Spectral absorption of visual pigments in stomatopod larval photoreceptors

Larval stomatopod eyes appear to be much simpler versions of adult compound eyes, lacking most of the visual pigment diversity and photoreceptor specializations. Our understanding of the visual pigment diversity of larval stomatopods, however, is based on four species, which severely limits our understanding of stomatopod eye ontogeny. To investigate several poorly understood aspects of stomatopod larval eye function, we tested two hypotheses surrounding the spectral absorption of larval visual pigments. First, we examined a broad range of species to determine if stomatopod larvae generally express a single, spectral class of photoreceptor. Using microspectrophotometry (MSP) on larvae captured in the field, we found data which further support this long-standing hypothesis. MSP was also used to test whether larval species from the same geographical region express visual pigments with similar absorption spectra. Interestingly, despite occupation of the same geographical location, we did not find evidence to support our second hypothesis. Rather, there was significant variation in visual pigment absorption spectra among sympatric species. These data are important to further our understanding of larval photoreceptor spectral diversity, which is beneficial to ongoing investigations into the ontogeny, physiology, and molecular evolution of stomatopod eyes. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s00359-015-1063-y) contains supplementary material, which is available to authorized users

Protocol for profiling in vitro intratumor heterogeneity using spatially annotated single-cell sequencing

Here, we present a protocol for spatially annotated single-cell sequencing, a technique for spatially profiling intratumor heterogeneity with deep single-cell RNA sequencing and single-cell resolution. By combining live-cell imaging and photopatterned illumination, we describe steps to identify regions of interest in an in vitro tumor model, label the selected cells with photoactivatable dyes, and isolate and subject them to scRNAseq. This protocol can be applied to a range of cell lines and could be expanded to tissue sections. For complete details on the use and execution of this protocol, please refer to Smit et al. (2022).1</p

Instant processing of large-scale image data with FACT, a real-time cell segmentation and tracking algorithm

Quantifying cellular characteristics from a large heterogeneous population is essential to identify rare, disease-driving cells. A recent development in the combination of high-throughput screening microscopy with single-cell profiling provides an unprecedented opportunity to decipher disease-driving phenotypes. Accurately and instantly processing large amounts of image data, however, remains a technical challenge when an analysis output is required minutes after data acquisition. Here, we present fast and accurate real-time cell tracking (FACT). FACT can segment ∼20,000 cells in an average of 2.5 s (1.9–93.5 times faster than the state of the art). It can export quantifiable features minutes after data acquisition (independent of the number of acquired image frames) with an average of 90%–96% precision. We apply FACT to identify directionally migrating glioblastoma cells with 96% precision and irregular cell lineages from a 24 h movie with an average F1 score of 0.91.</p

Instant processing of large-scale image data with FACT, a real-time cell segmentation and tracking algorithm

Quantifying cellular characteristics from a large heterogeneous population is essential to identify rare, disease-driving cells. A recent development in the combination of high-throughput screening microscopy with single-cell profiling provides an unprecedented opportunity to decipher disease-driving phenotypes. Accurately and instantly processing large amounts of image data, however, remains a technical challenge when an analysis output is required minutes after data acquisition. Here, we present fast and accurate real-time cell tracking (FACT). FACT can segment ∼20,000 cells in an average of 2.5 s (1.9–93.5 times faster than the state of the art). It can export quantifiable features minutes after data acquisition (independent of the number of acquired image frames) with an average of 90%–96% precision. We apply FACT to identify directionally migrating glioblastoma cells with 96% precision and irregular cell lineages from a 24 h movie with an average F1 score of 0.91.</p

Understanding the psychological mechanisms of return to sports readiness after anterior cruciate ligament reconstruction.

Purpose The psychological response to an Anterior Cruciate Ligament (ACL) injury is significant and can negatively impact return to sports outcomes. This study aimed to quantify the association between factors associated with return to sport using network analysis. Methods 441 participants who underwent primary ACL reconstruction. The 12-item ACL Return to Sport after Injury (ACL-RSI) scale was administered to all participants 12 months after surgery. Three network analyses were used to quantify the adjusted correlations between the 12 items of the ACL-RSI scale, and to determine the centrality indices of each item (i.e., the degree of connection with other items in the network). Further subgroup network analyses were conducted for those who had (n = 115) and had not returned (n = 326) to their pre-injury level of sport. Results The greatest adjusted correlation was between Q7 and Q9 (fear of re-injury and afraid of accidentally injuring knee) of the ACL-RSI (group 0.48 (95%CI [0.40 to 0.57])) across all three networks. The most important item in the network was Q12 (relaxed about sport) across all three networks. Individuals who did return to sport had greater Strength centrality for Q8 (confidence in knee, P = 0.014) compared to those who did not return to sport. Conclusion Fear of re-injury and being relaxed about playing sport were the two most important nodes in the network models that describe the return to sport readiness. The importance of knee confidence at influencing psychological readiness was greater in athletes who did return to sport compared to those who did not. Our findings provide candidate therapeutic targets that could inform future interventions designed to optimize return to sport rates in athletes post ACL reconstruction

Additions to Philippine Slender Skinks of the <i>Brachymeles bonitae </i>Complex (Reptilia: Squamata: Scincidae) III:a new species from Tablas Island

Davis, Drew R., Geheber, Aaron D., Watters, Jessa L., Penrod, Michelle L., Feller, Kathryn D., Ashford, Alissa, Kouri, Josh, Nguyen, Daniel, Shauberger, Kathryn, Sheatsley, Kyra, Winfrey, Claire, Wong, Rachel, Sanguila, Marites B., Brown, Rafe M., Siler, Cameron D. (2016): Additions to Philippine Slender Skinks of the Brachymeles bonitae Complex (Reptilia: Squamata: Scincidae) III: a new species from Tablas Island. Zootaxa 4132 (1), DOI: http://doi.org/10.11646/zootaxa.4132.1.

Additions to Philippine Slender Skinks of the <i>Brachymeles bonitae </i>Complex (Reptilia: Squamata: Scincidae) I:a new species from Lubang Island

Geheber, Aaron D., Davis, Drew R., Watters, Jessa L., Penrod, Michelle L., Feller, Kathryn D., Davey, Conner S., Ellsworth, Elyse D., Flanagan, Rachel L., Heitz, Brendan B., Moore, Tana, Nguyen, Marie D. C., Roberts, Austyn, Sutton, John, Sanguila, Marites B., Linkem, Charles W., Brown, Rafe M., Siler, Cameron D. (2016): Additions to Philippine Slender Skinks of the Brachymeles bonitae Complex (Reptilia: Squamata: Scincidae) I: a new species from Lubang Island. Zootaxa 4132 (1): 1-14, DOI: http://doi.org/10.11646/zootaxa.4132.1.

Genomic Exploration of Distinct Molecular Phenotypes Steering Temozolomide Resistance Development in Patient-Derived Glioblastoma Cells

Chemotherapy using temozolomide is the standard treatment for patients with glioblastoma. Despite treatment, prognosis is still poor largely due to the emergence of temozolomide resistance. This resistance is closely linked to the widely recognized inter- and intra-tumoral heterogeneity in glioblastoma, although the underlying mechanisms are not yet fully understood. To induce temozolomide resistance, we subjected 21 patient-derived glioblastoma cell cultures to Temozolomide treatment for a period of up to 90 days. Prior to treatment, the cells’ molecular characteristics were analyzed using bulk RNA sequencing. Additionally, we performed single-cell RNA sequencing on four of the cell cultures to track the evolution of temozolomide resistance. The induced temozolomide resistance was associated with two distinct phenotypic behaviors, classified as “adaptive” (ADA) or “non-adaptive” (N-ADA) to temozolomide. The ADA phenotype displayed neurodevelopmental and metabolic gene signatures, whereas the N-ADA phenotype expressed genes related to cell cycle regulation, DNA repair, and protein synthesis. Single-cell RNA sequencing revealed that in ADA cell cultures, one or more subpopulations emerged as dominant in the resistant samples, whereas N-ADA cell cultures remained relatively stable. The adaptability and heterogeneity of glioblastoma cells play pivotal roles in temozolomide treatment and contribute to the tumor’s ability to survive. Depending on the tumor’s adaptability potential, subpopulations with acquired resistance mechanisms may arise.</p