Thermodynamics of adsorption of laccaic acid onto chitosan and associated dye toxicity studies

The thermodynamics of adsorption of laccaic acid (lac dye) onto chitosan were investigated under acidic condition over various concentrations (20–293 mg/l). Langmuir, Freundlich, and Temkin isotherms were used to analyze the equilibrium data at different temperatures, with the Freundlich isotherm fitting the experimental data significantly better than the other isotherms. The effect of temperature on the adsorption isotherm was studied by carrying out a series of isotherms at 10, 20, 40, and 60 °C. It was found that more dye was strongly adsorbed by chitosan when the temperature of the dye solution increased. Thermodynamic parameters such as free energy change (ΔG°), enthalpy change (ΔH°), and entropy change (ΔS°) were also evaluated. The negative value of ΔG° and positive value of ΔH° indicated that the laccaic acid adsorption process is a spontaneous and an endothermic one. Attenuated total reflectance Fourier transform infrared spectroscopy confirmed the functional groups of chitosan that affected the laccaic acid adsorption. Therefore, laccaic acid molecules could interact with the chitosan via electrostatic forces, hydrogen bonding, and ion-dipole interactions under acidic condition. From the toxicity study, the dye solution with the initial dye concentrations of 40 and 120 mg/l before dye removal showed significant mortality to earthworm Diplocardia communis (P<0.01).     

Can traditional forest management buffer forest depletion? Dynamics of Moroccan High Atlas Mountain forests using remote sensing and vegetation analysis

On the south shore of the western Mediterranean Basin, mountain forest ecosystems are degraded, mainly due to their overexploitation. Topographic, edaphic and climatic conditions create stressful growing conditions and sensitive ecosystems. Nonetheless, in these ecosystems, forests remain an important resource for the subsistence of local populations. Historically the vulnerability of this resource has prompted mankind to establish traditional control forms of forest and pastoral areas. These common resource management systems are still functioning in the Moroccan High Atlas Mountains under the name of agdal which refers to the territory, the resources and access rules laid down by the local population in order to manage the territory.     

Fecal progesterone analysis by time-resolved fluoroimmunoassay (TR-FIA) for monitoring of luteal function in the sika doe (Cervus nippon centralis)

Fecal progesterone content was measured by time-resolved fluoroimmunoassay (TR-FIA) in the sika doe (Cervus nippon). The total recovery rate of fecal progesterone by twice extraction with diethylether was about 60%. The displacement curve of TR-FIA with serial doses of fecal extract (0.156-5.0 mg feces) was closely parallel to that of the reference standard. Fecal progesterone content was correlated with that of plasma (r=0.829, n=16), but the values were 100-fold higher in feces than in plasma. Fecal progesterone content periodically changed during the breeding season suggesting the estrous cycle in the doe. The fecal progesterone content was higher between the estruses, and decreased after estrus. The time between the onset of estrous signs and the lowest fecal progesterone content was 1-2 days suggesting the time required for hepatic metabolism and intestinal passage. Fecal progesterone content was also decreased around the time of vaginal discharge. The discharge took place within a few days, suggesting a short luteal phase. Not of all decreases in fecal progesterone values were preceded by estrous behavior or vaginal discharge. Fecal progesterone content was further increased in pregnancy rather than in the preceding estrous cycle and the levels were maintained up to term. These results suggest that fecal progesterone measurement is a useful tool for non-invasive analysis of luteal function in the sika doe. The TR-FIA kit, designed for the human hospital market, was shown to be successfully utilized for fecal assay in the sika doe with minor modifications.     

Relationships between tropomyosin and myosin heavy chain isoforms in bovine skeletal muscle

The composition of tropomyosin (TPM) and myosin heavy chain (MyHC) isoforms was analyzed in 10 physiologically different bovine muscles ( masseter , diaphragm, tongue, semispinalis, pectoralis profundus , biceps femoris, psoas major , semimembranosus, longissimus thoracis and semitendinosus ) to clarify the relationships between TPM and MyHC isoforms in different muscle fiber types. The content of TPM1 and TPM3 was different in muscles according to their function in muscle contraction, although the content of TPM2 was constantly about 50% of the total TPM in all muscles. The content of TPM1 was higher in semimembranosus , longissimus thoracis and semitendinosus, while that of TPM3 was higher in masseter and diaphragm. The high positive correlation between MyHC-slow content and TPM3 content ( r  = 0.92) suggested a coexpression of TPM3 and MyHC-slow isoforms in a muscle fiber. MyHC-slow and TPM3 were expressed at the same level in masseter and diaphragm, whereas there was more TPM3 than MyHC-slow in tongue and semispinalis , so it appears that the excess TPM3 in tongue and semispinalis is expressed with other MyHC isoforms. MyHC-2a was the only fast type isoform expressed in tongue and semispinalis . Therefore, the excess TPM3 was composed of myofibrils with MyHC-2a. The results suggested that a fiber expressing MyHC-2a would be regulated delicately by changing the TPM isoform types.     

A direct time-resolved fluoroimmunoassay (TR-FIA) for measuring plasma estradiol-17beta concentrations in cattle

A direct Time-Resolved Fluoroimmunoassay (TR-FIA) system for measuring estradiol-17beta (E(2)) in bovine plasma was developed and evaluated. A 100 microl sample of bovine plasma was used for a TR-FIA without prior extraction and purification. The dose-response curves of reference standards ranged from 0.0625 to 10 pg/well. The minimum detectable concentration of this assay system was 0.625 pg/ml, and 19 pg/ml of E(2) caused a 50% reduction of maximum binding. The intra- and inter-assay coefficients of variation were 10.2 and 17.4%, respectively. The plasma E(2) concentrations measured by direct TR-FIA correlated closely with those measured after extraction (r=0.939). The results in the present study indicate that the TR-FIA reagent for E(2), designed for human research can also be utilized, with some modification, for direct assaying in bovine plasma. This assay type seems to fulfill the requirements for safety, sensitivity, specificity, reproducibility and practical convenience.     

Production of GM-CSF Mediated by Cysteine Protease of Der f in Canine Keratinocytes

House dust mite (HDM) allergens are the most common allergens for induction of IgE-mediated hypersensitivity. Recently, epicutaneous sensitization with HDM allergens has been emphasized in the development of atopic dermatitis (AD) by producing various soluble factors in keratinocytes. Among the soluble factors, GM-CSF is a key molecule that activates Langerhans cells, antigen-presenting cells in the epidermis. In the present study, we investigated the effects of Dermatophagoides farinae (Der f) on GM-CSF production in a canine keratinocyte cell line, CPEK. CPEKs were found to produce GM-CSF upon stimulation by Der f. The GM-CSF production was suppressed by addition of a cysteine protease inhibitor. The present results suggest that cysteine protease-derived Der f may be an initiator of allergic inflammation by inducing the production of GM-CSF in keratinocytes.     

Efficacy of a surgical scrub including 2% chlorhexidine acetate for canine superficial pyoderma

The clinical efficacy of a surgical scrub containing 2% chlorhexidine acetate (2CA; Nolvasan^® Surgical Scrub; Fort Dodge Animal Health, USA) was evaluated for the topical management of canine superficial pyoderma. The first study was a randomized, double‐blind, controlled trial. The control was a shampoo containing 4% chlorhexidine gluconate (4CG; Skin Clinic Shampoo; CHD MEDICS, Goyang, Korea). Ten dogs with symmetrical lesions of canine superficial pyoderma were allocated to receive either 2CA or the control shampoo applied to either side of the body twice weekly for 1 week. Both the owners and the investigators subjectively scored skin lesions including pruritus, erythema, crusted papules and scales on a scale of 0–3. The 2CA and 4CG resulted in almost the same degree of improvement of skin lesions, and there were no significant differences between the two groups. The second study was an open trial of 2CA monotherapy in eight dogs with cefalexin‐resistant Staphylococcus intermedius group‐associated superficial pyoderma. The 2CA monotherapy was applied every 2 days for 2 weeks. Five dogs improved with 2CA monotherapy, one partially improved and two did not. No adverse reactions were seen in either trial. This suggests that a 2CA surgical scrub could be a useful and safe topical adjunct therapy for dogs with superficial pyoderma involving cefalexin‐resistant Staphylococcus intermedius group.     

Analysis of fecal nutrients and particle size in captive proboscis monkeys (Nasalis larvatus) fed seasonal dietary foliage at a Japanese zoo

One male and three female captive proboscis monkeys (Nasalis larvatus) were used as test animals to compare fecal nutrients after being fed seasonal foliage. Foliage and fecal samples were collected during three seasons (spring, summer, and winter). We analyzed crude ash (CA), ether extract (EE), crude protein (CP), neutral detergent fiber (NDF), acid detergent fiber (ADF), and acid detergent lignin (ADL). In addition to the above components, for the fecal samples, we examined total fecal nitrogen (TFN), nitrogen in fecal NDF (NDF-N), metabolic fecal nitrogen (MFN), and particle size distribution (MPS). Seasonal differences in foliage components were observed, with NDF and ADF being lowest in spring (p < 0.05). Fecal NDF and ADF also tended to be lowest in spring. Further, the distribution of fecal particle size indicated that MPS tended to increase in winter. The mean fecal NDF content (47.4%) for all three seasons measured in this study was as much as 1.3 times lower than previously reported values in free-ranging specimens. These data provide useful information that can be used to find some suitable food items for the endangered highly folivorous monkeys in captivity.     

Nucleotide sequence polymorphisms of beta1‐, beta2‐, and beta3‐adrenergic receptor genes on Jinhua,Meishan, Duroc and Landrace pigs

The full amino acid coding sequences of adrenergic receptor genes beta1, beta2, and beta3 (ADRB1, ADRB2, and ADRB3)were determined for Jinhua, Meishan, Duroc and Landrace pigs. Non‐synonymous substitution of Arg458Pro was found in the porcine ADRB1 gene, resulting in a 469 amino acid sequence. Continuous substitutions of Asn29Asp and Glu30Gln were found in the porcine ADRB2 gene, resulting in a 418 amino acid sequence. Additionally, a Lys30 polymorphism of the ADRB2 gene was found in the Jinhua pigs. There were three non‐synonymous substitutions of Asn24Thr, Arg264Gln and Asn398Asp on the porcine ADRB3 gene. A thymine insertion in the ADRB3 gene, resulting in a protein with two fewer amino acids, was found in the Jinhua and Meishan pigs. To assess the effect of ADRB polymorphisms on porcine subcutaneous fat layer thickness, we calculated the genetic frequency of the variants in fatty and lean groups, each consisting of 24 pigs that were crossbreds of Duroc and Jinhua pigs. The effect of the ADRB3 gene polymorphism was not evaluated, because there was insufficient variation on the ADRB3 gene in the examined groups. Although Fisher's exact test showed no significant difference in the frequency of ADRB1 and ADBR2 variants between the two groups, the Arg458 variant of ADRB1 was higher (P = 0.11) in the lean group, and pigs in that group had a thinner fat layer than did those with the Pro458 variant. These results imply a possibility of ADRB1 polymorphism as a minor factor in porcine fat layer thickness. The Asp29 variant of ADRB2 was higher in the lean group (P = 0.11), and the Glu30 variant was higher in the fatty group (P = 0.15), but the Asp29 variant was found only in the Chinese pigs. Thus, the effect of ADRB2 polymorphisms was not clear in this study.