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PubMed1992

A computer-assisted automatic method for myelinated nerve fiber morphometry.

This paper describes a computer-assisted automatic method developed for myelinated nerve fiber morphometry. IBAS image analysis system was used. One-micron thick sections of sural nerve were stained with p-phenylenediamine. Dark myelin sheaths, standing out against a pale background, were identified by a binary transformation of the computer-stored image. Different parameters of the fibers as well as of the axons can be measured. The automatic system allows operator-interactive manipulation, if required, to avoid inclusion of wrong histologic structures. A large spectrum of statistics facilities are available. We performed a comparison between automatic and semi-automatic analysis. Measurements resulted virtually overlapping, but time was three to four times shorter with automatic procedure. The method here described is accurate and reliable, and permits economy of time and effort.

Biopsy↗

PubMed1995

Nucleolar organizer regions and image analysis nuclear morphometry of small cell (nevoid) melanoma.

Small cell (nevoid) melanomas may provide difficulties in diagnosis as their constituent cell type resembles a benign nevoid melanocyte. In the present study, 10 small cell melanomas were analyzed for the silver staining of their nucleolar organizing regions (AgNORs), and their nuclear area and perimeter were measured by computerized digital image analysis and compared with 10 superficial spreading melanomas lacking small cell differentiation and 10 dermal nevi. The average number of AgNORs per nucleus was 5.83 (SD +/- 1.69) for small cell melanomas and was significantly different when compared with 8.49 (SD +/- 1.58) for superficial spreading melanomas (p < 0.05) and 2.71 (SD +/- 0.50) for dermal nevi (p < 0.05). Digital image analysis confirmed that the nuclear perimeter and nuclear area of cells in nevoid melanomas did not significantly differ from those of ordinary dermal nevi (p > 0.05), but both group were significantly different from superficial spreading melanomas lacking a small cell morphology (p < 0.05). Counting AgNOR numbers may be useful in evaluating small cell (nevoid) melanomas and provides a technique for differentiating their constituent cell from ordinary nevus cells. Nuclear morphometry determined by digital image analysis may help better define the nuclear size in small cell melanomas.

Adolescent↗

PubMed1996

Nuclear morphometry, immunohistochemical staining with Ki-67 antibody and mitotic index in the assessment of proliferative activity and prognosis of primary malignant melanomas of the skin.

Nuclear morphometry, immunohistochemical staining with Ki-67 antibody and mitotic index were studied in primary cutaneous malignant melanomas. The number of Ki-67 positive cells/ 200 tumor cells did not correlate with any nuclear morphometrical parameters, and it only approached but did not reach significant correlation with melanoma thickness according to Breslow. The nuclear area, short axis and long axis correlated with melanoma thickness, but the nuclear axis ratio (which reflects the sphericity of nuclei) and melanoma thickness did not show significant correlation. Mitotic index was higher in thick melanomas and in melanomas with high Ki-67 positivity, large nuclear area, long nuclear short axis, and small nuclear axis ratio. In Cox's stepwise proportional hazard model, melanoma thickness and the nuclear axis ratio were significant independent prognostic factors for patient survival, while the nuclear area, short axis and long axis, gender, age, Clark level, mitotic index and Ki-67 positivity lacked significant independent prognostic value. The results suggest that the proliferative activity of tumor cells does not alone explain the great importance of tumor thickness as prognosticator in melanoma. The thickness of melanoma measured according to Breslow and the nuclear axis ratio are more efficient prognosticators in melanoma than parameters associated with proliferation.

Adolescent↗

PubMed1989

Qualitative assessment and morphometry in the study of the ileal reservoir after restorative proctocolectomy.

Little is known about the long-term effects on the reservoir mucosa in patients with ulcerative colitis and familial polyposis coli who undergo proctocolectomy with subsequent construction of ileal reservoir/pouch and ileoanal anastomosis. In these patients, questions regarding adaptation towards a more colon-like mucosa and/or development of (pre)malignant changes are of particular importance. With the aim of designing a method for reliable evaluation of the mucosa in the ileal pouch, biopsies from 10 patients were studied by semiquantitative assessment and morphometry. The findings were compared with those obtained from normal jejunum, ileum, and colon. The following parameters were found to be important: Villous surface density, quantity of goblet cells, number of mitoses, and the presence/absence of predominantly sulphated mucin+ goblet cells. The number of Paneth cells did not show significant changes. The villous surface density was determined by a cycloid test system applied to vertical sections. Semiquantitative assessment was a sufficiently precise method for the evaluation of the quantity of goblet cells. The counting of sulphated mucin+ goblet cells was not reproducible, instead a simple statement about the presence or absence of these cells was judged to be adequate. The number of mitoses and of Paneth cells were counted directly. During the first year of function the ileal pouch showed signs of adaptation towards a colon-like mucosa: Reduction of villous surface density, increased mitotic activity, and appearance of sulphated mucin+ goblet cells. The number of Paneth cells did not show significant changes. The amount of goblet cells was generally not increased, rather reduced in some patients.

Adult↗

PubMed1991

Gross examination of musculoskeletal sarcomas. Estimation of tumour volume and sampling for unbiased morphometry.

A method for handling and gross examination of specimens from musculoskeletal sarcomas is described. The method, which can be modified to relate to modern, preoperative tumour imaging, allows an accurate estimation of tumour volume and a systematic random sampling for morphometry, which for instance permits the unbiased estimation of tumour necrosis, cellularity and mitotic index. Furthermore, sampling can be performed in such a way that stereological parameters (for the estimation of nuclear size and pleomorphism) based on 'vertical sections' can be measured. The method in no way prohibits routine sampling, for instance from specimen margins and specific areas of interest within the tumour, and with a little practice does not take much extra time.

Bone Neoplasms↗

PubMed1996

Differential effects of prenatal exposure to cocaine and amphetamine on growth parameters and morphometry of the prefrontal cortex in the rat.

The purpose of this study was to investigate the differential effects of prenatal exposure to psychostimulants, e.g., cocaine or amphetamine, on basic growth parameters and morphometry of the medial prefrontal cortex of the rat. A group of pregnant Wistar rats was given 60 mg/kg body weight/day of cocaine hydrochloride and another group 10 mg/kg body weight/day of d-amphetamine sulfate, subcutaneously, from gestational days 8 to 22. Control groups of pregnant rats were pair-fed; litters were culled to eight pups (4 males and 4 females) weighed every other day until postnatal day 30 and every week until day 90. The body weight growth patterns modelled by a Gompertz curve were different in rats prenatally exposed to the two psychostimulants. Rats exposed to amphetamine had on average a slower growth than those exposed to cocaine, reaching an identical estimated adult weight. Allometric relationships between forebrain and body weight and cerebellum and body weight were described by two distinct postnatal growth phases that are different among the experimental groups. In the comparison of the two psychostimulants the relative cerebellum/body growth is lower in the offspring of the cocaine group than in the amphetamine group between PND14-PND30; between PND30-PND90 the relative growth rate is considerably higher in the offspring of the cocaine dams compared to that of the amphetamine dams. Groups of perfused animals were selected at postnatal days 14 and 30 to analyze the morphometric organization of the medial prefrontal cortex. In serial celloidin sections the volumes of the prefrontal cortex were determined; the number of neurons per unit volume of reference area was calculated using the stereological technique of the disector. The changes found in the morphometric parameters show a catch-up at postnatal day 30 of the "increased" density of neurons of the medial prefrontal cortex found at postnatal day 14. These data show differential growth patterns of offspring from cocaine- and amphetamine-exposed rats; a delayed development in the achievement of normal morphometric parameters of neurons in the prelimbic subarea of the medial prefrontal cortex occurs in the prenatally amphetamine-exposed offspring at early ages, and a catch-up is found after the first month of life. Complementary studies are needed to assess whether these changes have functional implications in the rats exposed prenatally to psychostimulants.

Amphetamine↗

PubMed1994

Early morphometry of a retinal dystrophy in Labrador retrievers.

Retinal morphometry was assessed in 7 dogs from a colony of Labrador Retrievers with dystrophic retinas at 1,2,3,4 and 18 months of age. Rod outer segment length and outer nuclear layer width were measured in the central, midperipheral and peripheral retina at six locations along the vertical meridian. Early striking regional differences in onset and rate of progression were characteristic for this inherited retinal degeneration. Notably, some areas of the retina developed fully and normally before degenerating. The central parts of the vertical meridians showed slightly disorganized rod outer segments already at 1 month of age and they were significantly shorter than those of control animals at 3 and 4 months (p < 0.01 and p < 0.001, respectively). The rod outer segments of the midperipheral and peripheral regions were, however, comparable to control animals as late as at 4 months of age. At 18 months the rod outer segments of dystrophic animals were significantly shorter in all retinal regions (p < 0.0005). At the age the outer nuclear layer of the dystrophic animals had become significantly thinner than that of control animals in all retinal regions (p < 0.001), indicating a clear visual cell loss. It is reasonable to characterize this as a retinal degeneration having a relatively slow progression, which enhances its relevance to conditions of clinical significance.

Aging↗

PubMed2002

Alveolar fibrosis and changes in equine lung morphometry in response to intrapulmonary blood.

Necropsy studies of horses suffering exercise-induced pulmonary haemorrhage (EIPH) have identified mild inflammatory lesions with evidence of alveolar fibrosis and bronchiolitis. These lesions were thought to be the result of viral infections that predisposed the affected regions of the lung to EIPH. We have shown previously that during erythrophagocytosis in the alveolar space, there is a prolonged period of macrophage influx and activation. This present study used morphometric analysis to quantify the effects of macrophage activity during erythrophagocytosis, on the alveolar cell population and physical structure of the alveolar walls. Segments of the bronchial tree were inoculated with either autologous whole blood or serum, at 15, 8, 3 days, 24 h and 30 min prior to euthanasia. Blood inoculation produced many significant changes in the alveolar morphometry including, increased numbers of alveolar macrophages, increased septal thickness, and a markedly increased percentage of collagen in the alveolar walls. Signs of chronic inflammation including increased macrophage activity and erythrophagocytosis coincided with increased alveolar macrophage numbers (10,688 +/- 1708 cells/mm3 to 30,957 +/- 6831 cells/mm3), septal thickness (4.1 +/- 0.4 microm to 6.1 +/- 0.5 microm) and alveolar septal collagen content (6.6 +/- 0.5% to 27.5 +/- 3.3%). The results suggest that intrapulmonary blood induces a macrophage dominated inflammatory response, septal thickening and the development of alveolar fibrosis. These changes are the probable cause of the observed alveolar fibrosis and bronchiolitis that was once suspected to be the originating cause of EIPH.

Animals↗

PubMed1988

Automated morphometry of corneal endothelial cell: use of video camera and video tape recorder.

We developed an apparatus for automated morphometry of the corneal endothelium, which was photographed through a specular microscope connected to a video camera, and the images were stored on a video tape. The clearest stationary image was input into an image analyser to determine automatically the cell boundaries. Although human interaction is generally necessary, the mean time required to complete this procedure was about 13 minutes, based on the results of the 30 normal eyes, and the time needed for manual correction was about 4 minutes. The mean cell area obtained by this method correlated well (r = 0.9335) with that obtained by tracing the same images. This apparatus is clinically useful for immediately obtaining the mean cell area of corneal endothelium and will extend the application of specular microscopy to the routine clinical setting.

Adult↗

PubMed1998

Dietary fibre and intestinal microflora: effects on intestinal morphometry and crypt branching.

BACKGROUND: Fermentable dietary fibre has many effects on the gastrointestinal tract. One is to alter epithelial crypt cell proliferation, especially in the colon. A discrepancy between epithelial cell production rates and intestinal weights has been noted previously: crypt cell production rates only increase if bacterial fermentation occurs, but intestinal wet weight can increase in the same animals without bacterial fermentation of fibre. AIMS: To quantify intestinal cell populations in order to resolve the above paradox. METHODS: Conventional and germ-free rats were fed fibre-free or fibre supplemented diets and their intestines were quantified by morphometry. RESULTS: There was evidence of fibre associated muscle hypertrophy in the colon, but the main effect of fibre was an increase in the number of crypts per circumference and also the number of branched crypts in the proximal colon in both groups. There was also a large increase in the number of branched crypts in the mid colon of the germ-free rats (both fibre-free and fibre supplemented). Fibre had a direct (bacteria independent) effect on goblet cells in the small intestine and a direct effect on the goblet cells in the colon, which was attenuated by the presence of bacteria. There was a notable decline in the number of enteroendocrine cells in the small intestine of the germ-free animals. CONCLUSIONS: Fibre has several direct and indirect effects on the gut. In the proximal colon it can directly increase the number of crypts present. This provides a means for increasing intestinal mass in addition to intestinal crypt cell production.

Animals↗

PubMed1998

HIV enteropathy: comparative morphometry of the jejunal mucosa of HIV infected patients resident in the United Kingdom and Uganda.

AIMS: To compare jejunal mucosal morphometry in HIV infected patients resident in London and Uganda. PATIENTS: Twenty HIV positive patients from London and 16 from Uganda were studied, and compared with HIV negative control subjects from both sites. METHODS: Stools and biopsy specimens were examined for enteropathogens. Surface area to volume (S:V) ratio was estimated morphometrically, mean crypt length of jejunal biopsy specimens was measured, and HIV infected cells detected immunohistochemically were quantified. RESULTS: Enteric pathogens were detected in none of the London patients, and in three Ugandan patients. S:V ratio was lower, and mean crypt length higher, in the specimens of London patients than in normal subjects, but there was no difference in S:V ratio or mean crypt length between Ugandan patients and controls. A negative correlation was present between S:V ratio and mean crypt length in all biopsy specimens analysed. HIV infected cells were detected only in lamina propria. CONCLUSION: Infection of cells in the lamina propria of the jejunum with HIV stimulates crypt cell proliferation, and a fall in villous surface area. The mucosal response to HIV is masked by other pathogens in the African environment.

Adult↗

PubMed2006

Nuclear morphometry in columnar cell lesions of the breast: is it useful?

AIMS: To evaluate the nuclear morphometric features of breast columnar cell lesions (CCLs) observed on mammotome core biopsies, to determine if there are significant measurable differences between those with atypia and those without. Correlation with follow-up open excision specimens was made. METHODS: Mammotome core biopsies performed on patients that contained CCLs were derived from the departmental case files. Histological material was reviewed and foci of CCLs demarcated for nuclear morphometric assessment, which was accomplished using an imaging system. Nuclear parameters studied were nuclear area and perimeter, circularity factor and feret's diameter. Statistical analysis used the GraphPad Prism software, with p<0.05 indicating significance. RESULTS: On examination of core biopsies of 40 patients with CCLs, 8 lesions were benign, 4 showed atypical lobular hyperplasia, 8 showed CCLs with nuclear atypia, 19 disclosed atypical ductal hyperplasia (ADH) and 1 showed ductal carcinoma in situ (DCIS). The nuclear area, perimeter and feret's diameter of CCLs with atypia were significantly greater than those without (p = 0.04, 0.03 and 0.019, respectively), whereas no difference was observed in the circularity factor. Follow-up open excision biopsy specimens in 24 patients showed upgrading to DCIS in 40% of cases diagnosed initially with ADH on core biopsy compared with 20% of CCLs with atypia. CONCLUSIONS: Nuclear morphometry in CCLs confirms nuclear size as the key parameter in the assessment of nuclear atypia. Whether it can be potentially used as an adjunctive tool depends on the establishment of appropriate cut-offs.

Adult↗

PubMed1994

Facial morphometry of Ecuadorian patients with growth hormone receptor deficiency/Laron syndrome.

Facial morphometry using computerised image analysis was performed on patients with growth hormone receptor deficiency (Laron syndrome) from an inbred population of southern Ecuador. Morphometrics were compared for 49 patients, 70 unaffected relatives, and 14 unrelated persons. Patients with growth hormone receptor deficiency showed significant decreases in measures of vertical facial growth as compared to unaffected relatives and unrelated persons with short stature from other causes. This report validates and quantifies the clinical impression of foreshortened facies in growth hormone receptor deficiency.

Adolescent↗

PubMed2005

A voxel based morphometry study on mild cognitive impairment.

BACKGROUND: Mild cognitive impairment (MCI) is the most widely used concept in classifying cognitive impairment in the elderly who do not fulfil the criteria for dementia. MCI is considered to confer an increased risk of progressing to dementia and most often Alzheimer's disease (AD). Various approaches such as imaging of the brain have been applied to predict the conversion of MCI to dementia. A number of volumetric magnetic resonance imaging (MRI) studies have detected atrophy of the medial temporal lobe in subjects with MCI, but for the other cerebral regions the results have been inconsistent. OBJECTIVE: To study the pattern of brain atrophy in MCI. METHODS: Thirty two controls and 51 individuals with MCI deriving from population based cohorts were studied by MRI using voxel based morphometry. The threshold of t maps was set at p < 0.001. RESULTS: Individuals with MCI had significant unilateral atrophy in the medial temporal lobe on the right side. Less extensive atrophy was found elsewhere-for example, in the temporal lobe, left superior parietal lobule, left anterior cingulate gyrus, and bilaterally in the thalami. CONCLUSIONS: The MRI findings in MCI resemble those seen in early AD.

Aged↗

PubMed2005

Traumatic brain injury and grey matter concentration: a preliminary voxel based morphometry study.

BACKGROUND: Magnetic resonance imaging (MRI) studies have shown diffuse cerebral atrophy following traumatic brain injury. In the past, quantitative volumetric analysis of these changes was carried out by manually tracing specific regions of interest. In contrast, voxel based morphometry (VBM) is a fully automated technique that allows examination of the whole brain on a voxel by voxel basis. OBJECTIVE: To use VBM to evaluate changes in grey matter concentration following traumatic brain injury. METHODS: Nine patients with a history of traumatic brain injury (ranging from mild to severe) about one year previously were compared with nine age and sex matched healthy volunteers. T1 weighted three dimensional MRI images were acquired and then analysed with statistical parametric mapping software (SPM2). The patients with traumatic brain injury also completed cognitive testing to determine whether regional grey matter concentration correlated with a measure of attention and initial injury severity. RESULTS: Compared with controls, the brain injured patients had decreased grey matter concentration in multiple brain regions including frontal and temporal cortices, cingulate gyrus, subcortical grey matter, and the cerebellum. Decreased grey matter concentration correlated with lower scores on tests of attention and lower Glasgow coma scale scores. CONCLUSIONS: Using VBM, regions of decreased grey matter concentration were observed in subjects with traumatic brain injury compared with well matched controls. In the brain injured patients, there was a relation between grey matter concentration and attentional ability.

Adult↗

PubMed2005

Progression of structural neuropathology in preclinical Huntington's disease: a tensor based morphometry study.

BACKGROUND AND OBJECTIVES: Regional cerebral atrophy occurs in carriers of the Huntington's disease (HD) gene mutation before clinical diagnosis is possible. The current inability to reliably measure progression of pathology in this preclinical phase impedes development of therapies to delay clinical onset. We hypothesised that longitudinal statistical imaging would detect progression of structural pathology in preclinical carriers of the HD gene mutation, in the absence of measurable clinical change. METHODS: Thirty subjects (17 preclinical mutation positive, 13 mutation negative) underwent serial clinical and magnetic resonance imaging (MRI) assessments over an interval of 2 years. Statistically significant changes in regional grey and white matter volume on MRI were analysed using tensor based morphometry (TBM). This technique derives a voxel-wise estimation of regional tissue volume change from the deformation field required to warp a subject's early to late T1 images. RESULTS: Over 2 years, there was progressive regional grey matter atrophy in mutation-positive relative to negative subjects, without significant clinical progression of disease. Significant grey matter volume loss was limited to bilateral putamen and globus pallidus externa (GPe), left caudate nucleus, and left ventral midbrain in the region of the substantia nigra. CONCLUSIONS: While these results are consistent with previous cross sectional pathologic and morphometric studies, significant progression of atrophy in HD before the onset of significant clinical decline is now demonstrable with longitudinal statistical imaging. Such measures could be used to assess the efficacy of potential disease modifying drugs in slowing the progression of pathology before confirmed clinical onset of HD.

Adult↗

PubMed2005

Diffusion tensor imaging and voxel based morphometry study in early progressive supranuclear palsy.

BACKGROUND: A comprehensive characterisation of grey and white matter changes in progressive supranuclear palsy (PSP), the second most common extrapyramidal syndrome after Parkinson disease, is still not available. OBJECTIVE: To evaluate grey and white matter changes in mild PSP patients by voxel based morphometry (VBM) and diffusion tensor imaging (DTI), respectively. METHODS: 14 mild PSP patients and 14 healthy controls entered the study and underwent a clinical and neuropsychological evaluation according with a standardised assessment. Each subject had a structural magnetic resonance imaging (MRI) study. Processing analysis of MRI data was carried out according to optimised VBM and fractional anisotropy was determined. RESULTS: Compared with the controls, in PSP patients VBM analysis showed a significant clusters of reduced grey matter in premotor cortex, frontal operculum, anterior insula, hippocampus, and parahippocampal gyrus, bilaterally. With regard to subcortical brain regions, the pulvinar, dorsomedial and anterior nuclei of the thalamus, and superior and inferior culliculum were affected bilaterally. A bilateral decrease in fractional anisotropy in superior longitudinal fasciculus, anterior part of corpus callosum, arcuate fascicolus, posterior thalamic radiations, and internal capsule, probably involving the cortico-bulbar tracts, was present in PSP patients. CONCLUSIONS: These data provide evidence for both grey and white matter degeneration in PSP from the early disease stage. These structural changes suggest that atrophy of cortical and subcortical structures and neurodegeneration of specific fibre tracts contribute to neurological deficits in PSP.

Aged↗

PubMed2006

A magnetic resonance imaging study of patients with Parkinson's disease with mild cognitive impairment and dementia using voxel-based morphometry.

BACKGROUND: Dementia is common in Parkinson's disease, but the underlying brain pathology is not yet fully understood. AIM: To examine the changes in the brain of patients with Parkinson's disease with mild cognitive impairment (MCI) and dementia, using structural magnetic resonance imaging. METHODS: Using voxel-based morphometry, the grey matter atrophy on brain images of patients with Parkinson's disease and dementia (PDD; n = 16) and Parkinson's disease without dementia (PDND; n = 20), and healthy elderly subjects (n = 20) was studied. In the PDND group, 12 subjects had normal cognitive status and 8 had MCI. Standardised rating scales for motor, cognitive and psychiatric symptoms were used. RESULTS: Widespread areas of cortical atrophy were found in patients with PDD compared with normal controls (in both temporal and frontal lobes and in the left parietal lobe). Grey matter reductions were found in frontal, parietal, limbic and temporal lobes in patients with PDD compared with those with PDND. In patients with PDND with MCI, areas of reduced grey matter in the left frontal and both temporal lobes were found. CONCLUSION: These findings show that dementia in Parkinson's disease is associated with structural neocortical changes in the brain, and that cognitive impairment in patients with PDND may be associated with structural changes in the brain. Further studies with larger groups of patients are needed to confirm these findings.

Aged↗