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•Hh signaling shows diurnal oscillations in liver and hepatocytes in vitro and in vivo.•Hh signaling feeds-back on the liver clock via GLI transcription factors.•The amplitude of the ...oscillations of the liver clock is decreased in hepatocytes from Smo-knockout mice.•Rhythmicity of many metabolic pathways, including hepatic lipid metabolism, is affected by oscillating Hh signaling.•Diurnal timing of starvation affects the clock-hedgehog module differently.
The mammalian circadian clock controls various aspects of liver metabolism and integrates nutritional signals. Recently, we described Hedgehog (Hh) signaling as a novel regulator of liver lipid metabolism. Herein, we investigated crosstalk between hepatic Hh signaling and circadian rhythm.
Diurnal rhythms of Hh signaling were investigated in liver and hepatocytes from mice with ablation of Smoothened (SAC-KO) and crossbreeds with PER2::LUC reporter mice. By using genome-wide screening, qPCR, immunostaining, ELISA and RNAi experiments in vitro we identified relevant transcriptional regulatory steps. Shotgun lipidomics and metabolic cages were used for analysis of metabolic alterations and behavior.
Hh signaling showed diurnal oscillations in liver and hepatocytes in vitro. Correspondingly, the level of Indian Hh, oscillated in serum. Depletion of the clock gene Bmal1 in hepatocytes resulted in significant alterations in the expression of Hh genes. Conversely, SAC-KO mice showed altered expression of clock genes, confirmed by RNAi against Gli1 and Gli3. Genome-wide screening revealed that SAC-KO hepatocytes showed time-dependent alterations in various genes, particularly those associated with lipid metabolism. The clock/hedgehog module further plays a role in rhythmicity of steatosis, and in the response of the liver to a high-fat diet or to differently timed starvation.
For the first time, Hh signaling in hepatocytes was found to be time-of-day dependent and to feed back on the circadian clock. Our findings suggest an integrative role of Hh signaling, mediated mainly by GLI factors, in maintaining homeostasis of hepatic lipid metabolism by balancing the circadian clock.
The results of our investigation show for the first time that the Hh signaling in hepatocytes is time-of-day dependent, leading to differences not only in transcript levels but also in the amount of Hh ligands in peripheral blood. Conversely, Hh signaling is able to feed back to the circadian clock.
There have been some proposals that stem cells exist in the ovarian surface epithelium (OSE) of the adult human ovary; however, no direct evidence of such cells has been given until now. The aim of ...this study was to isolate the putative ovarian stem cells (OSCs) from the OSE layer in women with no naturally present oocytes and follicles—20 postmenopausal women and five women with premature ovarian failure. Small round cells with a bubble-like structure and diameters from 2 to 4 μm were isolated from the material obtained by OSE scraping. They expressed early embryonic developmental markers such as stage-specific embryonic antigen-4 and Oct-4, Nanog, Sox-2, and c-kit transcription markers, and they displayed prominent c-kit immunohistochemical staining. These cells were separated by density gradient centrifugation and grown
in vitro, where they proliferated. Some of them grew intensively and reached a diameter of approximately 20 μm after 5–7 days. In the OSE cell culture, oocyte-like cells developed, which reached a diameter of up to 95 μm and expressed Oct-4A, Oct-4B, c-kit, VASA, and ZP2 transcription markers, corresponding to early oocytes. They did not express SCP3 meiotic marker. In conclusion, the discovered cells are proposed to represent the adult OSCs with the expression of embryonic stem cell markers. The expression of germ lineage marker c-kit points toward their primordial germ cell ancestry. A new term “embryonic-like stem cells of the adult” is proposed for embryonic-like stem cells that might persist in various tissues and organs of adults. These findings could be used for further studies aimed at the autologous treatment of ovarian infertility and degenerative diseases.
Z dokumentom želimo smernice, ki jih predlaga GINA, vključiti v slovenski prostor. Želimo, da bi dokument služil enotnemu in dogovorjenemu pristopu k obravnavi bolnikov z astmo na primarni in ...specialistični pulmološki ravni.
Abstract Background: For better performance, it is crucial to continuously improve quality and patient safety based on the science of quality and patient safety and the usage of appropriate tools ...such as statistical process control. The aim of this article is to describe statistical process control in the continuous improvement of healthcare processes. Methods: Patient falls and surgical site infections were studied using a run chart, u-chart, and g-chart. Results: In the case of patient falls special causes of variation were depicted. However, these are only statistical conclusions, whereas the content of the causes can be found only by those who work in the process of fall prevention. In the case of surgical site infection, the use of a new wound covering material was abandoned because the g-chart showed failure to improve the process. The g-chart showed that a dierent type of preparation of the surgical site with hair cutting instead of shaving showed a decrease in infection rates. Conclusion: Statistical process control can differentiate between common and special causes of process variability. The importance of this distinction is in the employment of dierent strategies for process improvement.
Immunocytochemistry is a powerful tool for detection and visualization of specific molecules in living or fixed cells, their localization and their relative abundance. One of the most commonly used ...fluorescent DNA dyes in immunocytochemistry applications is 4′,6-diamidino-2-phenylindole dihydrochloride, known as DAPI. DAPI binds strongly to DNA and is used extensively for visualizing cell nuclei. It is excited by UV light and emits characteristic blue fluorescence. Here, we report a phenomenon based on an apparent photoconversion of DAPI that results in detection of a DAPI signal using a standard filter set for detection of green emission due to blue excitation. When a sample stained with DAPI only was first imaged with the green filter set (FITC/GFP), only a weak cytoplasmic autofluorescence was observed. Next, we imaged the sample with a DAPI filter set, obtaining a strong nuclear DAPI signal as expected. Upon reimaging the same samples with a FITC/GFP filter set, robust nuclear fluorescence was observed. We conclude that excitation with UV results in a photoconversion of DAPI that leads to detection of DAPI due to excitation and emission in the FITC/GFP channel. This phenomenon can affect data interpretation and lead to false-positive results when used together with fluorochrome-labeled nuclear proteins detected with blue excitation and green emission. In order to avoid misinterpretations, extra precaution should be taken to prepare staining solutions with low DAPI concentration and DAPI (UV excitation) images should be acquired after all other higher wavelength images. Of various DNA dyes tested, Hoechst 33342 exhibited the lowest photoconversion while that for DAPI and Hoechst 33258 was much stronger. Different fixation methods did not substantially affect the strength of photoconversion. We also suggest avoiding the use of mounting medium with high glycerol concentrations since glycerol showed the strongest impact on photoconversion. This photoconversion effect cannot be avoided even when using narrow bandpass filter sets.
Background: The sugar beet is the main field crop used for sugar production in the temperate climatic zone. The abolishment of the quota system will open new investment opportunities in countries ...that were forced to abandon sugar industry as the result of the reform in 2006. Present paper describes the modeling of sugar beet production and its processing into sugar for purpose of decision support.
Methods: A system dynamics methodology was chosen to model impacts of regional sugar factory investment. We present two basic concepts of system dynamics models at causal loop diagram level. The first holistic model deals with regional planning of new product development and the second one deals with factory model.
Results: The holistic model presented main feedback loops and dynamics of main elements in the case of regional investment into sugar industry. The factory model considered the specifics of the beet processing which is a) limited period of beet processing and b) initial adjustment to the production capacity at the start of the production season
Conclusions: The model seeks answers to strategic questions related to the whole sugar beet production and processing system and will be used for simulation of different scenarios for sugar production and their impact on economic and environmental parameters at an aggregate level.
We monitored trends of severe COVID-19 morbidity in Slovenia during weeks 13 to 37 2021. National weekly rates of severe acute respiratory infections (SARI) cases testing positive for SARS-CoV-2 at ...admission in all hospitals varied between 0.2 and 16.3 cases per 100,000 population. Of those without previous COVID-19 diagnosis, SARI COVID-19 admission rates ranged between 0.3 and 17.5 per 100,000 unvaccinated, and 0.0 and 7.3 per 100,000 fully vaccinated individuals. National SARI COVID-19 surveillance is essential in informing COVID-19 response.
In this paper, the model for organic farming development to support government decision making is presented. After performing several simulation scenarios, the authors discovered that conversion to ...organic farming relies on subsidies which provide the main motivation for conversion from conventional farming to organic farming. However, the subsidies are not the only driving force in the system, even more important are activities that promote organic farming, since sufficiently large subsidies could not be provided in order to complete the conversion from conventional to organic farming to the extent desired. Another important feature is food demand, determined by population size, which negatively influences the conversion to organic farming due to the increased demand leading to increased food prices.
Background. Glioblastoma multiforme (GBM) is a brain tumour with a very high patient mortality rate, with a median survival of 47 weeks. This might be improved by the identification of novel ...diagnostic, prognostic and predictive therapy-response biomarkers, preferentially through the monitoring of the patient blood. The aim of this study was to define the impact of GBM in terms of alterations of the plasma protein levels in these patients. Materials and methods. We used a commercially available antibody array that includes 656 antibodies to analyse blood plasma samples from 17 healthy volunteers in comparison with 17 blood plasma samples from patients with GBM.
Results. We identified 11 plasma proteins that are statistically most strongly associated with the presence of GBM. These proteins belong to three functional signalling pathways: T-cell signalling and immune responses; cell adhesion and migration; and cell-cycle control and apoptosis. Thus, we can consider this identified set of proteins as potential diagnostic biomarker candidates for GBM. In addition, a set of 16 plasma proteins were significantly associated with the overall survival of these patients with GBM. Guanine nucleotide binding protein alpha (GNAO1) was associated with both GBM presence and survival of patients with GBM.
Conclusions. Antibody array analysis represents a useful tool for the screening of plasma samples for potential cancer biomarker candidates in small-scale exploratory experiments; however, clinical validation of these candidates requires their further evaluation in a larger study on an independent cohort of patients.
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