Plasmodium falciparum Cysteine-Rich Protective Antigen (CyRPA) Elicits Detectable Levels of Invasion-Inhibitory Antibodies during Natural Infection in Humans.

Plasmodium falciparum cysteine-rich protective antigen (CyRPA) is a conserved component of an essential erythrocyte invasion complex (RH5/Ripr/CyRPA) and a target of potent cross-strain parasite-neutralizing antibodies. While naturally acquired human RH5 antibodies have been functionally characterized, there are no similar reports on CyRPA. Thus, we analyzed the parasite-neutralizing activity of naturally acquired human CyRPA antibodies. In this regard, CyRPA human antibodies were measured and purified from malaria-infected plasma obtained from patients in central India and analyzed for their parasite neutralizing activity via in vitro growth inhibition assays (GIA). We report that, despite being susceptible to antibodies, CyRPA is a highly conserved antigen that does not appear to be under substantial immune selection pressure, as a very low acquisition rate for anti-CyRPA antibodies was reported in malaria-exposed Indians. We demonstrate for the first time that the small amounts of natural CyRPA antibodies exhibited functional parasite-neutralizing activity and that a CyRPA-based vaccine formulation induces highly potent antibodies in rabbits. Importantly, the vaccine-induced CyRPA antibodies exhibited a robust 50% inhibitory concentration (IC50) of 21.96 µg/ml, which is comparable to the IC50 of antibodies against the leading blood-stage vaccine candidate, reticulocyte-binding-like homologous protein 5 (RH5). Our data support CyRPA as a unique vaccine target that is highly susceptible to immune attack but is highly conserved compared to other leading candidates such as MSP-1 and AMA-1, further substantiating its promise as a leading blood-stage vaccine candidate.

Treg sensitivity to FasL and relative IL-2 deprivation drive idiopathic aplastic anemia immune dysfunction.

Idiopathic aplastic anemia (AA) has 2 key characteristics: an autoimmune response against hematopoietic stem/progenitor cells and regulatory T-cells (Tregs) deficiency. We have previously demonstrated reduction in a specific subpopulation of Treg in AA, which predicts response to immunosuppression. The aims of the present study were to define mechanisms of Treg subpopulation imbalance and identify potential for therapeutic intervention. We have identified 2 mechanisms that lead to skewed Treg composition in AA: first, FasL-mediated apoptosis on ligand interaction; and, second, relative interleukin-2 (IL-2) deprivation. We have shown that IL-2 augmentation can overcome these mechanisms. Interestingly, when high concentrations of IL-2 were used for in vitro Treg expansion cultures, AA Tregs were able to expand. The expanded populations expressed a high level of p-BCL-2, which makes them resistant to apoptosis. Using a xenograft mouse model, the function and stability of expanded AA Tregs were tested. We have shown that these Tregs were able to suppress the macroscopic clinical features and tissue manifestations of T-cell-mediated graft-versus-host disease. These Tregs maintained their suppressive properties as well as their phenotype in a highly inflammatory environment. Our findings provide an insight into the mechanisms of Treg reduction in AA. We have identified novel targets with potential for therapeutic interventions. Supplementation of ex vivo expansion cultures of Tregs with high concentrations of IL-2 or delivery of IL-2 directly to patients could improve clinical outcomes in addition to standard immunosuppressive therapy.

Antibody Combinations Targeting the Essential Antigens CyRPA, RH5, and MSP-119 Potently Neutralize Plasmodium falciparum Clinical Isolates From India and Africa.

BACKGROUND: Targeting multiple key antigens that mediate distinct Plasmodium falciparum erythrocyte invasion pathways is an attractive approach for the development of blood-stage malaria vaccines. However, the challenge is to identify antigen cocktails that elicit potent strain-transcending parasite-neutralizing antibodies efficacious at low immunoglobulin G concentrations feasible to achieve through vaccination. Previous reports have screened inhibitory antibodies primarily against well adapted laboratory parasite clones. However, validation of the parasite-neutralizing efficacy against clinical isolates with minimal in vitro cultivation is equally significant to better ascertain their prospective in vivo potency. METHODS: We evaluated the parasite-neutralizing activity of different antibodies individually and in combinations against laboratory adapted clones and clinical isolates. Clinical isolates were collected from Central India and Mozambique, Africa, and characterized for their invasion properties and genetic diversity of invasion ligands. RESULTS: In our portfolio, we evaluated 25 triple antibody combinations and identified the MSP-Fu+CyRPA+RH5 antibody combination to elicit maximal parasite neutralization against P. falciparum clinical isolates with variable properties that underwent minimal in vitro cultivation. CONCLUSIONS: The MSP-Fu+CyRPA+RH5 combination exhibited highly robust parasite neutralization against P. falciparum clones and clinical isolates, thus substantiating them as promising candidate antigens and establishing a proof of principle for the development of a combinatorial P. falciparum blood-stage malaria vaccine.

Deep phenotyping of Tregs identifies an immune signature for idiopathic aplastic anemia and predicts response to treatment.

Idiopathic aplastic anemia (AA) is an immune-mediated and serious form of bone marrow failure. Akin to other autoimmune diseases, we have previously shown that in AA regulatory T cells (Tregs) are reduced in number and function. The aim of this study was to further characterize Treg subpopulations in AA and investigate the potential correlation between specific Treg subsets and response to immunosuppressive therapy (IST) as well as their in vitro expandability for potential clinical use. Using mass cytometry and an unbiased multidimensional analytical approach, we identified 2 specific human Treg subpopulations (Treg A and Treg B) with distinct phenotypes, gene expression, expandability, and function. Treg B predominates in IST responder patients, has a memory/activated phenotype (with higher expression of CD95, CCR4, and CD45RO within FOXP3(hi), CD127(lo) Tregs), expresses the interleukin-2 (IL-2)/STAT5 pathway and cell-cycle commitment genes. Furthermore, in vitro-expanded Tregs become functional and take on the characteristics of Treg B. Collectively, this study identifies human Treg subpopulations that can be used as predictive biomarkers for response to IST in AA and potentially other autoimmune diseases. We also show that Tregs from AA patients are IL-2-sensitive and expandable in vitro, suggesting novel therapeutic approaches such as low-dose IL-2 therapy and/or expanded autologous Tregs and meriting further exploration.

Somatic mutations identify a subgroup of aplastic anemia patients who progress to myelodysplastic syndrome.

The distinction between acquired aplastic anemia (AA) and hypocellular myelodysplastic syndrome (hMDS) is often difficult, especially nonsevere AA. We postulated that somatic mutations are present in a subset of AA, and predict malignant transformation. From our database, we identified 150 AA patients with no morphological evidence of MDS, who had stored bone marrow (BM) and constitutional DNA. We excluded Fanconi anemia, mutations of telomere maintenance, and a family history of BM failure (BMF) or cancer. The initial cohort of 57 patients was screened for 835 known genes associated with BMF and myeloid cancer; a second cohort of 93 patients was screened for mutations in ASXL1, DNMT3A, BCOR, TET2, and MPL. Somatic mutations were detected in 19% of AA, and included ASXL1 (n = 12), DNMT3A (n = 8) and BCOR (n = 6). Patients with somatic mutations had a longer disease duration (37 vs 8 months, P < .04), and shorter telomere lengths (median length, 0.9 vs 1.1, P < .001), compared with patients without mutations. Somatic mutations in AA patients with a disease duration of >6 months were associated with a 40% risk of transformation to MDS (P < .0002). Nearly one-fifth of AA patients harbor mutations in genes typically seen in myeloid malignancies that predicted for later transformation to MDS.

Utility of peripheral blood for cytogenetic and mutation analysis in myelodysplastic syndrome.

Recent studies have shown that more than 80% of bone marrow (BM) samples from patients with myelodysplastic syndrome (MDS) harbor somatic mutations and/or genomic aberrations, which are of diagnostic and prognostic importance. We investigated the potential use of peripheral blood (PB) and serum to identify and monitor BM-derived genetic markers using high-resolution single nucleotide polymorphism array (SNP-A) karyotyping and parallel sequencing of 22 genes frequently mutated in MDS. This pilot study showed a 100% SNP-A karyotype concordance and a 97% mutation concordance between the BM and PB. In contrast, mutation analysis using Sanger sequencing of PB and serum-derived DNA showed only 65% and 42% concordance to BM, respectively. Our results show the potential utility of PB as a surrogate for BM for MDS patients, thus avoiding the need for repeated BM aspirates particularly in elderly patients and those with fibrotic or hypocellular marrows.

Functional characterization of CD4+ T cells in aplastic anemia.

The role of CD4(+) T cells in the pathogenesis of aplastic anemia (AA) is not well characterized. We investigate CD4(+) T-cell subsets in AA. Sixty-three patients with acquired AA were studied. Th1 and Th2 cells were significantly higher in AA patients than in healthy donors (HDs; P = .03 and P = .006). Tregs were significantly lower in patients with severe AA than in HDs (P < .001) and patients with non-severe AA (P = .01). Th17 cells were increased in severe AA (P = .02) but normal in non-severe AA. Activated and resting Tregs were reduced in AA (P = .004; P = .01), whereas cytokine-secreting non-Tregs were increased (P = .003). Tregs from AA patients were unable to suppress normal effector T cells. In contrast, AA effector T cells were suppressible by Tregs from HDs. Th1 clonality in AA, investigated by high-throughput sequencing, was greater than in HDs (P = .03). Our results confirm that Th1 and Th2 cells are expanded and Tregs are functionally abnormal in AA. The clonally restricted expansion of Th1 cells is most likely to be antigen-driven, and induces an inflammatory environment, that exacerbate the functional impairment of Tregs, which are reduced in number.

Predicting Mechanical Properties of Polymer Materials Using Rate-Dependent Material Models: Finite Element Analysis of Bespoke Upper Limb Orthoses.

Three-dimensional printing-especially with fused deposition modeling (FDM)-is widely used in the medical field as it enables customization. FDM is versatile owing to the availability of various materials, but selecting the appropriate material for a certain application can be challenging. Understanding materials' mechanical behaviors, particularly those of polymeric materials, is vital to determining their suitability for a given application. Physical testing with universal testing machines is the most used method for determining the mechanical behaviors of polymers. This method is resource-intensive and requires cylinders for compression testing and unique dumbbell-shaped specimens for tensile testing. Thus, a specialized fixture must be designed to conduct mechanical testing for the customized orthosis, which is costly and time-consuming. Finite element (FE) analysis using an appropriate material model must be performed to identify the mechanical behaviors of a customized shape (e.g., an orthosis). This study analyzed three material models, namely the Bergström-Boyce (BB), three-network (TN), and three-network viscoplastic (TNV) models, to determine the mechanical behaviors of polymer materials for personalized upper limb orthoses and examined three polymer materials: PLA, ABS, and PETG. The models were first calibrated for each material using experimental data. Once the models were calibrated and found to fit the data appropriately, they were employed to examine the customized orthosis's mechanical behaviors through FE analysis. This approach is innovative in that it predicts the mechanical characteristics of a personalized orthosis by combining theoretical and experimental investigations.

An Insight into the Characteristics of 3D Printed Polymer Materials for Orthoses Applications: Experimental Study.

Knee orthoses assist patients with impaired gait through the amendment of knee abnormalities, restoration of mobility, alleviation of pain, shielding, and immobilization. The inevitable issues with laborious traditional plaster molding procedures for orthoses can be resolved with 3D printing. However, a number of challenges have limited the adoption of 3D printing, the most significant of which is the proper material selection for orthoses. This is so because the material used to make an orthosis affects its strength, adaptability, longevity, weight, moisture response, etc. This study intends to examine the mechanical, physical, and dimensional characteristics of three-dimensional (3D) printing materials (PLA, ABS, PETG, TPU, and PP). The aim of this investigation is to gain knowledge about these materials' potential for usage as knee orthosis materials. Tensile testing, Olympus microscope imaging, water absorption studies, and coordinate measuring machine-based dimension analysis are used to characterize the various 3D printing materials. Based on the investigation, PLA outperforms all other materials in terms of yield strength (25.98 MPa), tensile strength (30.89 MPa), and shrinkage (0.46%). PP is the least water absorbent (0.15%) and most flexible (407.99%); however, it is the most difficult to fabricate using 3D printing. When producing knee orthoses with 3D printing, PLA can be used for the orthosis frame and other structural elements, PLA or ABS for moving parts like hinges, PP for padding, and TPU or PP for the straps. This study provides useful information for scientists and medical professionals who are intrigued about various polymer materials for 3D printing and their effective utilization to fabricate knee orthoses.

Assessment of consolidative multi-criteria decision making (C-MCDM) algorithms for optimal mapping of polymer materials in additive manufacturing: A case study of orthotic application.

Objective: The objectives of this research are twofold. The primary goal is to introduce, investigate, and contrast consolidative multi-criteria decision-making (C-MCDM) approaches. The second objective is the investigation of five alternative additive manufacturing materials. Methods: It integrates the subjective and objective weights using the Bayes hypothesis in conjunction with a normal method. Chang's Extent Analysis Method under fuzzy logic is used to estimate subjective weights and the CRITIC approach is used for assessing objective weights. Ranking techniques, including the simple ranking process (SRP), multi-objective optimization based on ratio analysis (MOORA), measurement alternatives and ranking according to compromise solution (MARCOS), and technique for order preference by similarity to ideal solution (TOPSIS) are applied. It also encompasses sensitivity analysis based on Kendall's coefficient of concordance and rank reversal phenomenon analysis. Spearman's rank correlation coefficient, a weighted rank measure of correlation, and rank similarity coefficient are among the metrics used to evaluate agreement between different approaches. It entails gathering expert opinions regarding the importance of various criteria as well as conducting extensive experiments. Results: The findings of the study indicate that polylactic acid is the best material to use for orthoses. When compared to the other MCDM approaches being discussed, SRP is the most reliable approach. It is also demonstrated that the SRP, MARCOS, and TOPSIS methods are rank reversal-free. Furthermore, SRP exhibits a very poor association with the TOPSIS technique but a strong correlation with the MOORA and MARCOS approaches. Conclusions: To ensure results reliability, it is necessary to consider both the subjectivity and objectivity of weights as well as apply multiple MCDM methodologies in addition to sensitivity analysis.

Influence of donor-recipient sex on engraftment of normal and leukemia stem cells in xenotransplantation.

Immunodeficient mouse models are widely used for the assessment of human normal and leukemic stem cells. Despite the advancements over the years, reproducibility, as well as the differences in the engraftment of human cells in recipient mice remains to be fully resolved. Here, we used various immunodeficient mouse models to characterize the effect of donor-recipient sex on the engraftment of the human leukemic and healthy cells. Donor human cells and recipient immunodeficient mice demonstrate sex-specific engraftment levels with significant differences observed in the lineage output of normal CD34+ hematopoietic stem and progenitor cells upon xenotransplantation. Intriguingly, human female donor cells display heightened sensitivity to the recipient mice's gender, influencing their proliferation and resulting in significantly increased engraftment in female recipient mice. Our study underscores the intricate interplay taking place between donor and recipient characteristics, shedding light on important considerations for future studies, particularly in the context of pre-clinical research.

TP53 mutations in myelodysplastic syndrome are strongly correlated with aberrations of chromosome 5, and correlate with adverse prognosis.

This study aimed to determine the incidence/prognostic impact of TP53 mutation in 318 myelodysplastic syndrome (MDS) patients, and to correlate the changes to cytogenetics, single nucleotide polymorphism array karyotyping and clinical outcome. The median age was 65 years (17-89 years) and median follow-up was 45 months [95% confidence interval (CI) 27-62 months]. TP53 mutations occurred in 30 (9.4%) patients, exclusively in isolated del5q (19%) and complex karyotype (CK) with -5/5q-(72%), correlated with International Prognostic Scoring System intermediate-2/high, TP53 protein expression, higher blast count and leukaemic progression. Patients with mutant TP53 had a paucity of mutations in other genes implicated in myeloid malignancies. Median overall survival of patients with TP53 mutation was shorter than wild-type (9 versus 66 months, P < 0.001) and it retained significance in multivariable model (Hazard Ratio 3.8, 95%CI 2.3-6.3,P < 0.001). None of the sequentially analysed samples showed a disappearance of the mutant clone or emergence of new clones, suggesting an early occurrence of TP53 mutations. A reduction in mutant clone correlated with response to 5-azacitidine, however clones increased in non-responders and persisted at relapse. The adverse impact of TP53 persists after adjustment for cytogenetic risk and is of practical importance in evaluating prognosis. The relatively common occurrence of these mutations in two different prognostic spectrums of MDS, i.e. isolated 5q- and CK with -5/5q-, possibly implies two different mechanistic roles for TP53 protein.

Spliceosome mutations exhibit specific associations with epigenetic modifiers and proto-oncogenes mutated in myelodysplastic syndrome.

The recent identification of acquired mutations in key components of the spliceosome machinery strongly implicates abnormalities of mRNA splicing in the pathogenesis of myelodysplastic syndromes. However, questions remain as to how these aberrations functionally combine with the growing list of mutations in genes involved in epigenetic modification and cell signaling/transcription regulation identified in these diseases. In this study, amplicon sequencing was used to perform a mutation screen in 154 myelodysplastic syndrome patients using a 22-gene panel, including commonly mutated spliceosome components (SF3B1, SRSF2, U2AF1, ZRSR2), and a further 18 genes known to be mutated in myeloid cancers. Sequencing of the 22-gene panel revealed that 76% (n=117) of the patients had mutations in at least one of the genes, with 38% (n=59) having splicing gene mutations and 49% (n=75) patients harboring more than one gene mutation. Interestingly, single and specific epigenetic modifier mutations tended to coexist with SF3B1 and SRSF2 mutations (P<0.03). Furthermore, mutations in SF3B1 and SRSF2 were mutually exclusive to TP53 mutations both at diagnosis and at the time of disease transformation. Moreover, mutations in FLT3, NRAS, RUNX1, CCBL and C-KIT were more likely to co-occur with splicing factor mutations generally (P<0.02), and SRSF2 mutants in particular (P<0.003) and were significantly associated with disease transformation (P<0.02). SF3B1 and TP53 mutations had varying impacts on overall survival with hazard ratios of 0.2 (P<0.03, 95% CI, 0.1-0.8) and 2.1 (P<0.04, 95% CI, 1.1-4.4), respectively. Moreover, patients with splicing factor mutations alone had a better overall survival than those with epigenetic modifier mutations, or cell signaling/transcription regulator mutations with and without coexisting mutations of splicing factor genes, with worsening prognosis (P<0.001). These findings suggest that splicing factor mutations are maintained throughout disease evolution with emerging oncogenic mutations adversely affecting patients' outcome, implicating spliceosome mutations as founder mutations in myelodysplastic syndromes.

Finite Element Analysis of Upper Limb Splint Designs and Materials for 3D Printing.

Three-dimensional (3D) printed splints must be lightweight and adequately ventilated to maximize the patient's convenience while maintaining requisite strength. The ensuing loss of strength has a substantial impact on the transformation of a solid splint model into a perforated or porous model. Thus, two methods for making perforations-standard approach and topological optimization-are investigated in this study. The objective of this research is to ascertain the impact of different perforation shapes and their distribution as well as topology optimization on the customized splint model. The solid splint models made of various materials have been transformed into porous designs to evaluate their strength by utilizing Finite Element (FE) simulation. This study will have a substantial effect on the designing concept for medical devices as well as other industries such as automobiles and aerospace. The novelty of the research refers to creating the perforations as well as applying topology optimization and 3D printing in practice. According to the comparison of the various materials, PLA had the least amount of deformation and the highest safety factor for all loading directions. Additionally, it was shown that all perforation shapes behave similarly, implying that the perforation shape's effect is not notably pronounced. However, square perforations seemed to perform the best out of all the perforation shape types. It was also obvious that the topology-optimized hand splint outperformed that with square perforations. The topology-optimized hand splint weighs 26% less than the solid splint, whereas the square-perforated hand splint weighs roughly 12% less. Nevertheless, the user must choose which strategy (standard perforations or topology optimization) to employ based on the available tools and prerequisites.

Design, Analysis, and 3D Printing of a Patient-Specific Polyetheretherketone Implant for the Reconstruction of Zygomatic Deformities.

The reconstruction of craniomaxillofacial deformities, especially zygomatic bone repair, can be exigent due to the complex anatomical structure and the sensitivity of the crucial organs involved. The need to reconstruct the zygomatic bone in the most precise way is of crucial importance for enhancing the patient outcomes and health care-related quality of life (HRQL). Autogenous bone grafts, despite being the gold standard, do not match bone forms, have limited donor sites and bone volume, and can induce substantial surgical site morbidity, which may lead to adverse outcomes. The goal of this study is to provide an integrated approach that includes various processes, from patient scanning to implant manufacture, for the restoration of zygomatic bone abnormalities utilizing Polyetheretherketone (PEEK) material, while retaining adequate aesthetic and facial symmetry. This study takes an integrated approach, including computer-aided implant design using the mirror reconstruction technique, investigating the biomechanical behavior of the implant under loading conditions, and carrying out a fitting accuracy analysis of the PEEK implant fabricated using state-of-the-art additive manufacturing technology. The findings of the biomechanical analysis results reveal the largest stress of approximately 0.89 MPa, which is relatively low in contrast to the material's yield strength and tensile strength. A high degree of sturdiness in the implant design is provided by the maximum value of strain and deformation, which is also relatively low at roughly 2.2 × 10-4 and 14 µm. This emphasizes the implant's capability for load resistance and safety under heavy loading. The 3D-printed PEEK implant observed a maximum deviation of 0.4810 mm in the outside direction, suggesting that the aesthetic result or the fitting precision is adequate. The 3D-printed PEEK implant has the potential to supplant the zygoma bone in cases of severe zygomatic reconstructive deformities, while improving the fit, stability, and strength of the implant.

Polyether-Ether-Ketone (PEEK) and Its 3D-Printed Quantitate Assessment in Cranial Reconstruction.

Three-dimensional (3D) printing, medical imaging, and implant design have all advanced significantly in recent years, and these developments may change how modern craniomaxillofacial surgeons use patient data to create tailored treatments. Polyether-ether-ketone (PEEK) is often seen as an attractive option over metal biomaterials in medical uses, but a solid PEEK implant often leads to poor osseointegration and clinical failure. Therefore, the objective of this study is to demonstrate the quantitative assessment of a custom porous PEEK implant for cranial reconstruction and to evaluate its fitting accuracy. The research proposes an efficient process for designing, fabricating, simulating, and inspecting a customized porous PEEK implant. In this study, a CT scan is utilized in conjunction with a mirrored reconstruction technique to produce a skull implant. In order to foster cell proliferation, the implant is modified into a porous structure. The implant's strength and stability are examined using finite element analysis. Fused filament fabrication (FFF) is utilized to fabricate the porous PEEK implants, and 3D scanning is used to test its fitting accuracy. The results of the biomechanical analysis indicate that the highest stress observed was approximately 61.92 MPa, which is comparatively low when compared with the yield strength and tensile strength of the material. The implant fitting analysis demonstrates that the implant's variance from the normal skull is less than 0.4436 mm, which is rather low given the delicate anatomy of the area. The results of the study demonstrate the implant's endurance while also increasing the patient's cosmetic value.

Modeling Consequences of COVID-19 and Assessing Its Epidemiological Parameters: A System Dynamics Approach.

In 2020, coronavirus (COVID-19) was declared a global pandemic and it remains prevalent today. A necessity to model the transmission of the virus has emerged as a result of COVID-19's exceedingly contagious characteristics and its rapid propagation throughout the world. Assessing the incidence of infection could enable policymakers to identify measures to halt the pandemic and gauge the required capacity of healthcare centers. Therefore, modeling the susceptibility, exposure, infection, and recovery in relation to the COVID-19 pandemic is crucial for the adoption of interventions by regulatory authorities. Fundamental factors, such as the infection rate, mortality rate, and recovery rate, must be considered in order to accurately represent the behavior of the pandemic using mathematical models. The difficulty in creating a mathematical model is in identifying the real model variables. Parameters might vary significantly across models, which can result in variations in the simulation results because projections primarily rely on a particular dataset. The purpose of this work was to establish a susceptible-exposed-infected-recovered (SEIR) model describing the propagation of the COVID-19 outbreak throughout the Kingdom of Saudi Arabia (KSA). The goal of this study was to derive the essential COVID-19 epidemiological factors from actual data. System dynamics modeling and design of experiment approaches were used to determine the most appropriate combination of epidemiological parameters and the influence of COVID-19. This study investigates how epidemiological variables such as seasonal amplitude, social awareness impact, and waning time can be adapted to correctly estimate COVID-19 scenarios such as the number of infected persons on a daily basis in KSA. This model can also be utilized to ascertain how stress (or hospital capacity) affects the percentage of hospitalizations and the number of deaths. Additionally, the results of this study can be used to establish policies or strategies for monitoring or restricting COVID-19 in Saudi Arabia.

Performance Evaluation of Input Power of Diode Laser on Machined Leather Specimen in Laser Beam Cutting Process.

Numerous industries, including footwear, handicrafts, and the automobile industry, utilize leather materials. The main goal of this study was to investigate the effect of input power of the diode laser in laser cutting on vegetable chrome tanned buffalo leather to enhance the cutting process. In the present investigation, carbonization, kerf width, and material removal rate (MRR) were taken as performance measures. The diode-based laser beam machining was designed and fabricated with 2.5 W, 5.5 W, and 20 W diode laser to cut vegetable chrome tanned leather. The high-intensity 20 W diode laser produced lower carbonization, lower kerf width, and higher material removal rate compared with the 2.5 W and 5.5 W diodes. This improved performance was due to the adjustable features associated with this diode laser actuation in the form of circular shape with adjustable diameter. A high power with a lower spot size under pulsed mode can produce higher power density. Since a higher power density can establish less interaction time, it produces lower carbonization. Due to the ability of the 20 W diode laser driver to control the beam shape and size, it could produce a lower kerf width and higher MRR. The optimal parameters for cutting chrome vegetable tanned cow leather were a standoff distance of 18 mm, feed rate of 200 mm/min, and duty cycle of 70%.

Vitamin B5 and succinyl-CoA improve ineffective erythropoiesis in SF3B1-mutated myelodysplasia.

Patients with myelodysplastic syndrome and ring sideroblasts (MDS-RS) present with symptomatic anemia due to ineffective erythropoiesis that impedes their quality of life and increases morbidity. More than 80% of patients with MDS-RS harbor splicing factor 3B subunit 1 (SF3B1) mutations, the founder aberration driving MDS-RS disease. Here, we report how mis-splicing of coenzyme A synthase (COASY), induced by mutations in SF3B1, affects heme biosynthesis and erythropoiesis. Our data revealed that COASY was up-regulated during normal erythroid differentiation, and its silencing prevented the formation of erythroid colonies, impeded erythroid differentiation, and precluded heme accumulation. In patients with MDS-RS, loss of protein due to COASY mis-splicing led to depletion of both CoA and succinyl-CoA. Supplementation with COASY substrate (vitamin B5) rescued CoA and succinyl-CoA concentrations in SF3B1mut cells and mended erythropoiesis differentiation defects in MDS-RS primary patient cells. Our findings reveal a key role of the COASY pathway in erythroid maturation and identify upstream and downstream metabolites of COASY as a potential treatment for anemia in patients with MDS-RS.

Next-generation sequencing of the TET2 gene in 355 MDS and CMML patients reveals low-abundance mutant clones with early origins, but indicates no definite prognostic value.

Mutations in the TET2 gene are frequent in myeloid disease, although their biologic and prognostic significance remains unclear. We analyzed 355 patients with myelodysplastic syndromes using "next-generation" sequencing for TET2 aberrations, 91 of whom were also subjected to single-nucleotide polymorphism 6.0 array karyotyping. Seventy-one TET2 mutations, with a relative mutation abundance (RMA) ≥ 10%, were identified in 39 of 320 (12%) myelodysplastic syndrome and 16 of 35 (46%) chronic myelomonocytic leukemia patients (P < .001). Interestingly, 4 patients had multiple mutations likely to exist as independent clones or on alternate alleles, suggestive of clonal evolution. "Deeper" sequencing of 96 patient samples identified 4 additional mutations (RMA, 3%-6.3%). Importantly, TET2 mutant clones were also found in T cells, in addition to CD34(+) and total bone marrow cells (23.5%, 38.5%, and 43% RMA, respectively). Only 20% of the TET2-mutated patients showed loss of heterozygosity at the TET2 locus. There was no difference in the frequency of genome-wide aberrations, TET2 expression, or the JAK2V617F 46/1 haplotype between TET2-mutated and nonmutated patients. There was no significant prognostic association between TET2 mutations and World Health Organization subtypes, International Prognostic Scoring System score, cytogenetic status, or transformation to acute myeloid leukemia. On multivariate analysis, age (> 50 years) was associated with a higher incidence of TET2 mutation (P = .02).