The effect of polymers on the stability of microencapsulated formulations of Bacillus thuringiensis subsp. kurstaki (Bt-KD2) after exposure to Ultra Violet Radiation

We compared the protective effect of three polymers; starch, gelatin and sodium alginate (2, 3, 5%) as coating materials, on the stability of microencapsulated formulation of Bacillus thuringiensis after exposure to Ultra Violet (UV) R. Microencapsulated Spore Crystal Aggregate (SCA) formulations were prepared by the emulsion gelling method. The protective effect of polymers was evaluated by measuring spore viability. Bioassay and release tests were done on the microencapsulated formulations. Sodium alginate (5% w/w) showed the highest viabilities of 90 and 86% after exposure to Ultra Violet in long term (UVB 385 nm) and Ultra Violet in short term (UVC 254 nm) radiation, respectively, while viabilities of non-microencapsulated spores under these conditions were 40 and 50%, respectively. The crystal activity (mortality) of irradiated and non-irradiated free spore formulations on second-instar larvae of Ephestia kuehniella were 15 and 93%. However, the mortalities caused by irradiated and non-irradiated microencapsulated formulations were 70 and 80% on the 10th day of the experiment. The size range of the microcapsules was 7–20 µm while the microcapsulation efficiency was 86%. The release behaviour of microspheres conformed best to Korsmeyer–Peppas semi-empirical model with the correlation of R² = 0.98.     

Response Surface Optimization of Lyoprotectant for Lactobacillus bulgaricus During Vacuum Freeze-Drying

The individual and interactive effects of skimmed milk powder, lactose, and sodium ascorbate on the number of viable cells and freeze-drying survival for vacuum freeze-dried powder formulation of Lactobacillus bulgaricus were studied by response surface methodology, and the optimal compound lyoprotectant formulations were gained. It is shown that skim milk powder, lactose, and sodium ascorbate had a significant impact on variables and survival of cultures after freeze-drying. Also, their protective abilities could be enhanced significantly when using them as a mixture of 28% w/v skim milk, 24% w/v lactose, and 4.8% w/v sodium ascorbate. The optimal freeze-drying survival rate and the number of viable cells of Lactobacillus bulgaricus were observed to be (64.41 ± 0.02)% and (3.22 ± 0.02) × 10¹¹ colony-forming units (CFU)/g using the optimal compound protectants, which were very close to the expected values 64.47% and 3.28 × 10¹¹ CFU/g.     

Effects of milling and surfactants on suspensibility and spore viability in Paenibacillus polymyxa powder formulations

Two milling methods and five surfactants were tested for their effects on the suspensibility of a wettable powder formulation of Paenibacillus polymyxa HY96-2 (P. polymyxa original powder or PPOP). Both hammer milling and jet milling significantly improved suspensibility of PPOP (from 8.1 to 26.4% and 58.3%, respectively), and did not decrease spore viability. Five surfactants (EFW^®, D-425^®, sodium dodecyl benzene sulfonate, tea saponin and sodium lignosulphonate) were each mixed with jet milled powder (JMP). The best results were obtained with sodium lignosulphonate and D425. These gave equal improvement in suspensibility. Sodium lignosulphonate had the best biocompatibility; spore viability values of JMP mixed with this at 1, 3, 5, or 10% (w/w) were all over 95.0%. Addition of sodium lignosulphonate to hammer milled powder (HMP) or JMP at 5% (w/w) increased suspensibility from 26.4 to 38.6% and from 58.3 to 81.1%, respectively. Our results suggest that jet milling and sodium lignosulphonate can synergistically improve the suspensibility of P. polymyxa wettable powder without adverse effects on spore viability. These findings will accelerate the commercialization of P. polymyxa wettable powder and provide a basis for the development of other biopesticides.     

Optimisation of the spray drying process of formulating the post-harvest biocontrol agent Meyerozyma caribbica

In this study, conditions of the spray-drying process of Meyerozyma caribbica were optimised using response surface methodology (RSM). The effect of three parameters (protective agent, inlet air temperature and protective agent concentration) on the cell viability of Meyerozyma caribbica was evaluated. Each parameter was evaluated at three levels. All of the evaluated factors presented an effect on the viability of the control agent. According to RSM, optimal conditions include the use of trehalose with a concentration of 7.75% (w/v) and inlet air temperature of 112.5°C. The validation of the optimal spray-drying conditions allows obtaining a formulation of M. caribbica with 95.41?±?0.93% viability, 5?±?0.37% humidity and a_w of 0.33?±?0.11. Storage for six months at 4°C presented a 5% loss in cell viability.     

Targeting the click beetle Agriotes obscurus with entomopathogens as a concept for wireworm biocontrol

Applications of Metarhizium brunneum Petch (Ascomycota: Hypocreales: Clavicipitaceae) isolate LRC112 conidia caused high mortality to Agriotes obscurus L. (Coleoptera: Elateridae) click beetles in field trials. Banded conidiated rice (4.4 × 10¹⁴ conidia ha^?1) and conidia dust (5.0 × 10¹³ conidia ha^?1) resulted in 93.3 % ± 7.3 and 91.3 % ± 3.0 mortality after 18 days, while aqueous conidia suspension spray (5.0 × 10¹³ conidia ha^?1) with and without 80 g ha^?1 spinosad resulted in 68.2 % ± 17.7 and 52.6 % ± 17.4 mortality. Differences in results between 2012 and 2013 were attributed to rainfall, with pronounced effects in 2012 (rain beginning 35 h post treatment) and minimal effects in 2013 (rain beginning at 4 h). In another field experiment, beetles dosed with 1.49 × 10⁷ ± 5.08 × 10⁶ conidia per beetle retained 4.6 % of conidia after seven days while conidia viability on beetle bodies remained unchanged. The results inferred opportunities for controlling click beetles using fungal entomopathogens, and for horizontal transmission of the inoculum.     

Reducing the microcapsule diameter by micro-emulsion to improve the insecticidal activity of Bacillus thuringiensis encapsulated formulations

The emulsion/internal gelation method is highly effective to produce microcapsules of Bacillus thuringiensis (Bt) in a short time; however, it has the limitation to produce microcapsules within a wide range of diameters (1–1000?µm). The aim of this study was to reduce the range of small microcapsule diameters by using a water/corn-oil (W/CO) micro-emulsion as the dispersing medium and the mixture Tween 80–Span 80 as the surfactant. It involved the development of the W/CO micro-emulsion and the determination of the suitable agitation time to disperse the gelling medium (sodium alginate) through the micro-emulsion. A micro-emulsion formulation that allowed reduction of the microcapsule diameter was composed of 82% corn oil, 12% alginate solution and 6% surfactant mixture Tween80–Span80 (31:69). Evaluation of four dispersing times showed that 45 min was suitable to produce 75% of microcapsules of an average diameter of 3.1?±?1?µm containing the spore–protein complex (SPC) produced by Bt. Bioassays carried out at low concentrations of microencapsulated formulations of cry proteins allowed determination of how its insecticidal effect increased if the range of microcapsule diameters was reduced in the range 1–9?µm. Furthermore, the SPC formulation in alginate microcapsules showed high resistance to extreme irradiation (2.9?±?0.5?×?10⁸ erg) of a long wavelength (365?nm), which made the microencapsulated formulation profitable and of high yield since repeated applications of the biopesticide during the same harvest period may not be necessary.     

Optimization of storage condition for maintaining long-term viability of nematophagous fungus Esteya vermicola as biocontrol agent against pinewood nematode

The fungus, Esteya vermicola has been proposed as biocontrol agent against pine wilting disease caused by Bursaphelenchus xylophilus. In this study, we reported the effects of temperature and different additives on the viability and biocontrol efficacy of E. vermicola formulated by alginate-clay. The viability of the E. vermicola formulation was determined for six consecutive months at temperature ranged from ?70 to 25 °C. The fresh conidia without any treatment were used as control. Under the optimal storage conditions with E. vermicola alginate-clay formulation, the results suggested that E. vermicola alginate-clay formulation with a long shelf life could be a non-vacuum-packed formulation that contains 2 % sodium alginate and 5 % clay at 4 °C. Three conidial formulations prepared with additives of 15 % glycerol, 0.5 % yeast extract and 0.5 % herbal extraction, respectively significantly improved the shelf life. In addition, these tested formulations retained the same biocontrol efficacy as the fresh conidial against pinewood nematode. This study provided a tractable and low-cost method to preserve the shelf life of E. vermicola.     

Liposomal amikacin dry powder inhaler: Effect of fines on in vitro performance

The aim of the present investigation was to prepare and evaluate the influence of adding fines on the in vitro performance of liposomal amikacin dry powder inhaler (AMK LDPI) formulations. Liposomes composed of hydrogenated soyaphosphatidylcholine, cholesterol and saturated soyaphosphatidylglycerol (AMK 1), or stearylamine (AMK 2) were prepared by a reverse phase evaporation technique, extruded to reduce size and separated from unentrapped drug. Purified liposomal dispersion was subjected to lyophilization using optimized cryoprotectant to achieve maximum percentage drug retentio (PDR). Lactose carrier in varying mass ratios with or without addition of fines in different mixing sequences was used to formulate AMK LDPI formulations. AMK LDPI formulations were characterized for angle of repose, compressibility index, dispersibility index, scanning electron microscopy, and fine perticle fraction (FPF). PDR was found to be 97.6%±2.2% for AMK1 and 98.5%±1.9% for AMK2 using sucrose as optimized cryoprotectant in lipid:sucrose ratio of 1∶4. Lactose carrier containing 10% fines (wt/wt) was found to be the optimum blend at 1∶5 mass ratio of liposome:lactose. The addition of fines and the order of mixing of fines were found to influence the FPF with significantly different device fractions. FPF of AMK LDPI formulations using Rotahaler as the delivery device at 30, 60, and 90 L/min were found to be 21.85%±2.2% and 24.6%±2.4%, 25.9% ±1.8% and 29.2%±2.1%, and 29.5%±2.6% and 34.2%±2.0% for AMK1 and AMK2, respectively. From the studies performed in this investigation, it was observed that liposomal charge, addition of fines and order of mixing fines, has a significant effect (P<.05) on in vitro deposition of drug from LDPI formulation.     

Effects of Metarhizium anisopliae on Meccus pallidipennis (Hemiptera: Reduviidae) over different types of wall surfaces

The entomopathogenic fungus Metarhizium anisopliae is virulent for the insect triatomine Meccus pallidipennis. To evaluate the functionality of a fungal formulation (vegetable oil and emulsifiers) of this fungus, virulence was assayed by insect mortality on the pronotum of third instar nymphs (N3) M. pallidipennis in laboratory conditions and ST₅₀ was calculated. Mortality was evaluated directly: 100%, 97.33% and 98.66% mortalities were caused by formulation, emulsified formulation and fungal conidia, respectively, at day 8 of insect infection. Another bioassay was carried out in simulated external conditions (peridomicility) using red and gray brick walls, a stone fence and mountain soil (experimental units). These simulated conditions were infected with 10?ml of a 1?×?10⁹?conidia/ml emulsified formulation by means of a manual sprinkler prior to the placement of the nymphs. Ten N3 M. pallidipennis were deposited in each experimental unit and insect mortality was monitored every 12?h for 22 days. Each treatment was replicated four times. With the red brick wall, a mortality of 90% at day 22 and a ST₅₀ of 15 days were obtained on N3 M. pallidipennis; with the gray brick wall, 100% mortality and a ST₅₀ of 13 days; and with the stone fence, 88.33% mortality and a ST₅₀ of 21 days. The results obtained in this research work indicate that the formulation with conidia of the M. anisopliae strain EH-473/4 may be auxiliary in the development of strategies for the control of Chagas disease insect transmitters such as M. pallidipennis.     

Staining for viability testing,germination and maturation of Sambucus nigra L. pollen in vitro

Freshly released pollen of black elderberry (Sambucus nigra L.) was incubated under various culture conditions until germination was achieved. Optimal conditions for germination were determined and used for maturation of unicellular microspores in vitro. Staining with 5-diphenyltetrazolium bromide, propidium iodide and iodine potassium iodide was used to assess pollen viability, nuclear phase and maturation, respectively. The germination rate was highest when fresh pollen was agitated at 40 rpm in Petri dishes containing a liquid medium consisting of Brewbaker and Kwack salts, 15% (w/v) sucrose, 500 mg/l MES sodium salt, at pH 5.0; germination reached nearly 70% after only 1 h in culture. Under these conditions, and with addition of 200 mg/l glutamine, 260 mg/l cytidine and 500 mg/l uridine, uninucleate microspores developed into mature pollen at a 12% germination rate. Our report is the first demonstration of maturation of S. nigra microspores in vitro.     

Development and fecundity of Spodoptera exigua (Hübner) (Lepidoptera: Noctuidae) on different soybean cultivars

The life history of Spodoptera exigua was studied under laboratory conditions at temperature of 25°C ± 1°C, relative humidity (RH) of 60% ± 5% and a photoperiod of 16:8 h (Light:Dark) on nine soybean (Glycine max L.) cultivars: 032, 033, Hill, M4, M7, M9, M11, TMS and Zan. Larval and pupal development times were longest on M7 cultivar (14.09 ± 0.078 and 7.78 ± 0.090 days) and shortest on 033 cultivar (11.98 ± 0.054 and 7.033 ± 0.090 days). The longest and shortest female and male longevities were observed on Hill and Zan cultivars, respectively. The results showed that the whole life span varied from 32.80 ± 0.89 to 37 ± 0.98 days for females and from 33.3 ± 0.66 to 38.74 ± 0.95 days for males. The highest daily fecundity was obtained on M9 (102. 38 eggs), and the lowest on Hill (75.87 eggs). Mean total fecundity per female during adult life was highest on 033 (1156.20 eggs) and lowest on M7 cultivars (841.68 eggs). Clustering analysis of the biological parameters of S. exigua confirmed that 033 and Hill cultivars were the susceptible and the resistant cultivars, respectively.     

Effect of temperature,relative humidity and aphid developmental stage on the efficacy of the mycoinsecticide Mycotal® against Myzus persicae

The green peach aphid, Myzus persicae, is a major pest worldwide. An examination of the impact of temperature, relative humidity (RH) and developmental stages of M. persicae on the efficacy of the whitefly mycoinsecticide Mycotal®, based on Lecanicillium muscarium and the effects of infection on aphid fecundity was evaluated under controlled conditions. Although this fungus can be grown at a broad range of temperatures (15–30°C), the optimum temperature for control of M. persicae ranged between 20 and 30°C. L. muscarium had high efficacy as a microbial control agent against M. persicae between 55% and 90% RH. Total mortality of aphids treated with different spore dosages of L. muscarium varied according to the developmental stage: adults, fourth and third instar nymphs proved more susceptible than first instar nymphs. Although the fungus did not affect the rate of nymph production, the reproductive period of aphids significantly decreased with increasing the spore dosage. Thus, total fecundity of treated aphids was 22.6 ± 1.1 and 31.6 ± 2.4 offspring per adult at the medium (644 ± viable spore/mm²) and low (330 ± 40 viable spore/mm²) dosages, compared with 45.7 ± 4.3 offspring per untreated aphid. The results suggest that L. muscarium has the potential as a biological control agent of M. persicae.     

Development of a spore-based formulation of microbial pesticides for control of rice sheath blight

An effective formulated biopesticide for controlling sheath blight in rice was developed using three microbial antagonists (Bacillus megaterium, Bacillus subtilis and Aspergillus niger) isolated from the rice sheath. The efficiency of spore-based formulations of the above microbial antagonists was investigated and their effectiveness in controlling sheath blight was demonstrated. Application of talc-based formulations of individual antagonists and mixtures of the three antagonists as spray treatments or soil applications were effective in reducing the incidence by up to 45% at 27 days after inoculation of the pathogen of sheath blight and increased rice yield. The use of spores of a fungal antagonist (A. niger), in comparison to commonly used bacterial antagonists, is a novel feature of the present study. Optimum sporulation conditions of the antagonists for preparation of spore-based formulations and their commercially desirable features such as the ability to maintain spore viability in storage were also determined. Culturing in the synthetic replacement sporulation medium (SRSM-2) for 72 hours was the most effective for sporulation of the two bacterial antagonists while culturing in potato dextrose broth (PDB) for 7 days was the most effective for sporulation of the fungal antagonist. It was demonstrated that talc-based formulations of all antagonists, either in refrigerated storage (4°C) or at room temperature (28±2°C), were able to maintain greater spore viability over a longer period (>6 months) than spore suspensions. In view of the relatively shorter life spans of formulations based on vegetative cells, spore-based formulations have a distinct advantage in achieving longer-lasting control, especially under harsh field conditions.     

Small microcapsules of crystal proteins and spores of Bacillus thuringiensis by an emulsification/internal gelation method

This paper describes a microencapsulation process of a spore crystal aggregate produced by Bacillus thuringiensis var. kurstaki HD-1. The methodology is based on the emulsification/internal gelation method, and was implemented to produce microcapsules of small diameter (<10 μm) with the capacity to protect the spore crystal aggregate from extreme ultraviolet radiation. The diameter of microcapsules was in the range of 3.1 ± 0.2–6.8 ± 0.4 μm, which is considered adequate for biological control purposes. The protective effect of the alginate coat was verified by the remaining 60 ± 2% and 40 ± 1% of spore viability and protein activity, respectively, after UV-B radiation of 236 J, and with bioassays with Spodoptera frugiperda. It is expected that the protective effect of the alginate coat will improve the effectiveness of the Bt-HD1 formulated as small diameter microcapsules, and their yield, once they are released into the environment, will also be improved.     

In vitro lung delivery of bacteriophages KS4‐M and ΦKZ using dry powder inhalers for treatment of Burkholderia cepacia complex and Pseudomonas aeruginosa infections in cystic fibrosis

Aims: To determine the feasibility of formulating and aerosolizing powders containing bacteriophages KS4‐M and ΦKZ for lung delivery and treatment of pulmonary Burkholderia cepacia complex and Pseudomonas aeruginosa infections. Methods and Results: Endotoxin‐removed bacteriophages KS4‐M and ΦKZ were lyophilized in lactose/lactoferrin 60 : 40 w/w matrix and deagglomerated in a mixer mill (without beads) to formulate respirable powders. The powders were then aerosolized using an Aerolizer^® capsule inhaler. Mass median aerodynamic diameter (MMAD) of this inhalable aerosol was determined using Andersen cascade impactor at 60 l min^?1. Measured MMAD for both types of powders was 3·4 μm, and geometric standard deviation was 1·9–2·0. Viability of bacteriophages delivered distal to an idealized mouth‐throat replica was determined from bioassays of samples collected on filters placed after the idealized replica. As a percentage of inhaler load, amount of powder delivered distal to the mouth‐throat replica, which is a measure of lung delivery, was 33·7 ± 0·3% for KS4‐M and 32·7 ± 0·9% for ΦKZ. Titres collected downstream of the mouth throat were (3·4 ± 2·5) × 10⁶ PFU for KS4‐M with an Aerolizer capsule load of (9·8 ± 4·8) × 10⁶ and (1·9 ± 0·6) × 10⁷ for ΦKZ with an Aerolizer capsule load of (6·5 ± 1·9) × 10⁷. Conclusions: Bacteriophages KS4‐M and ΦKZ can be lyophilized without significant loss of viability in a lactose/lactoferrin 60 : 40 w/w matrix. The resulting powders can be aerosolized to deliver viable bacteriophages to the lungs. Significance and Impact of the Study: Development of lactoferrin‐based bacteriophage aerosol powders solidifies the ground for future research on developing novel formulations as an alternative to inhaled antibiotic therapy in patients with cystic fibrosis.     

Bacillus cihuensis sp. nov., isolated from rhizosphere soil of a plant in the Cihu area of Taiwan

A Gram-positive, moderately halotolerant, rod-shaped, spore forming bacterium, designated strain FJAT-14515^T was isolated from a soil sample in Cihu area, Taoyuan County, Taiwan. The strain grew at 10–35 °C (optimum at 30 °C), pH 5.7–9.0 (optimum at pH 7.0) and at salinities of 0–5 % (w/v) NaCl (optimum at 1 % w/v). The diagnostic diamino acid of the peptidoglycan of the isolated strain was meso-diaminopimelic acid and major respiratory isoprenoid quinone was MK-7. Major cellular fatty acids were anteiso-C_15:0 (40.6 %), iso-C_15:0 (20.7 %) and the DNA G+C content of strain FJAT-14515^T was 37.1 mol %. A phylogenetic analysis based on 16S rRNA gene sequences indicated that strain FJAT-14515^T belongs to the genus Bacillus, and was most closely related to the reference strains of Bacillus muralis DSM 16288^T (97.6 %) and Bacillus simplex DSM 1321^T (97.5 %). Levels of DNA–DNA relatedness between strain FJAT-14515^T and the reference strains of B. muralis DSM 16288^T and B. simplex DSM 1321^T were 27.9 % ± 3.32 and 44.1 % ± 0.57, respectively. Therefore, on the basis of phenotypic, chemotaxonomic and genotypic properties, strain FJAT-14515^T represents a novel species of the genus Bacillus, for which the name Bacillus cihuensis sp. nov. is proposed. The type strain is FJAT-14515^T (=DSM 25969^T = CGMCC 1.12697^T).     

Preparation of salidroside with n-butyl β-D-glucoside as the glycone donor via a two-step enzymatic synthesis catalyzed by immobilized β-glucosidase from bitter almonds

β-Glucosidase from bitter almonds was immobilized on epoxy group-functionalized beads for catalyzing salidroside synthesis in a two-step process with n-butyl-β-D-glucoside (BG) as the glucosyl donor. The formation of salidroside ((0.59?±?0.02) M) at a yield of 39.04%±1.25% was accomplished in 8?h by the transglucosylation of immobilized β-glucosidase at pH 8.0 and 50?°C when the ratio of BG to tyrosol was 1:2 (mol/mol). A study on the influence of different glycosyl acceptors demonstrated that the yield of the glucosylation reaction of phenylmethanol and cyclohexanol was higher than that of either phenol or cyclohexanol. This may account for the selectivity of the immobilized enzyme towards the alcoholic hydroxyl group of tyrosol in the salidroside synthesis reaction. A study on the synthesis of BG via the reverse hydrolysis of immobilized β-glucosidase showed that a yield of 78.04%±2.2% BG can be obtained with a product concentration of (0.23?±?0.015) M.     

The effects of 12-week progressive strength training on strength,functional capacity,metabolic biomarkers,and serum hormone concentrations in healthy older women: morning versus evening training

ABSTRACT

Previous findings suggest that performing strength training (ST) in the evening may provide greater benefit for young individuals. However, this may not be optimal for the older population. The purpose of this study was to compare the effects of a 12-week ST program performed in the morning vs. evening on strength, functional capacity, metabolic biomarker and basal hormone concentrations in older women. Thirty-one healthy older women (66 ± 4 years, 162 ± 4 cm, 75 ± 13 kg) completed the study. Participants trained in the morning (M) (07:30, n = 10), in the evening (E) (18:00, n = 10), or acted as a non-training control group (C) (n = 11). Both intervention groups performed whole-body strength training with 3 sets of 10–12 repetitions with 2–3 minutes rest between sets. All groups were measured before and after the 12-week period with; dynamic leg press and seated-row 6-repetition maximum (6-RM) and functional capacity tests (30-second chair stands and arm curl test, Timed Up and Go), as well as whole-body skeletal muscle mass (SMM) (kg) and fat mass (FM-kg, FM%) assessed by bioelectrical impedance (BIA). Basal blood samples (in the intervention groups only) taken before and after the intervention assessed low-density lipoprotein (LDL-C), high-density lipoprotein (HDL-C), blood glucose (GLU), triglycerides (TG), high-sensitive C-reactive protein (hsCRP) concentrations and total antioxidant status (TAS) after a 12 h fast. Hormone analysis included prolactin (PRL), progesterone (P) estradiol (ESTR), testosterone (T), follicle stimulating hormone (FSH), and luteinizing hormone (LH). While C showed no changes in any variable, both M and E significantly improved leg press (+ 46 ± 22% and + 21 ± 12%, respectively; p < 0.001) and seated-row (+ 48 ± 21% and + 42 ± 18%, respectively; p < 0.001) 6-RM, as well as all functional capacity outcomes (p < 0.01) due to training. M were the only group to increase muscle mass (+ 3 ± 2%, p < 0.01). Both M and E group significantly (p < 0.05) decreased GLU (–4 ± 6% and –8 ± 10%, respectively), whereas significantly greater decrease was observed in the E compared to the M group (p < 0.05). Only E group significantly decreased TG (–17 ± 25%, p < 0.01), whereas M group increased (+ 15%, p < 0.01). The difference in TG between the groups favored E compared to M group (p < 0.01). These results suggest that short-term “hypertrophic” ST alone mainly improves strength and functional capacity performance, but it influences metabolic and hormonal profile of healthy older women to a lesser extent. In this group of previously untrained older women, time-of-day did not have a major effect on outcome variables, but some evidence suggests that training in the morning may be more beneficial for muscle hypertrophy (i.e. only M significantly increased muscle mass and had larger effect size (M: g = 2 vs. E: g = 0.5).     

Expression of tumor necrosis factor-alpha and vascular endothelial growth factor in different zones of fetal membranes: a possible relation to onset of labor

This study aimed to explore whether the altered expression of tumor necrosis factor-alpha (TNF-α), vascular endothelial growth factor (VEGF) and apoptotic changes in mid zone (MZ) and rupture zone (RZ) of fetal membranes (FM) are regulatory mechanisms associated with labor at term. Fifteen FM specimens were collected after vaginal deliveries and 13 specimens after elective caesarian section. Histological and immunohistochemical analysis were employed. Area percent of TNF-α and VEGF immunostaining and apoptotic index (AI) were evaluated using image analysis. The statistical data revealed significantly higher area % for TNF-α, VEGF immunoexpression and AI in labor compared to non-labor specimens (p < 0.0001). There was a significantly higher percentage of TNF-α immunoexpressed area in MZ compared with RZ in both groups (p < 0.0001). VEGF expression in RZ of both groups proved nearly double or triple the area % of expression relative to MZ with highly significant difference (p < 0.0001). quantitative analysis revealed near two fold increase in the AI in RZ (13.42 % ± 1.2 in labor; 11.20 % ± 0.96 in non-labor groups) when compared to MZ (7.20 % ± 0.6 in labor; 5.08 % ± 0.76 in non-labor groups) with highly significant zonal difference (p < 0.0001). Correlation analysis revealed significant correlation between apoptotic indices and area % of TNF-α (r = 0.575, p = 0.002 in non-labor; r = 0.652, p < 0.0001 in labor) and VEGF (r = 0.795, p < 0.0001 in non-labor; r = 0.668, p < 0.0001 in labor). In conclusion, Apoptosis may be regulated by TNF-α and VEGF expression in FM at labor. MZ is a step back from RZ and could participate actively in rupture of the FM during labor. TNF-α and VEGF increase with onset of labor and differentially expressed in the RZ and the MZ. These findings call for further study with tissue cultures or animal models.     

Antifungal Activity of Nanocapsule Suspensions Containing Tea Tree Oil on the Growth of Trichophyton rubrum

The aim of this study was to evaluate, for the first time, the antifungal efficacy of nanocapsules and nanoemulsions containing Melaleuca alternifolia essential oil (tea tree oil) in an onychomycosis model. The antifungal activity of nanostructured formulations was evaluated against Trichophyton rubrum in two different in vitro models of dermatophyte nail infection. First, nail powder was infected with T. rubrum in a 96-well plate and then treated with the formulations. After 7 and 14 days, cell viability was verified. The plate counts for the samples were 2.37, 1.45 and 1.0 log CFU mL^?1 (emulsion, nanoemulsion containing tea tree oil and nanocapsules containing tea tree oil, respectively). A second model employed nails fragments which were infected with the microorganism and treated with the formulations. The diameter of fungal colony was measured. The areas obtained were 2.88 ± 2.08 mm², 14.59 ± 2.01 mm², 40.98 ± 2.76 mm² and 38.72 ± 1.22 mm² for the nanocapsules containing tea tree oil, nanoemulsion containing tea tree oil, emulsion and untreated nail, respectively. Nail infection models demonstrated the ability of the formulations to reduce T. rubrum growth, with the inclusion of oil in nanocapsules being most efficient.