Raman spectroscopic study of anhydrous and hydrous REE phosphates, oxides, and hydroxides.

Rare earth elements (REE) include the lanthanides (La-Lu), Y, and Sc which are critical elements for the green energy transition. The REE show a decrease in ionic radii with increased atomic numbers, which results in a so-called lanthanide contraction systematically affecting crystal structures and mineral properties. Here we present a compilation of reference Raman spectra of ten REE sesquioxides (A-, B- and C-type), five REE hydroxides, eight xenotime-structured REE phosphate endmembers and two solid solutions, seven monazite-structured REE phosphate endmembers and two solid solutions and seven rhabdophane endmembers with up to five Ce1-xLREEx rhabdophane solid solutions (LREE = La-Gd). Raman mode assignment is based on a detailed literature review summarizing existing analytical work and theoretical calculations and systematic trends observed in this study by analyzing different REE-bearing solids. The wavenumbers of the main REE-O Raman band systematically increase with decreasing ionic radii forming discrete linear trends within isostructural mineral groups, that can be used to estimate the REE-O mode in other solids with known REE-O coordination numbers. Photoluminescence using 266 nm, 532 nm and 633 nm excitation laser wavelengths for REE-bearing oxides, hydroxides, anhydrous and hydrous phosphates is also presented providing a new framework for identifying REE-phases in phosphate-bearing natural mineral deposits.

Calreticulin and JAK2V617F driver mutations induce distinct mitotic defects in myeloproliferative neoplasms.

Myeloproliferative neoplasms (MPNs) encompass a diverse group of hematologic disorders driven by mutations in JAK2, CALR, or MPL. The prevailing working model explaining how these driver mutations induce different disease phenotypes is based on the decisive influence of the cellular microenvironment and the acquisition of additional mutations. Here, we report increased levels of chromatin segregation errors in hematopoietic cells stably expressing CALRdel52 or JAK2V617F mutations. Our investigations employing murine 32DMPL and human erythroleukemic TF-1MPL cells demonstrate a link between CALRdel52 or JAK2V617F expression and a compromised spindle assembly checkpoint (SAC), a phenomenon contributing to error-prone mitosis. This defective SAC is associated with imbalances in the recruitment of SAC factors to mitotic kinetochores upon CALRdel52 or JAK2V617F expression. We show that JAK2 mutant CD34 + MPN patient-derived cells exhibit reduced expression of the master mitotic regulators PLK1, aurora kinase B, and PP2A catalytic subunit. Furthermore, the expression profile of mitotic regulators in CD34 + patient-derived cells allows to faithfully distinguish patients from healthy controls, as well as to differentiate primary and secondary myelofibrosis from essential thrombocythemia and polycythemia vera. Altogether, our data suggest alterations in mitotic regulation as a potential driver in the pathogenesis in MPN.

The Effective Use of Anti-obesity Medications.

Obesity is a heterogeneous disease and there is wide patient-to-patient variability in response to all anti-obesity treatments including lifestyle modifications, anti-obesity medications (AOMs), devices, and bariatric surgery. To effectively treat obesity, practitioners must be knowledgeable about all of these treatment modalities including on-label and off-label AOMs. Care should be individualized to the patient taking into consideration their unique challenges with weight loss, their goals, the presence of comorbidities, medication contraindications, and drug-drug interactions. There is currently no way to know which AOM will be most effective for a patient without trial and error; therefore, prescribe AOMs in sequence and consider combination therapy for optimal results. This article reviews the efficacy, safety, prescribing information, and other considerations for all of the currently available AOMs.

Altered Serum Acylcarnitines Profile after a Prolonged Stay in Intensive Care.

A stay in intensive care unit (ICU) exposes patients to a risk of carnitine deficiency. Moreover, acylated derivates of carnitine (acylcarnitines, AC) are biomarkers for metabolic mitochondrial dysfunction that have been linked to post-ICU disorders. This study aimed to describe the AC profile of survivors of a prolonged ICU stay (≥7 days). Survivors enrolled in our post-ICU clinic between September 2020 and July 2021 were included. Blood analysis was routinely performed during the days after ICU discharge, focusing on metabolic markers and including AC profile. Serum AC concentrations were determined by LC-MS/MS and were compared to the reference ranges (RR) established from serum samples of 50 non-hospitalized Belgian adults aged from 18 to 81 years. A total 162 patients (65.4% males, age 67 (58.7−73) years) survived an ICU stay of 9.7 (7.1−19.3) days and were evaluated 5 (3−8) days after discharge. Their AC profile was significantly different compared to RR, mostly in terms of short chain AC: the sum of C3, C4 and C5 derivates reached 1.36 (0.98−1.99) and 0.86 (0.66−0.99) µmol/L respectively (p < 0.001). Free carnitine (C0) concentration of survivors (46.06 (35.04−56.35) µmol/L) was similar to RR (43.64 (36.43−52.96) µmol/L) (p = 0.55). C0 below percentile 2.5 of RR was observed in 6/162 (3.7%) survivors. Their total AC/C0 ratio was 0.33 (0.22−0.42). A ratio above 0.4 was observed in 45/162 (27.8%) patients. In ICU survivors, carnitine deficiency was rare, but AC profile was altered and AC/C0 ratio was abnormal in more than 25%. The value of AC profile as a marker of post-ICU dysmetabolism needs further investigations.

An In Vitro Evaluation of the Biological and Osteogenic Properties of Magnesium-Doped Bioactive Glasses for Application in Bone Tissue Engineering.

Magnesium (Mg2+) is known to play a crucial role in mineral and matrix metabolism of bone tissue and is thus increasingly considered in the field of bone tissue engineering. Bioactive glasses (BGs) offer the promising possibility of the incorporation and local delivery of therapeutically active ions as Mg2+. In this study, two Mg2+-doped derivatives of the ICIE16-BG composition (49.46 SiO2, 36.27 CaO, 6.6 Na2O, 1.07 P2O5, 6.6 K2O (mol%)), namely 6Mg-BG (49.46 SiO2, 30.27 CaO, 6.6 Na2O, 1.07 P2O5, 6.6 K2O, 6.0 MgO (mol%) and 3Mg-BG (49.46 SiO2, 33.27 CaO, 6.6 Na2O, 1.07 P2O5, 6.6 K2O, 3.0 MgO (mol%)) were examined. Their influence on viability, proliferation and osteogenic differentiation of human mesenchymal stromal cells (MSCs) was explored in comparison to the original ICIE16-BG. All BGs showed good biocompatibility. The Mg2+-doped BGs had a positive influence on MSC viability alongside with inhibiting effects on MSC proliferation. A strong induction of osteogenic differentiation markers was observed, with the Mg2+-doped BGs significantly outperforming the ICIE16-BG regarding the expression of genes encoding for protein members of the osseous extracellular matrix (ECM) at certain observation time points. However, an overall Mg2+-induced enhancement of the expression of genes encoding for ECM proteins could not be observed, possibly due to a too moderate Mg2+ release. By adaption of the Mg2+ release from BGs, an even stronger impact on the expression of genes encoding for ECM proteins might be achieved. Furthermore, other BG-types such as mesoporous BGs might provide a higher local presence of the therapeutically active ions and should therefore be considered for upcoming studies.

Outbreak or pseudo-outbreak? Integrating SARS-CoV-2 sequencing to validate infection control practices in a dialysis facility.

BACKGROUND: The COVID-19 pandemic poses a particularly high risk for End Stage Renal Disease (ESRD) patients so rapid identification of case clusters in ESRD facilities is essential. Nevertheless, with high community prevalence, a series of ESRD patients may test positive contemporaneously for reasons unrelated to their shared ESRD facility. Here we describe a series of 5 cases detected within 11 days in November 2020 in a hospital-based 32-station ESRD facility in Southwest Wisconsin, the subsequent facility-wide testing, and the use of genetic sequence analysis to evaluate links between cases. METHODS: Four patient cases and one staff case were identified in symptomatic individuals by RT-PCR. Facility-wide screening was conducted using rapid SARS-CoV-2 antigen tests. SARS-CoV-2 genome sequences were obtained from residual diagnostic specimens. RESULTS: Facility-wide screening of 47 staff and 107 patients identified no additional cases. Residual specimens from 4 of 5 cases were available for genetic sequencing. Clear genetic differences proved that these contemporaneous cases were not linked. CONCLUSIONS: With high community prevalence, epidemiological data alone is insufficient to deem a case cluster an outbreak. Cluster evaluation with genomic data, when available with a short turn-around time, can play an important role in infection prevention and control response programs.

A rare case of diffuse alveolar hemorrhage caused by acute mycoplasma pneumoniae pneumonia.

Mycoplasma pneumoniae is a common bacterial pathogen that causes atypical community-acquired pneumonia. Illness onset can be gradual and progressive over weeks. Patients typically have cough, pharyngitis, malaise, and tracheobronchitis. Although symptoms are frequently mild, the initial presentation can be severe with numerous complications. We present a case of a 28-year-old male who presented with 1 day of significant hemoptysis. He was intubated for airway protection and underwent bronchoscopy, which showed multiple blood clots in several lung lobes, consistent with diffuse alveolar hemorrhage (DAH). His workup was negative for pulmonary embolism, coagulopathy, and vasculitis. He tested positive for rhinovirus and mycoplasma pneumoniae IgM (negative IgG). He was ultimately discharged home with oral doxycycline to complete a 10-day course. DAH is a rare presentation and life-threatening complication of mycoplasma pneumonia. Although there is a reported association between DAH and rhinovirus, our patient improved with antibiotics making mycoplasma pneumoniae the likely culprit. When encountering hemoptysis or alveolar bleeding, clinicians should have low suspicion for atypical infections and start appropriate antibiotics early in the clinical course.

Development of Neuroleptic Malignant Syndrome in a Patient with Lewy Body Dementia after Intramuscular Administration of Paliperidone.

Neuroleptic malignant syndrome (NMS) is a potentially fatal diagnosis composed of hyperpyrexia, muscle rigidity, altered mental status, and autonomic instability. This syndrome has significant systemic complications including acute renal failure, rhabdomyolysis, hyperkalemia, and seizure. It is associated with the use of both typical and atypical antipsychotics. Due to the extensive neurodegenerative destruction of dopaminergic and acetylcholinergic pathways, patients with Lewy body dementia (LBD) are particularly sensitive to antidopaminergic and anticholinergic medications, making them more susceptible to extrapyramidal side effects and NMS. We present a case of a 72-year-old female with LBD who developed muscular rigidity, vital sign instability, and altered mental status after receiving one dose of paliperidone palmitate injection two weeks prior to admission. Initial blood work was unrevealing. Extensive workup including EEG, lumbar puncture with cerebrospinal fluid analysis, and brain MRI was unremarkable. She was treated with seven days of bromocriptine and a lorazepam taper with improvement in muscle rigidity. However, her mental status never improved, and she remained comatose. She was later intubated for airway protection after an aspiration event that led to hypoxia. Her respiratory status never recovered, and she was transitioned to comfort care. This case demonstrates the complexity and potential fatality of NMS. Clinicians should be aware of this dangerous complication of antipsychotic use in patients with LBD as these patients may be more susceptible to this complication.

Quantum Chemical Calculation of Molecular and Periodic Peptide and Protein Structures.

Special-purpose classical force fields (FFs) provide good accuracy at very low computational cost, but their application is limited to systems for which potential energy functions are available. This excludes most metal-containing proteins or those containing cofactors. In contrast, the GFN2-xTB semiempirical quantum chemical method is parametrized for almost the entire periodic table. The accuracy of GFN2-xTB is assessed for protein structures with respect to experimental X-ray data. Furthermore, the results are compared with those of two special-purpose FFs, HF-3c, PM6-D3H4X, and PM7. The test sets include proteins without any prosthetic groups as well as metalloproteins. Crystal packing effects are examined for a set of smaller proteins to validate the molecular approach. For the proteins without prosthetic groups, the special purpose FF OPLS-2005 yields the smallest overall RMSD to the X-ray data but GFN2-xTB provides similarly good structures with even better bond-length distributions. For the metalloproteins with up to 5000 atoms, a good overall structural agreement is obtained with GFN2-xTB. The full geometry optimizations of protein structures with on average 1000 atoms in wall-times below 1 day establishes the GFN2-xTB method as a versatile tool for the computational treatment of various biomolecules with a good accuracy/computational cost ratio.

Osteogenic properties of manganese-doped mesoporous bioactive glass nanoparticles.

Mesoporous bioactive glass nanoparticles (MBGNs) based on the SiO2 -P2 O5 -CaO system have demonstrated promising properties for the local delivery of therapeutically active ions with the aim to improve their osteogenic properties. Manganese (Mn) has been identified as a candidate ion for local application in bone tissue engineering applications. It remains unknown how SiO2 -P2 O5 -CaO-based MBGNs influence human bone marrow-derived mesenchymal stromal cells (BMSCs) in terms of viability, proliferation, and differentiation and how these features can be modified by the addition of Mn to the MBGNs' composition. Therefore, in this study, MBGNs (composition in mol%: 50 SiO2 , 40 CaO, 10 P2 O5 ) and its Mn-doped derivate 5Mn-MBGNs (composition in mol%: 50 SiO2 , 35 CaO, 10 P2 O5 , 5 MnO) were applied to a culture of BMSCs in two different concentrations. With increasing concentration, 5Mn-MBGNs supported osteogenic differentiation and enhanced the upregulation of genes encoding for extracellular matrix proteins but also negatively influenced cell viability and proliferation. When applied in lower concentrations, MBGNs showed not only viability- and growth-enhancing effects but also significant pro-osteogenic features-however, these positive properties deteriorated with increasing concentration. Two major conclusions can be drawn from this study: (a) supplementation with Mn enhances the osteogenic properties of MBGNs in a dose-dependent manner and (b) MBGNs constitute an attractive vector for therapeutically active ions since it exhibits an intrinsic pro-osteogenic potential that can be improved and/or modified by incorporation of therapeutically active ions. Future studies should focus on the evaluation of further candidate ions that are known to influence osteogenic differentiation positively.

Bioactive Glass (BG) ICIE16 Shows Promising Osteogenic Properties Compared to Crystallized 45S5-BG.

The ICIE16-bioactive glass (BG) (48.0 SiO2, 6.6 Na2O, 32.9 CaO, 2.5 P2O5, 10.0 K2O (wt %)) has been developed as an alternative to 45S5-BG, the original BG composition (45.0 SiO2, 24.5 Na2O, 24.5 CaO, 6.0 P2O5 (wt %)), with the intention of broadening the BG sintering window while maintaining bioactivity. Because there is a lack of reports on ICIE16-BG biological properties, the influence of ICIE16-BG on viability, proliferation, and osteogenic differentiation of human mesenchymal stromal cells (MSCs) was evaluated in direct comparison to 45S5-BG in this study. The BGs underwent heat treatment similar to that which is required in order to fabricate scaffolds by sintering, which resulted in crystallization of 45S5-BG (45S5-CBG) while ICIE16 remained amorphous. Granules based on both BGs were biocompatible, but ICIE16-BG was less harmful to cell viability, most likely due to a more pronounced pH alkalization in the 45S5-CBG group. ICIE16-BG outperformed 45S5-CBG in terms of osteogenic differentiation at the cellular level, as determined by the increased activity of alkaline phosphatase. However, granules from both BGs were comparable regarding the stimulation of expression levels of genes encoding for osseous extracellular matrix (ECM) proteins. The addition of therapeutically active ions to ICIE16-BG might further improve its ability to stimulate ECM production and should be investigated in upcoming studies.

Characterization of Single Amino Acid Variations in an EDTA-Tolerating Non-specific Nuclease from the Ice-Nucleating Bacterium Pseudomonas syringae.

Non-specific nuclease (NSN) can be applied in industrial downstream processing to remove nucleic acids from crude protein extracts or in cell-sorting systems to degrade nucleic acids derived from lysed cells. PsNuc from the ice-nucleating bacterium Pseudomonas syringae has the ability to decompose double- and single-stranded DNA in linear or circular form and RNA. It is not affected by the presence of metal-ion chelators such as EDTA and tolerates several protease inhibitors and reducing agents. A multiple sequence alignment of PsNuc with closely related enzymes (97-99% identity on the protein level) within the family Pseudomonaceae revealed the presence of only six amino acid residues that are variable in putative NSN from different members of the genus Pseudomonas. Single amino acid variants were produced in recombinant form in Escherichia coli, purified, and characterized. They showed similar activity compared to PsNuc, but a single variant even displayed an improved performance with an activity of > 20,000 U/mg at 35 °C, while amino acid residues S148 and V161 were found to be essential for enzymatic functionality. These results suggest that homologous nucleases from Pseudomonaceae display high activity levels in a metal-ion-independent manner and are therefore of interest for applications in biotechnology.

Comparative analysis of two non-specific nucleases of the phospholipase D family from the plant pathogen competitor bacterium Pantoea agglomerans.

Bacterial non-specific nucleases of the phospholipase D family are widely distributed among the members of the Enterobacteriaceae. Each genome mainly contains a single copy of a gene encoding a phospholipase D family protein. However, two distantly related isozymes (< 40% identity at the protein level) were identified by BLAST-analyses in the plant pathogenic competitor enterobacterium Pantoea agglomerans. The two nucleases PaNuc-1 and PaNuc-2 were produced in Escherichia coli. Identical gene constructs and expression conditions resulted in the production of PaNuc-1 in soluble form, while PaNuc-2 remained insoluble in inclusion bodies. PaNuc-2 was refolded and both proteins were purified by a combination of affinity and ion exchange chromatography. Proteolytic removal of the HIS-tag allowed the characterization of pure and mature tag-less proteins. Enzymatic properties of both isozymes revealed that they are non-specific nucleases, displaying activities against RNA, single- and double-stranded genomic DNA as well as circular plasmids. However, their biochemical activity profiles were clearly different, with PaNuc-1 being optimally active at 70 °C and pH 7.0, while PaNuc-2 was most active at 45 °C and pH 7.0. The enzymes retained > 90% nuclease activity at EDTA concentrations of 4 mM (PaNuc-2) and 20 mM (PaNuc-1), respectively. Different enzymatic properties suggest that the roles of PaNuc-1 and PaNuc-2 differ in the cell and might be the result of functional diversification after an ancient gene duplication event took place. The fact that both enzymes could be easily produced in recombinant form and their tolerance against metal ion chelators in combination with a broad substrate promiscuity might pave the way to versatile biotechnological applications.

A non-specific nucleolytic enzyme and its application potential in EDTA-containing buffer solutions.

OBJECTIVES: Metal-ion independent non-specific nucleases are of high potential for applications in EDTA-containing bioprocessing workflows. RESULTS: A novel extracellular non-specific nuclease EcNuc from the enterobacterium Escherichia coli has been identified. The recombinant gene was expressed and the protein was purified. Maximum activity of the enzyme was detected at 41.7 °C and at an acidic pH of 5.8. EcNuc tolerates EDTA in the reaction buffer at concentrations of up to 20 mM and the activity is not impaired by high concentrations of mono- and divalent metal ions in the absence of EDTA. The viscosity of crude protein extracts after cell lysis in EDTA-containing buffers is reduced when supplemented with EcNuc. CONCLUSION: Proof-of-concept has been demonstrated that a metal-ion independent non-specific nuclease can be applied for removal of nucleic acids in EDTA-containing buffers for the subsequent purification of proteins from crude extracts.

Seven-year itch: a perplexing case of lichen planus-lupus erythematosus overlap syndrome.

Lichen planus-lupus erythematosus overlap syndrome is a rare disorder characterized by clinical and histopathological features of both lichen planus (LP) and lupus erythematosus (LE). Cutaneous lesions commonly affect the distal arms, legs, face, and trunk and these plaques are often large, scaly, painful, and atrophic, often exhibiting hypopigmentation or a red to blue-violet color. We report a case of LP-LE overlap syndrome diagnosed in a man previously believed to have atypical lichen planus who presented with an exacerbation of exuberant pruritic erythematous scaly plaques. The patient had six separate skin biopsies all of which displayed features of LP. Because the clinical symptoms did not correlate to the histopathological picture, a seventh skin biopsy with direct immunofluorescence (DIF) was performed and immunologic markers measured. The DIF demonstrated early lupus bands; serologic testing exhibited elevated ANA and anti-SSA. These findings established the diagnosis of LP-LE overlap syndrome. The patient was started on hydroxychloroquine with short-term trials of oral prednisone during disease flares, which took place in the first three months of treatment.

A tandem CD19/CD20 CAR lentiviral vector drives on-target and off-target antigen modulation in leukemia cell lines.

BACKGROUND: Clinical success with chimeric antigen receptor (CAR)- based immunotherapy for leukemia has been accompanied by the associated finding that antigen-escape variants of the disease are responsible for relapse. To target hematologic malignancies with a chimeric antigen receptor (CAR) that targets two antigens with a single vector, and thus potentially lessen the chance of leukemic escape mutations, a tandem-CAR approach was investigated. METHODS: Antigen binding domains from the FMC63 (anti-CD19) and Leu16 (anti-CD20) antibodies were linked in differing configurations to transmembrane and T cell signaling domains to create tandem-CARs. Expression on the surface of primary human T cells was induced by transduction with a single lentiviral vector (LV) encoding the tandem-CAR. Tandem-CARs were compared to single antigen targeting CARs in vitro and in vivo, and to an admixture of transduced cells expressing each CAR in vivo in immunodeficient (NSG) disease-bearing mice. RESULTS: Tandem constructs efficient killed the Raji leukemia cell line both in vitro and in vivo. Tandem CARs generated less cytokine than the CD20 CAR, but similar to CD19 CARs, on their own. In co-culture experiments at low effector to target ratios with both single- and tandem- CAR-T cells, a rapid down-modulation of full-length CD19 expression was seen on leukemia targets. There also was a partial down-modulation of CD22, and to a lesser degree, of CD20. Our data also highlight the extreme sensitivity of the NALM-6 cell line to general lymphocyte-mediated cytotoxicity. While single and tandem constructs were effective in vivo in a standard setting, in a high-disease burden setting, the tandem CAR proved both effective and less toxic than an admixture of transduced T cell populations expressing single CARs. CONCLUSION: Tandem CARs are equally effective in standard disease models to single antigen specificity CARs, and may be both more effective and less toxic in a higher disease burden setting. This may be due to optimized cell killing with more moderate cytokine production. The rapid co-modulation of CD19, CD20, and CD22 may account for the ability to rapidly evolve escape mutants by selecting for leukemic clones that not require these target antigens for continued expansion.

Assessment of the extent of starch dissolution in dimethyl sulfoxide by 1H NMR spectroscopy.

Complete dissolution is needed for the separation, characterization, or homogeneous labeling of whole starch molecules. A method is presented to quantify the extent of starch dissolution in DMSO for the first time; it is validated on a commercial rice starch. It is used directly on starch dispersions containing possible undissolved or co-dissolved species. High-amylose maize starches, known to be digested slowly in vivo, only quantitatively dissolve in the presence of high concentrations of an H-bond disrupter, LiBr, although they form clear dispersions at low LiBr concentrations. Starch quantitatively dissolves from waxy rice flours; non-starch components partially co-dissolve but do not interfere with the dissolution quantification.