Atto 465 Derivative Is a Nuclear Stain with Unique Excitation and Emission Spectra Useful for Multiplex Immunofluorescence Histochemistry

Multiplex immunofluorescence (mIF) is an effective technique for the maximal visualization of multiple target proteins in situ. This powerful tool is mainly limited by the spectral overlap of the currently available synthetic fluorescent dyes. The fluorescence excitation wavelengths ranging between 405 and 488 nm are rarely used in mIF imaging and serve as a logical additional slot for a fluorescent probe. In the present study, we demonstrate that the addition of 2,3,4,5,6-pentafluoroaniline to Atto 465 NHS ester, creating Atto 465-pentafluoroaniline (Atto 465-p), generates a bright nuclear stain in the violet-blue region of the visible spectrum. This allows the 405 nm excitation and emission, classically used for nuclear counterstains, to be used for the detection of another target protein. This increases the flexibility of the mIF panel and, with appropriate staining and microscopy, enables the quantitative analysis of at least six targets in one tissue section. (J Histochem Cytochem XX: XXX–XXX, XXXX)

CNN-Enhanced Multi-Indices Patch-Based Classification: A Case Study of Guwahati City

Land use and land cover (LULC) provides a way to classify objects on the surface of Earth. This paper aims to identify the varying land cover classes by stacking of 6 spectral bands and 10 different generated indices from those bands together. We have considered the multispectral images of Landsat 7 for our research. It is seen that instead of using only basic spectral bands (blue, green, red, nir, swir1 and swir2) for classification, stacking relevant indices of multiple target classes like ndvi, evi, nbr, BU, etc. with basic bands generates more precise results. In this study, we have used automated clustering techniques for generating 5 different class labels for training the model. These labels are further used to develop a predictive model to classify LULC classes. The proposed classifier is compared with the SVM and KNN classifiers. The results show that this proposed strategy gives preferable outcomes over other techniques. After training the model over 50 epochs, an accuracy of 93.29% is achieved. Keywords: Land use, land cover, CNN, ISODATA, indices

Sustainable development and health assessment model of higher education in India: A mathematical modeling approach

The Coronavirus Disease 2019 has resulted in a transition from physical education to online learning, leading to a collapse of the established educational order and a wisdom test for the education governance system. As a country seriously affected by the pandemic, the health of the Indian higher education system urgently requires assessment to achieve sustainable development and maximize educational externalities. This research systematically proposes a health assessment model from four perspectives, including educational volume, efficiency, equality, and sustainability, by employing the Technique for Order Preference by Similarity to an Ideal Solution Model, Principal Component Analysis, DEA-Tobit Model, and Augmented Solow Model. Empirical results demonstrate that India has high efficiency and an absolute health score in the higher education system through multiple comparisons between India and the other selected countries while having certain deficiencies in equality and sustainability. Additionally, single-target and multiple-target path are simultaneously proposed to enhance the Indian current education system. The multiple-target approach of the India-China-Japan-Europe-USA process is more feasible to achieve sustainable development, which would improve the overall health score from .351 to .716. This finding also reveals that the changes are relatively complex and would take 91.5 years considering the relationship between economic growth rates and crucial indicators. Four targeted policies are suggested for each catching-up period, including expanding and increasing the social funding sources, striving for government expenditure support to improve infrastructures, imposing gender equality in education, and accelerating the construction of high-quality teachers.

Evaluating complex interventions in international development

Complex interventions are those that are characterized by multiple components, multiple stakeholders, or multiple target populations. They may also be interventions that incorporate multiple processes of behavioral change. While such interventions are very common and receive a large proportion of development aid budgets, they are rarely subject to rigorous evaluations. The CEDIL Methods Working Paper, Evaluating Complex Interventions in International Development reviews promising methods for the evaluation of complex interventions that are new or have been used in a limited way. It offers a taxonomy of complex interventions in international development and draws on literature to discuss several methods that can be used to evaluate these interventions. The paper focuses its attention on methods that address causality and allow us to state conclusively whether an intervention works or not. It shows that several rigorous methods developed in different disciplines can be adapted and used to evaluate complex interventions in international development.

Evaluation of the impact of teaching on delineation variation during a virtual stereotactic ablative radiotherapy contouring workshop

Introduction: Variation in delineation of target volumes/organs at risk (OARs) is well recognised in radiotherapy and may be reduced by several methods including teaching. We evaluated the impact of teaching on contouring variation for thoracic/pelvic stereotactic ablative radiotherapy (SABR) during a virtual contouring workshop. Materials and methods: Target volume/OAR contours produced by workshop participants for three cases were evaluated against reference contours using DICE similarity coefficient (DSC) and line domain error (LDE) metrics. Pre- and post-workshop DSC results were compared using Wilcoxon signed ranks test to determine the impact of teaching during the workshop. Results: Of 50 workshop participants, paired pre- and post-workshop contours were available for 21 (42%), 20 (40%) and 22 (44%) participants for primary lung cancer, pelvic bone metastasis and pelvic node metastasis cases, respectively. Statistically significant improvements post-workshop in median DSC and LDE results were observed for 6 (50%) and 7 (58%) of 12 structures, respectively, although the magnitude of DSC/LDE improvement was modest in most cases. An increase in median DSC post-workshop ≥0·05 was only observed for GTVbone, IGTVlung and SacralPlex, and reduction in median LDE > 1 mm was only observed for GTVbone, CTVbone and SacralPlex. Post-workshop, median DSC values were >0·7 for 75% of structures. For 92% of the structures, post-workshop contours were considered to be acceptable or within acceptable variation following review by the workshop faculty. Conclusions: This study has demonstrated that virtual SABR contouring training is feasible and was associated with some improvements in contouring variation for multiple target volumes/OARs.

Detection of Target Sites Edited in Upland Cotton By The CRISPR/Cas9 System Mediated By Agrobacterium Rhizogenes

Background CRIPSR/Cas9 gene editing has the ability to effectively modify plant genomes. Multiple target sites usually were designed and the effective target sites were selected for editing. However, upland cotton is allotetraploid and is commonly considered as difficult and inefficient to transform. Therefore, it’s important to quickly identify feasibility of the target site. In this study, we use Agrobacterium rhizogenes K599 strain to infect cotton shoot meristem and induce them to grow hairy roots to detect the feasibility of a selected target designed in GhMYB25-like gene. Results We designed a sgRNA within the second exons of GhMYB25-likeA and GhMYB25-likeD and constructed the CRISPR vector. Transient hairy root transformation using A. rhizogenes K599 with four OD600s (0.4, 0.6,0.8, 1.0) was performed in Coker 312 (R15). The results show that A. rhizogenes at OD600 = 0.6–0.8 is the best concentration range for inducing cotton hairy roots. The other three cultivars (TM-1, Lumian 21, Zhongmian 49) were injected using A. rhizogenes K599 with OD600 = 0.6-0.8 and all produced hairy roots. We characterized ten R15 plants with hairy roots and detected different degrees of base deletions and insert at the target site in five R15 plants. Conclusion Overall, our data show A. rhizogenes-mediated transient hairy root transformation offers a rapid and efficient method to detect sgRNA feasibility in cotton.

Systematic end-to-end testing of multiple target treatments using the modular RUBY phantom

The RUBY head phantom in combination with the System QA insert MultiMet can be used for simultaneous point dose measurements at an isocentric and two off-axis positions. This study investigates the suitability of the system for systematic integral end-to-end testing of single-isocenter multiple target stereotactic treatments. Several volumetric modulated arc therapy plans were optimized on a planning CT of the phantom positioned in a stereotactic mask on the stereotactic treatment board. The plans were created for three artificial spherical target volumes centred around the measurement positions in the MultiMet insert. Target diameters between 5 and 40 mm were investigated. Coplanar and non-coplanar plans were optimized using the collapsed cone algorithm of the Oncentra Masterplan treatment planning system and recalculated with the Monte Carlo algorithm of the Monaco treatment planning system. Measurements were performed at an Elekta Synergy linear accelerator. The head phantom was positioned according to clinical workflow comprising immobilization and CBCT imaging. Simultaneous point dose measurements at all target positions were performed with three PinPoint 3D chambers (type 31022) as well as three microDiamond (type 60019) detectors and compared to the treatment planning system calculations. Furthermore, the angular dependence of the detector response was investigated to estimate the associated impact on the measured point dose values. Considering all investigated plans, PTV diameters and positions, the point doses calculated with the Monaco treatment planning system and the microDiamond measurements differed within 3.5%, whereas the PinPoint 3D showed differences of up to 6.9%. Point dose differences determined in comparison to the Oncentra Masterplan dose calculations were larger. The RUBY system was shown to be suitable for end-to-end testing of complex treatment scenarios such as single-isocenter multiple target plans.