Braca Kundalić

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.

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.

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 pituitarys 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 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.

Evaluation of Some Anatomical and Anthropometric Characteristics of the Chest Based on the Analysis of Digital Images of the Anterior Aspect of Trunk in Top Athletes

Evaluation of Some Anatomical and Anthropometric Characteristics of the Chest Based on the Analysis of Digital Images of the Anterior Aspect of Trunk in Top Athletes The aim of this research was to assess the size and shape of the chest in students and top athletes. The research involved 23 first-year students of the Faculty of Sport and Physical Education, and 23 top athletes of the Athletic Federation of Serbia. The digital images of the frontal trunk aspect were made and further analyzed in ImageJ program. The vertical and horizontal distances and as well as the angles were determined: the infrasternal angle and the angle of umbilicus (sides of the angle connect the points on the left and right). Both students and athletes were divided into three height groups (I - 165-174 cm; II - 175-184 cm; III - 185-194 cm). BMI and BI were determined (shoulder width). No statistical differences in height, weight and BMI among the groups of students and top athletes were found, which pointed to the homogeneity of the groups. All the parameters determined, the vertical and horizontal ones, except AAD, were significantly higher in top athletes (p ≤ 0.05) compared to the same parameters obtained in students of all three height groups. Acromial distance increases with height, but not statistically significantly. The above mentioned indicates a significantly better development of the bone-joint-muscle system of the chest in top athletes. The infrasternalni angle correlates with the angle of the umbilicus and it can be used to assess the shape of the chest. In our researches, analysing the individual cases, the presence of normasthenic, asthenic (elongated) and barrel-shaped chest was determined. The program ImageJ is very precise, objective and easily applicable for determining the lenghtwise parameters and angles in anatomic and anthropometric measurements. The method does not require anthropometric equipment, digital images can be made quickly and efficiently. Therefore, we consider it particularly suitable for measurements in childhood and athletes. Procena nekih anatomskih i antropometrijskih karakteristika grudnog koša analizom digitalne slike prednjeg aspekta trupa kod vrhunskih atletičara Cilj istraživanja bio je da se proceni veličina i oblik grudnog koša kod studenata i vrhunskih atletičara. Istraživanja su sprovedena na 23 studenta prve godine Fakulteta sporta i fizičkog vaspitanja i 23 vrhunskih atletičara Atletskog saveza Republike Srbije. Načinjene su digitalne fotografije frontalnog aspekta trupa, koje su analizirane u ImageJ programu. Određivane su vertikalne i horizontalne distance i uglovi: infrasternalni i ugao sa temenom kod umbilikusa (kraci ugla spajaju akromijalne tačke levo i desno). I studenti i atletičari podeljeni su u tri visinske grupe (I: 165-174cm; II: 175-184cm; III: 185-194 cm). Određivan je BMI i BI (širina ramena). Nema statističkih razlika u visini, težini i BMI između grupa kod studenata i kod vrhunskih atletičara, što ukazuje na homogenost grupa. Svi određivani parametri, vertikalni i horizontalni, osim AAD, su kod vrhunskih atletičara statistički značajno veći (p 0,05) u odnosu na iste parametre kod studenata u sve tri visinske grupe. Akromijalno-akromijalna distanca se povećava sa visinom, ali ne statistički značajno. Napred navedeno ukazuje na značajno bolju razvijenost koštanozglobno-mišićnog sistema grudnog koša kod vrhunskih atletičara. Infrasternalni ugao korelira sa uglom kod umbilikusa, te se može koristiti za procenu oblika grudnog koša. U našim istraživanjima, analizom pojedinačnih slučajeva, utvrđeno je prisustvo normosteničnog, asteničnog i bačvastog oblika grudnog koša. Program ImageJ je veoma precizan, objektivan i lako primenljiv za određivanje dužinskih parametara i uglova u anatomskim i antropometrijskim merenjima. Metoda ne zahteva antropometrijsku opremu, a digitalne slike se mogu brzo i efikasno uraditi, te smatramo da je posebno pogodna kod merenja u dečijem uzrastu i kod sportista.