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PubMed2006

Morphometric assessment of the impact of serum thymulin immunoneutralization on pituitary cell populations in peripubertal mice.

Thymulin is a thymic hormone involved in several aspects of intra- and extrathymic T-cell differentiation. Thymulin also possesses hypophysiotropic activity which suggests that this metallopeptide may play an important role in thymus-pituitary communication, particularly during early life. The aim of the present study was to evaluate the impact of serum thymulin suppression from birth to peripuberty on the morphology of different pituitary cell populations in prepubertal C57Bl/6 mice. Animals were submitted to immunoneutralization of circulating thymulin from postnatal day 1 to the end of the study (age 32 days). From their 1st day of life, the animals were submitted to a protocol of intraperitoneal injections of rabbit anti-thymulin serum (alpha-FTS) and normal rabbit serum (NRS) in the controls. On their 33rd day of life, the animals were killed and their pituitaries were immediately dissected, fixed and immunostained using the EnVision system with primary antibodies against growth hormone, thyrotropin, corticotropin, gonadotropins and prolactin. Morphometry was performed by means of an image analysis system. The following parameters were calculated: volume density = Sigma cell area/reference area (RA); cell density (CD) = number of cells/RA, and cell size (expressed in microm2). Serum thymulin was measured by a rosette bioassay while pituitary hormones were assayed by radioimmunoassay. Serum prolactin, luteinizing hormone, follicle-stimulating hormone, growth hormone and thyroid-stimulating hormone were significantly lower in the alpha-FTS animals of either sex compared with the corresponding NRS counterparts. The somatotrope, lactotrope and corticotrope populations showed a significant decrease in CD, while cell hypertrophy was observed in some of the pituitary cell populations of the alpha-FTS group compared to the NRS group. In the alpha-FTS group, there were sex differences in the morphometric changes observed. Our results suggest that serum thymulin plays a significant role during early life in the postnatal maturation of endocrine cells of the mouse anterior pituitary gland.

Adrenocorticotropic Hormone↗
PubMed1995

Adaptations of diaphragm neuromuscular junction following inactivity.

We hypothesized that differences exist in the morphological adaptations of neuromuscular junctions (NMJs) on different fiber types in response to prolonged inactivation. Two weeks of inactivity of both phrenic motoneurons and diaphragm muscle was induced by spinal cord hemitransection at C2 (spinal isolation; SI). A three-color fluorescent immunocytochemical technique, combined with laser-scanning confocal microscopy, was used to create two- (2D) and three-dimensional (3D) images of NMJs and obtain morphological information concerning: (1) innervating axons and presynaptic nerve terminals; (2) motor endplates (postsynaptic apparatus consisting of acetylcholine receptors), and (3) myosin heavy chain (MHC) phenotype of muscle fibers. In both sham controls (CTL) and SI animals, planar (2D) and surface (3D) areas of motor endplates and nerve terminals on type II muscle fibers (anti-fast MHC immunoreactive) were smaller than on type I (nonimmunoreactive to anti-fast MHC) fibers, when normalized for fiber diameter. The number of branches, total branch length and perimeter of both motor endplates and nerve terminals were greater for NMJs on type II fibers than on type I fibers. The extent of overlap between nerve terminal and endplate was greater on type I fibers than on type II fibers. After SI, there was a significant expansion of NMJs on type II fibers. Planar and surface areas of motor endplates and nerve terminals, number of endplate and nerve terminal branches, total branch length, and perimeter were all increased on type II fibers following SI. The extent of overlap of nerve terminal and endplate increased on type II fibers, approaching that observed in type I fiber NMJs. These results indicate that neuromuscular inactivation leads to a selective expansion of type II fiber NMJs through addition of new terminal area, and elongation of existing terminal branches. These changes may represent a compensatory effort to improve neuromuscular transmission.

Adaptation, Physiological↗
PubMed1997

Characterization of the nuclear deoxyribonucleic acid content and nuclear morphometry in 71 primary cutaneous melanomas.

BACKGROUND: While the determination of nuclear deoxyribonucleic acid (DNA) content (DNA ploidy level) and nuclear morphometry characterization has proved to be of prognostic value in melanocytic lesions, there are several ways of performing these determinations. OBJECTIVE: To identify which of 9 DNA ploidy- and 2 nuclear morphometry-related variables are of prognostic and/or diagnostic value in 71 primary melanomas. METHODS: Histological typing, Breslow depth determination, the evaluation of Clark's level of invasion and the 11 quantitative variables (calculated in Feulgen-stained nuclei using computer-assisted microscope analysis) determined for each melanoma were submitted to discriminant analysis. RESULTS: The discriminant analysis of image cytometric variables enabled specific cell subpopulations to be identified in histological and the Breslow-related groups, but not in the Clark-related ones. CONCLUSION: The characterization of melanoma heterogeneity by means of the identification of specific DNA ploidy level-related cell subpopulations in specific Breslow-related groups enables the problem of intra- and interobserver variability in Breslow depth determination to be reduced and therefore can help dermatologists in their daily routine.

Adult↗
PubMed1996

Comparison of ureteropelvic transitional cell carcinoma with bladder transitional cell carcinoma using an image analyzer.

To elucidate whether a significant difference in malignant potential between ureteropelvic transitional cell carcinoma (TCC) and bladder TCC is present, the argyrophilic nucleolar organizer region (AgNOR), nuclear DNA content and mean nuclear area (MNA) were evaluated in 32 ureteropelvic TCCs and 60 bladder TCCs, and were also compared with histological grade and stage. The parameters used were the mean AgNOR count (C-AgNOR), the percentage of cells exhibiting more than 3 AgNOR dots within nuclei (P-AgNOR), 2c deviation index (2cDI), 5c exceeding rate (5cER) and MNA. No significant difference in these 5 parameters was found between ureteropelvic and bladder TCCs. In addition, within each histological grade of tumor, no significant difference in each parameter between ureteropelvic and bladder TCCs was noted. Similarly, within pTa or pT1 TCCs, there was no significant difference in the parameters between ureteropelvic and bladder TCCs. On the other hand, invasive TCCs (pT2 and higher), the C-AgNOR and P-AgNOR were significantly higher in ureteropelvic than in bladder TCC (p < 0.05), while no significant difference in 2cDI, 5cFR and MNA was noted. These findings suggest that: (i) as a whole, the difference in malignant potential between ureteropelvic and bladder TCCs appears to be less significant, and (ii) factors other than the malignant potential of a tumor might contribute to the unfavorable clinical outcome in patients with ureteropelvic TCC.

Aged↗
PubMed1996

Cytofluorometric evaluation of nuclear DNA content distribution in renal neoplasms treated by conservative surgery.

The prognostic value of nuclear DNA content in 21 patients who underwent conservative surgery for renal tumors was the object of a retrospective study. Cytofluorometric evaluation of nuclear DNA content distribution was performed on smears obtained from 50-micron thick histologic sections prepared with the Hedley technique, using a Leitz MPVII microspectrophotometer. The DNA indexes were plotted in the form of frequency histograms. DNA measurements were repeated by a cell image analysis system (CAS 200; Becton Dickinson) in 16/21 cases. Nuclear DNA content was diploid in 12 cases, triploid in 4, tetraploid in 2 and multiclonal in 3. No statistically significant correlation was found between ploidy and tumor stage and size, using Kendall's tau test, while there was a significant correlation between tumor ploidy and nuclear grade (p < 0.01). Excluding 2 postoperative deaths, with an average follow-up of 54 months, tumor diffusion was observed in 2 patients (1 with multiclonal and 1 with triploid DNA content) and a local recurrence in a patient with a triploid tumor. Of the remaining 16 no evidence of disease (NED) cases, 10 have a diploid DNA tumor content, 2 diploid, 2 tetraploid and 2 multiclonal. It is concluded that even after conservative surgery for renal neoplasms a diploid DNA content is a favorable prognostic factor. A completely negative prognostic impact of nondiploid tumors is not confirmed, since although all three negative events occurred in this group, there are also 4 NED patients with respectively 70, 46, 43 and 40 months' follow-up.

Adult↗
PubMedDate not supplied

Implementation and evaluation of a new automated interactive image analysis system.

OBJECTIVE: To compare automated interactive screening using the ThinPrep Imaging System with independent manual primary screening of 12,000 routine ThinPrep slides. STUDY DESIGN: With the first 6,000 cases, the Review Scopes (RS) screening results from the 22 fields of view (FOV) only were compared to independent manual primary screening. In the next 6,000 cases, any abnormality detected in the 22 FOV resulted in full manual screening on the cytotechnologist's own microscope. Sensitivity and specificity together with their 95% CIs were calculatedfor each method. RESULTS: In the first set of 6, 000 cases, diagnostic sensitivity and specificity of the imager were 85.19% and 96.67%, respectively. The diagnostic sensitivity and specificity of manual primary screening were 89.38% and 98.42%. This highersensitivity and specificity of manual primary screening were found to be statistically significant. The second set of 6,000 cases demonstrated no significant statistical difference in sensitivity or specificity between the sets of data. CONCLUSION: The results from our study show that the sensitivity and specificity of the imager technology are equivalent to those of manual primary screening. The system is ideally suited to the rapid screening of negative cases, allowing increased laboratory productivity and greater throughput of cases on a daily basis.

Carcinoma↗
PubMedDate not supplied

Use of mailed urine specimens in diagnosing urothelial carcinoma by cytology and DNA image analysis.

OBJECTIVE: To evaluate the usefulness of urine specimens collected via a mailer system and analyzed by cytology and DNA ploidy for the detection of urothelial carcinoma (UC). STUDY DESIGN: We retrospectively reviewed the diagnoses of 91 mailed urine specimens received from 72 patients, 67% of whom had a history of UC. The specimens were fixed in an equal volume of 50% ethanol solution before being mailed. The cytologic findings were interpreted in conjunction with DNA ploidy image analysis. We compared these initial diagnoses with those of follow-up examinations, including biopsies, cystoscopic findings and urinary cytology/DNA ploidy analyses. In addition, to examine the quality of the mailed samples, 3 cytopathologists performed a blinded assessment of cytologic slides of 20 mailed and 17 fresh urinary samples for bacterial overgrowth, urothelial degeneration, and presence of proteinaceous material and crystals. RESULTS: Follow-up was available for 68 of the 91 mailed specimens. The sensitivity for detecting UC using mailed urine specimens that were analyzed by both cytology and DNA ploidy was 61%, while specificity was 92%. The levels of bacterial overgrowth and urothelial degeneration in the mailed specimens were not significantly greater than in the fresh specimens (p>0.05). The levels of proteinaceous material and crystals were significantly higher in the mailed specimens (p<0.05). CONCLUSION: The results of combined cytology and DNA ploidy image analysis by using mailed urine samples were comparable to those of fresh urine specimens for the detection of UC reported in previous publications. The increase in crystals and proteinaceous material did not impede diagnostic interpretation. The mailing system is a reliable and convenient method of monitoring and triaging patients with UC or related symptoms.

Adult↗
PubMedDate not supplied

Grading follicular lymphoma on fine needle aspiration specimens. Comparison with proliferative index by DNA image analysis and Ki-67 labeling index.

OBJECTIVE: To determine whether follicular lymphoma (FL) can be graded on fine needle aspiration (FNA) biopsies by determining the percentage of centroblasts in the neoplastic follicles on the smears. STUDY DESIGN: Eighty-nine cases of histologically confirmed cases of FL, including 31 grade 1, 46 grade 2 and 12 grade 3, were evaluated. Proliferative index (PI) by DNA image analysis (DIA) and Ki-67 labeling index (LI) were obtained on all cases. A minimum of 200 cells were counted per case (range, 200-800 cells) at 40x magnification, and the number of large cells (centroblasts) was expressed as a percentage of the total number of cells counted within the follicles. RESULTS: The percentage of centroblasts in the follicular aggregates was 9.7 +/- 2.9% in grade 1 FLs, 24.7 +/- 5.6% in grade 2 and 48.4 +/- 7.5% in grade 3. These differences were significant (P < .05). DNA image analysis of PI and Ki-67 LI differed significantly between grade 1 FLs and grade 2 and 3 FLs (P < .05), but there were no significant differences between grade 2 and 3 FLs. CONCLUSION: Determining the percentage of centroblasts in the follicular aggregates on FNA specimens is a good method of grading FLs. Using the percentage of centroblasts per follicular structure, FL grades 1, 2 and 3 were adequately distinguished. PI by DIA and Ki-67 LI clearly distinguished FL grade 1 from FL grades 2 and 3; however, it did not clearly distinguish between grades 2 and 3.

Adult↗
PubMedDate not supplied

Tumor markers in the detection of recurrent transitional cell carcinoma of the bladder: a brief review.

OBJECTIVE: To determine the status of the most promising tumor markers of bladder cancer, including comparison with cytology, technical complexity and utility in patient management. STUDY DESIGN: An extensive literature search was performed, and multiple markers were evaluated. The markers with the greatest potential for use as an adjunct to cytology were reviewed to determine the value of clinical implementation. Markers with a paucity of clinical research and poor results in clinical trials were omitted from review, as were genetic and cytologic prognostic determinants. RESULTS: NMP22, bladder tumor antigen, fibrin/fibrinogen degradation products, telomerase and QUANTICYT image analysis cytometry produced the most favorable and reproducible results. Each test obtained favorable sensitivities in comparison with cytology, especially in the detection of low grade lesions. Many also retrospectively placed patients in high- and low-risk groups based on the test results, allowing increased follow-up time between cystoscopies. However, inability to detect some high grade lesions reduces their utility. CONCLUSION: Continued clinical trials using these and other predictors of bladder cancer will eventually find a test that is suitable, in sensitivity and specificity, for use in urology clinics. Until that time, these tests may be useful in conjunction with cytology to prolong the interval between cystoscopies.

Biomarkers, Tumor↗
PubMedDate not supplied

Adrenal cortical carcinoma with a liver metastasis in a child. Report of a case with fine needle aspiration findings and image analysis of nuclear DNA.

BACKGROUND: Adrenal cortical carcinoma is a rare and highly malignant neoplasm. Liver metastases from it may require special differential techniques in addition to cytologic findings. CASE: A 14-month-old child had adrenal cortical carcinoma and a liver metastasis confirmed by fine needle aspiration biopsy (FNAB). Histologic and clinical criteria of malignancy only partially applied to this tumor. Immunohistochemical analysis of both the primary tumor and metastasis supported an adrenal cortical origin. Other ancillary tests-DNA ploidy, proliferation index (proliferating cell nuclear antigen, Ki-67) and p53 protein immunolocalization-were utilized in a diagnostic sequence. Flow cytometric and image analysis of DNA ploidy of the primary tumor gave similar results. Assessment of DNA content of the adrenal cortical carcinoma and liver metastasis by image analysis showed identical patterns. CONCLUSION: The applicability of new diagnostic techniques to FNAB material may provide new objective measures of the biologic potential of adrenal cortical carcinoma. The validity of these tests is enhanced with the use of image-based quantitative approaches that provide greater reproducibility and objectivity of their results.

Adrenal Cortex Neoplasms↗
PubMed1996

Relationship of genetic instability with immunoreactivities for p53 protein and proliferating cell nuclear antigen in transitional cell carcinoma of the bladder.

OBJECTIVE: To elucidate the relationship between genetic instability and tumor development in transitional cell carcinoma (TCC) of the bladder. METHODS: The genetic instability as assessed by 2c deviation index (2cDI) and 5c exceeding rate (5cER) was compared with immunoreactivities for p53 protein and proliferating cell nuclear antigen (PCNA) in specimens obtained from 65 patients with primary untreated TCCs. RESULTS: Both p53 and PCNA immunoreactivities significantly correlated with histological grade as well as stage. The immunoreactivity of p53 significantly correlated with that of PCNA, while a dissociation of the positive correlation between p53 immunoreactivity and PCNA expression was noted in the TCCs with high 2cDI value (> or = 2.0) and/or high 5cER (> or = 10%). In addition, PCNA expression became higher as the genetic instability increased, however, the p53 immunoreactivity was not parallel with the change of genetic instability. Moreover, some TCCs with high genetic instability (2cDI > or = 2.0 and 5cER > or = 10%) showed concomitant expression of high PCNA and low p53 protein, whose clinical outcome was poor in general. CONCLUSIONS: The inactivation of p53 may represent a rather early event in the development of TCC of the bladder. However, the proliferative activity in itself rather than the effect due to a specific alteration in p53 plays an important role in the development and progression in TCC of the bladder.

Carcinoma, Transitional Cell↗
PubMed1999

Comparative histopathology of radial artery versus internal thoracic artery and risk factors for development of intimal hyperplasia and atherosclerosis.

BACKGROUND: In this study, we examined the comparative histopathology, morphometry, and risk factors for the development of intimal hyperplasia and atherosclerosis in the radial artery (RA) and the internal thoracic artery (ITA). METHODS AND RESULTS: Paired specimens of RAs and ITAs, obtained from 150 patients who underwent CABG, were evaluated with histopathology; 110 pairs of arteries were suitable for morphometric analysis. The severity of disease was evaluated on the basis of percentage of luminal narrowing, intimal thickness index, and intima-to-media ratio. Risk factors were determined with stepwise linear regression. Intimal hyperplasia was seen in 141 RAs (94%) and 103 ITAs (69%) (P<0.001). Atherosclerosis was seen in 5% of RAs and 0.7% of ITAs (P=0.04). Medial calcification was found only in RAs (20 of 150, 13.3%) (P<0.001). Morphometric analysis showed that compared with ITAs, RAs had a significantly higher intimal area, medial area, percentage of luminal narrowing, intimal thickness index, and intima-to-media ratio (all P<0.001) Factors found to be significant (P<0.05) predictors of the 3 severity indices of intimal hyperplasia, including atherosclerosis, in RAs were peripheral vascular disease, smoking, age, and diabetes. Risk factors for intimal hyperplasia in ITAs were age and smoking. CONCLUSIONS: The RA is more likely to have atherosclerosis, intimal hyperplasia, and medial calcification than the ITA. Morphometric analysis indices showed marked differences between the RA and the ITA. Care should be taken when selecting the RA as a conduit in CABG, particularly in patients who are elderly, diabetic, smoke, or have peripheral vascular disease.

Adult↗
PubMed1996

Effects of lisinopril on the structure of renal arterioles.

We investigated the effect of long-term administration of the angiotensin-converting enzyme inhibitor lisinopril on renal arterioles in spontaneously hypertensive rats (SHR) and Wistar-Kyoto rats (WKY) using a morphometric method and vascular cast technique. Rats were treated with lisinopril beginning at 4 weeks of age. At 15 weeks of age, the kidney vessels were fixed when maximally relaxed. Resin was perfused into the right kidney to make a cast of the renal vasculature. The opposite kidney was used for the morphometric study to evaluate structural changes of the vascular wall. The vascular cast study demonstrated a significant reduction in the lumen diameter of the afferent but not the efferent arterioles in SHR compared with those in WKY. In lisinopril-treated rats, the afferent arteriolar lumen diameters were significantly larger than those of the respective control groups in both strains. However, treatment did not affect the lumen diameter of efferent arterioles in either strain. The morphometric study revealed that the cross-sectional area of afferent arteriolar media was significantly smaller in SHR than WKY, suggesting that the impaired growth of the afferent arteriolar media was involved in the narrowed afferent arteriolar lumen in SHR. The presence of significantly smaller media-lumen ratio, greater media cross-sectional area, and larger internal as well as external diameters of the afferent arterioles in treated SHR than in untreated rats suggested that lisinopril treatment normalizes the structure of the afferent arterioles in SHR by vascular reverse remodeling and by inducing media growth.

Angiotensin-Converting Enzyme Inhibitors↗
PubMed2002

Platelets activated by collagen through immunoreceptor tyrosine-based activation motif play pivotal role in initiation and generation of neointimal hyperplasia after vascular injury.

BACKGROUND: Platelet adhesion on components of the extracellular matrix and platelet activation by those components are crucial for the arrest of posttraumatic bleeding, but they can also harm tissue by occluding diseased vessels. Recent studies have shown that the activation of platelets by collagen is mediated through the same pathway used by immune receptors, with an immunoreceptor tyrosine-based activation motif on the Fc receptor gamma chain (FcRgamma) playing a pivotal role. METHODS AND RESULTS: We examined the role of collagen-stimulated platelets in the development of injury-induced neointimal formation by using mice deficient in FcRgamma. The left femoral arteries of 8- to 12-week-old FcRgamma-deficient mice (n=16) and C57BL/6 (wild-type) mice (n=16) were injured by a straight spring wire (0.35-mm diameter). Segments of the injured and uninjured femoral arteries were excised at 7 days and 28 days after the vascular injury. Arterial segments were examined by immunohistochemistry and electron microscopy. Two hours after injury, electron microscopy showed marked decreases in platelet adhesion and neutrophil attachment to the vascular wall surface in FcRgamma-knockout mice compared with wild-type mice. At 7 days after injury, staining with anti-neutrophil antibody showed fewer neutrophils in FcRgamma-knockout mice than in wild-type mice. Computer-aided morphometry performed to measure the neointimal area, intima/media ratio, and stenotic area at 28 days after injury showed a significantly smaller ratio and area in FcRgamma-knockout mice than in wild-type mice (for neointimal area, 16 635 +/- 1406 versus 31 483 +/- 2309 microm2, respectively; for intima/media ratio, 1.25 +/- 0.40 versus 2.68 +/- 0.04, respectively; and for stenotic area, 26.8 +/- 2.1% versus 49.3 +/- 4.1%, respectively). CONCLUSIONS: These results demonstrate that FcRgamma may play important roles in the initiation and generation of neointimal hyperplasia after balloon injury through the activation of platelets by collagen.

Adaptor Proteins, Signal Transducing↗
PubMed2005

Basal-like cells constitute the proliferating cell population in cystic fibrosis airways.

RATIONALE: Cystic fibrosis airways are recurrently exposed to noxious stimuli, leading to epithelial injury. Previous reports suggest that cystic fibrosis airway epithelia may respond to injury by increasing proliferation. OBJECTIVES: We sought to determine the characteristics of the proliferating cell population in cystic fibrosis airways. METHODS: Six cystic fibrosis and six normal lung sections from lung transplant recipients or lung surgery were obtained from the Duke Hospital pathology archives. Sections containing bronchi were evaluated for epithelial cell proliferation using immunohistochemistry for a nuclear proliferation antigen, Ki-67, and image analysis; immunohistochemistry for basal cells using a cytokeratin 5/14 antibody; and immunohistochemistry for the epidermal growth factor receptor and ErbB2, two receptor tyrosine kinases implicated in epithelial proliferation and differentiation. RESULTS: Overall, cystic fibrosis sections had a greater proliferation index than control sections with 25.1 +/- 2.1% positively staining nuclei/total nuclei compared with control sections, 4.6 +/- 0.9% (p = 0.002). In cystic fibrosis sections only, there were areas of hyperplastic cuboidal cells adjacent to normal pseudostratified columnar epithelial sections; in these areas of epithelial hyperplasia, there was uniform Ki-67 staining, indicating a zone of proliferating cells. The proliferating cell population also expressed the basal cell cytokeratins 5/14 and epidermal growth factor receptor. Expression of ErbB2 was diminished in the proliferating cells. CONCLUSIONS: Our results suggest that basal-like cells, expressing the epidermal growth factor receptor, constitute the proliferating cell population in cystic fibrosis airways.

Adolescent↗
PubMed1996

Expression of laminins in the airways in various types of asthmatic patients: a morphometric study.

Laminins (Ln) are crucial in airway morphogenesis. Because they are able to interact with inflammatory cells, they are likely to participate in inflammation accompanied by airway structural remodeling in asthma. Taking biopsies and using immunohistochemistry and quantitative image analysis, we characterized the distribution of Ln chains alpha 1, alpha 2, and beta 2 in the bronchial mucosa of patients with seasonal (n = 17), early occupational (n = 8), and chronic asthma (n = 16) for comparison with that of normal controls (n = 8). In all asthmatic patients, both Ln chains alpha 1 and beta 2 were confined to the superficial margin of the basement membrane (BM), blood vessels, and smooth muscle. The thickness of Ln beta 2 expression in BM was significantly greater in patients with chronic (1.9 +/- 0.1 microns; P < 0.001) and occupational asthma (1.7 +/- 0.1 microns; P < 0.05) than in controls (0.4 +/- 0.3 microns). Only in patients with occupational asthma was the thickness of the Ln alpha 1 layer (2.3 +/- 0.2 microns; mean +/- SEM) significantly different from that in controls (1.4 +/- 0.5 microns; P < 0.05). There was no immunoreactivity for the Ln alpha 2 chain in controls or patients with mild asthma, but in clinically severe chronic asthma we found a discontinuous staining along the epithelial margin of the BM. Since Ln chains alpha 2 and beta 2 appear to function only during morphogenesis, increased expression of these Ln chains in adult asthma patients suggests accelerated tissue turnover in the airways, possibly as a result of airway inflammation in asthma.

Adult↗
PubMed1997

Role of DNA measurements of head and neck tumors.

The measurement of DNA ploidy has been used as a tool to try to determine the prognosis of many neoplasms. DNA ploidy can be determined by flow cytometry or image analysis of a tumor. In squamous carcinomas of the head and neck, a poorer prognosis has been associated with nondiploid tumors. Similar results have been obtained from studies of salivary gland neoplasms. The role of DNA ploidy as a provider of independent information has yet to be determined. With rarer head and neck tumors, measurement of tumor ploidy has yet to be established as a valuable adjunct to routine light microscopic study.

Carcinoma, Squamous Cell↗
PubMed2001

Texture analysis of fluorescence lifetime images of AT- and GC-rich regions in nuclei.

We used intensity and fluorescence lifetime microscopy (FLIM) of 3T3 nuclei to investigate the existence of AT-rich and GC-rich regions of the nuclear DNA. Hoechst 33258 (Ho) and 7-aminoactinomycin D (7-AAD) were used as fluorescence probes specific for AT and GC base pairs, respectively. YOYO-1 (Yo) was used as a dye that displays distinct fluorescence lifetimes when bound to AT or GC base pairs. We combined fluorescence imaging of Ho and 7-AAD with time-resolved measurements of Yo and took advantage of an additional information content of the time-resolved fluorescence. Because a single nucleus could not be stained and measured with all three dyes, we used texture analysis to compare the spatial distribution of AT-rich and GC-rich DNA in 100 nuclei in different phases of the cell cycle. The fluorescence intensity-based analysis of Ho- or 7-AAD-stained images indicates increased number and larger size of the DNA condensation centers in the G2/M-phases compared to G0/1-phases. The lifetime-based study of Yo-stained images suggests spatial separation of the AT- or GC-rich DNA regions in the G2/M-phase. Texture analysis of fluorescence intensity and lifetime images was used to quantitatively study the spatial change of condensation and separation of AT- and GC-rich DNA during the cell cycle.

3T3 Cells↗