Fenugreek powder exerts protective effects on alcoholised rats' kidney, highlighted using ultrastructural studies.

Trigonella foenum-graecum (TFG) seeds exert a protective antioxidant effect and membrane protector through their rich content in polyphenolic flavonoids. The previous research focused on the hypoglycemic action of the seeds, with scarce studies on the preventive effects in the pathology of the kidney. Our work was conducted on an experimental in vivo model; the animals were given two different concentrations of TFG seeds, consequently to alcohol intoxication. Transmission electron microscopy (TEM) analysis showed vacuolation in cytoplasm, edemas at the apical pole of the nephrocytes, diffusion of the cytoplasmic and mitochondrial matrix and the increase in number of the lysozymes and especially peroxisomes, as well as the congestion of blood capillaries. In the case of the groups T5R and T10R, which received Trigonella powder together with ethanol, the structural and ultrastructural changes produced by the ethylic intoxication were more reduced, being somewhat improved in the T5R group. Therefore, the majority of the cells nuclei have retained their spherical shape, being at the same time predominantly euchromatic, with little heterochromatin and evenly dispersed. Our results suggest the use of Trigonella seeds as a food supplement to prevent cellular deterioration and improve renal morphology and function.

Fractal analysis of ibuprofen effect on experimental dog peri-implantitis.

INTRODUCTION: The aim of this study was to assess the correlation between the fractal analysis of gingival changes and systemic nitro-oxidative stress in a short-term low-dose ibuprofen (IBU) treatment at experimental peri-implantitis (PI). MATERIALS AND METHODS: Six adult male mixed-breed dogs with PI were randomly treated for 2 weeks, 3 with IBU (5 mg/kg b.w.) and 3 with placebo. Clinical and radiological evaluation were performed. Gingival biopsies were assessed by light microscopy, transmission electron microscopy, and fractal analysis. Blood was collected to assay nitric oxide (NOx), total oxidative status (TOS), total antioxidant response (TAR), and oxidative stress index (OSI). RESULTS: Specific gingival ultrastructural alterations, bone loss, and systemic nitro-oxidative stress were evident in PI-placebo animals. IBU caused significant clinical, microscopic, fractal dimensions (P < 0.01), NOx, TOS, and OSI improvements. IBU caused no important bone and TAR changes. CONCLUSION: This study confirms that fractal analysis was a good method to assess the complex morphological changes and correlations with the nitro-oxidative stress in PI. Short-term low-dose IBU treatment consistently improved gingival status and reduced systemic nitro-oxidative stress.

Porosome in the Exocrine Pancreas: A Detailed EM Study suppressor.

A major question in cell biology that accumulation of partially empty vesicles in cells following secretion is seen, while it is believed that secretion occurs via the complete merger of secretory vesicles with the cell plasma membrane. This important question was solved nearly two decades ago, with the discovery of the Porosome. Porosomes are cup-shaped lipoprotein structures found at the plasma membrane of all cells. Secretory vesicles dock and transiently fuse at the porosome base to form a continuous channel or fusion pore to release the pressurized vesicle contents to the outside. In a decade-long study by our group, we carefully examined using electron microscopy, the detailed structure of the porosome complex in acinar cells of the exocrine pancreas. Besides conformation of earlier findings, our study provides in much greater detail, the in situ morphology of the porosome complex in the exocrine pancreas. The discovery of the detailed morphology of the exocrine pancreas porosome complex in my laboratory is one of the major highlights of my academic career spanning nearly 50 years. Results from our EM studies, reveal for the first time the presence of tethers or cables, which are likely t-SNAREs, present at the porosome base. These EM studies further demonstrate for the first time the docking of a single secretory vesicle or zymogen granule at the base of more than one porosome complex. Detailed spoke-like elements lining the porosome cup were also observed for the first time in these studies, greatly advancing our understanding of the molecular architecture and physiology of the porosome in the exocrine pancreas.

Bee venom induced in vivo ultrastructural reactions of cells involved in the bone marrow erythropoiesis and of circulating red blood cells.

Ultrastructural answer of bone marrow erythroid series and of red blood cells (RBCs) in Wistar rats to bee venom (BV) was analyzed by transmission and scanning electron microscopy, and corroborated with hematological data. A 5-day and a 30-day treatment with daily doses of 700 µg BV/kg and an acute-lethal treatment with a single dose of 62 mg BV/kg were performed. The 5-day treatment resulted in a reduced cellularity of the bone marrow, with necrosed proerythroblasts, polymorphous erythroblasts, and reticulocytes with cytoplasmic extensions, and a lower number of larger RBCs, with poikilocytosis (acanthocytosis) and anisocytosis, and reduced concentrations of hemoglobin. After the 30-day treatment, the bone marrow architecture was restored, but polymorphous erythroblasts and reticulocytes with thin extensions could still be observed, while the RBCs in higher number were smaller, many with abnormal shapes, especially acanthocytes. The acute treatment produced a partial depopulation of the bone marrow and ultrastructural changes of erythroblasts including abnormal mitochondrial cristae. The RBCs in lower number were bigger and crenated, with reduced concentrations of hemoglobin. Overall, BV was able to promote stress erythropoiesis in a time- and dose-related manner, mitochondrial cristae modification being a critical factor involved in the toxicity of the BV high doses.

Identification of new structural elements within 'porosomes' of the exocrine pancreas: a detailed study using high-resolution electron microscopy.

In the past two decades, great progress has been made in our knowledge of how cells secrete. This progress has been possible primarily due to discovery of the 'porosome', the universal secretory portals at the plasma membrane in cells. Porosomes are permanent cup-shaped lipoprotein structures at the cell plasma membrane, where membrane-bounded secretory vesicles temporarily dock and fuse to expel all or part of their contents during cellular secretion. Porosomes have been found in neurons, in neuroendocrine cells, as well as in the exocrine pancreas. Furthermore, porosomes have been isolated, functionally reconstituted, and their composition determined. Although, the neuronal porosome has been exhaustively investigated, the detailed morphology of porosomes in the exocrine pancreas in situ remains to be further explored. The current study was carried out to determine the detailed morphology of the porosome in rat exocrine pancreas using high-resolution electron microscopy. Results from our study, demonstrate for the first time the presence of tethers or cables (which could be t-SNAREs) associated at the base of porosomes. Furthermore, for the first time our studies demonstrate the docking of a single secretory vesicle at the base of more than one porosome complex. Detailed spoke-like elements lining the porosome cup are also demonstrated for the first time in our study, providing a better understanding of the molecular architecture and physiology of this important cellular organelle.

Bee (Apis mellifera) venom produced toxic effects of higher amplitude in rat thoracic aorta than in skeletal muscle--an ultrastructural study.

In this study, changes produced in aorta and triceps surae muscle of Wistar rats as response to bee venom (BV) envenomation were analyzed by transmission electron microscopy and morphometry. A subchronic treatment of 30 days with daily doses of 700 µg BV/kg and an acute-lethal treatment with a single dose of 62 mg BV/kg were performed. The subchronic treatment resulted in endothelial cell retraction, a thicker subendothelial layer, and thinner elastic laminae and musculoelastic layers in aorta, and thicker endothelium and basal laminae in skeletal muscle. In both tissues polymorphous, swollen mitochondria with disrupted cristae were observed. The acute treatment produced extensive endothelial lesions, breakdown of the collagen layer and migration of muscle cells toward the intima in the aorta, and dilatation of endoplasmic reticulum in the skeletal muscle cells. Mitochondria were almost devoid of cristae or with few circular cristae in the smooth muscle cells while most of the mitochondria presented abnormal circular cristae in the skeletal muscle cells. Degenerative alterations in the aorta were of higher intensity in our experiments-both the intima and media strongly responded to BV, in contrast to those found at the level of the skeletal muscle cells where a moderate degenerative myopathy was recorded.

Electrophysiological and structural aspects in the frontal cortex after the bee (Apis mellifera) venom experimental treatment.

The aim of this study is to evaluate the bioelectrical and structural-functional changes in frontal cortex after the bee venom (BV) experimental treatments simulating both an acute envenomation and a subchronic BV therapy. Wistar rats were subcutaneously injected once with three different BV doses: 700 µg/kg (T(1) group), 2100 µg/kg (T(3) group), and 62 mg/kg (sublethal dose-in T(SL) group), and repeated for 30 days with the lowest dose (700 µg/kg-in T(S) group). BV effects were assessed by electrophysiological, histological, histochemical, and ultrastructural methods. Single BV doses produced discharges of negative and biphasic sharp waves, and epileptiform spike-wave complexes. The increasing frequency of these elements suggested a dose-dependent neuronal hyperexcitation or irritation. As compared to the lower doses, the sublethal dose was responsible for a pronounced toxic effect, confirmed by ultrastructural data in both neurons and glial cells that underwent extensive, irreversible changes, triggering the cellular death. Subchronic BV treatment in T(S) group resulted in a slower frequency and increased amplitude of cortical activity suggesting neuronal loss. However, neurons were still stimulated by the last BV dose. Structural-functional data showed a reduced cellular density in frontal cortex of animals in this group, while the remaining neurons displayed both specific (stimulation of neuronal activity) and unspecific modifications (moderate alterations to necrotic phenomena). Molecular mechanisms involved in BV interactions with the nervous tissue are also discussed. We consider all these data very important for clinicians who manage patients with multiple bee stings, or who intend to set an appropriate BV therapy.

Abnormal mitochondrial cristae were experimentally generated by high doses of Apis mellifera venom in the rat adrenal cortex.

In the present study, Apis mellifera venom (AmV) was tested for its ability to cause ultrastructural changes in mitochondria of rat adrenal cortex in vivo. In order to achieve this goal, different AmV treatments were performed and the effects were quantified on transmission electron micrographs. In a first experimental group, AmV injected for 30 days in low daily doses (700 µg/kg) generated important ultrastructural changes in zona fasciculata. The mitochondrial ultrastructure was not affected, but the diameters of mitochondrial cristae (MC) were reduced (57.066 ± 7.795 nm) as compared to the MC diameters in the corresponding control groups (58.596 ± 6.603 nm, and 58.503 ± 5.708 nm). In adrenal glands collected from rats injected with a single, high dose of AmV (62 mg/kg), many ultrastructural changes were described. Mitochondria with normal, tubular MC (with diameter of 58.711 ± 5.907 nm) were observed in many cells, very close to the values calculated for the corresponding control group (58.639 ± 6.117 nm). However, the striking data reported herein concerned the ability of AmV high doses to promote dramatic alterations in the ultrastructure of these particular mitochondria, similar to those described in certain severe diseases. Thus, several types of abnormal mitochondria were observed, including mitochondria displaying lamellar and/or circular, concentric cristae and mitochondria devoid of cristae. The abnormal circular, concentric MC, with large inner (281.904 ± 158.588 nm) and outer (432.076 ± 230.372 nm) diameters, appeared to be the most stable form of MC in the adrenal cortex after the acute treatment with AmV. Among other ultrastructural aspects, these important changes indicated a high level of cytotoxicity of AmV in adrenocortical cells following in vivo experimental poisoning.

Ultrastructural evidence of mucosal healing after infliximab in patients with ulcerative colitis.

BACKGROUND: Infliximab is a monoclonal anti-TNF-alpha antibody that has been shown to be effective in Crohn's disease therapy. However, data are scarce about the mechanism of action and its efficacy in ulcerative colitis (UC). AIM: To assess intracellular changes of the colonic mucosa in patients with UC before and after infliximab treatment. METHODS: 7 patients (18-65 years, 4 men) with active, refractory, moderate to severe UC (Lichtiger's Clinical Activity Index > 6, Endoscopic Index > 4) underwent colonoscopy before and 4 weeks after the initial infusion of infliximab 5mg/kg of body weight. Endoscopically obtained biopsy specimens were processed specifically, stained with uranyl-acetate and lead citrate and examined with a JEOL-1010 transmission electron microscope. RESULTS: Before treatment we noticed severe alterations of the epithelium: microvilli depletion, shattering of the epithelial junctions, cytoplasmic vacuolization, dilatation of the endoplasmic reticulum, pycnotic nuclei, altered structure of mitochondria and Golgi complexes. Rarefaction of the goblet cells, and abnormal mucus formation and secretion were also observed. The corresponding chorion showed structural alteration of component cells, obstructed capillaries, erythrocyte extravasation, and many plasmocytes and neutrophils. After infliximab, improvement in morphology and function of the epithelial organelles, rich mucus secretion and recovery of the chorionic components were noticed. CONCLUSIONS: Our study revealed important intracellular alterations of the UC mucosa that were restored after infliximab therapy. These features may contribute to a better understanding of UC pathogenesis and mechanism of action of the anti-TNF-alpha therapies.

Premature termination of hair follicle morphogenesis and accelerated hair follicle cycling in Iasi congenital atrichia (fzica) mice points to fuzzy as a key element of hair cycle control.

Inbred laboratory mice have proven to be useful model systems for studying hair biology and pathomechanisms of hair loss. Fuzzy (fz) is an autosomal recessive mutation that results in hair coat abnormalities. Though this mutant has long been known, its cutaneous abnormalities still await systematic analysis. Here, we provide a systematic skin phenotype analysis of mice that are homozygous for Iasi congenital atrichia (fzica/fzica), which is allelic to fz. Homozygous mice exhibit a sparse hair coat after birth and completely loose their hair at around postnatal day 120. Although early and mid stages of hair follicle morphogenesis are normal, late hair follicle morphogenesis reveals multifocal cell degeneration within the Huxley layer of the inner root sheath (IRS) and a complete lack of the hair shaft medulla. In addition, hair follicle development is prematurely terminated by induction of the first postnatal hair cycle with premature entry into catagen. Subsequently, a dramatically shortened telogen is immediately followed by premature anagen development, resulting in a marked, generalized acceleration of hair follicle cycling. This suggests that fuzzy is not only involved in structural hair shaft integrity and differentiation of the IRS and medulla, but also plays an important role in the control of hair follicle cycling. Our data show that fuzzy is involved in controlling both catagen and anagen initiation, designating fuzzy an exciting target for characterizing the intracutaneous oscillator system that drives hair follicle cycling.

Elucidation of cell secretion: pancreas led the way.

Secretion is a basic process in all cells and is required for several important functions such as neurotransmission, the secretion of digestive juices from the exocrine pancreas and the release of hormones from endocrine and neuroendocrine cells. Due to these important functions, the mechanism of cell secretion has been intensely investigated for over half a century. However, it is only in the last decade, with the discovery of a new cellular structure, the 'porosome' or 'fusion pore', and the elucidation of SNARE-induced membrane fusion, that has finally provided us with an understanding of cell secretion at the molecular level. The 'porosome', a supramolecular structure at the cell plasma membrane, was first discovered in the exocrine pancreas, and subsequently in endocrine/neuroendocrine cells and in neurons. The structure and dynamics of the 'porosome' in live cells at nanometer resolution and in real-time, its composition and functional reconstitution in lipid membrane, have all been determined. These findings have fundamentally changed our understanding of cell secretion and provide a clear understanding of this highly regulated process in cells.