This study established a modified alkaline phosphatase-labelled avidin-biotin-complex (ABC-AP) method for diagnosis of mouse hepatitis virus (MHV) and Mycoplasma pulmonis infection from formalin-fixed, paraffin wax-embedded sections, murine antibody-positive serum being used as the primary reagent. With this method, MHV antigen in cAnNCrj.Cg-Foxn1(nu)/Foxn1(nu) mice and M. pulmonis antigen in Wistar rats were immunolabelled in tissue sections. MHV antigen was clearly detected in samples of liver, stomach, caecal and colonic mucosa, and spleen. M. pulmonis antigen was demonstrated on the luminal surface of bronchiolar epithelial cells. This method may prove useful in diagnosis when commercial antisera are unavailable or when immunosuppression prevents serological diagnosis.

Animals↗

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Reference recombinant chinese hamster ovary cell lines.

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Animals↗

PubMedDate not supplied

Determination of daminozide in apples by gas chromatography/chemical ionization mass spectrometry.

A method using gas chromatography/chemical ionization mass spectrometry (GC/CIMS) for the determination of daminozide residues in apples has been developed. Daminozide was separated from the sample matrix by water extraction and cation exchange, converted to the methyl ester by treatment with HCl-methanol, and determined by GC/CIMS using succinonitrile as an internal standard. The detection level was 0.05 ppm. Recoveries were 92-104% from apples spiked at the 0.05-0.5 ppm levels. Of the 25 apple samples analyzed, only 2 were positive for daminozide (1.04 and 0.32 ppm).

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Treatment of refractory or relapsed adult acute leukemia by using mitoxantrone-containing regimens.

Twenty-eight adult patients with primary refractory or relapsed acute leukemia were treated. The regimens consisted of mitoxantrone plus cytosine arabinoside for 17 patients with acute non-lymphocytic leukemia (ANLL) and mitoxantrone accompanied with vincristine and prednisolone for 11 patients with acute lymphoblastic leukemia (ALL). In primary refractory patients, 1 of the 4 (25%) ANLL and 1 of the 3 (33%) ALL attained complete remission (CR). Excluding 2 patients who underwent bone marrow transplantation, 8 of the 13 (62%) relapsed ANLL and 4 of the 8 (50%) relapsed ALL achieved CR with a median duration of remission of 6.2 months and 3.8 months, respectively. Myelosuppression occurred in all treatment courses and was associated with pyrexia due to infections in 84% of the cases. Nausea, vomiting and stomatitis were mild. Abnormal liver function tests were observed in 8 (28%) patients. One patient, pretreated with 550 mg/m2 of doxorubicin, developed congestive heart failure. The results suggest that mitoxantrone is of value in the treatment of Chinese patients with refractory or relapsed acute leukemia.

Adult↗

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Fragile-X mental retardation--a combination of cytogenetic and molecular approaches, with greater emphasis on DNA analysis.

Fragile X syndrome is one of the most frequent causes of hereditary mental retardation. In the past, its diagnosis depended primarily on cytogenetic demonstration of chromosome fragile site Xq27.3. Recently, the gene FMR-1 has been found responsible for this disease. Here a combined method was used to study fragile X syndrome. A fragment (pP1fr) of DNA was subcloned from pE5.1 by polymerase chain reaction. With this probe, DNA samples from two cytogenetically proved families were analyzed by restriction fragment length polymorphisms. It was demonstrated that EcoRI polymorphism was an easy and accurate method for diagnosis of the fragile X syndrome. To study methylation status of patients, another methylation-sensitive enzyme, BssHII, could be used together with EcoRI. The PstI polymorphism of one family was also studied and showed one kb fragment as normal, and detected more precise changes in length. Prominent mosaicism necessary was characteristic in PstI polymorphism. The DNA diagnosis of fragile X syndrome was a reliable method.

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PubMedDate not supplied

Y-specific polymerase chain reaction for the interpretation of a chromosome marker.

Chromosome marker is a very special type of chromosomal aberration. In addition to their normal chromosomes, patients with this type of disease own one (or more) small, unrecognizable chromosome. The extra chromosome markers may originate from the products of breakage and translocation of other chromosomes. Interpretation of the chromosome marker is usually very difficult, but its presence is an important prognosis factor for patients if the marker originated from a Y chromosome, or its components. This study explores the possibility of interpreting the chromosome marker by Y-specific polymerase chain reaction. A pair of primers designed from one Y-specific DNA sequence was employed. By studying normal controls, the most specific conditions for the chain reaction were established. Comparison of normal males and females showed that this reaction is specific to males (that is, to the Y chromosome). Also one patient with a karyotype 46,X,+marker was studied. The external genitalia of this patient were female type except for an enlarged clitoris. Y-specific polymerase chain reaction showed that the patient did have the Y-specific sequence. The chromosome marker did originate from the Y chromosome. This work proposes that Y-specific polymerase chain reaction is an effective and necessary diagnostic procedure for patients with chromosome marker.

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