Oxygen enhances the recovery of Potamogeton maackianus from prolonged exposure to very low irradiance

Recovery ability in relation to carbohydrate content of Potamogeton maackianus growing in two dissolved oxygen concentrations (8 and 2 mg L⁻¹) was investigated during 28 days exposure to very low irradiance (about 0.06 μmol m⁻² s⁻¹). Plant weight remained relatively constant (0.19 g dry wt plant⁻¹) within the initial 21 days in the high oxygen treatment, but decreased to 0.14 g dry wt plant⁻¹ at the end of the experiment. In low oxygen environments, plant weight was similar within the initial 14 days, but decreased to 0.08 g dry wt plant⁻¹ at 21 day. During the experimental period, both soluble sugar and starch contents in shoots decreased with time. Compared to high oxygen treatment, plants in the low oxygen treatment depleted starch more quickly (25 versus 18 mg g⁻¹ at 28 day) but remained a relatively high soluble sugar content (0.9 versus 1.8 mg g⁻¹ at 28 day). After recovery in high light and high dissolved oxygen conditions for 1 week, plant growth rate, new branch number, stem elongation rate and leaf recruitment number were significantly higher in high oxygen than in the low oxygen treatments. These data suggest that the ability of the plant to recover from prolonged exposure to very low irradiance is related to the depletion level of carbohydrate stored in plant tissues, which is regulated by oxygen availability in the water.     

Coupled effects of irradiance and iron on the growth of a harmful algal bloom-causing microalga Scrippsiella trochoidea

Effects of irradiance and iron on the growth of a typical harmful algal blooms (HABs) causative dinoflagellate, Scrippsiella trochoidea, were investigated under various irradiances (high light: 70 μmol m^?2 s^?1 and low light: 4 μmol m^?2 s^?1) and iron concentrations (low iron: 0.063 mg L^?1, medium iron: 0.63 mg L^?1 and high iron: 6.3 mg L^?1), and evaluated by the parameters of algal cell density, specific growth rate, optical density and chlorophyll a content. The results indicated that there was significant difference in the cell density of dinoflagellate S. trochoidea between high light and low light intensity treatments across the entire experiments, 7-fold higher at high irradiance as compared with low irradiance, which was further enhanced by the iron concentration. It was found that the maximum cell density of 25 × 10⁴ cell mL^?1 occurred under the combination of high light intensity and high iron concentration, followed by 23 × 10⁴ cell mL^?1 in the combination of high light and medium iron, and 20 × 10⁴ cell mL^?1 in the combination of high light and low iron. There was no significant effect of iron concentration on the cell density under low light intensity. The cell density maintained about 3 × 10⁴ cell mL^?1 across all combinations of iron concentrations and low light in the end of experiments. Such interactive effects of light intensity and iron level dependent were also observed for the specific growth rate, OD₆₈₀ and chlorophyll a content of S. trochoidea. The maximum values of specific growth rate, OD₆₈₀ and chlorophyll a content peaked at the condition of high irradiance and high iron, which were 0.22 d^?1, 0.282 and 0.673 mg L^?1, respectively. In general, their values increased significantly with the increasing of iron concentration at high irradiance, whereas no significant difference was observed among three iron concentrations at low irradiance, all remaining approximately 0.06 d^?1, 0.03 and 0.050 mg L^?1, respectively. Those results suggest that there may be a strong interactive effect between irradiance and iron on microalgal growth and their physiological characteristics. The combination of high light and high iron concentration may accelerate algal cell growth and pigment biosynthesis, thus leading to massive occurrence of HABs.     

Wastewater effects on Phaeodactylum tricornutum (Bohlin): Setting up a classification system

Phytoplankton is highly productive in marine coastal ecosystems that generally present high levels of anthropogenic pressures. Microalgae presents very high surface-to-volume ratio and may respond promptly to contaminants showing either an increase in growth or inhibition effects. Thus phytoplankton organisms provide information on the potential impacts of contaminants on the supported marine-coastal food webs. The diatom Phaeodactylum tricornutum Bohlin is commonly used in toxicity testing according to the ISO 10253:2006 standardised protocol, but a comprehensive inventory of tested substances has not been compiled yet. The aim of this study is to establish a wastewater effect score based on P. tricornutum exposed to domestic, municipal and industrial wastewaters from activated sludge sequencing batch reactor and ultra-filtration membrane biological reactors. Wastewater samples produced stimulation and inhibition effects identified by biostimulation unit (BU₅₀) and toxicity unit (TU₅₀) both at 50% effect, respectively. Within the stimulation scenario, toxicity was low if 0 < BU₅₀ ≤ 0.31, medium if 0.31 < BU₅₀ ≤ 1.05, high if 1.05 < BU₅₀ ≤ 1.64, and very high if BU₅₀ > 1.64. Within the inhibition scenario, toxicity was low if 0 < TU₅₀ ≤ 0.07, medium if 0.07 < TU₅₀ ≤ 2.67, high if 2.67 < TU₅₀ ≤ 5.86, and very high if TU₅₀ > 5.86. Results evidenced that nitrogen and phosphorus concentrations were not correlated to ecotoxicological values, probably due to the presence of undetected micronutrients, confirming the importance of toxicity-based hazard assessment.     

Cognition in female rats after blocking conversion of androgens to estrogens

Women and non-human females have surprisingly high levels of circulating testosterone, yet the effects of androgens on non-reproductive behaviors, including cognition, of females are not well characterized. The current project used an aromatase inhibitor, letrozole, to block conversion of androgens to estrogens. Adult female rats were ovariectomized and administered either vehicle only, testosterone propionate only (400 μg/kg, TP only), letrozole only (1 mg/kg, Letro only), or the combination of letrozole and testosterone (TP + Letro) over 4 weeks. A gonadally intact group was used for comparisons. During the last 3 weeks, the animals were tested for working memory in both a spatial task (radial arm maze) and a non-spatial task (object recognition). At sacrifice, uterine weights and serum testosterone and estradiol were determined. Behavioral results were the intact animals showed better working memories on the object recognition task, but that there were no differences among the ovariectomized groups. In the radial arm maze task, groups with best to worst performance were TP only > Intact = TP + Letro > vehicle = Letro only. Highest to lowest serum titers, for testosterone, were TP + Letro > TP only > Intact = Letro only > vehicle and, for estradiol, Intact > TP only > Vehicle > Letro only = TP + Letro. Our interpretation is that testosterone enhanced spatial performance when bioavailability of both TP and E2 are high, and high testosterone can rescue spatial memory when E2 bioavailability is low.     

Tolerance of hairy roots of carrots to U chronic exposure in a standardized in vitro device

In soil, high variability of U bioavailability results in large range of apparent U toxic levels for plants. U toxicity on hairy roots of carrot was studied in nutrient gel with a standardized in vitro device. After exposure to 2.5 and 20 mg U L^?1 for 34 days, U concentration ranged between 4 and 563 mg U kg^?1 fresh weight which was in good accordance with U accumulation by roots of plant from contaminated soils. Threshold of U toxicity for root length decreased with time and a transient hormesis occurred for exposure to 2.5 and 5 mg U L^?1. After 34 days and with root length as endpoint, significant toxicity appeared at a gel contamination level above 7.5 mg U L^?1 corresponding to a maximum U concentration in the liquid phase of 0.8 mg L^?1. The calculated EC50 for root length as a function of gel contamination was 9.4 mg U L^?1. Lower threshold and EC50 were observed for biomass as endpoint (resp. 5 and 7.3 mg U L^?1). The low values observed in this study could result from high sensitivity of carrot to U, high bioavailability of U in gel or absence of interferences with microorganisms. This in vitro device appeared adapted to study toxicity of U to plant roots in optimal conditions of both exposure and observations and is recommended to examine further physiological processes and the influence of microorganism interactions.     

Effect of acetazolamide on cytokines in rats exposed to high altitude

Acute mountain sickness (AMS) is a dangerous hypoxic illness that can affect humans who rapidly reach a high altitude above 2500 m. In the study, we investigated the changes of cytokines induced by plateau, and the acetazolamide (ACZ) influenced the cytokines in rats exposed to high altitude. Wistar rats were divided into low altitude (Control), high altitude (HA), and high altitude + ACZ (22.33 mg/kg, Bid) (HA + ACZ) group. The rats were acute exposed to high altitude at 4300 m for 3 days. The HA + ACZ group were given ACZ by intragastric administration. The placebo was equal volume saline. The results showed that hypoxia caused the heart, liver and lung damage, compared with the control group. Supplementation with ACZ significantly alleviated hypoxia-caused damage to the main organs. Compared with the HA group, the biochemical and blood gas indicators of the HA + ACZ group showed no difference, while some cytokines have significantly changed, such as activin A, intercellular adhesion molecule-1 (ICAM-1, CD54), interleukin-1α,2 (IL-1α,2), l-selectin, monocyte chemotactic factor (MCP-1), CC chemokines (MIP-3α) and tissue inhibitor of matrix metalloproteinase 1 (TIMP-1). Then, the significant difference pro-inflammatory cytokines in protein array were chosen for further research. The protein and mRNA content of pro-inflammatory cytokines MCP-1, interleukin-1β (IL-1β), tumor necrosis factor (TNF-α), interferon-γ (IFN-γ) in rat lung were detected. The results demonstrated that the high altitude affected the body’s physiological and biochemical parameters, but, ACZ did not change those parameters of the hypoxia rats. This study found that ACZ could decrease the content of pro-inflammatory cytokines, such as MCP-1, IL-1β, TNF-α and IFN-γ in rat lungs, and, the lung injury in the HA + ACZ group reduced. The mechanism that ACZ protected hypoxia rats might be related to changes in cytokine content. The reducing of the pro-inflammatory cytokines in rat lung might be other reason to explain ACZ against the acute mountain sickness.     

Effects of irradiance and water flow on formation and growth of spongy and filamentous thalli of Codium fragile

Effects of irradiance and water flow on formation and growth of filamentous and spongy thalli of Codium fragile (Suringar) Hariot growing on vinylon threads were investigated at the laboratory culture. They showed clear differences in their irradiance and water flow requirements for their formation and growth. Spongy thalli were formed from the cultured filamentous thalli only at the high water flow velocity (10 cm s⁻¹). Number of the spongy thalli remarkably increased with increasing irradiance because those at 10, 50 and 100 μmol m⁻² s⁻¹ reached 0, 2 and 76 thalli m⁻¹, respectively, by 10 weeks of culture. In contrast, filamentous thalli were formed from the cultured spongy thalli at 0 and 3 cm s⁻¹, and difference in irradiance had no effect on their formation. Growth of the spongy thalli greatly accelerated under the combination of the high irradiance and high water velocity (200 μmol m⁻² s⁻¹ and 10 cm s⁻¹) because their relative growth rate in wet weight under the condition was two–four times higher than those at the other examined irradiances and water velocities. On the other hand, difference in water velocity had no effect on growth of the filamentous thalli under flowing water, and their growth decelerated at the high irradiance (200 μmol m⁻² s⁻¹). This demonstrates that water flow is a major factor controlling the formation of the spongy and filamentous thalli. The formation and growth of the spongy thalli surely occur under the combination of the high irradiance and fast flowing water. In contrast, the formation of the filamentous thalli occurs in the calm water, and their growth is inhibited under the high irradiance.     

Effect of high tannin grain sorghum on gastrointestinal parasite fecal egg counts in goats

The objective of three experiments was to determine the influence of high condensed tannin (CT) grain sorghum on gastrointestinal parasite fecal egg counts (FEC). Sixteen naturally infected Boer crossbred mixed-sex goats were used. Animals that were supplemented with grain daily were removed from pasture and placed in individual pens and fed treatment diets for 21 or 14 days (Experiment 3). Goats were allowed ad libitum access to water and diets containing high or low CT cracked grain sorghum. On day 0 and every 7 days thereafter, PCV, FEC and FAMACHA^® eyelid color scores (EYE; Experiment 1 only) were recorded. For Experiment 1, percentage of animals dewormed, PCV and EYE were not influenced by treatment and averaged 13.6 ± 4.5% per treatment period, 23.4 ± 0.8% and 3.2 ± 0.12, respectively, for all animals. The FEC increased after day 0 for control but not high tannin grain sorghum fed goats (treatment by day interaction, P < 0.02). As expected, EYE and PCV were negatively related (r = −0.45; P < 0.0005). For Experiment 2, there was no influence of diet on PCV or FEC, but there was an effect of day on FEC (P < 0.002) in which all goats had higher FEC on day 0 (1956 ± 219 eggs/g) than any other sampling day. For Experiment 3, there was no effect of feeding high CT grain sorghum on FEC (2992 ± 591 eggs/g) or PCV (25.1 ± 0.5%). In these studies, high CT grain sorghum did not consistently influence FEC and did not impact PCV or number of animals requiring chemical anthelmintic treatment.     

Microbial community in conventional and extended aeration activated sludge plants in India

This study was conducted to determine the relationship between the ciliated protozoan population density and the effluent quality at two different modes of activated sludge plants (ASP) operating in India. A wide variety of ciliated protozoa (26 sp.) in higher density were identified at the conventional ASP, Haridwar, that delivered high quality effluent in terms of low biochemical oxygen demand (BOD = 15 mg/L), suspended solids (SS = 17 mg/L), turbidity (2.7 NTU), ammoniacal nitrogen (NH₃–N = 3 mg/L), chemical oxygen demand (COD = 37 mg/L), total coliforms (TC = log 5.2), fecal coliforms (FC = log 4.7) and fecal Streptococci (FS = log 3.7). Whereas, a few protozoan species (15 sp.) in lower density were reported in extended aeration plant (EAP) Delhi, that delivered turbid and lower quality effluent in terms of high BOD (23 mg/L), SS (80 mg/L), turbidity (12 NTU), NH₃–N (55 mg/L) and COD (68 mg/L). However, in spite of relatively poor effluent quality, lower concentration of TC (log 4.2), FC (log 3.9) and FS (log 3.2) was observed in EAP, Delhi. The constant presence of two filamentous bacterial species (Beggiatoa and Spirillum) in extended aeration process can be considered as the probable reason of high coliforms removal, since filamentous bacteria are capable of removing organic as well as microbial pollutants from wastewater.     

Stability and activity of Dictyoglomus thermophilum GH11 xylanase and its disulphide mutant at high pressure and temperature

The functional properties of extremophilic Dictyoglomus thermophilum xylanase (XYNB) and the N-terminal disulphide-bridge mutant (XYNB-DS) were studied at high pressure and temperature. The enzymes were quite stable even at the pressure of 500 MPa at 80 °C. The half-life of inactivation in these conditions was over 30 h. The inactivation at 80 °C in atmospheric pressure was only 3-times slower. The increase of pressure up to 500 MPa at 80 °C decreased only slightly the enzyme's stability, whereas in 500 MPa the increase of temperature from 22 to 80 °C decreased significantly more the enzyme's stability. While the high temperature (80–100 °C) decreased the enzyme reaction with short xylooligosaccharides (xylotetraose and xylotriose), the high pressure (100–300 MPa) had an opposite effect. The temperature of 100 °C strongly increased the K_m but did not affect the k_cat to the same extent, thus indicating that the interaction of the substrate with the active site suffers before the catalytic reaction begins to decrease as the temperature rises. Circular dichroism spectroscopy showed the high structural stability of XYNB and XYNB-DS at 93 °C.     

Intra- and interday reliability of voluntary and electrically stimulated isometric contractions of the quadriceps femoris

The reliability of voluntary and electrically stimulated isometric contractions of m. quadriceps femoris of male participants (n = 10; age 30 ± 8 years; height 1.79 ± 0.05 m; body mass 79.4 ± 8.3 kg) was investigated using ratio limits of agreement (LoA) on a time scale common to examine recovery from muscle damaging exercise. No systematic changes in reliability occurred over time (baseline versus 2, 24, 48, and 72 h). Maximal voluntary contraction (MVC) and interpolated twitch technique (ITT) showed no mean bias (P > 0.05) with 95% LoA of ±12.7 and ±5.4, respectively. Resting twitch and potentiated doublet peak force showed no mean bias (P > 0.05). However, 95% LoA were smaller for the doublet (±13.9) than the twitch (±32.0). Twitch and doublet rates showed similar trends. Ratio of low (20 Hz) to high (50 Hz) frequency forces showed no mean bias (P > 0.05) and 95% LoA of (±9.2). However, there was significant mean bias (P < 0.05) and wider 95% LoA for peak force, contraction and relaxation parameters of the low and high frequency forces. In conclusion, MVC, ITT, potentiated doublet and the ratio of low to high frequency forces are recommended to most reliably examine functional muscle recovery between 2 and 72 h after damaging exercise.     

Effects of intraspecific competition on growth and photosynthesis of Atriplex prostrata

We investigated the effect of intraspecific competition on growth parameters and photosynthesis of the salt marsh species Atriplex prostrata Boucher in order to distinguish the effects of density-dependent growth inhibition from salt stress. High plant density caused a reduction of 30% in height, 82% in stem dry mass, 80% in leaf dry mass, and 95% in root dry mass. High density also induced a pronounced 72% reduction in leaf area, 29% decrease in length of mature internodes and 50% decline in net photosynthetic rate. The alteration of net photosynthesis paralleled growth inhibition, decreasing from 7.6 ± 0.9 μmol CO₂ m⁻² s⁻¹ at low density to 3.5 ± 0.4 μmol CO₂ m⁻² s⁻¹ at high density, indicating growth inhibition caused by intraspecific competition is mainly due to a decline in net photosynthesis rate. Plants grown at high density also exhibited a reduction in stomatal conductance from 0.7 ± 0.1 mol H₂O m⁻² s⁻¹ at low density to 0.3 ± 0.1 mol H₂O m⁻² s⁻¹ at high density and a reduction in transpiration rate from 6.0 ± 0.3 mmol H₂O m⁻² s⁻¹ at low density to 4.3 ± 0.3 mmol H₂O m⁻² s⁻¹ at high density. Biomass production was inhibited by an increase in plant density, which reduced the rate of photosynthesis, stomatal conductance and leaf area of plants.     

Different roles of zinc plus arachidonic acid on insulin sensitivity between high fructose- and high fat-fed rats

AimsThis study was to determine the effects of zinc plus arachidonic acid (ZA) treatment on the insulin action in the specific ZA target organs using hyperinsulinemic euglycemic clamp method.Main methods18 Sprague–Dawley rats weighing ~ 130 g were divided into 3 groups of 6 rats and treated them with 1) normal rat chow, 2) high fructose (60.0%) diet only, or 3) the same fructose diet plus drinking water containing 10 mg zinc plus 50 mg arachidonic acid (AA)/L. In a separate study, male Wistar rats weighing ~ 250 g were fed normal rat chow (n = 4) or high fat (66.5%) diet with drinking water containing zero (n = 9) or 10 mg AA plus 20 mg zinc /L (n = 9). After 4 week treatment, insulin action was assessed using the hyperinsulinemic eguglycemic clamp technique.Key findingsHigh fructose feeding impaired suppression of hepatic glucose output by insulin compared to controls during the clamp procedure (4.39 vs. 2.35 mg/kg/min; p < 0.05). However, ZA treatment in high fructose-fed rats showed a significant improvement of hepatic insulin sensitivity compared to non-treatment controls (4.39 vs. 2.18 mg/kg/min; p < 0.05). Glucose infusion rates in Wistar rats maintained on a high fat diet (HFD) were significantly lower compared to control rats (22.8 ± 1.3 vs. 31.9 ± 1.4 mg/kg/min; p < 0.05). ZA treatment significantly improved (~ 43%) peripheral tissue insulin sensitivity in HFD fed animals (26.7 ± 1.3 [n = 9] vs. 22.8 ± 1.3 mg/kg/min; p < 0.05).SignificanceThese data demonstrate that ZA treatment is effective in improving glucose utilization in hyperglycemic rats receiving either a high-fructose or a high-fat diet.     

Influence of diet complexity on productive performance and nutrient digestibility of weanling pigs

The effect of diet complexity on coefficient of total tract apparent digestibility (CTTAD) and growth performance was studied in piglets from 21 to 62 d of age. There were five experimental prestarter diets (21–41 d of age) with similar net energy and total indispensable amino acids content. The negative control diet contained 400 g raw maize, 40 g fish meal (FM) and 70 g lactose (LAC)/kg and the positive control diet contained 400 g cooked maize, 100 g FM and 140 g LAC/kg. The other three diets were similar to the positive control diet but the cooked maize was substituted by raw maize or contained 40 g FM/kg or 70 g LAC/kg, respectively. Each treatment was replicated six times (six pigs per pen). For the starter period (42–62 d of age), half of the pens of each of the prestarter treatments was sorted into two groups and fed either a standard soybean meal–raw maize–lard diet or a diet with similar nutrient profile that included 200 g cooked maize, 50 g FM, 13 g LAC, 20 g soy protein concentrate and 10 g soybean oil/kg in substitution of lower cost ingredients. Dietary treatment did not affect piglet performance at any age, but incidence of diarrhoea during the prestarter period, was higher in piglets fed the negative control diet than in piglets fed any of the other diets (P<0.05). At 30 d of age (prestarter period), the CTTAD of organic matter and gross energy were lower (P<0.001) for pigs fed the negative control diet than for pigs fed the other diets, but that of crude protein was not affected. At 50 d of age (starter period), dietary treatment did not affect the CTTAD of any dietary component. It is concluded that the use of high levels of high quality ingredients in the diet did not improve growth performance of piglets at any age. From 21 to 41 d of age, the incidence of diarrhoea was reduced and the CTTAD of dietary components was increased when the more complex diets were fed. The inclusion of high levels of high quality ingredients in the diet to maximize performance of young pigs might not be justified under all circumstances.     

Preliminary observations on the association between serum IL-6 and hydration status and cardiovascular risk in patients treated with peritoneal dialysis

IntroductionSystemic inflammation, as defined by elevated blood IL-6, is a strong independent predictor of peritoneal dialysis (PD) patient survival. The present study has aimed to determine whether there exists a particular “phenotype” associated with high systemic IL-6 that characterizes PD patients in terms of their fluid status and cardiac parameters.MethodsFifty-seven prevalent PD patients were classified according to serum concentrations of IL-6. The degree of overhydration was assessed by bioimpedance analysis (BIA). Echocardiography and serum concentrations of NT-proBNP and troponin T were used to assess cardiovascular risk.ResultsPatients with high serum IL-6 were older, more often diabetic, treated with PD for longer, and significantly more overhydrated. There was a significant correlation between serum IL-6, hydration status (r = 0.38; p = 0.002) and serum albumin (r = −0.35; p = 0.009). Multivariate regression analysis confirmed a strong association of overhydration, hypoalbuminemia, and systemic IL-6 concentration. Patients with high IL-6 had significantly increased levels of both NT-proBNP (r = 0.36; p = 0.006) and TnT (r = 0.50; p < 0.001) in the absence of abnormalities in echocardiography.ConclusionsHigh systemic IL-6 identifies PD patients with increased cardiovascular risk that is significantly related to overhydration. Thus, the measurement of serum IL-6 may contribute to the more accurate assessment of cardiovascular status in patients undergoing PD.     

Impact of nutrients and herbivory by Eccritotarsus catarinensis on the biological control of water hyacinth,Eichhornia crassipes

Many water hyacinth infestations in South Africa are the symptom of eutrophication, and as a result, biological control of this weed is variable. This study examined the effects of herbivory by the mirid, Eccritotarsus catarinensis, on water hyacinth grown at high, medium and low nitrogen (N) and phosphorus (P) nutrient concentrations. Water nutrient concentration appears to be the overriding factor affecting plant growth parameters of water hyacinth plants—at high nutrient concentrations, leaf and daughter plant production were more than double than at low nutrient concentrations, while stem length was twice as great at high nutrient concentrations compared to low concentrations. Chlorophyll content was also twice as high at high nutrient concentrations than low concentrations. Conversely, flower production at high nutrient concentrations was less than half that at low concentrations. Herbivory by E. catarinensis did not have as great an effect on water hyacinth vigour as nutrient concentration did, although it significantly reduced the production of daughter plants by 23 ± 9%, the length of the second petiole by 13 ± 5%, and chlorophyll content of water hyacinth leaves by 15 ± 6%. In terms of insect numbers, mirids performed better on plants grown under medium nutrient conditions (99 ± 28 S.E.), compared to high nutrient concentrations (52 ± 27 S.E.), and low nutrient concentrations (25 ± 30 S.E.). Thus, these results suggest that the fastest and most significant reduction in water hyacinth proliferation would be reached by lowering the water nutrient concentrations, and herbivory by E. catarinensis alone is not sufficient to reduce all aspects of water hyacinth vigour, especially at very high nutrient concentrations.     

Molecular cloning and high-level expression of a β-galactosidase gene from Paecilomyces aerugineus in Pichia pastoris

A β-galactosidase gene (designated PaGalA) was cloned for the first time from Paecilomyces aerugineus and expressed in Pichia pastoris under the control of the AOX1 promoter. The coding region of 3036 bp encoded a protein of 1011 amino acids with a deduced molecular mass of 108.7 kDa. The PaGalA without the signal peptide was cloned into a vector pPIC9K and was expressed successfully in P. pastoris as active extracellular β-galactosidase. The recombinant β-galactosidase (PaGalA) was secreted into the medium at an extremely high levels of 22 mg ml⁻¹ having an activity of 9500 U ml⁻¹ from high density fermentation culture, which is by far the highest yield obtained for a β-galactosidase. The purified enzyme with a high specific activity of 820 U mg⁻¹ had a molecular mass of 120 kDa on SDS-PAGE. PaGalA was optimally active at pH 4.5 and a temperature of 60 °C. The recombinant β-galactosidase was able to hydrolyze lactose efficiently at pH 5.0 and 50 °C. It also possessed transglycosylation activities at high concentrations of lactose. PaGalA exhibited better lactose hydrolysis efficiency in whey than two other widely used commercial lactases. The extremely high expression levels coupled with favorable biochemical properties make this enzyme highly suitable for commercial purposes in the hydrolysis of lactose in milk or whey.     

Dietary boron does not affect tooth strength,micro-hardness,and density,but affects tooth mineral composition and alveolar bone mineral density in rabbits fed a high-energy diet

The objective of this study was to determine whether dietary boron (B) affects the strength, density and mineral composition of teeth and mineral density of alveolar bone in rabbits with apparent obesity induced by a high-energy diet. Sixty female, 8-month-old, New Zealand rabbits were randomly assigned for 7 months into five groups as follows: (1) control 1, fed alfalfa hay only (5.91 MJ/kg and 57.5 mg B/kg); (2) control 2, high energy diet (11.76 MJ and 3.88 mg B/kg); (3) B10, high energy diet + 10 mg B gavage/kg body weight/96 h; (4) B30, high energy diet + 30 mg B gavage/kg body weight/96 h; (5) B50, high energy diet + 50 mg B gavage/kg body weight/96 h. Maxillary incisor teeth of the rabbits were evaluated for compression strength, mineral composition, and micro-hardness. Enamel, dentin, cementum and pulp tissue were examined histologically. Mineral densities of the incisor teeth and surrounding alveolar bone were determined by using micro-CT. When compared to controls, the different boron treatments did not significantly affect compression strength, and micro-hardness of the teeth, although the B content of teeth increased in a dose-dependent manner. Compared to control 1, B50 teeth had decreased phosphorus (P) concentrations. Histological examination revealed that teeth structure (shape and thickness of the enamel, dentin, cementum and pulp) was similar in the B-treated and control rabbits. Micro CT evaluation revealed greater alveolar bone mineral density in B10 and B30 groups than in controls. Alveolar bone density of the B50 group was not different than the controls. Although the B treatments did not affect teeth structure, strength, mineral density and micro-hardness, increasing B intake altered the mineral composition of teeth, and, in moderate amounts, had beneficial effects on surrounding alveolar bone.     

The investigation of dairy industry wastewater treatment in a biological high performance membrane system

The dairy industry is generally considered to be the largest source of food processing wastewater in many countries. The highly variable nature of dairy wastewaters in terms of volumes and flowrates and in terms of high organic materials contents such as COD 921–9004 mg L⁻¹, BOD 483–6080 mg L⁻¹, TN of 8–230 mg L⁻¹ and SS of 134–804 mg L⁻¹ makes the choice of an effective wastewater treatment regime difficult. A high performance bioreactor, an aerobic jet loop reactor, combined with a ceramic membrane filtration unit, was used to investigate its suitability for the treatment of the dairy processing wastewater. The oxygen transfer rates of the bioreactor were found to be very high (100–285 h⁻¹) on the operating conditions. A loading rate of 53 kg COD m⁻³ d⁻¹ resulted in 97–98% COD removal efficiencies under 3 h hydraulic retention time. The high MLSS concentrations could be retained in the system (up to 38,000 mg L⁻¹) with the contribution of UF (ultrafiltration) unit. During the filtration of activated sludge, the fluxes decreased with increasing MLSS. Cake formation fouling was determined as dominant fouling mechanisms. The results demonstrate that jet loop membrane bioreactor system was a suitable and effective treatment choice for treating dairy industry wastewater.     

Selection of a high-affinity and in vivo bioactive ssDNA aptamer against angiotensin II peptide

Unique features of aptamers have attracted interests for a broad range of applications. Aptamers are able to specifically bind to targets and inhibit their functions. This study, aimed to isolate the high affinity ssDNA aptamers against bio-regulator peptide angiotensin II (Ang II) and investigate their bioactivity in cellular and animal models. To isolate ssDNA aptamers, 12 rounds of affinity chromatography SELEX (Systematic Evolution of Ligands by EXponential enrichment) procedure were carried out. The SPR (surface plasmon resonance) and ELONA (enzyme linked oligonucleotide assay) analysis were used to determine the affinity and specificity of aptamers. The ability of selected aptamers to inhibit the proliferative effect of Ang II on human aortic vascular smooth muscle cells (HA-VSMCs) and their performance on Wistar rat urinary system and serum electrolyte levels were investigated. Two full-length aptamers (FLC112 and FLC125) with high affinity of respectively 7.52 ± 2.44E-10 and 5.87 ± 1.3E–9 M were isolated against Ang II. The core regions of these aptamers (CRC112 and CRC125) also showed affinity of 5.33 ± 1.15E-9 and 4.11 ± 1.09E–9 M. In vitro analysis revealed that FLC112 and FLC125 can inhibit the proliferative effect of Ang II on HA-VSMCs (P < 0.05). They also significantly reduced the serum sodium level and increased the urine volume (P < 0.05). The core regions of aptamers did not show high inhibitory potential against Ang II. It can be a spotlight that ssDNA aptamers have high potential for blocking Ang II. In conclusion, it appears that the researches focusing on high affinity and bioactive aptamers may lead to excellent results in blocking Ang II activity.