The case for treatment of monogenic SRNS with calcineurin inhibitors.

Currently, no evidence-based guidelines exist for treatment of children with monogenic steroid-resistant nephrotic syndrome. A retrospective study on 141 patients from Malakasioti et al. revealed that 27.6% responded to calcineurin inhibitor (CNI) treatment, and 75% of responders maintained stable kidney function. Virtually all CNI nonresponders developed progressive loss of kidney function. This study emphasized roles for CNIs in patients with monogenic steroid-resistant nephrotic syndrome, and the need for future studies to identify CNI response biomarkers.

Hiding in plain sight: genetics of childhood steroid-resistant nephrotic syndrome in Sub-Saharan Africa.

Steroid-resistant nephrotic syndrome (SRNS) is the most severe form of childhood nephrotic syndrome with an increased risk of progression to chronic kidney disease stage 5. Research endeavors to date have identified more than 80 genes that are associated with SRNS. Most of these genes regulate the structure and function of the podocyte, the visceral epithelial cells of the glomerulus. Although individuals of African ancestry have the highest prevalence of SRNS, especially those from Sub-Saharan Africa (SSA), with rates as high as 30-40% of all cases of nephrotic syndrome, studies focusing on the characterization and understanding of the genetic basis of SRNS in the region are negligible compared with Europe and North America. Therefore, it remains unclear if some of the variants in SRNS genes that are deemed pathogenic for SRNS are truly disease causing, and if the leading causes of monogenic nephrotic syndrome in other populations are the same for children in SSA with SRNS. Other implications of this lack of genetic data for SRNS in the region include the exclusion of children from the region from clinical trials aimed at identifying potential novel therapeutic agents for this severe form of nephrotic syndrome. This review underlines a need for concerted efforts to advance the genetic basis of SRNS in children in SSA. Such endeavors will complement global efforts at understanding the genetic basis of nephrotic syndrome.

Translating Outcomes from the Clinical Setting to Preclinical Models: Chronic Pain and Functionality in Chronic Musculoskeletal Pain.

Fibromyalgia (FM) is a chronic pain disorder characterized by chronic widespread musculoskeletal pain (CWP), resting pain, movement-evoked pain (MEP), and other somatic symptoms that interfere with daily functioning and quality of life. In clinical studies, this symptomology is assessed, while preclinical models of CWP are limited to nociceptive assays. The aim of the study was to investigate the human-to-model translatability of clinical behavioral assessments for spontaneous (or resting) pain and MEP in a preclinical model of CWP. For preclinical measures, the acidic saline model of FM was used to induce widespread muscle pain in adult female mice. Two intramuscular injections of acidic or neutral pH saline were administered following baseline measures, 5 days apart. An array of adapted evoked and spontaneous pain measures and functional assays were assessed for 3 weeks. A novel paradigm for MEP assessment showed increased spontaneous pain following activity. For clinical measures, resting and movement-evoked pain and function were assessed in adult women with FM. Moreover, we assessed correlations between the preclinical model of CWP and in women with fibromyalgia to examine whether similar relationships between pain assays that comprise resting and MEP existed in both settings. For both preclinical and clinical outcomes, MEP was significantly associated with mechanical pain sensitivity. Preclinically, it is imperative to expand how the field assesses spontaneous pain and MEP when studying multi-symptom disorders like FM. Targeted pain assessments to match those performed clinically is an important aspect of improving preclinical to clinical translatability of animal models.

A Rare Autosomal Dominant Variant in Regulator of Calcineurin Type 1 (RCAN1) Gene Confers Enhanced Calcineurin Activity and May Cause FSGS.

BACKGROUND: Podocyte dysfunction is the main pathologic mechanism driving the development of FSGS and other morphologic types of steroid-resistant nephrotic syndrome (SRNS). Despite significant progress, the genetic causes of most cases of SRNS have yet to be identified. METHODS: Whole-genome sequencing was performed on 320 individuals from 201 families with familial and sporadic NS/FSGS with no pathogenic mutations in any known NS/FSGS genes. RESULTS: Two variants in the gene encoding regulator of calcineurin type 1 (RCAN1) segregate with disease in two families with autosomal dominant FSGS/SRNS. In vitro, loss of RCAN1 reduced human podocyte viability due to increased calcineurin activity. Cells expressing mutant RCAN1 displayed increased calcineurin activity and NFAT activation that resulted in increased susceptibility to apoptosis compared with wild-type RCAN1. Treatment with GSK-3 inhibitors ameliorated this elevated calcineurin activity, suggesting the mutation alters the balance of RCAN1 regulation by GSK-3ß, resulting in dysregulated calcineurin activity and apoptosis. CONCLUSIONS: These data suggest mutations in RCAN1 can cause autosomal dominant FSGS. Despite the widespread use of calcineurin inhibitors in the treatment of NS, genetic mutations in a direct regulator of calcineurin have not been implicated in the etiology of NS/FSGS before this report. The findings highlight the therapeutic potential of targeting RCAN1 regulatory molecules, such as GSK-3ß, in the treatment of FSGS.

Laser spot size and scaling laws for laser beam additive manufacturing.

Laser powder bed fusion (L-PBF) additive manufacturing (AM) requires the careful selection of laser process parameters for each feedstock material and machine, which is a laborious process. Scaling laws based on the laser power, speed, and spot size; melt pool geometry; and thermophysical properties can potentially reduce this effort by transferring knowledge from one material and/or laser system to another. Laser spot size is one critical parameter that is less well studied for scaling laws compared to laser power and scan speed. Consequently, single track laser scans were generated with a spot size (D4σ) range of 50 µm to 322 µm and melt pool aspect ratio (depth over spot radius) range from 0.1 to 7.0. These were characterized by in-situ thermography, cross-sectioning, and optical microscopy. Scaling laws from literature were applied and evaluated based on melt pool depth predictions. Scaling laws that contain a minimum of three dimensionless parameters and account for changing absorption between conduction and keyhole mode provide the most accurate melt pool depth predictions (< 35 % difference from experiments), which is comparable to thermal simulation results from literature for a select number of cases.

Measurement Uncertainty of Surface Temperature Distributions for Laser Powder Bed Fusion Processes.

This paper describes advances in measuring the characteristic spatial distribution of surface temperature and emissivity during laser-metal interaction under conditions relevant for laser powder bed fusion (LPBF) additive manufacturing processes. Detailed descriptions of the measurement process, results, and approaches to determining uncertainties are provided. Measurement uncertainties have complex dependencies on multiple process parameters, so the methodology is demonstrated on one set of process parameters and one material. Well-established literature values for high-purity nickel solidification temperature and emissivity at the solidification temperature were used to evaluate the predicted uncertainty of the measurements. The standard temperature measurement uncertainty is found to be approximately 0.9% of the absolute temperature (16 AC), and the standard relative emissivity measurement uncertainty is found to be approximately 8% at the solidification point of high-purity nickel, both of which are satisfactory. This paper also outlines several potential sources of test uncertainties, which may require additional experimental evaluation. The largest of these are the metal vapor and ejecta that are produced as process by-products, which can potentially affect the imaging quality, reflectometry results, and thermal signature of the process, while also affecting the process of laser power delivery. Furthermore, the current paper focuses strictly on the uncertainties of the emissivity and temperature measurement approach and therefore does not detail a variety of uncertainties associated with experimental controls that must be evaluated for future generation of reference data.

Common risk variants in NPHS1 and TNFSF15 are associated with childhood steroid-sensitive nephrotic syndrome.

To understand the genetics of steroid-sensitive nephrotic syndrome (SSNS), we conducted a genome-wide association study in 987 childhood SSNS patients and 3,206 healthy controls with Japanese ancestry. Beyond known associations in the HLA-DR/DQ region, common variants in NPHS1-KIRREL2 (rs56117924, P=4.94E-20, odds ratio (OR) =1.90) and TNFSF15 (rs6478109, P=2.54E-8, OR=0.72) regions achieved genome-wide significance and were replicated in Korean, South Asian and African populations. Trans-ethnic meta-analyses including Japanese, Korean, South Asian, African, European, Hispanic and Maghrebian populations confirmed the significant associations of variants in NPHS1-KIRREL2 (Pmeta=6.71E-28, OR=1.88) and TNFSF15 (Pmeta=5.40E-11, OR=1.33) loci. Analysis of the NPHS1 risk alleles with glomerular NPHS1 mRNA expression from the same person revealed allele specific expression with significantly lower expression of the transcript derived from the risk haplotype (Wilcox test p=9.3E-4). Because rare pathogenic variants in NPHS1 cause congenital nephrotic syndrome of the Finnish type (CNSF), the present study provides further evidence that variation along the allele frequency spectrum in the same gene can cause or contribute to both a rare monogenic disease (CNSF) and a more complex, polygenic disease (SSNS).

The Human FSGS-Causing ANLN R431C Mutation Induces Dysregulated PI3K/AKT/mTOR/Rac1 Signaling in Podocytes.

BACKGROUND: We previously reported that mutations in the anillin (ANLN) gene cause familial forms of FSGS. ANLN is an F-actin binding protein that modulates podocyte cell motility and interacts with the phosphoinositide 3-kinase (PI3K) pathway through the slit diaphragm adaptor protein CD2-associated protein (CD2AP). However, it is unclear how the ANLN mutations cause the FSGS phenotype. We hypothesized that the R431C mutation exerts its pathogenic effects by uncoupling ANLN from CD2AP. METHODS: We conducted in vivo complementation assays in zebrafish to determine the effect of the previously identified missense ANLN variants, ANLNR431C and ANLNG618C during development. We also performed in vitro functional assays using human podocyte cell lines stably expressing wild-type ANLN (ANLNWT ) or ANLNR431C . RESULTS: Experiments in anln-deficient zebrafish embryos showed a loss-of-function effect for each ANLN variant. In human podocyte lines, expression of ANLNR431C increased cell migration, proliferation, and apoptosis. Biochemical characterization of ANLNR431C -expressing podocytes revealed hyperactivation of the PI3K/AKT/mTOR/p70S6K/Rac1 signaling axis and activation of mTOR-driven endoplasmic reticulum stress in ANLNR431C -expressing podocytes. Inhibition of mTOR, GSK-3ß, Rac1, or calcineurin ameliorated the effects of ANLNR431C . Additionally, inhibition of the calcineurin/NFAT pathway reduced the expression of endogenous ANLN and mTOR. CONCLUSIONS: The ANLNR431C mutation causes multiple derangements in podocyte function through hyperactivation of PI3K/AKT/mTOR/p70S6K/Rac1 signaling. Our findings suggest that the benefits of calcineurin inhibition in FSGS may be due, in part, to the suppression of ANLN and mTOR. Moreover, these studies illustrate that rational therapeutic targets for familial FSGS can be identified through biochemical characterization of dysregulated podocyte phenotypes.

On thermal properties of metallic powder in laser powder bed fusion additive manufacturing.

Powder thermal properties play a critical role in laser powder-bed fusion (LPBF) additive manufacturing, specifically, the reduced effective thermal conductivity compared to that of the solid significantly affects heat conduction, which can influence the melt pool characteristics, and consequently, the part mechanical properties. This study intends to indirectly measure the thermal conductivity of metallic powder, nickel-based super alloy 625 (IN625) and Ti-6Al-4V (Ti64), in LPBF using a combined approach that consists of laser flash analysis, finite element (FE) heat transfer modeling and a multivariate inverse method. The test specimens were designed and fabricated by a LPBF system to encapsulate powder in a hollow disk to imitate powder-bed conditions. The as-built specimens were then subjected to laser flash testing to measure the transient thermal response. Next, an FE model replicate the hollow disk samples and laser flash testing was developed. A multi-point optimization algorithm was used to inversely extract the thermal conductivity of LPBF powder from the FE model based on the measured transient thermal response. The results indicate that the thermal conductivity of IN625 powder used in LPBF ranges from 0.65 W/(m·K) to 1.02 W/(m·K) at 100 °C and 500 °C, respectively, showing a linear relationship with the temperature. On the other hand, Ti64 powder has a lower thermal conductivity than IN625 powder, about 35% to 40% smaller. However, the thermal conductivity ratio of the powder to the respective solid counterpart is quite similar between the two materials, about 4.2% to 6.9% for IN625 and 3.4% to 5.2% for Ti64.

A Combined Experimental-Numerical Method to Evaluate Powder Thermal Properties in Laser Powder Bed Fusion.

Powder bed metal additive manufacturing (AM) utilizes a high-energy heat source scanning at the surface of a powder layer in a predefined area to be melted and solidified to fabricate parts layer by layer. It is known that powder bed metal AM is primarily a thermal process, and further, heat conduction is the dominant heat transfer mode in the process. Hence, understanding the powder bed thermal conductivity is crucial to process temperature predictions, because powder thermal conductivity could be substantially different from its solid counterpart. On the other hand, measuring the powder thermal conductivity is a challenging task. The objective of this study is to investigate the powder thermal conductivity using a method that combines a thermal diffusivity measurement technique and a numerical heat transfer model. In the experimental aspect, disk-shaped samples, with powder inside, made by a laser powder bed fusion (LPBF) system, are measured using a laser flash system to obtain the thermal diffusivity and the normalized temperature history during testing. In parallel, a finite element (FE) model is developed to simulate the transient heat transfer of the laser flash process. The numerical model was first validated using reference material testing. Then, the model is extended to incorporate powder enclosed in an LPBF sample with thermal properties to be determined using an inverse method to approximate the simulation results to the thermal data from the experiments. In order to include the powder particles' contribution in the measurement, an improved model geometry, which improves the contact condition between powder particles and the sample solid shell, has been tested. A multipoint optimization inverse heat transfer method is used to calculate the powder thermal conductivity. From this study, the thermal conductivity of a nickel alloy 625 powder in powder bed conditions is estimated to be 1.01 W/m K at 500°C. [DOI: 10.1115/1.4040877].

Application of Finite Element, Phase-field, and CALPHAD-based Methods to Additive Manufacturing of Ni-based Superalloys.

Numerical simulations are used in this work to investigate aspects of microstructure and microseg-regation during rapid solidification of a Ni-based superalloy in a laser powder bed fusion additive manufacturing process. Thermal modeling by finite element analysis simulates the laser melt pool, with surface temperatures in agreement with in situ thermographic measurements on Inconel 625. Geometric and thermal features of the simulated melt pools are extracted and used in subsequent mesoscale simulations. Solidification in the melt pool is simulated on two length scales. For the multicomponent alloy Inconel 625, microsegregation between dendrite arms is calculated using the Scheil-Gulliver solidification model and DICTRA software. Phase-field simulations, using Ni-Nb as a binary analogue to Inconel 625, produced microstructures with primary cellular/dendritic arm spacings in agreement with measured spacings in experimentally observed microstructures and a lesser extent of microsegregation than predicted by DICTRA simulations. The composition profiles are used to compare thermodynamic driving forces for nucleation against experimentally observed precipitates identified by electron and X-ray diffraction analyses. Our analysis lists the precipitates that may form from FCC phase of enriched interdendritic compositions and compares these against experimentally observed phases from 1 h heat treatments at two temperatures: stress relief at 1143 K (870 °C) or homogenization at 1423 K (1150 °C).

Identifying uncertainty in laser powder bed fusion additive manufacturing models.

As additive manufacturing (AM) matures, models are beginning to take a more prominent stage in design and process planning for AM. A limitation frequently encountered in AM models is a lack of indication about their precision and accuracy. Often overlooked, information on model uncertainty is required for validation of AM models, qualification of AM-produced parts, and uncertainty management. This paper presents a discussion on the origin and propagation of uncertainty in laser powder bed fusion (L-PBF) models. Four sources of uncertainty are identified: modeling assumptions, unknown simulation parameters, numerical approximations, and measurement error in calibration data. Techniques to quantify uncertainty in each source are presented briefly, along with estimation algorithms to diminish prediction uncertainty with the incorporation of online measurements. The methods are illustrated with a case study based on a transient, stochastic thermal model designed for melt pool width predictions. Model uncertainty is quantified for single track experiments and the effect of online estimation in overhanging structures is studied via simulation. The application of these concepts to estimation and control of the L-PBF process is suggested.

Thermographic Measurements of the Commercial Laser Powder Bed Fusion Process at NIST.

Measurement of the high-temperature melt pool region in the laser powder bed fusion (L-PBF) process is a primary focus of researchers to further understand the dynamic physics of the heating, melting, adhesion, and cooling which define this commercially popular additive manufacturing process. This paper will detail the design, execution, and results of high speed, high magnification in-situ thermographic measurements conducted at the National Institute of Standards and Technology (NIST) focusing on the melt pool region of a commercial L-PBF process. Multiple phenomena are observed including plasma plume and hot particle ejection from the melt region. The thermographic measurement process will be detailed with emphasis on the 'measurability' of observed phenomena and the sources of measurement uncertainty. Further discussion will relate these thermographic results to other efforts at NIST towards L-PBF process finite element simulation and development of in-situ sensing and control methodologies.

A prospective study of microscope-integrated intraoperative fluorescein videoangiography during arteriovenous malformation surgery: preliminary results.

OBJECT: The authors report on the use of a recently developed microscope-integrated fluorescent module using low-dose intravenous fluorescein for videoangiography during arteriovenous malformation (AVM) surgery. METHODS: The authors analyzed the application of a low-dose intraoperative fluorescein in 4 consecutive patients undergoing AVM surgery. The ability to distinguish the associated vessels of the AVM from normal vessels and to assess the degree of AVM obliteration based on videoangiography of venous drainage was specifically analyzed. RESULTS: All 4 patients underwent fluorescein angiography without complication. In each case, videoangiography confirmed recognition of feeding arteries and draining veins through the operating oculars under the fluorescent mode. In one case involving a large frontal AVM, videoangiography demonstrated mainly cortical veins on the surface of the AVM and alerted the senior author to first tackle the feeding arteries in the interhemispheric space. While evaluating the flow within the different draining veins after most of the AVM was disconnected, videoangiography also prioritized the order for disconnection of large draining veins to allow mobilization the AVM and exposure of the remaining deep arterial feeders. In the other 3 cases, videoangiography allowed easy recognition of the angioarchitecture of the AVMs, estimated its cortical boundaries, and most importantly, assessed the flow within the draining veins before their disconnection. CONCLUSIONS: The authors found fluorescein videoangiography to be a useful adjunct in resection of AVMs. This technology offers the unique ability to visualize fluorescent vessels and nonfluorescent tissues in near-natural colors simultaneously and permits microsurgical manipulation of relevant structures under the fluorescent mode. Larger-scale studies are needed to establish its efficacy and wider applicability.

Cross-Sectional Melt Pool Geometry of Laser Scanned Tracks and Pads on Nickel Alloy 718 for the 2022 Additive Manufacturing Benchmark Challenges.

The Additive Manufacturing Benchmark Series (AM Bench) is a NIST-led organization that provides a continuing series of additive manufacturing benchmark measurements, challenge problems, and conferences with the primary goal of enabling modelers to test their simulations against rigorous, highly controlled additive manufacturing benchmark measurement data. To this end, single-track (1D) and pad (2D) scans on bare plate nickel alloy 718 were completed with thermography, cross-sectional grain orientation and local chemical composition maps, and cross-sectional melt pool size measurements. The laser power, scan speed, and laser spot size were varied for single tracks, and the scan direction was varied for pads. This article focuses on the cross-sectional melt pool size measurements and presents the predictions from challenge problems. Single-track depth correlated with volumetric energy density while width did not (within the studied parameters). The melt pool size for pad scans was greater than single tracks due to heat buildup. Pad scan melt pool depth was reduced when the laser scan direction and gas flow direction were parallel. The melt pool size in pad scans showed little to no trend against position within the pads. Uncertainty budgets for cross-sectional melt pool size from optical micrographs are provided for the purpose of model validation.

Genomic analysis of human brain metastases treated with stereotactic radiosurgery reveals unique signature based on treatment failure.

Stereotactic radiosurgery (SRS) has been shown to be efficacious for the treatment of limited brain metastasis (BM); however, the effects of SRS on human brain metastases have yet to be studied. We performed genomic analysis on resected brain metastases from patients whose resected lesion was previously treated with SRS. Our analyses demonstrated for the first time that patients possess a distinct genomic signature based on type of treatment failure including local failure, leptomeningeal spread, and radio-necrosis. Examination of the center and peripheral edge of the tumors treated with SRS indicated differential DNA damage distribution and an enrichment for tumor suppressor mutations and DNA damage repair pathways along the peripheral edge. Furthermore, the two clinical modalities used to deliver SRS, LINAC and GK, demonstrated differential effects on the tumor landscape even between controlled primary sites. Our study provides, in human, biological evidence of differential effects of SRS across BM's.