Stimulate Cell Growth Research Articles

Biological characterization of dehydrated amniotic membrane allograft: Mechanisms of action and implications for wound care.

There is a growing clinical demand in the wound care market to treat chronic wounds such as diabetic foot ulcers. Advanced cell and tissue-based products (CTPs) are often used to address challenging chronic wounds where healing has stalled. These products contain active biologics such as growth factors and cytokines as well as structural components that support and stimulate cell growth and assist in tissue regeneration. This study addresses the in vitro biologic effects of a clinically available dehydrated amniotic membrane allograft (DAMA). The broad mechanism of action results from DAMA's biologic composition that leads to stimulation of cell migration cell proliferation, and reduction of pro-inflammatory cytokines. Results show that DAMA possesses growth factors and cytokines such as EGF, FGF, PDGFs, VEGF, TGF-β, IL-8, and TIMPs 1 and 2. Furthermore, in vitro experiments demonstrate that DAMA stimulates cell proliferation, cell migration, secretion of collagen type I, and the reduction of pro-inflammatory cytokines IL-1β, IL-6, and TNF-α. This study findings are consistent with the clinical benefits previously published for DAMA and other CTPs in chronic wounds suggesting that the introduction of DAMA to non-healing, complex wounds helps to improve the wound milieu by providing essential structural components, cytokines, and growth factors to create an appropriate environment for wound healing.

The association ofEGFrs2237051 variant, serum EGF levels and generalized aggressive periodontitis: a preliminary study

BackgroundEpidermal growth factor (EGF) is a pro-inflammatory small peptide that stimulates cell growth, proliferation and differentiation through binding to its receptor. EGF rs2237051 and serum EGF levels have been demonstrated to be related with a variety of diseases, including several tumors and inflammatory diseases. Therefore, this study aims to investigate the association of the EGF rs2237051 variant and serum EGF levels in Chinese patients with generalized aggressive periodontitis (GAgP).Material and MethodsA case-control study was conducted among 216 patients with GAgP and 138 healthy controls. The clinical parameters of plaque index, probing depth, attachment loss and bleeding index were recorded. The EGF rs2237051 polymorphism was genotyped using time-of-flight mass spectrometry, and serum EGF levels were determined. Logistic and linear regression models were used to investigate the association between the genotypes of EGF rs2237051, serum EGF levels and GAgP risk.ResultsThe AA genotype of EGF rs2237051 showed higher risk for GAgP than the combined genotypes GG and AG (adjusted OR = 1.65, 95% CI [1.06–2.57]). Increased serum EGF levels were associated with GAgP (adjusted OR = 1.18, 95% CI [1.14–1.22]). Moreover, the serum EGF level for the AA genotype was significantly higher than that for the AG/GG genotypes in patients with GAgP (adjusted β = 4.70, 95% CI [2.09–7.31]).ConclusionWe demonstrated that EGF rs2237051 variant and the increased level of serum EGF were associated with the risk of GAgP, the serum EGF was up-regulated in patients with GAgP. It was indicated that serum EGF might be a biomarker of GAgP and EGF rs2237051 may be related to the genetic background of GAgP.

Cell Growth Stimulation, Cell Cycle Alternation, and Anti-Apoptosis Effects of Bovine Bone Collagen Hydrolysates Derived Peptides on MC3T3-E1 Cells Ex Vivo.

Bovine bone collagen hydrolysates promote bone formation through regulating bone growth. However, the peptide sequences within these isolates have not been characterized. In this study, twenty-nine peptides from bovine bone collagen hydrolysates were purified and identified using nano-HPLC-MS-MS and Peak Studio analysis. HHGDQGAPGAVGPAGPRGPAGPSGPAGKDGR (Deamidation) and GPAGANGDRGEAGPAGPAGPAGPR (Deamidation) enhanced cell viability, inhibited apoptosis, and significantly altered the cell cycle of MC3T3-E1 osteoblast cells. These peptides were selected to perform molecular docking analysis to examine the mechanism underlying these bioactivities. Molecular docking analysis showed that these two peptides formed hydrophobic interactions and hydrogen bonds with epidermal growth factor receptor (EGFR) to activate the EGFR-signaling pathway, which may explain their bioactivity. These findings indicate that these and other similar peptides might be candidates for the treatment of osteoporosis.

SUN-LB27 Interleukin-8 - a Possible Target for Melanoma Treatment? In-Vitro Studies Based on Human Melanoma Cell Models

Previous clinical studies showed that menstruating females were better protected in melanoma than post-menopausal women and men of any age. In addition, epidemiological studies showed an increased male mortality in melanoma. But these studies did not correlate with steroid status in females. Our in-vitro study showed female sex hormone progesterone significantly inhibited human melanoma cell growth. Further in-vitro study showed that progesterone action was mediated by a specific suppression of pro-inflammatory cytokine IL-8. Our research also showed that addition of IL-8 (1 ng/ml) to melanoma cells stimulated cell growth (117%) and suppression of IL-8 by curcumin (100 μM) pre-treatment suppressed human melanoma cell growth (26%) in-vitro. This observation prompted us to check the effect of male sex hormones androstenedione (AD) and testosterone (T) on melanoma cell growth. AD and T also suppressed cell growth and IL-8 secretion, but not as significantly as that of progesterone. However, addition of progesterone (10 μM) along with androgens showed an additive effect on the inhibition of melanoma cell growth and suppression of IL-8 secretion. As steroids (P, AD, T) targeted IL-8 for their action, it was decided to check whether vitamin-D3 also targeted IL-8 secretion and cell growth. Active form of vit-D3 (25 μM) also suppressed IL-8 secretion and cell growth. But, addition of progesterone (50 μM) along with D3 significantly suppressed cell growth and IL-8 secretion. This brought IL-8 into focus as a key molecule regulating melanoma cell growth. In order to check whether IL-8 was the molecule involved in regulating melanoma cell growth, IL-8 rescue experiment after curcumin (25 μM) pre-treatment was carried out. IL-8 (100 ng/ml) was able to rescue cell growth completely after pre-treatment with curcumin, suggesting IL-8 was the molecule involved in regulating melanoma cell growth. Literature also suggested important role for IL-8 in regulating melanoma cell growth. Conditional expression of IL-8 in nude mouse by Dr. Singh et al., indicated in-vivo role of IL-8 in melanoma growth and metastasis. Conclusion: Both, in-vitro and in-vivo studies suggested an important role for IL-8 in regulating melanoma growth and metastasis. So, IL-8 could be targeted to arrest melanoma growth and metastasis in-vivo. Hence, IL-8 could be a potential target for melanoma treatment.

Constructing the constitutively active ribosomal protein S6 kinase 2 from Arabidopsis thaliana (AtRPS6K2) and testing its activity in vitro.

Ribosomal protein S6 (RPS6) is the only phosphorylatable protein of the eukaryotic 40S ribosomal subunit. Ribosomes with phosphorylated RPS6 can selectively translate 5’TOP-(5’-terminal oligopyrimidine)-containing mRNAs that encode most proteins of the translation apparatus. The study of translational control of 5’TOP-mRNAs, which are preferentially translated when RPS6 is phosphorylated and cease to be translated when RPS6 is de-phosphorylated, is particularly important. In Arabidopsis thaliana, AtRPS6 is phosphorylated by kinase AtRPS6K2, which should in turn be phosphorylated by upper level kinases (AtPDK1 – at serine (S) 296, AtTOR – at threonine (T) 455 and S437) for full activation. We have cloned AtRPS6K2 cDNA gene and carried out in vitro mutagenesis replacing codons encoding S296, S437 and T455 by triplets of phosphomimetic glutamic acid (E). After the expression of both natural and mutated cDNAs in Escherichia coli cells, two recombinant proteins were isolated: native AtRPS6K2 and presumably constitutively active AtRPS6K2(S296E, S437E, T455E). The activity of these variants was tested in vitro. Both kinases could phosphorylate wheat (Triticum aestivum L.) TaRPS6 as part of 40S ribosomal subunits isolated from wheat embryos, though the non-mutated variant had less activity than phosphomimetic one. The ability of recombinant non-mutated kinase to phosphorylate TaRPS6 can be explained by its phosphorylation by bacterial kinases during the expression and isolation steps. The phosphomimetically mutated AtRPS6K2(S296E, S437E, T455E) can serve as a tool to investigate preferential translation of 5’TOP-mRNAs in wheat germ cell-free system, in which most of 40S ribosomal subunits have phosphorylated TaRPS6. Besides, such an approach has a biotechnological application in producing genetically modified plants with increased biomass and productivity through stimulation of cell growth and division.

Stimulation of cell growth and neurogenesis using protein-functionalized microfibrous scaffolds and fluid flow in bioreactors

Microfibrous viscose-rayon scaffolds functionalized with poly-L-lysine and laminin were used to culture pheochromocytoma (PC12) cells in recirculation bioreactors under bulk laminar flow conditions. To determine the effects of continuous forced convective flow on cell viability, growth, and neurogenesis, the cultures were exposed to fluid-induced shear stresses of magnitude 2.7 × 10–5 to 4.6 × 10–3 Pa for up to 12 days. The results were compared with those obtained using static scaffold and static monolayer (2D) control cultures. Cell growth rates were 1.5- to 1.8-fold greater in the bioreactor scaffold cultures relative to the controls without flow. After treatment of the cells with nerve growth factor, development and extension of neurite networks and expression of several neural differentiation markers including β3-tubulin, shootin1, and EphA2 were also substantially enhanced in the presence of fluid convection. In the control cultures without fluid flow, cell growth and differentiation were greater in the static scaffold cultures than in the static 2D monolayers. This work demonstrates the significant beneficial effects of features of the physical culture environment, such as hydrodynamic conditions and cell attachment surface geometry, in regulating culture performance. Flow devices capable of supporting long-term cell culture while delivering controlled levels of hydrodynamic shear are a valuable practical tool for promoting nerve tissue development and regeneration in vitro.

Comparative analysis of the chemical constituents from the tuberous root and stem of Pueraria candollei var. mirifica and evaluation of their estrogenic activity

Background: Pueraria candollei var.mirifica (PC) is a tuberous plant enriched with bioactive phytoestrogens. PC is currently high demanded in the global markets as dietary supplement products, which resulted in a shortage of tuberous root from the natural resource and the field cultivation. Materials and Methods: We compare phytoestrogens contents from the tuberous root and stem of PC and their estrogenic activities on MCF-7 cells proliferation. Results: The root and stem of PC accumulated different contents of phytoestrogens. The highest amount of total isoflavonoids and total chromenes was found in the root bark of PC. However, the stem bark (10 μg/mL crude extract) which contains a high level of chromenes and low amount of isoflavonoids exhibited higher growth stimulation of MCF-7 cells (127.25% ± 4.43% relative proliferative effect [RPE] as compared to estradiol [10−10 M]) than root bark at the same concentration with 116.32% ± 3.59% RPE. After the solubilization of the ethanol extract with 10% (v/v) aqueous ethanol, the soluble fraction of crude extract of stem stimulated proliferation of MCF-7 cells without cell growth suppression effect at high concentration (100 μg/mL) as observed in the insoluble fraction of the extract. Conclusion: We suggested that the soluble part of crude extract might be less toxic than the ethanolic crude extract and its non-polar components. Our study opens the possibility of using the stem of PC as a new alternative source of bioactive phytoestrogens for dietary supplement products.

Application of nanoparticles (ZnO, TiO2 and CuO), a new opportunity for the stimulation of cell growth and azadirachtin production in cell suspension culture of Azadirachta indica

Nanoparticles have unique physicochemical properties and provide great opportunities in plant science studies. In this study, we investigated the impact of ZnO, TiO2 and CuO nanoparticles (0, 20, 40, 60, and 80 mg/L) and sampling times (2, 4, and 6 days) on cell suspension growth and azadirachtin accumulation and production. Factorial experiments based on a completely randomized design with three replications were used. Results demonstrated that different nanoparticles had a different effect on the studied characters. When ZnO nanoparticles were used, the highest fresh (540.73 g/L), dry cell weight (15.93 g/L), azadirachtin accumulation (5.15 mg/g DW) and production (68.27 mg/L) were obtained at control condition, 80 and 40 mg/L ZnO nanoparticles, and control condition after 6 days, respectively. The highest amount of fresh (526.95 g/L) and dry (17.05 g/L) cell weight and azadirachtin production (82.21 mg/L) and accumulation (5.93 mg/g DW) were observed in 20 mg/L TiO2 nanoparticles, 40 mg/L of TiO2 nanoparticles after 2 days, 20 mg/L TiO2 nanoparticles in 4 days and 60 mg/L of TiO2 nanoparticles, respectively. With applying CuO nanoparticles, the highest fresh cell weight and azadirachtin accumulation were 422.59 g/L and 4.00 mg/L, achieved in control conditions respectively. Also, the highest amount of azadirachtin production was 68.27 mg/L, observed in control conditions on the 6th day of treatment. It seems that suitable cell growth, except in some cases, occurred in the absence of elicitors, but azadirachtin accumulation and production were stimulated by nanoparticles treatment. However, the results showed that the CuO nanoparticles caused a decrease in overall azadirachtin accumulation and production in the cells.

Can Interleukin‐8 Alone be Responsible for Regulating Melanoma Growth & Metastasis? In‐vitro studies based on treatments of human melanoma cell lines (BLM and 1205Lu) with sex steroids, IL‐8, curcumin and active vitamin‐D3

Previous clinical studies indicated that menstruating females were better protected in melanoma than post‐menopausal women and men of any age. But, the mechanism responsible for protection in menstruating females was not investigated. In addition, epidemiological data showed an increased male mortality in melanoma. Our in‐vitro study showed female sex hormone progesterone significantly inhibited human melanoma cell growth. Elisarray showed that progesterone action was mediated by a specific suppression of pro‐inflammatory cytokine IL‐8. So, the effect of male sex hormones androstenedione (AD) and testosterone (T) on melanoma cell growth was checked. AD and T also suppressed cell growth and IL‐8 secretion. But, addition of progesterone (10μM) along with androgens showed an additive effect on the inhibition of melanoma cell growth and IL‐8 suppression. This observation gave the idea of protective function of progesterone in melanoma, which was experimentally observed by a significant decrease in in‐vitro migration and adhesion functions. As steroids (P, AD, T) targeted IL‐8 for their actions, effect of vit‐D3 on melanoma cells was checked. Active form of vitamin‐D3 (25 μM) also targeted IL‐8 and decreased cell growth, suggesting IL‐8 could be regulating melanoma cell growth. In order to check whether IL‐8 regulated melanoma cell growth, IL‐8 was directly added to melanoma cells and endogenous IL‐8 was suppressed by 2 hrs pre‐treatment with curcumin. Addition of IL‐8 (1 ng/ml) to 1205Lu melanoma cells stimulated cell growth to 117%, whereas suppression of IL‐8 by curcumin (100 μM) pre‐treatment decreased cell growth to 24%. Similar results were obtained with BLM cells. This brought IL‐8 into focus as an important molecule regulating melanoma cell growth. Further in‐vitro experiments to restore curcumin (25 μM) pre‐treated melanoma cell growth (71.5%) by the addition of IL‐8 (100 ng/ml) resulted in a near complete restoration of cell growth (95.3%) in 1205Lu melanoma cells, suggesting IL‐8 could be the molecule regulating melanoma cell growth. Similar results were observed with BLM cells. In addition, in‐vivo conditional expression of IL‐8 in nude mouse by Dr. Singh et al., indicated involvement of IL‐8 in melanoma growth and metastasis.ConclusionBoth in‐vitro and in‐vivo studies indicated that IL‐8 alone could be responsible for regulating melanoma growth and metastasis. Hence, IL‐8 could be targeted to arrest melanoma growth and metastasis in‐vivo.

Insect cytokine growth-blocking peptide may regulate density-dependent phase trait of cuticular melanization in the larval armyworm, Mythimna separata

Abstract Growth-blocking peptide (GBP) is a 25 amino acid insect cytokine found in lepidopteran insects that has diverse biological activities such as larval growth regulation, paralysis induction, cell proliferation and stimulation of immune cells. Density-dependent phase polyphenism is a phenomenon of phenotypic plasticity in which the expression of a variety of traits can be affected by local population density. In the present study, the armyworm Mythimna separata larvae with four rearing densities (1 larva/vial, 2 larvae/vial, 4 larvae/vial and 6 larvae/vial) were tested for cuticular melanization and body weight throughout the third-fifth instar, and the functional role of GBP in regulating the changes was investigated. The results indicated that when reared at high densities, the larvae exhibited less body weight and more degree of cuticular melanization than larvae reared at low densities. The gene expression of GBP in armyworm larvae showed an initial rise and then decline trend with increased rearing densities in the third to fifth instar. Compared with control, more degree of cuticular melanization was observed in GBP-injected larvae (500 ng/larva in volume 50 μL) than that in Ringer’s solution-injected counterparts. Furthermore, the gene expression level of dopa decarboxylase and prophenoloxidase increased significantly in GBP-injected fifth instar larvae from 6 h to 12 h after injection, suggesting the role of GBP in modulating density-dependent phase trait of armyworm cuticular melanization.

Molecular Engineering Strategies Tailoring the Apoptotic Response to a MET Therapeutic Antibody.

The MET oncogene encodes a tyrosine kinase receptor involved in the control of a complex network of biological responses that include protection from apoptosis and stimulation of cell growth during embryogenesis, tissue regeneration, and cancer progression. We previously developed an antagonist antibody (DN30) inducing the physical removal of the receptor from the cell surface and resulting in suppression of the biological responses to MET. In its bivalent form, the antibody displayed a residual agonist activity, due to dimerization of the lingering receptors, and partial activation of the downstream signaling cascade. The balance between the two opposing activities is variable in different biological systems and is hardly predictable. In this study, we generated and characterized two single-chain antibody fragments derived from DN30, sharing the same variable regions but including linkers different in length and composition. The two engineered molecules bind MET with high affinity but induce different biological responses. One behaves as a MET-antagonist, promoting programmed cell death in MET “addicted” cancer cells. The other acts as a hepatocyte growth factor (HGF)-mimetic, protecting normal cells from doxorubicin-induced apoptosis. Thus, by engineering the same receptor antibody, it is possible to generate molecules enhancing or inhibiting apoptosis either to kill cancer cells or to protect healthy tissues from the injuries of chemotherapy.

Investigation of the role of β-TrCP in growth hormone transduction defect (GHTD).

Background Growth hormone(GH) and epidermal growth factor (EGF) stimulate cell growth and differentiation, and crosstalking between their signaling pathways is important for normal cellular development. Growth hormone transduction defect (GHTD) is characterized by excessive GH receptor (GHR) degradation, due to over-expression of the E3 ubiquitin ligase, cytokine inducible SH2-containing protein (CIS). GH induction of GHTD fibroblasts after silencing of messenger RNA (mRNA) CIS (siCIS) or with higher doses of GH restores normal GH signaling. β-Transducing-repeat-containing protein (β-TrCP), another E3 ubiquitin ligase, also plays a role in GHR endocytosis. We studied the role of β-TrCP in the regulation of the GH/GHR and EGF/EGF receptor (EGFR) pathways in normal and GHTD fibroblasts. Materials and methods Fibroblast cultures were developed from gingival biopsies of a GHTD (P) and a control child (C). Protein expression and cellular localization of β-TrCP were studied by Western immunoblotting and immunofluorescence, respectively, after: (1) GH 200 μg/L human GH (hGH) induction, either with or without silence CIS (siCIS), and (2) inductions with 200 μg/L GH or 1000 μg/L GH or 50 ng/mL EGF. Results After induction with: (1) GH200/siCIS, the protein expression and cytoplasmic-membrane localization of β-TrCP were increased in the patient, (2) GH200 in the control and GH1000 in the patient, the protein and cytoplasmic-membrane localization of β-TrCP were increased and (3) EGF, the protein expression and cytoplasmic-membrane localization of β-TrCP were increased in both the control and the patient. Conclusions (1) β-TrCP appears to be part of the negative regulatory mechanism of the GH/GHR and EGF/EGFR pathways. (2) There appears to be a negative correlation between β-TrCP and CIS. (3) In the control and GHTD patient, β-TrCP increases when CIS is suppressed, possibly as a compensatory inhibitor of the GH/GHR pathway.

Iron and hepcidin mediate human colorectal cancer cell growth

High dietary iron intake is a risk factor for the development of colorectal cancer. However, how iron subsequently impacts the proliferation of colorectal cancer cells remains unclear. This study determined the expression of six iron regulatory genes in twenty-one human colorectal cancer (CRC) biopsies and matched normal colonic tissue. The results show that only hepcidin and ferritin heavy chain expression were increased in CRC biopsies as compared to matched normal tissues. Four established human CRC cell lines, HT-29, HCT-116, SW-620 and SW-480 were subsequently examined for their growth in response to increasing concentrations of iron, and iron depletion. Real time cell growth assay showed a significant inhibitory effect of acute iron loading in HCT-116 cells (IC50 = 258.25 μM at 72 h), and no significant effects in other cell types. However, ten week treatment with iron significantly reduced HT-29 and SW-620 cell growth, whereas no effect was seen in HCT-116 and SW-480 cells. Intracellular labile iron depletion induced the complete growth arrest and detachment of all of the CRC cell types except for the SW-620 cell line which was not affected in its growth. Treatment of starved CRC cells with hepcidin, the major regulator of iron metabolism, induced a significant stimulation of HT-29 cell growth but did not affect the growth of the other cell types. Collectively these results show that iron is central to CRC cell growth in a manner that is not identical between acute and chronic loading, and that is specific to the CRC cell type.

Nanotechnology Scaffolds for Alveolar Bone Regeneration.

In oral biology, tissue engineering aims at regenerating functional tissues through a series of key events that occur during alveolar/periodontal tissue formation and growth, by means of scaffolds that deliver signaling molecules and cells. Due to their excellent physicochemical properties and biomimetic features, nanomaterials are attractive alternatives offering many advantages for stimulating cell growth and promoting tissue regeneration through tissue engineering. The main aim of this article was to review the currently available literature to provide an overview of the different nano-scale scaffolds as key factors of tissue engineering for alveolar bone regeneration procedures. In this narrative review, PubMed, Medline, Scopus and Cochrane electronic databases were searched using key words like “tissue engineering”, “regenerative medicine”, “alveolar bone defects”, “alveolar bone regeneration”, “nanomaterials”, “scaffolds”, “nanospheres” and “nanofibrous scaffolds”. No limitation regarding language, publication date and study design was set. Hand-searching of the reference list of identified articles was also undertaken. The aim of this article was to give a brief introduction to review the role of different nanoscaffolds for bone regeneration and the main focus was set to underline their role for alveolar bone regeneration procedures.

Role of insulin-like growth factor-1 receptor in female androgenetic alopecia

Objectives To find if there is a role of insulin-like growth factor-1 receptor (IGF-1R) in female androgenetic alopecia (FAGA). Background Patterned hair loss continues to be one of the most important hair problems affecting both men and women. It is the most common form of alopecia, affecting up to 50% of women in the course of their lives. Insulin-like growth factor-1 (IGF-1) is one of the most potent natural activators of the protein kinase B signaling pathway, a stimulator of cell growth and proliferation, and a potent inhibitor of programed cell death. IGF-1 is a physiological regulator of hair growth cycle. It helps maintain the anagen stage. Patients and methods This case–control study was conducted on 20 FAGA patients and 10 age-matched healthy female patients with no androgenetic alopecia as control. They were chosen from Dermatology Clinic at Menoufia University Hospital between April 2017 to April 2018.They were subjected to history taking and complete medical examination. Punch biopsies were taken, and by immunohistochemical staining, the degree of intensity and the percentage of IGF-1R were determined. Results The degree of intensity and percentage of IGF-1R in the diseased patients showed statistically significant difference from the normal patients (P = 0.001). There was no significant correlation between IGF-1R expression and age (P = 0.120) and menstrual history (P = 0.189), respectively. Conclusion On the basis of the results, it could be concluded that IGF-1R markedly diminished in FAGA.

Role of oncoviruses in oncogenesis

The author describes DNA oncoviruses and RNA oncoviruses, their ways of infiltrating the host’s cells, and the possibilities of neoplastic transformation of cells by these microorganisms. The role of protooncogenesis and oncogenesis in both humans and animals is discussed. The transformation of cells by viruses is normally insufficient for oncogenesis; the cells also need to gain “immortality,” which usually requires 4-5 genetic changes (the so-called clonal development of cells), (Fig. 1). Oncoviruses remove suppressor growth factors while enhancing the effects that stimulate cell growth through e.g. hormones, cytokines, or transcription activators. In addition, the author discusses the role of the optimization principle in neogenesis.

Difficult to Heal Wounds Intralesional Insulin Therapy Trial

Introduction-Insulin exerts metabolic and growth-stimulating actions both through its own receptors and the receptors of its homologous factors (IGFs), although with different degrees of affinity. The A receptor of insulin acts more intensely on the cell membrane, with a metabolic response, whereas the B receptor is quickly internalized, stimulates cell growth, can be directed to the treatment of wounds difficult to heal. Objective-we proposes to evaluate the potential of insulin as an inducing agent in the regeneration of wounds that are difficult to heal as a therapeutic option. Methodology-Simultaneous intra and perilesional injections of 0.01ml of the solution containing 5UI of NPH single-component insulin diluted in 1 ml of lidocaine into various planes and wound sites. Clinical, glycemic and photographic evaluations of the patients were performed. Results-The cicatricial response occured in all patients and observed since the first applications. Withouth glycemic changes. Comments - The responses obtained can be attributed to both the stimulation of insulin receptors and that of their counterparts. Conclusions-The method was effective and without adverse effects. Different intervals between applications did not change the results. The availability of insulin and safety for its use indicate the method as a therapeutic option in difficult to heal wounds.

Cell Wall Membrane Fraction of Chlorella sorokiniana Enhances Host Antitumor Immunity and Inhibits Colon Carcinoma Growth in Mice.

A colon cancer growth inhibitor partially purified from the isolated cell wall membrane fraction of Chlorella sorokiniana, here referred to as Chlorella membrane factor (CMF), was evaluated for its antitumor and immunomodulatory effects in cell culture and in a colon carcinoma mouse model. The CMF treatment dose- and time-dependently inhibited colon carcinoma cell growth in 2-dimensional cultures. Treatment with CMF also significantly inhibited the growth of colon carcinoma spheroids in 3-dimensional cell culture in coculture with T lymphocytes. In a mouse CT26 colon carcinoma peritoneal dissemination model, intraperitoneal injection of CMF (10 or 30 mg dry weight/kg body weight, every other day) dose-dependently and significantly attenuated the growth of tumor nodules via induction of tumor cell apoptosis. Evaluation of immune cell populations in ascites showed that CMF treatment tended to increase T lymphocytes but lower granulocyte populations. The present study suggests that the cell wall membrane fraction of Chlorella sorokiniana contains a bioactive material that inhibits colon carcinoma growth via direct cell growth inhibition and stimulation of host antitumor immunity. Hence, it is suggested that the Chlorella cell wall membrane extract or a bioactive substance in the extract is an attractive complementary medicine for cancer therapy.

In Silico Study Predicts CCDC69 as a Novel Tumour Suppressor Gene in HER2+ Breast Cancer

Differential gene expression analysis using databases followed by overall survival (OS) analysis is currently used to identify different oncogenes and tumour suppressor genes. The present study identified coiled coiled domain containing protein 69 (CCDC69) as a tumour suppressor gene in breast cancer by differential gene expression analysis using TCGA dataset for breast adenocarcinoma (BRCA) followed by OS and relapse free survival (RFS) analysis using Kaplan Meier (KM) plotter tool. CCDC69 was observed to be down regulated in tumour of breast cancer patients in BRCA. Following OS analysis for different breast cancer sub-types, low expression of CCDC69 has been observed to be associated with poor survival in HER2+ breast cancer only. CCDC69 was also found to be down regulated in different HER2+ breast cancer cells by analysing Gene Expression Omnibus (GEO) database. Additionally, CCDC69 was found to be under expressed in single cell HER2 positive population, which is evident from the single cell expression ATLAS database. Furthermore, CCDC69 has been observed to be lowly expressed with overexpression of HER2 in breast cancer by co-expression study. The possible mechanism of CCDC69 down regulation in HER2+ breast cancer was resolved using P-SCAN tool. P-SCAN analysis suggested a group of transcription factors (TFs) among which androgen receptor (AR) has been selected as the probable TF that could play a role in CCDC69 down regulation in HER2+ breast cancer. Moreover, overexpression of AR has been observed in BRCA and HER2+ single cell population. AR has also been observed to be co-expressed positively with HER2, but negatively with CCDC69 in breast cancer. Down regulation of CCDC69 can be predicted to stabilize microtubule formation following stimulation of cell growth and cell migration leading to HER2+ breast cancer progression and metastasis.

Stimulation of cell growth by addition of tungsten in batch culture of a methanotrophic bacterium, Methylomicrobium alcaliphilum 20Z on methane and methanol

It is carried out for researches to convert methane, the second most potent greenhouse gas, to high-value chemicals and fuels by using methanotrophs. In this study, we observed that cell growth of Methylomicrobium alcaliphilum 20Z in the batch cultures on methane or methanol was stimulated by the addition of tungsten (W) without formate accumulation. Not only biomass yield but also the total products yield (biomass and formate) on carbon basis increased up to 11.50-fold and 1.28-fold respectively in W-added medium. Furthermore, a significant decrease in CO2 yield from formate was observed in the W-added cells, which indicates that W might have affected the activity of certain enzymes involved in carbon assimilation as well as formate dehydrogenase (FDH). The results of this study suggest that M. alcaliphilum 20Z is a promising model system for studying the physiology of the aerobic methanotroph and for enabling its industrial use for methane conversion through high cell density cultivation.