Cells In Proliferative Phase Research Articles

Curcumin nanoemulsions inhibit oral squamous cell carcinoma cell proliferation by PI3K/Akt/mTOR suppression and miR-199a upregulation: A preliminary study.

Accumulating evidence indicates that curcumin (CUR) has anticancer properties in various cancers including oral squamous cell carcinoma (OSCC), but CUR is greatly restricted in clinical studies and applications due to its low bioavailability. Interestingly, the bioavailability of CUR was found to be significantly improved using loaded lipid nanoemulsions (NEs). To investigate the effect of CUR-NEs on cell proliferation of OSCC HSC-3 cells in vitro, and explore the potential mechanism of this effect in a preliminary study. CUR-NEs exhibited significantly cytotoxic effects on OSCC cells in a dose-dependent manner, compared with the control. The percentage of cells in proliferative phases (S + G2/M) was gradually decreased in a dose- or time-dependent manner caused by CUR-NEs. Moreover, CUR-NEs downregulated the protein expression of PI3K/Akt/mTOR and upregulated the expression of miR-199a that targeted PI3K in a dose- or time-dependent manner in OSCC cells. Importantly, CUR-NEs cloud effectively counteract the influence on cell proliferation of OSCC cells and the proliferative phases of cell cycle caused by miR-199a inhibitor a time-dependent manner. This in vitro preliminary study indicated that CUR-NEs may be an effective therapeutic agent for OSCC. Such effects could be related to inhibition of OSCC cell proliferation by PI3K/Akt/mTOR suppression and miR-199a upregulation.

The kinesin spindle protein inhibitor filanesib enhances the activity of pomalidomide and dexamethasone in multiple myeloma.

Kinesin spindle protein inhibition is known to be an effective therapeutic approach in several malignancies. Filanesib (ARRY-520), an inhibitor of this protein, has demonstrated activity in heavily pre-treated multiple myeloma patients. The aim of the work herein was to investigate the activity of filanesib in combination with pomalidomide plus dexamethasone backbone, and the mechanisms underlying the potential synergistic effect. The ability of filanesib to enhance the activity of pomalidomide plus dexamethasone was studied in several in vitro and in vivo models. Mechanisms of this synergistic combination were dissected by gene expression profiling, immunostaining, cell cycle and short interfering ribonucleic acid studies. Filanesib showed in vitro, ex vivo, and in vivo synergy with pomalidomide plus dexamethasone treatment. Importantly, the in vivo synergy observed in this combination was more evident in large, highly proliferative tumors, and was shown to be mediated by the impairment of mitosis transcriptional control, an increase in monopolar spindles, cell cycle arrest and the induction of apoptosis in cells in proliferative phases. In addition, the triple combination increased the activation of the proapoptotic protein BAX, which has previously been associated with sensitivity to filanesib, and could potentially be used as a predictive biomarker of response to this combination. Our results provide preclinical evidence for the potential benefit of the combination of filanesib with pomalidomide and dexamethasone, and supported the initiation of a recently activated trial being conducted by the Spanish Myeloma group which is investigating this combination in relapsed myeloma patients.

Calcium alginate particles for the combined delivery of platelet lysate and vancomycin hydrochloride in chronic skin ulcers

The aim of the present work was the development of a powder formulation for the combined delivery of platelet lysate and of a model antibiotic drug, vancomycin hydrochloride (VCM), in chronic skin ulcers. Calcium alginate particles were prepared by freeze-drying beads obtained by ionic gelation method. The experimental conditions adopted permitted the complete loading of VCM and of PDGF AB, the growth factor chosen as representative of those contained in PL. Such particles where able to absorb PBS (mimicking wound exudate), to form a gel and to modulate the release of VCM and of PDGF AB. They are characterized by enhancement properties of human fibroblast proliferation due to PL presence. In particular, PL, when loaded in alginate particles, was able not only to increase the number of viable cells, but also the number of cells in proliferative phase. Such properties were comparable to those of fresh PL indicating the capability of calcium alginate particles to load PL bioactive substances without altering their activity. The formulation developed is characterized by an easier and a less painful administration with respect to traditional gauzes and semisolid preparations and permits the loading in the same dosage form of active substances of different nature avoiding eventual incompatibility problems.

Progesterone Activates Multiple Innate Immune Pathways in Chlamydia trachomatis‐Infected Endocervical Cells

Susceptibility to Chlamydia trachomatis infection is increased by oral contraceptives and modulated by sex hormones. We therefore sought to determine the effects of female sex hormones on the innate immune response to C. trachomatis infection. ECC-1 endometrial cells, pre-treated with oestradiol or progesterone, were infected with C. trachomatis and the host transcriptome analysed by Illumina Sentrix HumanRef-8 microarray. Primary endocervical epithelial cells, prepared at either the proliferative or secretory phase of the menstrual cycle, were infected with C. trachomatis and cytokine gene expression determined by quantitative RT-PCR analysis. Chlamydia trachomatis yield from progesterone-primed ECC-1 cells was significantly reduced compared with oestradiol-treated cells. Genes upregulated in progesterone-treated and Chlamydia-infected cells only included multiple CC and CXC chemokines, IL-17C, IL-29, IL-32, TNF-α, DEFB4B, LCN2, S100A7-9, ITGAM, NOD2, JAK1, IL-6ST, type I and II interferon receptors, numerous interferon-stimulated genes and STAT6. CXCL10, CXCL11, CX3 CL1 and IL-17C, which were also upregulated in infected secretory-stage primary cells, and there was a trend towards higher levels of immune mediators in infected secretory-phase compared with proliferative-phase cells. Progesterone treatment primes multiple innate immune pathways in hormone-responsive epithelial cells that could potentially increase resistance to chlamydial infection.

Influenza A virus reproduction in humanendometrial cells

Study of replication of influenza A viruses (IAV) of A(H3N2) and A(H1N1)pdm subtypes in human endometrial cells was performed. It was shown that endometrial infection with IAV strongly depends on the multiplicity of infection and causes substantial changes in functional activities of these cells. Moreover endometrial cells of proliferative phase (day 8) were more sensitive to IAV infection than cells from secretory phase of menstrual cycle on day 19th.

Epitestosterone and Testosterone have Similar Nonclassical Actions on Membrane of Sertoli Cells in Whole Seminiferous Tubules

Epitestosterone is the 17α-epimer of testosterone. This steroid possesses antiandrogenic activities. The mechanism of action of epitestosterone has not been elucidated. The aim of this study was to investigate the nonclassical effect of epitestosterone on the membrane of Sertoli cells in proliferative phase (rats aged 15 days) and in nonproliferative phase (rats aged 21 and 35 days). The membrane potential of Sertoli cells was recorded using a standard single microelectrode technique. Epitestosterone (0.5, 1, and 2 μM) or testosterone (1 μM) was administered alone and after infusion with flutamide (1 μM), verapamil (100 μM), or U-73122 (2 μM). The testes of rats aged 12-15 days were preincubated with 45Ca2+ with or without flutamide (1 μM) and incubated with epitestosterone (1 μM) or testosterone (1 μM). Epitestosterone and testosterone produced a depolarization in the membrane potential and increased the membrane input resistance on Sertoli cells from rats of all 3 ages. The effect of epitestosterone did not change after perfusion with flutamide. Epitestosterone increased 45Ca2+ uptake within 5 min and this effect was not inhibited by flutamide. The absence of an effect by flutamide suggests that epitestosterone acts independently of the intracellular androgen receptor. The depolarizing effect was inhibited by verapamil, a voltage-dependent calcium channel blocker, and by U-73122, a phospholipase C inhibitor. These results indicate that epitestosterone acts on the membrane via a nonclassical signaling pathway; the effect was similar to the testosterone action on membrane of Sertoli cells in whole seminiferous tubules from rat testes.

Transforming effect of PDGFRA gene mutant on the cell function in gastrointestinal stromal tumor

To explore the effect of malignant transformation of the L839P, a new mutation site of the PDGFRA gene, on the pathogenesis of gastrointestinal stromal tumors. All recombinant plasmids were stably transfected into CHO cells by liposomes. Western blotting was used to detect the expression of PDGFRA protein. The cell growth curve was plotted by cell counting. Flow cytometry was used to detect the cell cycle and apoptosis of CHO cell, respectively. The stably transformed cells were inoculated subcutaneously into the back of nude mice and the mice were used to observe the tumorigenesis. Transient transfection of the mutant-type plasmids of PDGFRA gene and the wild-type plasmids of kit gene into the CHO cells was performed. Western blot was used to detect the expression of kit protein and its phosphorylated forms. PDGFRA protein expressed in the negative control, experimental group and positive control, except the empty vector. The growth curve showed that it was accelerated in the experimental group and positive control. The ratios of cells in proliferative phase were 28.4% (blank), 24.5% (negative control), 43.8% (experimental group) and 40.9% (positive control). Their apoptotic indexes were 1.8%, 1.9%, 1.5% and 1.6%, respectively. After three weeks, tumors were observed in the nude mice of experimental group and positive control, inoculated with the stably transformed cells. Moreover, the expression of phosphorylated protein of kit was enhanced after cotransfection of the mutant-type plasmids of PDGFRA and the wild-type plasmid of kit. The PDGFRA mutant L839P is a gain-of-function mutation and has obviously malignant transforming effect on normal cells, and may activate kit protein accelerating the tumorigenesis. Gastrointestinal stromal tumors;

Elastin metabolism in pelvic tissues: Is it modulated by reproductive hormones?

The purpose of this study was to investigate the effect of relaxin on extracellular matrix protein expression in pelvic fibroblasts that were cultured from women with stress urinary incontinence compared with asymptomatic control subjects. Periurethral vaginal wall fibroblasts from premenopausal women with stress urinary incontinence and continent women (in both the proliferative and secretory phase of the menstrual cycle) were stimulated with increasing concentrations of relaxin (0-500 ng/mL). The supernatant was sampled for matrix metalloproteinase-2 and -9 by zymography. Tissue inhibitors of metalloproteinase-1 and -2 and alpha-1 antitrypsin were evaluated with Western blot. Total elastase activity was measured by generation of free amino groups from succinylated elastin. Increasing concentrations of alpha-1 antitrypsin were added to cell lysate to evaluate total elastase activity inhibition. Proliferative-phase stress urinary incontinence fibroblasts demonstrated an increase in matrix metalloproteinase-2 and no change in matrix metalloproteinase-9 and tissue inhibitors of metalloproteinase-1 and -2 expressions with increasing relaxin concentrations. Cells from control subjects showed increased expression of matrix metalloproteinase-2 and -9, but no change in tissue inhibitors of metalloproteinases. Secretory-phase stress urinary incontinence fibroblasts showed no response in matrix metalloproteinase or tissue inhibitors of metalloproteinase expressions with relaxin stimulation. Secretory-phase control fibroblasts reacted by increasing matrix metalloproteinase-2 and -9 and tissue inhibitors of metalloproteinase-2. With respect to total elastase activity and alpha-1 antitrypsin expression, increasing doses of relaxin appear to increase elastolytic activity in stress urinary incontinence cells by decreasing the expression of alpha-1 antitrypsin in proliferative phase cells or increasing the total elastase activity in secretory phase cells. Fibroblast total elastase activity was inhibited by increasing concentrations of alpha-1 antitrypsin. Elastase activity appears to be increased in relaxin-stimulated stress urinary incontinence fibroblasts by either decreased inhibitor (alpha-1 antitrypsin) production or increased elastase activity.

Bone marrow response to large volume blood collection in the horse

Evaluation of erythropoietic regeneration in horses is difficult unless serial bone marrow aspirates are performed. To investigate the acute and chronic erythropoietic regenerative response of equine bone marrow following acute removal or loss of blood, sequential bone marrow aspirates over 4 weeks were taken from the sternum of five horses from which 20 ml kg-1of blood had been removed. We found that the total number of erythroid cells counted (expressed as a percentage of the total number of erythroid and myeloid cells counted) expanded initially by 13·7 per cent within 3 days after blood removal, the erythroid response peaking by 9 days with a further 13·5 per cent increase. This peak coincided with the lowest M:E ratio. Concomitantly, a shift from proliferative phase cells to maturing phase cells occurred, which appeared to persist beyond 31 days post collection. Thus, we found that the equine bone marrow mounted a regenerative erythropoietic response more slowly than previously determined and, also, regeneration of the erythroid compartment was incomplete 31 days after blood removal of this magnitude.

Comparison of the Effects of Semi-Occlusive Polyurethane Dressings and Hydrocolloid Dressings on Dermal Repair: 1. Cellular Changes

The effects on dermal repair of two wound dressings, one the semi-occlusive polyurethane sheet Opsite, the other the hydrocolloid Granuflex, were compared in full-thickness excised lesions on porcine skin during the period from 5 d to 6 months after injury. Quantitative studies were made of changes in the populations of polymorphonuclear leucocytes, macrophages, fibroblasts, and endothelial cells. The progress of repair in the wounds covered with the semi-occlusive dressing showed a decrease in the number of inflammatory cells (polymorphonuclear leukocytes and macrophages) from 5 to 60 d, whereas the number of proliferative phase cells (fibroblasts and endothelial cells) increased from 5 to 7 d. The total cellularity per unit area showed an increase between 5 and 7 d, that is, during the proliferative phase of repair, and then progressively decreased as the proliferative phase was succeeded by the remodeling phase. In contrast, the repair process in the hydrocolloid-dressed wounds was more complex. The number of inflammatory cells remained relatively high throughout and there were consistently fewer endothelial cells present throughout. Fibroblast number showed an initial fall from 5 to 14 d but then started to increase in number from 21 to 60 d. This chronic inflammatory reaction appeared to be in response to particulate matter that had been incorporated into the wound bed and hypodermis, and was still apparent 6 months after injury, when hydrocolloid particles were detectable microscopically in the hypodermis.

Immunohistochemical investigations of steroid receptors in normal and neoplastic squamous epithelium of the uterine cervix

The proliferation of squamous cells of the vagina and cervix uteri is induced by steroid hormones during the menstrual cycle. However, carcinoma of the cervix cannot be influenced by any hormone therapy. Forty-four different cervical specimens (different days in the menstrual cycle of healthy women and those with dysplastic lesions and carcinomas of the cervix) have been tested for estrogen (ER) and progesterone (PR) receptor protein content by means of immunohistochemistry. The ER content of the squamous epithelium depends upon the menstrual cycle: in the early proliferative phase cells of all layers are negative. In the midphase of proliferation the basal and parabasal layers become positive, and in the secretory phase positive cell nuclei can be found up to the superficial layers. A weak reaction to ER staining is found only in mild dysplastic lesions of the uterine cervix; severe dysplastic forms and invasive carcinomas were all negative. No positive PR was found in any squamous cell tissue. Stroma cells of the uterine cervix showed different straining intensity for ER and PR, regardless of the menstrual cycle. The loss of ER in the neoplastic cell could be an explanation for three clinical experiences: premenopausal patients have no tumor progression of the cervix uteri despite normal ovarian function; the duration of survival shows no relation to the receptor status of cervical carcinomas; and antihormonal treatment of cervical carcinomas produces no appreciable therapeutic success.

Immunohistochemical investigations of steroid receptors in normal and neoplastic squamous epithelium of the uterine cervix

The proliferation of squamous cells of the vagina and cervix uteri is induced by steroid hormones during the menstrual cycle. However, carcinoma of the cervix cannot be influenced by any hormone therapy. Forty-four different cervical specimens (different days in the menstrual cycle of healthy women and those with dysplastic lesions and carcinomas of the cervix) have been tested for estrogen (ER) and progesterone (PR) receptor protein content by means of immunohistochemistry. The ER content of the squamous epithelium depends upon the menstrual cycle: in the early proliferative phase cells of all layers are negative. In the midphase of proliferation the basal and parabasal layers become positive, and in the secretory phase positive cell nuclei can be found up to the superficial layers. A weak reaction to ER staining is found only in mild dysplastic lesions of the uterine cervix; severe dysplastic forms and invasive carcinomas were all negative. No positive PR was found in any squamous cell tissue. Stroma cells of the uterine cervix showed different straining intensity for ER and PR, regardless of the menstrual cycle. The loss of ER in the neoplastic cell could be an explanation for three clinical experiences: premenopausal patients have no tumor progression of the cervix uteri despite normal ovarian function; the duration of survival shows no relation to the receptor status of cervical carcinomas; and antihormonal treatment of cervical carcinomas produces no appreciable therapeutic success.

Comparison of the Effects of Moist and Dry Conditions on Dermal Repair

The effects on dermal repair of moist conditions, achieved by covering excised wounds with the adhesive polyurethane dressing OPSITE, and dry conditions, achieved by exposure to air through dry gauze dressings, were compared in full thickness excised wounds on porcine skin during the period from 1 to 21 d after injury. Quantitative studies were made of changes in the populations of neutrophils, macrophages, fibroblasts, and endothelial cells. The number of inflammatory phase cells (neutrophils and macrophages) decreased more rapidly under moist conditions than under dry conditions. There was also a more rapid increase in the number of proliferative phase cells (fibroblasts and endothelial cells) in the moist wounds; by 5 d after injury 66% of the cells of the granulation tissue of the moist wounds were of this type, compared with only 48% of the cells of equivalent areas of the dry wounds. By 21 d after injury the number of fibroblasts in the granulation tissue of the moist wounds had fallen below that in the dry wounds, suggesting that progress from the proliferative into the remodelling phase of repair was more rapid in the moist wounds. It was concluded that there was an acceleration of the inflammatory and proliferative phases of dermal repair in wounds healing under moist conditions when compared with those healing under dry conditions.

The action of IAA and kinetin on the mitotic cycle of proliferative and stationary phase excised root meristems

The influence of kinetin and IAA on the G1 (presynthetic), S (DNA synthetic) and G2 (postsynthetic) periods of the mitotic cycle of actively dividing (proliferative phase) and initially nondividing (stationary phase) root meristem cells was determined with the use of tritiated thymidine ( 3H-T ) and autoradiography. Proliferative phase meristems were cultured for 72 h in White's medium with 2 per cent sucrose and kinetin or IAA or both. Pulse labeling with 3 H-T was performed at 72 h. The stationary phase meristems were induced by a 24 h deprivation of sucrose. These meristems were almost devoid of dividing and DNA synthesizing cells. After establishment of the stationary phase the roots were transferred to medium containing sucrose, 3H-T, and kinetin or IAA or both. Results obtained with proliferative phase cells suggested that kinetin slightly increased G2 and certainly the (G1 durations. IAA appeared to act primarily on the duration of S. The combination of kinetin and IAA produced all three results. The effect on stationary phase cells, i.e. cells that are retained in either G1 or G2 until provided sucrose, indicated that kinetin and IAA impaired the initiation of DNA synthesis of most but not all of the G1 cells. The combination of kinetin and IAA reduced the initiation of DNA synthesis by G1 cells and further impaired the progress of cells through S; G2 cells also displayed sensitivity to the combination of kinetin and IAA in that they entered mitosis at a low rate. The concentrations of kinetin and IAA used in these experiments were similar to those used to induce cell proliferation in differentiated areas of the pea root; therefore, it is probable that controlling sites or moieties are activated or derepressed in cells of differentiated tissue and inactivated or repressed in meristematic cells by the direct or indirect action of the chemicals.