Downregulation of Fidgetin-Like 2 Increases Microglial Function: The Relationship Between Microtubules, Morphology, and Activity.

The microtubule cytoskeleton regulates microglial morphology, motility, and effector functions. The microtubule-severing enzyme, fidgetin-like 2 (FL2), negatively regulates cell motility and nerve regeneration, making it a promising therapeutic target for central nervous system injury. Microglia perform important functions in response to inflammation and injury, but how FL2 affects microglia is unclear. In this study, we investigated the role of FL2 in microglial morphology and injury responses in vitro. We first determined that the pro-inflammatory stimulus, lipopolysaccharide (LPS), induced a dose- and time-dependent reduction in FL2 expression associated with reduced microglial ramification. We then administered nanoparticle-encapuslated FL2 siRNA to knockdown FL2 and assess microglial functions compared to negative control siRNA and vehicle controls. Time-lapse live-cell microscopy showed that FL2 knockdown increased the velocity of microglial motility. After incubation with fluorescently labeled IgG-opsonized beads, FL2 knockdown increased phagocytosis. Microglia were exposed to low-dose LPS after nanoparticle treatment to model injury-induced cytokine secretion. FL2 knockdown enhanced LPS-induced cytokine secretion of IL-1α, IL-1ß, and TNFα. These results identify FL2 as a regulator of microglial morphology and suggest that FL2 can be targeted to increase or accelerate microglial injury responses.

Propagating pluripotency - The conundrum of self-renewal.

The discovery of mouse embryonic stem cells in 1981 transformed research in mammalian developmental biology and functional genomics. The subsequent generation of human pluripotent stem cells (PSCs) and the development of molecular reprogramming have opened unheralded avenues for drug discovery and cell replacement therapy. Here, I review the history of PSCs from the perspective that long-term self-renewal is a product of the in vitro signaling environment, rather than an intrinsic feature of embryos. I discuss the relationship between pluripotent states captured in vitro to stages of epiblast in the embryo and suggest key considerations for evaluation of PSCs. A remaining fundamental challenge is to determine whether naïve pluripotency can be propagated from the broad range of mammals by exploiting common principles in gene regulatory architecture.

Structural Elucidation and Absolute Stereochemistry for Pharma Compounds Using MicroED.

Microcrystal electron diffraction (microED) is an emerging technique for rapid crystallographic analysis of small molecule micro- and nanocrystals. In this report, we evaluate the applicability of microED to pharmaceutical compounds through the analysis of 30 samples obtained from the process and medicinal chemistry groups at Amgen Inc. Using only 40 h of microscope time, 15 of 30 crystal structures were elucidated. From these crystal structures, all chiral compounds had the correct absolute stereochemistry assigned by dynamical refinement of continuous rotation electron diffraction data, confirming dynamical refinement as a promising tool for the absolute stereochemistry determination of pharmaceutically relevant compounds.

Alpha-Gal Syndrome and Cardiac Surgery.

Galactose-alpha-1,3-galactose (alpha-gal) is a carbohydrate expressed by all mammals except for humans and certain old-world primates. It can be found in a plethora of products derived from mammals, including milk, organs, skeletal muscle and gelatin, in addition to products prepared with mammalian cells or constituents. In the late 2000s, an association between tick bites and the development of immunoglobulin E antibodies to the alpha-gal carbohydrate was discovered. The term "alpha-gal syndrome" (AGS) was then coined to describe allergic reactions to mammalian meat or other alpha-gal-containing products derived from mammals. Symptoms are often delayed several hours from consumption and can be urticarial and/or gastrointestinal. Medications and bioprosthetic inserts derived from mammals were also noted to cause allergic reactions in affected patients. Cardiac surgery, in particular, is considered high risk, given that unfractionated heparin has a bovine or porcine origin and is administered in large doses for cardiopulmonary bypass. Bioprosthetic valves have similar origins and risks. Awareness of AGS in cardiac surgery patients can lead to decreased risk preoperatively and inform management perioperatively and postoperatively. In this narrative review, we have reviewed the published literature relevant to AGS in patients undergoing cardiac surgery and shared our treatment approach.

Multi-scale signaling and tumor evolution in high-grade gliomas.

Although genomic anomalies in glioblastoma (GBM) have been well studied for over a decade, its 5-year survival rate remains lower than 5%. We seek to expand the molecular landscape of high-grade glioma, composed of IDH-wildtype GBM and IDH-mutant grade 4 astrocytoma, by integrating proteomic, metabolomic, lipidomic, and post-translational modifications (PTMs) with genomic and transcriptomic measurements to uncover multi-scale regulatory interactions governing tumor development and evolution. Applying 14 proteogenomic and metabolomic platforms to 228 tumors (212 GBM and 16 grade 4 IDH-mutant astrocytoma), including 28 at recurrence, plus 18 normal brain samples and 14 brain metastases as comparators, reveals heterogeneous upstream alterations converging on common downstream events at the proteomic and metabolomic levels and changes in protein-protein interactions and glycosylation site occupancy at recurrence. Recurrent genetic alterations and phosphorylation events on PTPN11 map to important regulatory domains in three dimensions, suggesting a central role for PTPN11 signaling across high-grade gliomas.

"Shoe Shop" Lacing Technique: A New Biceps-Augmented Knotless Suture for Arthroscopic Rotator Cuff Repair.

Advancements in rotator cuff tear repair have led to innovative techniques for complex cases. In this article, we introduce the "shoe shop" lacing technique, a knotless, side-to-side, and tendon-to-bone suture method augmented with the long head of the biceps tendon (LHBT) for anterior margin-deficient massive rotator cuff tears. This approach offers simplicity, durability, and potential advantages in biomechanics. The LHBT integration and knotless sutures make it a promising solution for challenging tear patterns. This technique provides an attractive option for arthroscopic repair, improving outcomes in cases where anterior cable reinforcement is essential.

ERK signalling eliminates Nanog and maintains Oct4 to drive the formative pluripotency transition.

Naïve epiblast cells in the embryo and pluripotent stem cells in vitro undergo developmental progression to a formative state competent for lineage specification. During this transition, transcription factors and chromatin are rewired to encode new functional features. Here, we examine the role of mitogen-activated protein kinase (ERK1/2) signalling in pluripotent state transition. We show that a primary consequence of ERK activation in mouse embryonic stem cells is elimination of Nanog, which precipitates breakdown of the naïve state gene regulatory network. Variability in pERK dynamics results in heterogeneous loss of Nanog and metachronous state transition. Knockdown of Nanog allows exit without ERK activation. However, transition to formative pluripotency does not proceed and cells collapse to an indeterminate identity. This outcome is due to failure to maintain expression of the central pluripotency factor Oct4. Thus, during formative transition ERK signalling both dismantles the naïve state and preserves pluripotency. These results illustrate how a single signalling pathway can both initiate and secure transition between cell states.

Latarjet procedure restores range of motion at 6 months postoperatively: a prospective cohort study using motion capture analysis.

BACKGROUND: There is a common concern that range of motion (ROM) is negatively affected by the Latarjet procedure. We hypothesize that the Latarjet procedure results in full recovery of ROM postoperatively and significantly improved patient-reported outcome measures. METHODS: Patient data were prospectively collected from a randomized controlled trial to analyze outcomes after open Latarjet procedure. Inclusion criteria involved a minimum follow-up of 6 months and unilateral shoulder instability. Study outcome was assessed by postoperative ROM at 6 months postoperatively and compared to the preoperative ROM of the ipsilateral shoulder as well as the ROM of the unaffected contralateral shoulder. All ROM measurements were performed using a motion capture system to ensure consistent and reliable measurements. RESULTS: The study included a total of 84 patients. ROM was measured in external rotation with the shoulder adducted (ER1), external rotation with the shoulder abducted 90° (ER2), internal rotation with the arm abducted 90° (IR2), and active forward elevation (AE). The mean difference in ROM between the operated arm vs. the contralateral healthy arm at 6 months postoperatively was 3.4° in ER1 (P = .19), 4.2° in ER2 (P = .086), 2.2° in IR2 (P = .36), and 2.4° in AE (P = .045). Subanalysis of patients with and without sling use revealed no significant difference in ROM between the operated shoulder and the contralateral shoulder at 6 months in either group, with the exception of ER2 in the sling group. In this latter group, ROM was 71° in the operated arm and 79° in the contralateral arm (P = .0094). Mean preoperative pain score was 25.7 (95% CI 21.4-30.1) vs. 13.0 postoperatively at 6 months (95% CI 9.50-16.5) (P < .00001). Mean preoperative Single Assessment Numeric Evaluation instability was 42.9 (95% CI 38.4-47.3) vs. 86.2 postoperatively at 6 months (95% CI 83.6-88.7) (P < .00001). Mean preoperative Rowe score was 38.5 (95% CI 34.3-42.7) vs. 84.3 at postoperative month 6 (95% CI 81.1-87.4) (P < .00001). CONCLUSIONS: Latarjet procedure performed for anterior instability using a capsular repair results in complete ROM recovery in ER1, ER2, and IR2 at 6 months postoperatively, with only a slight discrepancy in active elevation. Sling use after the Latarjet procedure results in no benefit over postoperative recovery without the use of a sling. Sling use negatively affects the ROM in ER2, taking as reference the contralateral arm of the same patient, when compared to patients who did not use a sling postoperatively.

Branching topology of the human embryo transcriptome revealed by Entropy Sort Feature Weighting.

Analysis of single cell transcriptomics (scRNA-seq) data is typically performed after subsetting to highly variable genes (HVGs). Here, we show that Entropy Sorting provides an alternative mathematical framework for feature selection. On synthetic datasets, continuous Entropy Sort Feature Weighting (cESFW) outperforms HVG selection in distinguishing cell-state-specific genes. We apply cESFW to six merged scRNA-seq datasets spanning human early embryo development. Without smoothing or augmenting the raw counts matrices, cESFW generates a high-resolution embedding displaying coherent developmental progression from eight-cell to post-implantation stages and delineating 15 distinct cell states. The embedding highlights sequential lineage decisions during blastocyst development, while unsupervised clustering identifies branch point populations obscured in previous analyses. The first branching region, where morula cells become specified for inner cell mass or trophectoderm, includes cells previously asserted to lack a developmental trajectory. We quantify the relatedness of different pluripotent stem cell cultures to distinct embryo cell types and identify marker genes of naïve and primed pluripotency. Finally, by revealing genes with dynamic lineage-specific expression, we provide markers for staging progression from morula to blastocyst.

A feasibility cadaver study for placing screws in various pelvic osseous fracture pathways using a robotic arm.

INTRODUCTION: The use of a robotic system for the placement of pedicle screws in spine surgeries is well documented in the literature. However, there is only a single report in the United States describing the use of a robotic system to place two screws in osseous fixation pathways (OFPs) commonly used in the treatment of pelvic and acetabular fractures in a simulated bone model. The purpose of this study was to demonstrate the use of a robotic system to place screws in multiple, clinically relevant OFPs in a cadaveric model and to quantitatively measure accuracy of screw placement relative to the preoperative plan. METHODS: A single cadaveric specimen was obtained for the purpose of this study. All surrounding soft tissues were left intact. Screws were placed in OFPs, namely iliosacral (IS), trans-sacral (TS), Lateral Compression-II (LC-II), antegrade anterior column (AC) and antegrade posterior column (PC) of the right hemipelvis using standard, fluoroscopically assisted percutaneous or mini-open technique. Following the placement of screws into the right hemipelvis using standard techniques, screws were planned and placed in the same OFPs of the contralateral hemipelvis using the commercially available ExcelsiusGPS® robotic system (Globus Medical Inc., Audubon, PA). After robotic-assisted screw placement, a post-procedure CT scan was obtained to evaluate actual screw placement against the pre-procedure plan. A custom-made image analysis program was devised to measure screw tip/tail offset and angular offset on axial and sagittal planes. RESULTS: For different OFPs, the mean tip offset, tail offset and angular offsets were 1.6 ± 0.9 mm (Range 0.0-3.6 mm), 1.4 ± 0.4 mm (Range 0.3-2.5 mm) and 1.1 ± 0.4° (Range 0.5-2.1), respectively. CONCLUSION: In this feasibility study, surgeons were able to place screws into the clinically relevant fracture pathways of the pelvis using ExcelsiusGPS® for robotic-assisted surgery. The measured accuracy was encouraging; however, further investigation is needed to demonstrate that robotic-assisted surgery can be used to successfully place the screws in the bony corridors of the pelvis to treat traumatic pelvic injuries.

Insuficiente aplicación de Medicina Natural y Tradicional en afecciones relacionadas con la ortopedia y la traumatología

La Medicina Natural y Tradicional aplica sus diferentes modalidades en la práctica médica moderna, segura, económica, inocua y simple; y con pocos efectos colaterales resuelve grandes problemas de salud en situaciones especiales o de desastres con recursos limitados. De ahí que el egresado de programas de especialización de las ciencias médicas cuenta con sólidos conocimientos para su aplicación, especialmente en las afecciones osteomioarticulares, donde la especialidad de ortopedia y traumatología tiene un papel primordial para identificar las causas y consecuencias de la insuficiencia de su aplicación en pacientes portadores de afecciones relacionadas con esta especialidad. A través de un estudio crítico reflexivo basado en el análisis y la síntesis, así como de la constatación de la validez de los puntos de vista asumidos como referentes en el proceso de superación del especialista de ortopedia y traumatología, se pudo mostrar cómo se ha concebido la superación profesional en el Hospital General Dr. Juan Bruno Zayas Alfonso, al integrar adecuadamente los conocimientos y las competencias. Esto se expresó en la elaboración e implementación de una estrategia de superación profesional, que permitió la actualización de relevantes conocimientos necesarios en el campo de la MNT para el desempeño de este profesional. Se concluyó que era insuficiente el nivel de conocimiento de los contenidos, como también los cursos ofertados para su desarrollo, lo que trajo como consecuencia una atención médica sin calidad.

Natural and traditional medicine applies its different modalities in modern, safe, economical, innocuous and simple medical practice; with few collateral effects, it solves major health problems in special or disaster situations with limited resources. Hence, the graduate of specialization programs in medical sciences possesses solid knowledge for its application, especially in osteomyoarticular conditions, in which the specialty of orthopedics and traumatology has a key role in identifying the causes and consequences of its unsatisfactory application in patients with conditions related to this specialty. Through a reflexive and critical study based on analysis and synthesis, as well as the verification of the validity of the viewpoints assumed as referents in the process of improvement of the orthopedics and traumatology specialist, it was possible to show how professional improvement has been conceived at Hospital General Dr. Juan Bruno Zayas Alfonso, by adequately integrating knowledge and competences. This was expressed through the elaboration and implementation of a professional improvement strategy, which allowed updating relevant knowledge necessary in the field of natural and traditional medicine for the performance of this professional. The level of knowledge about the contents was concluded to be insufficient, while the courses offered for its development were so, which resulted in a medical care without quality.

Estrategia de superación profesional para especialistas de Ortopedia y Traumatología en Medicina Natural y Tradicional

Introducción: La Medicina Natural y Tradicional (MNT) tiene como objetivo integrar conocimientos de sus modalidades a la práctica médica, a partir de una adecuada selección y fundamentación científica. Desde su desarrollo en el proceso de superación profesional en especialidades como la Ortopedia y Traumatología logra atención médica de calidad, se ajusta a las condiciones sociohistóricas actuales y al desarrollo de nuevos estilos de actuación, mejora las relaciones interpersonales y fomenta los valores ético-morales en íntima interrelación con la comunidad. Objetivo: Exponer fundamentos que sustentan el diseño y la aplicación de una estrategia de superación profesional en especialistas de Ortopedia y Traumatología para la aplicación de la Medicina Natural y Tradicional. Métodos: Se realizó un estudio pedagógico, prospectivo, que utilizó métodos teóricos y empíricos. Para la elaboración de la Estrategia de Superación Profesional se utilizó la modelación, que posibilitó el proceso de concreción-abstracción con sus interrelaciones y vínculos con la aplicación de la Medicina Natural y Tradicional. Se asumieron los criterios utilizados por la Dra. C. Yerina Figueredo. La población fue de 43 especialistas de Ortopedia y Traumatología del Hospital "Juan Bruno Zayas Alfonso", de Santiago de Cuba, entre julio y diciembre de 2021. Resultados: Se demostró alto nivel de satisfacción con la aplicación de la estrategia y la consulta de expertos evidenció mejora en el desempeño de estos profesionales. Conclusiones: La actualización en las modalidades de la Medicina Natural y Tradicional en la superación de los Especialistas de Ortopedia y Traumatología aportó un grupo de regularidades a la formación permanente y continuada, que enriquecen las ciencias de la educación médica.

Introduction: Natural and traditional medicine aims at integrating knowledge within its modalities to medical practice, based on an adequate selection and scientific foundation. Since its development as part of the professional improvement process in specialties such as Orthopedics and Traumatology, it achieves quality medical care, adjusts itself to the current sociohistorical conditions and the development of new action styles, improves interpersonal relationships, as well as promotes ethical and moral values in close relationship with the community. Objective: To present the foundations that support the design and application of a professional improvement strategy with Orthopedics and Traumatology specialists for the application of natural and traditional medicine. Methods: A pedagogical and prospective study was carried out, using theoretical and empirical methods. Modeling was used for the elaboration of the professional improvement strategy, which made possible the concretion-abstraction process with its interrelations and links in view of the application of natural and traditional medicine. The criteria used by Yerina Figueredo, Ph.D., were assumed. The population was 43 Orthopedics and Traumatology specialists from Hospital Juan Bruno Zayas Alfonso, of Santiago de Cuba, between July and December 2021. Results: A high level of satisfaction with the application of the strategy was demonstrated and the consultation with experts evidenced an improvement in the performance of these professionals. Conclusions: The updating in the modalities of natural and traditional medicine for the training of Orthopedics and Traumatology specialists contributed a group of regularities to permanent and continuous training, which enriches the sciences of medical education.

Epigenetic regulation during cancer transitions across 11 tumour types.

Chromatin accessibility is essential in regulating gene expression and cellular identity, and alterations in accessibility have been implicated in driving cancer initiation, progression and metastasis1-4. Although the genetic contributions to oncogenic transitions have been investigated, epigenetic drivers remain less understood. Here we constructed a pan-cancer epigenetic and transcriptomic atlas using single-nucleus chromatin accessibility data (using single-nucleus assay for transposase-accessible chromatin) from 225 samples and matched single-cell or single-nucleus RNA-sequencing expression data from 206 samples. With over 1 million cells from each platform analysed through the enrichment of accessible chromatin regions, transcription factor motifs and regulons, we identified epigenetic drivers associated with cancer transitions. Some epigenetic drivers appeared in multiple cancers (for example, regulatory regions of ABCC1 and VEGFA; GATA6 and FOX-family motifs), whereas others were cancer specific (for example, regulatory regions of FGF19, ASAP2 and EN1, and the PBX3 motif). Among epigenetically altered pathways, TP53, hypoxia and TNF signalling were linked to cancer initiation, whereas oestrogen response, epithelial-mesenchymal transition and apical junction were tied to metastatic transition. Furthermore, we revealed a marked correlation between enhancer accessibility and gene expression and uncovered cooperation between epigenetic and genetic drivers. This atlas provides a foundation for further investigation of epigenetic dynamics in cancer transitions.

Differential chromatin accessibility and transcriptional dynamics define breast cancer subtypes and their lineages.

Breast cancer is a heterogeneous disease, and treatment is guided by biomarker profiles representing distinct molecular subtypes. Breast cancer arises from the breast ductal epithelium, and experimental data suggests breast cancer subtypes have different cells of origin within that lineage. The precise cells of origin for each subtype and the transcriptional networks that characterize these tumor-normal lineages are not established. In this work, we applied bulk, single-cell (sc), and single-nucleus (sn) multi-omic techniques as well as spatial transcriptomics and multiplex imaging on 61 samples from 37 breast cancer patients to show characteristic links in gene expression and chromatin accessibility between breast cancer subtypes and their putative cells of origin. We applied the PAM50 subtyping algorithm in tandem with bulk RNA-seq and snRNA-seq to reliably subtype even low-purity tumor samples and confirm promoter accessibility using snATAC. Trajectory analysis of chromatin accessibility and differentially accessible motifs clearly connected progenitor populations with breast cancer subtypes supporting the cell of origin for basal-like and luminal A and B tumors. Regulatory network analysis of transcription factors underscored the importance of BHLHE40 in luminal breast cancer and luminal mature cells, and KLF5 in basal-like tumors and luminal progenitor cells. Furthermore, we identify key genes defining the basal-like ( PRKCA , SOX6 , RGS6 , KCNQ3 ) and luminal A/B ( FAM155A , LRP1B ) lineages, with expression in both precursor and cancer cells and further upregulation in tumors. Exhausted CTLA4-expressing CD8+ T cells were enriched in basal-like breast cancer, suggesting altered means of immune dysfunction among breast cancer subtypes. We used spatial transcriptomics and multiplex imaging to provide spatial detail for key markers of benign and malignant cell types and immune cell colocation. These findings demonstrate analysis of paired transcription and chromatin accessibility at the single cell level is a powerful tool for investigating breast cancer lineage development and highlight transcriptional networks that define basal and luminal breast cancer lineages.

Glenoid-based reference system to differentiate shoulder pathologies on plain radiographs.

BACKGROUND: Shoulder radiographs are used for evaluation and the planning of treatment of various pathologies. Making a diagnosis of these pathologies on plain radiographs occurs by recognizing the relationship of the humeral head on the registry of the glenoid. Quantification of these changes in registry does not currently exist. We hypothesize that a geometric relationship of the humeral head and the glenoid exists that is defined on an anteroposterior Grashey view radiograph by the relationship of the best-fit circle of the humeral head relative to the best-fit circle of the glenoid such that relative measurements will define the normal shoulder and the pathologic shoulder. METHODS: One hundred fifty-six shoulders were included: 53 normal shoulders, 51 with primary glenohumeral osteoarthritis (GHOA), and 52 with cuff tear arthropathy (CTA). Humeral head best-fit circle was used to define the circle of the humeral head (cHH). A glenoid best-fit circle (cG) was defined by the following rules: (1) best fit of the glenoid articular surface and (2) was limited by the acromion such that either (a) it reaches maximal interaction with the inferior surface of the acromion or (b) the perimeter of the circle is at the lateralmost point of the acromion. The relationship between cHH and cG is defined by measurement of cHH in horizontal and vertical planes relative to the glenoid circle reference. The horizontal displacement angle (HDA) measures the horizontal position of cHH relative to cG, representing the degree of medialization toward the glenoid. The vertical displacement angle (VDA) measures the vertical position of cHH relative to cG, representing the degree of superiorization toward the acromion. Angles were compared by diagnosis and sex. RESULTS: The mean HDA was 61.0° (95% confidence interval [CI] 60.3°-61.7°) in normal shoulders, 79.9° (95% CI 76.9°-82.9°) in GHOA, and 63.4° (95% CI 61.7°-65.1°) in CTA (P < .001). The mean VDA was 43.1° (95% CI 42.2°-44.0°) in normal shoulders, 40.9° (95% CI 39.9°-42.0°) in GHOA, and 59.7° (95% CI 57.6°-61.7°) in CTA (P < .001). Interobserver reliability was 0.991 (95% CI 0.94-1.0) and intraobserver reliability was 0.998 (95% CI 0.99-1.0). The geometric relationship of cHH to the glenoid circle reference was plotted for each group. CONCLUSION: A geometric relationship exists of the humeral head in reference to the glenoid circle. Together, the HDA and the VDA distinguish between a normal shoulder and those with GHOA or CTA. This suggests that this novel methodology may provide a preoperative planning tool that is easily accessible.

Epigenetic dynamics during capacitation of naïve human pluripotent stem cells.

Human pluripotent stem cells (hPSCs) are of fundamental relevance in regenerative medicine. Naïve hPSCs hold promise to overcome some of the limitations of conventional (primed) hPSCs, including recurrent epigenetic anomalies. Naïve-to-primed transition (capacitation) follows transcriptional dynamics of human embryonic epiblast and is necessary for somatic differentiation from naïve hPSCs. We found that capacitated hPSCs are transcriptionally closer to postimplantation epiblast than conventional hPSCs. This prompted us to comprehensively study epigenetic and related transcriptional changes during capacitation. Our results show that CpG islands, gene regulatory elements, and retrotransposons are hotspots of epigenetic dynamics during capacitation and indicate possible distinct roles of specific epigenetic modifications in gene expression control between naïve and primed hPSCs. Unexpectedly, PRC2 activity appeared to be dispensable for the capacitation. We find that capacitated hPSCs acquire an epigenetic state similar to conventional hPSCs. Significantly, however, the X chromosome erosion frequently observed in conventional female hPSCs is reversed by resetting and subsequent capacitation.

ISSCR standards for the use of human stem cells in basic research.

The laboratory culture of human stem cells seeks to capture a cellular state as an in vitro surrogate of a biological system. For the results and outputs from this research to be accurate, meaningful, and durable, standards that ensure reproducibility and reliability of the data should be applied. Although such standards have been previously proposed for repositories and distribution centers, no widely accepted best practices exist for laboratory research with human pluripotent and tissue stem cells. To fill that void, the International Society for Stem Cell Research has developed a set of recommendations, including reporting criteria, for scientists in basic research laboratories. These criteria are designed to be technically and financially feasible and, when implemented, enhance the reproducibility and rigor of stem cell research.

A contamination focused approach for optimizing the single-cell RNA-seq experiment.

Droplet-based single-cell RNA-seq (scRNA-seq) data are plagued by ambient contaminations caused by nucleic acid material released by dead and dying cells. This material is mixed into the buffer and is co-encapsulated with cells, leading to a lower signal-to-noise ratio. Although there exist computational methods to remove ambient contaminations post-hoc, the reliability of algorithms in generating high-quality data from low-quality sources remains uncertain. Here, we assess data quality before data filtering by a set of quantitative, contamination-based metrics that assess data quality more effectively than standard metrics. Through a series of controlled experiments, we report improvements that can minimize ambient contamination outside of tissue dissociation, via cell fixation, improved cell loading, microfluidic dilution, and nuclei versus cell preparation; many of these parameters are inaccessible on commercial platforms. We provide end-users with insights on factors that can guide their decision-making regarding optimizations that minimize ambient contamination, and metrics to assess data quality.

Robotic-assisted percutaneous pelvis fixation: A case report.

Key Clinical Message: It may be possible to extend the use of the robotic arm to pelvic and acetabular surgery leading to safe, repeatable screw placement, and less radiation exposure for patients, surgeons and OR staff. Abstract: In this case, a novel, robotic-assisted technique was used to place a sacroiliac screw in a patient with unstable injuries of the pelvic ring. Intraoperative and postoperative fluoroscopic, radiographic, and CT imaging demonstrated a safely positioned 6.5 mm cannulated screw without unplanned cortical violation or impingement on neurovascular structures. To our knowledge, this is the first such reported case using a robot widely available in the Americas or Europe.