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PubMed2026

Discovery of NAT-6-321056 as a novel modulator of VEGFR2 signaling to suppress tumor angiogenesis.

Vascular endothelial growth factor receptor 2 (VEGFR2) is a master regulator of angiogenesis and cancer progression. However, current VEGFR2 modulators face significant challenges, including off-target toxicity and acquired resistance, underscoring the urgent need for novel therapeutic agents with improved efficacy and safety profiles. Here, we reported that virtual screening of 39,442 natural products from the ZINC natural products-derived library, coupled with molecular docking and molecular dynamics (MD) simulations to evaluate the binding stability of candidate compounds, identified NAT-6-321056 as a highly promising modulator of VEGFR2 signaling. Biological evaluations demonstrated that NAT-6-321056 exerted potent inhibition on the growth of a broad spectrum of cancer cells, including both solid tumors and hematological malignancies. In EA.hy 926 endothelial cells and SK-N-DZ neuroblast cells, the compound significantly suppressed proliferation, migration, and invasion. Microscale thermophoresis (MST) confirmed direct binding of NAT-6-321056 to VEGFR2 with favorable affinity. Kinase profiling against a panel of 33 kinases indicated that NAT-6-321056 exhibited a multi-kinase modulation profile. Mechanistic studies revealed that NAT-6-321056 suppressed the expression of hypoxia-inducible factor 1-alpha (HIF-1α) and was associated with reduced VEGFR2 phosphorylation and attenuation of the downstream ERK/JNK/AKT signaling pathways. Moreover, NAT-6-321056 exhibited robust in vivo anti-angiogenic effects in both the chick chorioallantoic membrane (CAM) assay and transgenic zebrafish vascular fluorescence imaging models. Computational absorption, distribution, metabolism, excretion, and toxicity (ADMET) prediction suggested acceptable drug-like properties. Collectively, these findings demonstrated that NAT-6-321056 is a promising modulator of VEGFR2 signaling with potent anti-angiogenic activity and represents a viable candidate for cancer therapy.

Vascular Endothelial Growth Factor Receptor-2↗

PubMed1975

In ovo assay for Marek's disease virus and turkey herpesvirus.

Marek's disease virus (MDV) and the turkey herpesvirus (HVT) may be assayed on the chorioallantoic membrane (CAM) of the chicken embryo after intravenous inoculation of chicken embryo fibroblasts (CEF) or chicken blood leukocytes infected with these viruses. Free HVT, MDV associated with Marek's tumor cells, and lymphoblastoid cell lines derived from Marek's tumors, may be assayed in the same way. The intravenous assay is quicker than the yolk sac assay and somewhat more sensitive than in vitro or conventional CAM assay after direct inoculation of the CAM. The optimal time for inoculation was day 10 of embryo incubation; therafter the log-10 CAM lesions decreased as a negative linear function of embryo age at the time of inoculation. The log-10 CAM lesions increased as a positive linear function of the time since inoculation. The optimal time for counts was day 5 after inoculation. The log-10 CAM lesions was a linear function of the log-10 cells in the inoculum; the slope was 1.0. Venous in ovo inoculation caused as increase in the weight of the spleen proportional to the number of CAM lesions. Repression of the splenomegaly, by prior X irradiation of the embryo, did not reduce the number of CAM lesions. Embryols from lines inbred for susceptibility to Marek's disease produced more CAM lesions than embryos from resistant lines. This difference did not depend on prior exposure of the mothers to MDV or HVT.

Allantois↗

PubMed1977

Human somatotropin 53. Synthesis and biological activity of the amino terminal fifty-four residue fragment.

The amino terminal 54 residue peptide fragment of human somatotropin [Cys(Gam)53]-HGH-(1-54), has been synthesized by the solid-phase method. The symmetrical anhydride and active ester coupling methods were used exclusively. The synthetic product was purified by gel fitration isoelectric focusing, and partition chromatography. It was found to be homogeneous by six additional criteria. In complement fixation experiments the synthetic product was immunologically active with antisera to HGH and [cys(Cam)53]-HGH-(134). Antiserum raised against the synthetic product was immunologically active in the homologous assay and with HGH,[Cys(Cam)53]-HGH-(1-134), and [Cys(Cam)53]-HGH-(15-125). The synthetic fragment exhibited 53% of the activity of [Cys(Cam)53]-HGH-(1-134) in the rat tibia assay.

Amino Acid Sequence↗

PubMed1978

Adhesion among neural cells of the chick embryo. III. Relationship of the surface molecule CAM to cell adhesion and the development of histotypic patterns.

We have previously identified a molecule (named cell adhesion molecule [CAM]) that is involved in the in vitro aggregation of neural cells from chick embryos. In the present report, specific anti-CAM antibodies have been used to demonstrated that CAM is localized in neural tissues, and is associated with the plasma membrane of retinal cells and neurites. Furthermore, it has been shown by antibody absorption techniques that the decreased adhesiveness of cultured retinal cells obtained originally from older embryos is correlated with a decrease in the density or accessibility of cell adhesion molecules on the surface of these cells. The central role of CAM in neural cell aggregation has been established by the observation that anti-CAM Fab' fragments inhibit adhesion between neural cells in a variety of assays. To investigate the function of CAM and cell adhesion in developing tissues, aggregates of retinal cells that are capable of forming histotypic patterns in vitro were cultured in the presence and absence of anti-CAM Fab'. The Fab' was found to inhibit sorting out of cell bodies and neurites and to decrease the number of membrane-membrane contacts, suggesting that CAM is associated with cell-cell, cell-neurite, and neurite-neurite interactions.

Animals↗

PubMed1976

The chick chorioallantoic membrane as a model system for the study of tissue invasion by viral transformed cells.

The chick chorioallantoic membrane (CAM) was used as an assay system to investigate the the invasive properties of viral transformed NIH/3Y3 cells. Scanning electron microscopy demonstrated that single Kirsten sarcoma virus (KiSV)-transformed cells passed between the epithelial cells of the CAM ectoderm within 6 hr of application, while viable NIH/3T3 cells did not penetrate the ectoderm within 24 hr. The transformed cells entered the mesoderm of the CAM and formed tumors of proliferating cells. The application of 5 X 10(5) KiSV-transformed cells resulted in the formation of donor cells resulted in the formation of the donor cell tumors within 5 days in 43% of the membranes. No tumors were formed when as many as 5 X 10(6) NIH/3T3 cells were applied to the membrane. NIH/3T3 cells transformed by the Abelson leukemia virus or the Moloney sarcoma virus also ivaded the CAM and formed tumors of proliferating cells within the mesoderm, while cells infected with the Moloney leukemia virus did not. NIH/3T3 cells inoculated onto the CAM 8 days after infection and transformation with KiSV formed tumors with a frequency similar to that of KiSV transformed cells that have been passaged in culture for many generations. Cells that formed invasive tumors within the mesoderm also attracted loops of host blood vessels.

Animals↗

PubMed1975

Transmission of marble spleen disease in turkeys and pheasants.

Marble spleen disease (MSD) of ring-necked pheasants (Phasianus colchicus L.) was transmitted in the laboratory to pheasants and turkeys (Meleagris gallopavo L.) by oral, colonic, and intravenous routes of inoculation, using cell-free supernatant fluids of splenic suspension from birds with naturally occurring MSD. Gross lesions consisted of large mottled spleens, a lesion more prominent in infected pheasants than in turkeys. The only microscopic lesion and also the criterion of infection were the presence of typical intranuclear inclusions of MSD. Similar to microscopic observations in naturally infected birds was the presence of inclusions in cells of spleen, bone marrow, liver, lung, bursa of Fabricius, and intestine-associated lymphoid tisse (IALT) OF EXPERIMENTALLY INFECTED BIRDS. Ultrastructural examination of splenic cells with intranuclear inclusions from turkey and pheasant with experimentally transmitted disease revealed viral particles and inclusions morphologically indistinguishable from those observed in naturally infected birds. Results of viral isolation procedures in turkey embryo chorioallantoic membrane (CAM) and yolk sac, as well as in turkey embryo fibroblast (TEF) and turkey kidney cell (TKC) cultures, were negative. Serologic assay by agar gel immunodiffusion tests indicated cross-reactivity between splenic MSD antigen and spleinic hemorrhagic enteritis (HE) of turkey antigen. Lines of fusion were formed between the splenic antigen from naturally occurring MSD in pheasants, experimentally transmitted MSD in turkeys, and HE of turkeys, using the homologous serum antibody to each of the 3 antigens.

Animals↗

PubMed1978

Growth hormone modulation of murine erythroleukemia cell growth in vitro.

There are few studies showing a biological effect of growth hormone (somatotropin) on cell proliferation in vitro at physiological concentrations. We report here that Friend virus-infected erythroleukemia cells are responsive to growth hormone in vitro. Using a serum-free clonogenic assay we found as little as 0.1 ng of human growth hormone per ml caused a prominent stimulation of cell proliferation. Peak activity of human growth hormone occurred at 200 ng/ml, resulting in a 2-fold increase in cloning. Human chorionic somatomammotropin and the Cys(Cam)53-hGH(1-134) fragment of human growth hormone were also active, but a biologically inert oxidized human growth hormone had no growth-promoting effect in vitro. Cell proliferation was stimulated by insulin with peak potentiation occurring at 1 ng/ml, and prolactin had a demonstrable stimulatory effect between 50 and 100 ng/ml. These observations indicate that growth hormone and related polypeptides have a direct effect on the in vitro proliferation of erythroleukemia cells in the absence of serum. The results confirm a direct action of growth hormone on mammalian cells and suggest that pituitary hormones may affect leukemic cell growth.

Animals↗

PubMed1978

Synthetic fragment of human growth hormone with hyperglycemic properties: residues 44-77.

Peptides homologous to Cys53 44-77 and Cys53 52-77 of the human growth hormone molecule were prepared by solid-phase synthesis and tested by our acute glucose tolerance test in ob/ob mice. Peptide 44-77, as either the Acm-Cys53 or Cam-Cys53 derivative, adversely affects glucose tolerance at doses of 100 to 150 nmol. Peptide 52-77 is nonhyperglycemic. Other diabetogenic properties are being tested.

Amino Acids↗

PubMed1979

Angiogenesis capacity as a diagnostic marker for human eye tumors.

Solid tumors have the capacity to continuously stimulate the proliferation of new capillaries. Aliquots (0.1 cc) of aqueous humor were aspirated from the anterior chamber of the eye of 38 patients undergoing elective ophthalmologic surgery. The material was lyophilized and then inplanted on the chorioallantoic membrane (CAM) of 10-day-old chick embryos for bioassay. The angiogenesis capacity of each sample was graded as negative or positive. Nine of 10 patients with histologically proven retinoblastoma had positive vascular responses. Seven of 11 patients with choroidal malignant melanoma had a positive response to their aqueous humor on the CAM. Aqueous samples from eyes with an iris and ciliary body malignant melanoma and a metastatic breast carcinoma to the iris had a positive angiogenic response. By contrast, only one of 15 patients undergoing operation for cataracts, glaucoma, or other nonmalignant ocular disease showed an angiogenesis response. The one patient who a positive assay later developed lymphocytic leukemia. These studies show that certain intraocular tumors display angiogenesis capacity before clinically evident neovascular changes of floating tumor cells are seen. Patients without tumors showed no angiogenesis response.

Angiogenesis Inducing Agents↗

PubMed1978

The chick embryo choriallantoic membrane as a bioassay for angiogenesis factors: reactions induced by carrier materials.

A variety of filter materials, sponges, and gels were placed on the chick chorioallantoic membrane (CAM), the reactions of it investigated and compared with those induced by natural egg materials (white eggshell membrane, coagulated albumen and yolk). Independently of the kind and nature of the naterials the CAM reacted nearly regularly underneath these diverse materials with a proliferation of ectodermal cells, fibroblasts, and blood vessels forming a highly capillarized granulation tissue. The area of the CAM surrounding the foreign materials frequently displayed an increase of small blood vessels macroscopically discernible and showing a radial arrangement (spoke-wheel-appearance according to FOLKMAN 1974). It is concluded that this type of vascular reaction cannot be considered as a characteristic feature for the action of a special tumour angiogenesis factor, since it can be induced by a variety of stimuli leading to an inflammatory reaction in the CAM. For the detection of special angiogenetic activities an objective quantification of the vessel reactions is necessary under consideration of the reactions induced by the mere presence of such foreign materials.

Allantois↗

PubMed2026

KLHL17 as a Prognostic Indicator and Therapeutic Target in Cervical Cancer: A Comprehensive Analysis.

INTRODUCTION: This study aims to clarify the role of kelch like family member 17 (KLHL17) in cervical cancer (CESC) is unclear. OBJECTIVE: To clarify this uncertainty, our research employed bioinformatics analysis coupled with experimental corroboration. METHODS: We utilized the Cancer Genome Atlas (TCGA) database to assess the expression of KLHL17 in various cancers, specifically CESC, and to explore its association with clinical characteristics, diagnostic utility, and prognostic significance in CESC. The current investigation delved into the potential regulatory pathways related to KLHL17, examining its connection with the infiltration of immune cells, the expression of immune checkpoint genes, the status of microsatellite instability (MSI), and the efficacy of diverse therapeutic agents in CESC. The research analyzed KLHL17 expression patterns using single-cell sequencing data from CESC samples and investigated the genetic variations of KLHL17 within this context. KLHL17 expression was validated using GSE145372. The presence and levels of KLHL17 in different cell lines were validated through quantitative real-time PCR (qRT-PCR) assays. RESULTS: KLHL17 exhibited irregular expression profiles across various cancer types, including CESC. Furthermore, increased KLHL17 levels in CESC patients were significantly associated with a lower progression-free survival (PFS) rate (hazard ratio: 1.62; 95% confidence interval: 1.01-2.60, p = 0.044). Moreover, KLHL17 expression emerged as a distinct prognostic indicator for CESC patients (p = 0.031). It has been associated with various biological pathways, such as cytokine-cytokine receptor interaction, primary immunodeficiency, cell adhesion molecules (CAMs), chemokine signaling pathway, steroid hormone biosynthesis, and others. The expression levels of KLHL17 were found to correlate with the presence of immune cells, the expression of immune checkpoint genes, and the status of MSI within CESC. Furthermore, KLHL17 expression exhibited a significant and inverse correlation with XMD15-27, rTRAIL, Paclitaxel, tp4ek, and tp4ek-k6. Furthermore, KLHL17 was found to be significantly positively regulated in CESC cell lines. DISCUSSION: The findings suggest that KLHL17 is involved in the progression of CESC and may serve as a potential prognostic marker and therapeutic target. KLHL17's association with immune cell infiltration and immune checkpoint genes indicates a role in immuneevasion. Future research should focus on validating these findings through independent datasets and experimental studies to elucidate the molecular mechanisms underlying KLHL17's role in CESC progression and immune regulation. CONCLUSION: KLHL17 is a promising prognostic marker and potential therapeutic target in CESC.

Humans↗

PubMed1975

Marek's disease virus (Kekava strain) replication in chickens, chick embryos and cell cultures.

In the course of 12 passages of Marek's disease virus (MDV) strain Kekava (MDV-Kekava) in chickens, the morbidity varied greatly (from 23 to 50 percent). MDV-Kekava produced plaques in cultures of chick embryo kidney and adult chicken kidney cells and chick embryo fibroblasts (CEF). The virus adaptation to the cultures was very slow. MDV-Kekava induced the formation of pocks on the chorioallantoic membranes (CAM) of chick embryos but the proportion of embryos with CAM lesions did not exceed 24 percent. Serial passaging of the virus in chick embryos beyond the 5th passage was unsuccessful. The results of virus isolation in chickens, cell cultures and chick embryos indicate the possibility of a long-term latent virus carrier state in chickens without development of tumours.

Animals↗

PubMed1978

Hemopoiesis on macrophage-coated cellulose acetate membranes (CAMS) in mice: an immunological study.

Macrophage-coated cellulose acetate membranes (CAMS), implanted into the peritoneal cavities of sublethally irradiated mice, support the growth of hemopoietic colonies. To investigate the nature of the precursor cells (CFU-ML) which form colonies on CAMS, we pre-treated marrow cells with rabbit anti-mouse brain serum (RAMBS), a known anti-pluripotent stem cell (CFU-S) serum, plus complement (C) and studied the number of colonies formed and the distribution of their sizes among the various histological types. Marrow cells pre-treated with RAMBS + C, even with the opportunity for interaction with macrophages in vivo, did not form fewer or smaller colonies than those formed from CRS + C treated cells, suggesting that most of the CFU-ML are antigenically distinct from CFU-S.

Animals↗

PubMed1976

Characterization of measles viruses in establishment of persistent infections in human lymphoid cell line.

Human lymphoid cells (NC-37) were infected with attenuated measles vaccine virus (Schwarz, AIK-C, and CAM-70 strains), subacute sclerosing panencephalitis virus (Mantooth and Halle strains), neurovirulent TYCSA strain, and wild type virus (Edmonston and Toyoshima strains) at an input multiplicity of 0-01. These strains were divided into two groups by their capacity to establish carrier states. CAM-70, Toyoshima, and Edmonston strains did not set up persistent infections in NC-37 cells, whereas AIK-C strain induced chronic cyclic infection and the Schwarz, TYCSA, Mantooth and Halle strains could set up persistent infections and furthermore two types of persistent infections were recognizable. Cells persistently infected with Schwarz strain contained nucleocapsid structures in both nucleus and cytoplasm, and produced infectious virus of 10(4) to 10(5) p.f.u./ml over 100 days after the inoculation of the virus but the cap-formation of measles antigens on the cell membrane was seldom observed. However, in cells persistently infected with TYCSA strain, nucleocapsid structures were rarely observed in the nucleus, but the cap-formation of measles antigens on the cell membrane was often observed. The titre of carried virus was always higher than the number of cells in the range of 10(6) to 10(7) p.f.u./ml. Mantooth strain was similar to Schwarz strain and Halle strain was similar to TYCSA strain in the properties of their carrier states. These carrier states were stable and the cells grew normally for over one year.

Antigens, Viral↗