Enzymatic grafting of peptides from casein hydrolysate to chitosan. Potential for value-added byproducts from food-processing wastes

Tyrosinase was used to initiate the grafting of peptides onto the amine-containing polysaccharide chitosan. Chemical evidence for covalent grafting was obtained from electrospray mass spectrometry for products formed from reactions with glucosamine (the monomeric unit of chitosan) and the model dipeptide Tyr-Ala. When this model dipeptide was incubated with tyrosinase and chitosan, there was a marked increase in the viscosity of the solution. This viscosity increase provides physical evidence that tyrosinase can initiate peptide grafting onto the chitosan backbone. A peptide-modified chitosan derivative was generated by reacting chitosan (0.32 w/v%) with acid-hydrolyzed casein (0.5 w/v %) using tyrosinase. After reaction, the peptide-modified chitosan was partially purified and dissolved in an aqueous acetic acid solution. Low concentrations of this peptide-modified chitosan were observed to confer viscoelastic properties to the solutions. Specifically they conferred high viscosities and shear thinning properties to the solutions, and solutions containing only 1 w/w % of the peptide-modified chitosan behaved as weak gels. Thus, tyrosinase provides a simple and safe way to convert food-processing byproducts into environmentally friendly products that offer useful functional properties. The selectivity of tyrosinase and the relatively high reactivity of chitosan's amines allow grafting to be performed with uncharacterized peptide mixtures present in crude hydrolysates.     

Survival of the western pond turtle (Emys marmorata) in an urban California environment

The western pond turtle Emys (formerly Clemmys) marmorata is declining throughout its range, primarily due to loss of habitat via urbanization and conversion to agriculture. Urban waterways present several important challenges to freshwater turtle populations, but they also present an opportunity to maintain declining species in a ubiquitous habitat that has high public visibility. The arboretum waterway on the University of California, Davis campus is an example of an extensively altered urban habitat that supports a relatively large E. marmorata population. Over the last 6 years, we monitored the turtle population inhabiting the arboretum waterway to determine the demographic health of the population, and the challenges and opportunities that urban environments pose for pond turtles. Since 1993, the naturally existing arboretum pond turtle population has declined by approximately 40% and has shown little natural recruitment. During this time, we also introduced 31 headstarted turtles into the arboretum. Headstarting is the process of raising juveniles in captivity until they have outgrown their period of greatest vulnerability to predators, and then releasing them into the wild. Our headstarting results demonstrate that this contentious strategy is a viable option for adding young turtles to the population, although it does not address the causes of decline. Over the course of our study, we encountered nine species of non-native turtles in the waterway, and these appear to be a serious threat to the native species. As more habitat becomes urbanized, it is increasingly important to understand how freshwater turtles, such as E. marmorata, adapt to urban waterways and the impact of non-native turtles on native turtle species. Our strong feeling is that urban waterways can provide habitat for viable populations of freshwater turtles and showcase them to the public, but both the aquatic and terrestrial habitat must be managed according to the biological requirements of individual species.     

Analysis of methoxyfenozide residues in fruits, vegetables, and mint by liquid chromatography-tandem mass spectrometry (LC-MS/MS)

Methoxyfenozide [3-methoxy-2-methylbenzoic acid 2-(3,5-dimethylbenzoyl)-2-(1,1-dimethylethyl) hydrazide; RH-2485], in the formulation of INTREPID, was applied to various crops. Analysis of methoxyfenozide was accomplished by utilizing liquid-liquid extraction and partitioning, followed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Method validations for fruits, vegetables, and mint are reported. Methoxyfenozide mean recoveries ranged from 72 to 129% over three levels of fortification. The overall average of mean recoveries is 97 +/- 10%. The limit of quantitation for fruits, artichoke, cucumber, squash, and refined sugar was 0.010 ppm, with a detection limit of 0.005 ppm. For all other crops, the limit of quantitation was 0.050 ppm, with a detection limit of 0.025 ppm. No residues were found greater than the limit of quantitation in control samples. Residues above the limit of quantitation were found in all matrices except refined sugar. Foliage (bean, beet, pea, and radish) had greater residue levels of methoxyfenozide residue than their corresponding roots or pods. Other crop matrices contained <1.0 ppm of methoxyfenozide except artichoke, which had a mean of 1.10 ppm.     

Mechanical properties of metastable glacial till before and after water‐induced compaction

The mechanical properties of metastable soils have been approached from different backgrounds by civil engineers and soil scientists, with the main focus being on water‐induced self‐compaction rates. This study was conducted to determine the rebound‐recompression properties of metastable soil before and after wetting and to study the effect of water‐induced compaction on the mechanical stability depending on the overburden pressure. Rebound and recompression properties were measured with oedometer tests using sieved sandy loam, taken from a nonweathered Saalian glacial till layer, as the metastable test soil. The mechanical stability (preconsolidation load) of the dry and wetted soil was derived mathematically from recompression data after the test soil was unloaded. Void‐ratio changes during unloading and reloading were on a low level (Δe = 0.03). Rebound and recompression rates of the dry samples increased with load, whereas the void‐ratio change was twofold higher during recompression. Rebound rates of the wetted samples tended to decrease with load, while the recompression rates remained constant. Preconsolidation load calculated for the dry samples overestimated the simulated overburden pressure, whereas the wetted samples showed a high congruence. In both cases, the point of greatest curvature estimated in the earlier step of deviation gave values that were more realistic to the loads from which the unloading started. Our results indicate that both dry compaction and water‐induced compaction induce primarily plastic deformation. Furthermore, we found that in both the dry and wetted state the mechanical stability of the soil reflecting the overburden pressure remained constant.     

Surgery and Doxorubicin in Dogs With Hemangiosarcoma

Forty-six dogs with histologically confirmed hemangiosarcoma of various locations other than skin were used in a prospective study to determine the efficacy of adjuvant doxorubicin (30 mg/m² IV q 3 weeks for 5 treatments) 10 to 14 days after the tumor was partially or completely excised. Analysis of the data included information on variables that were hypothesized to influence response to therapy, disease-free interval (DFI), or survival time (ST). Other information collected included age, gender, breed, weight, prior therapy, type of surgery, location of the primary tumor, presence of metastases, number of doses of doxorubicin, response to doxorubicin therapy (complete or partial response!, and the following histological criteria: overall differentiation, nuclear pleomorphism, percent necrosis, mitotic score, total histological score, and grade. Surgery outcome (complete versus incomplete surgical excision) markedly influenced survival times (P < .001). Twenty percent of the dogs rendered free of disease were alive at 1 year, whereas none of the dogs that had residual tumor after surgery were alive at 1 year. Most of the histological criteria (nuclear pleomorphism, mitotic score, grade, overall differentiation) had marked (P < .05), or close to marked, independent associations with ST for dogs that had complete tumor removal. Results from analysis of DFI were generally similar to those of ST in dogs with complete excision of the tumor. Twenty-seven of the 46 dogs (58.7%) had all clinical evidence of tumor successfully removed. Logistic regression analysis of surgical outcome (ability to remove all visible tumor) suggested that age of the subject was the only factor markedly influencing surgical outcome (P= .017). As age increased, the probability of success increased. Those dogs that had previous treatment for their hemangiosarcoma tended (P= .08) to have a shorter DFI and ST. Therefore, complete removal of all evidence of tumor followed by 5 doses of doxorubicin may be an effective treatment for dogs with hemangiosarcoma. Dogs that had all tumor successfully removed had a mean and median ST of 267 and 172 days, respectively. Dogs with incomplete tumor removal had a mean and median ST of 172 and 60 days, respectively. Similarly, prognostic variables such as the ability to completely excise all evidence of tumor, histological criteria, and age of the patient are potentially important prognostic variables for predicting outcome.     

Potassium budgets in grassland systems as affected by nitrogen and drainage

Abstract. The main inputs, outputs and transfers of potassium (K) in soils and swards under typical south west England conditions were determined during 1999/00 and 2000/01 to establish soil and field gate K budgets under different fertilizer nitrogen (N) (0 and 280 kg ha⁻¹ yr⁻¹) and drainage (undrained and drained) treatments. Plots receiving fertilizer N also received farmyard manure (FYM). Potassium soil budgets ranged, on average for the two years, from −5 (+N, drained) to +9 (no N and undrained) kg K ha⁻¹ yr⁻¹ and field gate budgets from +23 (+N, drained) to +89 (+N, undrained). The main inputs and outputs to the soil K budgets were fertilizer application (65%) and plant uptake (93%). Animals had a minor effect on K export but a major impact on K recycling. Nitrogen fertilizer application and drainage increased K uptake by the grass and, with it, the efficiency of K used. It also depleted easily available soil K, which could be associated with smaller K losses by leaching.     

Monoclonal Antibody C219 Immunohistochemistry Against P-Glycoprotein: Sequential Analysis and Predictive Ability in Dogs With Lymphoma

In the present study, the prevalence of positive staining for P-glycoprotein using C219 monoclonal antibody was assessed in 58 tissue samples of high-grade lymphoma from dogs before initiation of chemotherapy. Samples were also evaluated at relapse in 22 dogs, at necropsy in 34 dogs, and at all 3 times in 15 dogs. The frequency of positive staining was significantly higher than that found prior to the initiation of chemotherapy at the following times: relapse ( P = .0001), necropsy ( P < .0001), and both relapse and necropsy ( P < .001, sequential data). The frequency of positive staining prior to the initiation of chemotherapy was significantly inversely related to remission ( P < .001) and survival times ( P = .0012). Similarly, when populations below and above the median initial C219 score were compared with respect to remission and survival times, the population with scores greater than the median had significantly lower remission ( P < .001) and survival ( P = .008) times, respectively. The frequency of positive staining determined at relapse was significantly inversely related to the time from relapse to death ( P = .0102). Similarly, when populations below and above the median relapse C219 score were compared with respect to the time from relapse to death, the population with C219 scores greater than the median had a significantly lower time from relapse to death ( P = .006). It appears that this immunohistochemical methodology may be used as a predictor of remission time, survival time, and the time from relapse to death. Additional studies are required to confirm the usefulness of C219 as a true marker of P-glycoprotein and to evaluate P-glycoprotein as a useful prognostic factor in dogs with lymphoma.     

Ultrasound- guided catheter biopsy of the lower urinary tract: technique and results in 12 dogs

Ultrasound-guided catheter biopsy of lesions affecting the lower urinary tract was attempted in 12 dogs with mucosal lesions affecting the bladder [nine] or urethra (three). Histological biopsies were obtained by catheter biopsy in 10 dogs, enabling diagnosis of transitional cell carcinoma in five, papilloma in two, prostatic carcinoma in two and chronic cystitis in one. Cytological samples alone were obtained in two dogs, one of which enabled a diagnosis of transitional cell carcinoma; the other contained evidence of haemorrhage and inflammation, but squamous cell carcinoma was found in a subsequent exci-sional biopsy. Intravesicular haemorrhage after biopsy was observed ultrasonographically in two dogs. Ultrasound guidance enables accurate determination of biopsy catheter position. The size of biopsies obtained by this method may limit the accuracy of histological diagnosis.     

Interleukins: Biological Properties and Therapeutic Potential

Interleukins are biologically active glycoproteins derived primarily from activated lymphocytes and macrophages. Tremendous insight into the biochemical and biological properties of interleukins has been gained with advances in recombinant DNA technology, protein purification, and cell-culture techniques. The biological properties of interleukins include induction of T-lymphocyte activation and proliferation, augmentation of neutrophil, macrophage, and T-lymphocyte cytotoxicity, and promotion of B lymphocyte and multilineage bone marrow stem-cell precursor growth and differentiation. Interleukins may play a role in the pathogenesis of several important diseases. Interleukin therapy is likely to play an important role in the treatment of cancer, infectious diseases, and immunodeficiency syndromes.     

The effect of chronic neonatal guanethidine treatment on body composition in pigs

The effect of neonatal guanethidine administration (30 mg/kg/day) during the first 32 days of life on body composition at 20 weeks of age, was assessed in two breeds of pig which were known to differ in their sympathetic responsiveness. Guanethidine administration increased the proportion of fat and decreased the proportion and absolute weight of lean in the Pietrain breed; and it had no effect in Gloucester Old Spots. Thus it can be inferred that endogenous noradrenaline is responsible for promoting leanness in a pig breed with known sympathetic overactivity.