Cisternae Of Granular Endoplasmic Reticulum Research Articles

Functional ultrastructure and cytochemistry of vitellogenesis stages of Rohdella amazonica (Aspidogastrea, Aspidogastridae, Rohdellinae), a parasite of the Amazoninan banded puffer fish Colomesus psittacus

Vitellogenesis of Rohdella amazonica, parasites of the intestinal tract of a naturally infected banded puffer fish Colomesus psittacus, were studied by transmission electron microscopy (TEM) and TEM cytochemistry. Four vitellogenic stages were distinguished (I) stem cell stage of the gonial type (II) early differentiation stage, characterized by initiation of protein synthetic activity (III) an advanced differentiation and vitellocyte maturation stage, involving rapid accumulation of β-glycogen, large lipid droplets and smaller shell globules that fused progressively into large electron-dense shell globule clusters (IV) mature vitellocyte stage. The vitellogenesis process involves substantial increase in cell volume; extensive development of parallel cisternae of granular endoplasmic reticulum (GER) that become concentrically arranged in later stages and secrete shell globules; development of Golgi complexes engaged in the packaging of shell material; progressive accumulation, fusion and enlargement of unsaturated and saturated lipid droplets; formation of β-glycogen and α-glycogen and their accumulation into islands scattered among lipid droplets in maturing and mature vitellocytes; and finally during the advanced stage of maturation, rapid accumulation of large saturated lipid droplets α- and β-glycogen, and proteinaceous shell-globules and clusters in the peripheral cytoplasmic layer. Vitellogenesis in R. amazonica generally resembles that previously described for bothriocephalidean cestodes, but differs from that of other cestode orders. Positive cytochemical staining with periodic acid-thiocarbohydrazide-silver proteinate confirms β-glycogen and α-glycogen, around saturated lipid droplets of maturing and mature vitellocytes. These results from a basal trematode lead to a better understanding of the phylogenetic, functional, and developmental relationships among vitellogenesis patterns as they relate to egg formation, embryonic development, and life cycles among trematodes and cestodes.

The effects of the anti-Müllerian hormone on folliculogenesis in rats: light and electron microscopic evaluation

ABSTRACT In this study, we evaluated the effects of anti-Müllerian hormone on follicle development and oocyte quality with light and electron microscopy. Twenty-four adult female rats were divided into four groups. After estrous cycle synchronization, on the first day, control group rats were injected with 0.5 ml saline, 2nd, 3rd, and 4th groups were injected 1 µgr, 2 µgr, and 5 µgr anti-Müllerian hormone, respectively. On the third day, intracardiac blood samples were taken for follicle-stimulating hormone, luteinizing hormone, estradiol, and progesterone serum level measurements. Ovaries were obtained for light and electron microscopic examinations. Secondary (antral) follicles were decreased while atretic follicles were increased in number parallel with an increased dose of anti-Müllerian hormone injection. Atresia of the follicles was demonstrated with apoptosis of granulosa cells characterized by apoptotic bodies and with paraptosis characterized by the vacuole formation in the cytoplasm, enlargement of granular endoplasmic reticulum cisternae and perinuclear cisternae in granulosa cells. Premature luteinization characterized by increased lipid droplets, mitochondria with tubular cristae, and smooth-surfaced endoplasmic reticulum in the cytoplasm of granulosa cells were detected in some growing follicles. In the anti-Müllerian hormone injected experimental groups, cystic follicles characterized by a large antrum, attenuated granulosa cell layer, and flattened granulosa cells that face the antrum were observed. Corpus luteum and stroma were similar in all groups. It was concluded that increasing doses of anti-Müllerian hormone caused increased atresia in developing follicles, premature luteinization of granulosa cells in some follicles, and cystic follicle formation in the further developing follicles.

THE CHANGES OF WOUND MACROPHAGES IN PATIENTS WITH DIABETES

Purulent-necrotic lesions of the extremities require amputation in 30-50% of cases. Among all cases of lower extremity amputations 50-70% are due to diabetes. Moreover, 5 out of 6 amputations, not related to the traumatic injury of the limb, are performed in patients with diabetes. The mortality rate among patients with diabetes, who undergo amputation varies from 28 to 40%, and 5-year surveillance is only 10-25%. The study of ultrastructural changes of macrophages on the 3rd day of treatment revealed masses of chaotically located fibrillar structures in the cytoplasm of macrophages that occasionally had an increased electron density. This phenomenon was observed in the purulent-necrotic areas of soft tissues of patients from the main group, compared to the control group. In all cases, mitochondria were enlarged in size, swollen, with a light matrix and contained a reduced amount of cristae. The cristae were deformed and shortened. Swollen matrix in mitochondria led to the formation of vacuoles on their place containing fine-grained contents. The nucleus had a usual form and size with the presence of single invaginations. Chromatin was predominantly concentrated in the form of solid electron-dense masses or evenly distributed throughout the nucleus. There were nuclei with partial chromatin dispersion. The contents of the nuclei included granular, fibrillar, and fine vacuolar material. The nuclear membrane folding did not fluctuate significantly. The folds did not cover the whole surface of the nucleus. In some areas invaginates were represented by the continuation of perinuclear space only. The nuclear envelop pores, which connect the contents of the cytoplasm and nucleoplasm, have been observed. The cytoplasm between the zones of the plate complex was occupied by small mitochondria, single polysomal rosettes, and cisternae of granular endoplasmic reticulum, which was represented by extended intracellular channels and vacuolar formations. The smooth endoplasmic reticulum was predominantly located in the central part. The surface of macrophages in the process of their differentiation from monocytes was relatively plane. Occasionally there occurred small processes or pseudopodia. The number of pinocytic vesicles surrounded by a border was reduced in poorly differentiated cells. Ozone therapy stimulates the functional activity of wound macrophages, as it causes destructive changes in these cells without necrotic lesions. Intravenous introduction of ozonized saline con-tributes to the elimination of wound macrophag-es, mainly through genetically programmed cell death (apoptosis), which plays a significant role in the regulatory mechanisms of the inflammato-ry process.

Ultrastructure of the uterus, embryonic envelopes and the coracidium of the enigmatic tapeworm Tetracampos ciliotheca (Cestoda: Bothriocephalidea) from African sharptooth catfish (Clarias gariepinus).

Transmission electron microscopy (TEM) was used to study the ultrastructure of the uterus and egg morphology in the enigmatic bothriocephalidean tapeworm Tetracampos ciliotheca. The uterine wall, underlain by well-developed muscle bundles, consists of a syncytial epithelium which is characterized by the abundance of free ribosomes, mitochondria and cisternae of granular endoplasmic reticulum (GER). On the apical surface of the uterine epithelium, there is an abundant network of cytoplasmic microlamellae projecting into the uterine lumen. The lumen is filled with freely lying eggs which are located close to the uterine wall but do not contact with the microlamellae of the uterine epithelium. The developed eggs possess an oncosphere surrounded by four envelopes: (1) a thin egg shell; (2) an outer envelope; (3) a syncytial, ciliated inner envelope; and (4) the oncospheral membrane. The mature hexacanth is armed with three pairs of oncospheral hooks, as well as somatic and hook muscles and five types of cells (1) binucleated subtegumental cell, (2) somatic cells, (3) penetration gland cells, (4) nerve cells and (5) germinative cells. Considering the relative scarcity of descriptive and comparative studies on the ultrastructure of the uterus and egg morphology in the order Bothriocephalidea, we concluded that Tet. ciliotheca displays a unique type of egg development. Based on these results, we discuss plausible ideas relating to the function of these structures for consideration in future studies.

Serum adipokine concentrations in patients with type 2 diabetes: the relationships with distribution, hypertrophy and vascularization of subcutaneous adipose tissue

Academy of Sciences, Novosibirsk, Russia 2Novosibirsk State Medical University, Novosibirsk, Russia
 BACKGROUND: Adipose tissue (AT) dysfunction plays an important role in metabolic disorders in obesity and type 2 diabetes. The role of distribution, hypertrophy and vascularization of AT in adipokine secretion disturbances remain to be clarified.
 AIMS: To determine the relationships between serum concentrations of adipokines and the mass and distribution of AT, diameter of adipocytes and vascularization of subcutaneous AT in patients with type 2 diabetes.
 MATERIALS AND METHODS: A total of 125 patients were examined, including 82 subjects with obesity. Thirty persons without diabetes and obesity, matched by sex and age, were acted as control. Concentrations of leptin, resistin, visfatin, adipsin and adiponectin in fasting serum were determined using multiplex analysis. Mass and distribution of AT was assessed by dual-energy X-ray absorptiometry. Samples of SAT were obtained from umbilical region using a knife biopsy in 25 patients and in 15 individuals who died in accidents. Blood and lymphatic vessels in SAT were revealed with immunohistochemistry, using antibody to CD-34 and podoplanin respectively. The volume and numerical density, ultrastructure of blood and lymphatic vessels, and mean diameter of subcutaneous adipocytes were evaluated.
 RESULTS: Patients with diabetes, as compared to control, had significantly higher levels of leptin, resistin, adipsin and visfatin (all p0.001). Adiponectin showed no differences. Concentrations of leptin, resistin, visfatin, adipsin and adiponectin correlated positively with gynoid fat mass. Additionally, leptin and adipsinshowed positive correlations with truncal and central abdominal fat mass. Concentration of leptin, but not other adipokines, was associated with hypertrophy of subcutaneous adipocytes. A decrease in volumetric density of microvessels(р=0.01) and increase in volume and numerical density of lymphatic vessels (both р=0.02) was observed in subcutaneous AT from diabetic subjects. The swelling of cytoplasm, mitochondria, cisterns of granular endoplasmic reticulum and reduced content of micropinocytotic vesicles was revealed in lymphatic capillaries. Resistin and visfatin showed inverse associations with density of microvessels.
 CONCLUSION: Endocrine dysfunction of AT in patients with type 2 diabetes, manifested by elevation of serum concentrations of leptin, resistin, visfatin and adipsin, is associated with mass and distribution of AT, hypertrophy of subcutaneous adipocytes and vascularization abnormalities of subcutaneous AT.

Dysregulation of placental ABC transporters in a murine model of malaria-induced preterm labor

Malaria in Pregnancy (MiP) is characterized by placental accumulation of Plasmodium-infected erythrocytes, intrauterine growth restriction (IUGR) and preterm delivery (PTD). Placental ATP-binding cassette (ABC) transporters mediate the efflux of nutrients, cytokines and xenobiotics. The expression and activity of these transporters are highly responsive to infection. We hypothesized that MiP would perturb the expression of placental ABC transporters, promoting PTD. Peripheral blood, spleens, livers and placentas of pregnant mice, infected with Plasmodium berghei ANKA on gestational day (GD) 13.5, were collected and analyzed on GD18.5. The primary consequences of human MiP, including IUGR, PTD (20%) and placental inflammation, were recapitulated in our mouse model. Electron microscopy revealed attenuated presence of labyrinthine microvilli and dilated spongiotrophoblasts -granular endoplasmic reticulum cisternae. Additionally, a decrease in placental Abca1 (ABCA1), Abcb1b (P-glycoprotein), Abcb9 and Abcg2 (BCRP) expression was observed in MiP mice. In conclusion, MiP associated with PTD impairs placental ABC transporters’ expression, potentially modulating placental nutrient, environmental toxin and xenobiotic biodistribution within the fetal compartment, and may, at some degree, be involved with pregnancy outcome in MiP.

Fine structure of the uterus in tapeworm Tetrabothrius erostris (Cestoda: Tetrabothriidea).

The uterine organization in Tetrabothrius erostris (Tetrabothriidea) was investigated by the methods of transmission and scanning electron microscopy. In sexually mature proglottids, the uterine wall consists of a syncytial epithelium (1.4-2.5 μm thick, except in regions containing nuclei). The ribosomes, mitochondria and numerous cisternae of granular endoplasmic reticulum with concentric or parallel profiles with electron lucent material are observed in the epithelium. The uterine wall is characterized by the abundance of lipid droplets that are localized inside the long protrusions of the uterine epithelium (called fungiform papillae) up to 15-17μm and in the surrounding medullary parenchyma. The protrusions with lipid droplets in the proximal ends of the uterus are located closely to each other. A basal matrix (up to 0.6 μm thick) supports the uterine epithelium. The musculature consisting of 1-2 muscle layers is well developed; large myocytons are connected with the myofibrils and have a nucleus that reaches 4μm in size. In gravid proglottids, the epithelium without nuclei is reduced to 0.2-1.6 μm thick. The number of protrusions of the uterine epithelium and lipid droplets in the epithelial layer decreases. Sparse small muscle bundles underlay the uterine wall at this stage; the basal matrix is feebly marked. The matrotrophy or the support by nutrition from the parent organism to embryos is discussed for T. erostris which belongs to oligolecital cestodes and possesses numerous lipid droplets in the uterine wall during the development of embryos.

Vitellogenesis in Orientocreadium batrachoides Tubangui, 1931 (Digenea: Allocreadiidae) from Clarias gariepinus, an ultrastructural study

Vitellogenesis in Orientocreadium batrachoides, an intestinal parasite of the catfish, Clarias gariepinus was investigated by transmission electron microscope. Four stages of vitellocyte development have been distinguished during vitellogenesis: (I) stem cell stage of the gonial type (immature vitellocyte), (II) early and (III) advanced stages of maturing vitellocytes. Both early and advanced stages of maturing vitellocytes involved the initiation of protein synthetic activity, onset of shell globule formation and progressive fusion of single shell globules into large shell globule cluster in their cytoplasm and (IV) mature vitellocyte. Vitellocyte maturation is characterized by: (1) an increase in cell volume, (2) extensive development of parallel cisternae of granular endoplasmic reticulum, (3) development of Golgi complexes and (4) a continuous development of shell globules that fused into shell globule clusters. Mature vitelline cells are characterized by shell globule clusters, which play an important role in egg shell formation, lipid droplets and β-glycogen that accumulated in large amounts in their cytoplasm. Single “lamellar” granules are present in the cytoplasm of the mature vitellocytes.

Light and electron microscopic study on vitellogenesis of the caryophyllidean cestode Monobothrioides chalmersius (Woodland, 1924) Hunter, 1930 (Lytocestidae) from the catfish Clarias gariepinus

Transmission electron microscopy (TEM) has been applied for the first time to study vitellogenesis in the caryophyllidean tapeworm Monobothrioides chalmersius (Woodland, 1924) Hunter, 1930 from the catfish Clarias gariepinus inhabiting Nile Delta water in Egypt. During development of vitellocytes, cell size and nuclear surface are increased. Meanwhile, parallel cisternae of granular endoplasmic reticulum (GER) and Golgi complexes develop, but the nucleo-cytoplasmic ratio is restored, shell-globules (vitelline granules) are formed, small shell-granules fuse into larger shell-globules and large shell-globules fuse into larger shell-globule clusters, cytoplasmic and nuclear glycogen accumulate. Mature vitellocytes of M. chalmersius contain five kinds of vitelline material: shell globules clusters (vitelline granules), glycogen, few probably lipid droplets, many peripheral translucent vesicles and electron-dense vesicles. The interstitial tissue consists of cells from which many cytoplasmic processes extend to be in close contact with maturing and mature vitellocytes. The cytoplasm of the interstitial cells contains some electron-dense vesicles. The possible function of the interstitial cells inclusions is discussed. The vitellogenesis of M. chalmersius follows the basic pattern of the process in caryophyllidean cestodes but some differences were recorded particularly in the ultrastructure of the mature vitellocytes and interstitial tissue. Vitellogenesis and ultrastructure of the mature vitellocytes and interstitial tissue of M. chalmersius are compared particularly with those in other monozoic cestodes.

Crohn’s disease of the colon: ultrastructural changes in submuscular interstitial cells of Cajal

Interstitial cells of Cajal (ICC) at the submuscular border of the human colon (ICC-SMP) are the proposed pacemaker cells of the musculature. In patients with Crohn's disease (CD) of the colon, ICC-SMP showed characteristic cytological changes from controls. The changes comprised secondary lysosomes in connection with lipid droplets and cytoplasmic vacuoles or multiple empty, confluent and often outbulging vacuoles merging with cisterns of granular endoplasmic reticulum and clusters of glycogen granules. These changes were most pronounced in patients with macroscopical mucosal inflammation but were also demonstrable in uninvolved colonic segments. Relationships of ICC to other cells were undisturbed. The changes were selective to ICC-SMP, as glial cells, muscle cells and fibroblast-like cells at the submuscular border showed no cytological alterations compared with controls. Varicosities of the submuscular plexus were often empty and dilated. Fibroblast-like cells selectively encased macrophages and mast cells. The cytological changes in ICC-SMP in CD are thus similar to changes seen in ulcerative colitis and may be of pathophysiological significance with regard to the motility and sensory disturbances seen in patients with CD.

Electron microscopical study to assess thein vitroeffects of the synthetic trioxolane OZ78 against the liver fluke,Fasciola hepatica

Adult Fasciola hepatica were incubated for 48 h in vitro in the synthetic peroxide, OZ78 at a concentration of 100 microg/ml and then prepared for scanning and transmission electron microscopy. There was limited disruption to the external fluke surface, with only slight swelling and blebbing of the interspinal tegument in the midbody and ventral tail regions. By contrast, significant disruption was observed to the ultrastructure of the tegument and subtegumental tissues. There was severe swelling of the basal infolds in the tegumental syncytium and the flooding spread internally to affect the subtegumental tissues. In the tegumental system, there was swelling of the cisternae of granular endoplasmic reticulum and of the mitochondria, with the latter showing signs of breaking down. Autophagic vacuoles and lipid droplets were present and the synthesis of tegumental secretory bodies was much reduced. The gastrodermal cells were severely affected, with swelling and degeneration of the mitochondria and the presence of autophagic vacuoles and lipid droplets. The granular endoplasmic reticulum was swollen and vesiculated and the cells contained few secretory bodies. Both the vitelline and testis follicles showed evidence of extensive cellular disruption and degeneration. This study confirms previous data indicating the potential flukicidal activity of OZ78.

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.

Ultrastructural changes in rat thyroid tissue after acute organophosphate poisoning and effects of antidotal therapy with atropine and pralidoxime: A single-blind, ex vivo study

Background: Organophosphate (OP) insecticides are widely used in both agricultural and landscape pest control, and the potential for human exposure to these compounds is significant. Objectives: The aims of this study were to investigate the effects of acute poisoning with the OP methamidophos and the effects of antidotal therapy with atropine and pralidoxime on rat thyroid tissue ultrastructure. Methods: In this single-blind, ex vivo study, male Wistar albino rats weighing 220 to 230 g were divided into 4 treatment groups. Group 1 received a median lethal dose of methamidophos (30 mg/kg) via oral gavage. Group 2 received saline via oral gavage and served as the control group for group 1. Group 3 received methamidophos (30 mg/kg) via oral gavage, and after 8 minutes atropine 0.05 mg/kg and pralidoxime chloride (2-FAM) (40 mg/kg) were administered intraperitoneally (IP). Atropine was titrated to reverse signs of cholinergic excess. Group 4 received saline via oral gavage followed by IP injections and served as the control for group 3. Rat thyroid tissues were examined using electron microscopy, and the histologic changes were examined by a histopathologist who was blinded to treatment. All rats were euthanized by intracardiac blood collection. The rats in groups 1 and 2 were euthanized 8 minutes after treatment. The rats in groups 3 and 4 were euthanized 96 hours after treatment. Results: Thirty-four male rats (aged 16 weeks) were included in the study. The rats were grouped accordingly: group 1 (n = 10); group 2 (n = 7); group 3 (n = 10); and group 4 (n = 7). The mean (SD) pseudocholinesterase (FCE) activity was significantly lower in the methamidophos-treated rats (group 1) compared with the corresponding control group (group 2) (32.6 [17.0] vs 579.4 [59.0] U/L, respectively; P < 0.001). PCE activity was significantly higher in rats treated with atropine and 2-PAM (group 3) (392.5 [39.4] U/L; P < 0.001) compared with those not receiving antidotal therapy (group 1). Group 1 experienced changes in thyrocytes and organelles that were not detected in the antidote-treated rats in group 3. These changes included follicular cell nuclei exhibiting an increase in chromatin content, pyknotic nuclei, mitochondrial degeneration, dilated granular endoplasmic reticulum cisternae, reduced microvilli, and intraluminal cellular debris. Within follicular cells, formation of vacuoles filled with fine granular material was noted. Conclusion: Acute OP poisoning was associated with histopathologic effects in rat thyroid tissue that appeared to be mitigated by antidotal therapy in this small animal study. More extensive studies using immunohistochemical methods are needed.

Ultrastructural changes induced in the tegument and gut of Fasciola hepatica following in vivo and in vitro drug treatment with nitroxynil (Trodax)

Male Sprague-Dawley rats were dosed orally with nitroxynil at a concentration of 40 mg/kg, and adult Fasciola hepatica were recovered after 24, 48 and 72 h. Fine structural changes to the tegument and gut were monitored by transmission electron microscopy. Flukes were also incubated for 24 h in vitro in nitroxynil at a concentration of 100 microg/ml. Following treatment in vivo, there was an accumulation and accelerated release of secretory bodies at the apex of the tegumental syncytium. Some swelling of the mucopolysaccharide masses surrounding the basal infolds was evident after 48 and 72 h. There was an initial accumulation of T1 secretory bodies at the base of the syncytium, but this decreased at 72 h, coinciding with a decline in their production in the tegumental cells. The mitochondria were consistently swollen in the tegumental cells. At 72 h, large vacuolations were observed between the muscle layers and there was flooding around the underlying tissues. Some tegumental cells were seen to be degenerating and beginning to disintegrate. After 24 h treatment in vitro, the basal infolds were swollen and the crystalline structure of the spines was disrupted. Flooding of the internal tissues was evident and, in the tegumental cells, Golgi complexes and secretory bodies were absent. The mitochondria in the tegumental cells were swollen. In the gastrodermal cells, changes were evident at the earliest time period in vivo. The lamellae were disrupted, few secretory bodies were present, the mitochondria and cisternae of granular endoplasmic reticulum (ger) were swollen and there was an increased number of secretory bodies. These changes became progressively more severe with time. Similar changes were evident following treatment in vitro; vesiculation of the ger was also seen. The results indicate that oral uptake is the predominant route of entry of nitroxynil into the fluke.

Adult triclabendazole-resistant Fasciola hepatica: surface and subsurface tegumental responses to in vitro treatment with the sulphoxide metabolite of the experimental fasciolicide compound alpha

Mature Fasciola hepatica of the triclabendazole-resistant Sligo isolate were incubated in vitro with 10 microg/ml of the sulphoxide metabolite of compound alpha [5-chloro-2-methylthio-6-(1-naphthyloxy)-H-benzimidazole]; the metabolite will be referred to as alpha.SO. Changes resulting from drug treatment were examined by scanning electron microscopy (SEM), transmission electron microscopy (TEM) and tubulin immunocytochemistry (ICC). SEM revealed that disruption to the tegumental surface mainly took the form of swelling and blebbing. Extensive spine loss occurred on the ventral surface of the oral cone, and sloughing of the tegument was observed along the lateral margins of the fluke. Examination of sections from the anterior mid-body region at the TEM level revealed that treatment with alpha.SO led to swelling of the basal infolds and mitochondria within the tegumental syncytium; also, accumulations of secretory bodies beneath the apical plasma membrane. The tegumental cell bodies contained swollen mitochondria and cisternae of granular endoplasmic reticulum, but few Golgi complexes were observed. An increase in T2 secretory bodies was observed, whilst in the T1 tegumental cells, the T1 secretory bodies had decreased in number. Immunocytochemical (ICC) studies showed that incubation with alpha.SO, ABZ.SO and TCBZ.SO did not cause significant changes to the distribution of tubulin within the tegumental syncytium of the Sligo isolate. In contrast, alpha.SO, ABZ.SO and TCBZ.SO caused severe disruption to tubulin organization within the syncytial layer of the TCBZ-susceptible Cullompton isolate. The EM results confirm that compound alpha is a fasciolicide capable of disrupting the tegument of mature TCBZ-resistant F. hepatica; however, this was not accompanied by any change in tubulin immunoreactivity.

Submicroscopic structure of canine articular cartilage

Canine articular cartilage was studied in male dogs at age 1, 4, 5 and 8 years. Samples collected from four hip joints and two humeral joints in each age category were processed by standard methods to be examined by scanning and transmission electron microscopy. The cartilage of both joints was similar in structure. In the superficial cartilage layer of one-year-old animals, individual spindle-shaped chondrocytes in the extracellular matrix were, together with associated collagen fibrils, located parallel to the surface. When viewed by scanning electron microscopy, they were distinctly prominent above the surrounding surface. Changes in the thickness and arrangement of both the chondrosynovial membrane and intercellular matrix were apparent in the 4-, 5- and 8-year-old animals, indicating the onset or progression of an osteoarthritic process. The middle cartilage layer in young animals showed elliptical chondrocytes occurring in pairs. The voluminous cytoplasm contained a great amount of granular endoplasmic reticulum, a large Golgi complex and numerous transport vesicles. The pericellular matrix, up to 1 &amp;micro;m thick, was composed of aperiodic fibrils. In the old animals the pericellular matrix was absent and was replaced by thick collagen fibrils with a marked periodicity. The deep cartilage layer in young dogs included groups of three to four chondrocytes situated in a common territory. The cytoplasm contained distinct bundles of intermediary filaments. The pericellular matrix occasionally formed septa between adjoining cells. The intracellular matrix included bundles of collagen fibrils arranged in a matted structure. In the old animals aggregation of chondrocytes into groups almost disappeared. The cytoplasm showed only short cisternae of granular endoplasmic reticulum, small numbers of mitochondria and transport vesicles, frequent lipid droplets and small glycogen deposits. The intercellular matrix consisted of only short collagen fibrils with no distinct periodicity.

Transmission electron microscope study of the ultrastructural changes induced in the tegument and gut of Fasciola hepatica following in vivo drug treatment with clorsulon.

Using transmission electron microscopy (TEM), both the tegument and gut of Fasciola hepatica were examined in an effort to identify and characterise the ultrastructural changes induced following treatment with the flukicidal drug clorsulon. Male Sprague-Dawley rats infected with F. hepatica were dosed orally at 8-8.5 weeks post-infection with clorsulon at a concentration of 12.5 mg/kg body weight. After 24, 48 and 72 h, rats were sacrificed by cervical dislocation and mature flukes recovered from the bile ducts. After 24 h treatment in vivo, disruption of the tegumental syncytium was concentrated at the apex of the syncytium where a dark band consisting of numerous secretory bodies was present. Some blebbing of the apex had also occurred, "open" bodies were present in this region and the mitochondria were slightly swollen. In the cell bodies, swelling of the mitochondria and their cristae had also occurred and the Golgi complexes appeared to be smaller than normal. The disruption seen after 48 h treatment in vivo was similar but more severe: the frequency of blebbing had increased, as had the number of "open" bodies and the swelling of the mitochondria. Vacuoles had begun to appear in the syncytium-both autophagic and electron-lucent-and swelling of the mucopolysaccharide masses around the basal infolds had occurred. Lipid droplets were observed occasionally. In the cell bodies, autophagic vacuoles had begun to appear and swelling of the mitochondria had increased in severity. After 72 h treatment in vivo, more severe disruption was seen in the tegumental syncytium in which widespread swelling and blebbing of the apex was apparent. The basal infolds had become very badly swollen in a number of specimens and damage to the spines was evident. The mitochondria remained swollen, as did the mucopolysaccharide masses around the basal infolds. Lipid droplets were more frequently observed in the syncytium. In the tegumental cells, swelling of the mitochondria was greater and an increase in the number of autophagic vacuoles was apparent. The gut showed signs of disruption after 24 h treatment in vivo, in that the surface lamellae were disrupted and a build-up of autophagic vacuoles at the apex of the cells had taken place. Swelling of the mitochondria and the cisternae of granular endoplasmic reticulum (gER) was evident. There was a decrease in the number of secretory bodies. After 48 h treatment in vivo, the number of autophagic vacuoles in the gastrodermal cells had increased, the mitochondria and gER remained swollen and the disruption seen to the lamellae was still evident. In the 72 h-treated specimens, the disruption seen in the gastrodermal cells had increased significantly, with severe vacuolation of the apical cytoplasm. An increase in the number of autophagic vacuoles was evident, the mitochondria and the gER remained swollen and lipid droplets were present in the cells.

Protective effects of ascorbic acid, dl-alpha-tocopherol acetate, and sodium selenate on ethanol-induced liver damage of rats.

In this study, the effect of a combination of vitamin C (ascorbic acid), vitamin E (dl-alpha-tocopherol acetate), and selenium (sodium selenate) on ethanol-induced liver damage in rats was investigated, morphologically and biochemically. The ethanol-induced injury was produced by the administration of 1 mL of absolute ethanol to each rat. Animals received vitamin C (250 mg/kg), vitamin E (250 mg/kg), and selenium (0.5 mg/kg) (ViCESe) for 3 d 1 h prior to the administration of absolute ethanol. In the liver of the animals given ethanol, the degenerative changes such as extreme hyperemia, vacuolization in cells of portal areas, a dilation in sinusoids, mononuclear cell infiltration, a swelling in cisternae of granular endoplasmic reticulum and in mitochondrial cristae, an increase in smooth endoplasmic reticulum, many lipid vacuoles were observed both light and electron microscopically. A similar structure was usually distinguished when compared with control animals, in rats given ethanol + ViCESe. In this group, the findings indicating cellular damage were either not observed at all or were decreased. In the group administered ethanol, a reduction of the blood glutathione (GSH) level and increases in serum values of alanine aminotranserase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH), alkaline phosphatase (ALP), and gamma-glutamyl transferase (GGT) activities were observed, whereas in the control group, the reverse was found to occur. On the other hand, in the group in which ethanol + ViCESe was administered, it was observed that the blood GSH value and serum ALP and ALT activities increased and serum AST, LDH, and GGT activities decreased. As a result, the present study indicates that ViCESe because of their antioxidant activity against ethanol damage have a protective effect on the liver.

Effects of chard ( Beta vulgaris L. var. cicla) extract on pancreatic B cells in streptozotocin-diabetic rats: a morphological and biochemical study

Chard ( Beta vulgaris L. var. cicla) is used as a hypoglycemic agent by diabetic patients in Turkey. The present study was carried out in order to detect whether this plant, used in folk remedies for decreasing blood glucose levels, affects pancreatic B cells and blood glucose. In the diabetic group, a decrease in the number of B cells of Langerhans islets and in the secretory materials, a swollen granular endoplasmic reticulum cisternae and widened intercellular areas in some of B cells were observed. But, in a diabetic group given chard extract, an increase in the number of B cells of Langerhans islets and in the secretory granules were noted, together with many hypertrophic Golgi apparatus and granules of low densities. The extract while having no effect on blood glucose and body weight in the normal group, reduced the blood glucose value in streptozotocin-induced hyperglycemic animals. But, in a diabetic group given chard, the body weight significantly increased in comparison to the diabetic group; maximum reduction in blood glucose levels was observed on the 42nd day. According to the morphological and biochemical results obtained, it is concluded that the extract of this plant when administered by gavage may reduce blood glucose levels by regeneration of the B cells.

Ultrastructure studies of Proteocephalus longicollis (Cestoda, Proteocephalidea): transmission electron microscopy of scolex glands.

The ultrastructure of two types of secretory glands in the scolex of preadults of Proteocephalus longicollis is described for the first time in the present report. The gland cells contain extensive cisternae of granular endoplasmic reticulum and Golgi complexes, which participate in the production of secretory globules. Type I scolex glands produce electron-dense globules of various size. The secretory globules enter the secretory canal, openings of which were not observed in the preadults. The secretory product of type I was found at the inner sucker surface and in the tegument of the sucker edges. In addition, electron-dense globules in adult worms are secreted via an eccrine mechanism. Type II scolex glands are characterized by secretory globules of lower electron density and occur mainly in preadults. The electron-lucent, membrane-bound secretory globules are transported via microtubule-lined ducts opening to the exterior at the tegumental surface. Secretory globules of type II are released by an eccrine process.