Dilated Cisternae Research Articles

Effect of acute trihexyphenidyl abuse on rat motor area of cerebral cortex

Introduction Trihexyphenidyl (THP) is an antiparkinsonian drug, most frequently abused to achieve pleasurable effects. In contrast, the effect of THP abuse on the motor area of the frontal cortex has not been adequately investigated. Aim of the work To study the histological changes of different single doses of TPH on rat motor area. Materials and methods Forty-eight adult male rats were used in this study. The rats were divided equally into four groups: a control group (group I), and three experimental groups (groups II, III, and IV) which were intraperitoneally injected with a single variable dose of TPH (0.2, 0.5, 1.0 mg/kg/body weight, respectively). After 6 h of injection, blood samples were collected for measurement of serum THP level. Specimens from the frontal cortex motor area were taken, processed, and examined by light and electron microscopes. Results By microscopic examination, the motor area of group II rat which received a TPH dose equivalent to the human therapeutic dose was nearly similar to that of the control. In group III, TPH induced apoptotic changes in the pyramidal and granular cells. Some pyramidal cells exhibited ultrastructural changes including; dilated Golgi saccules, dilated rough endoplasmic reticulum cisternae, multiple lysosomes, and cristolysis of the mitochondria. Moreover, the axons exhibited axoplasmic vacuoles and multiple splitting of the myelin lamellae. Group IV motor areas, showed multiple apoptotic cells and focal necrotic areas with microglial infiltration. Electron microscopic study revealed degenerated pyramidal cells and extravasation of red blood corpuscles. The axons showed multiple focal losses of the myelin lamellae. Conclusion TPH induced dose-dependent structural changes in rat frontal cortex motor area. Therefore, THP prescription must be under strict supervision.

Neuroprotective effects of Caffeic acid phenethyl ester on experimental traumatic brain injury in rats

The aim of this study was to evaluate the therapeutic efficacy of caffeicacid phenethyl ester (CAPE) with an experimental traumatic brain injury (TBI) model in rats. Twenty-four adult male Sprague-Dawley rats were randomly divided into three groups of 8 rats each: control, TBI, and TBI+CAPE treatment. In TBI and TBI+CAPE treatment groups, a cranial impact was delivered to the skull from a height of 7cm at a point just in front of the coronal suture and over the right hemisphere. Rats were sacrificed at 4h after the onset of injury. Brain tissues were removed for biochemical and histopathological investigation. To date, no biochemical and histopathological changes of neurodegeneration in the frontal cortex after TBI in rats by CAPE treatment have been reported. The TBI significantly increased tissue malondialdehyde (MDA) levels, and significantly decreased tissue superoxide dismutase (SOD) and glutathione peroxidase (GPx) activities, but not tissue catalase (CAT) activity, when compared with controls. The administration of a single dose of CAPE (10μmol/kg) 15min after the trauma has shown protective effect via decreasing significantly the elevated MDA levels and also significantly increasing the reduced antioxidant enzyme (SOD and GPx) activities, except CAT activity. In the TBI group, severe degenerative changes, shrunken cytoplasma and extensively dark picnotic nuclei in neurons, as well as vacuolization indicating tissue edema formation. The morphology of neurons in the CAPE treatment group was well protected. The number of neurons in the trauma alone group was significantly less than that of both the control and TBI +CAPE treatment groups. The caspase 3 immunopositivity was increased in degenerating neurons of the traumatic brain tissue. Treatment of CAPE markedly reduced the immunoreactivity of degenerating neurons. TBI caused severe degenerative changes, shrunken cytoplasma, severely dilated cisternae of endoplasmic reticulum, markedly swollen mitochondria with degenerated cristae and nuclear membrane breakdown with chromatin disorganization in neurons of the frontal cortex. In conclusion, the CAPE treatment might be beneficial in preventing trauma-induced oxidative brain tissue damage, thus showing potential for clinical implications. We believe that further preclinical research into the utility of CAPE may indicate its usefulness as a potential treatment on neurodegeneration after TBI in rats.

Synthesis of keyhole limpet hemocyanin by the rhogocytes ofMegathura crenulata

Rhogocytes are morphologically distinct cells distributed throughout connective tissues of crustaceans and molluscs. Using light microscopy, rhogocytes of the vetigastropod Megathura crenulata were identified by their ovoid shape, and their cytoplasm filled with spherical inclusions which contained lysosomal enzymes, based on uptake of neutral red and staining with LysoTracker dye. Rhogocytes were most abundant in the digestive gland (2,824 rhogocytes/mm^2), followed by the connective tissue layer surrounding the middle and posterior esophagus and intestine (1,431 rhogocytes/mm^2, 872 rhogocytes/mm^2, and 1,190 rhogocytes/mm^2, respectively), and were lowest in abundance in the foot (154 rhogocytes/mm^2). At the transmission electron microscopy level, characteristic features of rhogocytes were inclusions showing a variety of electron densities, abundant vesicles, and rough endoplasmic reticulum in the cytoplasm, and regions of plasma membrane folded to produce slits connected by thin diaphragms. Although several functions have been proposed for gastropod rhogocytes, much attention has been focused on their possible role in the synthesis of the respiratory pigment hemocyanin. In M. crenulata, this molecule exists in several isoforms called keyhole limpet hemocyanin (KLH). One isoform, KLH1, is a large didecamer and has been used extensively in studies on vertebrate immunology and cancer therapy. We present four lines of evidence indicating rhogocytes in M. crenulata synthesize KLH1. First, at the transmission electron microscopy (TEM) level, dilated cisternae of RER containing material similar in size and shape to KLH were observed in rhogocytes examined throughout the year. Second, KLH1 mRNA was identified exclusively in tissue samples that contained rhogocytes; no mRNA for KLH1 was identified in samples containing only hemocytes. Third, immunoperoxidase staining with antibodies specific to KLH was localized only to rhogocytes. Fourth, in situ hybridization with a probe specific for M. crenulataKLH1 demonstrated KLH1-specific mRNA was present only in rhogocytes. Identification of the cells responsible for the synthesis of KLH is important because of the clinical significance of this molecule.

Biochemical and ultrastructural study of the effects of proline analogues on collagen synthesis in 3T6 fibroblasts.

The effects of proline analogues, L-3,4-dehydroproline and L-azetidine-2-carboxylic acid, on collagen synthesis by cultured 3T6 fibroblasts have been studied. Prolyl hydroxylase activity was partially inhibited in cells cultured with dehydroproline for 24 h, resulting in the synthesis of collagen in which the proline was underhydroxylated. Azetidine had no effect on prolyl hydroxylase and less effect on the degree of hydroxylation of proline. Fibroblasts grown in the presence of either analogue and fixed in-situ contained greatly distended cisternae of the rough endoplasmic reticulum. Proline analogues otherwise caused few ultrastructural changes in the cells. Treated cells which had been handled more roughly during preparation for electron microscopy contained many large cytoplasmic vacuoles in addition to dilated cisternae. Our results indicate that the major effect of the proline analogues was the inhibition of prolyl hydroxylation. However, electron microscopy of the treated cells revealed hitherto unreported cytoplasmic damage.

Cells producing insulin-like androgenic gland hormone of the giant freshwater prawn, Macrobrachium rosenbergii, proliferate following bilateral eyestalk-ablation

We found that the androgenic gland (AG) of Macrobrachium rosenbergii possesses three cell types. Type I cells are small polygonal shaped-cells (13.4 μm in diameter), stain strongly with hematoxylin–eosin (H&E), have abundant multilayered rough endoplasmic reticulum (rER), and nuclei containing mostly heterochromatin. Type II cells are slightly larger (18.6 μm in diameter), stain lightly with H&E, have rER with dilated cisternae, and nuclei containing mostly euchromatin. Type III cells (previously undescribed) are similar in size and shape to type I cells, but the cytoplasm is unstained and they have a high amount of smooth endoplasmic reticulum (sER) and mitochondria with tubular cristae. Bilateral eyestalk-ablation resulted in AG hypertrophy with a proliferation and predominance of type I cells as determined by bromodeoxyuridine (BrdU) assays. Expression of insulin-like androgenic gland hormone (Mr-IAG), determined by immunohistochemistry, was weak in type I cells, strong in type II cells of both the intact and eyestalk-ablated, and negative in type III cells. It was also detected in spermatogonia, nurse cells, and epithelium lining of the spermatic duct. The function of Mr-IAG in these tissues is yet to be elucidated but the distribution implies a strong role in male reproduction.

A comprehensive characterization study of human bone marrow mscs with an emphasis on molecular and ultrastructural properties

Human bone marrow-derived mesenchymal stem cells (hBM-MSCs) continue to draw attention of researchers in the fields of basic science and medicine due to their indispensible regenerative, reparative, angiogenic, anti-apoptotic, and immunosuppressive properties, all of which collectively point out their enormous therapeutic potential. There is still, however, a need for further investigation of their characteristics to broaden their field of use and learn much more about how to control their fate and improve their therapeutic effectiveness. hBM-MSCs were extensively characterized in terms of their growth characteristics, genetic stability, and differentiation capability to the mesodermal and ectodermal cell lineages; a special emphasis was given to their phenotypic and ultrastructural properties. Expression of embryonic stem cell markers Oct4, Rex-1, FoxD-3, Sox2, and Nanog was shown with real-time PCR. Transmission electron microscopy revealed the ultrastructural characteristics of hBM-MSCs; they had pale, irregularly shaped and large euchromatic nuclei, and two distinct areas in their cytoplasm: an intensely stained inner zone rich in mitochondria and rough endoplasmic reticulum (rER) with dilated cisternae and a relatively peripheral zone poor in organelles. hBM-MSCs expressed adipogenic (adipophilin and PPARγ), myogenic (desmin, myogenin, α-SMA), neurogenic (γ-enolase, MAP2a,b, c-fos, nestin, NF-H, NF-L, GFAP, β3-tubulin), osteogenic (osteonectin, osteocalcin, osteopontin, Runx-2, type I collagen), and chondrogenic (type II collagen, SOX9) markers either at RNA or protein level even under basal conditions, without any stimulation towards differentiation. The differentiation potential of hBM-MSCs to adipogenic, osteogenic, and neurogenic lineages was shown by using the relevant differentiation factors.

Direct imaging of pH1N1 2009 influenza virus replication in alveolar pneumocytes in fatal cases by transmission electron microscopy

Human influenza virus pandemics constitute a major global public health issue. Although studies on autopsy specimens from the recent pandemic by the 2009 influenza A (H1N1) virus have revealed a broad spectrum of pathologic findings, direct electron microscopic studies of the lung tissue from influenza fatalities are few. In this study, we examined five well-preserved pulmonary necropsy specimens from fatal cases of laboratory-confirmed pH1N1 from India. The novel observations in comparison with earlier reports included direct imaging of influenza virus budding within dilated cisternae of pneumocytes, cell-free virus emerging from the cell membrane of a pneumocyte in the alveolar lumen, presence of polymorphonuclear cells with red blood cells as inflammatory exudates close to hyaline membranes and extensive cytoplasmic degeneration of epithelial cells of the alveolar lining. These observations are in consistent with the earlier findings and emphasize the possible role of this virus directly infecting cells of the lower respiratory tract as a key event in the rapid pathogenesis of pH1N1 disease process.

Inflammation, russell bodies, GRP78 and endoplasmis reticulum stress – are they linked?

Introduction GRP78 has a critical role in the attenuation of cellular stress and recovery from injury. It is involved in the folding and assembly of proteins in the endoplasmic reticulum (ER) and is induced in times of ER stress. Russell bodies (RB) are dilated ER cisternae containing condensed immunoglobulins and are thought to arise from failure of ER quality control mechanisms. Purpose To investigate the relationship between Russell bodies and ER stress in periodontal inflammation. Methods Formalin fixed paraffin embedded (FFPE) specimens were retrieved and categorised into three groups: inflamed period-ontal tissues with RB (Group 1), inflamed tissues without RB (Group 2), and lightly inflamed control tissue (Group 3). Serial sections were prepared and stained with H&E, PAS, methyl green pyronin and anti-GRP78. The number of RB and the distribution and types of cells staining positively with GRP78 were analysed. Results Of the inflammatory cells in Groups 1 and 2 which were GRP78+, most were plasma cells. In the less inflamed Group 3 tissues lymphocytes were also GRP78+. Inflamed tissues had statistically significant more apoptotic/degenerate bodies than control tissues. There was no statistically significant difference between GRP78 staining and the presence or absence of RB in inflamed tissues and control tissues. Conclusion Inflammation, rather than RB, correlated positively with GRP78 immunostaining.

Dedifferentiation of prostate smooth muscle cells in response to bacterial LPS

Prostate smooth muscle cells (SMCs) are strongly involved in the development and progression of benign prostatic hyperplasia and prostate cancer. However, their participation in prostatitis has not been completely elucidated. Thus, we aimed to characterize the response of normal SMC to bacterial lipopolysaccharide (LPS). Primary prostate SMCs from normal rats were stimulated with LPS (0.1, 1, or 10 µg/ml) for 24 or 48 hr. The phenotype was evaluated by electron microscopy, immunofluorescence, and Western blot of SMCα-actin (ACTA2), calponin, vimentin, and tenascin-C, while the innate immune response was assessed by immunodetection of TLR4, CD14, and nuclear NF-κB. The secretion of TNFα and IL6 was determined using ELISA. Bacterial LPS induces SMCs to develop a secretory phenotype including dilated rough endoplasmic reticulum cisternae with well-developed Golgi complexes. Furthermore, SMCs displayed a decrease in ACTA2 and calponin, and an increase in vimentin levels after LPS challenge. The co-expression of ACTA2 and vimentin, together with the induction of tenascin-C expression indicate that a myofibroblastic-like phenotype was induced by the endotoxin. Moreover, LPS elicited a TLR4 increase, with a peak in NF-κB activation occurring after 10 min of treatment. Finally, LPS stimulated the secretion of IL6 and TNFα. Prostate SMCs are capable of responding to LPS in vitro by dedifferentiating from a contractile to a miofibroblastic-like phenotype and secreting cytokines, with the TLR4 signaling pathway being involved in this response. In this way, prostate SMCs may contribute to the pathophysiology of inflammatory diseases by modifying the epithelial-stromal interactions.

Protective effects of thymoquinone on the neuronal injury in frontal cortex after chronic toluene exposure

The aim of this study was designed to evaluate the possible protective effects of thymoquinone (TQ) on the neuronal injury in the frontal cortex after chronic toluene exposure in rats. The rats were randomly allotted into one of three experimental groups: A (control), B (toluene treated) and C (toluene treated with TQ); each group contain 10 animals. Control group received 1 ml normal saline solution and toluene treatment was performed by inhalation of 3,000 ppm toluene, in a 8 h/day and 6 day/week order for 12 weeks. The rats in TQ treated group was given TQ (50 mg/kg body weight) once a day orally for 12 weeks starting just after toluene exposure. Tissue samples were obtained for histopathological investigation. To date, no histopathological changes of neurodegeneration in the frontal cortex after chronic toluene exposure in rats by TQ treatment have been reported. In this study, the morphology of neurons in the TQ treatment group was well protected. Chronic toluene exposure caused severe degenerative changes, shrunken cytoplasma, severely dilated cisternae of endoplasmic reticulum, markedly swollen mitochondria with degenerated cristae and nuclear membrane breakdown with chromatin disorganization in neurons of the frontal cortex. We conclude that TQ therapy causes morphologic improvement on neurodegeneration in frontal cortex after chronic toluene exposure in rats. We believe that further preclinical research into the utility of TQ may indicate its usefulness as a potential treatment on neurodegeneration after chronic toluene exposure in rats.

Chondroid chordoma of the skull base: immunohistochemical and ultrastructural study of two cases with special reference to microtubules within rough-surfaced endoplasmic reticulum

Two cases of skull base chordoma (case 1, a 57-year-old woman; case 2, a 69-year-old woman) were investigated immunohistochemically and ultrastructurally. The tumors showed histopathological features typical of chondroid chordoma and contained both classical chordomatous and hyaline cartilaginous components. Tumor cells were immunoreactive for cytokeratin, vimentin, and S-100 protein, but negative for microtubule-associated protein 2 and class III beta-tubulin (tub-B3). Tumor cells of case 2 were immunoreactive for tau-protein and class II beta-tubulin (tub-B2), whereas those of case 1 were negative. Ultrastructurally, tumor cells in both cases showed the presence of abundant glycogen granules, well-developed intracellular organelles, and desmosome-like junctions. In case 2, several microtubules were closely packed and ran parallel or in random directions within the dilated cisterns of rough-surfaced endoplasmic reticulum (rough ER). "Microtubules within rough ER" has been described in several neoplasms, including classical and chondroid chordomas. Although previous reports documented the tub-B3 immunoreactivity in chordomas, our results suggested that, in our case 2, the predominant isoform of beta-tubulin in microtubules within rough ER was not tub-B3 but tub-B2.

Russell body gastritis

Russell body gastritis, first described in 1998 [1], is a reactive gastric mucosal infiltration of plasma cells filled with cytoplasmic Russell bodies. Russell bodies are immunoglobulins present within dilated rough endoplasmic reticulum cisternae that have accumulated secondary to abnormal secretion [2]. We present a case of Russell body gastritis occurring in a 75-year-old man with a history of alcohol use, renal failure, dyslipidemia, and prior rhabdomyolysis. He was noncompliant with reflux medications and reported intermittent coffee-ground emesis. Esophagogastroduodenoscopy with biopsy revealed esophagitis and nodular chronic active gastritis. The gastric antral mucosa exhibited regenerative changes and a dense chronic inflammatory infiltrate (Image 1) composed of numerous Russell body filled plasma cells, including Mott cells (Image 2). No lymphoepithelial lesions were identified. A CD138 immunostain (Image 3) confirmed that these were plasma cells and in situ hybridization studies for kappa and lambda light chains demonstrated their polyclonal distribution. Gastric biopsies were negative for Helicobacter pylori by immunohistochemistry. Regenerative gastric mucosa showing numerous plasma cells filled with bright eosinophilic Russell bodies (H&E stain; original magnification ×200). Plasma cells shown at higher magnification are filled with cytoplasmic Russell bodies (H&E stain; original magnification ×600). Plasma cells show strong CD138 immunoreactivity (immunohistochemical stain; original magnification ×600). To date, there have been 11 other published case reports of Russell body gastritis [1-10]. The average patient age is 57 years (range, 29–80 years), including seven males and five female patients. These patients presented primarily with gastric related symptoms including nausea, dyspepsia, and epigastric pain. Mucosal swelling and erythema appear to be relatively common endoscopic findings, and only a few cases exhibited erosion, ulceration, or a raised lesion endoscopically. The exact etiology of Russell body gastritis is unclear. However, nine (82%) of these reported cases were H.pylori positive, two individuals were positive for human immunodeficiency virus (HIV), and one also had Candida esophagitis. Another two patients had Epstein Barr virus (EBV)-associated gastric carcinoma. Except for one individual with a concomitant monoclonal gammopathy of undetermined significance (MGUS), none of these patients have manifested a plasma cell disorder. Russell body gastritis is a diagnosis based on microscopic findings. The differential diagnosis on biopsy includes signet ring cell carcinoma, plasmacytoma, B-cell lymphoma with plasmacytic differentiation, and granular cell tumor. As highlighted in this, ancillary studies can be useful to demonstrate that these cells are polytypic plasma cells. Associated gastric carcinoma and infection (H.pylori, HIV, EBV, Candida) should also be excluded.

Ultrastructural morphology of equine adipose-derived mesenchymal stem cells.

Mesenchymal stem cells are a virtually ubiquitous population of adult stem cells, able to differentiate into various tissue lineages. As they are multipotent and easy to grow in culture, they are at present considered very attractive candidates for tissue repair and gene therapy. With the exception of a few reports, mesenchymal stem cell morphology has been widely disregarded in the past years. In this paper we discuss the establishment of mesenchymal stem cell cultures from equine adipose tissue and describe their fine structure by transmission electron microscopy. The cultured cells revealed a fibroblastoid appearance and were characterized by an eccentric nucleus with multiple nucleoli, dense cytoplasm rich in ribosomes, a rough endoplasmic reticulum with dilated cisternae, elongated mitochondria and heterogeneous vacuolar inclusions. In addition, they were often interconnected by adhesion structures located on the cell body and on cytoplasmic processes contacting other cells. The features observed are evocative of an undifferentiated cellular phenotype and of an intense synthetic and metabolic activity.

Response of IMCD3 cells to hypertonic challenges as analyzed by electron microscopy

This work defines the ultrastructural responses of immortalized cells from the inner medullary collecting duct cells (IMCD3 cells) to hypertonic challenges. The cultured cells were either acutely exposed to hypertonic medium (550 mOsm/kgH₂O) for 24-72 h or gradually adapted to 600 or 900 mOsm/kgH₂O media with sodium chloride. After short (24 h) hypertonic challenges, there was an expansion of the Golgi apparatus with distinct expression of the γ subunit of Na,K-ATPase. The frequency of active caspase-3-positive cells was unchanged as was also the measured activity of caspase-3. Immunoelectron microscopy showed that active caspase-3 in the positive cells was localized in cytoplasmic bodies 0.5-1 μm in diameter but not in other structures. Apoptotic bodies with the nuclei were only rarely observed following acute hypertonicity for 24 to 72 h. Following prolonged hypertonic challenges, some cells showed condensation of the chromatin but still few apoptotic bodies. Gradual hypertonicity to 900 mOsm/kgH₂O led to a decrease of microvilli, dilated cisternae of the endoplasmic reticulum (ER), increased abundance of free ribosomes and longitudinal mitochondrial cristae. Virus particles were present inside and outside the cells in all experimental conditions and appeared unrelated to the apoptotic process. The results suggest that cultured IMCD3 cells are resistant to short hypertonic challenge or gradual adaptation to moderate hypertonicity and only rarely exhibit more ultrastructural apoptotic changes than control cells. The presence of caspase-3-containing bodies is a novel finding, and we suggest that they arise from the ER and are involved in the apoptotic signaling system.

The Possible Protective Role of Vitamin E on Fluoride Induced Neurotoxicity in the Frontal Cortex of Adult Male Albino Rats

Introduction: Sodium fluoride is commonly added to drinking water, tooth pastes and some mouth washes as decay preventive ingredient. However, it becomes toxic at long term exposure and leads to some adverse effects on various organs, including brain. Aim of the Work: Investigate the possible histological and immunohistochemical changes in the frontal cortex of sodium fluoride intoxicated rats and evaluate the possible protective role of vitamin E. Materials and Methods: Thirty-six adult male albino rats were used in this study and divided equally into three groups: control, treated and prophylactic. Treated group comprised animals that received a daily oral dose of sodium fluoride (20mg /kg) for 4 weeks. Prophylactic group received a daily oral dose of sodium fluoride (20 mg/kg) concomitant with vitamin E (400mg/kg) for the same duration. At the end of experiment, the frontal cortex of all rats was processed for light and electron microscopic examination. Immunohistochemical study was performed for detection of Glial Fibrillary Acidic Protein (GFAP) and Bcl-2. The obtained results were analyzed morphometrically and statistically. Results: Light microscopic examination of treated group revealed that most nerve cells in the cortical layers were distorted in shape with deeply stained shrunken nuclei. The ultrastructural study exhibited apoptotic neurons with heterochromatic nuclei. Their cytoplasm contained dilated and fragmented cisternae of rough endoplasmic reticulum. Immunohistohemical study of this group revealed intense positive immunoreaction for GFAP in the cytoplasm and processes of astrocytes while weak reaction for Bcl-2 was observed in most of nerve cells. The prophylactic group displayed normal appearance of most of nerve cells in the cortical layers and few cells appeared distorted. Positive immunoreaction for Bcl-2 was observed in many nerve cells and endothelium of blood vessels. Area percent of GFAP immunoreaction in astrocytes in all groups showed significant increase in the treated group when compared with the control or prophylactic groups. Conclusion: Chronic exposure to sodium fluoride results in histological and immunohistochemical changes on the neurons and glial tissue of frontal cortex. Furthermore, combined vitamin E supplementation has an ameliorating effect on these changes. So, dietary supplements of vitamin E may be beneficial for populations at risk

Morphologic Features and Flow Void Phenomenon in Normal Pressure Hydrocephalus and Other Dementias: Are They Really Significant?

The aim of this study was to determine the distinctive features of normal-pressure hydrocephalus (NPH) and other dementias on routine T1-weighted and T2-weighted magnetic resonance (MR) images. Also, the contribution of these parameters to the diagnosis and treatment of NPH was investigated. Routine MR images were used to investigate the morphologic features (dilatation of Sylvian cisterns, narrowness of convexity sulci, thickness of corpus callosum (TCC), and dilatation of perihippocampal fissures) and the flow void phenomenon (FVP) in patients with idiopathic NPH (INPH) and other dementias. Routine MR images of 18 patients with INPH, 11 with dementias other than INPH, and 20 controls were retrospectively examined. Morphologic features and the FVP were graded subjectively. The TCC was measured quantitatively. Morphologic parameters, the FVP, and the shunt response were assessed using Kruskal-Wallis and Mann-Whitney U tests. The mean FVP score was significantly higher in patients with INPH (2.89 +/- 0.75) than in controls and patients with other dementias (1.1 +/- 0.85 and 1.09 +/- 0.83, respectively) (P < .001). There was significant difference in terms of TCC between patients with INPH (3 +/- 0.7 mm), those with other dementias (1.9 +/- 0.7 mm), and controls (5.2 +/- 0.8 mm) (P < .001). Significant differences in terms of other morphologic features were found between patients with INPH and those with other dementias (P < .05). No significant difference was found between morphologic parameters and the FVP and the outcome of cerebrospinal fluid diversion (P > .05). Intense FVP is a signature of but is not pathognomonic for INPH. The morphologic analysis of MR images can be distinctive for the diagnosis of INPH or dementias other than INPH. Detailed evaluation of morphologic features and the FVP in routine MR workup of dementia will be useful for accurate diagnosis.

Hyperglycaemia impairs antithrombin secretion: Possible contribution to the thrombotic risk of diabetes

Recent data support that diabetes might be a conformational disease. Certainly, hyperglycaemia causes a broad range of deleterious effects that might facilitate protein aggregation. We have evaluated the effects of hyperglycaemia on antithrombin, a conformationally sensitive serpin with a potent anticoagulant role. Moreover, these studies might also help to understand the thrombotic risk associated to diabetes. We incubated in vitro plasma and purified antithrombin and human hepatoma cells (HepG2) with methyl-glyoxal and glucose. Moreover, a mouse model of acute diabetes was generated with streptozotocin. Antigen, anti-FXa activity, heparin affinity and conformational features of antithrombin were analysed. Histological and intracellular features and distribution of antithrombin in HepG2 and livers of mice were also evaluated. Hyperglycaemia i n vitro induced a transition of antithrombin to a form with low heparin affinity that explained the loss of anticoagulant activity, without generation of abnormal conformers (polymers or latent antithrombin). However, these effects were not observed on circulating antithrombin from diabetic mice. In contrast, hyperglycaemia in vivo had significant effects on intracellular antithrombin, which was retained, forming microaggregates within the lumen of dilated cisterns of the endoplasmic reticulum. These effects explained the moderate type I deficiency observed in diabetic mice. Similar intracellular consequences were observed for another hepatic serpin, α1-antitrypsin. Our data further support that diabetes has conformational effects on structurally sensitive proteins. These effects on antithrombin, the main natural anticoagulant, might contribute to the hypercoagulable status of diabetic patients.

Cerebellar cortical degeneration in cattle caused by Solanum kwebense

The pathology of maldronksiekte, a sporadic neurological disorder of cattle caused by the ingestion of the plant Solanum kwebense in certain parts of South Africa, was studied in three chronic field cases. There was loss of cerebellar Purkinje cells with the remaining neurons either swollen or shrunken and showing degeneration and necrosis. Ultrastructurally, neurons with a swollen perikaryon showed depletion and empty dilated cisternae of granular endoplasmic reticulum. In a few Purkinje cells, the cytoplasm contained small numbers of lamellar and membranous bodies. In the shrunken neurons, the highly condensed cytoplasm contained distended Golgi saccules, dense clusters of granular endoplasmic reticulum and swollen mitochondria. Lectin histochemistry revealed that the cytoplasmic vacuoles in some distended Purkinje cells stained strongly with Canavalia ensiformis (ConA) agglutinin and weakly with Triticum vulgaris (WGA) and succinyl-WGA (S-WGA) agglutinin. The pattern of lectin binding only partially agreed with that reported in calves poisoned with Solanum fastigiatum, causing a presumed glycolipid storage disease. Apoptosis was not detected in neurons using a commercial deoxyuridine triphosphate nick-end labelling (TUNEL) method. The pathogenesis of the cerebellar lesions is unknown but the intoxication may have resulted from the inability of neurons, in particular Purkinje cells, to metabolise a plant toxin or cellular substrate.

Toxicidade da mitomicina C ao endotélio da córnea de coelhos

To evaluate corneal endothelium alterations after applying mitomycin C to the sclera using transmission and scanning electron microscopy, correlating alterations with time, concentration, and evaluation methods. The corneal endothelium of both eyes of 32 albino rabbits was evaluated and distributed into four groups of 8. Mitomycin C was applied under a scleral flap in the right eye for 5 minutes. Mitomycin C concentrations were 0.5 mg/ml for G1 and G2 and 0.2 mg/ml for G3 and G4. Examinations were performed 15 days after application to G1 and G3, and 30 days after application to G2 and G4. Four cornea in each group were prepared for transmission electron microscopy and four for scanning electron microscopy. Left eyes of all animals were used as controls. Transmission electron microscopy showed corneal endothelium alterations in all groups: rarefied cytoplasm, dilation and fragmentation of rough endoplasmic reticulum cisternae, Golgi apparatus with cisternal dilation, reduced vacuoles, and irregularities of internal membrane more noticeable in G1 and G2. Scanning electron microscopy revealed alterations in all groups except G1: changes in the shape and size of cells and longer filopodial projections. 1 - Corneal endothelium alterations were seen at both 0.5 and 0.2 mg/ml concentrations and at 15 and 30 days after mytomicin C application; 2 - Alterations were more intense with higher mytomicin C concentration by transmission electron but not by scanning electron microscopy; 3 - The alterations correlated with time by scanning electron microscopy but not by transmission electron microscopy.

Ultrastructural changes of the olfactory bulb in manganesetreated mice

The effect of manganese toxicity on the ultrastructure of the olfactory bulb was evaluated. Male albino mice were injected intraperitoneally with MnCl2 (5 mg/Kg/day) five days per week during nine weeks. The control group received NaCl (0.9%). The olfactory bulbs of five mice from each group were processed for transmission electron microscopy after 2, 4, 6 and 9 weeks of manganese treatment. On week 2, some disorganization of the myelin sheaths was observed. After 4 weeks, degenerated neurons with dilated cisternae of rough endoplasmic reticulum and swollen mitochondria appeared. A certain degree of gliosis with a predominance of astrocytes with swollen mitochondria, disorganization of the endomembrane system, dilation of the perinuclear cisternae and irregularly shaped nuclei with abnormal chromatin distribution were observed after 6 weeks. Some glial cells showed disorganization of the Golgi apparatus. On week 9, an increase in the number of astrocytes, whose mitochondrial cristae were partially or totally erased, and a dilation of the rough endoplasmic reticulum were found. Neurons appear degenerated, with swollen mitochondria and a vacuolated, electron dense cytoplasm. These changes seem to indicate that the olfactory bulb is sensitive to the toxic effects of manganese.