (ClyD), corneal volume (CVol) spheric aberrations (SphAbb), high-order aberration (HOA), coma values, and endothelial parameters were recorded. RESULTS: The research and control groups were similar in terms of age, body mass index, and gender distribution. There was no significant difference between the groups in the corneal SimK and CylD, parameters. However, HOA, Coma, SphAbb, and cell variability (Cv) values were significantly higher in the study group, and contrarily MCCT, CVol, and endothelial count (Cd) values were significantly lower. CONCLUSIONS: Our findings suggest that individuals with relatively lower MCCT values tend to develop meniscal damage after low-energy activity. Hence, the loss of corneal strength in these patients may be a sign of possible weakness in the meniscus. The HOA value above 0.26, the coma value above 0.16, and the SphAbb value above 0.1 may significantly increase the possible meniscus injury.

Bubble-graft coverage after Descemet Membrane Endothelial Keratoplasty depending on gaze angle.

PURPOSE: In Descemet Membrane Endothelial Keratoplasty (DMEK) a gas bubble is usually used to attach the graft to the host cornea. In this study, we observed the bubble size and bubble-graft coverage at different gaze angles following DMEK. METHODS: This observational prospective study analyzed 465 images of patients who underwent an uneventful DMEK. Intraoperatively, the anterior chamber was filled up to 90% of its volume with a 20% Sulfur Hexafluoride (SF6) gas-air mixture. Postoperatively, the bubble was photographed daily in different gaze angles ranging from a supine position (0°) to an upright position (90°) and a slightly inclined position (105°). The primary outcomes were bubble-graft coverage and bubble diameters depending on the gaze angle and time after DMEK. RESULTS: The highest bubble-graft coverage was achieved at a 0° gaze angle at all times of measurement. In the first 48 h after DMEK, the mean bubble-graft coverage was over 85% at a gaze angle between 0° and 45°. Starting 72 h after DMEK, the graft coverage declined at all gaze angles. The graft coverage at a 0° gaze angle was 88.61 ± 10.90% after 96 postoperative hours, while the graft coverage was below 85% at all other gaze angles. CONCLUSION: Our clinical results provide novel insight into variation in bubble-graft coverage as a function of gaze angle and may be used to aid in patient counselling for appropriate body positioning following DMEK to prevent early graft detachment. Maintaining supine positioning seems to be most advantageous starting 48 h after DMEK.

Higher Order Aberrations in Patients with Unilateral Nasolacrimal Duct Obstruction.

Purpose: To compare corneal higher order aberrations (HOAs) in patients with epiphora caused by unilateral primary acquired nasolacrimal duct obstruction (PANDO) and their fellow eyes. Methods: One hundred twenty two eyes of 61 patients with unilateral PANDO were included. Affected eyes named as Group 1 and contralateral eyes named as Group 2. Munk scale and fluorescein dye disappearance test (FDDT) were assessed. Corneal topographic and HOAs measurements were taken by Sirius topography. Results: There were 47 female and 14 male patients. The mean Km were 43.84± 1.4 in Group 1 and 43.96± 1.4 in Group 2 (p=0.73). The mean corneal topographic astigmatism were 0.81± 0.7 in Group 1 and 0.78± 0.5 in Group 2 (p=0.57). The mean coma-like aberration was 0.19± 0.2 in Group 1 and 0.1± 0.1 in Group 2 (p=0.03). The mean spherical-like aberration was 0.16± 0.1 in Group 1 and 0.11± 0.1 in Group 2 (p=0.04). The mean total corneal HOA was 0.3±0.5 in Group 1 and 0.23±0.2 in Group 2 (p=0.04). The mean munk score was 3.47± 1 in Group 1. There was a positive correlation between munk score and coma-like aberration and total corneal HOAs levels (r=0.53; p=0.003 and r=0.44; p=0.01 respectively). The mean FDDT was grade 2.6± 0.1 in Group 1. There was a positive correlation between FDDT grade and coma-like aberration and total corneal HOAs levels (r=0.48; p=0.001 and r=0.33; p=0.001 respectively). Conclusions: Epiphora in patients with PANDO can affect corneal HOAs. As patients symptoms and tear pooling were increased corneal HOA levels increased.

Swept-source optical coherence tomography in ocular surface diseases: anterior segment analysis repeatability and its limits.

Purpose: This study aims to evaluate the repeatability of anterior segment optical coherence tomography (AS-OCT) in diverse ocular surface disorder (OSD) cohorts, exploring various anterior segment parameters and their accuracy in different disease groups. Methods: A total of 239 participants across six distinct OSD groups and healthy controls underwent nonmydriatic AS-OCT imaging using the Tomey CASIA 2 device. Anterior segment parameters including anterior chamber depth, width, angle metrics, corneal thickness, keratometry, lens vault, and others were meticulously assessed. Statistical analyses determined repeatability limits and coefficients of variation for each parameter within the different OSD cohorts. Results: Repeatability for anterior chamber and corneal parameters remained consistent across all OSD groups, indicating minimal impact of ocular surface disease on accuracy. The coefficient of variation (CoV) for the trabecular iris-space area was about 20% for all cohorts. Ocular surface inflammation emerged as a key factor in dry eye, affecting immune-mediated and non-immune conditions alongside age-related ocular surface changes. While anterior chamber depth measurements showed variations, particularly in immune (CoV = 2.5%) and non-immune (CoV = 3.8%) OSD groups, parameters like anterior chamber width and angle to angle showed similar values among the cohorts. Keratometry measures remained stable despite OSD (CoV lower than 1%). Conclusion: The Tomey CASIA 2 demonstrated reliable repeatability for measuring anterior segment parameters in diverse OSD cohorts. Despite challenges posed by dry eye conditions, this technology holds promise in assessing OSD, suggesting potential clinical protocols similar to those in healthy controls.

Relative peripheral refraction in school children with different refractive errors using a novel multispectral refraction topographer.

AIM: To compare relative peripheral refraction (RPR) in Chinese school children with different refractive errors using multispectral refraction topography (MRT). METHODS: A total of 713 eyes of primary school children [172 emmetropia (E), 429 low myopia (LM), 80 moderate myopia (MM), and 32 low hypermetropia (LH)] aged 10 to 13y were analyzed. RPRs were measured using MRT without mydriasis. MRT results showed RPR at 0-15° (RPR 0-15), 15°-30° (RPR 15-30), and 30°-45° (RPR 30-45) annular in the inferior (RPR-I), superior (RPR-S), nasal (RPR-N), and temporal (RPR-T) quadrants. Spherical equivalent (SE) was detected and calculated using an autorefractor. RESULTS: There were significant differences of RPR 15-30 between groups MM [0.02 (-0.12; 0.18)] and LH [-0.13 (-0.36; 0.12)] (P<0.05), MM and E [-0.06 (-0.20; 0.10)] (P<0.05), and LM [-0.02 (-0.15; 0.15)] and E (P<0.05). There were also significant differences of RPR 30-45 between groups MM [0.45 (0.18; 0.74)] and E [0.29 (-0.09; 0.67)] (P<0.05), and LM [0.44 (0.14; 0.76)] and E (P<0.001). RPR values increased from the hyperopic to medium myopic group in each annular. There were significant differences of RPR-S between groups MM [-0.02 (-0.60; 0.30)] and E [-0.44 (-0.89; -0.04)] (P<0.001), and LM [-0.28 (-0.71; 0.12)] and E (P<0.05). There were also significant differences of RPR-T between groups MM [0.37 (0.21; 0.78)] and LH [0.14 (-0.52; 0.50)] (P<0.05), LM [0.41 (0.06; 0.84)] and LH (P<0.05), and LM and E [0.29 (-0.10; 0.68), P<0.05]. A Spearman's correlation analysis showed a negative correlation between RPR and SE in the 15°-30° (P=0.005), 30°-45° (P<0.05) annular (P=0.002), superior (P<0.001), and temporal (P=0.001) quadrants. CONCLUSION: Without pupil dilation, values for RPR 15-30, 30-45, RPR-S, and T shows significant differences between myopic eyes and emmetropia, and the differences are negatively correlated with SE.

Formation processes and mechanisms of a fault-controlled colluvial landslide in the Qinling-Daba Mountains, China.

Faults play a crucial role in shaping the formation and damage patterns of landslides in the mountainous region, particularly in Qinling-Daba (Qinba) area in China. On 6 October 2022, following a 4-day rainfall event totaling 221.5 mm, a landslide occurred in Hanwang Town, Shaanxi Province. The left boundary of the landslide coincided with a fault, which influence the formation and movement development of the landslide. To further understand and quantified the formation process and damage mechanism of the landslide, a comprehensive study was conducted, incorporating field investigations, local rainfall data, and various methods including unmanned aerial vehicles (UAVs), numerical simulations, and laboratory test. The results indicate that fault dictate the formation of the Lijiaping landslide by influencing the mechanical strength of the rock mass and the catchment topography in the landslide area. Due to fault, the rock mass in the landslide area is high fragmentation, with a softening coefficient of about 0.52. Weathering resulted in numerous residual and slope sediments in the landslide area, providing ample material for the landslide. Meanwhile, the fault activity led to a wedge-shaped topography in the landslide area, with an average Terrain Wetness Index (TWI) of 3.43, significantly higher than the Hanwang Township average of 1.47. This creates a hydrogeological structure favorable for landslides. Numerical simulations revealed that the maximum velocity of the landslide reached 5.05 m/s and the maximum displacement was 53.18 m, both occurring in the central part of the landslide. These findings offer crucial scientific insights for understanding and preventing similar geological hazards.

Super-Resolved Protein Imaging Using Bifunctional Light-Up Aptamers.

Efficient labeling methods for protein visualization with minimal tag size and appropriate photophysical properties are required for single-molecule localization microscopy (SMLM), providing insights into the organization and interactions of biomolecules in cells at the molecular level. Among the fluorescent light-up aptamers (FLAPs) originally developed for RNA imaging, RhoBAST stands out due to its remarkable brightness, photostability, fluorogenicity, and rapid exchange kinetics, enabling super-resolved imaging with high localization precision. Here, we expand the applicability of RhoBAST to protein imaging by fusing it to protein-binding aptamers. The versatility of such bifunctional aptamers is demonstrated by employing a variety of protein-binding aptamers and different FLAPs. Moreover, fusing RhoBAST with the GFP-binding aptamer AP3 facilitates high- and super-resolution imaging of GFP-tagged proteins, which is particularly valuable in view of the widespread availability of plasmids and stable cell lines expressing proteins fused to GFP. The bifunctional aptamers compare favorably with standard antibody-based immunofluorescence protocols, as they are 7-fold smaller than antibody conjugates and exhibit higher bleaching-resistance. We demonstrate the effectiveness of our approach in super-resolution microscopy in secondary mammalian cell lines and primary neurons by RhoBAST-PAINT, an SMLM protein imaging technique that leverages the transient binding of the fluorogenic rhodamine dye SpyRho to RhoBAST.

Biomimetic design and integrated biofabrication of an in-vitro three-dimensional multi-scale multilayer cortical model.

The lack of accurate and reliable in vitro brain models hinders the development of brain science and research on brain diseases. Owing to the complex structure of the brain tissue and its highly nonlinear characteristics, the construction of brain-like in vitro tissue models remains one of the most challenging research fields in the construction of living tissues. This study proposes a multi-scale design of a brain-like model with a biomimetic cortical structure, which includes the macroscopic structural features of six layers of different cellular components, as well as micrometer-scale continuous fiber structures running through all layers vertically. To achieve integrated biomanufacturing of such a complex multi-scale brain-like model, a multi-material composite printing/culturing integrated bioprinting platform was developed in-house by integrating cell-laden hydrogel ink direct writing printing and electrohydrodynamic fiber 3D printing technologies. Through integrated bioprinting, multi-scale models with different cellular components and fiber structural parameters were prepared to study the effects of macroscopic and microscopic structural features on the directionality of neural cells, as well as the interaction between glial cells and neurons within the tissue model in a three-dimensional manner. The results revealed that the manufactured in vitro biomimetic cortical model achieved morphological connections between the layers of neurons, reflecting the structure and cellular morphology of the natural cortex. Micrometer-scale (10 µm) cross-layer fibers effectively guided and controlled the extension length and direction of the neurites of surrounding neural cells but had no significant effect on the migration of neurons. In contrast, glial cells significantly promoted the migration of surrounding PC12 cells towards the glial layer but did not contribute to the extension of neurites. This study provides a basis for the design and manufacture of accurate brain-like models for the functionalization of neuronal tissues.

High Concentrate Supplementation during Late Pregnancy and Lambing Reduced Mortality of Triplet-Bearing Maternal Ewes.

Low survival of triplet-bearing ewes and their lambs represents lost production and a welfare issue. The effects of feed-on-offer (FOO; low: 1205 vs. high: 1980 kg DM/ha) and concentrate supplementation (low: 50 vs. high: 300+ g/ewe/day) levels during late pregnancy and lambing on the survival of triplet-bearing ewes and their lambs were investigated on 10 commercial farms using 1772 triplet-bearing Maternal ewes. Ewe and lamb survival were estimated at marking, and ewe body condition score (BCS) was recorded in late pregnancy and at marking. Although FOO treatment had no effect on triplet-bearing ewe mortality, receiving higher supplementation decreased mortality by 40% and increased BCS at marking by 0.14 compared with a lower supplementation (p < 0.05). Supplementation, FOO treatments, weather conditions during lambing and shelter availability had no effect on triplet-lamb survival. These findings suggest no additional benefit to triplet-bearing ewe survival when FOO levels exceed 1200 kg DM/ha during late pregnancy and lambing, but increased supplementation can reduce ewe mortality. Further research is required to determine the response to the supplementation level at lower FOO levels on triplet-bearing Merino ewes and their lambs and establish whether supplementation of triplet-bearing ewes during late pregnancy and lambing with higher levels of concentrates would be cost-effective.