Miljana Pavlović

Morphometric Analysis of Nonsclerosed Glomeruli Size and Connective Tissue Content during the Aging Process

Number of sclerotic glomeruli increases during the aging process. Consequently, majority of remained nonsclerosed glomeruli become hypertrophic and some of them sclerotic, too. The aim of this study was to quantify the size and connective tissue content of nonsclerosed glomeruli and to evaluate the percentage of hypertrophic ones in examined human cases during the aging. Material was right kidneys tissue of 30 cadavers obtained during routine autopsies. Cadavers were without previously diagnosed kidney disease, diabetes, hypertension, or any other systemic disease. Tissue specimens were routinely prepared for histological and morphometric analysis. Images of the histological slices were analyzed and captured under 400x magnification with digital camera. Further they were morphometrically and statistically analyzed with ImageJ and NCSS-PASS software. Multiple and linear regression of obtained morphometric parameters showed significant increase of glomerular connective tissue area and percentage. Cluster analysis showed the presence of two types of glomeruli. Second type was characterized with significantly larger size, connective tissue content, and significantly lower cellularity, in relation to the first type. Such glomeruli might be considered as hypertrophic. First type of glomeruli was predominant in younger cases, while second type of glomeruli was predominant in cases older than 55 years.

Analysis of Fascicular Structure and Connective Tissue Sheaths in Sural Nerve during Aging

Summary The aim of our study was to analyze the changes of connective tissue sheaths of epi-, peri- and endoneurium of sural nerve during aging. The study was conducted on sural nerve samples of 10 cases aged 9-80 years. The specimens were embedded in paraffin using standard procedures, after which 5-μm-thick cross-sections of nerve trunks were made and stained using Masson’s trichrome staining. After morphological analysis of fascicular structure and connective sheaths of the nerve, morphometric analysis was conducted using the software for digital image analysis “ImageJ”. Each investigated case was analyzed for total neural, epineurial and fascicular cross-section area, mean values of perineurial index, volume density of myelinated axons and of endoneurial content. To test the difference in mean values for statistical significance we used the Student’s T-test for small independent sample. The number of fascicles was 5-13, while the majority of the nerves had less than 10 fascicles. Fascicular structure, which included the number of fascicles and epifascicular/fascicular area ratio, did not show significant changes during aging. Perineurial thickness /fascicle size ratio statistically significantly increased in the older investigated group (p<0.05). Myelinated fibres were of smaller diameter, with more irregular form and markedly less frequent in older cases. Quantitative analysis showed statistically significant decrease in volume density of myelinated fibres in the older group. As results of applied investigation methods we found thickening of perineurial sheath of sural nerve during aging, as well as endoneurial fibrosis. Future investigations of age-related changes should focus on analysis of the components of extracellular matrix within perineurium and endoneurium.

Longitudinal striae of the human fornix: shape, relations and variations

BackgroundTwo pairs of sagittal longitudinal striae, medial and lateral, are slender bundles of fibers located on the dorsal surface of corpus callosum, situated deeply in the longitudinal fissure of telencephalon. Imbedded in the structure of tiny gyrus, indusium griseum, they are, in fact, supracallosal fibers of the fornix, previously called fornix longus.MethodsLongitudinal striae were investigated in 25 fixed human brains obtained from autopsies. Macrodissection and morphometric methods were used in order to find out and analyze the appearance and gross morphological variability of longitudinal striae, as well as their inter-individual relations.ResultsLateral longitudinal striae were located along the sulci of corpus callosum. Medial striae were positioned along the sagittal midline and they were mostly individual. However, they were at times connected, spanned or duplicated. Longitudinal striae make a characteristic pattern on the dorsal surface of corpus callosum. A classification of striae is made on the basis of their appearance.ConclusionsAlthough similar at first sight the striae, especially medial ones, have some individual features which make the pattern variable. Medial striae are more variable than the lateral ones. Perhaps functional neuroimaging and DT MRI will disclose the enigma of these striae.

Morphometric analysis of the human anterior pituitary's folliculostellate cells during the aging process

Folliculostellate cells represent non-endocrine cells of the anterior pituitary which influence the function of the endocrine cells via paracrine action. Though there is a lack of literature data on their presence during human aging, the aim of this research was to perform the quantification of anterior pituitary folliculostellate cells by the application of immunohistochemical and morphometric methods. The material for the study consisted of 15 anterior pituitaries taken from cadavers at routine autopsy. Their tissue was processed by standard histological procedure and the obtained histological slices were stained by S100 polyclonal antibody. Digital images of stained histological sections were analyzed by morphometric method with ImageJ system. The volume density of S100 positive cells was measured for each case. Results of morphometric and statistical analysis showed a significantly positive correlation between folliculostellate cell volume density and the age of the evaluated cases. Linear regression additionally showed that the age significantly predicts folliculostellate cells volume density in our sample. Further, all cases were classified into three age groups and One Way ANOVA showed that the volume density of folliculostellate cells was significantly higher only in the third age group in relation to the first and the second group, respectively. Volume densities of the first and the second age groups were not significantly different. So, the results of our study pointed to the conclusion that folliculostellate cells presence generally increases with age, but this increase is significant only in the oldest cases and might represent the modality of successful anterior pituitary aging.

Parahippocampal corpora amylacea and neuronal lipofuscin in human aging

The aim of this research was to quantify the number of corpora amylacea and lipofuscin-bearing neurons in the parahippocampal region of the brain. Right parahippocampal gyrus specimens of 30 cadavers were used as material for histological and morphometric analyses. A combined Alcian Blue and Periodic Acid-Schiff technique was used for identification and quantification of corpora amylacea and lipofuscin-bearing neurons. Immunohistochemistry was performed using S100 polyclonal, neuron-specific enolase and glial fibrillary acidic protein monoclonal antibodies for differentiation of corpora amylacea and other spherical inclusions of the aging brain. Cluster analysis of obtained data showed the presence of three age groups (median age: I = 41.5, II = 68, III = 71.5). The second group was characterized by a significantly higher numerical density of subcortical corpora amylacea and number of lipofuscin-bearing neurons than other two groups. Values of the latter cited parameters in the third group were insignificantly higher than the first younger group. Linear regression showed that number of parahippocampal lipofuscin-bearing neurons significantly predicts numerical density of subcortical corpora amylacea. The above results suggest that more numerous parahippocampal region corpora amylacea and lipofuscin-bearing neurons in some older cases might represent signs of its’ neurons quantitatively-altered metabolism.

Morphometric Study of Connective Tissue in the Human Pituitary during Aging Process

SUMMARY The aim of the research was to quantify the connective tissue and blood vessels in the aging human pituitary, using the morphometric methods. The material contained 29 pituitary tissues from cadavers of both sexes (14 females and 15 males), aged 33-95 years. The pituitary tissue was processed by standard histological procedure and stained by Mallory trichrome method. Histological samples were analyzed using a light microscope with magnification of 400x. Morphometric analysis was performed on digital pictures with the resolution of 1.3 megapixels, at 40 pituitary visual fields of each case examined. Fovea Pro 4 system was used for the analysis. Above the digital images of each field of view, there was inserted a simple network test system. Using this programme, there were determined bulk densities of connective tissue and blood vessels in the examined cases. The results were statistically analyzed using the package NCSS-PASS 2007. Correlation analysis showed that the bulk density of connective tissue statistically significantly increased during aging. The average bulk density of blood vessels in the pituitary manifested statistically significant decrease during the aging process. These results were confirmed by the linear regression analysis. The average bulk density of the connective tissue, as well as the average bulk density of blood vessels in the pituitary cases, did not differ significantly among males and females. The process of aging is followed by the proliferation of the connective tissue and fibrosis of the pituitary, which is accompanied by the reduction of its vascular network. Sažetak Cilj istraživanja bio je da se morfometrijskom metodom kvantifikuje prisustvo vezivnog tkiva i prokrvljenost adenohipofize tokom starenja čoveka. Materijal je tkivo adenohipofize 29 kadavera oba pola (14 ženskih i 15 muških), starosti 33-95 godina. Tkivo hipofize obrađeno je standardnom histološkom procedurom i bojeno Mallory trihromnom metodom. Dobijeni histološki preparati analizirani su pomoću svetlosnog mikroskopa pod uvećanjem 400x. Morfometrijska analiza je obavljena na digitalnim fotografijama rezolucije 1.3 mega piksela, po 40 vidnih polja adenohipofize svakog od ispitanih slučajeva, uz pomoć Fovea Pro 4 sistema za analizu. Preko digitalne slike svakog vidnog polja superponiran je prosti mrežni testni sistem. Pomoću njega su određivane zapreminska gustina vezivnog tkiva (Vvt) i krvnih sudova (Vvks) ispitanih slučajeva, koje su izračunavane pomoću formule Kališnika (1985). Dobijeni rezultati su statistički analizirani korišćenjem paketa NCSS-PASS 2007. Korelaciona analiza je ukazala da zapreminska gustina vezivnog tkiva analiziranih slučajeva statistički značajno raste tokom procesa starenja. Prosečna zapreminska gustina krvnih sudova adenohipofize ispoljavala je statistički značajan pad tokom procesa starenja. Ovi rezultati su potvrđeni linearnom regresionom analizom. Prosečna zapreminska gustina vezivnog tkiva, kao i prosečna zapreminska gustina krvnih sudova adenohipofiza kod muškog i ženskog pola ne razlikuju se statistički značajno. Proces starenja praćen je proliferacijom vezivnog tkiva i fibrozom adenohipofize koja je praćena redukcijom njene vaskularne mreže.