Conformational states of sunflower (Helianthus annuus) Helianthinin: effect of heat and pH

The structure and solubility of helianthinin, the most abundant protein of sunflower seeds, was investigated as a function of pH and temperature. Dissociation of the 11S form (hexamer) into the 7S form (trimer) gradually increased with increasing pH from 5.8 to 9.0. High ionic strength (I = 250 mM) stabilizes the 11S form at pH > 7.0. Heating and low pH resulted in dissociation into the monomeric constituents (2-3S). Next, the 7S and 11S forms of helianthinin were isolated and shown to differ in their secondary and tertiary structure, and to have denaturation temperatures (T(d)) of 65 and 90 degrees C, respectively. Furthermore, the existence of two populations of the monomeric form of helianthinin with denaturation temperatures of 65 and 90 degrees C was described. This leads to the hypothesis that helianthinin can adopt two different conformational states: one with T(d) = 65 degrees C and a second with T(d) = 90 degrees C.     

Reconstruction of the lower eyelid by third eyelid lateral advancement and local transposition cutaneous flap after "en bloc" resection of squamous cell carcinoma in 5 cats

OBJECTIVE: To describe a novel technique for blepharoplasty to cover a tissue defect involving >/=50% of the lower eyelid. STUDY DESIGN: Prospective clinical study. ANIMALS: Five cats with lower eyelid squamous cell carcinoma (SCC). METHODS: En bloc resection of SCC by removing >/=50% of the lower lid with either the medial or lateral canthus was performed without other adjunctive treatment for SCC. The lid defect was reconstructed with a transposition skin flap derived from the frontal (medial defect) or temporal (lateral defect) region. The third eyelid was advanced laterally without dissection from its insertion; its outer conjunctival layer was removed, and the skin flap was sutured with single interrupted sutures dorsally over the nictitating membrane, ventrally to the cutaneous edge of the surgical wound and medially or laterally (depending on the canthus removed) to the skin of the remaining lower lid. RESULTS: Satisfactory cosmetic and functional results were achieved and the Schirmer tear tests were normal. In 2 cats, the skin flap needed monthly hair trimming to avoid corneal lesions. CONCLUSIONS: After en bloc resection of SCC involving >/=50% of the lower eyelid, reconstruction can be achieved by relocation of the third eyelid and use of a cutaneous transposition flap sutured to the scarified external surface of the third eyelid. Eyelid apposition and lacrimal function were preserved. CLINICAL RELEVANCE: Blepharoplasty using a cutaneous transposition flap sutured to the scarified surface of a relocated third eyelid should be considered for reconstruction of lower eyelid defects with >/=50% tissue loss of the lid margin.     

Clinical and pharmacokinetic characteristics of intracavitary administration of pegylated liposomal encapsulated doxorubicin in dogs with splenic hemangiosarcoma

BACKGROUND: Canine splenic hemangiosarcoma (HSA) is a fatal malignancy, and most affected dogs die within a few months of diagnosis. Most dogs present with signs from tumor rupture, resulting in hemoabdomen and intra-abdominal dissemination. The abdomen is also the main site of disease recurrence. HYPOTHESIS: Intraperitoneal (IP) administration of doxorubicin will delay or prevent intra-abdominal tumor recurrence and prolong survival in dogs with HSA. ANIMALS: Fourteen dogs with splenic HSA. METHODS: A prospective, unmasked, uncontrolled clinical trial. After staging of disease status and splenectomy, pegylated liposomal encapsulated doxorubicin was administered intraperitoneally (1 mg/kg body weight) every 3 weeks for 4 cycles. All dogs were monitored for recurrence of HSA. Samples of plasma and abdominal fluid were collected for measurement of doxorubicin concentration and pharmacokinetic analysis. Nonlinear mixed-effect modeling was used to describe the pharmacokinetics of liposomal doxorubicin administered IP. RESULTS: All 14 dogs died, 12 because of HSA and 2 from other causes. Postmortem examination was performed on 12 dogs. All 12 dogs died because of HSA-related causes and had hepatic metastases and hemoabdomen. The IP-treated dogs had fewer serosal, mesenteric, and omental metastases than historical controls treated with systemic doxorubicin. Results of the postmortem examination and pharmacokinetic analysis confirmed that IP delivery of doxorubicin resulted in an effective drug concentration with a clearance comparable with that after i.v. delivery. CONCLUSIONS AND CLINICAL IMPORTANCE: IP pegylated liposomal encapsulated doxorubicin administration did not prevent intraabdominal recurrence of HSA in dogs.     

Associations between bacterial genotype and outcome of bovine clinical Staphylococcus aureus mastitis

Background

Staphylococcus aureus is an important cause of clinical mastitis in dairy cows worldwide. The cure rate after antimicrobial treatment of clinical S. aureus mastitis is very variable due to both cow and bacterial factors. Studies have shown that bacterial genotype might affect short-term bacteriological and clinical cure, but the long-term outcome has been less studied. The objectives of this study were to investigate associations between bacterial genotype and long-term outcome of veterinary-treated clinical mastitis (VTCM) caused by S. aureus during a follow-up period of 120 days and to study genotype variation among Swedish S. aureus isolates. S. aureus isolates from cases of VTCM were genotyped by pulsed-field gel electrophoresis. Long-term outcome measurements used were somatic cell count (SCC), additional diagnoses of VTCM, milk yield and culling. Isolates were classified into clusters (>80% similarity) and pulsotypes (100% similarity). Clusters and pulsotypes were grouped according to occurrence. Multivariable mixed-effect linear regression models including cow and bacterial factors with possible influence on SCC or milk yield were used to calculate differences in SCC or milk yield between groups. Additional outcome measures were calculated using a test of proportions.

Results

The isolates (n = 185) were divided into 18 clusters and 29 pulsotypes. Two pulsotypes were classified as common, and were found in 64% of the cases of VTCM. Remaining isolates were classified as less common or rare pulsotypes. The distribution was similar at cluster level. Outcome was calculated from follow-up data on 111 cows. Significantly lower SCC during the follow-up period was found in cows infected with common clusters compared to in cows infected with less common/rare clusters. The proportion of cows with SCC <200 000 cells/ml during the whole follow-up period was significantly higher in the group common clusters than in the group less common/rare clusters. Bacterial genotype did not influence the other outcome parameters.

Conclusions

In Sweden, two S. aureus pulsotypes, identified in about 64% of clinical S. aureus cases, were widespread. Cows infected with the common genotypes had significantly lower SCC during 120 days after treatment compared to cows infected with less common or rare genotypes.     

Prediction of field atrazine persistence in an allophanic soil with Opus2

A modified version of the model Opus was applied to measurements of soil water dynamics and atrazine (6-chloro-N2-ethyl-N4-isopropyl-1,3,5-triazine-2,4-diamine) persistence in a Bruntwood silt loam soil (Haplic Andosol, FAO system) in Hamilton, New Zealand. The modified model, Opus2, is briefly described and parameter estimation for the simulations is discussed. Soil water dynamics were more accurately described by applying measured soil hydraulic properties than by estimating them using pedotransfer functions. A parameter sensitivity analysis revealed that degradation was the most relevant process in simulating pesticide behaviour by Opus2. The Arrhenius equation incorporated in Opus2 did not correctly describe the effect of temperature on degradation rates obtained at 10, 20 and 30 degrees C. However, as the Arrhenius coefficient is a very sensitive parameter and soil temperature variation was relatively narrow in the field, the Arrhenius coefficient was approximated from the laboratory study. The simulation results obtained were superior to modelling at constant temperature. Field measured persistence of atrazine in the topsoil was underpredicted using the half-life determined in the laboratory at 10 degrees C. Modelling with a lag phase followed by accelerated degradation by use of a sigmoidal degradation equation in Opus2 significantly improved the modelling results. Nevertheless, degradation processes in the laboratory under controlled conditions did not accurately represent field dissipation, however well the laboratory degradation data could be described by simple kinetic equations. The study indicates the importance of improving field techniques for measuring degradation, and developing laboratory protocols that yield degradation data that are more representative of pesticide dynamics in field soils.     

Therapeutic effects of systemic or intramammary antimicrobial treatment of bovine subclinical mastitis during lactation

The short- and long-term treatment efficacy of administrating penicillin for bovine subclinical mastitis during lactation when using intramuscular (IM; 9.5 mg [15,000 IU]/kg bodyweight of benzyl penicillin potassium) injections twice daily for 5 days, or intramammary (IMM; 0.3g [300,000 IU] penethamate hydroiodide) administration once daily for 5 days was compared with a control group receiving no treatment. One hundred and twenty-six cows met the inclusion criteria, which were lack of clinical symptoms, no recent treatment with antimicrobials, and findings of penicillin-sensitive Staphylococcus aureus, Streptococcus dysgalactiae, or Streptococcus uberis in combination with an inflammatory reaction. At follow-up 42-58 days after treatment, the proportion of cows negative for the original infection was significantly higher in IM and IMM groups compared to controls, but the difference between antimicrobial treatment groups was not significant. The udder quarter milk somatic cell count (SCC) was significantly lower at follow-up in IM and IMM groups than in controls, but milk production did not differ between treatments. The culling rate during the 10-month period following treatment was significantly higher in the group treated with IMM penicillin than in the other two groups, but the risk of new mastitis treatments within 10 months did not differ between the three groups. The cure rate was significantly affected by lactation number (lower in older cows), breed (lower in the Swedish Holstein breed), pathogen (lower for S. aureus), and pre-treatment SCC (higher for above average SCC). In conclusion, beneficial long-term effects of antimicrobial treatment during lactation of subclinical mastitis caused by S. aureus, Str. dysgalactiae or Str. uberis were not found in the present study.