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21.
Increased calcium (Ca) in potatoes may increase the production rate by enhancing tuber quality and storability. Additionally, increased Ca levels in important agricultural crops may help ameliorate the incidence of osteoporosis. However, the capacity to alter Ca levels in potato tubers through genetic manipulations has not been previously addressed. Here we demonstrate that potato tubers expressing the Arabidopsis H+/Ca2+ transporter sCAX1 (N-terminal autoinhibitory domain truncated version of CAtion eXchanger 1) contain up to 3-fold more Ca than wild-type tubers. The increased Ca appears to be distributed throughout the tuber. The sCAX1-expressing potatoes have normally undergone the tuber/plant/tuber cycle for three generations; the trait appeared stable through successive generations. The expression of sCAX1 does not appear to alter potato growth and development. Furthermore, increased Ca levels in sCAX1-expressing tubers do not appear to alter tuber morphology or yield. Given the preponderance of potato consumption worldwide, these transgenic plants may be a means of marginally increasing Ca intake levels in the population. To our knowledge, this study represents the first attempts to use biotechnology to increase the Ca content of potatoes.  相似文献   
22.
Rain samples were collected sequentially by amount (≈2.7 mm each) from individual events at a single, relatively isolated, suburban site from August 1977 to July 1980. Rain pH's for ≤ 3 mm samples closely fit a monomodal Gaussian distribution with a median of 4.50 and a standard deviation of 0.39. The variability in pH was primarily interevent as opposed to intraevent. The 3-yr volume-weighted pH was 4.35 ± 0.02 for 3.16 m collected; annual pH's were 4.31, 4.37, and 4.38, and cumulative H+ deposition was 141 mg H+ m?2. Event-averaged rain pH and meteorological and air quality data were correlated. Low pH was associated with low rainfall volume and rate; rain after several dry days; rains with northeast surface winds; high SO2, NO2, and O3 in the ambient atmosphere; and high, strongly correlated, SO4 = and NO3 ? rainwater concentrations. The lowest 3-yr seasonal average pH (4.31) occurred during summer; values for other seasons were ≈4.37. Average intraevent H+ molarity (volume-weighted) was accurately characterized by 6.89 E?5 *(mm ram)?0.215. The relative merits of composite (e.g., whole event) and sequential sampling are examined.  相似文献   
23.
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   
24.
On the basis of previous studies on the mechanism-based inhibition, activation, and active site structure of myrosinase(s) isolated from Sinapis alba and other cruciferous seeds, crambe myrosinase shows uncommon properties and behavior. For this reason homogeneous crambe myrosinase was isolated and investigated to establish the most important physicochemical features, including kinetic properties determined with the epimers progoitrin (R) and epi-progoitrin (S) as substrates, with and without ascorbate as an activator. The results of this study demonstrate that crambe myrosinase is highly specific for epi-progoitrin due to a better stabilization of the enzyme-substrate complex. This stabilization is caused by additional hydrogen bonding that only epi-progoitrin can set up between its hydroxyl group and a suitable residue in the hydrophobic pocket where the "docking" of the glucosinolates side chain takes place.  相似文献   
25.
OBJECTIVE: To evaluate serum titers obtained by use of the microscopic agglutination test (ie, MAT titers) to Leptospira interrogans serovar pomona and autumnalis and Leptospira kirschneri serovar grippotyphosa in dogs given a commercial vaccine against serovars pomona and grippotyphosa. ANIMALS: Forty 12-week-old puppies and 20 mature Beagles. PROCEDURE: Puppies received a commercial vaccine against serovars pomona and grippotyphosa at 12 weeks of age, then received a booster vaccine and 3 weeks later; mature dogs received the vaccine once. Serum MAT titers to serovars pomona, autumnalis, and grippotyphosa were measured before vaccination and at 2, 4, 6, 10, and 16 weeks after the first or only vaccination. RESULTS: Of the 40 puppies vaccinated, 40, 0, and 40 developed MAT titers of > 100 after vaccination to serovars pomona, grippotyphosa, and autumnalis, respectively. Microscopic agglutination test titers to serovar autumnalis were higher than MAT titers to serovars pomona and grippotyphosa and persisted in some dogs for 16 weeks (6 weeks longer than for titers to serovar pomona). Of the 20 mature dogs, 13, 5, and 20 developed MAT titers of > 100 at 2 weeks to serovars pomona, grippotyphosa, and autumnalis, respectively. Titers to serovar pomona were higher and persisted in some dogs beyond 16 weeks after vaccination, compared with titers to serovars pomona and grippotyphosa, which persisted for 10 and 6 weeks, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: Subunit vaccines against serovars pomona and grippotyphosa induce MAT titers not only to homologous antigens but also to serovar autumnalis, which could lead to a misdiagnosis of leptospirosis caused by serovar autumnalis.  相似文献   
26.
A rapid, specific, and sensitive procedure for determining four widely used tetracycline antibiotics and three related epimers in bovine, swine, and poultry muscle tissues is presented. The method is based on the matrix solid-phase dispersion technique with heated water as the extractant followed by liquid chromatography (LC)-tandem mass spectrometry (MS) equipped with an electrospray ion source. Target compounds were extracted from tissues with 5 mL of water heated at 70 degrees C. After acidification and filtration, 100 microL of the aqueous extract was injected in the LC column. MS data acquisition was performed in the multireaction monitoring mode, selecting two precursor ion to product ion transitions for each target compound. Heated water appeared to be an excellent extractant, since the absolute recovery data ranged between 70 and 78%. The accuracy of the method was determined at three spike levels, using minocycline as a surrogate analyte, in any different kind of muscle tissues considered and varied between 88 and 109% with relative standard deviations ranging between 3 and 11%. Limits of quantification were estimated to range between 1 (chlortetracycline) and 9 ng/g (4-epioxytetracycline), based on a signal-to-noise ratio of 10, and are well below the tolerance levels set by the European Union. The effects of the extraction temperature, volume of the extractant, and washing of the material supporting the biological matrix with ethylenediamine tetraacetic disodium salt on the analyte recovery were studied.  相似文献   
27.
Several sulfonamide antimicrobials (SAAs) are largely used in veterinary medicine. A rapid, specific, and sensitive procedure for determining 12 SAAs in cheese is presented. The method is based on the matrix solid-phase dispersion technique followed by liquid chromatography (LC)-tandem mass spectrometry (MS) equipped with an electrospray ion source. Target compounds were extracted from Mozzarella, Asiago, Parmigiano, Emmenthal, and Camembert cheese samples by 6 mL of water modified with 10% methanol and heated at 120 degrees C. The addition of methanol to hot water served to improve remarkably extraction yields of the most lipophilic SAAs, that is, sulfadimethoxine and sulfaquinoxaline. After acidification and filtration, 100 microL of the aqueous extract was injected in the LC column. MS data acquisition was performed in the multireaction monitoring mode, selecting two precursor-to-product ion transitions for each target compound. Methanol-modified hot water appeared to be an efficient extractant, because absolute recovery ranged between 67 and 88%. Using sulfamoxole as surrogate analyte, recovery of the 12 analytes spiked in the five types of cheese considered at the 50 ng/g level ranged between 75 and 105% with RSD not higher than 11%. Statistical analysis of the mean recovery data showed that the extraction efficiency was not affected by the type of cheese analyzed. This result indicates this method could be applied to other cheese types not considered here. The accuracy of the method was determined at three spike levels, that is, 20, 50, and 100 ng/g, and varied between 73 and 102% with relative standard deviations ranging between 4 and 12%. On the basis of a signal-to-noise ratio of 10, limits of quantification were estimated to be <1 ng/g.  相似文献   
28.
Taking into account the role played by the neuroendocrine network in affecting the early development of the immune response, the present study aims to assess neonatal immunity in piglets by testing peripheral lymphocyte age-related changes in relationship to plasma levels of some relevant immunoregulatory hormones, such as growth hormone (GH), prolactin (PRL) and cortisol. For this purpose, we studied the peripheral lymphocyte age-related changes in relationship to plasma levels of GH, PRL and cortisol in conventional piglets from birth (day 0) to 41 days of age. A significant decrease was observed in the total number of lymphocytes at day 0, with a subsequent constant increment up to 41 days of age. Concomitantly, the number of T cell subsets (mainly CD8(+) cells and double positive CD4(+)CD8(+)) was low at birth, with strong increments between the 19th and 41st days of life. The CD4(+) T cell number subset was less diminished at birth than that of CD8(+), albeit with significant increments in the post-weaning period. Of interest, gammadelta T cells, which are more involved in innate immune efficiency, displayed the same trend as CD8(+) T cells from birth to the 41st day of life. From day 0 up to the 19th day, significant inverse correlations were found between T cell subsets and GH or PRL or cortisol, albeit with more significant inverse correlations with cortisol. The high levels of GH and PRL in the pre-weaning period may be due to the fact that they have to counteract the cortisol-mediated negative effect on lymphocyte production and development. These findings suggest that stress condition occurs at birth with decreases in the immune parameters, in the same way as in human newborns, with a subsequent gradual normalisation and immune development, as shown by decreased cortisol, GH and PRL normalisation and concomitant increments in T cell subsets.  相似文献   
29.
High-fidelity human patient simulators have been used for decades in medical education to provide opportunities for students to practice technical skills, diagnostic and therapeutic planning, and communication skills in a safe environment. A high-fidelity canine patient simulator (CPS) was developed using components from a human patient simulator and a low-fidelity foam core canine mannequin. Ninety-six veterinary students participated in cardiopulmonary arrest scenarios in groups of three to five students. Afterwards, participants were asked to complete an anonymous online survey describing their experiences. A total of 70 students (73%) completed the survey. All of the students (100%) felt that the simulator session expanded their cardiopulmonary resuscitation (CPR) knowledge base, and 97% responded that their skills and abilities had improved. Students also expressed positive opinions about the CPS, with 89% agreeing or strongly agreeing that the CPS was realistic and 73% agreeing or strongly agreeing that the scenarios generated emotions similar to real clinical situations. Most participants (98.5%) agreed or strongly agreed that the simulator was an engaging learning experience. Students commonly commented that the simulations allowed them to practice communication and teamwork skills and were more effective than paper-based, problem-oriented learning opportunities and lecture. Students also commented that they wanted more opportunities to participate in simulation exercises. These results suggest that high-fidelity veterinary simulation is an engaging educational methodology that addresses some limitations of other forms of problem-based learning. More studies are needed to quantitatively determine the effectiveness of this novel veterinary educational technology in comparison with more traditional approaches.  相似文献   
30.
Canine Malassezia dermatitis is frequently treated with systemic ketoconazole (KTZ) and itraconazole (ITZ). However, no information is available on the antifungal susceptibility to azoles and allilamine of Malassezia pachydermatis isolates from dogs with or without skin lesions. The present study was designed to evaluate the in vitro antifungal susceptibility of M. pachydermatis strains from dogs with or without skin lesions to KTZ, ITZ, miconazole (MICO), fluconazole (FLZ), posaconazole (POS), voriconazole (VOR) and terbinafine (TER) using the Clinical and Laboratory Standards Institute reference Broth Microdilution Method (CLSI M27-A2). The association between the susceptibility to antifungal compounds and the origin of M. pachydermatis, from skin with or without lesions has been also assessed. A total of 62 M. pachydermatis strains from healthy dogs (i.e., Group A=30) or with skin lesions (i.e., Group B=32) were tested. ITZ, KTZ and POS showed the highest activity against M. pachydermatis strains, whereas MICO TER and FLZ the lowest. A higher number of Malassezia resistant strains were registered among isolates from Group B than those from Group A. This study indicates that M. pachydermatis strains were susceptible to ITZ, KTZ, and POS. However, dogs with lesions may harbour strains with low susceptibility to antifungal agents and displaying cross-resistance phenomena to azole. The antifungal therapy in Malassezia infections requires careful appraisal of choice of drugs especially in cases of unresponsiveness to antifungal treatment or recurrent infections.  相似文献   
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