Dilated Cisternae Research Articles

Maternal exposure to low-dose bisphenol A and its potential neurotoxic impact on male pups: A histological, immunohistochemical, and ultrastructural study

BackgroundBisphenol A (BPA) is a widely used chemical with a harmful effect on animal and human. The neonatal and juvenile period is a highly risky neurodevelopmental period. AimThis study aimed to determine how male albino rat pups’ cerebral cortex was altered by low doses of BPA given to mothers and the role of the oxidative stress. MethodsThirty pregnant rats were randomly split into three equal groups, negative control, and positive control: received 1 cc of corn oil once a day through gastric tube and BPA treated: a dose of 200 µg/kg/day (dissolved in 1 cc corn oil). The male rat pups of each group were sacrificed at 1 week, 3 weeks and 6 weeks. The cerebra were then separated from the brain for histological and biochemical studies. ResultsRats administered BPA had raised levels of lipid peroxidation marker (MDA), lower levels of enzymatic antioxidants (SOD and CAT) with decreased body, cerebral weights, and decreased levels of non-enzymatic antioxidant defense (GSH). Histo-pathologically, shrunken pyramidal cells with congested blood vessels appeared. GFAP displayed increased number of positive immune-reactive astrocytes with high statistically significant increase in the area % in BPA treated group when compared to the control groups, on contrary to MBP. Semi-thin and ultra-thin BPA-sections revealed degenerative changes in myelinated axons with tiny nucleus and broken nuclear membranes. Lysosomes, dilated endoplasmic reticulum cisternae with noticeable increase in unmyelinated nerve fibers were also observed. ConclusionThe structure of the developing cerebral cortex is negatively impacted by BPA due to oxidative stress.

Echinops Asteraceae extract guards against malathion-induced liver damage via minimizing oxidative stress, inflammation, and apoptosis

Malathion (MAL) is one of the highly toxic organophosphorus (OP) compounds that induces hepatotoxicity. Echinops. ritro leaves extract (ERLE) is traditionally used in the treatment of bacterial/fungal infections. This study's goal was to investigate the potential of extracts from ERLE against hepatotoxicity induced by MAL in male albino rats. Four equal groups of forty mature male albino rats were created: The rats in the first group used as a control. The second group of rats received ERLE orally. The third group received MAL. ERLE and MAL were administered to the fourth group of rats. Six-week treatment groups were conducted. Using lipid peroxidation indicators [malondialdehyde (MDA), alanine aminotransferase (ALT), aspartate aminotransferase (AST)], oxidative stress markers [catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GPx)], apoptotic markers [Bcl-2 & caspase-3] and tumor necrosis factor alpha (TNF-α). Rats treated with MAL underwent a significant increase on MDA, ALT, AST, caspase-3 and TNF-α marker with a significant decrease in antioxidant markers [CAT, SOD, GPx] and Bcl-2. Histologically, MAL-treated group's liver sections displayed damaged hepatocytes with collapsed portions, pyknotic nuclei, vacuolated cytoplasm, and congested central veins. Ultra structurally, rat livers treated with MAL showed dilated cisternae of endoplasmic reticulum, swollen mitochondria with disrupted cristae, nuclei with disrupted chromatin content, multiple lysosomes, multiple vacuolations and a disrupted blood sinusoid. With rats treated with ERLE, these alterations were essentially non-existent. It is possible to conclude that ERLE protects against MAL hepatotoxicity, and that this protection is related, at least in part, to its antioxidant activities.

Protective effects of Urtica dioica on the cerebral cortex damage induced by Potassium bromate in adult male albino rats

ABSTRACT Potassium bromate is used in cheese production, beer making and is also used in pharmaceutical and cosmetic. It is a proven carcinogen as it is a strong oxidizing agent that generates free radicals during xenobiotic metabolism. Urtica dioica (Ud) (from the plants’ family of Urticaceae) is a plant that has long been used as a medicinal plant in many parts of the world. It has been shown to have anti-inflammatory, antioxidant and immunosuppressive properties. So, this study aimed to clarify the effect of Potassium bromate on the histological structure of cerebral cortex of adult male albino rats, evaluate the possible protective role of Urtica dioica. Thirty adult healthy male albino rats were divided into three groups; group I (Control group), group II (KBrO3 treated group). Group III (KBrO3 and Urtica dioica treated group).At the end of the experiment, rats in all groups were anesthetized and specimens were processed for light and electron microscope. Morphometric and statistical analyses were also performed. Nerve cells of the treated group showed irregular contours, dark nuclei, irregular nuclear envelopes, dilated RER cisternae, and mitochondria with ruptured cristae. Vacuolated neuropil was also observed. Immunohistochemically, stained sections for GFAP showed strong positive reaction in the processes of astrocytes. Recovery group showed revealed nearly the same as the histological picture as the control group. In conclusion, potassium bromate induces degenerative effects on neurons of cerebral cortex and urtica dioica provide an important neuroprotective effects against these damaging impacts through their antioxidant properties.

The presence of Mott cells in the lymph nodes of rats with experimental autoimmune encephalomyelitis.

Mott cells are plasma cells that have multiple spherical Russell bodies packed in their cytoplasm. Russell bodies are dilated endoplasmic reticulum cisternae filled with aggregates of immunoglobulins that are neither secreted nor degraded. Mott cells were observed in our study by light and electron microscope in the lymph nodes of rats with experimental autoimmune encephalomyelitis (EAE), an animal model of multiple sclerosis. Mott cells were detected on hematoxylin and eosin (HE)-stained lymph node sections as vacuolated cells with eccentrically positioned nuclei and large number of faint blue spherical inclusions in the cytoplasm. Electron microscopic investigation revealed the presence of Russell bodies of the "medusa" form inside Mott cells in lymph node ultra-thin sections of EAE animals. Mott cells expressed the plasma cell marker CD138 and either kappa or lambda immunoglobulin light chains, indicating their origin from polyclonally activated B cells. Finally, Mott cells were associated with active EAE, as they were not found in the lymph nodes of EAE-resistant Albino Oxford rats. The presence of Russell bodies implies an excessive production of immunoglobulins in EAE, thus further emphasizing the role of B cells, and among them Mott cells, in the pathogenesis of this animal model of multiple sclerosis.

Fallopia japonica and Fallopia × bohemica extracts cause ultrastructural and biochemical changes in root tips of radish seedlings.

Japanese knotweed (Fallopia japonica) and Bohemian knotweed (Fallopia × bohemica) are invasive plants that use allelopathy as an additional mechanism for colonization of the new habitat. Allelochemicals affect the growth of roots of neighboring plants. In the present study, we analyze the early changes associated with the inhibited root growth of radish seedlings exposed to aqueous extracts of knotweed rhizomes for 3 days. Here, we show that cells in the root cap treated with the knotweed extracts exhibited reduced cell length and displayed several ultrastructural changes, including the increased abundance of dilated ER cisternae filled with electron-dense material (ER bodies) and the accumulation of dense inclusions. Moreover, mitochondrial damage was exhibited in the root cap and the meristem zone compared to the non-treated radish seedlings. Furthermore, malfunction of the intracellular redox balance system was detected as the increased total antioxidative capacity. We also detected increased metacaspase-like proteolytic activities and, in the case of 10% extract of F. japonica, increased caspase-like proteolytic activities. These ultrastructural and biochemical effects could be the reason for the more than 60% shorter root length of treated radish seedlings compared to controls.

Loss of CAPS2/Cadps2 leads to exocrine pancreatic cell injury and intracellular accumulation of secretory granules in mice.

The type 2 Ca2+-dependent activator protein for secretion (CAPS2/CADPS2) regulates dense-core vesicle trafficking and exocytosis and is involved in the regulated release of catecholamines, peptidergic hormones, and neuromodulators. CAPS2 is expressed in the pancreatic exocrine acinar cells that produce and secrete digestive enzymes. However, the functional role of CAPS2 in vesicular trafficking and/or exocytosis of non-regulatory proteins in the exocrine pancreas remains to be determined. Here, we analyzed the morpho-pathological indicators of the pancreatic exocrine pathway in Cadps2-deficient mouse models using histochemistry, biochemistry, and electron microscopy. We used whole exosome sequencing to identify CADPS2 variants in patients with chronic pancreatitis (CP). Caps2/Cadps2-knockout (KO) mice exhibited morphophysiological abnormalities in the exocrine pancreas, including excessive accumulation of secretory granules (zymogen granules) and their amylase content in the cytoplasm, deterioration of the fine intracellular membrane structures (disorganized rough endoplasmic reticulum, dilated Golgi cisternae, and the appearance of empty vesicles and autophagic-like vacuoles), as well as exocrine pancreatic cell injury, including acinar cell atrophy, increased fibrosis, and inflammatory cell infiltration. Pancreas-specific Cadps2 conditional KO mice exhibited pathological abnormalities in the exocrine pancreas similar to the global Cadps2 KO mice, indicating that these phenotypes were caused either directly or indirectly by CAPS2 deficiency in the pancreas. Furthermore, we identified a rare variant in the exon3 coding region of CADPS2 in a non-alcoholic patient with CP and showed that Cadps2-dex3 mice lacking CAPS2 exon3 exhibited symptoms similar to those exhibited by the Cadps2 KO and cKO mice. These results suggest that CAPS2 is critical for the proper functioning of the pancreatic exocrine pathway, and its deficiency is associated with a risk of pancreatic acinar cell pathology.

The possible protective role of pimpinella anisum oil versus selenium on aspartame induced changes in rat cerebellar cortex: histological, immunohistochemical and electron microscopic study

ABSTRACT Aspartame (ASP) is an artificial sweeter. Chronic use of ASP has a harmful effect on cerebellar cortex. Anisum oil and selenium (SE) are antioxidant substances. Therefore, the present study was performed to study the possible protective role of anisum oil versus selenium on aspartame-induced changes in rat cerebellar cortex. Rats were divided into four main groups. Group I (Control group). Group II received 250 mg/kg/day aspartame once daily for 2 months. Group III received 0.5 ml/kg/day anisum 2 h before aspartame administration. Group IV received 0.5 mg/kg/day selenium 2 h before aspartame administration. The administration of Asp for 2 months (group II) resulted in cerebellar histopathological changes in the form of deformed Purkinje and granule cells. Ultrastructurally, Purkinje cells had irregular nuclei, dilated cisternae of rough endoplasmic reticulum, dilated saccules of Golgi apparatus, mitochondria with destroyed cristae. In addition, granule cells appeared shrunken with irregular nuclei. Aspartame and anisum oil treated group (group III) showed partial improvement. Examination of ASP and SE treated group (group IV) showed that cerebellar cortex was nearly similar to control. In conclusion, Anisum oil and selenium could protect against ASP-induced cerebellar damage. The protective effect of selenium is better than anisum oil.

CREB3L1 and CREB3L2 control Golgi remodelling during decidualization of endometrial stromal cells.

Upon progesterone stimulation, Endometrial Stromal Cells (EnSCs) undergo a differentiation program into secretory cells (decidualization) to release in abundance factors crucial for embryo implantation. We previously demonstrated that decidualization requires massive reshaping of the secretory pathway and, in particular, of the Golgi complex. To decipher the underlying mechanisms, we performed a time-course transcriptomic analysis of in vitro decidualizing EnSC. Pathway analysis shows that Gene Ontology terms associated with vesicular trafficking and early secretory pathway compartments are the most represented among those enriched for upregulated genes. Among these, we identified a cluster of co-regulated genes that share CREB3L1 and CREB3L2 binding elements in their promoter regions. Indeed, both CREB3L1 and CREB3L2 transcription factors are up-regulated during decidualization. Simultaneous downregulation of CREB3L1 and CREB3L2 impairs Golgi enlargement, and causes dramatic changes in decidualizing EnSC, including Golgi fragmentation, collagen accumulation in dilated Endoplasmic Reticulum cisternae, and overall decreased protein secretion. Thus, both CREB3L1 and CREB3L2 are required for Golgi reshaping and efficient protein secretion, and, as such, for successful decidualization.

Diving in the big blue: Atypical Mott cells in rapidly fatal primary amyloidosis.

A 75-year-old man with no previous medical history presented at the emergency unit with asthenia, anorexia, weight loss, and progressively increasing dyspnea revealing acute heart failure (N-terminal pro b-type natriuretic peptide:16 667 ng/L; troponemia: 47 ng/L). Echocardiography showed a granular appearance of the myocardium and interventricular septal hypertrophy, suggestive of amyloidosis. Chemistry disclosed high levels of immunoglobulin kappa light chains at 1240 mg/L (lambda light chains 14 mg/L; κ/λ ratio 86) and a detectable but unquantifiable IgA κ peak. Myocardial biopsies pathology showed an endocardium thickened by fibrosis. Within the myocardium, the presence of interstitial fibrillary deposits stained by Congo red and birefringent in polarized light confirmed the diagnosis of cardiac amyloid light-chain (AL) amyloidosis. The search for other organ damage disclosed vegetative involvement with orthostatic hypotension, icteric cholestasis (bilirubin 51.8 mg/L), undocumented digestive involvement (diarrhea with evocative endoscopic lesions), and glomerulonephritis (urinary proteinuria 4 g/24 h with 3 g/24 h albumin and 1 g/24 h immunoglobulin light chains). A bone marrow aspirate displayed 17% of plasma cells. Interestingly, a particular pattern of huge atypical Mott cells was identified (Figure 1A,B). The latter had so many cytoplasmic inclusions that the cytoplasm, and the nucleus could not be discriminated. These atypical cells are however plasma cells with an endoplasmic reticulum loaded with immunoglobulins and should be integrated as such in cell counts. Classical plasma cells and typical Mott cells with a few Russel bodies were also present (Figure 1C) indicating a continuum in the shape of the diseased plasma cells. Treatment with cyclophosphamide–bortezomib–dexamethasone-daratumumab was initiated in emergency. Unfortunately, the patient died 3 days later, of multiple organ failure. No additional septic or toxic causes could be identified. Mott cells are activated plasma cells containing intracytoplasmic inclusions with a “bunch of grapes” appearance. These inclusions, called Russell bodies, are dilated cisternae of the endoplasmic reticulum containing condensed immunoglobulins. They were identified in 1901 by the surgeon F.W Mott in the brains of monkeys with trypanosomiasis. Mott cells have been described in various pathological conditions: reactive plasmacytosis, monoclonal gammopathies, high grade B-cell lymphomas, Wiskott-Aldrich syndrome, and von Recklinghausen neurofibromatosis. AL amyloidosis is caused by clonal expansion of plasma cells producing misfolded immunoglobulin light chains responsible for the deposition of insoluble fibrillar proteins in many organs. Those most frequently affected are the kidneys, heart, liver, and peripheral nervous system. AL amyloidosis is a rare disease, and its diagnosis is often delayed. Although associated with a poor prognosis, AL amyloidosis presents a heterogeneous clinical expression and evolution. To our knowledge, the presence of Mott cells has rarely been described in this pathology. KR, ME, and BLC wrote the manuscript. KR, ME, and BLC performed biological analyses. CS treated the patient. All authors critically reviewed the manuscript. The authors have no conflict of interest to disclose. The authors received no specific funding for this work. The research was conducted in accordance with the principles embodied in the Declaration of Helsinki and in accordance with local statutory requirements.

Effect of ozone versus naringin on testicular injury in experimentally induced ulcerative colitis in adult male albino rats

ABSTRACT Testicular dysfunction is caused by the continuous inflammation and oxidative stress that are present at the local site in ulcerative colitis (UC) spreading to the testes via systemic circulation. The influence of ozone and naringine on colitis-mediated testicular dysfunction was investigated in this study. Forty-eight adult male rats were divided into four groups: I control group, II dextran sodium sulfate (DSS) UC-induced group, III DSS+naringine, and IV DSS+ozone groups. UC was induced in groups II, III, and IV using 0.1 ml of 4% DSS in their drinking water per day for 6 days by gastric gavage. All animals were sacrificed 45 days from the start. Blood samples were obtained to estimate serum testosterone hormone. Testicular tissues were processed for measurement of tissue malondialdehyde (MDA) and examined by light and electron microscopes. Ultrastructurally, group II revealed a relatively thick basement membrane enveloping the seminiferous tubule. Sertoli cell cytoplasm appears rarified with wide intracellular spaces, vacuoles, and multiple lysosomes; distorted spermatogonia with electron dense nuclei and cytoplasm; and primary spermatocytes with small nuclei and electron dense cytoplasm. Abnormal sperm profiles were visible in middle pieces, mid, principle, and end pieces that were markedly affected with disorganization of axoneme and outer dense fibers. Leydig cells revealed dilated cisternae of smooth endoplasmic reticulum. Morphometric and statistical analyses were performed. Group III showed some improvement; however, group IV showed more improvement. The results indicated that ozone caused marked improvement than naringine against UC-induced testicular damage via their antioxidant and anti-inflammatory properties.

Effect of <i>Ginkgo biloba</i> extract on Histomorphometric changes of Hippocampus against Fluoride Toxicity in Wistar Rats

Prolonged ingestion of fluoride leads to the pathogenesis known as fluorosis. Fluoride exposure leads to neuro degenerative changes such as reduction in the neuronal cell size and number. Exposure to fluoride showed neurodegenerative changes like shrunken neurons, increased in folding of the nuclear membrane, mitochondrial alterations, and dilated rough endoplasmic reticulum cisternae and clusters of vesicles near the Golgi bodies. Present study is to explore the histomorphometric changes of the hippocampus of fluoride induced brain and treated with Ginkgo biloba Extract (GBE). Thirty number (30) adult male Wistar rats were randomly separated into 5 Groups (n=6). Group1 (Control) supplemented with water, 2 to 5 Groups were supplemented with 100ppm of sodium fluoride for 30 days, while the Groups 3, 4, and 5 were Ginkgo biloba extract treated at 50mg/ kg, 100mg/kg and 200mg/kg body weight for 15 days, after sodium fluoride treatment, results showed that, 200mg/kg GBE provided effective and complete protection against fluoride toxicity. From the present study concluded that 50, 100 and 200 mg/kg GBE was significantly attenuated the fluoride induced toxicity in a dose depending manner. Present histomorphometrical study showed delayed neuronal death in the CA1 and CA3 regions of hippocampus of fluoride group that was significantly attenuated by GBE (50,100 and 200 mg/kg) in a dose dependant manner.

Benzo[ghi]perylene induces cellular dormancy signaling and endoplasmic reticulum stress in NL-20 human bronchial epithelial cells

Benzo[ghi]perylene (BghiP) is produced by the incomplete combustion of gasoline and it is a marker of high vehicular flow in big cities. Nowadays, it is known that BghiP functions as ligand for the aryl hydrocarbon receptor (AhR), which can cause several molecular responses. For this reason, the aim of the present study was to assess the in vitro effects of the exposure to BghiP, specifically, the induction of cellular dormancy and endoplasmic reticulum stress (ER stress) in NL-20 human cells. Our results proved that a 24 h exposure of BghiP, increased the expression of NR2F1 (p < 0.05). NR2F1 is the main activator of cell dormancy, therefore, we analyzed the expression of its target genes SOX9 and p27 showing an increase of the transcripts (p < 0.05), suggesting a pathway that could produce a cell cycle arrest. Interestingly, this effect was only observed with BghiP exposure, and not with a classic AhR ligand: benzo[a]pyrene. Moreover, in the presence of the AhR antagonist, CH223191, or when the expression of AhR was knock-down using dsiRNAs, the cellular dormancy signaling pathway was blocked. Morphological and ultrastructure analysis demonstrated that BghiP also induces ER stress, characterized by the dilated ER cisternae and the overexpression of PERK and CHOP genes (p < 0.05). Moreover, the halt of cell proliferation and the ER stress are both associated to the increase of pro-inflammatory cytokines (IL-6 and IL-8) and the cell survival in response to microenvironmental cues. These responses induced by BghiP on bronchial cells open new horizons on the research of other biological effects induced by environmental pollutants.

Isolation and morphological characterization of equine mesenchymal stem cells from harvested adipose tissue and bone marrow and stably transfected with green fluorescent protein.

To characterize the ultrastructure of mesenchymal stem cells (MSCs) that were harvested from the adipose tissue (AT-MSCs) and bone marrow (BM-MSCs) of horses and transfected with green fluorescent protein. MSCs from adipose tissue and bone marrow of 6 adult female Hispano-Bretón horses. Harvested equine MSCs were cultivated and transfected with green fluorescent protein, and the immunophenotypes of the MSCs were characterized by use of anti-CD90 and anti-CD105 monoclonal antibodies. When stable transfection of MSCs was achieved, the morphological and ultrastructural characteristics of transfected and nontransfected AT-MSCs and BM-MSCs were compared with electron microscopy. The protocols for transfection and subsequent isolation of transfected cells with use of G418 were suitable for obtaining transfected MSCs. Transfection efficiency was 5% in AT-MSCs and 4% in BM-MSCs. Characterization of transfected and nontransfected MSCs revealed that they share immunocytochemical and morphological profiles. Expression of CD90 was significantly higher for transfected versus nontransfected AT-MSCs (97% vs 92%). Expression of CD105 was significantly lower for transfected versus nontransfected BM-MSCs (85% vs 94%). Transfected BM-MSCs had differences in organelles, compared with the other cell types, specifically including most commonly the rough endoplasmic reticulum with dilated cisternae and mitochondria. These findings contribute to the knowledge base of the characteristics of equine AT-MSCs and BM-MSCs and of transfected versus nontransfected equine MSCs. The data provided a valuable starting point for researchers wishing to further study the morphological characteristics of equine MSCs.

Abstract 2032: Vitamin B12 augments Ca2+ dependent cytotoxicity of diethyldithiocarbamate through excessive ER-vacuolization, oxidative stress and mitochondrial depolarization in MCF-7 cells in culture

Abstract Coordinated organization of intracellular organelles into functional networks is vital for cells. Mitochondrial and endoplasmic reticulum (ER) interaction in regulation of cell Ca2+ homeostasis is crucial for cell death and survival. Function of these intracellular organelles involved in two-way regulation of the status of major cell metabolites including protein, lipid exchange and intracellular Ca2+ homeostasis. Disulfiram and its derivatives recently attracted attention as a potential anticancer treatment (Kona et al., 2011). Combined use of hydroxycobalamin (vitamin B12b) and diethyldithiocarbamate (DDTC) demonstrated synergistic effect (combination index=0.25) and enhanced the cytotoxic effect of DDTC in number of transformed cell lines in culture (Solovieva et al., 2020). In the present work using MCF-7, human breast cancer cells in culture we demonstrate that combined action of DDTC and B12b resulted in induction of UPRER manifested by dilation of ER cisterns, increased level of polyubiquitinated proteins, and increased expression of CHOP. Confocal microscopy revealed disorganization of mitochondrial network and ER and treatment of these cells with B12b/DDTC and accompanied by mitochondrial depolarization (~50% after 6 h of exposure to drugs). Treatments also induced changes in morphology of mitochondrial network, and within 6 h of exposure, mitochondria demonstrated large amplitude swelling, characteristic feature of mitochondrial permeability transition pore opening. Drug exposure induced mitochondrial oxidative stress and increase in superoxide anion level (monitored with MitoSOX) and increased content of mitochondrial Ca2+ (xRhod-1) which almost doubled following treatment with B12b/DDTC. Short-time exposure of MCF-7 cells to mixture of drugs in Ca2+-free incubation media (2.5 mM EGTA) resulted in generation of significantly smaller size of vacuoles as compared with control. Similarly, pre-loading of cells with intracellular Ca2+ chelator (BAPTA, 5 µM, 30 min, 37oC) or inhibition of IP3 receptor(s) with 2-APB, delayed the drug-induced vacuolization. Taken together, our data indicate that the mechanism of B12b/DDTC-induced cytotoxicity in human MCF-7 cancer cell lines involves mitochondrial oxidative stress, activation of MPT pore and Ca2+ overload. Support of Funding Information: Russian Science Foundation № 19-75-20145 Citation Format: Marina E. Solovieva. Vitamin B12 augments Ca2+ dependent cytotoxicity of diethyldithiocarbamate through excessive ER-vacuolization, oxidative stress and mitochondrial depolarization in MCF-7 cells in culture [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2032.

Ultrastructural evidence for the participation of muscle cells in the formation of extracellular matrices in aporocotylid blood flukes (Digenea)

The muscle cells and extracellular matrices (ECMs) of two teleost-infecting blood flukes belonging to distinct evolutionary lineages of the Aporocotylidae (Digenea) were examined using Transmission Electron Microscopy. Four morphotypes of muscle cells were found in the freshwater species Sanguinicola sp., but were considered to be various developmental stages of a single cisternic type. In the marine species Aporocotyle simplex, three types of muscle cells were apparent, one of which is cisternic. The first ultrastructural evidence is presented for the exocytosis of the moderately dense contents of dilated cisternae of cisternic muscle cells into the extracellular space in both Sanguinicola sp. and A. simplex. The basal matrices of aporocotylids support various types of epithelia. In Sanguinicola sp., beneath the distal tegumental cytoplasm, there is a thin lamina densa, whereas the intestinal epithelium is supported by a lamina reticularis. In A. simplex, both a thin lamina densa and a thick lamina reticularis underlie the distal cytoplasm of the tegument and are present at the periphery of the ovary, but beneath the epithelial lining of the caeca and both genital and excretory ducts there is only a lamina reticularis. Significant variation in the development and amount of the ECM in marine and freshwater aporocotylids is described, since A. simplex has a much better developed ECM than occurs in Sanguinicola sp. Moreover, thin myofilaments of muscle fibres participate in the ECM formation in A. simplex and represent its dominant component. The presence of two mechanisms for ECM formation in A. simplex, as opposed to a single mechanism in Sanguinicola sp., may represent further evidence for the affiliation of these two taxa to divergent evolutionary lineages. The data presented are discussed in relation to available information on these structures in other neodermatan groups.

Light and electron microscopic study on the possible protective effect of pomegranate peel extract on the pituitary- thyroid axis exposed to monosodium glutamate in adult male albino rats

Background: Monosodium glutamate is one of our new high-tech food additives. Nowadays, use of natural antioxidant as pomegranate peel extract has become an attractive therapeutic strategy for reducing the risk of disease.Aim of work: this study was designed to assess the effects of MSG on the pituitary thyroid axis and the possible protective role of pomegranate peel extract.Material & methods: Forty adult male albino rats were divided into three groups. Group I: (control) that was subdivided into; subgroups I A & I B . Group II (MSG treated) received MSG at a dose of (6mg/g/day). Group III (MSG & Pomegranate peel treated) received MSG & PPE as same dose as subgroup I B and group II. At the end of the experiment, pituitary and thyroid glands were dissected out and processed for biochemical, histological, immune-histochemical, and electron microscopic studies. Results: MSG caused thyroid follicles distortion and degeneration. Follicular cells possessed vacuolated cytoplasm, and pyknotic nuclei. Decreased colloids & inflammatory infiltrates were detected. By E/M, follicular cells showed dilated RER cisternae, degenerated mitochondria, & depleted microvilli. While basophils of pituitary gland appeared degenerated with vacuolated cytoplasm. By E/M, thyrotrophes showed dilated RER cisternae, dilated Golgi apparatus, and degenerated mitochondria. There was a highly significant decrease (P value˂0.001) in the mean serum level of T3, T4 and TSH in Group II) as compared with group I. While group III showed a highly significant increase (P value˂0.001) in the same parameter as compared with group II. The administration of PPE induced improvements of these changes.Conclusion: This work demonstrated that pomegranate peel extract improves the histological & biochemical changes induced by MSG on the pituitary thyroid axis.Key words: monosodium glutamate, pituitary thyroid axis, pomegranate peel extract.

Effect of Energy Drink on the Pancreas of Adult Male Albino Rat and the Possible Protective Role of Avocado Oil. Histological and Immunohistochemical Study

Introduction: Energy drinks are widely used among youth as boosters to increase their concentration. Many Health disorders associated with energy drinks have been reported. Avocado oil is a natural oil extracted from avocado fruit and possess an antioxidant and anti-inflammatory effects. Aim of the work: This work was conducted to investigate the effect of energy drink on the histological structure of the pancreas of adult male albino rat and to evaluate the possible protective role of avocado oil.Materials and Methods: Forty adult male albino rats were divided equally into 4 groups; control group, avocado oil-treated group (0.4 mg/100 gm body weight), energy drink-treated group (1.5 mg/100 gm body weight) and both avocado oil and energy drink treated group. All animals were orally administrated once daily for 4 weeks. Pancreatic specimens were processed for light and electron microscopy. Immunohistochemical study was performed using insulin and caspase-3 antibodies. Results: Energy drink-treated group revealed focal disturbed architecture of the exocrine and endocrine parts of pancreas. The exocrine pancreas appeared with cytoplasmic vacuolations and deeply stained nuclei of the acinar cells in addition to congested blood vessels. Ultrastructural examination revealed both acinar and islet cells with cytoplasmic vacuolations, swollen mitochondria with disrupted cristae, dilated rough endoplasmic reticulum in addition to shrunken condensed nuclei with dilated perinuclear cisternae. The immunohistochemical study showed a significant decrease in the insulin immunoreaction of β cells associated with a significant increase in caspase-3 immunoreaction in acinar and islet cells. In contrast, minimal changes were observed in rats treated concomitantly with avocado oil and energy drink with a non-significant change in the immunohistochemical reaction. Conclusion: Energy drink induced structural changes in the pancreas of rats and the concomitant intake of avocado oil could to some extent ameliorate such changes

Histological study of the possible protective action of platelet rich plasma on the injurious effect induced by aluminum chloride on albino male rat cerebellar cortex

Background: Aluminum compounds are commonly used in many human activities. They are used in pesticides, detergents, cosmetics, pharmaceuticals, and food additives. Many studies revealed a link between human exposure to aluminum and many neurodegenerative conditions in which there is platelet dysfunction.Aim of the study: The aim of this research was to study the potential protective effect of platelet rich plasma on the deleterious histological effect of aluminum chloride on male rat cerebellar cortex.Materials and methods: In this study, three equal groups of adult male albino rats were used; each consisted of 10 rats: group I a and I b (control group), group II (daily intraperitoneal injection of aluminum chloride dissolved in saline for 60 consecutive days in a dose of 10mg/kg) and group III were given PRP (subcutaneous injection in a dose of 0.5 ml/kg twice weekly) in addition to aluminum chloride in the same previous dose and duration. Cerebellum sections were processed for hematoxylin and eosin staining, GFAB staining and electron microscopic examination.Results: Aluminum chloride administration resulted in many histological alterations. Glial fibrillary acidic protein-positive cells were significantly present in the cerebellum of aluminum chloride-treated animals relative to control animals. Purkinje cells with darkly stained ill-defined nuclei with dark vacuolated cytoplasm with dilated RER cisterns, swollen mitochondria with destroyed crista were observed in an ultrastructural study for the cerebellar cortex of the aluminum chloride-treated group. It also showed degenerated granule cells with the disappearance of the nuclear membrane and vacuolation of the cytoplasm. Some of the myelinated nerve fibers showed degenerative changes. Concomitant administration of PRP decreased these effects.Conclusion: PRP partially minimized the severity of aluminum chloride-induced cerebellar cortex injurious histological effects in male albino rats. Further research is needed to extend these findings to clinical practice.

Impact of constant light exposure during pregnancy on skin of neonatal New Zealand rabbits: structural and ultrastructural study.

Continuous industrial productivity and modern societies have resulted in excess artificial light. The altered circadian rhythm causes many diseases. During intrauterine life, the mother's maternal melatonin rhythm has a major role in influencing organ development. The aim of this study was to investigate the effect of maternal exposure to constant light on the structure and ultrastructure of neonatal skin. Twenty pregnant New Zealand rabbits were divided into two groups (n=10 each): control group (12-h light/dark) and constant light group (24-h light). Plasma maternal melatonin and corticosterone during pregnancy were determined. At the end of the experiment, the dorsal skin of the neonates of both groups was collected and prepared for histological, morphometric, and transmission electron microscopic study. Histological and morphometric results of skin of neonates from the constant light group revealed statistically significantly reduced epidermal thickness, decreased number of hair follicle, increased surface area of collagen, and decreased proliferating cell nuclear antigen (PCNA) positive cells. Ultrastructural examination showed wide intercellular spaces and disrupted desmosomal junctions in the epidermis. Earlier stages of hair follicles were also observed with indented shrunken nuclei, vacuolization, and swollen mitochondria. Dermal fibroblasts with dilated cisternae of rough endoplasmic reticulum containing electron-dense material were detected. Maternal melatonin was significantly reduced in the constant light group while maternal corticosterone showed no significant difference between groups. Therefore, normal maternal circadian rhythm is a key factor for the integrity of neonatal skin structure.

Modulatory effect of bone marrow-derived stem cells against Tamoxifen induced liver injury in albino rats: A Histological, Immunohistochemical and Biochemical study

Tamoxifen (TAM) is commonly used for breast cancer treatment. It is considered non-steroidal antiestrogen agent. Toxic hepatitis, significant hepatic steatosis, and cirrhosis have been reported to be caused by TAM. Stem cells (BM-MSCs) are used for regenerative medicine and therapy. Aim of the work: to know how BMMSCs can improve tamoxifen induced damage in the liver of female albino rats. Materials & Methods: Ten female albino rats of average 200 gm were served as a donor for stem cells. Thirty female adult albino rats of 200-240 gm were randomly divided to three equal groups (10 animals each). Group I (Control group): provided ordinary diet. Group II (tamoxifen treated group): rats were orally given TAM with a dosage 20 mg per kg body weight daily for nine weeks consecutively. Group III (TAM & BM-MSCs group): animals were given TAM with a dosage 20 mg per kg body weight daily for nine weeks consecutively as in group II then received single intraperitoneal injection of 2×106 BM-MSCs suspended in PBS per rat then scarified after another four weeks. Biochemical studies were done, and liver sections were processed for histological, immunohistochemical and electron microscopic studies. Results: Sections of the animals’ liver taking TAM showed disturbance of the normal structure, showing cytoplasmic vacuolation, leukocytic infiltration, pyknotic nuclei, dilated cisternae of rough endoplasmic and dilated blood sinusoids also excessive collagen fibers, as well as increased expression of caspase 3. The levels of liver enzymes((AST), (ALT) and (ALP)) were elevated, On the other hand, animals’ liver sections treated with TAM followed by BM-MSCs showed improvement of the disturbed structure. In conclusion, BM-MSCs revealed ameliorative effect against TAM toxicity in rats.