Hypophosphatasia (HPP) is a rare, heritable metabolic disorder caused by deficient activity of tissue-nonspecific alkaline phosphatase (TNSALP). Asfotase alfa (AA) is a human recombinant TNSALP that ...promotes bone mineralization and is approved to treat eligible patients with HPP.
This prospective single-center observational study evaluated AA in adults with pediatric-onset HPP over 2 years of treatment (ClinicalTrials.govNCT03418389). Primary outcomes evaluated physical function; secondary outcomes assessed quality of life (QoL) and pain.
The study included 17 females and 5 males (mean age: 48.7 years). Median distance walked in the 6-Minute Walk Test increased significantly from baseline to 12 months (P = 0.034) and results were sustained. Median Timed Up and Go test time significantly decreased from baseline at 12 (P = 0.003) and 24 months (P = 0.005), as did the median chair rise time test at 12 (P = 0.003) and 24 months (P < 0.002). The change from baseline in usual gait speed was significant at 12 (P = 0.003) and 24 months (P = 0.015). Mean dominant and nondominant hand grip strength improved at 24 months (P = 0.029 and P = 0.019, respectively). Median Short Form 36 Physical Component Summary scores significantly improved from baseline at 12 (P = 0.012) and 24 (P = 0.005) months, and median Lower Extremity Functional Scale scores improved from baseline at 12 (P = 0.001) and 24 (P = 0.002) months. No significant change was noted in pain level at these timepoints. While injection site reactions occurred in 86.4 % of the participants, there were no severe side effects or safety findings.
Adults with pediatric-onset HPP treated with AA experienced marked improvement in functional and QoL outcomes that were observed as early as within 3 months of initial treatment and were sustained over 24 months.
•Asfotase alfa in adults with hypophosphatasia was evaluated over 24 months.•Patients had marked improvement in functional and quality of life outcomes.•Response of bone turnover markers suggests onset of bone tissue remodeling.
During the SARS-CoV-2 pandemic, dominant viral variants were repeatedly replaced by new variants with altered properties, frequently changing the dynamics of the infection event, as well as the ...effectiveness of vaccines and therapeutics. SARS-CoV-2 variant monitoring by whole genome sequencing was established at the University Medical Center Mainz, Germany to support patient management during the pandemic.
SARS-CoV-2 RNA samples from the University Medical Center were analysed weekly with whole genome sequencing. The genome sequences obtained were aligned with sequences from public databases to perform variant assignment. For classification purposes, phylogenetic trees were constructed to map the variant distribution in the clinical settings and the current outbreak events at that time. We describe the surveillance procedures using an example from a geriatric ward.
For monitoring, a time series was created covering two years of the pandemic. The changes from the Alpha to the Delta and the Omicron variants of SARS-CoV-2 could thus be precisely observed. The increasingly rapid switch of Omicron subvariants in the recent past could be tracked. The elucidation of phylogenetic relationships between circulating strains allowed conclusions about transmission pathways. Using an example from a geriatric ward, we demonstrated how variant monitoring by whole genome sequencing supported the infection prevention and control procedures on a ward and contribute to the control of outbreaks.
This example of SARS-CoV-2 demonstrates the effectiveness of targeted, local monitoring by molecular variant analysis. The program proved to be instrumental in controlling an outbreak on a geriatric ward.
In vorliegender Dissertation wurden die Langzeitergebnisse hinsichtlich der Lebensqualität der Patienten, die sich im Zeitraum 2002-2010 eine proximale Humerusfraktur zuzogen und mittels ...intramedullärer Drahtosteosynthese nach Kapandji im König Ludwig Haus in Würzburg operiert wurden, untersucht. Die Nachuntersuchung erfolgte mittels klinischer, radiologischer und sonografischer Untersuchung sowie durch Verwendung des DASH-Fragebogens, Constant Murley Score und der Visuellen Analogskala.
The outcome of patients which got a proximal humerus fracture betweeen 2002-2010 and had been treated with Kapandji pinning in König Ludwig Haus in Würzburg has been investigated in this dissertation. The follow up examination includes clinical investigation, X-ray and ultrasonics as well as DASH form, Constant Murley Score and the visual analogue scale.
The prime function of nucleoli is ribogenesis, however, several other, non-canonical functions have recently been identified, including a role in genotoxic stress response. Upon DNA damage, numerous ...proteins shuttle dynamically between the nucleolus and the nucleoplasm, yet the underlying molecular mechanisms are incompletely understood. Here, we demonstrate that PARP1 and PARylation contribute to genotoxic stress-induced nucleolar-nucleoplasmic shuttling of key genome maintenance factors in HeLa cells. Our work revealed that the RECQ helicase, WRN, translocates from nucleoli to the nucleoplasm upon treatment with the oxidizing agent H
O
, the alkylating agent 2-chloroethyl ethyl sulfide (CEES), and the topoisomerase inhibitor camptothecin (CPT). We show that after treatment with H
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and CEES, but not CPT, WRN translocation was dependent on PARP1 protein, yet independent of its enzymatic activity. In contrast, nucleolar-nucleoplasmic translocation of the base excision repair protein, XRCC1, was dependent on both PARP1 protein and its enzymatic activity. Furthermore, gossypol, which inhibits PARP1 activity by disruption of PARP1-protein interactions, abolishes nucleolar-nucleoplasmic shuttling of WRN, XRCC1 and PARP1, indicating the involvement of further upstream factors. In conclusion, this study highlights a prominent role of PARP1 in the DNA damage-induced nucleolar-nucleoplasmic shuttling of genome maintenance factors in HeLa cells in a toxicant and protein-specific manner.
Genetic and acquired abnormalities causing dysregulation of the complement alternative pathway contribute to atypical hemolytic uremic syndrome (aHUS), a rare disorder characterized by ...thrombocytopenia, nonimmune microangiopathic hemolytic anemia, and acute kidney failure. However, in a substantial proportion of patients the disease-associated alterations are still unknown.
Whole-exome and whole-genome sequencing were performed in two unrelated families with infantile recessive aHUS. Sequencing of cDNA from affected individuals was used to test for the presence of aberrant mRNA species. Expression of mutant diacylglycerol kinase epsilon (DGKE) protein was evaluated with western blotting.
Whole-exome sequencing analysis with conventional variant filtering parameters did not reveal any obvious candidate mutation in the first family. The report of aHUS-associated mutations in DGKE, encoding DGKE, led to re-examination of the noncoding DGKE variants obtained from next-generation sequencing, allowing identification of a novel intronic DGKE mutation (c.888+40A>G) that segregated with disease. Sequencing of cDNA from affected individuals revealed aberrant forms of DGKE mRNA predicted to cause profound abnormalities in the protein catalytic site. By whole-genome sequencing, the same mutation was found in compound heterozygosity with a second nonsense DGKE mutation in all affected siblings of another unrelated family. Homozygous and compound heterozygous patients presented similar clinical features, including aHUS presentation in the first year of life, multiple relapsing episodes, and proteinuria, which are prototypical of DGKE-associated aHUS.
This is the first report of a mutation located beyond the exon-intron boundaries in aHUS. Intronic mutations such as these are underreported because conventional filtering parameters used to process next-generation sequencing data routinely exclude these regions from downstream analyses in both research and clinical settings. The results suggest that analysis of noncoding regions of aHUS-associated genes coupled with mRNA sequencing might provide a tool to explain genetically unsolved aHUS cases.
Effective treatment of some types of cancer can be achieved by modulating cell lineage-specific rather than tumor-specific targets. We conducted a systematic search for novel agents selectively toxic ...to cells of hematopoietic origin. Chemical library screenings followed by hit-to-lead optimization identified OT-82, a small molecule with strong efficacy against hematopoietic malignancies including acute myeloblastic and lymphoblastic adult and pediatric leukemias, erythroleukemia, multiple myeloma, and Burkitt's lymphoma in vitro and in mouse xenograft models. OT-82 was also more toxic towards patients-derived leukemic cells versus healthy bone marrow-derived hematopoietic precursors. OT-82 was shown to induce cell death by inhibiting nicotinamide phosphoribosyltransferase (NAMPT), the rate-limiting enzyme in the salvage pathway of NAD synthesis. In mice, optimization of OT-82 dosing and dietary niacin further expanded the compound's therapeutic index. In toxicological studies conducted in mice and nonhuman primates, OT-82 showed no cardiac, neurological or retinal toxicities observed with other NAMPT inhibitors and had no effect on mouse aging or longevity. Hematopoietic and lymphoid organs were identified as the primary targets for dose limiting toxicity of OT-82 in both species. These results reveal strong dependence of neoplastic cells of hematopoietic origin on NAMPT and introduce OT-82 as a promising candidate for the treatment of hematological malignancies.
Background and purpose
Charcot–Marie–Tooth disease (CMT) is a heterogeneous group of disorders caused by mutations in at least 100 genes. However, approximately 60% of cases with axonal neuropathies ...(CMT2) still remain without a genetic diagnosis. We aimed at identifying novel disease genes responsible for CMT2.
Methods
We performed whole exome sequencing and targeted next generation sequencing panel analyses on a cohort of CMT2 families with evidence for autosomal recessive inheritance. We also performed functional studies to explore the pathogenetic role of selected variants.
Results
We identified rare, recessive variants in the MYO9B (myosin IX) gene in two families with CMT2. MYO9B has not yet been associated with a human disease. MYO9B is an unconventional single‐headed processive myosin motor protein with signaling properties, and, consistent with this, our results indicate that a variant occurring in the MYO9B motor domain impairs protein expression level and motor activity. Interestingly, a Myo9b‐null mouse has degenerating axons in sciatic nerves and optic nerves, indicating that MYO9B plays an essential role in both peripheral nervous system and central nervous system axons, respectively. The degeneration observed in the optic nerve prompted us to screen for MYO9B mutations in a cohort of patients with optic atrophy (OA). Consistent with this, we found compound heterozygous variants in one case with isolated OA.
Conclusions
Novel or very rare variants in MYO9B are associated with CMT2 and isolated OA.
► Laser treatment of bi-phasic Al/Al2O3 nanowires lead to dense and fully crystalline alumina (crack free). ► We present visible laser assisted ceramic synthesis instead of using CO2 laser. ► We ...present a dynamic surface which reflects the laser light after the phase transformation. ► Laser treated surface exhibits a hardness comparable to single crystalline alumina (sapphire).
Al/Al2O3 bi-phasic nanowires (Al-core/Al2O3 shell) are prepared by chemical vapor deposition (CVD) using single source precursor (SSP) approach. Such bi-phasic nanostructures were heat-treated using an argon laser operating at visible wavelengths. Al core seems to act as an active binder, which might decrease the inhomogeneous heating and thermal gradients. Nanoindentation method is used to estimate the hardness of the laser treated surfaces. Hardness values and pop-in behaviour in loading-curve indicate a formation of α-Al2O3 with very low defect density. It is believed that Al/Al2O3 bi-phasic layers exhibit a dynamic change by transforming into alumina after the laser irradiation and this leads to alteration of the optical absorption especially in the visible wavelength region. Following the full transformation to alumina, the surface reflects back the laser light which hinders inhomogeneous and excessive heating. In this context, laser treatment of Al/Al2O3 bi-phasic nanowires provides a controlled sintering process which can open up various applications in different fields.
The management of end stage heart failure patients is only possible by heart transplantation or by the implantation of artificial hearts as a bridge for later transplantation. However, these ...therapeutic strategies are limited by a lack of donor hearts and by the associated complications, such as coagulation and infection, due to the used artificial mechanical circulatory assist devices. Therefore, new strategies for myocardial regenerative approaches are under extensive research to produce contractile myocardial tissue in the future to replace non-contractile myocardial ischemic and scarred tissue. Different approaches, such as cell transplantation, have been studied intensively. Although successful approaches have been observed, there are still limitations to the application. It is envisaged that myocardial tissue engineering can be used to help replace infarcted non-contractile tissue. The developed tissue should later mimic the aligned fibrillar structure of the extracellular matrix and provide important guidance cues for the survival, function and the needed orientation of cardiomyocytes. Nanostructured surfaces have been tested to provide a guided direction that cells can follow. In the present study, the cellular adhesion/alignment of human cardiomyocytes and the biocompatibility have been investigated after cultivation on different laser-patterned nanowires compared with unmodified nanowires. As a result, the nanostructured surfaces possessed good biocompatibility before and after laser modification. The laser-induced scalability of the pattern enabled the growth and orientation of the adhered myocardial tissue. Such approaches may be used to modify the surface of potential scaffolds to develop myocardial contractile tissue in the future.
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