Morphometric analysis of somatotropic cells of the adenohypophysis and muscle fibers of the psoas muscle in the process of aging in humans.

The aim of this research was to quantify changes of the adenohypophyseal somatotropes and types 1 and 2 muscle fibers with aging, as well as to establish mutual interactions and correlations with age. Material was samples of hypophysis and psoas major muscle of 27 cadavers of both genders, aged from 30 to 90 years. Adenohypophyseal and psoas major tissue sections were immunohistochemically processed and stained by anti-human growth hormone and anti-fast myosin antibodies, respectively. Morphometric analysis was performed by ImageJ. Results of morphometric analysis showed a significant increase in the somatotrope area, and significant decrease in somatotrope volume density and nucleocytoplasmic ratio with age. Cross-sectional areas of types 1 and 2, and volume density of type 2 muscle fibers decreased significantly with age. One Way ANOVA showed that the latter cited changes in the somatotropes and types 1 and 2 muscle fibers mostly become significant after the age of 70. Significant positive correlation was observed between the area of the somatotropes and volume density of type 2 muscle fibers. A significant negative correlation was detected between the nucleocytoplasmic ratio of the somatotropes and cross-sectional areas of types 1 and 2 muscle fibers. So, it can be concluded that after the age of 70, there is significant loss of the anterior pituitary's somatotropes associated with hypertrophy and possible functional decline of the remained cells. Age-related changes in the somatotropes are correlated with the simultaneous atrophy of type 1, as well as with the atrophy and loss of type 2 muscle fibers.

Morphometric analysis of connective tissue sheaths of sural nerve in diabetic and nondiabetic patients.

One of the most common complications of diabetes mellitus is diabetic neuropathy. It may be provoked by metabolic and/or vascular factors, and depending on duration of disease, various layers of nerve may be affected. Our aim was to investigate influence of diabetes on the epineurial, perineurial, and endoneurial connective tissue sheaths. The study included 15 samples of sural nerve divided into three groups: diabetic group, peripheral vascular disease group, and control group. After morphological analysis, morphometric parameters were determined for each case using ImageJ software. Compared to the control group, the diabetic cases had significantly higher perineurial index (P < 0.05) and endoneurial connective tissue percentage (P < 0.01). The diabetic group showed significantly higher epineurial area (P < 0.01), as well as percentage of endoneurial connective tissue (P < 0.01), in relation to the peripheral vascular disease group. It is obvious that hyperglycemia and ischemia present in diabetes lead to substantial changes in connective tissue sheaths of nerve, particularly in peri- and endoneurium. Perineurial thickening and significant endoneurial fibrosis may impair the balance of endoneurial homeostasis and regenerative ability of the nerve fibers. Future investigations should focus on studying the components of extracellular matrix of connective tissue sheaths in diabetic nerves.

Asymmetries in numerical density of pyramidal neurons in the fifth layer of the human posterior parietal cortex.

BACKGROUND/AIM: Both superior parietal lobule (SPL) of dorsolateral hemispheric surface and precuneus (PEC) of medial surface are the parts of posterior parietal cortex. The aim of this study was to determine the numerical density (Nv) of pyramidal neurons in the layer V of SPL and PEC and their potential differences. METHODS: From 20 (40 hemispheres) formaline fixed human brains (both sexes; 27- 65 years) tissue blocks from SPL and PEC from the left and right hemisphere were used. According to their size the brains were divided into two groups, the group I with the larger left (15 brains) and the group II with the larger right hemisphere (5 brains). Serial Nissl sections (5 microm) of the left and right SPL and PEC were used for stereological estimation of Nv of the layer V pyramidal neurons. RESULTS: Nv of pyramidal neurons in the layer V in the left SPL of brains with larger left hemispheres was significantly higher than in the left SPL of brains with larger right hemisphere. Comparing sides in brains with larger left hemisphere, the left SPL had higher Nv than the right one, and then the left PEC, and the right SPL had significantly higher Nv than the right PEC. Comparing sides in brains with the larger right hemisphere, the left SPL had significantly higher Nv than left PEC, but the right SPL had significantly higher Nv than left SPL and the right PEC. CONCLUSION: Generally, there is an inverse relationship of Nv between the medial and lateral areas of the human posterior parietal cortex. The obtained values were different between the brains with larger left and right hemispheres, as well as between the SPL and PEC. In all the comparisons the left SPL had the highest values of Nv of pyramidal neurons in the layer V (4771.80 mm(-3)), except in brains with the larger right hemisphere.

[Cytochemical and immunocytochemical characteristics of Meckel's diverticulum with heterotopic rests of pancreatic tissue].

BACKGROUND: Meckel's diverticulum (MD) is a congenital anomaly of the small intestine. It results from incomplete obliteration and resorption of the proximal omphaloenteric duct connecting yolk sac with primitive gut in the fetal period. CASE REPORT: A case of 20-year old female with ectopic pancreatic rests in a MD was reported. She was hospitalized with clinical signs of acute appendicitis. During surgery an inflamated Meckel's diverticulum was found and a clinoid resection of the diverticulum was performed. Histologic examination revealed pancreatic tissue in the removed diverticulum. Endocrine cells (EC) were detected with Masson staining and aberrant pancreatic tissue with immunocytochemical LSAB2 method using pan cytokeratin as epithelial marker. CONCLUSION: Most of MD are asymptomatic and accessory finding during laparothomias for different causes, but complications of undiagnozed MD can be serious (diverticulitis, perforation with peritonitis or intestinal obstruction caused by invagination). In unclear cases, additional cytochemical and immunocytochemical diagnostics could be done.

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

BACKGROUND: Two 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. METHODS: Longitudinal 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. RESULTS: Lateral 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. CONCLUSIONS: Although 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.

[Patterns of lipofuscin accumulation in ganglionic nerve cells of superior cervical ganglion in humans].

BACKGROUND/AIM: Considering available literature lipofuscin is a classical age pigment of postmitotic cells, and a consistently recognized phenomenon in humans and animals. Lipofuscin accumulation is characteristic for nerve cells that are postmitotic. This research was focused on lipofuscin accumulation in ganglionic cells (GC) (postganglionic sympathetic cell bodies) of superior cervical ganglion in humans during ageing. METHODS: We analysed 30 ganglions from cadavers ranging from 20 to over 80 years of age. As material the tissue samples were used from the middle portion of the ganglion, which was separated from the surrounding tissue by the method of macrodissection. The tissue samples were routinely fixed in 10% neutral formalin and embedded in paraffin for classical histological analysis, then three consecutive (successive) sections 5 microm thick were made and stained with hematoxylin and eosin method (HE), silver impregnation technique by Masson Fontana and trichrome stain by Florantin. RESULTS: Immersion microscopy was used to analyse patterns of lipofuscin accumulation during ageing making possible to distinguish diffuse type (lipofuscin granules were irregularly distributed and non-confluent), unipolar type (lipofuscin granules were grouped at the end of the cell), bipolar type (lipofuscin granules were concentrated at the two opposite ends of a cell with the nucleus in between at the center of a cell), annular type (lipofuscin granules were in the shape of a complete or incomplete ring around the nucleus) and a cell completely filled with lipofuscin (two subtypes distinguishing, one with visible a nucleus, and the other with invisible one). Even at the age of 20 there were cells with lipofuscin granules accumulated in diffuse way, but in smaller numbers; the GC without lipofuscin were dominant. Growing older, especially above 60 years, all of the above mentioned patterns of lipofuscin accumulation were present with the evident increase in cells completely filled with lipofuscin, but cells without lipofuscin were also present even in the oldest persons. CONCLUSION: Lipofuscin is present in all periods of ageing with a different intensity of accumulation. GC without the pigment, diffusely distributed, as well as very rare cells with a unipolar type of lipofuscin distribution are characteristic for the age of 20-60 years. In the age above 60 years, except the cells without pigment and diffuse accumulation type, there are also bipolar and annular types and forms in which cells are completely filled with lipofuscin granules.

[Anatomohistological characteristics of Meckel's diverticulum in human fetuses].

BACKGROUND/AIM: Meckel's diverticulum (MD) is the most frequent anomaly of the small intestine. It appears after incomplete obliteration of the omphalomesenteric or viteline duct which normaly obliterates and disappears by the 9th week of gestation. The majority of MD do not give rise to any clinical symptoms and are encounted either incidentally, at examination or intervention, or due to complications which may occur (obstruction, hemorrhagy, rupture), and are described in many clinical reports. The aim of the study was to find out the incidence of MD in fetuses when the development of the alimentary tract is already finished. METHODS: The investigation was performed on 150 human fetuses of different sex and gastational age, using microdissection method. The cases with MD were photographed, described, their positions and dimensions were registered. The samples of MD taken for histological investigation were dyed with hematoksilin eosin method. RESULTS: Meckel's diverticulum was found in five fatuses (three male and two female); in one case the fibrous band was found. All of them were located on animesenteric margine of the small intestine at the average distance of 92.5 mm from the ileocecal junction. They were of different shape and dimensions, but of the normal constitution of the small intestine. CONCLUSION: The incidence of MD was 3.3%, and 4% of all the anomalies of the intestines connected to the disappearance of the viteline duct. It was more frequent in the male, located on antimesenteric margine of the small intestine, at the destination which highly correlated to the age of the fetus. Meckel's diverticule were of different shapes and dimensions but of the typical constitution of the small intestine.

Hemodialysis treatment in patients with Balkan endemic nephropathy: an epidemiological study.

AIM: To analyze hemodialysis (HD) treatment of patients with Balkan endemic nephropathy (BEN) from five endemic villages in the South Morava Region of Serbia. Analyses of patterns of incidence may generate hypotheses about the underlying causes of BEN, and prevalence data provide information on the current and likely future burden on health services for managing BEN. METHODS: A total of 143 end-stage kidney disease patients (ESKD) with BEN were admitted to the renal replacement program from 1974 to 2004: 121 to HD, 15 peritoneal dialysis, and 7 kidney transplantation. As a control group, 117 patients with other kidney disease (chronic pyelonephritis, glomerulonephritis, and ischemic nephropathy) admitted to HD at the time of BEN patients and matched by age and gender were studied. RESULTS: Most of the BEN patients (93.4%) treated by HD were born from 1917 to 1941. The majority of patients (79.3%) started HD from 1977 to 1991 (period of 15 years). The mean age of BEN patients starting HD treatment was 49.1 years in the period from 1974 to 1978, and increased steadily in the following years, being 72.5 years in the last period of study (2004-2006) The mean survival time of BEN males was 4.70 (95% CI 3.66-5.75) and for females was 5.02 (95% CI 1.47-4.53). Difference between males and females was not statistically significant (log rank 0.14, p = 0.7, P > 0.5). Mean survival times of 4.84 (95% CI 3.97-5.70) in BEN patients and 3.1 (95% CI 2.78-3.84) in other kidney disease patients were found. Difference between BEN patients and controls was statistically significant (log rank 8.38, p = 0.0038, P < 0.01). CONCLUSION: The population of endemic villages around the South Morava River admitted to HD treatment after 1974 was exposed to environmental toxicant(s) from 1917 to 1941. The most intense effect of environmental exposure was in that period, with ESKD in patients in their forties. The exposure to environmental toxicants has diminished, so ESKD of BEN has become less frequent and manifested in the older age, mean 72.5 in the period from 2004 to 2006. Different type of exposure was registered in some other endemic regions in Serbia and abroad.

Morphometric and some immunohistochemical characteristics of human choroids plexus stroma and psammoma bodies.

Psammoma bodies (PBs) are one of many choroids plexus aging changes. The aim of our research was to perform the quantification of PBs' presence in human choroids plexus stroma, as well as to evaluate the characteristics of choroids plexus stroma in cases in which PBs were present. Afterwards, the observations of the histochemical analysis would be confirmed by immunohistochemical analysis. Choroid plexuses of 30 cadavers were used for the histochemical and, choroids plexuses of 15 cadavers in which PBs' presence was confirmed during the histochemical analysis, were used as material for the immunohistochemical analysis. Light microscopy, histochemical, immunohistochemical, and morphometric method were applied during the study. Classification of the cases was performed by cluster analysis. We observed increase of choroids plexus PBs' presence during the aging process. But this increase is not linear. Their presence is the largest in the second cluster that is younger than the third and older than the first. Nuclear morphometric parameters of the stroma in these cases showed that the cellular composition in this cluster is different than in other two and, that contain larger number of lymphoid cells. Immunohistochemical analysis showed PBs' positive reaction on vimentin, CD45R0, and LCA markers, while in their vicinity, as well as inside them, numerous T-cells were observed. So, the presence of CD45R0 and LCA-positive T cells, PBs' positive reaction on the same markers, indirectly connect these cells with PBs' formation process.

Progression of kidney damage in Balkan endemic nephropathy: a 15-year follow-up of patients with kidney biopsy examination.

Progression of kidney damage was studied in 18 patients with Balkan endemic nephropathy (BEN), with a mean 15-year follow-up after renal biopsy. According to kidney function, estimated by 99mTc-DTPA clearance, patients were divided into three groups: with apparently normal kidney function (clearance 103.5+/-21.3 mL/min/1.73 m2), with incipient renal failure (clearance 65.5 +/- 11.3), and with advanced renal failure (clearance 28.0+/-6.2). The mean yearly decrease of glomerular filtration rate was 2.74 mL/min. In two patients, an increase of kidney function was recorded. Six patients become dialysis dependent, two from the group with incipient renal failure, but all four from the group with advanced renal failure. Three patients died after 8 to 12 years of follow-up, one from causes unrelated to kidney disease and two from end-stage renal failure. This study has shown that BEN is characterized by a slow course and prolonged evolution, modified by medical supervision and treatment.

Morphological and morphometric characteristics of choroid plexus psammoma bodies during the human aging.

Psammoma bodies are one of many choroid plexus aging changes which origin is still enigma for the scientists. During our investigation psammoma bodies were studied on 30 postmortem brains by light microscopy. They stained red with HE, and were PAS and AB PAS positive. The largest number of lamellas were stained blue with Mallory's connective tissue stain, except peripheral and next to the center lamella which stained red. During the aging, psammoma bodies became larger and more irregular, which was followed with group area and perimeter, single psammoma body average area and average perimeter, average diameter and contour index increase. Psammoma bodies mearged in the second and the third age group and mearging process led to larger and more irregular structures formation. The results of this investigation suggest that psammoma bodies are more frequent in choroid plexus of healthy older people and during the aging they obtain larger dimensions, more irregular contours, which is the result of their mutual mearging.