Heterologous expression and characterization of Choline Oxidase from the soil bacterium Arthrobacter nicotianae

In the course of a microbial screening of soil samples for new oxidases, different enrichment strategies were carried out. With choline as the only carbon source, a microorganism was isolated and identified as Arthrobacter nicotianae. From this strain, a gene coding for a choline oxidase was isolated from chromosomal DNA. This gene named codA was cloned in Escherichia coli BL21-Gold and the protein (An_CodA) heterologously overexpressed as a soluble intracellular protein of 59.1 kDa. Basic biochemical characterization of purified protein revealed a pH optimum of 7.4 and activity over a broad temperature range (15–70 °C). Specific activities were determined toward choline chloride (4.70 ± 0.12 U/mg) and the synthetic analogs bis(2-hydroxyethyl)-dimethylammonium chloride (0.05 ± 0.45 × 10^–2 U/mg) and tris-(2-hydroxyethyl)-methylammonium methylsulfate (0.01 ± 0.12 × 10^–2 U/mg). With increasing number of oxidizable groups, a significant decrease in activity was noted. Determination of kinetic parameters in atmorspheric oxygen resulted in K _M = 1.51 ± 0.09 mM and V _max = 42.73 ± 0.42 mU/min for choline chloride and K _M = 4.77 ± 0.76 mM and V _max = 48.40 ± 2.88 mU/min for the reaction intermediate betaine aldehyde respectively. Nuclear magnetic resonance spectroscopic analysis of the products formed during the enzyme reaction with choline chloride showed that in vitro the intermediate betaine aldehyde exists also free in solution.     

Development of Spray Dried Liposomal Dry Powder Inhaler of Dapsone

This investigation was undertaken to evaluate practical feasibility of site specific pulmonary delivery of liposomal encapsulated Dapsone (DS) dry powder inhaler for prolonged drug retention in lungs as an effective alternative in prevention of Pneumocystis carinii pneumonia (PCP) associated with immunocompromised patients. DS encapsulated liposomes were prepared by thin film evaporation technique and resultant liposomal dispersion was passed through high pressure homogenizer. DS nano-liposomes (NLs) were separated by ultra centrifugation and characterized. NLs were dispersed in phosphate buffer saline (PBS) pH 7.4 containing different carriers like lactose, sucrose, and hydrolyzed gelatin, and 15% l-leucine as antiadherent. The resultant dispersion was spray dried and spray dried formulation were characterized to ascertain its performance. In vitro pulmonary deposition was assessed using Andersen Cascade Impactor as per USP. NLs were found to have average size of 137 ± 15 nm, 95.17 ± 3.43% drug entrapment, and zeta potential of 0.8314 ± 0.0827 mV. Hydrolyzed gelatin based formulation was found to have low density, good flowability, particle size of 7.9 ± 1.1 μm, maximum fine particle fraction (FPF) of 75.6 ± 1.6%, mean mass aerodynamic diameter (MMAD) 2.2 ± 0.1 μm, and geometric standard deviation (GSD) 2.3 ± 0.1. Developed formulations were found to have in vitro prolonged drug release up to 16 h, and obeys Higuchi's Controlled Release model. The investigation provides a practical approach for direct delivery of DS encapsulated in NLs for site specific controlled and prolonged release behavior at the site of action and hence, may play a promising role in prevention of PCP.     

An initial comparison of tubular netting versus tie–tie methods of cultivation for Kappaphycus alvarezii (Rhodophyta, Solieriaceae) on the south coast of Rio de Janeiro State, Brazil

A new cultivation technique for Kappaphycus alvarezii was used in the Brazilian southeastern coast (23°02′25″ S and 43°53′39″ W), the tubular netting on floating rafts. The tubular net technique (TN) was compared with the current method of tie–tie (TT). After 47 days, the daily growth rate (mean ± SD of TN and TT = 3.4 ± 0.7% day⁻¹), carrageenan yield (mean ± SD of TN and TT = 36 ± 1.3%), gel strength (mean ± SD of TN and TT = 730 ± 114.5 g cm2), and viscosity (mean ± SD of TN and TT = 350 ± 60.4 cP) did not differ between the two techniques (p > 0.05). The amount of time used to plant (TT = 30.2 ± 2.3 min and TN = 13.3 ± 3.4 min) and to harvest the seedlings (TT = 36.2 ± 2.7 min and TN = 17.8 ± 3.7 min) were lower in TN (p < 0.001). It is concluded that TN was more effective than TT, the cultivation management (time rates used to plant and harvest the seedlings) was 53.6% faster, no seedlings were lost, the cost was lower, and an estimated return in 1 year of nearly 20% more than that of the TT technique.     

Growth and survival performance of the gametophyte of Gigartina skottsbergii (Rhodophyta, Gigartinales) under defined nutrient conditions in laboratory culture

Gigartina skottsbergii is a commercially important carrageenan producer that has been suffering severe extraction pressure in Chile’s Magellan Region and Cape Horn Archipelago since 1998. In order to create baseline information for its cultivation and repopulation, we studied the effects of agricultural fertilizers on growth of G. skottsbergii early developmental stages. The culture media utilized were: a) seawater + Bayfoland, b) seawater + Superphosphate, c) seawater + Urea, d) seawater + Provasoli and e) seawater as a control. The culture conditions were: a) 12L:12D photoperiod; b) temperature 8 ± 1°C and c) irradiance at 45 μmol photons m⁻² s⁻¹. After 60 days, higher relative growth rates between treatments were observed; the treatments that included Bayfoland and Provasoli showed greater growth (382 ± 55 and 378 ± 50 μm, respectively,) compared to Superphosphate (88 ± 16 μm), control (78 ± 10 μm) and Urea (70 ± 11 μm) treatments, after 81 days. The Urea treatment and the control had inhibitory effects on G. skottsbergii germlings growth and survival, as evidenced by progressive loss of pigmentation and death after 60 days. These results showed that Bayfoland was an excellent alternative to develop cultures.     

Supplementary feeding during the chick-rearing period is ineffective in increasing the breeding success in the bearded vulture (Gypaetus barbatus)

One of the most widespread tools in the conservation of scavenger species is the provision of supplementary food. However, scientific studies on its effectiveness have been rarely conducted. Here, we present the first results of an experimental specific supplementary feeding programme applied from hatching to 45–60 days, aimed at increasing the breeding success of an obligate brood reducer, the threatened bearded vulture Gypaetus barbatus. We tested its effectiveness in the Spanish Pyrenees, the locale of the most important population of this species in the European Union, in which a regressive trend in breeding parameters has been observed in recent years. We compared the breeding success in nests with supplementary food to non-supplemented control nests. Supplementary food did not significantly increase global breeding success (supplemented nests, 0.793 ± 0.193 chicks per pair with eggs hatched vs non-supplemented nests, 0.771 ± 0.185) or the individual breeding performance of the territories (supplemented period, 0.712 ± 0.307 vs non-supplemented period, 0.642 ± 0.311). The similar values obtained suggest that the specific supplementary feeding programme applied during the chick-rearing period is apparently ineffective at increasing breeding success. The results suggest that, at least in the study area, factors that provoke breeding failure after hatching continue to operate independently of the supplementary feeding programme. Although more research on this subject is required, these preliminary conclusions should be taken into account by managers in order to optimise the investment of economic resources and to better prioritise the future establishment of alternative conservation actions.     

Effects of season on sleep and skin temperature in the elderly

The effects of season on sleep and skin temperature (Tsk) in 19 healthy, elderly volunteers were investigated. Measurements were obtained in summer, winter, and fall, and activity levels were monitored using a wrist actigraph system for five consecutive days. The temperature and humidity of the bedrooms of the subjects’ homes were measured continuously for five days. During actigraphic measurement, Tsk during sleep was measured for two nights. The bedroom temperature and humidity significantly increased in summer compared to winter and fall. In summer, the total sleep time decreased (mean ± SE min; summer, 350.8 ± 15.7; winter, 426.5 ± 14.2; fall, 403.2 ± 16.4) and wakefulness increased (P < 0.003) compared to those in fall or winter. The sleep efficiency index that was derived from wrist actigraphy was significantly decreased (P < 0.001) in summer (81.4 ± 2.9%) compared with winter (91.6 ± 1.3%) or fall (90.2 ± 1.2%). The forehead Tsk significantly increased, while the chest and thigh Tsks were decreased in summer compared to those in fall or winter. These results suggest that, in the elderly, sleep is disturbed in summer more than in other seasons, and that this disturbance is related to fluctuations in Tsk.     

Some reproductive characteristics of endangered Caspian Lamprey (<Emphasis Type="Italic">Caspiomyzon wagneri</Emphasis> Kessler, 1870) in the Shirud River southern Caspian Sea,Iran

Migration and reproduction of the Caspian Lamprey, Caspiomyzon wagneri, in the Shirud River were investigated during late-March to early-May at water temperatures ranging from 11 to 21.25°C. We examined the effect of water temperature on timing of spawning migrations. There was a significant negative relationship between temperature and intensive migration of Caspian Lamprey (p < 0.05). The most intensive migration of lampreys was at night (21:00–3:00 h) and when the water temperatures averaged 16°C (34.43%). The overall sex ratio (male to female) was 1.07 to 1. The individual absolute fecundity was 31 ‘758–51’ 198 eggs (mean±SD—41,924 ± 5,382). The egg diameter was 0.780–1.151 (0.92 ± 0.081) mm. The individual relative fecundity varies from 80.3 to 148.1 (107.2 ± 15.1) eggs per 1 mm of length and from 260.8 to 677.4 (397.6 ± 93) eggs per 1 g of weight. The gonadosomatic index (GSI) of females was 5.83–31.44 (11.22 ± 4.30).     

Morphological and Genetic Characterization of <Emphasis Type="Italic">Saimiri boliviensis</Emphasis>

The taxonomy of Saimiri is controversial because morphological characteristics, traditionally used for identification, are insufficient to distinguish species and subspecies. Genetic studies of specimens become relevant for captive management, especially considering their frequently unknown geographical origin. We analyzed phenotypic and genetic parameters in Saimiri spp. in Argentinean zoological gardens and biological stations to provide a more accurate taxonomic identification. We studied 27 males and 19 females of Saimiri spp. The cytogenetic analysis in mitotic metaphases corroborated a modal number of 2N = 44, XX/XY, and FN = 75 for males and FN = 76 for females. G- and C-bands, fluorescence in situ hybridization (FISH) and the pelage coloration pattern of all the specimens corresponded to Saimiri boliviensis boliviensis. We characterized for the first time the sperm cell morphology and morphometry (mean ± SE): total length: 71.39 ± 5.40 μm; head length: 5.71 ± 0.81 μm; head width: 3.76 ± 0.70 μm; acrosome length: 3.70 ± 0.82 μm; midpiece length: 12.20 ± 2.22 μm. Researchers can use the characterization of the sperm morphology as another parameter for taxonomic identification that, together with cytogenetic and molecular ones, would allow a more precise identification of individual Saimiri boliviensis boliviensis.     

Behavior of Freezable Bound Water in the Bacterial Cellulose Produced by Acetobacter xylinum: An Approach Using Thermoporosimetry

The aim of the study is to examine thermal behavior of water within reticulated structure of bacterial cellulose (BC) films by sub-ambient differential scanning calorimetry (DSC). BC films with different carbon source, either manitol (BC (a)) or glycerol (BC (b)), were produced by Acetobacter xylinum using Hestrin and Shramm culture medium under static condition at 30 ± 0.2°C for 3 days. BC samples were characterized by electron scanning microscopy and X-ray diffraction spectroscopy. The pore analysis was done by B.H.J. nitrogen adsorption. The pre-treated with 100% relative humidity, at 30.0 ± 0.2°C for 7 days samples were subjected to a between 25 and −150°C-cooling–heating cycle of DSC at 5.00°C/min rate. The pre-treated samples were also hydrated by adding 1 μl of water and thermally run with identical conditions. It is observed that cellulose fibrils of BC (a) were thinner and reticulated to form slightly smaller porosity than those of BC (b). They exhibited slightly but non-significantly different crystalline features. The freezable bound water behaved as a water confinement within pores rather than a solvent of polymer which is possible to use thermoporosimetry based on Gibb–Thomson equation to approach pore structure of BC. In comparison with nitrogen adsorption, it was found that thermoporosimetry underestimated the BC porosity, i.e., the mean diameters of 23.0 nm vs. 27.8 nm and 27.9 nm vs. 33.9 nm for BC (a) and BC (b), respectively, by thermoporosimetry vs. B.H.J. nitrogen adsorption. It may be due to large non-freezable water fraction interacting with cellulose, and the validity of pore range based on thermodynamic assumptions of Gibb–Thomson theory.     

Retinoylation Reactions are Inversely Related to the Cardiolipin Level in Testes Mitochondria from Hypothyroid Rats

The effect of hypothyroidism, induced by 6-n-propyl-2-thiouracil (PTU) administration to rats, on the retinoylation reaction and oxidative status was investigated in rat-testes mitochondria. In hypothyroid mitochondria, when compared to euthyroid controls, we found a noticeable increase in the amount of all-trans-retinoic acid (atRA) bound to mitochondrial proteins by an acylation process (34.2 ± 1.9 pmoles atRA/mg protein/360 min and 22.2 ± 1.7 pmoles atRA/mg protein/360 min, respectively). This increase, which was time- and temperature-dependent, was accompanied by a strong reduction in the cardiolipin (CL) amount in the mitochondrial membranes of hypothyroid (2.6 ± 0.2%) as compared to euthyroid rats (4.5 ± 0.5%) Conversely, a decreased retinoylation reaction was observed when CL liposomes were added to mitochondria or mitoplasts from both euthyroid and hypothyroid rats, thus confirming a role of CL in the retinoylation process. In mitochondria from the latter animals an increase of the level of oxidized CL occurred. The ATP level, which was reduced in hypothyroid mitochondria (27.3 ± 4.1 pmoles ATP/mg protein versus 67.1 ± 8.3 pmoles ATP/mg protein of euthyroid animals), was surprisingly increased in mitochondria by the retinoylation reaction in the presence of 100 nM atRA (481.5 ± 19.3 pmoles ATP/mg protein of hypothyroid animals versus 84.7 ± 7.7 pmoles ATP/mg protein of euthyroid animals). Overall, in hypothyroid rat-testes mitochondria the increase in retinoylation activity correlates with a significant depletion of the CL level, due to a peroxidation of this lipid. In addition, an enhanced production of reactive oxygen species was observed.     

Cultivation of the green alga, Codium fragile (Suringar) Hariot, by artificial seed production in Korea

Codium fragile (Suringar) Hariot is an edible green alga farmed in Korea using seed stock produced from regeneration of isolated utricles and medullary filaments. Experiments were conducted to reveal the optimal conditions for nursery culture and out-growing of C. fragile. Sampling and measurement of underwater irradiance were carried out at farms cultivating C. fragile at Wando, on the southwestern coast of Korea, from October 2004 to August 2005. Growth of erect thalli and underwater irradiance were measured over a range of depths for three culture stages. During the nursery cultivation stage (Stage I), growth rate was greatest at 0.5 m depth (0.055 ± 0.032 mm day⁻¹), where the average midday irradiance over 60 days was 924 ± 32 μmol photons m⁻² s⁻¹. During the pre-main cultivation stage (Stage II), the greatest growth rate occurred at a depth of 2 m (0.113 ± 0.003 mm day⁻¹) with an average irradiance of 248 ± 116 μmol photons m⁻² s⁻¹. For the main cultivation stage (Stage III) of the alga, thalli achieved the greatest increase in biomass at 1 m depth (7.2 ± 1.0 kg fresh wt m⁻¹). These results suggest that optimal growth at each cultivation stages of C. fragile could be controlled by depth of cultivation rope.     

Rice germ oil as multifunctional excipient in preparation of self-microemulsifying drug delivery system (SMEDDS) of tacrolimus

Surmounting the constraints of limited solubilization efficiency and prime requisite of antioxidant for conventional lipid formulations, the research work explores an edge over formulation utilizing potential applicability of rice germ oil (RGO) as a multifunctional excipient. Self-microemulsifying drug delivery system (SMEDDS) of tacrolimus (TAC) was formulated with RGO, an indigenous source of gamma-oryzanol. Being the same biological source, RGO and rice bran oil (RBO) were compared and it was found that RGO have more solubilization potential for TAC (2.2-fold) as well as higher antioxidant activity (8.06-fold) than the RBO. TAC-SMEDDS was prepared using RGO/Capmul PG8 (2:3) as an oil phase, Cremophore EL as a surfactant, and Transcutol P as a cosurfactant. The approximate particle size of TAC-SMEDDS was found to be 38 nm by dynamic light scattering and 12 nm by small angle neutron scattering. The in vitro dissolution studies showed complete and rapid drug release in 30 min compared to a plain drug (<5%) and marketed capsule (<50%). AUC and C _max were found to be 45.05 ± 15.64 ng h/ml and 3.91 ± 1.2 ng/ml for TAC-SMEDDS, 12.59 ± 5.54 ng h/ml and 0.48 ± 0.12 ng/ml for plain TAC, and 30.23 ± 10.34 ng h/ml and 2.31 ± 0.68 ng/ml for marketed formulation, respectively. The improved pharmacokinetic profile of TAC-SMEDDS is correlating to the dissolution results. Thus, gamma-oryzanol-enriched RGO acts as a potential multifunctional excipient for lipid formulations.     

Microcapsules and Transdermal Patch: A Comparative Approach for Improved Delivery of Antidiabetic Drug

Glibenclamide (GL)-loaded microcapsules (MC) and transdermal patches (TDP) were formulated and in vitro and in vivo parameters compared to find out the best route of drug administration. The formulation TDP1 having a drug–polymer ratio 1:1 showed comparatively higher GL release and better permeation across mice skin (p < 0.05). From the comparative study, it was concluded that the transdermal system of GL produced better improvement compared to oral microcapsule administration (p < 0.05). The transdermal system exhibited comparatively slow and continuous supply of GL at a desired rate to systemic circulation avoiding metabolism, which improved day-to-day glycemic control in diabetic subjects. Transdermal system of GL exhibited better control of hyperglycemia and prolonged plasma half-life by transdermal systems (9.6 ± 1.2 h) in comparison with oral microcapsule (5.84 ± 2.1 h), indicating that the drug, when administered by transdermal systems, will remain in the body for a longer period. From the glucose tolerance test, transdermal route effectively maintained the normoglycemic levels in contrast to the oral group (MC1), which produced remarkable hypoglycemia ranging from −12.6 ± 2.1% to −18 ± 2.3%. The significantly high (p < 0.05) area under the curve values observed with transdermal system (1,346.2 ± 92.3 ng ml⁻¹ h⁻¹) also indicate increased bioavailability of the drug from these systems compared to the oral route (829.8 ± 76.4 ng ml⁻¹ h⁻¹).     

Recruitment of the amphidromous goby <Emphasis Type="Italic">Sicyopterus japonicus</Emphasis> to the estuary of the Ota River,Wakayama, Japan

Ecological aspects of recruitment in the amphidromous goby, Sicyopterus japonicus, were studied from larval collections made with a set net in the estuary of the Ota River, Wakayama, Japan. The abundance patterns of the 12,766 larvae collected from 18 April to 26 August 2006 showed several peaks during the recruitment season. Their body sizes at recruitment ranged from 23.5 to 30.0 mm standard length (mean ± SD, 26.3 ± 1.1 mm), 0.11 to 0.49 g body weight (0.22 ± 0.05 g), and 8 to 20 condition factor (11 ± 2). The standard length of the goby larvae tended to decrease with the season, while their body weight slightly increased and resulted in an increase in condition factor. The recruitment of larvae occurred mainly during the daytime. Otolith growth increment analysis of 30 larvae collected by a square lift net on 30 April 2005 revealed that the oceanic larval duration after downstream migration ranged from 173 to 253 days (208 ± 22) after hatching. A limited time of recruitment in early summer and a considerably long duration of oceanic life (about a half year) appeared to be unique characteristics of this Sicyopterus species that lives in a temperate region in comparison to other tropical species of the genus Sicyopterus that all have year-round recruitment.     

Characteristics of Se-Enriched Mycelia by Stropharia rugoso-annulata and its Antioxidant Activities in vivo

Selenium (Se) is an essential trace element for humans and animals. Stropharia rugoso-annulata is a nutritional and functional mushroom containing many kinds of bioactive ingredients. The aims of this study were to investigate the Se-enrichment characteristics of S. rugoso-annulata in submerged culture and evaluate the antioxidant activities of Se-enriched mycelia in vivo in terms of the values of glutathione peroxidase (GSH-Px), superoxide dismutase (SOD), and malondialdehyde (MDA). The optimum parameters of Se-enrichment under the optimal Se concentration (150 μg/mL) in media were as follows: biomass 8.11 ± 0.25 g/L, Se content in mycelia 4,727.68 ± 13 μg/g, Se-accumulated rate 24.68 ± 1.67%, and percentage of organic Se 96.27 ± 3.26%. The mainly subsistent forms of selenium in Se-enriched mycelia were selenoprotein and selenium-polysaccharide. The contents of total amino acids (TAA) and essential amino acids (EAA) in Se-enriched mycelia were increased by 13.5 ± 1.09% and 12.8 ± 0.89%, respectively. It was efficient for Se-enriched mycelia to elevate GSH-Px and SOD activities and decrease MDA content. These results indicated that Se-enriched mycelia of S. rugoso-annulata represent a novel dietary source of bioavailable supplemental selenium.     

Effect of seawater temperature on the productivity of <Emphasis Type="Italic">Laminaria japonica</Emphasis> in the Uwa Sea,southern Japan

Recent studies on global climate change report that increase in seawater temperature leads to coastal ecosystem change, including coral bleaching in the tropic. In order to assess the effect of increased seawater temperature on a temperate coastal ecosystem, we studied the inter-annual variation in productivity of Laminaria japonica using long-term oceanographic observations for the Uwa Sea, southern Japan. The annual productivity estimates for L. japonica were 2.7 ± 2.5 (mean ± SD) kg wet wt. m⁻¹ (length of rope) (2003/2004), 1.0 ± 0.6 kg wet wt. m⁻¹ (2004/2005) and 12.1 ± 12.5 kg wet wt. m⁻¹ (2005/2006). Our previous study using the same methodology at the same locality reported that the productivity was estimated for the 2001/2002 (33.3 ± 15.2 kg wet wt. m⁻¹) and 2002/2003 (34.0 ± 8.7 kg wet wt. m⁻¹) seasons. Productivity in 2003/2004 and 2004/2005 was significantly lower than in years 2001/2002, 2002/2003 and 2005/2006. A comparison of oceanographic conditions among the 5 years revealed the presence of threshold seawater temperature effects. When the average seawater temperature during the first 45 days of each experiment exceeded 15.5°C, productivity was reduced to about 10 % of that in cooler years. Moreover the analysis of growth and erosion rates indicates that when the seawater temperature was over 17.5°C, erosion rate exceeded growth rate. Thus, an increase of seawater temperature of just 1°C during winter drastically reduces the productivity of L. japonica in the Uwa Sea.     

Anaesthesia of free-ranging Northern chamois (<Emphasis Type="Italic">Rupicapra rupicapra</Emphasis>) with xylazine/ketamine and reversal with atipamezole

One-hundred and fifty-five free-ranging Northern chamois (Rupicapra rupicapra) were anaesthetised in the course of a restocking programme using xylazine plus ketamine. Mean ± SD dosages for xylazine and ketamine were 1.9 ± 0.5 and 2.2 ± 0.7 mg/kg, respectively. In 57 chamois, sedation was reversed using 0.3 ± 0.1 mg/kg atipamezole. Although all the anaesthetic dosages tested immobilised free-ranging Northern chamois, shorter induction times (4.8 ± 2.6 min), deeper sedation with no reaction to handling in >90% of the animals and quick reversal (4.0 ± 2.7 min) were obtained using 2.5 mg/kg xylazine plus 3.0 mg/kg ketamine reversed with 0.25 mg/kg atipamezole. Under the conditions of this study, suggested standard doses are 63 mg/animal xylazine plus 76 mg/animal ketamine reversed by 6.3 mg/animal atipamezole. This anaesthetic protocol improves the results from the previous study of Dematteis et al. (Vet Rec 163:184–189, 2008) using xylazine alone.     

Brain acetylcholinesterase activity of the American pronghorn antelope <Emphasis Type="Italic">(Antilocapra americana)</Emphasis> collected from the US Army Dugway Proving Ground,Utah

Brain tissue was analyzed for acetylcholinesterase (AChE) activity from 24 American pronghorn antelope (Antilocapra americana) collected on the US Army Dugway Proving Ground (DPG) (latitude 40°13' 52" N, longitude 112°45' 01" W), Tooele County, Utah. Pronghorn antelope from DPG were evaluated against 26 pronghorn antelope collected in Wyoming. The mean AChE activity was significantly greater (P < 0.001) in the Wyoming control group (4.612 ± 0.193 μM/gm brain tissue/min) relative to DPG (4.032 ± 0.621 μM/gm brain tissue/min). The DPG database exhibited a fourfold greater coefficient of variation, a tenfold greater variance, and a threefold increase in the standard deviation when compared to control AChE activity. Furthermore, the 95% confidence interval for the control and for the DPG data were not overlapping; the entire control data set was greater than the mean DPG AChE activity. A post hoc sequential Bonferroni statistical procedure showed two significantly (P < 0.001) distinct subsets in the DPG data. Mean DPG Subset I AChE activity (4.528 ± 0.347 μM/gm brain tissue/min) was indistinct from the mean control AChE value (4.612 ± 0.193 μM/gm brain tissue/min). The mean DPG Subset II AChE activity (3.537 ± 0.387 μM/gm brain tissue/min) differed significantly (P < 0.001) from the mean control AChE activity. The sum of resulting α values from the multiple statistical tests did not exceed the selected α value of P < 0.05, validating the post hoc sequential Bonferroni statistical procedure. Pronghorn antelope represented by Subset II, experienced a 23.3% mean loss of AChE activity suggesting sub-lethal organophosphate (OP) exposure rather than a low level chronic environmental influence was experienced by a population subset of the DPG pronghorn antelope herd. The origin of the DPG sublethal OP exposure and its long-term effects are speculative.     

In Vitro Percutaneous Permeation and Skin Accumulation of Finasteride Using Vesicular Ethosomal Carriers

In order to develop a novel transdermal drug delivery system that facilitates the skin permeation of finasteride encapsulated in novel lipid-based vesicular carriers (ethosomes)finasteride ethosomes were constructed and the morphological characteristics were studied by transmission electron microscopy. The particle size, zeta potential and the entrapment capacity of ethosome were also determined. In contrast to liposomes ethosomes were of more condensed vesicular structure and they were found to be oppositely charged. Ethosomes were found to be more efficient delivery carriers with high encapsulation capacities. In vitro percutaneous permeation experiments demonstrated that the permeation of finasteride through human cadaver skin was significantly increased when ethosomes were used. The finasteride transdermal fluxes from ethosomes containing formulation (1.34 ± 0.11 μg/cm²/h) were 7.4, 3.2 and 2.6 times higher than that of finasteride from aqueous solution, conventional liposomes and hydroethanolic solution respectively (P < 0.01).Furthermore, ethosomes produced a significant (P < 0.01) finasteride accumulation in the skin, especially in deeper layers, for instance in dermis it reached to 18.2 ± 1.8 μg/cm². In contrast, the accumulation of finasteride in the dermis was only 2.8 ± 1.3 μg/cm² with liposome formulation. The study demonstrated that ethosomes are promising vesicular carriers for enhancing percutaneous absorption of finasteride.     

Effect of eight benthic diatoms as feed on the growth of red abalone (Haliotis rufescens) postlarvae

The growth rate of abalone post larvae of Haliotis rufescens fed ad libitum with a benthic monoalgal diatom culture maintained as monocultures on a semi-commercial scale, was evaluated and correlated with the biochemical composition of the diatoms. The cell size (7.0 × 4.0 μm to 21.0 × 7.5 μm), protein percentage (7.42% to 13.66%), and ash content (49.03% to 59.61%) were different among diatom strains; lipid percentage, nitrogen free extract, and energy content (Kcal g⁻¹) were similar among diatom strains. The values of essential and non-essential amino and fatty acids composition differed among diatom strains. Differences in the abalone shell length and orthogonal analyses revealed postlarval growth was dependent on the quality of the food source. Postlarvae abalone displaying the longest shell lengths were fed Nitzschia thermalis var. minor and Amphiprora paludosa var. hyalina (1,712.0 ± 61 μm and 1,709 ± 67 μm, respectively), followed by Navicula incerta (1,413.3 ± 43 μm). The fatty acid content of benthic diatoms and abalone growth rate were not correlated.