Natural Herbal Estrogen-Mimetics (Phytoestrogens) Promote the Differentiation of Fallopian Tube Epithelium into Multi-Ciliated Cells via Estrogen Receptor Beta.

Phytoestrogens are herbal polyphenolic compounds that exert various estrogen-like effects in animals and can be taken in easily from a foodstuff in daily life. The fallopian tube lumen, where transportation of the oocyte occurs, is lined with secretory cells and multi-ciliated epithelial cells. Recently, we showed that estrogen induces multi-ciliogenesis in the porcine fallopian tube epithelial cells (FTECs) through the activation of the estrogen receptor beta (ERß) pathway and simultaneous inhibition of the Notch pathway. Thus, ingested phytoestrogens may induce FTEC ciliogenesis and thereby affect the fecundity. To address this issue, we added isoflavones (genistein, daidzein, or glycitin) and coumestan (coumestrol) to primary culture FTECs under air-liquid interface conditions and assessed the effects of each compound. All phytoestrogens except glycitin induced multi-ciliated cell differentiation, which followed Notch signal downregulation. On the contrary, the differentiation of secretory cells decreased slightly. Furthermore, genistein and daidzein had a slight effect on the proportion of proliferating cells exhibited by Ki67 expression. Ciliated-cell differentiation is inhibited by the ERß antagonist, PHTPP. Thus, this study suggests that phytoestrogens can improve the fallopian tube epithelial sheet homeostasis by facilitating the genesis of multi-ciliated cells and this effect depends on the ERß-mediated pathway.

Curcumin nanoparticles incorporated in PVA/collagen composite films promote wound healing.

Skin repair remains a common problem in plastic surgery. Wound dressing plays an important role in promoting local skin healing and has been widely studied. This study aimed to manufacture a composite film (CPCF) containing curcumin nanoparticles, collagen, and polyvinyl alcohol (PVA) to effectively promote the healing of skin wounds. Sustained drug release from the composite film provides long-term protection and treatment for skin wounds. Both antibacterial property and good histocompatibility of the CPCF were examined by analyzing antibacterial activity and cytotoxicity to validate its applicability for wound management. Moreover, in vivo studies proved that the CPCF had a rapid healing rate of 98.03%±0.79% and mature epithelialization on day 15 after surgery. Obvious hair follicles and earlier re-epithelialization was also noticed in the CPCF group using H&E staining. The result of Masson's trichrome staining confirmed that CPCF could promote the formation of collagen fibers. In summary, CPCF may be promising as a wound dressing agent in wound management owing to its rapid wound-healing effects.

Effects of iron on intestinal development and epithelial maturation of suckling piglets.

The purpose of the present study was to discover the effects of iron on the intestinal development and epithelial maturation of suckling piglets. Twenty-seven newborn male piglets from 9 sows (3 piglets per sow), with similar body weight, were selected. The 3 piglets from the same sow were randomly divided into 1 of the 3 groups. The piglets were orally administrated with 2 mL of normal saline (CON group) or with 25 mg of iron by ferrous sulfate (OAFe group; dissolved in normal saline) on the 2nd, 7th, 12th, and 17th day, respectively, or intramuscularly injected with 100 mg of iron by iron dextran (IMFe group) on the 2nd day. The slaughter was performed on the 21st day and intestinal samples were collected. Compared with the CON group, iron supplementation significantly increased the length (P < 0.001), weight (P < 0.001), relative weight (P < 0.001), and the length:weight ratio (P < 0.001) of the small intestine in both OAFe and IMFe groups. The villus height (P < 0.001), crypt depth (CD) (P < 0.001), villus width (P = 0.002), and surface area (P < 0.001) in the jejunum of IMFe and OAFe piglets were also greater than those in CON piglets. The mRNA expression of trehalase (Treh; P = 0.002) and sucrase isomaltase (Sis; P = 0.043), markers of epithelial maturation, increased in OAFe and IMFe piglets, respectively. Moreover, enterocyte vacuolization, observed in fetal-type enterocyte, was reduced in OAFe and IMFe piglets, compared with CON piglets. However, no significant difference in the expression of the target genes of wnt/ß-catenin signaling pathway was observed. The results indicated that both oral administration and intramuscular injection with iron promoted intestinal development and epithelial maturation in suckling piglets and that the effects of iron may be independent of wnt/ß-catenin signaling.

Beneficial effect of Curcumin Nanoparticles-Hydrogel on excisional skin wound healing in type-I diabetic rat: Histological and immunohistochemical studies.

Management of diabetic wounds remains a major challenge in the medical field, mostly due to incompetent outcomes of treatments. Curcumin has been documented as anti-inflammatory, antioxidant, antimicrobial and antineoplastic agent in addition to wound healing activities. However, its poor aqueous solubility and impaired skin permeation handicap its topical pharmaceutical usage. Hydrogel loaded curcumin nanoparticle (Cur-NP/HG) could overcome this pitfall and enable extended topical delivery of curcumin. Rat model of diabetes mellitus (DM) type I was induced using single injection of 70mg/kg streptozotocin (STZ) followed by full thickness skin wound. Rats were divided into 4 groups. GpI: control non-diabetic, GpII: diabetic non-treated, GpIII: diabetic treated with topical curcumin hydrogel (Cur/HG) and GpIV: diabetic treated with topical Cur-NP/HG. Histological assessment of epidermal regeneration, dermo-epidermal junction, leukocyte infiltration and collagen deposition, in addition to immunohistochemical staining for vascular endothelial growth factor (VEGF) and aquaporin-3 (AQP3) were performed. Diabetic rat possessed impaired wound closure, persistence of inflammation and decreased collagen deposition as compared to non-diabetic control. Application of Cur/HG induced partial improvement of the healing process in diabetic rats. Cur-NP/HG treatment provoked obvious improvement of the healing process with complete re-epithelization, intact dermo-epidermal junction, reorganization of the dermis with significantly increased collagen deposition and VEGF and AQP3 expression. These results illustrated that Cur-NP/HG have effectively improved the healing process in diabetic skin wound with substantial differences in the wound healing kinetics compared to wounds that received Cur/HG.

Review: Exogenous butyrate: implications for the functional development of ruminal epithelium and calf performance.

The importance of the use of exogenous butyrate in calves' diets is due to its role as a factor stimulating the functional development of ruminal epithelium and improving calf performance during the transition from preruminant to ruminant status. Our review will first present results related to effects of the administration of butyrate in calves' diets on the development of ruminal epithelium toward a more effective absorption and metabolism of fermentation products from the rumen. The introduction of sodium butyrate at a level of about 0.3% of diet dry matter is accompanied by an increase to 35% in butyrate concentration in the rumen of 33-day-old calves. Mutual reliance between an enhanced ruminal concentration of butyrate and the activities of transcription factors, genes and proteins involved in cell proliferation, ketogenesis and the maintenance of cell pH homeostasis in the ruminal epithelial cells has been clearly confirmed in many experiments. Second, the review presents results related to the effects of the introduction of butyrate salts in the diet on calf performance. Of 11 studies a positive effect was found in six; five of these were obtained from the calves that started receiving butyrate supplement at a level of about 0.3% diet dry matter from the age of 3 to 5 days. Results indicate that when a supplement is given to calves soon after birth the functional development of ruminal epithelium in cooperation with the endocrine and digestion systems is transferred into improving the efficiency of rearing. There have been no studies on the effects of greater amounts of butyrate salts in milk replacer; butyrate constitutes about 1.2% of the whole cow's milk dry matter. In older calves, when butyrate administration is provided as a component of a starter concentrate at the increasing inclusion rate from 0.3% to 3.0%, the practical effect in calf performance relates to the risk of depression of rumen pH below 5.5 and accompanying disruption of the organization of the ruminal epithelial tissue. The higher risk is noted in calves received starter with substantial content of a rapidly degradable starch. At present, the insufficient number of positive results confirming the beneficial effect of butyrate supplements in terms of an improvement in performance does not allow their recommendation for use in the practical feeding of calves.

Antioxidant activity in plasma and rumen papillae development in lambs fed fermented apple pomace.

The effect of fermented apple pomace (FAP) on animal health, antioxidant activity (AA), hematic biometry (HBm) and the development of ruminal epithelium were investigated in a study with 24 finishing lambs (BW = 25.4 ± 3.3 Kg). Lambs were grouped by sex (12 male and 12 female) and fed (n = 6 per group of treatment) a basal fattening diet (Control diet, T1) or the basal diet supplemented to include 10.91% of fermented apple pomace (FAP diet, T2). The animals were kept 56 d in individual metabolic cages, with ad libitum access to water and feed. Two blood samples were collected from each animal on d 0, 28, and 56 to determine AA in plasma and hematic biometry (HBm). Four samples of ruminal tissue were taken postmortem to evaluate the development of ruminal epithelium based on the length (LP) and width (WP) of papillae. AA and HBm data were analyzed with a mixed model (fixed effects: diet, sampling, sex, and their interaction; using the experimental unit nested in the effect of the diet as the random effect). LP and WP were analyzed with a hierarchical model, as simple and nested effects in the sampling site, where the fixed effects were the diet and the sex of the animal and their interaction. There was an effect of diet on AA, which was higher (P < 0.06) in T2 vs. T1 at 56 d (24.34 vs. 21.79 mM Fe2). Leukocytes increased (P < 0.05) from 7.52*10(3) ± 1.29*10/(3)µL to 9.14*10(3) ± 1.24*10(3)/µL in all the animals in the experiment, with a marked increased (P < 0.05) at 28 d after beginning of the feeding period, with values within the normal range for this species and without effect of the diet (P > 0.05) for the other indicators of HBm. Males' LP was higher in T2 than in T1 (P < 0.05). It was concluded that the use of FAP in the diets of finishing sheep reaped benefits on animal health and the development of rumen epithelium by improving antioxidant activity in plasma and stimulating the growth of papillae.

The chiton stylus canal: An element delivery pathway for tooth cusp biomineralization.

A detailed investigation of the stylus canal situated within the iron mineralized major lateral teeth of the chiton Acanthopleura hirtosa was undertaken in conjunction with a row-by-row examination of cusp mineralization. The canal is shown to contain columnar epithelial tissue similar to that surrounding the mineralized cusps, including the presence of iron rich particles characteristic of the iron storage protein ferritin. Within the tooth core, a previously undescribed internal pathway or plume is evident above the stylus canal, between the junction zone and mineralizing posterior face of the cusp. Plume formation coincides with the appearance of iron in the superior epithelium and the onset of mineralization at tooth row 13. The plume persists during the delivery of phosphorous and calcium into the tooth core, and is the final region of the cusp to become mineralized. The presence of the stylus canal was confirmed in a further 18 chiton species, revealing that the canal is common to polyplacophoran molluscs. These new data strongly support the growing body of evidence highlighting the importance of the junction zone for tooth mineralization in chiton teeth, and indicate that the chemical and structural environment within the tooth cusp is under far greater biological control than previously considered.

The effect of induced biphasic pulsed currents on re-epithelialization of a novel wound healing model.

The coordinated migration of keratinocytes is crucial to cutaneous wound healing; failure of keratinocytes to migrate into a wound can lead to chronic non-healing wounds. Keratinocyte migration can be influenced by applied electrical fields. Our aim was to investigate whether keratinocyte migration could be accelerated by applying an induced biphasic pulsed electrical field. We developed two in vitro biological systems models for this purpose: a keratinocyte colony-forming model and a reconstituted skin wound healing model with biphasic pulsed currents. Our in vitro skin models were capable of generating trans-epithelial potentials (TEP) similar to in vivo mammalian skin. Histological examination of the wound healing model also indicated that re-epithelialization occurred in a similar manner to that seen in vivo, although no evidence of a reconstitution of a basement membrane was seen during the 14 days in vitro experimental period. We found that growth of keratinocyte colonies and keratinocyte migration in an in vitro wound bed were not significantly affected by induced short duration biphasic pulsed currents at a frequency of 0.5 Hz of 100 and 200 mV/mm.

The phenotype of in vitro reconstituted normal human oral epithelium is essentially determined by culture medium.

OBJECTIVE: To evaluate the role of various culture media and serum supplement on growth of oral cells in monolayer, and on morphogenesis of in vitro reconstituted normal human oral epithelium. METHODS: Primary keratinocytes and fibroblasts were isolated from normal human buccal mucosa. The monolayers were assessed by growth curve analysis and morphology. The organotypic cultures were evaluated by morphometry, immunohistochemistry, and TUNEL. RESULTS: FAD medium (a 3:1 mixture of Dulbecco's modified Eagle's medium and Ham's F12 medium) was able to support fibroblast growth in defined conditions, and to diminish the negative effect of physiological Ca concentration on keratinocytes in monolayers. Medium type had a profound influence on morphogenesis of in vitro reconstituted human oral epithelium. FAD medium was superior to other types of medium tested in supporting both epithelial growth and differentiation. Defined conditions supported epithelial morphogenesis equally well as serum-containing medium. CONCLUSIONS: This study points to an essential role of medium composition for optimized growth and differentiation of primary organotypic cultures.

Effects of adding extra molasses to a texturized calf starter on rumen development, growth characteristics, and blood parameters in neonatal dairy calves.

A texturized calf starter containing 5 (control) or 12% molasses [on a dry matter (DM) basis] was fed to dairy calves to determine effects on intake, growth, blood parameters, and rumen development. Forty-six Holstein calves (26 male and 20 female) were started at 2 +/- 1 d of age and studied for 42 d. Starter DM intake was measured and fecal scoring was conducted daily. Growth and blood parameter measurements were conducted weekly. A subset of 6 male calves (3 per treatment) was euthanized at 4 wk of age, and rumen tissue sampled for rumen epithelial growth measurements. Starter sugar content was significantly increased in the starter containing extra molasses. Postweaning and overall starter DM intake, overall total DM intake, daily heart girth change, and final heart girth were significantly decreased, whereas overall average daily gain tended to decrease when calves received starter containing 12% molasses. However, blood volatile fatty acid concentrations were significantly increased when calves received a starter containing 12% molasses. No significant differences were observed between calves receiving starters containing 5 or 12% molasses for all other variables. The data indicates that adding extra molasses to a texturized calf starter decreases intake and structural growth, possibly causing decreased weight gain, but increases blood volatile fatty acid concentrations and slightly increases ruminal development. However, feed handling and physical prehension problems in addition to the negative influences on calf growth and intake do not support increasing starter molasses content to 12% of the supplement.

Restoration of thymocyte development and function in zap-70-/- mice by the Syk protein tyrosine kinase.

The Syk family of protein tyrosine kinases, consisting of ZAP-70 and Syk, associate with the pre- and alphabeta T cell antigen receptors (TCRs) and undergo tyrosine phosphorylation and activation following receptor engagement. Thymocyte development in zap-70-/- mice is blocked at the CD4+CD8+ TCR(lo) stage. The presence of Syk in the thymus has raised the possibility that Syk may be able to mediate TCR function. To determine if Syk can play a role in thymocyte development, we generated zap-70-/- mice expressing a human syk cDNA. Syk expression restored both thymocyte development and function. In addition, Syk function required the CD45 transmembrane protein tyrosine phosphatase. Hence, ZAP-70 and Syk can play overlapping functions and exhibit similar regulatory mechanisms in mediating alphabeta T cell development.

Differentially regulated epithelial expression of an Eph family tyrosine kinase (fHek2) during tracheal surface airway and submucosal gland development.

A ferret model was used to evaluate the potential role of an Eph family tyrosine kinase (fHek2) in tracheal development of surface airway epithelium and submucosal glands. A partial 2.6-kb cDNA fragment of fHek2 was isolated from a ferret tracheal/lung cDNA library. Sequence analysis demonstrated that this gene is the ortholog to the previously cloned human Hek2 gene. In situ hybridization analysis of fHek2 mRNA expression on ferret tracheal developmental time points revealed an expression pattern within a subset of surface airway epithelial cells which remained relatively constant throughout tracheal development (from -2 d in utero to adult). In contrast, developing tracheal submucosal glands at 3-day postnatal time points demonstrated little fHek2 mRNA expression. However, expression of fHek2 significantly increased more than 4-fold over the course of gland development to adulthood. These findings, which demonstrate a uniquely regulated pattern of fHek2 mRNA expression between surface airway epithelium and submucosal glands, have implications on regulatory processes which control differentiation and/or maturation of secretory structures in the lung. Such findings may be useful in further delineating the mechanisms which control cellular differentiation in the lung and how these processes are abnormally regulated in hypersecretory diseases such as chronic bronchitis, asthma, and cystic fibrosis.

Inhibition of hypoxic pulmonary hypertension by heparins of differing in vitro antiproliferative potency.

Heparin inhibits smooth-muscle cell (SMC) growth in vitro and inhibits the development of hypoxic pulmonary hypertension and vascular remodeling in vivo. We wondered whether preparations of heparin with different antiproliferative potency in vitro would differ in their ability to inhibit the development of hypoxic pulmonary hypertension in vivo. Two such heparins, a weakly antiproliferative lot of Elkins-Sinn (E-S) (% inhibition of SMC growth at 10 micrograms/ml = 13 +/- 9% [mean +/- SEM, n = 24]) and a more active lot from Upjohn (UJ) (% inhibition = 71 +/- 12% [n = 12, p < 0.05 versus E-S]), were infused subcutaneously (300 U.S.P. units/day; E-S 300 versus UJ 300) via an osmotic pump into guinea pigs exposed to hypoxia (10% O2) for 10 d, after which pulmonary artery pressure (PAP; mm Hg) and cardiac index (CI; ml/min/kg) were measured in room air. Hypoxic controls (HC) received saline. PAP increased from 11 +/- 1 mm Hg in normoxic controls (NC) (n = 5) to 24 +/- 1 mm Hg in HC (n = 8, p < 0.05). The PAP was lower in the E-S 300 (21 +/- 1; n = 7, p < 0.05 versus HC and NC) and even lower in the UJ 300-treated group (18 +/- 0.5; n = 7, p < 0.05 versus HC and NC). Total pulmonary vascular resistance (TPR; mm Hg/ml/min/kg) increased significantly from 0.038 +/- 0.002 in NC to 0.076 +/- 0.003 (p < 0.05) in HC. There was no difference in TPR between the HC and the E-S 300-treated group.(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of preclosure colostrum intake on the development of the intestinal epithelium of artificially reared piglets.

Trophic factors in mammalian colostrum promote the growth of the small intestine of neonates. To investigate the effect of colostrum feeding on the expression of specific intestinal proteins, animals were reared in a minimal disease unit and fed either sow colostrum or a commercial substitute by gastric intubation at 3-hour intervals over the first 24 h of life. Animals were then reared on a commercial milk replacer and fed over a maximum period of 5 weeks. Intestinal protein, DNA and histology data suggested a positive effect of colostrum on intestinal growth in the initial postnatal period. At week 1 post partum intestinal lactase was found to decline significantly in colostrum-fed (CF) piglets compared to substitute-fed animals. This effect was no longer apparent at 3 and 5 weeks post partum. Sucrase activity was significantly greater in CF piglets and this effect was sustained during the 5 postpartum weeks studied. The changes in enzyme activity could be correlated with posttranslational sialylation of intestinal membranes. These result suggest that feeding colostrum enhances the maturational decline in lactase activity and the expression of sucrase activity. The role of glycosylation of enzyme proteins in relation to their biological activity is discussed.

Multiple graft harvestings from deep partial-thickness scald wounds healed under the influence of weak direct current.

The time required for wound healing, contraction, and hypertrophic scarring often limit the use of deep partial-thickness burn wounds as donor sites for split-thickness grafts. We have examined the effects of weak direct current and silver nylon dressings on the healing of partial-thickness scald burns, split-thickness grafts taken from these wounds when healed, and the resulting donor sites in a guinea pig model. Dorsal scald wounds treated with weak direct current reepithealized by 12 days postinjury. Split-thickness grafts taken from healed scald wounds showed more rapid revascularization with direct current treatment than did control grafts. Grafts and donor sites treated with direct current showed more rapid reepithelialization, decreased contraction, improved hair survival, and decreased dermal fibrosis when compared to controls not treated with direct current. Only donor wounds treated with weak direct current were reusable as donor sites.

Density-dependent regulation of growth of BSC-1 cells in cell culture: control of growth by low molecular weight nutrients.

BSC-1 cells, epithelial cells of African green monkey kidney origin, show pronounced density-dependent regulation of growth in cell culture. Growth of the cells is rapid to a density of approximately 1.5 x 10(5) cells/per cm(2) in Dulbecco-modified Eagle's medium supplemented with 10% calf serum. Above this "saturation density," growth is much slower. It has been found that the glucose concentration in the culture medium is important in determining the "saturation density." If the glucose concentration is increased 4-fold, the "saturation density" increases approximately 50%. Reduction of the "saturation density" of BSC-1 cells is also possible by decreasing the concentrations of low molecular weight nutrients in the culture medium. In medium supplemented with 0.1% calf serum, decreasing the concentrations of all of the organic constituents of the medium, from the high levels present in Dulbecco-modified Eagle's medium to concentrations near physiological levels, decreases the "saturation density" by approximately half. The decreased "saturation density" is not the result of lowering the concentration of any single nutrient but rather results from reduction of the concentrations of several nutrients. When the growth of BSC-1 cells is limited by low concentrations of all of the nutrients, some stimulation of growth results from increasing, separately, the concentrations of individual groups of nutrients, but the best growth stimulation is obtained by increasing the concentrations of all of the nutrients. The "wound healing" phenomenon, one manifestation of density-dependent regulation of growth in cell culture, is abolished by lowering the concentration of glutamine in the medium. Density-dependent regulation of growth of BSC-1 cells in cell culture thus appears to be a complex phenomenon that involves an interaction of nutrient concentrations with other regulatory factors.