Preparation and characterization of cockle shell aragonite nanocomposite porous 3D scaffolds for bone repair

The demands for applicable tissue-engineered scaffolds that can be used to repair load-bearing segmental bone defects (SBDs) is vital and in increasing demand. In this study, seven different combinations of 3 dimensional (3D) novel nanocomposite porous structured scaffolds were fabricated to rebuild SBDs using an extraordinary blend of cockle shells (CaCo₃) nanoparticles (CCN), gelatin, dextran and dextrin to structure an ideal bone scaffold with adequate degradation rate using the Freeze Drying Method (FDM) and labeled as 5211, 5400, 6211, 6300, 7101, 7200 and 8100. The micron sized cockle shells powder obtained (75 µm) was made into nanoparticles using mechano-chemical, top-down method of nanoparticles synthesis with the presence of the surfactant BS-12 (dodecyl dimethyl bataine). The phase purity and crystallographic structures, the chemical functionality and the thermal characterization of the scaffolds’ powder were recognized using X-Ray Diffractometer (XRD), Fourier transform infrared (FTIR) spectrophotometer and Differential Scanning Calorimetry (DSC) respectively. Characterizations of the scaffolds were assessed by Scanning Electron Microscopy (SEM), Degradation Manner, Water Absorption Test, Swelling Test, Mechanical Test and Porosity Test. Top-down method produced cockle shell nanoparticles having averagely range 37.8±3–55.2±9 nm in size, which were determined using Transmission Electron Microscope (TEM). A mainly aragonite form of calcium carbonate was identified in both XRD and FTIR for all scaffolds, while the melting (Tm) and transition (Tg) temperatures were identified using DSC with the range of Tm 62.4–75.5 °C and of Tg 230.6–232.5 °C. The newly prepared scaffolds were with the following characteristics: (i) good biocompatibility and biodegradability, (ii) appropriate surface chemistry and (iii) highly porous, with interconnected pore network. Engineering analyses showed that scaffold 5211 possessed 3D interconnected homogenous porous structure with a porosity of about 49%, pore sizes ranging from 8.97 to 337 µm, mechanical strength 20.3 MPa, Young's Modulus 271±63 MPa and enzymatic degradation rate 22.7 within 14 days.     

Characterization and evaluation of hydrophobically modified chitosan scaffolds: Towards design of enzyme immobilized flow-through electrodes

Chitosan scaffolds were fabricated by application of thermally induced phase separation from aqueous solutions of unmodified chitosan and hydrophobically modified chitosan polymer. The final pore structure, in terms of diameter and geometry, were correlated to freezing temperature and freezing time for both the unmodified and hydrophobically modified chitosan polymer. Results showed that the resulting pore structure is strongly dependent upon the freezing temperature and less dependant upon the freezing time. For scaffolds produced from unmodified chitosan, the pore size decreased as expected with decreasing freezing temperature from ?5 °C to ?10 °C. However, an inconsistency in this trend was observed as the freezing temperature was decreased to ?20 °C. Combined analysis of pore size distribution and average pore diameter suggested that the freezing process was mainly mass transfer dominated at ?5 °C and ?10 °C, but principally heat transfer dominated at ?20 °C. In comparison, the scaffolds produced from hydrophobically modified chitosan (butyl-chitosan) followed the expected trend of decreasing mean pore diameter with decreased freezing temperatures throughout the entire temperature range. The scaffolds produced from the unmodified chitosan were more stable and rigid, and possessed average pore diameters that were generally smaller than those fabricated from the hydrophobically modified chitosan. The generally larger pores in the butyl-modified chitosan scaffolds might be explained by increased phase separation rates due to the introduced hydrophobicity of the chitosan polymer. Among the scaffolds fabricated from the butyl-modified chitosan, those produced at ?20 °C yielded the most uniform pore structure, the smallest average pore diameters, and the least temporal broadening of pore size distribution.     

Increase in rate of force development with skin cooling during isometric knee extension

Rate of force development (RFD) plays an important role when performing rapid and forceful movements. Cold-induced afferent input with transient skin cooling (SC) can modulate neural drive. However, the relationship between RFD and SC is unknown. The purpose of this study was to investigate whether SC increases RFD during isometric knee extension. Fifteen young healthy men (25 ± 8 yrs old) contracted their quadriceps muscle as fast and forcefully as possible with or without SC. Skin cooling was administered to the front of the thigh. Torque and electromyographic activity were measured simultaneously. Peak torque was not affected by SC. Skin cooling induced a significant increase in RFD at the phase 0–30 and 0–50 ms. The root mean square of the electromyography of vastus medialis, rectus femoris and vastus lateralis at the phases 0–30–50–100 ms increased significantly or tended to increase with SC. These results suggest that SC may increase neural drive and improve RFD in the very early phases of contraction.     

Effect of cooling rate and equilibration time on pre-freeze and post-thaw survival of buck sperm

Survival of buck sperm is affected due to duration and temperature of stages of refrigerated or frozen storage. This study investigated interactive effect of cooling rates (moderate; MC and rapid cooling; RC); and equilibration times (0, 2, 4 and 8 h) on survival before freezing at 4 °C and post-thaw quality of buck sperm. Semen was collected (three Beetal bucks; replicates = 6), pooled and diluted with Tris-citrate extender. Pooled semen samples were subjected to either RC (−2.2 °C/min) or MC (−0.3 °C/min) from 37 °C to 4 °C in separate aliquots and further equilibrated at 4 °C for 8 h. Semen was frozen using standard procedure after completion of each equilibration period i.e. 0, 2, 4 and 8 h. Semen was evaluated for motility, viability, plasma membrane integrity (PMI) and normal apical ridge (NAR) before freezing and after thawing. The survival time (time for survival above threshold limit i.e. 60%) at 4 °C, of motility and PMI was observed 5 and 6 h respectively in RC group while >8 h in MC group. Rate of decline (slope) in motility and viability was higher (P < 0.05) in RC overtime during equilibration at 4 °C while PMI and NAR declined at equal rate in both cooling groups. Post-thaw motility and NAR were higher (P < 0.05) in MC when equilibrated for 2–8 h while viability and PMI of RC was observed equal to MC group. In conclusion, survival of buck sperm is higher when cooled with moderate rate. However, RC can maintain post-thaw sperm viability and PMI equal to MC when equilibrated for 2–8 h. The methods should be explored to maintain motility and NAR during rapid cooling of buck sperm.     

Responses in acral and non-acral skin vasomotion and temperature during lowering of ambient temperature

Arteriovenous anastomoses (AVA) in acral skin (palms and soles) have a huge capacity to shunt blood directly from the arteries to the superficial venous plexus of the extremities. We hypothesized that acral skin, which supplies blood to the superficial venous plexus, has a stronger influence on blood flow adjustments during cooling in thermoneutral subjects than does non-acral skin. Thirteen healthy subjects were exposed to stepwise cooling from 32 °C to 25 °C and 17 °C in a climate chamber. Laser Doppler flux and skin temperature were measured simultaneously from the left and right third finger pulp and bilateral upper arm skin. Coherence and correlation analyses were performed of short-term fluctuations at each temperature interval. The flux from finger pulps showed the synchronous spontaneous fluctuations characteristic of skin areas containing AVAs. Fluctuation frequency, amplitude and synchronicity were all higher at 25 °C than at 32 °C and 17 °C (p<0.02). Bilateral flux from the upper arm skin showed an irregular, asynchronous vasomotor pattern with small amplitudes which were independent of ambient temperature. At 32 °C, ipsilateral median flux values from the right arm (95% confidence intervals) were 492 arbitrary units (au) (417, 537) in finger pulp and 43 au (35, 60) in upper arm skin. Flux values gradually decreased in finger pulp to 246 au (109, 363) at 25 °C, before an abrupt fall occurred at a median room temperature of 24 °C, resulting in a flux value of 79 au (31, 116) at 17 °C. In the upper arm skin a gradual fall throughout the cooling period to 21 au (13, 27) at 17 °C was observed. The fact that the response of blood flow to ambient cooling is stronger in acral skin than in non-acral skin suggests that AVAs have a greater capacity to adjust blood flow in thermoneutral zone than arterioles in non-acral skin.     

Operation of a fixed-bed bioreactor in batch and fed-batch modes for production of inulinase by solid-state fermentation

This work is focused on the inulinase production by solid-state fermentation (SSF) in a fixed-bed reactor (34 cm diameter and 50 cm height) with working capacity of 2-kg of dry substrate operated in batch and fed-batch modes. It was investigated different strategies for feeding the inlet air in the bioreactor (saturated and unsaturated air) as alternative to remove the metabolic heat generated during the microbial growth by evaporative cooling. The kinetic evaluation of the process carried out in batch mode using unsaturated air showed that the evaporative cooling decreasing the mean temperature of the solid-bed, although the enzyme production was lower than that obtained using saturated air. Results showed that maximum enzyme activity (586 ± 63 U gds⁻¹) was obtained in the fed-batch mode using saturated air after 24 h of fermentation. The enzymatic extract obtained by fed-batch mode was characterized and presented optimum temperature and pH in the range of 52–57 °C and 4.8–5.2, respectively. For a temperature range from 40 to 70 °C the enzyme presented decimal reduction time, D-value, ranging from 5748 to 47 h, respectively. For a pH range from 3.5 to 5.5 the enzyme showed good stability, presenting D-values higher than 2622 h. In terms of Michaelis–Mentem parameters were demonstrated that the crude inulinase activity presented higher affinity for substrate sucrose compared to inulin.     

Hatching,survival and deformities of piracanjuba (Brycon orbignyanus) embryos subjected to different cooling protocols

Groups of one hundred Brycon orbignyanus embryos at the stage of blastopore closure were subjected to different cooling protocols. Different combinations and concentrations of cryoprotectants were tested: sucrose, methanol, ethylene glycol and dimethyl sulfoxide (Me₂SO); at different temperatures (0.0 ± 2.0 °C and 8.0 ± 2.0 °C) and refrigeration times (6, 10, 24, 72 and 168 h), with the exception of the positive control (incubation without previous cooling). At the end of each refrigeration time, the embryos were acclimatized, rehydrated and incubated to determine hatching, survival and deformity rates. Morphological analysis of embryos was also carried out. The results showed that temperature and refrigeration time are critical factors for embryo survival. No embryos survived after 24, 72 and 168 h of refrigeration. Furthermore, when the refrigeration time increased from 6 to 10 h and the temperature decreased from 8.0 ± 2.0 °C to 0.0 ± 2.0 °C, mortality rates increased significantly. It was also found that in all protocols dead eggs and/or larvae with some degree of deformity were present. The main larval deformities observed were the malformation of the head, tail, yolk sac, vertebral column and eyes.     

Synthesis and sorption performance of highly specific imprinted particles for the direct recovery of carminic acid

Carminic acid (CA) is a colorant of natural origin, which is demanded by the food, cosmetic, and pharmaceutical industries. In this work, a selective finite bath process was developed based on the utilization of molecularly imprinted polymeric particles. Such adsorbent was synthesized in a (porous) particle shaped form employing methacrylic acid (MAA) and 4-vinylpyridine (4Vpy) as monomers and ethylene glycol dimethacrylate (EDGMA) as a cross-linker. The imprinted particles were characterized by surface area, surface charge and pore size determination. The adsorption behavior of CA on such a material followed a Langmuir–Freundlich isotherm. Maximum capacity at equilibrium reached 0.64 mmol/g (316 mg/g) and maximum available binding sites 1.8 mmol/g (917 mg/g) were observed for an association coefficient value of 1.5 mM^?1. Further, the imprinting factor; showing the strength of interaction of CA to the polymer, was calculated as 13 while the selectivity factor depicted a value of 15. The data presented indicates that the imprinted adsorbent could be conveniently utilized for the recovery of CA, from cochineal extract, in the finite bath mode of operation.     

Variability of sea-surface temperature and sea-ice cover in the Fram Strait over the last two millennia

A sediment core located on the West Spitzbergen margin in the Fram Strait (78°54.931′N, 6°46.005′E, water depth: 1497 m) was analyzed for its dinocyst content in order to reconstruct hydroclimatic variations of the last 2500 years. The relative abundance of dinocyst taxa and principal component analysis show a major transition at about 300 cal. years BP. It is characterized by the disappearance of thermophilic taxa Spiniferites mirabilis-hyperacanthus and Impagidinium sphaericum and the increase of polar–subpolar taxa Impagidinium pallidum and Pentapharsodinium dalei. Sea-surface temperature (SST) estimates suggest warmer conditions than present (anomaly～+2 °C) averaging at 7 °C in summer until 300 cal. years BP, although cooling pulses are recorded around 1700, 1500, 1200 and 800 cal. years BP. The last 300 years were marked by a cooling from 7.6 to 3.5 °C and sea-ice cover increasing up to 7 months/yr. The results demonstrate that the Fram Strait area is sensitive to hydroclimatic variations, notably with respect to sea-ice and SSTs, which are linked to the relative strength of northward flow of North Atlantic waters to the East and southward outflow of cold and fresh waters from the Arctic Ocean. Based on our data, the warmest part of our record around 1320 cal. years BP is the only interval of the last 2500 years that provides a possible analogue for the modern post-AD 2000 interval, which is characterized by sea-ice free conditions.     

Injury due to extravasation of thiopental and propofol: Risks/effects of local cooling/warming in rats

Inadvertent leakage of medications with vesicant properties can cause severe necrosis in tissue, which can have devastating long-term consequences. The aim of this study was to evaluate the extent of extravasation injury induced by thiopental and propofol, and the effects of cooling or warming of local tissue on extravasation injury at macroscopic and histopathologic levels. Rats were administered intradermally thiopental (2.5 mg/100 µL) or propofol (1.0 mg/100 µL). Rats were assigned randomly to three groups: control (no treatment), cooling and warming. Local cooling (18–20 °C) or warming (40–42 °C) was applied for 3 h immediately after agent injection. Lesion sizes (erythema, induration, ulceration, necrosis) were monitored after agent injection. Histopathology was evaluated in skin biopsies taken 24 h after agent injection. Thiopental injection induced severe skin injury with necrosis. Peak lesions developed within 24 h and healed gradually 18–27 days after extravasation. Propofol induced inflammation but no ulceration, and lesions healed within 1–2 days. Local cooling reduced thiopental- and propofol-induced extravasation injuries but warming strongly exacerbated the skin lesions (e.g., degeneration, necrosis) induced by extravasation of thiopental and propofol. Thiopental can be classified as a “vesicant” that causes tissue necrosis and propofol can be classified as an “irritant”. Local cooling protects (at least in part) against skin disorders induced by thiopental and propofol, whereas warming is harmful.     

Towards whole sheep ovary cryopreservation

Cryopreservation of ovarian tissue aims to assist young women who require treatments that may lead to sterility or infertility. Cryopreservation procedures should therefore be as simple and efficient as possible. This study investigates rapid cooling outcomes for whole sheep ovaries. Ovaries were perfused with VS4 via the ovarian artery, and cooled by quenching in liquid nitrogen in less than a minute (estimated cooling rate above 300 °C/min till the vitreous transition temperature). The ovaries were rewarmed in two stages: slow warming (12–16 °C/min from −196 to −133 °C) in liquid nitrogen vapour, followed by rapid thawing in a 45 °C water bath at about 200 °C/min. DSC measurements showed that under these cryopreservation conditions VS4 would vitrify, but that VS4 perfused ovarian cortex fragments did not vitrify, but formed ice (around 18.4%). Immediately following rewarming, a dye exclusion test indicated that 61.4 ± 2.2% of small follicles were viable while histological analysis showed that 48 ± 3.8% of the primordial follicles were normal. It remains to be clarified whether follicle survival rates will increase if conditions allowing complete tissue vitrification were used.     

Stability and structural changes of horseradish peroxidase: Microwave versus conventional heating treatment

Effects of conventional heating (CH) and microwave (MW) on the structure and activity of horseradish peroxidase (HRP) in buffer solution were studied. CH incubation between 30 and 45 °C increased activity of HRP, reaching 170% of residual activity (RA) after 4–6 h at 45 °C. CH treatment at 50 and 60 °C caused HRP inactivation: RA was 5.7 and 16.7% after 12 h, respectively. Secondary and tertiary HRP structural changes were analyzed by circular dichroism (CD) and intrinsic fluorescence emission, respectively. Under CH, activation of the enzyme was attributed to conformational changes in secondary and tertiary structures. MW treatment had significant effects on the residual activity of HRP. MW treatment at 45 °C/30 W followed by CH treatment 45 °C regenerated the enzyme activity. The greatest loss in activity occurred at 60 °C/60 W/30 min (RA 16.9%); without recovery of the original activity. The inactivation of MW-treated HRP was related to the loss of tertiary structure, indicating changes around the tryptophan environment.     

Gelation of high methoxy pectin by acidification with d-glucono-δ-lactone (GDL) at room temperature

Rheological comparisons have been made between preparations of high methoxy pectin (DE ≈ 70%) gelled by acidification with d-glucono-δ-lactone (GDL) on holding for 16 h at 25 °C in the presence of 60 wt% sucrose, and otherwise identical preparations gelled by acidification with citric acid at high temperature and cooling from 90 to 25 °C at 1 °C/min. Two series of experiments were carried out for both methods of acidification. In the first series, the concentration of pectin (c) was held constant at 1.0 wt% and the final pH attained after holding (with GDL) or cooling (with citric acid) was varied from 3.75 to 2.25. In the second series, the final pH was held constant at 3.0 and c was varied from 0.25 to 2.00 wt%. All samples were then heated (1 °C/min) from 25 to 90 °C. Rheological changes on cooling/holding and heating were characterised by low-amplitude oscillatory measurements of storage modulus (G′) and loss modulus (G′′) at 1 rad s^?1 and 0.5% strain, and mechanical spectra were recorded at 25 °C. Selected samples, gelled with GDL, were also characterised by compression testing (at 25 °C), and a direct linear relationship was found between the logarithm of yield stress and log G′.The concentration-dependence of moduli for the samples acidified to pH 3.0 with GDL had the form typical of biopolymer gels, with log G′ versus log c approaching a limiting slope of 2 as c was raised above the minimum critical gelling concentration (c_o ≈ 0.3 wt%). Under all conditions of pH and pectin concentration studied, the values of G′′ (at 25 °C) for the samples acidified with citric acid were higher than those of the corresponding GDL-induced networks. The values of G′ were also higher, except at very low pH (below ～2.7 at c = 1.0 wt%) or very high concentrations of pectin. At pectin concentrations above ～1.5 wt%, the moduli of the samples gelled with citric acid (at pH 3.0) levelled out, or decreased slightly, with the values of G′ dropping below those of the GDL-induced networks towards the end of the concentration range studied (at c ≈ 2 wt%). All samples acidified with citric acid showed gel-like response (G′ > G′′) at 90 °C, attributed to hydrophobic association. The downturn in moduli at 25 °C for high concentrations of pectin is attributed to formation and disruption of strong networks during mixing with citric acid at high temperature (“pregelation”). It is suggested, however, that “weak gels” formed at lower concentrations or at pH values above ～2.7 may enhance gel properties by preserving a continuous network as hydrophobic junctions dissociate on cooling and are replaced by hydrogen-bonded junctions, in contrast to random percolation during gelation with GDL at 25 °C. On re-heating from 25 to 90 °C, the reverse processes (dissociation of hydrogen-bonded structures and formation of hydrophobic associations) were evident in an initial reduction and subsequent increase in moduli, as observed in previous studies. Similar heating traces were obtained for samples acidified with GDL to pH values above ～3.0 (at c = 1.0 wt%) or with pectin concentrations below ～1.0 wt% (at pH 3.0). However, at higher concentrations or lower values of pH (i.e. conditions favourable to extensive intermolecular association) an abrupt decrease in G′, with an accompanying maximum in G′′, was observed on heating through the temperature range ～60–80 °C. This is attributed to excessive hydrophobic association, causing collapse of network structure. It is further suggested that, for samples acidified with citric acid, there is preferential association of chain sequences of high ester content into hydrophobic junctions at 90 °C, leaving sequences with a high content of unesterified carboxyl groups available to form long hydrogen-bonded junctions during cooling, and thus giving gels that are stronger and more resistant to network collapse.     

Growth performance and carcass traits of forage-fed hair sheep wethers

The objective of the present study was to compare live animal performance and carcass characteristics of 3/4 or 7/8 Dorper (DO; n = 30), purebred Katahdin (KA; n = 20), purebred St. Croix (SC; n = 17) and purebred Suffolk (SU; n = 10) lambs born in the spring and fall of 2001. After weaning, lambs were supplemented with up to 680 g of a corn-soybean meal supplement while grazing bermudagrass pastures overseeded with ryegrass. Lambs were slaughtered at approximately 210 d of age. From birth to weaning, DO lambs gained faster (P < 0.001) than KA or SC lambs, whereas KA lambs had higher (P < 0.001) ADG than SC lambs. Additionally, DO and SU wethers had greater (P < 0.02) ADG from weaning to slaughter than SC or KA wethers. Suffolk lambs were heavier (P < 0.001) at slaughter and produced heavier (P < 0.001) carcasses than lambs from hair-sheep breeds. Carcasses of KA lambs were fatter (actual fat thickness; P < 0.02) resulting in higher yield grades (P < 0.03) than carcasses of DO, SC, or SU lambs. Carcasses of DO and SU had larger (P < 0.001) longissimus muscle (LM) areas than those of KA or SC carcasses, whereas kidney fat weight and percentage were greater (P < 0.001) in carcasses from KA and SC than DO and SU lambs. Lean maturity was similar (P = 0.32) among breed-types. However, skeletal maturity was greater (P < 0.001) in SU than hair-sheep carcasses. Flank-streaking scores were similar (P = 0.19) among the breed-types, but conformation scores were higher (P < 0.001) for DO and SU carcasses and resulted in higher (P < 0.001) quality grades than SC carcasses. The LM of SU lambs was lighter (higher L^* values; P < 0.05) than that of KA and SC lambs, whereas the LM from DO lambs was redder (higher a^* values; P < 0.001) than SC and SU and more (P < 0.001) yellow than that of the other breed-types. Chops from SU lambs were tougher (higher shear force values; P < 0.007) than chops from the hair-sheep breeds. Results of this study indicate that ADG, carcass muscularity and meat quality were similar between DO and SU lambs, and, although fatter, carcass muscularity of KA was similar to that of DO lambs.     

Braking characteristics during cutting and pivoting in female soccer players

Most biomechanical studies into changing direction focus on final contact (FC), whilst limited research has examined penultimate contact (PEN). The aim of this study was to explore the kinematic and kinetic differences between PEN and FC of cutting and pivoting in 22 female soccer players (mean ± SD; age: 21 ± 3.1 years, height: 1.68 ± 0.07 m, mass: 58.9 ± 7.3 kg). Furthermore, the study investigated whether horizontal force–time characteristics during PEN were related to peak knee abduction moments during FC. Three dimensional motion analyses of cutting and pivoting on the right leg were performed using Qualysis ‘Proreflex’ infrared cameras (240 Hz). Ground reaction forces (GRF) were collected from two AMTI force platforms (1200 Hz) to examine PEN and FC. Both manoeuvres involved significantly (P < 0.05) greater knee joint flexion angles, peak horizontal GRF, but lower average horizontal GRF during PEN compared to FC. Average horizontal GRF during PEN (R = −0.569, R² = 32%, P = 0.006) and average horizontal GRF ratio (R = 0.466, R² = 22%, P = 0.029) were significantly related to peak knee abduction moments during the FC of cutting and pivoting, respectively. The results indicate PEN during pre-planned changing direction helps reduce loading on the turning leg where there is greater risk of injuries to knee ligaments.     

Hybrid cooling vest for cooling between exercise bouts in the heat: Effects and practical considerations

While continuous cooling strategies may induce some ergonomic problems to occupational workers, cooling between work bouts may be an alternative for cooling them down in hot environments. The purpose of this study was to assess the effects of wearing a newly designed hybrid cooling vest (HCV) between two bouts of exercise. Inside a climatic chamber set at an air temperature of 37 °C and a relative humidity of 60%, twelve male participants underwent two bouts of intermittent exercise interspersed with a 30 min between-bout recovery session, during which HCV or a passive rest without any cooling (PAS) was administered. The results indicated that thermoregulatory, physiological, and perceptual strains were significantly lower in HCV than those in PAS during the recovery session (p≤0.022), which were accompanied with a large effect of cooling (Cohen's d=0.84–2.11). For the second exercise bout, the exercise time following HCV (22.13±12.27 min) was significantly longer than that following PAS (11.04±3.40 min, p=0.005, d=1.23) During this period, core temperature T_c was significantly lower by 0.14±0.0.15 °C in HCV than that in PAS. The heart rate drift over time was declined by 2±2 bpm min⁻¹ (p=0.001, d=1.00) and the rise in physiological strain index was reduced by 0.11±0.12 unit min⁻¹ (p=0.010, d=0.96) following the use of HCV. These findings suggested that using HCV could accelerate between-bout recovery and improve subsequent exercise performance by the enlarged body core temperature margin and blunted cardiovascular drift.     

Characterization and antioxidant activity for exopolysaccharide from submerged culture of Boletus aereus

Three polysaccharide fractions (designated as Fr-I, Fr-II and Fr-III) were successfully purified from the crude exopolysaccharide (EPS) produced from submerged culture of Boletus aereus by gel filtration chromatography on Sepharose CL-6B. The size exclusion chromatography/multi-angle laser light scattering (SEC/MALLS) system showed that the average molecular weights (Mws) of these three fractions were 1.365 × 10⁶, 1.048 × 10⁵ and 2.471 × 10⁴ g/mol, respectively. The SEC/MALLS also revealed that the molecular conformation of the Fr-I was a random coil, with Fr-II being a rigid rod in aqueous solution. Moreover, monosaccharide composition analysis indicated that Fr-I was mainly composed of glucose, while both of Fr-II and Fr-III were mainly composed of mannose and glucose. Then, FT-IR spectral analysis of the purified EPS revealed prominent characteristic groups. Furthermore, thermo gravimetric analysis (TGA) indicated the degradation temperature of the Fr-I (170 °C) was higher than those of Fr-II (156 °C) and Fr-III (155 °C). Finally, on the basis of the antioxidant activity test in vitro, Fr-I exhibited the highest antioxidant ability among these samples, which might be attributed to the monosaccharide composition and molecular weight in the EPS fraction.     

Biological control of Botrytis bunch rot in Atlantic climate vineyards with Candida sake CPA-1 and its survival under limiting conditions of temperature and humidity

Candida sake CPA-1 is an antagonistic yeast that has previously been shown to effectively control Botrytis bunch rot in grapes. The efficacy of biological control agents is dependent on their survival, which may also depend on climatic conditions. However, few studies have evaluated the effect of abiotic factors affecting the survival of biological control agents, such as temperature (T) or relative humidity (RH). In this study, efficacy of C. sake (5 × 10⁷ CFU mL⁻¹), which was applied with the additive Fungicover (FC; 50 g L⁻¹), was tested against BBR in the laboratory and in field trials under the Atlantic climate conditions of the Bordeaux region (France). The study also evaluated the survival of C. sake under T and RH regimes simulated in climatic chambers. Two or five applications of C. sake plus FC during the growing season significantly reduced BBR severity at harvest by 48% and 82%, respectively, when compared to the control. Similar reductions were achieved after inoculation with selected virulent Botrytis cinerea strains (75% compared to control) in laboratory experiments. C. sake populations showed minimal decreases between field applications and were favored by simulated Atlantic climate conditions. The survival pattern of C. sake exposed to 40 and 45 °C combined with 30% and 100% of RH was described, demonstrating a sharp decrease during the first 24 h. Allowing 48 h for C. sake to incubate and become established on fruits prior to the exposure to 40 °C and 30% RH increased survival (P < 0.05). These results confirm the efficacy of treatment with C. sake plus FC under favorable climatic conditions for BBR development, while survival studies may help to improve the survival and efficacy of yeast BCAs, such as C. sake CPA-1.     

Cryopreservation of sperm from seven-band grouper,Epinephelus septemfasciatus

In the present study, we examined methods for the cryopreservation of Epinephelus septemfasciatus spermatozoa. The percent motility, average path velocity, and linearity of movement (LIN) of fresh and corresponding post-thaw sperm were evaluated. Sperm motility was investigated using computer-assisted sperm analysis. Five percent dimethyl sulphoxide (Me₂SO) with 95% fetal bovine serum (FBS) was the most successful cryoprotectant diluent with a comparative post-thaw motility of 77.6 ± 8.5%; 5% dimethyl formamide was also effective. Fetal bovine serum was significantly better as an extender when compared with artificial seminal plasma, glucose, and trehalose solution. Sperm tolerated a wide range of cooling rates (from 27.1 to 94.3 °C min^?1); however, the post-thaw motility of sperm cooled to ?30 °C was significantly lower than that of other cooled temperatures (?40 to ?70 °C). The velocity of post-thaw sperm was significantly lower than that of fresh sperm, although LIN remained the same. For effective cryopreservation of seven-band grouper sperm, samples should be diluted in 5% Me₂SO with 95% FBS and cooled to at least ?40 °C before immersion in liquid nitrogen.     

Serum concentrations of free amino acids in growing pigs exposed to diurnal heat stress fluctuations

In areas where the ambient temperature (AT) is above the thermo neutral (TN) zone of pigs, significant changes within a 24-h period occur, differently affecting the availability of amino acids (AA) within the same day. An experiment was conducted to analyze the serum concentrations (SC) of free AA in pigs exposed to diurnal variations in AT. Six pigs (27.1 ±1.3 kg body weight) implanted with a thermometer to register the body temperature (BT) at 15-min intervals were used. Blood samples were collected on the last 3 d of the 14-d study, at 0700 h (lowest AT), 1200 h (mild HS), and 1600 h (severe HS). The pigs received 1.2 kg/d of an AA-supplemented, wheat-soybean meal diet, in two equal meals (0700 and 1900 h). The AT and BT, recorded at 0700, 1200, and 1600 h was: 30.6, 38.6, 41.1 °C, and 38.2, 39.5, 40.3 °C, respectively. The BT was significantly correlated (P < 0.001) with the AT. The SC (μM/mL) of Ile, Lys, Met, Val, Ala, Asn, and Pro were higher (P ≤ 0.01); Arg, Phe, Glu, and Tyr tended to be higher (P ≤ 0.10); but Cys was lower (P < 0.05) at 1200 h than at 0700 h. Lys was higher, Cys and Tyr were lower (P < 0.05), and Ile and Val tended to be higher (P ≤ 0.10) at 1600 h than at 0700 h. Serum Arg, Ile, Phe, Ala, Asn, Gln, Pro, Ser, and Tyr were lower (P < 0.05), and Leu and Val tended to be lower at 1600 h than at 1200 h. These data demonstrate that AT directly alters the BT of pigs, and that diurnal variations in AT differently affect their SC and availability of AA for growth.