Principles of Spatial Transcriptomics Analysis: A Practical Walk-Through in Kidney Tissue

Spatial transcriptomic technologies capture genome-wide readouts across biological tissue space. Moreover, recent advances in this technology, including Slide-seqV2, have achieved spatial transcriptomic data collection at a near-single cell resolution. To-date, a repertoire of computational tools has been developed to discern cell type classes given the transcriptomic profiles of tissue coordinates. Upon applying these tools, we can explore the spatial patterns of distinct cell types and characterize how genes are spatially expressed within different cell type contexts. The kidney is one organ whose function relies upon spatially defined structures consisting of distinct cellular makeup. Thus, the application of Slide-seqV2 to kidney tissue has enabled us to elucidate spatially characteristic cellular and genetic profiles at a scale that remains largely unexplored. Here, we review spatial transcriptomic technologies, as well as computational approaches for cell type mapping and spatial cell type and transcriptomic characterizations. We take kidney tissue as an example to demonstrate how the technologies are applied, while considering the nuances of this architecturally complex tissue.

Pinch Anatomy of Forehead: An Injection Guide for Forehead Filler Treatment

With the popularity of filler injections globally, more and more injectors are using them for facial shaping and reversing the aging changes of the face. Detailed knowledge of facial anatomy, especially of important vessels and tissue planes is essential for injectors. While performing filler injections, injectors tend to pinch the tissue layers with their non-dominant hand for ease of doing the procedure. Such deformational forces cause some changes in the anatomy of tissue layers held in a pinch. During a deep pinch, the important arteries in that area can get pulled up in the pinch's tissue layers, or they can stay in their position being unaffected by pinching. The ‘pinch and pull’ can also improve the tissue space for injections, by pulling away the mobile tissue layers from the fixed ones. Knowledge of the ‘pinch anatomy’ in the forehead can be used to the injector's advantage to avoid important arteries and place filler in the correct plane. By knowing the anatomical changes during the pinched state of tissue layers, filler injections in the forehead can be performed with relative safety in the correct tissue plane.

CLINICALLY EVALUATED ACCURACY OF SINGLE GUIDED IMPLANTATIONS WITH TWO DIFFERENT SYSTEMS – PRELIMINARY RESULTS

Introduction: Guided single implant placement ensures an optimal implant position, enables in-surgery immediate loading, and creates a predisposition for predictable treatment results. Aim: This article aims to present the surgical and loading protocols and assess the accuracy of two guided implant systems. Materials and methods: Ten patients missing a single molar were included. The semi-guided procedures with the two implant systems – AB Dental and Alpha Bio-Tec, were randomly assigned to each participant. The computer-assisted planning was performed with Implant Studio (3Shape, Denmark). Surgical guides and long-term, temporary, screw-retained restorations were virtually generated within the 3Shape system. Surgical appliances were printed from SG resin (Formlabs, USA), and crowns were milled from Telio CAD (Ivoclar Vivadent, Lichtenstein). Planning accuracy was evaluated based on the crown’s fit and relation to the adjacent teeth and antagonists. Results and discussion: The patients included for preliminary analysis (6 female and 4 male) were with a mean age of 33.42. The youngest was 24, and the oldest 49 years old. There were no surgical and technical complications for the period of evaluation, which ranged from 1 to 3 months. The crown’s fit was excellent in 8 cases. In two cases, minor adjustments were necessary – 1 in relation to the antagonists and 1 to the adjacent teeth. In one case, a healing screw was placed for 48 hours to facilitate adequate soft-tissue space for the Ti-base. Conclusion: The preliminary results presented in this report suggest that the clinical and laboratory protocols used for guided implantation and immediate loading are highly efficient.

Optogenetic perturbations of RNA expression in tissue space

Quantifying gene expression in space, for example by spatial transcriptomics, is essential for describing the biology of cells and their interactions in complex tissues. Perturbation experiments, at single-cell resolution and conditional on both space and time, are necessary for dissecting the molecular mechanisms of these interactions. To this aim, we combined optogenetics and CRISPR technologies to activate or knock-down RNA of target genes, at single-cell resolution and in programmable spatial patterns. As a proof of principle, we optogenetically induced Sonic Hedgehog (SHH) signaling at a distinct spatial location within human neural organoids. This robustly induced known SHH spatial domains of gene expression – cell-autonomously and across the entire organoid. In principle, our approach can be used to induce or knock down RNAs from any gene of interest in specific spatial locations or patterns of complex biological systems.

Mobility and Urban Centralities: An Analysis Based on the Motorized Flows Attraction in Belo Horizonte/State of Minas Gerais/Brazil

The issues encompassing planning and management of the urban space are directly related to the transport system, of which the flow plays a central role. Therefore, an interdependence among urban tissue, space structuring, and demand for mobility are taken into account. The objective of this work, using the data taken from the last two Origin and Destination surveys (2002 and 2012), is to assess the possible reduced power of attraction of the Central Area of Belo Horizonte by means of the motorized flows as a result of the process of induction of power of attraction of new urban centralities through planning and management policies. The results presented in this work generally indicate that the spatial structure of Belo Horizonte has remained concentrated in the Central Area, though new flows seem to converge to the consolidation of new municipal centralities. Even if additional care in the interpretation of results is necessary due to the limited time cuts and the database used, besides the specific features of Belo Horizonte, the empirical evidence translated in the proposed indicators allow a more accurate reflection on the recent urban space configuration, with the connections established through the space mobility of the population.

Features of acute purulent lymphadenites and maxillofacial adenophlegmons course in children

Objective. The purpose of the research was to study the features of the course of acute purulent lymphadenites and adenophlegmons of the maxillofacial region in children, to describe the species composition of pathogens, to determine their sensitivity to antibiotics of various groups. Materials and methods. The medical histories of 147 patients were analyzed; microbiological studies were carried out (microscopic examination of clinical specimens with gram stain, cultural method, determination of sensitivity of isolated pathogen to antimicrobial drugs). Results. Most often, acute purulent lymphadenites and adenophlegmons were observed in the submandibular tissue space (40.1 %) and in the neck (38.1 %). The main causative agents of the inflammatory process were Staphylococcus aureus (38.1 %) and Streptococcus agalactiae (BHSA) (14.3 %). Staphylococcus aureus strains had the highest sensitivity to beta-lactam antibiotics in 83 %, to macrolides in 60.4 %, to fluoroquinolones in 56.6 %; 3.8 % showed resistance to lincomycin, 1.9 % to beta-lactams. The strains of Streptococcus agalactiae (BHSA) were 100 % sensitive to beta-lactams, 73.7 % to macrolides, they were resistant to lincomycin and macrolides (10.5 % each). Conclusions. The data obtained are of great practical importance, since they allow to correctly orient doctors in the choice of rational antibiotic therapy.

INFLAMMATION OF INTERSTITIAL CONNECTIVE TISSUE SPACE INDUCED BY LEAD ACETATE AND AMELIORATIVE AFTERMATH OF FICUSCARICAON RAT TESTIS

Objective: To observe inflammation of interstitial connective tissue space caused by Lead acetate in rat testis and ameliorative after math caused by Ficuscarica. Study Design: Laboratory based experimental study. Place and Duration of Study: Anatomy Department, Army Medical College Rawalpindi together with NIH (National Institute of Health) Islamabad, from Mar to Nov 2017. Methodology: Sprague Dawley male rats, 30 in quantity were chosen and 10 animals each partited into 3 groups. Treatments were given for 8 weeks, once daily. Group A was control group. Group B was treated with dosage of 30 mg/kg of Lead acetate. Group C was given dosage of 30 mg/kg of Lead acetate as well as 80 mg/kg of Ficuscarica. Twenty four hours after the concluding dose, animals were vivisected. For histological study, testis were fixed and stained with Haematoxylin and eosin. Interstitial connective tissue space thickness was morphometrically and assessed by SPSS version 22. A p-value <0.05 was considered significant in results. Results: Interstitial space thickness was significantly increased due to inflammation (>3 times normal) in group B in comparison to groups A and C. Thickness of space was slightly increased (<2 times normal) in group C in comparison to groups A due to reduction in inflammation. Conclusion: There was increased thickness of interstitium due to inflammation, cellular congestion and lymphocytic infiltration in rat’s testis because of lead acetate but concomitant dose of Ficuscarica protects against inflammation, venous congestion of interstitial space.

Bioengineered in vitro models of leukocyte–vascular interactions

Leukocytes continuously circulate our body through the blood and lymphatic vessels. To survey invaders or abnormalities and defend our body against them, blood-circulating leukocytes migrate from the blood vessels into the interstitial tissue space (leukocyte extravasation) and exit the interstitial tissue space through draining lymphatic vessels (leukocyte intravasation). In the process of leukocyte trafficking, leukocytes recognize and respond to multiple biophysical and biochemical cues in these vascular microenvironments to determine adequate migration and adhesion pathways. As leukocyte trafficking is an essential part of the immune system and is involved in numerous immune diseases and related immunotherapies, researchers have attempted to identify the key biophysical and biochemical factors that might be responsible for leukocyte migration, adhesion, and trafficking. Although intravital live imaging of in vivo animal models has been remarkably advanced and utilized, bioengineered in vitro models that recapitulate complicated in vivo vascular structure and microenvironments are needed to better understand leukocyte trafficking since these in vitro models better allow for spatiotemporal analyses of leukocyte behaviors, decoupling of interdependent biological factors, better controlling of experimental parameters, reproducible experiments, and quantitative cellular analyses. This review discusses bioengineered in vitro model systems that are developed to study leukocyte interactions with complex microenvironments of blood and lymphatic vessels. This review focuses on the emerging concepts and methods in generating relevant biophysical and biochemical cues. Finally, the review concludes with expert perspectives on the future research directions for investigating leukocyte and vascular biology using the in vitro models.