Nucleic acid staining with the methyl green-pyronin method. A comparison of the use of pure dyes and commercially available dyes

We compared the staining obtained using commercially available pyronin Y samples with that obtained using pure pyronin Y in a standardized methyl green-pyronin procedure. In addition, the importance of the dye content of the anhydrous dye was investigated by varying the dye content by the addition of pure pyronin Y to one of the commercially available pyronin Y samples. We found that, for routine histological work, commercially available pyronin Y samples may produce acceptable results provided the sample can be shown by spectrophotometry to contain at least 43% pyronin Y.     

Flow cytometric classification of biopsy specimens from cervical intraepithelial neoplasia

The distribution of single-cell DNA content was investigated in biopsy specimens from the human cervix of 121 women suspected of having intraepithelial neoplasia. Comparison of the results of the histopathological examination with the ploidy level showed that all normal specimens were diploid. Thus, no false-positive results occurred. Most of the specimens classified as mild and moderate dysplasia were diploid as well. Aneuploid cell populations occurred in 78% of the lesions classified as severe dysplasia and carcinoma in situ. The ploidy level distribution permitted a natural division of the aneuploid cell populations into two groups with DNA indices either above or below 1.5. The importance of the aneuploidy in carcinogenesis is discussed.     

Characterization of ploidy level in bladder tumors and selected site specimens by flow cytometry

The histopathological grade of atypia was compared with the ploidy level in biopsy specimens from bladder tumors and six grossly normal areas (selected sites). The study included investigation of 303 specimens from 45 patients. The results demonstrated a high correlation between the grade of atypia and the occurrence of aneuploid cell populations. The selected site specimens were diploid in patients with low grade malignant tumors, while high grade malignant tumors were associated with aneuploid selected site specimens in 20% of the cases. The divergence of ploidy level in the tumor and selected site specimens found in some cases clearly demonstrated that the carcinogenic process in the bladder may lead to clonal heterogeneity. Flow cytometric analysis seems suited for mapping of the ploidy levels in different parts of the bladder. The possible importance of the additional information is discussed.     

Loss of cellular immune reactivity during acute Plasmodium falciparum malaria

Sixteen patients suffering from acute Plasmodium falciparum malaria were studied. All were residents of an area of unstable malaria-transmission in Eastern Sudan. Blood-samples were drawn at diagnosis, and 7 and 30 days later. Blood-samples from thirteen donors, drawn outside the malaria transmission season 5 months prior to the attack, were included in the study. Lymphoproliferative responsiveness to purified soluble malarial antigens and to the unrelated antigen PPD was lost during the acute phase of the disease in most donors, but was regained during convalescence, except in four donors recrudescing or reinfected by day 30. In contrast to the suppression of antigenic responses, cellular responses to phytohaemagglutinin (PHA) remained virtually unaffected. All donors showed elevated plasma-levels of soluble IL-2 receptor during the acute phase of the disease which normalized during convalescence. Five donors examined by fluorescence-activated cell sorting (FACS) showed no increase in surface expression of IL-2 receptor on peripheral lymphocytes. The data indicate that acute P. falciparum malaria causes a depletion of antigen-reactive T-cells from the peripheral circulation, probably due to homing of this cell-population to lymphoid tissues. It was also found that acute-phase plasma was suppressive to PPD-induced proliferative responses, indicating an additional suppressive mechanism operating in vivo.     

Correlation of DNA distribution and cytological differentiation of human cervical carcinomas

The DNA distribution of biopsy specimens from 46 patients suffering from cervical carcinoma was analysed by flow cytometry and compared with the cytological differentiation. According to morphological criteria the carcinomas were classified as highly differentiated, moderately differentiated and poorly differentiated. The results demonstrate that highly differentiated tumours contain hyperploid cells predominantly with hyperdiploid DNA content. Hyperploid cell populations in the moderately differentiated tumours are mainly in the hyperdiploid and tetraploid regions. Poorly differentiated tumours contain hypertetraploid and aneuploid cell populations. The significance of these findings is discussed.     

Changes in mammary blood flow during the lactation period in goats measured by the ultrasound Doppler principle

1. Mammary blood flow was measured during the lactation period in two low- and two high-yielding dairy goats (peak milk yields approx. 1.5 and 3.6 kg/day respectively), using the ultrasound Doppler principle for determination of blood velocity in both milk veins (subcutaneous abdominal veins) of the animals, and ultrasound scanning for measurement of cross-sectional area of the veins. 2. Milk vein cross-sectional area ranged from 0.11 to 0.38 cm2 in the four goats, with a close to significant (P = 0.06) difference between the veins in the two sides of the animals. Cross-sectional area remained constant during the lactation period. Changes in mammary blood flow was therefore caused by changes in blood velocity. 3. Milk vein blood velocities ranged from 4.4 to 34.7 cm/sec independently of the time of the day, and were of the same magnitude in the two sides of the animals. Except in one goat (P = 0.1), blood velocity decreased significantly (P less than 0.01) with progressing lactation, during which period also milk yield was declining. 4. In the two low-yielding goats, a positive linear relationship (R2 = 0.20) was found between milk yield and milk vein blood velocity, whereas a diminishing exponential relationship (R2 = 0.97) was found in the two high-yielding goats. At a given milk vein blood velocity, high-yielding goats obtained a higher milk yield and also responded to changes in blood velocity (up to approx. 15 cm/sec) with greater increases in milk yield than low-yielding goats.(ABSTRACT TRUNCATED AT 250 WORDS)