Combustion-Derived Pollutants Linked with Kidney Disease in Low-Lying Flood-Affected Areas in the Balkans.

Tens of thousands of people in southern Europe suffer from Balkan endemic nephropathy (BEN), and four times as many are at risk. Incidental ingestion of aristolochic acids (AAs), stemming from the ubiquitousAristolochia clematitis(birthwort) weed in the region, leads to DNA adduct-induced toxicity in kidney cells, the primary cause of BEN. Numerous cofactors, including toxic organics and metals, have been investigated, but all have shown small contributions to the overall BEN relative to non-BEN village distribution gradients. Here, we reveal that combustion-derived pollutants from wood and coal burning in Serbia also contaminate arable soil and test as plausible causative factors of BEN. Using a GC-MS screening method, biomass-burning-derived furfural and coal-burning-derived medium-chain alkanes were detected in soil samples from BEN endemic areas levels at up to 63-times and 14-times higher, respectively, than in nonendemic areas. Significantly higher amounts were also detected in colocated wheat grains. Coexposure studies with cultured kidney cells showed that these pollutants enhance DNA adduct formation by AA, - the cause of AA nephrotoxicity and carcinogenicity. With the coincidence of birthwort-derived AAs and the widespread practice of biomass and coal burning for household cooking and heating purposes and agricultural burning in rural low-lying flood-affected areas in the Balkans, these results implicate combustion-derived pollutants in promoting the development of BEN.

Hypoplastic arteries of the cerebral arterial ring in the blind spot of computed tomography angiography.

BACKGROUND: Some variations of the cerebral arterial circle (CAC) are associated with an increased risk for the development of various pathological conditions. This paper aimed to determine the prevalence of hypoplastic arteries of CAC and to emphasize the limited possibility of their visualization by computed tomography angiography (CTA). MATERIALS AND METHODS: The research was performed on 400 adult cadavers by macro- and microdissection of the cerebral arteries. Each case was photographed and the diameter of the arteries was measured digitally, by analyzing photographs of the bases of the brain in the ImageJ program. RESULTS: The largest prevalence of artery diameter <1mm (<0.6mm) in CAC had the posterior communicating artery (PCoA). PCoA on the left side was hypoplastic in 44.9% (11.4%) of cases, while the same artery on the right side was hypoplastic in 44.3% (6.6%) of cases. The posterior cerebral artery was hypoplastic on the left side in 3% (0.6%) and on the right side in 4.2% (0.6%) of cases. The anterior cerebral artery had a hypoplastic caliber only on the right side in 2.4% (0.6%) of the cases, while the internal carotid arteries did not have a diameter <1mm in any case. The anterior communicating artery showed the greatest variability in morphology. Studies on CTA describe the occurrence of aplasia in a statistically significantly higher percentage, and the occurrence of hypoplastic arteries in a statistically significantly lower percentage compared to studies on cadavers. CONCLUSIONS: Due to significant differences between cadaveric and radiological studies, it is necessary to analyze their results regarding arterial hypoplasia and aplasia separately. A diameter of less than 1 mm has been suggested as a criterion for arterial hypoplasia.

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 kidney's 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.

Human anterior pituitary's ACTH cells during the aging process: immunohistochemic and morphometric study.

Corticotrophs produce a hormone that stimulates the adrenal gland cortex to secrete glucocorticoids, which in turn have effects on carbohydrate and protein metabolism. Quantification, morphological characteristics, and distribution of corticotrophs in the anterior pituitary and changes in the number and shape of the cells during aging have been examined using immunohistochemical and morphometric methods. The material consisted of 14 anterior pituitaries taken from cadavers at routine autopsy. The tissue was processed by standard histological procedure and the obtained slices were stained by the monoclonal anti-ACTH antibody for corticotrophs identification. Digital images of stained histological sections were analyzed using the morphometric method with the Image J system. The volume density of ACTH positive cells was determined. The cases were classified into three age groups. One-way ANOVA showed that the volume density of the corticotrophs was significantly higher in the second and third group in relation to the first group. The difference in the volume densities of the corticotrophs between the genders was not significant. Morphometric and statistical analyses demonstrated a significant positive correlation between the corticotrophs volume densities and the age of the evaluated cases. Linear regression showed that age significantly predicts corticotrophs volume density. Corticotrophs significantly increase during the life span.

Morphometric analysis of the epineurial and endoneurial blood vessels of the human sciatic nerve in relation to aging.

The aim of this research is to perform an analysis of the epineurial and endoneurial blood vessels in relation to aging. The research is conducted on samples of the human sciatic nerve of 12 case (age from 27 to 89). The histological sections are stained by streptavidin-biotin method of detecting the presence of Type IV collagen. After morphometric analysis the following stereological parameters have been calculated: the number of blood vessels per unit of area, the volume density of the blood vessels and the surface density of the blood vessels of the epineurium and endoneurium. An additional diameter measurement is performed for the endoneural blood vessels. In order to perform a more detailed analysis, the cases were classified into three age groups, the first (27-48 years), the second (49-70 years) and, the third (over 70 years). The bivariate correlation analysis showed that the number of blood vessels of the endoneurium, their volume and surface densities in relation to age produced a statistically significant positive correlation. One Way ANOVA test demonstrated a statistically significant increase in the number of endoneurial blood vessels in the age group III when compared the age group I and, in addition, it showed a significant decrease in the diameter of the age group II when compared to the age group I. Paired t - test shows a statistically significant higher number of endoneurial blood vessels in relation to the epineurial, namely, in the age group III. The volume and surface density of the epineurial blood vessels is significantly higher than the endoneurial in both the I and II age group. Age brings about significant changes of the endoneurial vascular network of the sciatic nerve due to the increase in density of the endoneurial blood vessels, their volume and surface densities. Consequently, in the cases older than 70 years, the number of endoneurial blood vessels significantly exceeds the number of epineurial blood vessels.

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.

Long-lasting undetected neurocysticercosis.

Neurocysticercosis is the most common parasitic disease of the nervous system, nevertheless, it can remain undetected for a long period of time, especially if it occurs in non endemic areas and regions with low-endemicity. Inadequate diagnostic procedures and lack of clinician's dedication towards this health issue can lead to a missed diagnose. Herein, we present a case of a 51-year-old male, with a missed diagnosis of neurocysticercosis for more than two decades. A history of epilepsy had started twenty-one years earlier and was of unclear etiology. Recently, after neurological worsening and headaches, brain computed tomography and magnet resonance imaging was performed as well as Western blot immunoassay of serum and cerebrospinal fluid, surgery, and pathohistological examination of the extracted cysts. Neurocysticercosis was confirmed. Combined therapy that consisted of albendazole and prednisolone was administered for a period of four weeks. Also, antiepileptic therapy was continued. Both clinical status and brain imaging showed the apparent improvement in the patient's condition. Review of the literature was implemented in the discussion that deals with proper and adequate therapy option and outcome factors in neurocysticercosis patients. Over a long period of time, the majority of patients develop seizures as the most common symptom, which requires the administration of medications. Proper diagnostic procedures and adequate combination of surgery and conservative treatment areessential.

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.

[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.

[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.