Expression of cell cycle regulatory proteins in epithelial components of dental follicles

Dental follicle is a component of tooth germs, which remain adjacent to the crown of unerupted or impacted teeth. Under the influence of pathologic changes, however, dental follicles that possess reduced epithelium can proliferate into stratified squamous epithelium as far as originate dental cysts. In order to clarify the role of apoptosis and cellular proliferation herein, expression of p53 and PCNA was examined in epithelial components of dental follicles associated with impacted third molars by means of immunohistochemistry. A total of 40 cases was included in this study being 22 cases with reduced epithelium and 18 cases with stratified epithelium. Expression of p53 expression was weak or not detected in dental follicles with reduced and stratified squamous epithelium. By contrast, PCNA positive cells were evidenced in basal and supra basal layers of the stratified squamous epithelium and in reduced epithelium of dental follicles, but without any significant statistically differences between them (P > 0.05). In conclusion, these data suggest that dental follicles possess proliferative activity as depicted by PCNA-positive nuclei in some epithelial cells. However, the biological behavior of dental follicles during the late stage of dental eruptive process may not be associated with deregulation of death and/or cell proliferation.

Toxicogenomic activity of gemcitabine in two TP53-mutated bladder cancer cell lines: special focus on cell cycle-related genes

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Cell cycle arrest and apoptosis in TP53 subtypes of bladder carcinoma cell lines treated with cisplatin and gemcitabine

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Expression of genes related to apoptosis, cell cycle and signaling pathways are independent of TP53 status in urinary bladder cancer cells

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Effects of dietary trace mineral sources and levels fed to layers in their second laying cycle on the quality of eggs stored at different temperatures and for different periods

This study aimed at evaluating the effects of trace mineral levels and sources supplemented to diets fed to semi-heavy layers in their second laying cycle on the quality of eggs stored for 14 days at different temperatures. The experimental diets consisted of the inclusion of inorganic trace minerals (T1 - control: 100% ITM) and five supplementation levels of organic trace minerals (carboaminophopho chelates) (110, 100, 90, 80, and 70% OTM). Trace mineral inclusion levels (mg/kg feed) were: T1: control - 100% ITM: Zn (54), Fe (54), Mn (72), Cu (10), I (0.61) Se (0.3); T2 - 110% OTM: Zn (59.4), Fe (59.4), Mn (79.2), Cu (11.88), I (1.21) Se (0.59); T3 - 100%: OTM: Zn (54), Fe (54), Mn (72), Cu (10.8), I (1.10) Se (0.54); T4 - 90% OTM: Zn (48.6), Fe (48.6), Mn (64.8), Cu (9.72), I (0.99) Se (0.49); T5 - 80% OTM: Zn (43.2), Fe (43.2), Mn (57.6), Cu (8.64), I (0.88), Se (0.43); T6 - 70% OTM: Zn (37.8), Fe (37.8), Mn (50.4), Cu (7.56), I (0.77) Se (0.38). A completely randomized experimental design in a split-plot arrangement with 60 treatments of four replicates each was applied. The combination of six diets versus storage temperature (room or under refrigeration) was randomized in plots, whereas the sub-plots consisted of storage times (0, 3, 7, 10, and 14 days). Data were submitted to analysis of variance of a model in slip-plots in time using the software package SAS (2000) at 5% probability level. It was concluded that 70% OTM supplementation can be used with no damage to egg quality, independently from storage temperature or time. The quality of refrigerated eggs stored up to 14 days is better than those stored at room temperature.

Effect of organic mineral supplementation on the egg quality of semi-heavy layers in their second cycle of lay

This study was carried out to evaluate the effects of dietary trace mineral levels and sources on egg quality parameters of second-cycle semi-heavy layers. A number of 360 72-week-old layers were submitted to forced molting. Upon return of lay (83 weeks of age), birds were distributed according to a completely randomized experimental design of six treatments with six replicates of 10 birds each. The control treatment consisted of 0.10% dietary supplementation of trace minerals from inorganic sources, which was proportionally replaced by five levels (110, 100, 90, 80, 70%) of an organic trace mineral supplement containing 30, 30, 40, 6, 0.61, and 0.3 g/kg product of Zn, Fe, Mn, Cu, I, and Se, respectively. All diets contained equal protein, energy, and amino acid levels. Every 28 days of the experimental period (112 days) four eggs per replicate were collected for egg quality evaluation. The following parameters were evaluated: specific gravity, yolk, albumen and eggshell percentages, yolk index, Haugh units, and eggshell thickness and breaking strength. One sample per replicate, consisting of the pool of the yolks of three eggs collected at the end of each experimental period, was used to assess protein and mineral (Ca, P, Cu, Fe, Mn, and Zn) contents. The results were submitted to ANOVA, and means to the test of Tukey at 5% significance level. The evaluated trace mineral levels and sources did not influence any of the studied egg quality parameters. It was concluded that reducing organic trace mineral supplementation in up to 70% relative to 100% inorganic trace mineral supplementation does not affect egg parameters and therefore, can be applied to the diet of semi-heavy layers in their second cycle of lay.

Calcium levels and limestone particle size in the diet of commercial layers at the end of the first production cycle

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Calcium and available phosphorus levels for laying hens in second production cycle

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Effect of the calcium level and limestone particle size on the performance of semi-heavy layers in the second cycle of egg production

An experiment was carried out at the Research and Development Unit of Brotas aiming at evaluating dietary calcium level and limestone particle size on the production performance of commercial (Hy-Line Brown) layers in the second lay cycle. Experiment duration was 112 days. A total number of 288 hens, with 83 weeks of age in the beginning of the experiment, were used in a completely randomized experimental design in a factorial arrangement of 2x3, with two calcium levels (3.5 and 4.0%) and three limestone particle size compositions: 100% fine limestone (FL), 30% coarse limestone (CL) + 70% fine limestone (FL), and 50% (CL) + 50% (FL), with six replicates of eight birds each. Egg weight (g), egg production (%), egg mass (%), feed intake (g), feed conversion ratio (kg/dz and kg/kg), mortality (%), and egg loss (%) were evaluated. The analysis of variance did not detect significant differences (p>0.05) among treatments on any of the evaluated performance parameters. It was concluded that the tested calcium levels and limestone particle composition did not influence the performance of semi-heavy layers in second production cycle.

Forced-Molting methods and their effects on the performance and egg quality of japanese quails (Coturnix japonica) in the second laying cycle

The experiment was carried out in the experimental poultry house of the Research and Development Unit of Brotas of Agência Paulista de Tecnologia dos Agronegócios do Centro-Oeste, SP, Brazil. The objective of the study was to evaluate the performance of Japanese quails submitted to forced molting aiming at optimizing the use of the same quail flock by promoting a second laying cycle. A total number of 400 67-day-old Japanese quails in lay, previously submitted to 14 days of forced molting, was distributed in a completely randomized experimental design into five treatments (T1= not submitted to forced molting, T2= 03 days of fasting + fed ad libitum, T3= 01 days of fasting + 13 days of feed restriction, T4= 02 days of fasting + 12 days of feed restriction, and T5= 03 days of fasting + 11 days of feed restriction. Feeds were contained equal nutrient levels, and were formulated according to NRC (1994) recommendations. There were significant differences among the studied treatments. Although the treatment of 3 days of fasting followed by ad libitum feeding resulted in lower egg weight, it promoted better lay percentage, egg mass, and feed conversion ratios (FCR/dz and FCR/kg). on the other hand, 3 days of fasting followed by restricted feeding resulted in higher feed intake and worse feed conversion ratios (FCR/dz and FCR/kg). When birds were not submitted to forced molting, they presented lower lay percentage and egg mass.

Viability and cell cycle analysis of equine fibroblasts cultured in vitro

This experiment aimed to study equine fibroblasts in culture analyzing and the cell cycle and viability of cells pre- and post-freezing. Skin fragments were obtained from 6 horses and cultured in DMEM high glucose + 10% FCS in 5% CO(2) until the beginning of confluence. Two passages were performed before freezing. Cells subjected to serum starvation (0.5% FCS) were analyzed for viability and cell cycle at 24, 48, 72, 96, 120, 144 and 168 h of culture. For the confluent groups, cells were analyzed at the moment they achieved confluence. Cellular viability was assisted with Hoescht 33342 and propidium iodide. The analysis of apoptosis/necrosis and cell cycle was performed using a flow cytometer (FACS Calibur BD(A (R))) after staining the cells with annexin V and propidium iodide. Both optical microscopy and flow cytometry confirmed that cellular viability was similar for serum starvation and confluent groups (average 84%). Similarly, both methods were efficient to synchronize the cell cycle before freezing. However, after thawing, serum starvation, for more than 24 h, was superior to culture for synchronizing cells in G0/G1 (69% x 90%). The results of this experiment indicate that equine fibroblasts can be efficiently cultured after thawing.

Immunohistochemical detection of receptors for oestrogen and progesterone in endometrial glands and stroma during the oestrous cycle in Nelore (Bos taurus indicus) cows

The aim of the present study was to monitor endometrial distribution and concentrations of oestrogen receptors alpha (ER alpha) and progesterone receptors (PR) by immunohistochemistry in Nelore cows (Bos taurus indicus) during the oestrous cycle. Blood samples were collected for progesterone measurement and endometrial samples were taken from the uterine horn contra lateral to the corpus luteum in 16 cows at days 0 (ovulation), 5, 9, 13 and 19 of the oestrous cycle. Immunostaining evaluation for ER alpha and PR in the glandular epithelium and uterine stroma was performed by two methods: positive nuclei counting and staining intensity of the nuclei. Specific positive staining reactions for both receptors were limited to cell nuclei and they were not identified in the cytoplasm. The proportion of ER alpha positive nuclei had a temporal variation throughout the oestrous cycle in both cell types evaluated and was higher in uterine stroma than the glandular epithelium (p < 0.05). The greatest proportion of ER alpha stained nuclei was observed at oestrus and during the initial and mid luteal phase (days 5, 9 and 13) (p < 0.05) in the glandular epithelium and at days 0, 5 and 9 in the uterine stroma (p < 0.01). The proportion of PR positive nuclei remained constant throughout the entire oestrous cycle for both cell types evaluated (p > 0.05). A higher proportion of PR positive nuclei was measured in the uterine stroma compared with the glandular epithelium (p < 0.05). Intensity of staining for ER alpha and PR varied throughout the oestrous cycle (p < 0.01). There was a higher staining intensity at days 0 and 5 in the stroma for ER alpha (p < 0.01) and PR (p < 0.01) and in the glandular epithelium at days 0, 5, 9 and 13 for ER alpha (p < 0.01) and at days 0, 5 and 9 for PR (p < 0.01) when compared with the other evaluated days. These data demonstrate that ER alpha and PR expression varied throughout the oestrous cycle in Nelore cows, in general with highest concentrations at oestrus and the lowest during the luteal phase. This is similar to patterns observed in Bos taurus taurus.

Follicular Development in Ewes During Natural and Prostaglandins Induced Estrous Cycle

The follicular development was evaluated in ovine females during natural and prostaglandin-F(2 alpha) (PG) induced estrous cycle. Ewes were randomly divided in two treatments (n=7/treatment): T1 with natural cycle and T2 synchronized with two injections of PG. From one day before PG injection until next ovulation, daily transrectal ultrasonography was done. All follicles >= 2 mm were assessed. During the interovulatory intervals, follicular growth and regression occurred in a wave like pattern (2-3 waves). The maximum diameter of the largest follicle of the first wave was greater in T1 (5.83 +/- 0.31 mm) compared with T2 (5.0 +/- 0.1 mm; P<0.01), but there was no significant difference among the emergency day of largest follicle, during the growth phase of the follicular waves. The duration of the plateau phase in wave 2 differed between the two treatments (P<0.05) showing 0.83 +/- 0.31 and 1.83 +/- 0.17 d, for natural and synchronized treatment, respectively. Growth rate did not differ between treatments. Presence of new luteal tissue was detected on day 3 after ovulation. In conclusion, the follicular development was similar in female ovine during natural and PG induced estrous cycle.