| 1. |
- Attebäck, Maria, et al.
(författare)
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Formulation Optimization of Extemporaneous Oral Liquids Containing Naloxone and Propranolol for Pediatric Use
- 2022
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Ingår i: Scientia pharmaceutica. - : MDPI. - 0036-8709 .- 2218-0532. ; 90:1
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Tidskriftsartikel (refereegranskat)abstract
- There is a need to develop dosage forms suitable for children to improve drug treatment. Extemporaneous compounding of drugs for children is one way to meet these needs. However, excipients generally considered as safe in adults may not be appropriate in dosage forms intended for children. The aim was to optimize the composition of two pediatric liquid preparations by substituting paraben as a microbiological preservative and ethanol as a solubilizer, with excipients more suitable for pediatric use. The oral liquids were Naloxone 1 mg/mL and Propranolol 10 mg/mL. Twelve different formulations were tested with propranolol and naloxone, respectively, during the screening process to select appropriate formulations. Sodium benzoate and glycerol were used as a preservative and solubilizer, respectively, and different pH of the formulations were evaluated. The formulations were characterized according to dispensed dose (dosing accuracy), viscosity and osmolality. The optimized formulations from the screening process were tested with two amounts of sodium benzoate and microbiological assays were performed. These formulations were shown to have satisfactory preservative properties and dosing accuracy. The results showed that the oral liquids could be prepared without the addition of solubilizer and with lower osmolality (naloxone), thus reducing the risk of gastrointestinal side effects.
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| 2. |
- Bèchet, Nicholas B., et al.
(författare)
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Light sheet fluorescence microscopy of optically cleared brains for studying the glymphatic system
- 2020
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Ingår i: Journal of Cerebral Blood Flow and Metabolism. - 0271-678X. ; 40:10, s. 1975-1986
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Tidskriftsartikel (refereegranskat)abstract
- Fluid transport in the perivascular space by the glia-lymphatic (glymphatic) system is important for the removal of solutes from the brain parenchyma, including peptides such as amyloid-beta which are implicated in the pathogenesis of Alzheimer’s disease. The glymphatic system is highly active in the sleep state and under the influence of certain of anaesthetics, while it is suppressed in the awake state and by other anaesthetics. Here we investigated whether light sheet fluorescence microscopy of whole optically cleared murine brains was capable of detecting glymphatic differences in sleep- and awake-mimicking anaesthesia, respectively. Using light-sheet imaging of whole brains, we found anaesthetic-dependent cerebrospinal fluid (CSF) influx differences, including reduced tracer influx along tertiary branches of the middle cerebral artery and reduced influx along dorsal and anterior penetrating arterioles, in the awake-mimicking anaesthesia. This study establishes that light sheet microscopy of optically cleared brains is feasible for quantitative analyses and can provide images of the entire glymphatic system in whole brains.
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| 3. |
- Birtele, Marcella, et al.
(författare)
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Single-cell transcriptional and functional analysis of dopaminergic neurons in organoid-like cultures derived from human fetal midbrain
- 2022
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Ingår i: Development: For advances in developmental biology and stem cells. - : The Company of Biologists. - 1477-9129. ; 149:23
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Tidskriftsartikel (refereegranskat)abstract
- Significant efforts are ongoing to develop refined differentiation protocols to generate midbrain dopamine (DA) neurons from pluripotent stem cells (PSCs) for application in disease modeling, diagnostics, drug screening, and cell-based therapies for Parkinson's Disease (PD). An increased understanding of the timing and molecular mechanisms promoting the generation of distinct subtypes of human midbrain DA during development will be essential for guiding future efforts to generate molecularly defined and subtype-specific DA neurons from PSCs. Here, we used droplet-based single-cell RNA sequencing to transcriptionally profile the developing human ventral midbrain (VM) when the DA neurons are generated (6-11 weeks post-conception) and their subsequent differentiation into functional mature DA neurons in primary fetal 3D organoid-like cultures. This approach revealed that 3D cultures are superior to monolayer conditions for their ability to generate and maintain mature DA neurons; hence they have the potential to be used for studying human VM development. These results provide a unique transcriptional profile of the developing human fetal VM and functionally mature human DA neurons, which can be used to guide stem cell-based therapies and disease modeling approaches in PD.
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| 4. |
- Björklund, Anders, et al.
(författare)
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A Combined α-Synuclein/Fibril (SynFib) Model of Parkinson-Like Synucleinopathy Targeting the Nigrostriatal Dopamine System
- 2022
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Ingår i: Journal of Parkinson's Disease. - 1877-718X. ; 12:8, s. 2307-2320
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Forskningsöversikt (refereegranskat)abstract
- Injections of pre-formed α-synuclein fibrils (PFFs) or overexpression of α-synuclein using AAV vectors are commonly used as models of Parkinson-like synucleinopathy in rats and mice. In the modified method reviewed here, the "SynFib" model, the PFFs and the AAV vector are administered together unilaterally into the substantia nigra. This approach combines the key features of these two models, i.e., the generation of toxic α-synuclein aggregates and Lewy body-like inclusions, in combination with the increased vulnerability caused by increased cellular levels of α-synuclein. The combined AAV/PFF delivery offers several advantages over the standard PFF model due to the enhanced and accelerated α-synuclein pathology and microglial response induced by the PFF seeds in the presence of an elevated α-synuclein level. Injection of the AAV/PFF mixture into the substantia nigra makes it possible to target a larger proportion of the nigral dopamine neurons and obtain a level of dopamine cell loss (>60%) needed to induce significant impairments in drug-induced and spontaneous motor tests. The SynFib model shares attractive features of the standard 6-OHDA lesion model: a single unilateral stereotaxic intervention; pathology and cell loss developing over a short time span; and the possibility to monitor the degenerative changes using tests of motor behavior.
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| 5. |
- Butler, Simon, et al.
(författare)
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Considerations and challenges for the adoption of open source components in software-intensive businesses
- 2022
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Ingår i: Journal of Systems and Software. - : Elsevier. - 0164-1212 .- 1873-1228. ; 186
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Tidskriftsartikel (refereegranskat)abstract
- Component-Based Software Development is a conventional way of working for software-intensive businesses and OpenSource Software (OSS) components are frequently considered by businesses for adoption and inclusion in softwareproducts. Previous research has found a variety of practices used to support the adoption of OSS components, in-cluding formally specified processes and less formal, developer-led approaches, and that the practices used continue todevelop. Evolutionary pressures identified include the proliferation of available OSS components and increases in thepace of software development as businesses move towards continuous integration and delivery. We investigate workpractices used in six software-intensive businesses in the primary and secondary software sectors to understand currentapproaches to OSS component adoption and the challenges businesses face establishing effective work practices to eval-uate OSS components. We find businesses have established processes for evaluating OSS components and communitiesthat support more complex and nuanced considerations of the cost and risks of component adoption alongside matterssuch as licence compliance and functional requirements. We also found that the increasing pace and volume of softwaredevelopment within some businesses provides pressure to continue to evolve software evaluation processes.
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| 6. |
- Butler, Simon, et al.
(författare)
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On business adoption and use of reproducible builds for open and closed source software
- 2023
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Ingår i: Software quality journal. - : Springer Nature Switzerland AG. - 0963-9314 .- 1573-1367. ; 31:3, s. 687-719
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Tidskriftsartikel (refereegranskat)abstract
- Reproducible builds (R-Bs) are software engineering practices that reliably create bit-for-bit identical binary executable files from specified source code. R-Bs are applied in someopen source software (OSS) projects and distributions to allow verification that the distrib-uted binary has been built from the released source code. The use of R-Bs has been advo-cated in software maintenance and R-Bs are applied in the development of some OSS secu-rity applications. Nonetheless, industry application of R-Bs appears limited, and we seekto understand whether awareness is low or if significant technical and business reasonsprevent wider adoption. Through interviews with software practitioners and business man-agers, this study explores the utility of applying R-Bs in businesses in the primary and sec-ondary software sectors and the business and technical reasons supporting their adoption.We find businesses use R-Bs in the safety-critical and security domains, and R-Bs are valu-able for traceability and support collaborative software development. We also found thatR-Bs are valued as engineering processes and are seen as a badge of software quality, butwithout a tangible value proposition. There are good engineering reasons to use R-Bs inindustrial software development, and the principle of establishing correspondence betweensource code and binary offers opportunities for the development of further applications.
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| 7. |
- Fiorenzano, Alessandro, et al.
(författare)
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Single-cell transcriptomics captures features of human midbrain development and dopamine neuron diversity in brain organoids
- 2021
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Ingår i: Nature Communications. - : Springer Science and Business Media LLC. - 2041-1723. ; 12:1
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Tidskriftsartikel (refereegranskat)abstract
- Three-dimensional brain organoids have emerged as a valuable model system for studies of human brain development and pathology. Here we establish a midbrain organoid culture system to study the developmental trajectory from pluripotent stem cells to mature dopamine neurons. Using single cell RNA sequencing, we identify the presence of three molecularly distinct subtypes of human dopamine neurons with high similarity to those in developing and adult human midbrain. However, despite significant advancements in the field, the use of brain organoids can be limited by issues of reproducibility and incomplete maturation which was also observed in this study. We therefore designed bioengineered ventral midbrain organoids supported by recombinant spider-silk microfibers functionalized with full-length human laminin. We show that silk organoids reproduce key molecular aspects of dopamine neurogenesis and reduce inter-organoid variability in terms of cell type composition and dopamine neuron formation.
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| 8. |
- Giacomoni, Jessica, et al.
(författare)
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Protocol for optical clearing and imaging of fluorescently labeled ex vivo rat brain slices
- 2023
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Ingår i: STAR Protocols. - : Elsevier BV. - 2666-1667. ; 4:1
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Tidskriftsartikel (refereegranskat)abstract
- Tissue clearing is commonly used for whole-brain imaging but seldom used for brain slices. Here, we present a simple protocol to slice, immunostain, and clear sections of adult rat brains for subsequent high-resolution confocal imaging. The protocol does not require toxic reagents or specialized equipment. We also provide instructions for culturing of rat brain slices free floating on permeable culture inserts, maintained in regular CO2 incubators, and handled only at media change.
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| 9. |
- Hoban, Deirdre B, et al.
(författare)
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Impact of α-synuclein pathology on transplanted hESC-derived dopaminergic neurons in a humanized α-synuclein rat model of PD
- 2020
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Ingår i: Proceedings of the National Academy of Sciences of the United States of America. - : Proceedings of the National Academy of Sciences. - 1091-6490. ; 117:26, s. 15209-15220
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Tidskriftsartikel (refereegranskat)abstract
- Preclinical assessment of the therapeutic potential of dopamine (DA) neuron replacement in Parkinson's disease (PD) has primarily been performed in the 6-hydroxydopamine toxin model. While this is a good model to assess graft function, it does not reflect the pathological features or progressive nature of the disease. In this study, we establish a humanized transplantation model of PD that better recapitulates the main disease features, obtained by coinjection of preformed human α-synuclein (α-syn) fibrils and adeno-associated virus (AAV) expressing human wild-type α-syn unilaterally into the rat substantia nigra (SN). This model gives rise to DA neuron dysfunction and progressive loss of DA neurons from the SN and terminals in the striatum, accompanied by extensive α-syn pathology and a prominent inflammatory response, making it an interesting and relevant model in which to examine long-term function and integrity of transplanted neurons in a PD-like brain. We transplanted DA neurons derived from human embryonic stem cells (hESCs) into the striatum and assessed their survival, growth, and function over 6 to 18 wk. We show that the transplanted cells, even in the presence of ongoing pathology, are capable of innervating the DA-depleted striatum. However, on closer examination of the grafts, we found evidence of α-syn pathology in the form of inclusions of phosphorylated α-syn in a small fraction of the grafted DA neurons, indicating host-to-graft transfer of α-syn pathology, a phenomenon that has previously been observed in PD patients receiving fetal tissue grafts but has not been possible to demonstrate and study in toxin-based animal models.
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| 10. |
- Kirkeby, Agnete, et al.
(författare)
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Preclinical quality, safety, and efficacy of a human embryonic stem cell-derived product for the treatment of Parkinson's disease, STEM-PD
- 2023
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Ingår i: Cell Stem Cell. - 1934-5909. ; 30:10, s. 1299-1314
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Tidskriftsartikel (refereegranskat)abstract
- Cell replacement therapies for Parkinson's disease (PD) based on transplantation of pluripotent stem cell-derived dopaminergic neurons are now entering clinical trials. Here, we present quality, safety, and efficacy data supporting the first-in-human STEM-PD phase I/IIa clinical trial along with the trial design. The STEM-PD product was manufactured under GMP and quality tested in vitro and in vivo to meet regulatory requirements. Importantly, no adverse effects were observed upon testing of the product in a 39-week rat GLP safety study for toxicity, tumorigenicity, and biodistribution, and a non-GLP efficacy study confirmed that the transplanted cells mediated full functional recovery in a pre-clinical rat model of PD. We further observed highly comparable efficacy results between two different GMP batches, verifying that the product can be serially manufactured. A fully in vivo-tested batch of STEM-PD is now being used in a clinical trial of 8 patients with moderate PD, initiated in 2022.
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