Development of new fungicides against Magnaporthe grisea: synthesis and biological activity of pyrazolo[3,4-d][1,3]thiazine,pyrazolo[1,5-c][1,3,5]thiadiazine,and pyrazolo[3,4-d]pyrimidine derivatives

Some pyrazolo[3,4-d]pyrimidin-4(5H)-thione, pyrazolo[3,4-d][1,3]thiazin-4-one/thione, and pyrazolo[1,5-c][1,3,5]thiadiazine-4-one/thione derivatives were synthesized and screened for antifungal activity against the causal agent of rice blast disease, Magnaporthe grisea. In all cases a remarkable inhibition of fungal growth was found in the range from 10 to 200 microg x mL(-1). Several compounds were able to control mycelium growth at a rate of 10 microg x mL(-1), a concentration at which the reference compound tricyclazole was completely ineffective. At least in the case of the most active substance, at the same dose the growth of seedlings or cultured cells of rice was substantially unaffected. Results allowed definition of structural requirements either to maintain or to enhance mycotoxic activity.     

30,000 years of hydrothermal activity at the lost city vent field

Strontium, carbon, and oxygen isotope data and radiocarbon ages document at least 30,000 years of hydrothermal activity driven by serpentinization reactions at Lost City. Serpentinization beneath this off-axis field is estimated to occur at a minimum rate of 1.2 x 10(-4) cubic kilometers per year. The access of seawater to relatively cool, fresh peridotite, coupled with faulting, volumetric expansion, and mass wasting processes, are crucial to sustain such systems. The amount of heat produced by serpentinization of peridotite massifs, typical of slow and ultraslow spreading environments, has the potential to drive Lost City-type systems for hundreds of thousands, possibly millions, of years.     

A neutralizing antibody selected from plasma cells that binds to group 1 and group 2 influenza A hemagglutinins

The isolation of broadly neutralizing antibodies against influenza A viruses has been a long-sought goal for therapeutic approaches and vaccine design. Using a single-cell culture method for screening large numbers of human plasma cells, we isolated a neutralizing monoclonal antibody that recognized the hemagglutinin (HA) glycoprotein of all 16 subtypes and neutralized both group 1 and group 2 influenza A viruses. Passive transfer of this antibody conferred protection to mice and ferrets. Complexes with HAs from the group 1 H1 and the group 2 H3 subtypes analyzed by x-ray crystallography showed that the antibody bound to a conserved epitope in the F subdomain. This antibody may be used for passive protection and to inform vaccine design because of its broad specificity and neutralization potency.     

A selective activity-dependent requirement for dynamin 1 in synaptic vesicle endocytosis

Dynamin 1 is a neuron-specific guanosine triphosphatase thought to be critically required for the fission reaction of synaptic vesicle endocytosis. Unexpectedly, mice lacking dynamin 1 were able to form functional synapses, even though their postnatal viability was limited. However, during spontaneous network activity, branched, tubular plasma membrane invaginations accumulated, capped by clathrin-coated pits, in synapses of dynamin 1-knockout mice. Synaptic vesicle endocytosis was severely impaired during strong exogenous stimulation but resumed efficiently when the stimulus was terminated. Thus, dynamin 1-independent mechanisms can support limited synaptic vesicle endocytosis, but dynamin 1 is needed during high levels of neuronal activity.     

One- and two-dimensional electrophoretic identification of IgE-binding polypeptides of Lupinus albus and other legume seeds

The prevalence of food allergies in the world population requires integrated approaches to identify new potential allergens, especially those of plant origin. The aim of this work was the allergen in vitro analysis of Lupinus albus seed proteome, a promising food protein source, and the assessment of IgE cross-reactivities with other more diffused legume species. A combination of one- and two-dimensional gel electrophoresis and immunoblotting analyses with specific IgGs for band identification and lupin-sensitized patients' circulating IgEs for allergenicity studies has been used. Two lupin proteins, namely, conglutin gamma and 11S globulin basic subunits, strongly reacted with all patients' sera. Also, cross-reactivities with the homologous polypeptides of other legume species were observed. Otherwise, no reaction at all was detected with a 2S-type lupin protein. This global electrophoretic approach has allowed the identification of a new potential lupin allergen and confirmed the cross-reactivity among the legume 11S globulin basic subunits.     

A simple and sensitive liquid chromatography-mass spectrometry confirmatory method for analyzing sulfonamide antibacterials in milk and egg

A simple and specific method able to identify and quantify traces of 14 sulfonamide antibacterials (SAs) in milk and eggs is presented. This method uses a single solid-phase extraction (SPE) cartridge for simultaneous extraction and purification of SAs in the above matrices. Milk and egg samples are passed through a Carbograph 4 sorption cartridge. After analyte desorption, an aliquot of the final extract is injected into a liquid chromatography-mass spectrometry (LC-MS) instrument equipped with an electrospray ion source (ESI) and a single quadrupole. MS data acquisition is performed in the positive-ion mode and by a time-scheduled multiple-ion selected ion monitoring program. Compared to two published methods, the present protocol extracted larger amounts of SAs from both milk and egg and decreased the analysis time by a factor of 3 with milk samples and by a factor of 2 with egg samples. Recovery of SAs in milk at the 5 ppb level ranged between 76 and 112% with relative standard deviations (RSDs) of 相似文献   

Analysis of caprine IgG1 and IgG2 subclass responses to Chlamydia psittaci infection and vaccination

Enzyme-linked immunosorbent assays (ELISAs) specific for caprine IgG(H+L), IgG1 and IgG2 were developed and evaluated for serodiagnosis of Chlamydia psittaci infections in a Tunisian goat flock with currently occurring chlamydial abortions and a clinically inapparent goat flock of an animal research facility. Additionally, ELISAs were applied to record the IgG1 and IgG2 dynamics of four goats vaccinated with inactivated Chlamydia psittaci and Coxiella burnetii. For screening purposes, the IgG(H+L) ELISA proved to be superior to the complement fixation test because it detected a larger number of chlamydial abortions and was easier to perform and to interpret. Analysis of Chlamydia psittaci-specific IgG1 and IgG2 responses to naturally occurring infections by ELISA revealed high IgG1 levels associated with IgG2 in goats with current abortions, whereas clinically inapparent, but seropositive goats were characterized by significantly lower IgG1 levels only (P less than 0.001). Similarly, the four vaccinated goats responded initially with Chlamydia psittaci-specific IgG1, whereas second and third vaccinations induced (as in goats with chlamydial abortions) predominantly IgG1, but also IgG2. The results indicated that clinically inapparent chlamydial infection may be distinguished from overt disease by analysing specific IgG1 and IgG2 responses. Applying Coxiella burnetii- specific ELISAs on field samples, IgG1 alone could be detected in eight, IgG2 alone in one and IgG1 combined with IgG2 in nine goats. The coxiella-specific antibody response of the four vaccinated goats was--in contrast to the chlamydia-specific response--characterized by IgG2 dominance.     

Isotretinoin in the treatment of multiple benign pilomatrixomas in a mixed-breed dog

An 18-month-old cross-breed dog developed multiple skin nodules, which over 8 months had increased in size and number. Biopsy samples were submitted for histopathological examination and revealed multiple benign pilomatrixomas. The dog was treated with 1 mg kg(-1) isotretinoin daily, which led to a stabilization of the size and number of the lesions. Attempts to lower the dosage to an every other day regimen led to an increase in the nodules' size and number. Monthly complete blood count and chemistry tests and Schirmer tear tests were performed to monitor any adverse effects of the retinoid treatment. No adverse effects were noted. To the authors' knowledge, this is the first report of multiple pilomatrixomas arising in a young dog and successfully controlled with oral retinoids.     

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.