Well-developed Endoplasmic Reticulum Research Articles

Endoplasmic Reticulum Stress in Hepatitis B Virus and Hepatitis C Virus Infection.

Endoplasmic reticulum (ER) stress, a type of cellular stress, always occurs when unfolded or misfolded proteins accumulating in the ER exceed the protein folding capacity. Because of the demand for rapid viral protein synthesis after viral infection, viral infections become a risk factor for ER stress. The hepatocyte is a cell with large and well-developed ER, and hepatitis virus infection is widespread in the population, indicating the interaction between hepatitis viruses and ER stress may have significance for managing liver diseases. In this paper, we review the process that is initiated by the hepatocyte through ER stress against HBV and HCV infection and explain how this information can be helpful in the treatment of HBV/HCV-related diseases.

The Interplay between Finasteride-Induced Androgen Imbalance, Endoplasmic Reticulum Stress, Oxidative Stress, and Liver Disorders in Paternal and Filial Generation.

Finasteride (Fin) causes androgen imbalance by inhibiting the conversion of testosterone (T) to its more active metabolite, dihydrotestosterone (DHT). Androgen receptors (AR) are present (e.g., in hepatocytes), which have well-developed endoplasmic reticulum (ERet). Cellular protein quality control is carried out by ERet in two paths: (i) unfolded protein response (UPR) and/or (ii) endoplasmic reticulum associated degradation (ERAD). ERet under continuous stress can generate changes in the UPR and can direct the cell on the pathway of life or death. It has been demonstrated that genes involved in ERet stress are among the genes controlled by androgens in some tissues. Oxidative stress is also one of the factors affecting the functions of ERet and androgens are one of the regulators of antioxidant enzyme activity. In this paper, we discuss/analyze a possible relationship between androgen imbalance in paternal generation with ERet stress and liver disorders in both paternal and filial generation. In our rat model, hyperglycemia and subsequent higher accumulation of hepatic glycogen were observed in all filial generation obtained from females fertilized by Fin-treated males (F1:Fin). Importantly, genes encoding enzymes involved in glucose and glycogen metabolism have been previously recognized among UPR targets.

Microscopic anatomy of gonadal area in the deep-sea clam Calyptogena pacifica (Bivalvia: Vesicomyidae) with emphasis on somatic cells

Somatic cells in the gonadal area of male and female deep-sea clams, Calyptogena pacifica, were examined using light and transmission electron microscopy. Acini both at the pre-spawning stage and at the stage of active spermatogenesis were observed to be simultaneously present in sections through a male gonad. Oocytes of various degrees of maturity were simultaneously present in female acini. No storage tissue and cells similar to adipogranular cells or vesicular connective tissue of other mollusks were found in the gonadal area of C. pacifica. Instead, in both males and females, numerous hemocytes surround the acini. Among hemocytes, two types of granulocytes and erythrocytes were identified. Bundles of muscle cells were also found in the interacinar space. Male intraacinar accessory cells were rather large, glycogen-rich, with lipid inclusions, and phagosomes contained spermatogenic cells. Female accessory cells had well-developed endoplasmic reticulum, but they did not form any follicles around oocytes, being in their basal part, closer to basal lamina. Such a specific pattern of gonad organization can presumably be explained by both symbiosis with sulfide-oxidizing bacteria and phylogenetic aspects that should be further studied. Some evidence for continuous gametogenesis are discussed.

Ovary Structure and Oogenesis of Trypophloeus klimeschi (Coleoptera: Curculionidae: Scolytinae)

Simple SummaryColeoptera is the largest animal taxon, with many species being agricultural and forest pests. The phylogeny of these species has aroused great interest among scientists. The characteristics of ovariole structure and ultrastructure are useful for phylogenetic work, especially with the improvement of micro technology in recent years. The types of ovarioles are varied. Usually, each family conforms to one type or another. However, in this study, we report on the ovaries of Trypophloeus klimeschi (Coleoptera: Curculionidae: Scolytinae), and find a different ovariole type from that of other known species of Curculionidae. We describe the features of the female reproductive system, ovary and oogenesis of T. klimeschi and also compare these features with those found in other Curculionidae. This study provides novel information on the reproductive biology of the Curculionidae.The female reproductive system, ovary structure and ultrastructure of Trypophloeus klimeschi (Coleoptera: Curculionidae: Scolytinae) were investigated using light microscopy, scanning electron microscopy, and transmission electron microscopy. Its female reproductive system is comprised of two ovaries (each ovary has two ovarioles), lateral oviducts, common oviduct, spermathecal sac, spermathecal pump, two accessory glands and bursa copulatrix. Well-developed endoplasmic reticulum can be clearly seen in the secretory cells of spermathecal sac. This species has telotrophic meroistic ovarioles that are comprised of terminal filament, tropharium, vitellarium and pedicel. The terminal filaments are simple; each is comprised of cellular peritoneal sheath. The presence of several clusters of nurse cells in the tropharium is indicative that its ovarioles conform to the transition stage. This indicates that there are at least two different types (transition stage and secondary stage) of ovarioles in Curculionidae.

Interaction of Epididymal Epithelia and their Secretions with Spermatozoa Supports Functional and Morphological Changes During Long-Term Storage in the Chinese Soft-Shelled Turtle (Pelodiscus sinensis).

Post-testicular maturation of spermatozoa is crucial for attaining the morphological and functional capabilities needed for successful fertilization. Epididymal epithelia offer a favorable environment for spermatozoa that are stored long term in the turtle epididymis; however, sperm-epithelial interactions during storage, which are enormously important for sperm functional and morphological maturation, are still largely unknown in turtles. The present study examined the epididymis during the sperm-storage period (November-April) in the Chinese soft-shelled turtle (Pelodiscus sinensis). Light and transmission electron microscopy were used to determine the cellular features of each epididymal segment (caput, corpus, and cauda) and their epithelial interactions with the spermatozoa. Spermatozoa were mainly located in the lumena of caput, corpus, and cauda epididymides. Numerous spermatozoa were bound to apical surfaces of the epithelia, and several were even embedded in the epithelial cytoplasm of the caput and corpus epididymides. No embedded spermatozoa were found in the cauda epididymis. In all epididymal segments, principal and clear cells showed the synthetic activity, evidenced by a well-developed endoplasmic reticulum network and high and low electron-dense secretory materials, respectively. Principal and clear cells in the caput and corpus segments showed embedded spermatozoa in electron-dense secretions and in the lipid droplets within the cytoplasm. No lysosomes were observed around the embedded spermatozoa. The lumena of the caput and corpus segments showed few apocrine and low electron density secretions. In the lumen of the cauda epididymidis, different secretions, such as holocrine with low and high electron density and their fragmentation, apocrine, and dictyosome, were found and are summarized. Altogether, sperm physical interactions with secretions either in the cytoplasm of epithelium or in the lumen may support the viability, morphological maintenance, and transfer of various proteins involved in long-term sperm storage in the turtle. This interaction could help us to understand the mechanisms of long-term sperm storage and provide more insights into the reproductive strategies of turtle sperm preservation.

Accessory glands of male reproductive system in Pseudochorthippus parallelus parallelus (Zetterstedt, 1821) (Orthoptera: Acrididae): A light and electron microscopic study.

The accessory glands of male reproductive system in insects play a significant role in the reproduction process by protecting sperm in spermatheca, preventing female to accept other males after mating and stimulating oviposition. The number, structure, and arrangement of the tubules of accessory glands can change from species to species. In this study, the accessory glands belonging the male reproductive system in Pseudochorthippus parallelus parallelus (Zetterstedt, 1821) (Orthoptera, Acrididae) were examined with stereomicroscope, light microscope, scanning (SEM), and transmission (TEM) electron microscopes at Gazi University, Faculty of Science in 2017-2019. P. parallelus parallelus is a widespread species that is located at the extending areas from Italy to the Northern Europe and also in Turkey. The accessory glands of P. parallelus parallelus' male reproductive system are composed of about 10 tubules. The tubules can be classified into two groups according to the thickness of their muscle tissues. Both groups have single layered epithelial cells with mitochondria, well-developed endoplasmic reticulum, spherical nucleus with electron dense chromatin, secretory vesicles and multivesicular bodies in their cytoplasm. In addition, apocrine type secretion is seen in epithelial cells.

Axonal Activation of the Unfolded Protein Response Promotes Axonal Regeneration Following Peripheral Nerve Injury

Adult mammalian peripheral neurons have an intrinsic regrowth capacity in response to axonal injury. The induction of calcium ion (Ca2+) oscillations at an injured site is critical for the regulation of regenerative responses. In polarized neurons, distal axonal segments contain a well-developed endoplasmic reticulum (ER) network that is responsible for Ca2+ homeostasis. Although these characteristics implicate the relevance among injury-induced Ca2+ dynamics, axonal ER-derived signaling, and regenerative responses propagated along the axons, the details are not fully understood. In the present study, we found that Ca2+ release from the axonal ER was accelerated in response to injury. Additionally, axonal injury-dependent Ca2+ release from the ER activated unfolded protein response (UPR) signaling at injured sites. Inhibition of axonal UPR signaling led to fragmentation of the axonal ER and disrupted growth cone formation, suggesting that activation of axonal UPR branches following axonal injury promotes regeneration via regulation of ER reconstruction and formation of growth cones. Our studies revealed that local activation of axonal UPR signaling by injury-induced Ca2+ release from the ER is critical for regeneration. These findings provide a new concept for the link between injury-induced signaling at a distant location and regulation of organelle and cytoskeletal formation in the orchestration of axonal regeneration.

Combined DOG1 and Mammaglobin Immunohistochemistry Is Comparable to ETV6-breakapart Analysis for Differentiating Between Papillary Cystic Variants of Acinic Cell Carcinoma and Mammary Analogue Secretory Carcinoma.

We investigated the reliability of combined DOG1 and mammaglobin immunohistochemistry compared with ETV6 fluorescence in situ hybridization (FISH) in the assessment of salivary tumors previously diagnosed as acinic cell carcinoma (ACC). Ultrastructural features of cases reclassified as mammary analogue secretory carcinoma (MASC) were assessed by transmission electron microscopy (TEM). Immunohistochemical (IHC) reactivity to DOG1 and mammaglobin was validated against FISH targeting the ETV6 gene in all 14 cases. Three cases with papillary cystic histomorphology previously diagnosed as ACC were revised to MASC. TEM features of the ETV6 rearrangement-positive MASC cases showed large numbers of secretory granules with extrusion into the intercellular spaces, well-developed endoplasmic reticulum, lipid-laden vacuoles, well-formed microvilli, and large lining cystic spaces. Combined DOG1 and mammaglobin immunohistochemistry is comparable to ETV6 -breakapart analysis for differentiating between papillary cystic variants of ACC and MASC.

Ultrastructural changes in the epidermis of petals of the sweet orange infected by Colletotrichum acutatum.

Postbloom fruit drop (PFD) is an important disease caused by the fungus Colletotrichum acutatum. PFD is characterised by the formation of necrotic lesions on the petals and stigmas of flowers as well as premature abscission of the fruit in Citrus spp. We compare the ultrastructure of the epidermis of uninoculated Citrus sinensis petals with that of petals inoculated with the fungus to understand the changes that occur upon C. acutatum infection. Healthy petals have a cuticle with parallel striations covering the uniseriate epidermis. This pattern consists of vacuolated parietal cells whose cytoplasm contains mitochondria, plastids with an undeveloped endomembrane system and a slightly dense stroma, a poorly developed rough endoplasmic reticulum, polysomes, few lipid droplets, and a nucleus positioned near the inner periclinal wall. In damaged regions, the cytoplasm of some cells is densely packed with well-developed endoplasmic reticulum, a large number of hyperactive dictyosomes, numerous mitochondria, and many lipid droplets. The plastids have an electron-dense stroma, starch grains, and a large amount of electron-dense lipid droplets, which can be released into vacuoles or the endoplasmic reticulum. Multivesicular bodies and myelin bodies are frequently observed in the vacuole, cytoplasm, and periplasmic space. Vesicles migrate through the cell wall and are involved in the deposition of cuticular material. In the later stages of infection, there is deposition of new cuticle layers in plaques. The outer periclinal walls can be thick. These observations indicate that epidermal cells respond to the pathogen, resulting in cuticular and parietal changes, which may limit further infection.

Trophic actions of oral vanadium and improved glycemia on the pancreatic beta-cell ultrastructure of streptozotocin-induced diabetic rats.

Oral vanadyl sulfate (vanadium) has potent hypoglycemic effects in diabetes animals, but data about its actions on pancreatic beta-cells (BC) ultrastructure is limited. Partial diabetic rats were treated with vanadium and insulin injection and their effects on BC ultrastructure are studied. Male rats were made diabetic with intravenous streptozotocin injection (STZ, 40 mg/kg). Animals were randomly divided to control (CD), vanadium-treated (1 mg/mL VOSO4 + 5H2O in base solution, VTD) and insulin-treated (80 U/kg/day NPH insulin injection, ITD) diabetic groups. Treatments started 10 days after STZ injection and terminated after 2 months. Intermittent tail blood samples were taken for measurements of blood glucose (BG) and plasma insulin (PI). Finally animals were sacrificed and pancreata prepared for assessments of BC ultrastructure, islets histology and insulin immunoreactivity (IIR). Vanadium decreased BG (P<0.0001), elevated the reduced PI (P<0.001), prevented islet atrophy and restored BC ultrastructure. Low BG seen during treatment in VTD and ITD only persisted in VTD after vanadium withdrawal. Hyperglycemia worsened in CD and repaired in ITD shortly after insulin withdrawal. CD islets were atrophied, had scattered IIR spots. BC had pyknotic nuclei, vacuolated cytoplasm and few tiny insulin secretory granules. VTD islets looked normal with compact centered IIR spots. BC had well-developed endoplasmic reticulum, many insulin secretory granules and mitochondria. ITD islet structure was slightly better than CD and BC had some immature insulin secretory granules. The trophic actions of vanadium in diabetic rats effectively renovated beta cell ultrastructure and prevented pancreatic islets atrophy, whereas the relief of glucotoxicity seen with insulin treatment could repair some beta cells and partially prevented islet atrophy.

Fine Structure of the Male Reproductive System and Reproductive Behavior of Lutzomyia longipalpis Sandflies (Diptera: Psychodidae: Phlebotominae)

BackgroundThe male reproductive system of insects can have several tissues responsible for the secretion of seminal fluid proteins (SFPs), such as testes, accessory glands, seminal vesicles, ejaculatory duct and ejaculatory bulb. The SFPs are transferred during mating and can induce several physiological and behavioral changes in females, such as increase in oviposition and decrease in sexual receptivity after copulation. The phlebotomine Lutzomyia longipalpis is the main vector of visceral leishmaniasis. Despite its medical importance, little is known about its reproductive biology. Here we present morphological aspects of the male L. longipalpis reproductive system by light, scanning and transmission electron microscopy, and compare the mating frequency of both virgin and previously mated females.ResultsThe male L. longipalpis reproductive system is comprised by a pair of oval-shaped testes linked to a seminal vesicle by vasa deferentia. It follows an ejaculatory duct with an ejaculatory pump (a large bulb enveloped by muscles and associated to tracheas). The terminal endings of the vasa deferentia are inserted into the seminal vesicle by invaginations of the seminal vesicle wall, which is composed by a single layer of gland cells, with well-developed endoplasmic reticulum profiles and secretion granules. Our data suggest that the seminal vesicle acts both as a spermatozoa reservoir and as an accessory gland. Mating experiments support this hypothesis, revealing a decrease in mating frequency after copulation that indicates the effect of putative SFPs.ConclusionUltrastructural features of the L. longipalpis male seminal vesicle indicated its possible role as an accessory gland. Behavioral observations revealed a reduction in mating frequency of copulated females. Together with transcriptome analyses from male sandfly reproductive organs identifying ESTs encoding orthologs of SFPs, these data indicate the presence of putative L. longipalpis SFPs reducing sexual mating frequency of copulated females.

Meningeal‐like organization of neural tissues in calanoid copepods (Crustacea)

Meninges, the connective tissue of the vertebrate central nervous system (CNS), have not been recognized in invertebrates. We describe the ultrastructure of the adult brain, antennules, and cord in five marine copepods: Calanus finmarchicus, Gaussia princeps, Bestiolina similis, Labidocera madurae, and Euchaeta rimana. In all of these locations we identified cell types with characteristics of the typical cells of vertebrate meninges and of their peripheral nervous system (PNS) connective tissue counterpart: fibroblasts, having flattened twisting processes with labyrinthine cavities communicating with the extracellular space, and macrophages, containing prominent lysosomes, well-developed endoplasmic reticulum, Golgi apparatus, and indented heterochromatin. The vertebrate distinction between electron-dense cells in the most external connective tissues (dura mater and epineurium) versus electron-lucent cells in the more internal connective tissues (pia-arachnoid and endoneurium-perineurium) was also found in the copepod CNS and PNS. Similar to the vertebrate organization, electron-dense cell networks penetrated from the outer layer (subcuticle) to surround inner substructures of the copepod nervous systems, and electron-lucent networks penetrated deeply from the brain and nerve surfaces to form intertwined associations with neural cells. Moreover, the association of these cells with basement membranes, glycocalyx, and fibrils of collagen in copepods conforms to a meningeal organization. The primary deviation from the vertebrate ultrastructural organization was the often tight investment of axons by the meningeal-like cells, with an intercalated basement membrane. Together, these data suggest that the tissues investing the copepod nervous system possess an organization that is analogous in many respects to that of vertebrate meninges.

New insight into the morphology of Eurytrema coelomaticum (Trematoda, Dicrocoeliidae) cercariae by light, scanning, and transmission electron microscopies

Eurytrema coelomaticum is a digenetic trematode that parasitizes the pancreatic ducts of ruminants. In the present study, the morphology of the cercariae was analyzed using light, scanning, and transmission electron microscopies. The size of the larvae was larger than that reported in the literature. An oral sucker with many papillae and an oral aperture in its center and a ventral sucker with few discrete papillae on the edge were observed. No stylet at the anterior end of the larval body and spines on the tegument of the end of the tail were observed. The cercariae had lateral penetration glands and (central) pre-acetabular glands, from which ducts lead to the anterior region and open into a small anterior pocket. The flame cells were located laterally and communicated with fine branches, converging to two lateral excretory collecting ducts that opened into an excretory bladder, centrally located at the posterior end of the body. The tegument presented an external layer loosely attached to the larval body, below which an amorphous syncytial outer layer with many mitochondrial profiles was observed. This region exhibited many secretions and released secretory granules, indicating intense secretory activity. The circular and longitudinal muscle layers were arranged in sequence below the outer layer. The outer layer was connected by cytoplasmic bridges crossing the muscular layers to the internal region of the tegument, where the cell body with nucleus was located. In the cercarial body, it was possible to observe a typical flame cell with the barrel region, where cilia and the internal and external ribbed regions were found. The somatic cells seem to have an active metabolism, with a well-developed endoplasmic reticulum, secretory granules, and evident nuclei. The results are discussed in the light of the biology and taxonomy of this species.

Thapsigargin down‐regulates protein levels of GRP78/BiP in INS‐1E cells

Pancreatic β-cells have a well-developed endoplasmic reticulum (ER) and express large amounts of chaperones and protein disulfide isomerases (PDI) to meet the high demand for synthesis of proteins. We have observed an unexpected decrease in chaperone protein level in the β-cell model INS-1E after exposure to the ER stress inducing agent thapsigargin. As these cells are a commonly used model for primary β-cells and has been shown to be vulnerable to ER stress, we hypothesize these cells are incapable of mounting a chaperone defense upon activation of ER stress. To investigate the chaperone expression during an ER stress response, induced by thapsigargin in INS-1E cells, we used quantitative mass spectrometry based proteomics. The results displayed a decrease of GRP78/BiP, PDIA3 and PDIA6. Decrease of GRP78/BiP was verified by Western blot and occurred in parallel with enhanced levels of p-eIF2α and CHOP. In contrast to INS-1E cells, GRP78/BiP was not decreased in MIN6 cell or rat and mouse islets after thapsigargin exposure. Investigation of the decreased protein levels of GRP78/BiP indicates that this is not a consequence of reduced mRNA expression. Rather the reduction results from the combined effect of reduced protein synthesis and enhanced proteosomal degradation and possibly also degradation via autophagy. Induction of ER stress with thapsigargin leads to lower protein levels of GRP78/BiP, PDIA3 and PDIA6 in INS-1E cells which may contribute to the susceptibility of ER stress in this β-cell model.

Micromorphology and ultrastructure of the floral nectaries of Polemonium caeruleum L. (Polemoniaceae)

The Polemoniaceae family forms flowers diverse in the terms of pollination methods and nectar types. The micromorphology of the nectary surface and the tissue structures as well as the ultrastructure of the cells of the floral nectaries in Polemonium caeruleum L. were examined using light, scanning and transmission electron microscopy. A bowl-shaped nectary, detached from the ovary, grows at its base. Its contour shows folds with depressions in the places where the stamens grow, forming five-lobed disc (synapomorphic character). Nectar is secreted through modified anomocytic stomata, which are formed in the epidermis covering the tip and the lateral wall of the projection located between the staminal filaments. The undulate nectary consists of a single-layered epidermis and three to nine layers of parenchymal cells. The cells of the nectary contain a dense cytoplasm, numerous plastids with an osmophilic stroma and starch grains, well-developed endoplasmic reticulum, as well as a large number of mitochondria interacting with the Golgi bodies. The ultrastructure of nectary cells indicates the granulocrine secretion mechanism and diversified transport of nectar.

개조개, Saxidomus purpuratus 소화맹낭의 미세구조

개조개 소화맹낭의 해부학적 구조와 미세구조를 광학 및 전자현미경을 이용하여 기재하였다. 개조개는 한국 여수연안에서 2010년 5월에 채집하였다. 소화맹낭은 진한 녹색으로 생식소 위쪽에 위치하며, 일차소관으로 위와 연결되어 있었다. 소화맹낭은 다수의 소화선세관들로 구성되며, 각각의 소화선세관은 단층 상피층으로 호염기성세포와 소화세포들로 이루어져 있었다. 호염기성세포는 원주형으로 소화세포에 비해 전자밀도가 높았다. 세포질에는 잘 발달된 조면소포체, 관상의 미토콘드리아, 골지체 및 전자밀도가 높고 막을 가진 분비과립들을 함유하고 있었다. 소화세포는 원주형이며, 자유면에는 미세융모가 발달되어 있었다. 세포질 상부에서는 음소포, 용해소체 및 미토콘드리아가 관찰되었다. 본 연구에서 이러한 결과는 소화선세관의 호염기성세포와 소화세포는 각각 세포외 소화와 세포내 소화에 적당하게 분화되었음을 의미한다. The anatomy and ultrastructure of the digestive diverticulum of Saxidomus purpuratus were described using light and electron microscopy. The digestive diverticulum of dark green color was situated on the gonad and connected to stomach by a primary duct. Digestive diverticulum is composed of numerous digestive tubules. The epithelial layer of digestive tubule, which is simple, is composed of basophilic cells and digestive cells. Basophilic cells are columnar in shape, and the electron density is higher than that of the digestive cell. The cytoplasm has a well-developed endoplasmic reticulum, tubular mitochondria, Golgi complex and membrane-bounded granules of high electron density. Digestive cells are columnar in shape, with development of microvilli on the free surface. Pinocytic vasicles, lysosomes and numerous mitochondria were observed in the apical cytoplasm of digestive cells. The results of this study suggest that basophilic cells and digestive cells in the digestive tubule are specialized in the extracellular and intracellular digestions, respectively.

A novel triazolic naphthofuranquinone induces autophagy in reservosomes and impairment of mitosis in Trypanosoma cruzi

Chagas' disease, caused by the protozoan Trypanosoma cruzi, represents a serious health problem in Latin America, and the available chemotherapy, which is based on 2 nitro-derivatives, is not satisfactory. In folk medicine, natural products including naphthoquinones have been employed for the treatment of different parasitic diseases. In the pursuit of alternative drugs for Chagas' disease, we investigated the mechanism of action of the triazolic naphthoquinone (TN; 2,2-dimethyl-3-(4-phenyl-1H-1,2,3-triazol-1-yl)-2,3-dihydronaphtho[1,2-b]furan-4,5-dione), which is the most active compound against T. cruzi trypomastigotes among a series of naphthofuranquinones. TN was active against the 3 parasite forms producing a dose-dependent inhibitory effect. In epimastigotes, TN induced reservosome disruption, flagellar blebbing, Golgi disorganization, the presence of cytosolic concentric membrane structures and abnormal multiflagellar parasites. The treatment also led to the appearance of well-developed endoplasmic reticulum profiles surrounding organelles that associated with an increase in monodansylcadaverine labelling, suggesting autophagy as part of the TN mechanism of action. Interestingly, no ultrastructural damage was detected in the mitochondria of naphthoquinone-treated epimastigotes. Flow cytometric analysis demonstrated an impairment of mitosis, an increase in ROS production and the maintenance of mitochondrial membrane potential. TN could be a good starting point in the investigation of a chemotherapeutic approach for the treatment of Chagas' disease.

Binding of activating transcription factor 6 to the A5/Core of the rat insulin II gene promoter does not mediate its transcriptional repression

Pancreatic β-cells have a well-developed endoplasmic reticulum due to their highly specialized secretory function to produce insulin in response to glucose and nutrients. It has been previously reported that overexpression of activating transcription factor 6 (ATF6) reduces insulin gene expression in part via upregulation of small heterodimer partner. In this study, we investigated whether ATF6 directly binds to the insulin gene promoter, and whether its direct binding represses insulin gene promoter activity. A bioinformatics analysis identified a putative ATF6 binding site in the A5/Core region of the rat insulin II gene promoter. Direct binding of ATF6 was confirmed using several approaches. Electrophoretic mobility shift assays in nuclear extracts from MCF7 cells, isolated rat islets and insulin-secreting HIT-T15 cells showed ATF6 binding to the native A5/Core of the rat insulin II gene promoter. Antibody-mediated supershift analyses revealed the presence of both ATF6 isoforms, ATF6α and ATF6β, in the complex. Chromatin immunoprecipitation assays confirmed the binding of ATF6α and ATF6β to a region encompassing the A5/Core of the rat insulin II gene promoter in isolated rat islets. Overexpression of the active (cleaved) fragment of ATF6α, but not ATF6β, inhibited the activity of an insulin promoter-reporter by 50%. However, the inhibitory effect of ATF6α was insensitive to mutational inactivation or deletion of the A5/Core. Therefore, although ATF6 binds directly to the A5/Core of the rat insulin II gene promoter, this direct binding does not appear to contribute to its repressive activity.

Ultrastructural and Immunohistochemical Analysis of Intestinal Myofibroblasts During the Early Organogenesis of the Human Small Intestine

Intestinal myofibroblasts (IMFs), also known as pericryptal fibroblasts, are found at the basement membrane of the intestinal epithelium. They are characterized by well-developed endoplasmic reticulum, cytoplasmic fibers, and fibrous extensions called fibronexi. IMFs have structural features in common both with fibroblasts and smooth cells. Vimentin, desmin, and α-smooth-muscle actin (α-SM) are markers commonly used to discriminate between IMFs and smooth muscle cells. Immunohistochemical studies have shown that, when α-SM and vimentin are positive in both IMFs and smooth muscle cells, desmin is negative in IMFs but positive in smooth muscle cells. In the adult intestine, IMFs play an important role in various functions, especially in tissue repair and scar formation during wound healing. In the embryonic intestine, however, wound healing does not occur, and to date, no studies have investigated the first appearance and subsequent evolution of IMFs. In this study, we have examined the human small intestine in embryos at 7, 9, and 11 weeks of development by ultrastructural and immunohistochemical analysis to shed light on the formation of IMFs during these early phases of organogenesis. At 7 weeks, the embryonic mesenchymal cells are similar to proto-myofibroblasts and may be the precursors of the IMFs detected at 9 weeks and more abundantly at 11 weeks by immunohistochemistry. These IMFs seem to mediate information flow between the epithelium and the mesenchyme and thus contribute to the development of the small intestine.

Ultrastructural study of naturally occuring ovine pulmonary adenocarcinoma in Fars province, Iran

Summary Ovine pulmonary adenocarcinoma (OPA) was studied in the lungs of 15 naturally-affected sheep (9 with classical and 6 atypical lesions) by transmission electron microscopy. Two lung samples from normal sheep were used to develop the ultrastructural criteria. Pathologic lesions consisted of focal-multifocal to coalescent nodules or masses in the cranioventral or diaphragmatic lobes. Ultrastructural characteristics of tumor cells in the alveoli and bronchioles allowed categorization of these cells into three groups; neoplastic alveolar type (AT) П cells in the alveoli contained numerous cytoplasmic lamellar bodies, well developed rough endoplasmic reticulum and glycogen particles; neoplastic clara cells in the bronchioles contained apical electron-dense granules and well-developed smooth endoplasmic reticulum; undifferentiated tumor cells in the alveoli and bronchioles lacked characteristic lamellar bodies or electron-dense granules. Neither complete virions nor viral inclusions were seen in the neoplastic cells. In the present study, there were no ultrastructural differences in the neoplastic cells between classical and atypical forms. It seems that type II pneumocytes and clara cells are the origin of the neoplasia in the alveoli and bronchioles, respectively.