Standardization of platelet parameters determination in blood samples collected with EDTA

The Abbott Cell-Dyn 3000 automated hematological analyzer prepares a histogram of platelet volume, which is measured by a technique using electrical impedance, inferring from its platelet count (PLT), mean platelet volume (MPV) and platelet distribution width (PDW). This equipment also calculates the plateletcrit (PCT). These platelet parameters may be important to evaluate platelet function, but they require standardization, because platelets swell when in contact with ethylenediaminetetra-acetic acid salts, hence an increase in blood sample storage time produces artificially increased results. To assess the effect of storage time on MPV, PLT, PDW and PCT, blood samples from 23 sickle cell anemia patients during steady state (Group I) and 50 from healthy controls (Group II) were placed in Vacutainer® tubes with dipotassium ethylenediaminetetra-acetic acid and measured over a period of 1440 minutes (24 h) at the following times: immediately after the venipuncture (time 0), 15, 30, 60, 120, 240, 360, 480 and 1440 minutes. The mean values of MPV and PCT were significantly increased (p<0.00001) along the storage time in both groups. The mean values of PLT and PDW were practically stable (p>0.05) throughout the storage time in both groups.

Effects of storage time on incubating egg gas pressure, thyroid hormones, and corticosterone levels in embryos and on their hatching parameters

Incubating eggs (1,800 total) produced by a commercial flock of Cobb broiler breeders were used to determine the effects of storage duration (3 and 18 d) on gas partial pressure, thyroid hormones, and hatching parameters. Partial pressure of oxygen (pO2) and carbon dioxide (pCO2) were measured on d 18 and at internal pipping (IP) during incubation. Blood samples were collected for determination of triiodothyronine (T3), thyroxine (T4), and corticosterone concentrations in the embryos at IP and in newly hatched chicks. From 464 to 510 h of incubation, eggs were checked individually every 2 h to determine the timing and duration of IP, external pipping (EP), and total hatching time. At 18 d of incubation and at IP, pCO2 was greater in air cell of eggs stored for 3 d compared to those stored for 18 d (P < 0.05), but pO2 was greater in eggs stored for 18 d. At IP, T3 and corticosterone levels were higher in plasma of the embryos of eggs stored for 3 d compared to those stored for 18 d, but it was the reverse in newly hatched chicks (P < 0.05). Embryos from eggs stored for 18 d required more time to complete IP compared to embryos of eggs stored for only 3 d (P < 0.05), whereas the duration of EP was not affected by storage. The overall longer incubation was, however, not only due to prolonged IP but also to later occurrence of IP. It was concluded that prolonged IP as a result of long storage may be related to the late increase in corticosterone level, which may be a necessary stimulus for higher T 3/T4 ratio, late increase in pCO2 level, and decrease in pO2. The effect of long storage was a delay in hatching and a continuous increase in T3 due to higher corticosterone levels between IP and hatching, which may be an indication of the more stressful event of hatching of embryos from eggs stored longer. Differences in pCO2, pO2, T3, T4, and corticosterone levels in the incubating eggs may be manifestations of these changes culminating in altered hatching parameters and consequently differences in chick quality and growth potentials.

Influence of the Light-Curing Unit, Storage Time and Shade of a Dental Composite Resin on the Fluorescence

The aim of this study was to determine the influence of three light-curing units, storage times and colors of the dental composite resin on the fluorescence. The specimens (diameter 10.0 +/- 0.1 mm, thickness 1.0 +/- 0.1 mm) were made using a stainless steel mold. The mold was filled with the microhybrid composite resin and a polyethylene film covered each side of the mold. After this, a glass slide was placed on the top of the mold. To standardize the top surface of the specimens a circular weight (1 kg) with an orifice to pass the light tip of the LCU was placed on the top surface and photo-activated during 40 s. Five specimens were made for each group. The groups were divided into 9 groups following the LCUs (one QTH and two LEDs), storage times (immediately after curing, 24 hours, 7 and 30 days) and colors (shades: A(2)E, A(2)D, and TC) of the composite resin. After photo-activation, the specimens were storage in artificial saliva during the storage times proposed to each group at 37 C and 100% humidity. The analysis of variance (ANOVA) and Tukey's post-hoc tests showed no significant difference between storage times (immediately, 24 hours and 30 days) (P > 0.05). The means of fluorescence had difference significant to color and light-curing unit used to all period of storage (P < 0.05). The colors had difference significant between them (shades: A2D < A2E < TC) (P < 0.05). The Ultraled (LED) and Ultralux (QTH) when used the TC shade showed higher than Radii (LED), however to A2E shade and A2D shade any difference were found (P > 0.05).

Effects of egg storage time on spread of hatch, chick quality, and chick juvenile growth

A total of 1,800 incubating eggs produced by a commercial flock of Cobb broiler breeders was used to determine the effects of storage duration (3 or 18 d) on spread of hatch and chick quality. Chick relative growth (RG) at the end of 7 d of rearing was also determined as a measure of the chick performance. Chick quality was defined to encompass several qualitative characteristics and scored according to their importance. Eggs stored for 3 d hatched earlier than those stored for 18 d (P < 0.05). Hatching was normally distributed in both categories of eggs, and the spread of hatch was not affected by storage time (P = 0.69). Storage duration of 18 d reduced the percentage of day-old chick with high quality as well as average chick quality score (P < 0.05). RG varied with length of egg storage, quality of day-old chick, and the incubation duration (P < 0.05). Eighteen-day storage of eggs not only resulted in longer incubation duration and lower quality score but also depressed RG. Chick quality as defined in this study was correlated to RG and storage time. It was concluded that day-old chick quality may be a relatively good indicator of broiler performance. The results suggest however that in order to improve performance prediction power of chick quality, it would be better to define it as a combination of several qualitative aspects of the day-old chick and the juvenile growth to 7 d.

Microhardness of glass ionomer cements indicated for the ART technique according to surface protection treatment and storage time

The aim of this study was to assess the microhardness of 5 glass ionomer cements (GIC) - Vidrion R (V, SS White), Fuji IX (F, GC Corp.), Magic Glass ART (MG, Vigodent), Maxxion R (MR, FGM) and ChemFlex (CF, Dentsply) - in the presence or absence of a surface protection treatment, and after different storage periods. For each GIC, 36 test specimens were made, divided into 3 groups according to the surface protection treatment applied - no protection, varnish or nail varnish. The specimens were stored in distilled water for 24 h, 7 and 30 days and the microhardness tests were performed at these times. The data obtained were submitted to the ANOVA for repeated measures and Tukey tests (α = 5%). The results revealed that the mean microhardness values of the GICs were, in decreasing order, as follows: F > CF = MR > MG > V; that surface protection was significant for MR, at 24 h, without protection (64.2 ± 3.6a), protected with GIC varnish (59.6 ± 3.4b) and protected with nail varnish (62.7 ± 2.8ab); for F, at 7 days, without protection (97.8 ± 3.7ab), protected with varnish (95.9 ± 3.2b) and protected with nail varnish (100.8 ± 3.4a); and at 30 days, for F, without protection (98.8 ± 2.6b), protected with varnish (103.3 ± 4.4a) and protected with nail varnish (101 ± 4.1ab) and, for V, without protection (46 ± 1.3b), protected with varnish (49.6 ± 1.7ab) and protected with nail varnish (51.1 ± 2.6a). The increase in storage time produced an increase in microhardness. It was concluded that the different GICs, surface protection treatments and storage times could alter the microhardness values.

Effects of drying rate and storage time on Magnolia ovata Spreng. seed viability

Magnolia ovata seeds have been reported as desiccation sensitive. In order to test if the drying rate would affect the assessment of storage behaviour of these seeds, the effect of different drying rates and storage times on the viability was tested. Seeds were dried over activated silica gel (fast drying) or salt solutions for different periods (slow drying) and stored at -20°C. Partial drying transiently increased the final germination and the germination speed index, but further drying resulted in reduction of these parameters. Drying rate affected the final germination and vigour. Seeds that were slow-dried to 0.10 g H 2O ̇ g -1 dw retained high viability when compared with seeds desiccated to the same water content level by the fast drying method, although their vigour was reduced. Only slow-dried seeds could be stored at -20°C for 90 d without reduction of viability. These data suggested that the storage behaviour of seeds of M. ovata seeds should be classified as intermediate.

The Effects of Refrigeration Temperature and Storage Time on Apoptotic Markers in Equine Semen

Evaluation of the damage caused by the sperm preservation process is crucial to improving fertilization rates. The objective of this study was to evaluate the effects of refrigeration temperature (5°C and 15°C) and storage time (0, 12, 24, 48, and 72 hours) on apoptotic markers in equine semen. Membrane phosphatidylserine translocation index, caspase activation index, and DNA fragmentation index were analyzed using epifluorescence microscopy. Analysis of variance was used for statistical analysis, and Tukey test was used to compare means. The significance level was set at P < .05. The results demonstrated that for transport duration shorter than 24 hours, semen quality was maintained when stored at either 5°C or 15°C. A storage temperature of 5°C should be used when it is necessary to transport semen for longer than 24 hours. There was a significant decrease in semen quality after 48 hours of refrigeration. © 2013 Elsevier Inc.

Effect of Storage Time and Temperature of Equine Epididymis on the Viability, Motion Parameters, and Freezability of Epididymal Sperm

The recovery of sperm from the epididymal cauda may be the last chance to obtain genetic material when sudden death or serious injuries occur in valuable stallions. However, the lack of technical knowledge regarding the storage and transportation of the epididymis often prevents the preservation of the sperm. Therefore, the aim of this study was to compare sperm parameters of sperm obtained immediately after orchiectomy with sperm recovered from epididymal cauda at different times after storage at 5°C and at room temperature (RT). For that, 48 stallions of different breeds were used. In group 1 (control group), eight stallions were used, and the harvest of the epididymal sperm was performed immediately after orchiectomy. In group 2, 40 stallions were used, which were divided into five groups according to the storage time of the epididymis after orchiectomy (6, 12, 18, 24, or 30 hours), making a total of eight stallions per group. One epididymis of each stallion was stored at 5°C, and the contralateral epididymis was stored at RT, both for the same period. The sperm parameters of total motility, progressive motility, progressive linear velocity, curvilinear velocity, percentage of rapid sperm, and plasma membrane integrity were evaluated in all the groups after sperm recovery, resuspension in a sperm freezing diluent, and thawing. In conclusion, the storage of the testis-epididymis complex at 5°C provided better preservation of epididymal sperm than the storage at RT, and regardless of the temperature, the progressive motility is the sperm parameter that is most sensitive to storage time. © 2013 Elsevier Inc.

Effect of Egg Rearing Temperature and Storage Time on the Biological Characteristics of the Predatory Stink Bug Podisus nigrispinus (Hemiptera: Pentatomidae)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

A castor oil-containing dental luting agent: effects of cyclic loading and storage time on flexural strength

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

The effects of ice-water storage on blood gas and acid-base measurements

Objective: To determine the effects of storage of arterial and venous blood samples in ice water on blood gas and acid-base measurements.Design: Prospective, in vitro, laboratory study.Setting: School of veterinary medicine.Subjects: Six healthy dogs.Measurements and main results: Baseline measurements of partial pressure of oxygen (PO2), partial pressure of carbon dioxide (PCO2), pH, hemoglobin concentration (tHb), oxyhemoglobin saturation, and oxygen content (ContO(2)) were made. Bicarbonate (HCO3) and standard base excess (SBE) were calculated. Arterial and venous blood samples were separated into 1 and 3 mL samples, anaerobically transferred into 3 mL plastic syringes, and stored in ice water for 6 hours. Measurements were repeated at 15, 30 minutes, and 1, 2, 4, and 6 hours after baseline measurements. Arterial (a) PO2 increased significantly from baseline after 30 minutes of storage in the 1 mL samples and after 2 hours in the 3 mL samples. Venous (v) PO2 was significantly increased from baseline after 4 hours in the 1 mL samples and after 6 hours in the 3 mL samples. The pHa significantly decreased after 2 hours of storage in the 1 mL samples and after 4 hours in the 3 mL samples. In both the 1 and 3 mL samples, pHv decreased significantly only after 6 hours. Neither the arterial nor the venous PCO2 values changed significantly in the 1 mL samples and increased only after 6 hours in the 3 mL samples. No significant changes in tHb, ContO(2), SBE, or HCO3 were detected.Conclusions: the PO2 of arterial and venous blood increased significantly when samples were stored in plastic syringes in ice water. These increases are attributable to the diffusion of oxygen from and through the plastic of the syringe into the blood, which occurred at a rate that exceeded metabolic consumption of oxygen by the nucleated cells.

Electrical conductivity of soybean seeds after storage in several environments

Membrane integrity, as measured by electrical conductivity (EC), is suggested as an indicator of seed vigor in soybean [Glycine max (L.) Merrill] seeds. This study evaluated the effect of storage time and temperature on EC of six soybean seed lots (two lots each of high, medium and low vigor). All seed lots were adjusted to 120 g kg(-1) seed moisture, sealed in aluminum foil packets and placed in storage at 10 and 20 degreesC or stored unsealed in multi-wall paper bags in warehouse (WH) conditions at Lexington, KY, USA for 486 days. Four of the six seed lots were also stored unsealed at 10 degreesC. All seed lots were sampled at 3-month intervals and evaluated for seed moisture (SMC), standard germination (SG) and vigor [accelerated aging (AA) and EC]. After 91 and 204 days in storage, samples initially stored at 20 degreesC and WH were moved to 10 degreesC and sampled at the same intervals. Seed moisture content for unsealed samples equilibrated at 107 g kg(-1) (+/-9 g kg(-1)) in both the WH and 10 degreesC environments. No change in SG occurred for seeds stored sealed (120 g kg(-1)) at 10 degreesC, except for the low vigor seed lots which declined significantly at the last sample date. The AA germination declined significantly for all, seed lots stored sealed at 10 degreesC, however the EC did not change during the same storage period. Seeds stored sealed at 20 degreesC and unsealed in the WH showed rapid declines in AA and SG and significant increases in EC. When these seeds were moved to 10 degreesC, however, the AA continued to decline while the EC remained at the same level (no significant change) for the remainder of the seed storage period. Thus whilst the AA declined in all environments, the EC only increased at higher temperatures (20 degreesC, WH) but showed little change during storage at 10 degreesC. Thus, precautions must be taken if using EC to measure soybean seed vigor following storage at 10 degreesC.

Effect of water storage and heat treatment on the cytotoxicity of soft liners

Objective: To evaluate the effect of water storage time on the cytotoxicity of soft liners.Methods: Sample discs of soft liners Dentusoft, Dentuflex, Trusoft, Ufi-Gel-P and denture base acrylic resin Lucitone-550 were prepared and divided into four groups: GN: No treatment, G24: Stored in water at 37 degrees C for 24 h; G48: Stored in water at 37 degrees C for 48 h, GHW: Immersed in water at 55 degrees C for 10 min. To analyse the cytotoxic effect, three samples of each group were placed in tubes with Dubelcco's Modified Eagle Mediums and incubated at 37 degrees C for 24 h. During this period, the toxic substances were leached to the culture medium. The cytotoxicity was analysed quantitatively by the incorporation of radioactivity H-3-thymidine checking the number of viable cells (synthesis of DNA). The data were statistically analysed using two-way ANOVA and Tukey's honestly significant difference tests (alpha = 0.05).Results: Treatments did not reduce the cytotoxicity effect of the soft liners (p > 0.05). It was found that Ufi-Gel-P had a non-cytotoxic effect, Trusoft had a slightly cytotoxic effect, Dentuflex had a moderated cytotoxic effect, Dentusoft alternated between slightly and non-cytotoxic effect, and Lucitone-550 had non-cytotoxic effect when stored in water for 48 h.Conclusion: The effect of water storage and the heat treatment did not reduce the cytotoxicity of the soft liners.

LH and FSH concentration and follicular development in Nellore heifers submitted to fixed-time artificial insemination protocols with different progesterone concentrations

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