1. I ett samarbetsprojekt har 14 forskare från Frankrike,Kina och Australien fått sina studier publicerade om ett ämne jag inte är närmare bekant med,nämligen hydrogeler inom Reg Med som material att använda vid 3D vävnadstillverkning.
Abstract
Currently, there are only few reports on water-soluble photoinitiating systems. In this study, a highly water-soluble organic dye i.e. sodium (E)-3,3′-((4-(2-(3-methylbenzo[d]thiazol-3-ium-2-yl)vinyl)phenyl)azanediyl)dipropionate iodide, was synthesized and served as a photoinitiator. Notably, this water-soluble initiator, at a low concentration of just 0.01 wt%, demonstrates a high photoinitiation ability, with some hydrogel formulations achieving nearly 100% double bond conversion under sunlight. Photopolymerization kinetics were monitored using Real-Time Fourier Transform Infrared. To explore the complex chemical principles of radical polymerization, UV-visible absorption and fluorescence spectroscopy, steady-state photolysis, fluorescence quenching experiments and cyclic voltammetry were employed to gain a comprehensive understanding of the photochemical mechanism involved. Additionally, several characteristics of the synthesized hydrogels were also investigated i.e. the water content, the water swelling, and the volume swelling. In addition to their excellent photoinitiation capabilities, the hydrogel formulations developed in this study also supported 3D printing. 3D objects with smooth surface and a high spatial resolution could be successfully printed using direct laser writing. The fabricated hydrogels could reversibly change of shape in response to water (adding or removing water), enabling successful 4D printing behavior. Furthermore, the efficient photoinitiation ability of the water-soluble formulations opens new avenues for sunlight-polymerized hydrogels and potential applications in bioprinting.
2.2 Study on Cytotoxicity of Dye-A1
It can be seen that the cells have already died, with only few cells remaining visible under the Holomonitor (see Figure S2, Supporting Information). As shown in Figure S3 (Supporting Information), the track of single cell movement in the culture medium containing dye-A1 shows that the cells had vitality, and the number of cells was significantly higher than that of the control TPO (see Figure S4, Supporting Information). The above results show that dye-A1 did not cause any cytotoxicity. Therefore, dye-A1 can support the growth, proliferation, and survival of cells, demonstrating excellent biocompatibility.
|
Figure S4. The Holomonitor images of dye-A1. |
--------------------------------------------------------------------------------
2. 6 forskare från Taiwan har fått sina studier om neutrofila granulocyter,som är vita blodkroppar och ingår i kroppens immunsystem,publicerade. Studien är dessvärre hårt inlåst men Google Scholar har lagt den under Studier med HoloMonitor så vi får lita på dem. Chang Gung är ett HoloMonitoruniversitet så det vidimerar påståendet.
--------------------------------------------------------------------------------
3. 4 Forskare från Grekland har fått sina studier om probiotika (mölksyrebakterier) kontra celldöd publicerade. Noterbart att Grekland inte finns upptagna under PHI`s User-lista så det bör vara en ny kund.
Lactobacilli-host interactions inhibit Staphylococcus aureus and Escherichia coli-induced cell death and invasion in a cellular model of infection 18 Dec 2024Probiotic lactobacilli exert antimicrobial and antibiofilm activity through a plethora of mechanisms, including the production of inhibitory compounds and competition for nutrients and adhesion sites on the host mucosa. Their ability to interfere with pathogen-host interactions may mitigate the harmful effects of infection. Lactiplantibacillus pentosus L33 (L33) and Lactiplantibacillus plantarum L125 (L125) are two potential probiotic lactic acid bacteria (LAB) strains, previously shown to reduce pathogen viability and biofilm formation in vitro. The present study aims to investigate their ability to limit cell death induced by Staphylococcus aureus and Escherichia coli in the human colon adenocarcinoma cell line HT-29. To this end, we examined the protective effects of the two strains using in silico, in vitro and omic approaches, with Lacticaseibacillus rhamnosus GG (LGG) serving as a reference strain, due to its well-documented antimicrobial properties. Based on the findings of our study, direct contact of HT-29 cells with L125 for 4 h prior to the addition of S. aureus or E. coli prevented pathogen-induced cell death at rates comparable to LGG. In contrast, L33 failed to exert a protective effect. Moreover, L125 significantly reduced adherence of S. aureus to HT-29 cells, and the internalization capacity of both pathogens (>1.5 Log CFU/mL). Dual RNA-seq and protein microarrays were used to determine expression changes in L125 and host cells during co-incubation. L125 expressed high levels of adhesins and moonlighting proteins, homologous to those encoded by the pathogens. Pathways involved in pathogen adhesion and internalization, endocytosis, cell–cell and cell-extracellular matrix (ECM) adhesion, were downregulated in HT-29 cells. Finally, L125 reduced the secretion of various pro-inflammatory mediators. Our findings highlight the strain-specific protective effects of LAB against pathogen-induced cell death achieved through competitive exclusion and priming of host cell responses. Future studies will focus on elucidating the specific surface components of L125 involved in these events, paving the way for targeted interventions at the host-pathogen interface.
2.3 Cell and bacteria viability assays
The cytotoxic capacity of pathogen CM was also evaluated using the HoloMonitor Live Imaging System (PHI, Boston, MA, United States). In this case, 2 × 105 cells per well were seeded into 24-well plates. The following day, cells were treated with undiluted, sterile-filtered pathogen CM. Cells treated with cell culture medium were included as an untreated control. Cell proliferation was assessed over a 24 h period with the Kinetic Cell Proliferation Assay software (PHI).
3.1 Staphylococcus aureus and Escherichia coli induce cell death of HT-29 cells in a time-dependent manner
These effects were further monitored for 24 h with the HoloMonitor live cell imaging system, using the Kinetic Cell Proliferation Assay software. It was shown that CM derived from either pathogen reduced cell confluency in a time dependent manner (Figure 1F).
|
(F) The effect of pathogen-derived CM was monitored for 24 h using the HoloMonitor live cell imaging system. SA CM and EC CM decreased confluency in HT-29 cells in a time-dependent manner. The data presented are the mean ± standard deviation of three independent experiments. *p < 0.05 and **p < 0.005 compared to control (CTRL) untreated cells. |
--------------------------------------------------------------------------------4. 22 kinesiska forskare har fått sina studier om elakartade cancertumörer har haft påverkan av ett specifikt kroppseget protein (SAMHD1) och om så hitta motmedel.
Nuclear Overexpression of SAMHD1 Induces M Phase Stalling in Hepatoma Cells and Suppresses HCC Progression by Interacting with the Cohesin Complex
Emerging evidence suggests that the sterile alpha-motif (SAM) and histidine
aspartate (HD)domain-containingprotein1(SAMHD1)isimplicatedinvarious
cancers, including hepatocellular carcinoma (HCC). However, its precise role in
tumor cells and the underlying mechanisms remain unclear. This study aimed
to investigate the expression patterns, prognostic values, and functional role of
SAMHD1inHCCprogression. We constructed liver tissue microarrays using
tumor and paired paratumor tissue specimens from 187 patients with primary
HCC. Our findings indicate that nuclear SAMHD1 protein levels are increased
in tumors compared to paratumor tissues. Moreover, nuclear SAMHD1
levels decline in advanced tumor stages, with higher SAMHD1 nuclear
staining correlating with favorable prognostic outcomes. Hepatocyte-specific
SAMHD1knockout mice, generated by crossing SAMHD1fl/fl mice with
Alb-cre mice, showed accelerated tumor progression in a diethylnitrosamine
(DEN)-induced HCC model. In hepatoma cell lines, nuclear
overexpression of SAMHD1 inhibited cell proliferation by stalling mitosis,
independent of its deoxynucleotide triphosphohydrolase (dNTPase) function.
Mechanistically, SAMHD1 interacts with the cohesin complex in nucleus,
enhancing sister chromatid cohesion during cell division, which delays
metaphase progression. Our findings suggest that nuclear SAMHD1 plays
a critical role in slowing HCC progression by regulating mitosis, highligh
Cell migration was evaluated using a wound healing assay and a Tran
swell assay. For the wound healing assay, cells were seeded in 6-well plates
and grown to confluence. A scratch was made with a sterile pipette tip,
followed by washing to remove detached cells. Wound closure was mon
itored over time using phase-contrast microscopy, and images were ana
lyzed with ImageJ to calculate the percentage of wound closure relative to
the initial area. For the Transwell assay, cells were resuspended in serum
free DMEM at a density of 7.5 × 104 cells mL−1. A total of 200 μL of this
suspension was added to the upper chamber, with 500 μL of DMEM con
taining 20% FBS in the lower chamber. After 24 h of incubation, migrated
cells were fixed with 4% paraformaldehyde and stained with 0.1% crystal
violet.
To assess the impact of cohesion complex knockdown on the pro
liferation and migration of SAMHD1-overexpressing HepG2 cells, cells
were suspended in a medium containing a mixture of RNAiMax transfec
tion reagent with scrambled siRNA or RAD21 and SMC3 siRNA mixture
and were seeded in 6-well plates at 20% confluence for 24 h. Then the
cell plates were placed into the HoloMonitor M4 digital holographic mi
croscopy (Phase Holographic Imaging PHI Inc) within a standard CO2 in
cubator. Six regions of interest (ROI) in each well were randomly selected
by the instrument software, and images were captured every 2 h for a con
tinuous 52-h period.
The HoloMonitor software’s Kinetic Cell Proliferation
Assay and Kinetic Cell Motility Assay modules were used to quantify cell
growth and calculate average migration speed based on the captured im
ages.
--------------------------------------------------------------------------------
5. 9 forskare från kända John Hopkins University USA har genomfört riktigt avancerade studier på celler och deras förmåga till anpassning? vid yttre påverkan. Forskning på allra högsta nivå känns adekvat för en lekman som jag att säga. Bedöm själva genom att läsa hela studien. Och av intresse: De pratar om QPM som teknik (Quantative Phase Microscopy) de använt för sina resultat. Med det menat att de använt HoloMonitor som vi kanske bättre känner igen som QPI. Närmare bestämt HoloMonitor tillsammans med Single Cell assay:en.
I Introduction
How mammalian cells coordinate growth and maintain size across changing environments is a fundamental question in cell biology.
Recent advances in single-cell methods have enabled precise measurements of cell volume, mass, and cell mass density (CMD). For instance, microfluidic-based methods such as the Fluorescence eXclusion method (FXm) can accurately measure cell volume in the 10-100 femto-liter range.Micro-cantilever methods have been employed to measure cell dry mass in the femto-gram (fg) range.
Optical methods such as the Quantitative Phase Microscopy (QPM) can also accurately determine the dry mass (without water) of single cells with pico-gram sensitivity.Therefore, an unprecedented window is opening for examining live cell mass and volume, and also the cell mass density (mass/volume). In this paper, utilizing a combination of FXm and QPM, we quantitatively explore CMD regulation after media osmolarity change. We discover that while complex changes occur in cell volume and mass after osmotic shock, cells prioritize CMD recovery in a straightforward manner, suggesting that there is a simple control algorithm of CMD.
Quantitative phase microscopy
Cell dry mass represents the total mass of all intracellular non-water molecules. To measure cell dry mass, we utilized the Holomonitor M4 quantitative phase microscope (PHI). Quantitative phase microscopy (QPM) is based on the principle that the refractive index of a material inversely correlates with the velocity of light passing through it. When in operation, the Holomonitor directs a laser beam through both the cell and the surrounding culture medium. As light passes through the cell, which has a higher refractive index than the medium, it slows down, resulting in a detectable phase shift. This phase shift is captured by the camera and processed to summarize the total phase alteration caused by the entire cell (optical volume). The total phase alteration of individual cells was analyzed using the dedicated Holomonitor M4 software. For aqueous solutions, the refractive index is linearly proportional to the solute mass concentration, with a consistent factor α = 0.18 mL/g irrespective of the solute type.We have also validated this factor using a NaCl and a BSA solution. After obtaining the cell volume, the optical volume, and culture medium refractive index, cell mass can be calculated as:where Δφ is the phase shift cells make in each pixel, nm is refractive index of cell culture medium, nwater is refractive index of water. The refractive index of different media was measured using the Digital Brix Refractometer (MILWAUKEE Instruments MA871).
--------------------------------------------------------------------------------
6. Som kronan på verket kommer slutligen en tvättäkta HoloMonitorbaserad studie där 7 polska forskare verkligen gått på djupet med genstudier (miR-1244) kopplat till cancer. Hänvisningarna till HoloMonitor är för många att ta upp så det blir ett urval. Studien är publicerad hos det förnämsta vetenskapliga organet av de alla : Springer Nature
The Yin and Yang of hsa-miR-1244 expression levels during activation of the UPR control cell fate
Abstract
Regulation of endoplasmic reticulum (ER) homeostasis plays a critical role in maintaining cell survival. When ER stress occurs, a network of three pathways called the unfolded protein response (UPR) is activated to reestablish homeostasis. While it is known that there is cross-talk between these pathways, how this complex network is regulated is not entirely clear. Using human cancer and non-cancer cell lines, two different genome-wide approaches, and two different ER stress models, we searched for miRNAs that were decreased during the UPR and surprisingly found only one, miR-1244, that was found under all these conditions. We also verified that ER-stress related downregulation of miR-1244 expression occurred with 5 different ER stressors and was confirmed in another human cell line (HeLa S3). These analyses demonstrated that the outcome of this reduction during ER stress supported both IRE1 signaling and elevated BIP expression. Further analysis using inhibitors specific for IRE1, ATF6, and PERK also revealed that this miRNA is impacted by all three pathways of the UPR. This is the first example of a complex mechanism by which this miRNA serves as a regulatory check point for all 3 pathways that is switched off during UPR activation. In summary, the results indicate that ER stress reduction of miR-1244 expression contributes to the pro-survival arm of UPR.
Real-time cell viability assay
For real-time monitoring of cell viability, we applied real-time and label-free holographic microscopy-based monitoring of cell death and viability using HoloMonitor M4® time-lapse cytometer (Phase Holographic Imaging PHI AB, Lund, Sweden).
Holographic microscopy was used to follow the optical thickness and irregularity of cells exposed for up to 24 h to Tm or Tg in the presence or absence of miR-1244 mimic or antagomiR. The images from up to 8 independent optical fields were collected and analyzed according to manufactures instructions with HoloMonitor® App Suite software. Healthy cells are irregular in shape and thin, whereas dying cells are round and thick. For all analysis, the same cells parameters qualification was applied.
To better understand this the cell fate decision dynamics, we performed real time and label free holographic microscopy-based monitoring of cell death and viability using a
HoloMonitor® time-lapse cytometer. Holographic microscopy was used to follow the optical thickness and irregularity of cells exposed for up to 24 h to Tm and Tg in the presence or absence of miR-1244 mimic or antagomiR (Figs.
4,
5 and
6 and Supplemental Figs.
6–
8).
Exogenous miR-1244 influences the fate of cells challenged with tg induced ER stress. The results of real-time monitoring of cell viability with the real time and label free holographic microscopy are shown using a HoloMonitor M4® time-lapse cytometer of HeLa cells transfected with miR-1244 mimic or inhibitor or the scramble control and 48 h later treated with Tg (500 nM) up to 24 h. Images were collected every 15 min (from 5 independent optical fields), and the distribution of live (blue) and dying cells (red) based on their optical thickness (Y-axis) and irregularity (X-axis) is presented at the 0, 8, 16 and 24 h time points. The images from up to 5 independent optical fields were collected and analyzed according to manufacturer’s instructions with HoloMonitor® App Suite software.
Genstudier med HoloMonitor !
Min kommentar
HoloMonitor avslutar detta år med 6 forskningsrapporter där 2 är extra vassa och förmodligen kommer användas i PHI`s marknadsföring. Ett år som för oss aktieägare inte varit nån höjdare ska ärligt sägas.
Men instrumentet skördar nya segrar som synes och det vore väl fasiken om inte vår nya ledning ska kunna kapitalisera på det konstaterandet. 2025 ska bli året då PHI och aktien gör en hejdundrande come back har jag bestämt. 😎
Mvh the99