Quantitative cytology of the positive region in flow sorted vaginal smears.

Fifty-one gynecologic specimens were collected from three women's hospitals and mailed in a prefixed status to our laboratory. The specimens were classified into a negative, a suspicious, a postradiation, and a positive group. After single cell dispersion the samples were stained for DNA and protein, analyzed, and sorted in the dual laser equipped Heidelberg flow analyzer sorter (HEIFAS). Particles with elevated DNA values (beyond 3.5 ploidy) and with intermediate protein values were sorted as the positive fraction directly on microscopic slides. After restaining according to Papanicolaou, they were re-evaluated cytologically and identified as tumor cells, dysplastic cells and false alarms. The latter consist of doublets and aggregates of more than two cells, binucleated cells, sperm aggregates and epithelial cells contaminated with bacteria. The different groups showed significant differences regarding the total rate of aggregates to single cells. In general, false alarms were very frequent in the positive region and impeded the statistical classification of the sample. The reduction of false alarms is a prerequisite for prescreening with flow instrumentation.     

Cytobrush and wooden spatula for oral exfoliative cytology. A comparison.

The Cytobrush has been used frequently in cervical cytology, but as yet its value in oral exfoliative cytology has not been assessed. A study was undertaken to compare the efficiency of the Cytobrush with that of the wooden tongue spatula. For 26 patients, two smears were collected from clinically normal mucosa from four sites in the oral cavity (buccal mucosa, dorsal tongue, ventral tongue and hard palate). The smears were graded for cell yield and dispersion on a three-point scale. The results were analyzed using the chi 2 test. The Cytobrush was found to be significantly more efficient than the wooden spatula, in terms of both cell yield (P less than .005) and cell dispersion (P less than .005). When each site was examined separately, the Cytobrush produced significantly better dispersion for the dorsal tongue, ventral tongue and buccal mucosa and a better cell yield for the tongue surfaces. No significant difference for cell yield or dispersion was found for the hard palate. The study showed that the Cytobrush is an effective instrument for use in exfoliative cytology of normal oral mucosa.     

The measurement of human endometrial prostaglandin production. A comparison of two in vitro methods

Two in vitro methods for measuring human endometrial prostaglandin production were compared. Endometrial samples from eight patients were incubated over eight hours by a perifusion and a superfusion technique. The collected fractions were assayed by radioimmunoassay for PGE2 and PGF2 alpha. There was no significant difference between the perifusion and superfusion methods for the pattern and amount of PGE2 and PGF2 alpha production with time. Significantly higher production levels of PGE2 and PGF2 alpha were found in secretory phase endometria than in proliferative phase endometria. Histological examination of the tissue specimens by light and electron microscopy showed that both methods caused gross tissue damage after eight hours experimentation. The superfusion method produced more morphological damage than the perifusion method. However, no tissue damage could be detected after one hour of incubation with either method. Over an eight hour period neither the perifusion nor the superfusion technique appears to be a good indicator of in vivo endometrial prostaglandin production. Either technique used for only one to two hours may better reflect the in vivo situation.     

Liquid-based cytology and conventional smears compared over two 12-month periods.

BACKGROUND AND OBJECTIVE: Liquid based cytology (LBC) was introduced across the Scottish Cervical Screening Programme in 2003-2004. The objective of this study was to compare in a large cytopathology laboratory the results of cervical samples over two twelve-month periods, 2001-2002, when the great majority of smears were conventional, with 2003-2004, when all were LBC. METHODS: The results of smears in both periods were analysed to give overall reporting profiles, and correlated with results of cervical biopsies. The numbers of patients referred for colposcopy were compared. RESULTS: The percentage of unsatisfactory smears fell from 13.6% to 1.9%. Colposcopic referrals for repeated unsatisfactory smears fell from almost 25% to 0.5%. There was a decrease in overall smear numbers, but despite this there was an increase in the number of smears reported as showing dyskaryosis of any grade. There was an increase in positive predictive value for moderate dyskaryosis and above, from 79.5% to 86.1%. The outcome of biopsies from patients referred with mild dyskaryosis showed no decrease in accuracy of predicting a low grade histological lesion. Workload in the laboratory decreased, due to fewer smears received overall, more rapid primary screening times and fewer multi-slide cases. Primary screening backlogs all but disappeared, and reporting times greatly improved. CONCLUSIONS: Introduction of liquid based cytology led to improvements in unsatisfactory smear rates, with significant benefits to colposcopic referrals and laboratory turnaround times. Pick-up rates of dyskaryosis were maintained, and the positive predictive value of a dyskaryotic smear report was improved.     

A multi-spectral approach for scene analysis of cervical cytology smears.

A multi-spectral approach for the scene analysis of cervical cytology smears, using multiple images of a scene digitized through suitably chosen color filters matched to the Papanicolaou stain, has been proposed here. This technique involves clustering of two-dimensional data for extracting cytoplasm of the epithelial cells. Its performance on an experimental data set of 233 scenes involving more than 10 types of normal and malignant epithelial cells has been compared with density and gradient thresholding techniques. This resulted in an approximate 83% rate of success compared to approximately 40% for the rest of the other techniques.     

Quantitative cytomorphologic analyses of Papanicolaou-stained smears from oral lichen planus

OBJECTIVE: To apply quantitative techniques to cytologic smears in order to improve the diagnostic sensitivity of cytology in the detection of malignant change in the oral cavity. STUDY DESIGN: Eighty-two patients with lesions of oral lichen planus were investigated. A total of 247 Papanicolaou-stained buccal smears underwent nuclear and cytoplasmic area measurements using a TV image analyzer. RESULTS: The cytomorphologic results of this study suggest that lesions of oral lichen planus contain smaller cells and nuclei than those observed in clinically normal oral mucosal smears. There was a statistically significant reduction in nuclear area (P < .001) and cytoplasmic area (P < .05) for the oral lichen planus smears. CONCLUSION: The TV image analysis system could provide a valuable method of quantifying cell changes in oral smears collected from oral mucosa as a method of monitoring lesions of oral lichen planus.     

Quantitative oral exfoliative cytology. Effect of alcohol on normal buccal mucosa

OBJECTIVE: To assess the effect of chronic alcohol intake on the DNA distribution and cell area of normal oral mucosal cells. STUDY DESIGN: Smears were taken from clinically normal buccal mucosa of 50 patients attending an alcohol-problem service (i.e., chronic alcohol use) and average alcohol units per week recorded. DNA distribution histograms and total cell area values were then compared to those obtained from smears taken from a control group (which included social drinkers) of patients attending for routine dental treatment. Nuclear DNA content was assessed on 100 randomly selected, Feulgen-stained nuclei using a Seescan TV image analysis system, and total cell area was assessed on 50 Papanicolaou-stained cells using the Vids V image analysis system. RESULTS: The DNA distribution histograms were essentially diploid in appearance for the alcohol group, although there was an increase in nuclear DNA content in the occasional nucleus. A highly significant reduction in total cell area was found for the alcohol group when compared to the controls. CONCLUSION: The chronic ingestion of alcohol is associated with a reduction in total cell area but appears to have little effect on nuclear DNA content. Our previous research using the same technique showed that oral cancers are frequently nondiploid. Thus, a nondiploid DNA distribution histogram for smears taken from a clinically suspicious lesion in someone who consumes excessive amounts of alcohol is unlikely to be due to alcohol use alone and should indicate biopsy.     

Quantitative exfoliative oral cytology in iron-deficiency and megaloblastic anemia

A study was undertaken to ascertain if there were any morphometric or morphologic changes in exfoliated oral squames in either iron-deficiency or vitamin B12-deficiency states. The results revealed a significant (P less than .05) increase in nuclear area and a significant alteration in nuclear/cytoplasmic ratio in vitamin B12 deficiency; both returned to normal following replacement therapy. No changes were seen with iron deficiency anemia or non-vitamin B12 megaloblastic anemia. Ultrastructurally, the surface morphology showed similar features in all groups, with the plasma membrane forming complex folds (microplications) in three patterns: branching, parallel and network. The microplication widths and interplication distances were remarkably constant in both control and study groups, regardless of pattern.     

A comparison of methods for the measurement of protein turnover in vivo.

Steady-state rates of turnover of two single proteins were measured in vivo by two independent methods. The fractional rate of synthesis of liver ornithine aminotransferase, measured by a continuous infusion of L-[2,6-3H]tyrosine, was 0.42 day-1, whereas in the same animals the fractional rate of degradation measured by loss of radioactivity from amino acids labelled via [14C]bicarbonate was 0.40 day-1. The agreement between methods confirms the reliability of each method for the study of hepatic protein turnover. In contrast, [14C]bicarbonate-labelled amino acids are extensively reutilized in muscle, and are therefore unsuitable for measuring rates of muscle protein breakdown.     

Standardization of fine needle aspiration cytology of the breast -- comparison of Auto Cyto Fix and conventional smears.

In Japan, there are some problems with fine needle aspiration (FNA) cytology of the breast, such as insufficient smeared cells, air-drying artefact and excessive erythrocytes. Liquid-based cytology has been found to solve these problems. Equipment for such preparations has been developed, but can be expensive to purchase and operate. We developed Auto Cyto Fix 1000 (ACF), which is inexpensive and automatically smears and fixes cells. The purpose of this study was to compare the various cytological features of conventional and ACF specimens. We evaluated whether the ACF method would be able to replace the conventional method. Forty-eight FNA specimens of breast were studied. All specimens were prepared by the direct smeared (DS) and ACF methods and evaluated for unsatisfactory cell collection, air-drying artefacts, background findings and epithelial cell findings. Although ACF specimens were prepared using the cells remaining in the needle and syringe after preparing DS specimens, the cellularity of two of the ACF specimens was better than that of the corresponding DS specimens. ACF specimens never showed air-drying artefact. Unlike DS specimens, which have many erythrocytes in the background, erythrocytes were filtered out and the background of ACF specimens was clean. We believe that many problems attributable to conventional FNA specimen preparation have been solved in this study. Preparation using the ACF apparatus can reduce running costs and can be used to prepare FNA specimens of the breast for cytological examination as an alternative to the conventional method.     

Quantitative analysis of flavonol glycosides in Ginkgo biloba: a comparison of two analytical methods

A comparison of testing methods for constituents of Ginkgo biloba was performed as part of the process for determining which of the many available analytical methods was most appropriate for inclusion in the American Herbal Pharmacopoeia (AHP). Three primary methods were reviewed: those adopted by the European Pharmacopoeia, the National Science Foundation-Institute for Nutraceutical Advancement (NSF-INA) and the United States Pharmacopeia. Methods were compared by evaluating sample preparation and hydrolysis, precision and methods of total flavonol glycoside calculation by two independent laboratories. The AHP adopted the method of NSF-INA for inclusion in the AHP monograph owing to observed superiority of this method in terms of precision, glycoside calculation, ease of sample preparation and chromatographic parameters.     

Esophageal exfoliative cytology samplers. A comparison of three types

OBJECTIVE: To compare three types of esophageal exfoliative cytology samplers in terms of patient acceptability, ease of use, final destination of the sampler tip in the gastrointestinal tract and cellular yield. STUDY DESIGN: A controlled, single-blind, cross-over study was undertaken to compare the balloon, sponge and sponge-mesh samplers in healthy volunteers. After completing the three procedures in random order, participants were asked about their preferred method. Ease of use was defined as the technician's ability to perform the intubation successfully. Final destination of the samplers was assessed fluoroscopically. Cytopathologists determined the cellular yield of each sampler using the Bethesda System. RESULTS: Sixty-two volunteers participated. The two encapsulated samplers were significantly preferred over the balloon (P < .0001). There was no significant difference in ease of use, final destination or cellular yield of the three techniques. All three samplers were successfully intubated on the first attempt and retrieved adequate numbers of squamous and glandular cells in > 78% of cases. CONCLUSION: All three samplers obtained satisfactory yields of squamous and glandular cells, but the encapsulated samplers were more patient acceptable. The sponge-mesh sampler may be the least complicated sampler for field screening use. Larger-scale studies will be required to test the accuracy of these three samplers for detecting esophageal dysplasia and carcinoma.     

A comparison of two autoradiographic methods for detecting radiolabeled nucleic acids in embryos.

Two methods for preparing embryos for autoradiographic study of newly synthesized nucleic acids are described and compared. The first method consists of rapidly fixing radiolabeled embryos with acetic acid:methanol, spreading them on glass slides and exposing them for 8 days with a photographic emulsion. The second method consists of fixing, embedding in resin, and sectioning the embryos before their exposure with the emulsion for 3 weeks. Both techniques have many applications in studies of early embryonic activity, but the spread technique is very sensitive, simpler, and faster.     

Vaginal and oral cytology of the menopause. A comparative study

A comparative cytologic study was made of the hormonal content of the vaginal and oral mucosa of menopausal women. Twenty-three women between one and ten years (early menopause) and 33 patients more than ten years (late menopause) after cessation of menstruation had oral and vaginal smears taken. Comparable smears from 21 young women, and oral smears from 18 males served as controls. The smears were evaluated by the maturation value method, and averages were established for each group. The vaginal smears had low maturation values of 40 and 22 in the early and late menopausal groups, respectively. In these women, the buccal smears' maturation values were high, above 70. Similar high maturation values were found in buccal smears of the young women and men. It was concluded that the high maturation values found in the oral smears of the menopausal women are not the result of hormonal effect but that of local mechanisms or irritative factors.     

A comparison of two methods of infiltration in breast reduction surgery

The superwet technique has been shown in previous studies to dramatically reduce blood loss in breast reduction surgery, compared with standard infiltration. A retrospective chart review of 303 consecutive patients undergoing bilateral breast reduction surgery was undertaken to demonstrate additional differences in complication rate, operative time, or sponge use in the operating room. In this series, 132 consecutive patients received standard infiltration along incision lines (25 cc per breast of 1:100,000 epinephrine), and 171 patients received superwet infiltration with 240 cc per breast of 1:1,000,000 epinephrine. The average operative time was significantly reduced in the superwet group, from 78.5 minutes to 70.7 minutes (p < 0.01 level). The average number of sponges used intraoperatively was also decreased significantly (p < 0.01), from 26 to 20 sponges. Complication rates were equally low in both groups, demonstrating the safety of the superwet technique. In addition to limiting blood loss, the superwet infiltration effectively reduces operative time and sponge use without increasing complications in breast reduction surgery.