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Biomedical subjects

L Maxia

Publications and source records attributed to L Maxia.

At least 19 recordsLinked to original sources

Detection of Leishmania infantum in dogs by PCR with lymph node aspirates and blood.

The PCR technique was applied to the diagnosis of leishmaniasis in dogs, both serologically negative and positive. DNA was taken from lymph node aspirates and blood. The primers 13a and 13b, derived from Leishmania amazonies and Leishmania braziliensis kinetoplast DNA (kDNA), also amplified Leishmania infantum IPT1 constant region of minicircle kDNA. The amplified fragment is 116 bp long. It was cloned and the sequence was determined. A 70-bp inner fragment was designed and used as a probe in dot blot hybridization. A group of 124 dogs was examined, 37 of which showed typical symptoms of disease. PCR was performed on 124 blood samples and 52 lymph node aspirates. Using microscopic examination as the "gold standard," we calculated sensitivity, specificity, and positive and negative predictive values of 100% using lymph node aspirates and values of 85, 80, 95, and 57%, respectively, using blood samples. We found that 40% of the animals without lesions and 38% of the animals with clinical signs gave false-negative results by indirect immunofluorescence antibody testing. These animals could contribute to the spreading of infection among dogs, and represent a potential risk for human health as well.

Animals↗

Characterization of Theileria species by PCR using specific target sequences.

Theileriosis is an infectious disease in tropical countries and in the Mediterranean area. It is caused by Theileria, a haemoprotozoan, transmitted by vectors belonging to the Ixodidae. In Southern Italy and in Sicily the infection is due mainly to T. annulata, but in some cases other species are involved in the disease. The authors describe a method to identify theileriosis in cattle blood samples, using PCR and hybridization techniques. Different primer sets were used to amplify different DNA target sequences, both genus and species specific. Blood samples from cattle were collected in Sicily. The DNA extracted from blood samples was employed first to detect the presence of the 18S ribosomal subunit gene specific for Theileria genus. Successively the positive samples were analysed to identify the species, T. annulata or T. buffeli/orientalis, using as target sequences for amplification respectively a fragment of the TAMS-1 and p33/34 antigens gene. Here the authors describe for the first time the presence of T. buffeli/orientalis infection in Sicilian herds. In fact 66% of positive blood samples were T. buffeli/orientalis infected.

Animals↗

Detection of Mycobacterium tuberculosis complex in cattle by PCR using milk, lymph node aspirates, and nasal swabs.

The PCR technique was applied to the diagnosis of tuberculosis in live cattle, and both skin-test-negative and skin-test-positive animals were studied. DNA was taken from various sources including specimens of lymph node aspirates, milk, and nasal swabs. After slaughter and visual inspection, tissues such as lymph nodes, lungs, and udders from tuberculin reactors were tested by the same technique. Specific oligonucleotide primers internal to the IS6110 insertion element were used to amplify a 580-bp fragment. A 182-bp fragment was obtained by designating a nested PCR from the first amplification product. This fragment was cloned and sequenced, and after being labeled it was employed in dot blot hybridization. A total of 100 cattle were tested, and PCR analysis was performed using nasal swab, milk, and lymph node aspirate. Sixty skin-test-positive cows were also tested to detect mycobacterial DNA in tissue samples from lymph nodes, lungs, and udders, and the infection was confirmed in all of the animals. Using PCR analysis of tissue samples from slaughtered animals as a "gold standard" we calculated 100% values for sensitivity, specificity, and positive and negative predictive values for milk and lymph node aspirate samples. The respective values for nasal swab samples were 58, 100, 100, and 28%. The respective values for all of the samples were 74, 100, 100, and 35%, while for visual inspection the values were 81, 100, 100, and 58%, respectively. PCR analysis of specimens of lymph node aspirates, milk, and nasal swabs from skin-test-negative animals showed that 52% of these skin test results were false negatives. These animals, not being removed from the farms, represent a potential source of further infection.

Animals↗

Characterization of Langmuir-Blodgett films of rhodopsin: thermal stability studies.

Two-dimensional close packing of purified bovine rhodopsin, made by the Langmuir-Blodgett technique, was characterized by small angle x-ray scattering and nanogravimetric measurements. The area occupied by a molecule of rhodopsin in the film was approximately 1100 Angstrum2 and the periodicity of the layers resulted in 59 Angstrum. The circular dichroism measurements showed that bleached rhodopsin in Langmuir-Blodgett film had high thermal stability, in fact, reaching a temperature of 150 degrees C without a loss of the secondary structure. Moreover, when the film was made up in the dark, rhodopsin maintained its stability up to at least 200 degrees C and its characteristic absorbance peak at 500 nm up to about 90 degrees C.

Animals↗