NK cell activity and T4 excretions increased in the fibrotic group with NPA tumor induction.

<p>(A) Intra-hepatic NK cells as well as (B) NK from spleen from four groups were isolated and stained for NK activity (CD107a) using flow-cytometry. The data showed a significant activation of the intrahepatic and splenic NK from the fibrotic group with NPA tumor induction compared to the non-tumor injected animals. (C) Shows serum T4 levels significantly elevated in animal models of fibrosis following NPA tumor injections.</p

Tumor increase severity of hepatic fibrosis.

<p>Fibrotic profile was estimated by (A) Western blot quantitations and (B) RT-PCR expressions of α smooth muscle actin (αSMA). (A) (Lower panel) displays a representative membrane with examples of αSMA expression as a marker for the HSCs activation (upper bands) and GAPDH (lower bands) expression by western blotting in the harvested liver protein extracts. (A) (Upper panel) shows the calculated ratio of αSMA/β-actin based on the densitometry readings of the bands. (B) Real-time PCR data reflect changes in gene expression of αSMA mRNA expressed as fold change compared with naïve mice. Expression of αSMA mRNA corresponded to the western blot results. Experiments were repeated 4 times, in each time 4 mice were included in each group. Averages of the 4 experiments were included in the quantitations of western blots and RT PCR.</p

VEGF serum levels.

<p>Quantikine Mouse VEGF immunoassay serum levels was significantly increased in the fibrotic group bearing tumor as compared to tumor alone (p = 0.01) or fibrosis alone groups (p = 0.04). No statistical significant differences were found in VEGF serum levels between the fibrotic and non-fibrotic group with tumor induction.</p

In vitro co-culture of lymphocytes with NPA cell line.

<p>Adhered-NPA cells post co-culture with NK cells from different animal groups were analyzed for proliferation by CFSE using flow cytometry. (A) Direct co-culture of NPA cells with spleen NKs from fibrotic mice with tumor significantly decreased NPA tumor cell proliferation compared to the fibrotic mice without tumor, indicating highly stimulated NK cells effects; p-value = 0.001. (B) A representative histogram of the NPA cells following incubations with NK cells of fibrosis and tumor mice. The histogram shows CSFE-proliferations changes in day 3 and day 5 as compared to day 0 of CSFE staining. Proliferations fold changes were calculated by divided day 0 to day 5.</p

NPA Tumor inductions increase the severity of CCl4 related hepatic injury.

<p>CCl4-hepatic injury was evaluated by hematoxylin and eosin (H&E) staining of necro-inflammatory liver lesions and ALT serum levels. Immunohistochemical staining with H&E (5X magnification) for the four major animal groups showed necro-inflammatory lesions and cell infiltrations that were increased in the fibrotic mice receiving the NPA-tumor cells (D) as compared to fibrotic alone (C). Arrows indicate the area with lymphocyte infiltrations. No inflammatory infiltrates were seen in H&E staining of (A) naïve WT and (B) naïve mice receiving the NPA-tumor cells. (E) Serum ALT levels were in line with histological findings and showed increase from (60±25/L) in fibrotic animals without tumor to (85.5 ± 20.5 U/L) in animals with tumor and hepatic fibrosis; p-value = 0.021.</p

Hepatic fibrosis increase NPA tumor weight and size.

<p>In vivo S.C injection of NPA cells model was performed as described in M&M. In this model; S.C NPA-cells tumor was induced in fibrotic and naïve (no fibrosis) animals. S.C tumors were explanted at the end of 6 weeks post S.C injection, and evaluated for tumor weight and volume. (A) and (B) show the external appearance of the tumor in the animal’s back. Tumor weight (C) and volume (D) was increased significantly from (0.13±0.06gr) and (0.28±0.18ml) in the fibrotic group to (0.05±0.025 gr) and (0.09±0.01ml) in the non-fibrotic group; p-value = 0.02 and 0.04, respectively.</p

Sinusoidal capillarization in sVEGF-R1 expressing livers without parenchymal damage.

<p>(a,b) Immunohistochemical staining for vWF (Von-Willebrand Factor) on liver sections (black arrows-sinusoids, white arrows-larger blood vessels). (c) H&E staining of liver sections showing normal appearance.</p

Image_1_Patients with low ALT levels are at increased risk for severe COVID-19.jpg

IntroductionFrailty is a known risk factor for many diseases, including COVID-19. However, many frail patients are undiagnosed as the diagnosis can be cumbersome. Alanine transaminase (ALT) is found not only in the liver but also in the muscle tissue, and multiple studies show that frail sarcopenic patients have lower ALT. Frail patients are at increased risk for severe COVID-19. We evaluated the association between pre-infection low ALT and the risk for severe COVID-19.MethodsWe collected data regarding all subjects tested for SARS-CoV-2 between 1 March 2020 and 31 December 2021 from a national state-mandatory HMO in Israel, serving more than 1.3 million patients. Clinical and laboratory data were collected, including ALT from the year prior to infection. Severe COVID-19 was defined either as death, ICU admission, or ≥10 hospitalization days. Patients with low ALT (ALT ≤ 10 IU/l) were compared with patients with normal ALT (11–40 IU/l). Patients younger than 18 years with a diagnosis of liver disease and with ALT > 40 IU/l were excluded.ResultsDuring the study period, 58,961 patients tested positive for SARS-CoV-2. The patients in the low ALT group were younger (40.53 vs. 42.73, p ConclusionLow ALT level prior to infection is a significant risk factor for morbidity and mortality from COVID-19 infection. Further studies are warranted to address treatment options for this population.</p

sVEGF-R1 expression in the adult liver causes activation of HSCs.

<p>(a) Scanning electron microscopy of sinusoids in ‘off’ (a control littermate) vs. switch ‘on’ liver for one month showing prominent HSCs surrounding sinusoids (arrows) (b) Scanning electron microscopy showing transformation of HSCs from lipid droplets (L) containing cells (‘off’) to myofibroblasts like cells (arrows). (c) Quantification of the surface area of HSCs. ‘off’-5.8%, ‘on’ (sVEGF-R1 expression for one month)-43.7%. (d) Western blot analysis with anti α-smooth muscle actin antibody performed on liver extracts (e) Goldner staining for collagen fibers (green) indicating perisinusoidal accumulation of extra-cellular matrix.</p

sVEGF-R1 expression in the adult liver causes closure of sinusoidal fenestrations.

<p>(a) Scanning electron microscopy of sinusoids in control (‘off’) showing fenestrations arranged in sieve plates (white arrows) and loss of fenestration following one month of switch ‘on’ (sVEGF-R1 expression). RBC = Red Blood Cell (b) Quantification of the percent of fenestrations' surface area in the sinusoids. sinusoidal area. ‘off’−40.5%, ‘on’−8.5%.</p