101 research outputs found
deCLUTTER<sup>2+</sup> – a pipeline to analyze calcium traces in a stem cell model for ventral midbrain patterned astrocytes
Astrocytes are the most populous cell type of the human central nervous system and are essential for physiological brain function. Increasing evidence suggests multiple roles for astrocytes in Parkinson’s disease, nudging a shift in the research focus, which historically pivoted around ventral midbrain dopaminergic neurons (vmDANs). Studying human astrocytes and other cell types in vivo remains challenging. However, in vitro-reprogrammed human stem cell-based models provide a promising alternative. Here, we describe a novel protocol for astrocyte differentiation from human stem cell-derived vmDAN-generating progenitors. This protocol simulates the regionalization, gliogenic switch, radial migration and final differentiation that occur in the developing human brain. We characterized the morphological, molecular and functional features of these ventral midbrain patterned astrocytes with a broad palette of techniques and identified novel candidate midbrain-astrocyte specific markers. In addition, we developed a new pipeline for calcium imaging data analysis called deCLUTTER2+ (deconvolution of Ca2+ fluorescent patterns) that can be used to discover spontaneous or cue-dependent patterns of Ca2+ transients. Altogether, our protocol enables the characterization of the functional properties of human ventral midbrain patterned astrocytes under physiological conditions and in disease.</p
Decision-making based on 3D printed models in laparoscopic liver resections with intraoperative ultrasound: A prospective observational study
OBJECTIVES: The aim of this study was to evaluate impact of 3D printed models on decision-making in context of laparoscopic liver resections (LLR) performed with intraoperative ultrasound (IOUS) guidance.
METHODS: Nineteen patients with liver malignances (74% were colorectal cancer metastases) were prospectively qualified for LLR or radiofrequency ablation in a single center from April 2017 to December 2018. Models were 3DP in all cases based on CT and facilitated optical visualization of tumors\u27 relationships with portal and hepatic veins. Planned surgical extent and its changes were tracked after CT analysis and 3D model inspection, as well as intraoperatively using IOUS.
RESULTS: Nineteen patients were included in the analysis. Information from either 3DP or IOUS led to changes in the planned surgical approach in 13/19 (68%) patients. In 5/19 (26%) patients, the 3DP model altered the plan of the surgery preoperatively. In 4/19 (21%) patients, 3DP independently changed the approach. In one patient, IOUS modified the plan post-3DP. In 8/19 (42%) patients, 3DP model did not change the approach, whereas IOUS did. In total, IOUS altered surgical plans in 9 (47%) cases. Most of those changes (6/9; 67%) were caused by detection of additional lesions not visible on CT and 3DP.
CONCLUSIONS: 3DP can be helpful in planning complex and major LLRs and led to changes in surgical approach in 26.3% (5/19 patients) in our series. 3DP may serve as a useful adjunct to IOUS.
KEY POINTS: • 3D printing can help in decision-making before major and complex resections in patients with liver cancer. • In 5/19 patients, 3D printed model altered surgical plan preoperatively. • Most surgical plan changes based on intraoperative ultrasonography were caused by detection of additional lesions not visible on CT and 3D model
LRP10 and α-synuclein transmission in Lewy body diseases
Autosomal dominant variants in LRP10 have been identified in patients with Lewy body diseases (LBDs), including Parkinson's disease (PD), Parkinson's disease-dementia (PDD), and dementia with Lewy bodies (DLB). Nevertheless, there is little mechanistic insight into the role of LRP10 in disease pathogenesis. In the brains of control individuals, LRP10 is typically expressed in non-neuronal cells like astrocytes and neurovasculature, but in idiopathic and genetic cases of PD, PDD, and DLB, it is also present in α-synuclein-positive neuronal Lewy bodies. These observations raise the questions of what leads to the accumulation of LRP10 in Lewy bodies and whether a possible interaction between LRP10 and α-synuclein plays a role in disease pathogenesis. Here, we demonstrate that wild-type LRP10 is secreted via extracellular vesicles (EVs) and can be internalised via clathrin-dependent endocytosis. Additionally, we show that LRP10 secretion is highly sensitive to autophagy inhibition, which induces the formation of atypical LRP10 vesicular structures in neurons in human-induced pluripotent stem cells (iPSC)-derived brain organoids. Furthermore, we show that LRP10 overexpression leads to a strong induction of monomeric α-synuclein secretion, together with time-dependent, stress-sensitive changes in intracellular α-synuclein levels. Interestingly, patient-derived astrocytes carrying the c.1424 + 5G > A LRP10 variant secrete aberrant high-molecular-weight species of LRP10 in EV-free media fractions. Finally, we show that this truncated patient-derived LRP10 protein species (LRP10splice) binds to wild-type LRP10, reduces LRP10 wild-type levels, and antagonises the effect of LRP10 on α-synuclein levels and distribution. Together, this work provides initial evidence for a possible functional role of LRP10 in LBDs by modulating intra- and extracellular α-synuclein levels, and pathogenic mechanisms linked to the disease-associated c.1424 + 5G > A LRP10 variant, pointing towards potentially important disease mechanisms in LBDs.</p
LRP10 and α-synuclein transmission in Lewy body diseases
Autosomal dominant variants in LRP10 have been identified in patients with Lewy body diseases (LBDs), including Parkinson's disease (PD), Parkinson's disease-dementia (PDD), and dementia with Lewy bodies (DLB). Nevertheless, there is little mechanistic insight into the role of LRP10 in disease pathogenesis. In the brains of control individuals, LRP10 is typically expressed in non-neuronal cells like astrocytes and neurovasculature, but in idiopathic and genetic cases of PD, PDD, and DLB, it is also present in α-synuclein-positive neuronal Lewy bodies. These observations raise the questions of what leads to the accumulation of LRP10 in Lewy bodies and whether a possible interaction between LRP10 and α-synuclein plays a role in disease pathogenesis. Here, we demonstrate that wild-type LRP10 is secreted via extracellular vesicles (EVs) and can be internalised via clathrin-dependent endocytosis. Additionally, we show that LRP10 secretion is highly sensitive to autophagy inhibition, which induces the formation of atypical LRP10 vesicular structures in neurons in human-induced pluripotent stem cells (iPSC)-derived brain organoids. Furthermore, we show that LRP10 overexpression leads to a strong induction of monomeric α-synuclein secretion, together with time-dependent, stress-sensitive changes in intracellular α-synuclein levels. Interestingly, patient-derived astrocytes carrying the c.1424 + 5G > A LRP10 variant secrete aberrant high-molecular-weight species of LRP10 in EV-free media fractions. Finally, we show that this truncated patient-derived LRP10 protein species (LRP10splice) binds to wild-type LRP10, reduces LRP10 wild-type levels, and antagonises the effect of LRP10 on α-synuclein levels and distribution. Together, this work provides initial evidence for a possible functional role of LRP10 in LBDs by modulating intra- and extracellular α-synuclein levels, and pathogenic mechanisms linked to the disease-associated c.1424 + 5G > A LRP10 variant, pointing towards potentially important disease mechanisms in LBDs.</p
Co-creation of a virtual interactive teaching package for auditors of healthcare placements - towards assurance of quality of health care traineeships
To provide medical and allied health professionals students with the best clinical learning environments, quality processes must be in place, and these require innovation to assure audit material resources that are fit for purpose, can work well within the situation and provide the correct teaching and learning to train auditors. HEALINT4ALL ERASMUS+ Strategic partnership provides medical education and allied health professionals with an audit system to facilitate quality assurance of EU clinical learning environments by mapping and innovatively adapting a newly established audit protocol and support tools to suit the Higher Education needs for wider application to medicine and professionals allied to medicine. The project conducted a literature scoping review followed by interviews and focus groups across all 6 European partners of clinicians, students and educators of service needs and best practice, resulting to map standards and requirements for clinical learning environments and develop a protocol to assess the quality of placements. Next, it developed a digital interactive platform for European and national placements appraisal following user-centred design to allow the collaboration between Higher Education Institutions (HEIs), and HEIs and placements. In order the application of the HEALINT4ALL protocol within the digital interactive tool to be beneficial, the auditors of the healthcare placements should have the necessary competences to perform the audits utilising the HEALINT4ALL a digital interactive platform. An online or blended learning approach is preferred to fit with the clinical academics' needs that will undertake the role of auditor. Thus, HEALINT4ALL co-created a Virtual Interactive Teaching Package for Training the Auditors of Healthcare Placements. It followed a modified ASPIRE framework to develop the package. The ASPIRE framework stands for Aims, Storyboarding, Population, Implementation, Release and Evaluation. This work describes the co-creation journey that engaged stakeholders in 6 different countries in order to co-design the resources, experts in the field that reviewed the content to ensure its high quality, the development that followed and the first pilot's evaluation from experts. While some efforts have been made towards the standardisation of auditing clinical placements, to the best of our knowledge this is the first attempt to develop a short curriculum to training the auditors of healthcare placements and implemented as a virtual interactive teaching package
Metabolic and endocrine profiles and reproductive parameters in dairy cows under grazing conditions: effect of polymorphisms in somatotropic axis genes
<p>Abstract</p> <p>Background</p> <p>The present study hypothesized that GH-AluI and IGF-I-SnabI polymorphisms do change the metabolic/endocrine profiles in Holstein cows during the transition period, which in turn are associated with productive and reproductive parameters.</p> <p>Methods</p> <p>Holstein cows (Farm 1, primiparous cows, n = 110, and Farm 2, multiparous cows, n = 76) under grazing conditions were selected and GH and IGF-I genotypes were determined. Blood samples for metabolic/endocrine determinations were taken during the transition period and early lactation in both farms. Data was analyzed by farm using a repeated measures analyses including GH and IGF-I genotypes, days and interactions as fixed effects, sire and cow as random effects and calving date as covariate.</p> <p>Results and Discussion</p> <p>Frequencies of GH and IGF-I alleles were L:0.84, V:0.16 and A:0.60, B:0.40, respectively. The GH genotype was not associated with productive or reproductive variables, but interaction with days affected FCM yield in multiparous (farm 2) cows (LL yielded more than LV cows) in early lactation. The GH genotype affected NEFA and IGF-I concentrations in farm 1 (LV had higher NEFA and lower IGF-I than LL cows) suggesting a better energy status of LL cows.</p> <p>There was no effect of IGF-I genotype on productive variables, but a trend was found for FCM in farm 2 (AB cows yielded more than AA cows). IGF-I genotype affected calving first service interval in farm 1, and the interaction with days tended to affect FCM yield (AB cows had a shorter interval and yielded more FCM than BB cows). IGF-I genotype affected BHB, NEFA, and insulin concentrations in farm 1: primiparous BB cows had lower NEFA and BHB and higher insulin concentrations. In farm 2, there was no effect of IGF-I genotype, but there was an interaction with days on IGF-I concentration, suggesting a greater uncoupling somatropic axis in AB and BB than AA cows, being in accordance with greater FCM yield in AB cows.</p> <p>Conclusion</p> <p>The GH and IGF-I genotypes had no substantial effect on productive parameters, although IGF-I genotype affected calving-first service interval in primiparous cows. Besides, these genotypes may modify the endocrine/metabolic profiles of the transition dairy cow under grazing conditions.</p
The surface runoff of calcium, magnesium, iron, manganese, nitrogen and phosphorus from the upper Pasłęka river catchment
Celem badań było ustalenie wartości jednostkowych współczynników spływu makropierwiastków z terenów o różnych formach zagospodarowania w zlewni górnej Pasłęki i jej prawobrzeżnego dopływu Giłwy. Stwierdzono, że największe wartości współczynników spływu manganu oraz azotu i fosforu do wód uzyskano z obszarów użytkowanych rolniczo (110,8 kg Ca∙ha-1∙rok-1, 15,6 kg Mg∙ha-1 ∙rok-1, 0,5 kg Mn∙ha-1∙rok-1, 0,6 kg P∙ha-1∙rok-1, 14,3 kg N∙ha-1∙rok -1). Badania wykazały też wpływ terenów użytkowanych rolniczo, zarówno gruntów ornych, jak i użytków zielonych (łąki i pastwiska), na zwiększenie ładunku obszarowego wapnia i magnezu, w odniesieniu do których uzyskano największe współczynniki spływu z tych form użytkowania ziemi. Największą wartość współczynników spływu żelaza uzyskano dla obszarów leśnych (0,6 kg Fe∙ha-1 ∙rok-1).The aim of study was to determine values of unit coefficients of the surface runoff of macroelements from surfaces with different land management types, which was conducted in the catchment basin of the upper Pasłęka River and its right-bank tributary, the Giłwa River. The highest values of the coefficients describing the runoff of manganese as well as nitrogen and phosphorus to waters were obtained for a subcatchment comprising farmland (110.8 kg Ca∙ha-1·year-1, 15.6 kg Mg·ha-1·year-1, 0.5 kg Mn·ha-1·year-1, 0.6 kg P·ha-1·year-1, 14.3 kg N·ha-1·year-1 ). The study also pointed up the effect of agriculturally used areas, both arable land and grassland (meadows and pastures), consisting in increasing the non-point loads of calcium and magnesium, which scored the highest runoff coefficient values for the mentioned types of land use. The highest value of the iron runoff coefficient was calculated for forested areas (0.6 kg Fe·ha-1·year-1)
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