Dilated Endoplasmic Reticulum Research Articles

Evaluation of tolerance to λ-cyhalothrin and response of detoxification enzymes in silkworms reared on artificial diet

A representative silkworm rearing mode of Ⅰ-Ⅲ instars reared on artificial diet and Ⅳ-Ⅴ instars reared on fresh mulberry leaves has been recognized in some sericultural areas of China. Under this rearing mode, silkworms are prone to be poisoned by pesticide residues on mulberry leaves at the Ⅳ and Ⅴ instar stages. As one of the most widely applied insecticides, λ-cyhalothrin was used to study the insecticide tolerance of silkworm reared on artificial diet (referred as the AD group). Our results showed that the newly ecdysized Ⅳ instar larvae in the AD group were less tolerant to λ-cyhalothrin compared to the mulberry leaves reared group (referred as the ML group). After continuous exposure to trace λ-cyhalothrin, the weight gain and the survival rate of silkworms were significantly lower in the AD group than those in the ML group, even though compensatory growth was observed in the control of the AD group. Histopathology and ultrastructure of fat body showed that λ-cyhalothrin induced more severe cell injuries in the AD group, such as shrunken nucleus, dilatation of endoplasmic reticulum, and mitochondrial swelling. The transcription levels of detoxification related genes (CYP4M5, CYP6AB4, CarE2, CarE5, GSTe1 and GSTe3) and the enzyme activities of P450s, CarEs and GSTs were inducible by trace λ-cyhalothrin in a time-specific manner, and the data showed that the response of P450 enzyme activity was retarded in the AD group, indicating a potential reason for a higher sensitivity to λ-cyhalothrin. Our results provided a new clue for the study of the relationship between feed nutrition and detoxification ability, and also provided an important reference for the development of modern silkworm rearing mode.

Ameliorating effect of selenium nanoparticles on cyclophosphamide-induced hippocampal neurotoxicity in male rats: light, electron microscopic and immunohistochemical study.

Cyclophosphamide (CPH) is a widely used chemotherapeutic drug that can affect the hippocampal neurocytes with a subsequent effect on memory and cognitive functions. Nanomedicine has the potential to overcome the current chemotherapeutic side effects, because of the unique nanoscale size and distinctive bioeffects of nanomaterials. So, the present study aims to investigate the potential ameliorative effect of the biologically synthesized nano-selenium (nano-Se) on CPH induced hippocampal neurotoxicity. Twenty four rats were randomly classified into four groups of 6 rats each: control group, nano-Se group (dose of 0.5 mg biological nano-Se/kg daily via oral gavage), CPH group (dose of 20 mg CPH/kg daily intraperitoneally), and CPH plus nano-Se group. After 4 weeks, the rats were sacrificed and the hippocampus was excised and processed. Sections were stained with haematoxylin and eosin stain and immunohistochemically stained for caspase-3 (apoptosis marker) and glial fibrillary acidic protein (astrocytic activity marker) (GFAP). Morphometric analysis and transmission electron microscopic (TEM) examination were also done. Control and nano-Se groups revealed no structural changes. By light microscopy, CPH group showed degeneration and necrosis of hippocampal neurocytes, significantly reduced thickness of the neurocyte cell layers, increased expression of GFAP and caspase-3 immunostains and significantly elevated apoptotic index. Moreover, neurocytes damage, mitochondrial cristeolysis, mild dilation of rough endoplasmic reticulum, and disrupted neurolemmal sheaths of nerve fibres were also demonstrated by TEM. Nano-Se cotreatment in the fourth group reversed all the aforementioned deleterious changes that induced by CPH in the hippocampal neurocytes. Treatment with CPH caused damage to hippocampal neurocytes that can be reversed by biological nano-Se co-treatment.

Loss of an ER component, membralin, in astrocytes triggers neuroinflammation.

During the progression of Alzheimer's disease (AD), and other neurodegenerative disorders, glial cells including astrocytes and microglia undergo morphological and physiological changes indicative of reactivity. However, whether glial activation is a non-consequential event from pathological stress or alternatively, whether glial activation can contribute to disease pathogenesis is unclear. Endoplasmic reticulum (ER) is the primary cellular organelle for the synthesis of transmembrane and secreted proteins. Previously, we identified an ER protein, membralin, as a novel component of the ER-associated degradation (ERAD) complex. Loss of membralin impaired homeostatic turnover of a key g-secretase complex subunit, nicastrin, resulting in increased g-secretase complex formation and activity, and aggravated AD pathology in TgCRND8 mice. Further, conditional deletion of membralin in astrocytes decreased expression of the excitatory amino acid transporter 2 (EAAT2) and induced excitotoxicity. Using adeno-associated viral (AAV) vectors to elevate membralin levels in the SOD1G93A ALS mouse model can significantly extend the lifespan of these animals. We use RNAseq, electron microscope, primary astrocyte culture, animal models in the study. Analysis of astroglial ER morphology using electron microscope shows that lose of membralin leads to dilated ER, which indicated elevation of ER stress. Transcriptomic analysis of motor cortex from astrocyte-specific membralin knockout animals reveals induction of reactive astrocyte profiles and disease-associated microglia (DAM) signatures. Further, membralin upregulation in astrocytes can alleviate neurotoxic (A1) astrocyte induction. Together, these results indicate that membralin is a critical neuroinflammation regulator. Modulation of astrocyte membralin levels may be promising as a neuroinflammatory suppressor in AD.

Protective effects of Rosemary extract and/or Fluoxetine on Monosodium Glutamate-induced hippocampal neurotoxicity in rat

The use of Monosodium Glutamate (MSG) as a food flavor enhancer is increasing worldwide despite its neurotoxic effects. Fluoxetine (FLX) and Rosemary extract (RE) are known to have beneficial neuroprotective properties. Rats were divided into five groups: control group; MSG group, rats received 2 g/kg/day intraperitoneal (i.p.) injections of MSG for seven days; RE/MSG group, rats received 50 mg/kg/day of oral RE for 28 days starting prior to MSG; FLX/MSG group, rats received 10 mg/kg/day of oral FLX for 28 days beginning before MSG; and RE/FLX/MSG group, received combined treatments as mentioned above. Rats underwent the Barnes maze test, in addition to histopathological, immunohistochemical, morphometric and ultrastructural evaluations for their hippocampi. MSG increased the number of errors and escaped latency in the Barnes maze test that was significantly minimized in the three treatment groups. The MSG group exhibited pyramidal cell (PC) degeneration, shrunken glial cells and massive vascular dilatation that were improved with RE and/or FLX treatment. The number of glial fibrillary acidic protein (GFAP)-immunopositive cells were increased, and the number of PCs was decreased in the MSG group, while these values were significantly reversed with the three treatment groups with the most significant improvement at RE/FLX/MSG one. Ultrastructurally, PCs were shrunken with degenerated nuclei, dilated endoplasmic reticulum, swollen mitochondria, and vacuolations in the MSG group that were improved with RE and/or FLX. In conclusion, the combined RE and FLX treatment can ameliorate the toxic effect of MSG on rat hippocampus probably through its antioxidant and anti-inflammatory effects.

Possible Disrupting Effect of Different Doses of Butylated hydroxyanisole (BHA) on Thyroid Follicular Cells of Adult Male Albino Rats: Anatomical, Histological & Biochemical Study

Background:Butylated hydroxyanisole (BHA) is a synthetic antioxidant which is absorbed mainly through gastrointestinal tract following oral administration. It was introduced to the food industry as a safe preservative and to the pharmaceutical industry in cosmetics manufacturing and as an additive to fat-soluble vitamins to prevent rancidity. The US Food and Drug Administration (FDA) approves BHA as a safe food additive in a maximum oral dose of 150 mg/ kg, meanwhile it is a potential allergen. A new question has been raised if BHA is a potential human carcinogen? another potential hazard is also a recent concern as the compound has been linked with some endocrine disruption. The endocrine effect of BHA is associated with sex hormones, a little has been known about the exact effect neither on them nor on other hormones.Aim of the work:This work aimed to assess the possible disrupting effect of different doses of BHA on thyroid follicular cells.Materials and methods:This study was conducted on 30 adult male albino rats. 12 as control and 18 divided equally to 3 groups and received 10, 100 and 500 mg/kg body weight of BHA dissolved in corn oil orally for 42 days. All animals were examined for body and thyroid weight, thyroid hormones and oxidative markers levels. Semithin and ultrastructural examination of thyroid follicles were done for all. All data were statistically analyzed.Results:All data revealed dose defendant disrupting effect of BHA on thyroid and follicular cells. Variable follicular shapes and multiple layers of vacuolated cells and dilated endoplasmic reticulum. Histological findings were confirmed by both anatomical and biochemical examinations.conclusion:BHA has a possible thyroid disrupting effect in long term intake. This effect is dose dependent and significantly recognized in moderate and high doses intake.

COPB2 loss of function causes a coatopathy with osteoporosis and developmental delay

Coatomer complexes function in the sorting and trafficking of proteins between subcellular organelles. Pathogenic variants in coatomer subunits or associated factors have been reported in multi-systemic disorders, i.e., coatopathies, that can affect the skeletal and central nervous systems. We have identified loss-of-function variants in COPB2, a component of the coatomer complex I (COPI), in individuals presenting with osteoporosis, fractures, and developmental delay of variable severity. Electron microscopy of COPB2-deficient subjects' fibroblasts showed dilated endoplasmic reticulum (ER) with granular material, prominent rough ER, and vacuoles, consistent with an intracellular trafficking defect. We studied the effect of COPB2 deficiency on collagen trafficking because of the critical role of collagen secretion in bone biology. COPB2 siRNA-treated fibroblasts showed delayed collagen secretion with retention of type I collagen in the ER and Golgi and altered distribution of Golgi markers. copb2-null zebrafish embryos showed retention of type II collagen, disorganization of the ER and Golgi, and early larval lethality. Copb2+/- mice exhibited low bone mass, and consistent with the findings in human cells and zebrafish, studies in Copb2+/- mouse fibroblasts suggest ER stress and a Golgi defect. Interestingly, ascorbic acid treatment partially rescued the zebrafish developmental phenotype and the cellular phenotype in Copb2+/- mouse fibroblasts. This work identifies a form of coatopathy due to COPB2 haploinsufficiency, explores a potential therapeutic approach for this disorder, and highlights the role of the COPI complex as a regulator of skeletal homeostasis.

Effect of long-term exposure to acrylamide on endoplasmic reticulum stress and autophagy in rat cerebellum

Acrylamide (ACR) is a widely used chemical compound that has neurotoxicity in human, but whether ACR could impair the cerebellum and the related mechanism were still unknown. This study aimed to observe the changes in behavioral performance and cerebellar morphology caused by chronic ACR exposure, and to evaluate its influence on apoptosis, endoplasmic reticulum stress (ERS) and autophagy. Rats were treated with 0, 0.5 and 5 mg/kg ACR by drinking water for 12 months. Results showed that 5 mg/kg ACR treatment damaged the gait, balance ability, hindlimb muscle strength and motor coordination ability of rats. The results of hematoxylin and eosin and Nissl staining indicated that ACR impaired the structures of all three layers of the cerebellum, especially the Purkinje cell layer, showing abnormal morphology with nucleus condensation and pyknosis. Accumulation of autophagosomes, dilated endoplasmic reticulum and swollen mitochondria were observed in neurons under transmission electron microscopy. The enhanced apoptotic rates and the increased Bax expression indicated the elevated level of apoptosis. The results of Western blot showed that ACR treatment elevated protein levels of Beclin1, LC3-II/LC3-I, p-PERK/t-PERK, ATF4 and CHOP, indicating the initiation of autophagy, the activation of PERK pathway in ERS. This work helps to demonstrate the ACR neurotoxicity on cerebellum under chronic treatment and its underlying mechanism.

Metal-coordinated nanomedicine for combined tumor therapy by inducing paraptosis and apoptosis

Apoptosis resistance of tumor cells often results in chemoresistance and treatment failure in clinic. In this work, a Cu2+-coordinated morusin/doxorubicin biological organizer (designated as COMBO) is designed to combat cellular resistance to apoptosis for combined tumor therapy. By virtue of the coordination and π-π stacking effects, the self-assembled COMBO possesses nanometer particle size, narrow and homogenous graininess distribution as well as a good dispersion stability. Moreover, COMBO could be disassembled by glutathione (GSH) with an effective drug release and fluorescence recovery. Morusin-mediated paraptosis could induce extensive vacuolization through the dilation of endoplasmic reticulum (ER) and mitochondria, leading to non-apoptotic programmed cell death (PCD) regardless of the cellular resistance to apoptosis. Furthermore, the released doxorubicin prefers to locate in cell nucleus to cause cell apoptosis for combined chemotherapy. By the joint action of paraptosis and apoptosis, COMBO exhibits a great superiority over monotherapy in tumor inhibition with a low system toxicity. This study may open a window in the development of self-delivery nanomedicine for overcoming apoptosis resistance in tumor therapy.

Telocytes (interstitial cajal like cells) of the fallopian tubes in acute and chronic salpingitis

Telocytes (TCs) are a recently described population of cells. Their histogenesis, phytophysiology, function and role in the development of pathological conditions are discussed by many researchers. In particular, the pathophysiological role of TCs in the fallopian tubes affected by inflammation remains unexplored. Objective: to study ultrastructural changes in the TC and their diagnostic significance in the distal fallopian tubes in acute and chronic salpingitis. Histological, immunohistochemical (IHC) and electron microscopic (EM) examination of fragments of the distal fallopian tubes of 10 women (age 37.8 ± 7.1 years) with acute and chronic inflammation, as well as without disease were carried out. IHC study was carried out with antibodies characteristic of stem cells (CD34, CD117) and Cajal cells (CD117, DOG1). We believe that immunophenotyping with well-known markers is not specific and is not suitable for the identification of TCs in the fallopian tubes. An EM study revealed multiple destructive changes in the TC. In acute salpingitis - loss of organelles, cytoplasmic vacuolization, dilatation of the granular endoplasmic reticulum, loss of intercellular contacts, shortening of telopodia, local loss of plasma membrane integrity. In chronic salpingitis, there is an increase in destructive changes and a further decrease in the number of TCs. Changes in TC and a decrease in their number alter the three-dimensional organization of the extracellular matrix in the stromal compartment of the fallopian tube, weaken intercellular signaling and contractility of the fallopian tube, which contributes to impaired organ motility.

Regulation of Cyclic Nucleotide Phosphodiesterase 1 by Endoplasmic Reticulum Stress Contributes to Homocysteine‐induced Cardiac Hypertrophy

Objectives Mechanistic understanding of the role of homocysteine in the development of cardiac hypertrophy is inadequate at present. This study aimed to elucidate the association of cyclic nucleotide phosphodiesterase 1 (PDE1) and endoplasmic reticulum (ER) stress with cardiomyocyte hypertrophy induced by homocysteine. Methods H9c2 cardiomyocytes were allocated to control group and homocysteine-treated group in the presence or absence of ER stress inhibitor tauroursodeoxycholic acid (TUDCA) or PDE1 inhibitor Lu AF58027 for 72h. In another set of experiment, H9c2 cells were transfected with specific siRNA targeting PDE1 subtypes prior to homocysteine exposure. The cell surface area was measured by phase contrast microscope and image analysis software and ultrastructural changes of the cells were observed under transmission electron microscopy. Quantitative polymerase chain reaction and western blot were employed to detect the mRNA and protein expression of the hypertrophic markers beta-smooth muscle myosin heavy chain (β-MHC) and atrial natriuretic peptide (ANP), PDE1 subtypes (PDE1A, 1B, and 1C), and ER stress molecules (GRP78, phosphor-PERK, PERK, phosphor-IRE1, IRE1, and ATF6). The intracellular levels of cyclic guanosine monophosphate (cGMP) and cyclic adenosine monophosphate (cAMP) were measured by enzyme linked immunosorbent assay. Results Homocysteine exposure resulted in an enlargement in cell size and an increase in β-MHC and ANP expression. Homocysteine induced ER stress in H9c2 cells, shown by increased expression of GRP78 and enhanced phosphorylation of PERK and IRE1, as well as dilatation and degranulation of rough ER. The expression of PDE1A and PDE1C was significantly upregulated by homocysteine. Inhibition of ER stress or PDE1 suppressed homocysteine-induced upregulation of β-MHC and ANP and alleviated cell enlargement. In addition, the protein expression of PDE1A and PDE1C in homocysteine-exposed H9c2 cells was downregulated by the ER stress inhibitor TUDCA. Silencing of PDE1C significantly inhibited homocysteine-induced cell hypertrophy whereas knockdown of PDE1A showed minor effect. Both cAMP and cGMP levels were lowered in H9c2 cells subjected to homocysteine exposure, while only cAMP content could be restored by Lu AF58027 and TUDCA. Conclusions ER stress mediates homocysteine-induced cardiomyocyte hypertrophy via upregulating PDE1 expression. Cyclic nucleotide, in particular cAMP, is the downstream mediator of the ER stress-PDE1 signaling axis in homocysteine-induced hypertrophy.

Ca2+ overload- and ROS-associated mitochondrial dysfunction contributes to \u03b4-tocotrienol-mediated paraptosis in melanoma cells

Melanoma is an aggressive tumor with still poor therapy outcomes. δ-tocotrienol (δ-TT) is a vitamin E derivative displaying potent anti-cancer properties. Previously, we demonstrated that δ-TT triggers apoptosis in human melanoma cells. Here, we investigated whether it might also activate paraptosis, a non-canonical programmed cell death. In accordance with the main paraptotic features, δ-TT was shown to promote cytoplasmic vacuolization, associated with endoplasmic reticulum/mitochondrial dilation and protein synthesis, as well as MAPK activation in A375 and BLM cell lines. Moreover, treated cells exhibited a significant reduced expression of OXPHOS complex I and a marked decrease in oxygen consumption and mitochondrial membrane potential, culminating in decreased ATP synthesis and AMPK phosphorylation. This mitochondrial dysfunction resulted in ROS overproduction, found to be responsible for paraptosis induction. Additionally, δ-TT caused Ca2+ homeostasis disruption, with endoplasmic reticulum-derived ions accumulating in mitochondria and activating the paraptotic signaling. Interestingly, by using both IP3R and VDAC inhibitors, a close cause-effect relationship between mitochondrial Ca2+ overload and ROS generation was evidenced. Collectively, these results provide novel insights into δ-TT anti-melanoma activity, highlighting its ability to induce mitochondrial dysfunction-mediated paraptosis.Graphic δ-tocotrienol induces paraptotic cell death in human melanoma cells, causing endoplasmic reticulum dilation and mitochondrial swelling. These alterations induce an impairment of mitochondrial function, ROS production and calcium overload.

BIMG-10. IDH1 MUTATIONS INDUCE ORGANELLE DEFECTS VIA DYSREGULATED PHOSPHOLIPIDS

BACKGROUNDMetabolic alterations of lipids have been identified as a hallmark of neoplasms, with the most prevalent being the balance between saturated fatty acid (SFA) and monosaturated fatty acid (MUFA). Stearoyl-CoA desaturase1 (SCD1), converting SFA to MUFA, is increased in many cancers, leading to worse prognosis. In glioma, the role of SCD1 remains unknown. Isocitrate dehydrogenase (IDH) mutations have been most commonly observed in glioma, but the involvement of mutant IDH in SCD1 expression also remains unknown.METHODSWe conducted metabolic analysis to examine the alteration of SCD1 expression in genetically engineered glioma cell lines and normal human astrocyte (NHA). Lipid metabolic analysis was conducted by using LC-MS, Raman Imaging Microscopy and SCD1 expression was examined by Western-blotting and RT-PCR method. Electron microscopy was employed for organelle structure and genetic knock-down of SCD1 gene was performed.RESULTHerein, we uncovered increased MUFA and their phospholipids in Endoplasmic Reticulum (ER), generated by IDH1 mutation, that were responsible for Golgi and ER dilation. RNA seq data from The Cancer Genome Atlas, showed that SCD1 expression was significantly higher in IDH mutant gliomas compared with wild-type, and high SCD1 expression was associated with longer survival. Inhibition of IDH1 mutation or SCD1 silencing restored ER and Golgi morphology, while D-2HG and oleic acid induced morphological defects in these organelles. Moreover, addition of oleic acid, which tilts the balance towards elevated levels of MUFA, produced IDH1 mutant-specific cellular apoptosis.CONCLUSIONCollectively, our results suggest that IDH1 mutant-induced SCD overexpression can rearrange the distribution of lipids in the organelles of glioma cells, providing a new insight on the link between lipids metabolism and organelle morphology in these cells, with potential and unique therapeutic implications. The results of the present study may also provide novel insights into the discovery of metabolic biomarkers for IDH mutant gliomas.

Ameliorating Effect of Platelet- rich plasma on Histological and Biochemical Changes Induced by Bisphenol-A in Prostate of Adult Albino Rats

The incidence of male reproductive disorders has been increased and may be linked to the environmental natural and synthetic chemicals. These chemicals are known as endocrine disrupting chemicals (EDCs); Bisphenol-A (BPA) is the most common one. Platelet rich plasma (PRP) is an autogenous and economical source of growth factors which nowadays used in the tissue repair. So, this study aimed to use the histological and immunehistochemical approaches for evaluating the possible therapeutic effect of PRP against BPA induced prostatic lesions in adult albino rats. Thirty five adult male albino rats were divided into 3 main groups. Group I served as control, Group II in which the rats were received 50 mg/kg BPA once daily for 4 weeks, Group III in which the rats were received BPA as group II then 0.5 ml of PRP subcutaneous injected twice weekly for 4 weeks. After 8 weeks, rats of all groups were subjected to the followings; biochemical analysis of serum testosterone, immunohistochemical studies for androgen and estrogen receptors (AR &ER), light and electron microscope examination. Serum testosterone levels showed a highly significant decrease in group II when compared to others. Histologically, BPA treated prostate showed abundant collagen fibers with congested blood vessels and inflammatory cellular infiltration while, their acini showed numerous folds with epithelial stratification. Ultrastructurally, the secretory cells had heterochromatic nuclei, dilated endoplasmic reticulum, apical few microvilli and polymorphic secretory granules with lysosomes. Nearly all acinar cells showed negative nuclear ARs while positive ERs detection in basal cells. After PRP administration, showed amelioration of many of these changes. We concluded that exposure to BPA at low dose induced deleterious effects on the histological structure of the prostate and disturbed reproductive hormones. PRP showed significant signs of restoration of these changes.

Quantitative proteomic analysis reveals novel insights into hepatic toxicity in mice exposed chronically to okadaic acid

Okadaic acid (OA) is a principal shellfish toxin and a potent promoter of hepatic tumors. However, the molecular mechanisms of OA-induced susceptibility to carcinogenesis are largely unknown. Here, we applied a quantitative proteomic approach, two-dimensional differential gel electrophoresis, to characterize the differentially expressed proteins of hepatic tissue in adult male mice exposed chronically to OA (0.2, 2, and 10 μg/kg body weight) for 120 days via daily intraperitoneal injection. Our results showed that liver morphology was damaged by OA, and dilation of rough endoplasmic reticulum, swelling and ridge disappearance of mitochondria in hepatocytes were observed in all OA-treated mice. Serum malondialdehyde content increased significantly with increasing OA dose while hepatic protein phosphatase activity decreased. Furthermore, apoptotic cells in the liver showed a significant dose-response relationship. Quantitative proteomic analysis revealed that 46 proteins altered remarkably in abundance in OA-treated mice, and these proteins were involved in macromolecular metabolism, molecular chaperone/stress response, apoptosis, and cytoskeleton. Bioinformatic analysis indicated that OA might induce apoptosis by activating extracellular signal-regulated kinase-1/2, c-Jun N-terminal kinase, and nuclear factor-kappa B pathways, as well as causing endoplasmic reticulum stress, thereby leading to liver injury. OA-induced hepatocyte apoptosis might be a key mechanism responsible for hepatotoxicity. We showed, for the first time, the chronic toxic effects of OA on mice at the mechanistic level.

Histomorphometric Evidence of Hepatic Recovery of Rats Fed Repeatedly Heated Palm Oil

ABSTRACTBackground: Repeated heating of vegetable oils especially palm oil that is commonly and commercially used in frying of food at homes and restaurants has been investigated for its risks on health status. It seemed to carry potential hazards on the liver tissue. Aim: To investigate the impact of eating foods fried with 5-times heated palm oil on liver of adult male rats for 8 weeks and the possibility of recovery of histomorphometric changes after stoppage of use of reheated palm oil for additional 4 weeks.Material and Methods: Thirty adult male albino rats were divided equally into 3 groups; control group (received normal balanced diet for 8 weeks), H5 group (received 5-times heated palm oil enriched diet for 8 weeks) and Recovery group (received five times heated palm oils for 8 weeks and then left on balanced diet for another 4 weeks). At the end of the experiment, animals were anesthetized with ether then sacrificed. Specimens taken from the liver were processed for histological evaluation. Morphometric studies were conducted and collected data undergone statistical analysis.Results: Histological examination showed alterations in liver tissue of H5 group including fat droplets, mononuclear cellular infiltration, dilatation of central vein and blood sinusoids, collagen deposition and degeneration and necrosis of hepatocytes. Ultrastructural examination showed lipid droplets, irregular mitochondria, cytoplasmic vacuolation, irregular nuclear envelop and dilatation of endoplasmic reticulum. Recovery group showed decreased lipid droplets, less dilatation and congestion of central vein and blood sinusoids, decreased inflammatory cells but few hepatocytes still show degeneration. Ultrastructure of hepatocytes of recovery group became more or less normal with restoration of cytoplasmic organelles. Morphometric results support the histological findings.Conclusion: Consumption of repeatedly heated palm oil is injurious to the liver tissue, and those injuries show possible recovery after stoppage of its consumption.

IDH1 mutations induce organelle defects via dysregulated phospholipids

Infiltrating gliomas are devastating and incurable tumors. Amongst all gliomas, those harboring a mutation in isocitrate dehydrogenase 1 mutation (IDH1mut) acquire a different tumor biology and clinical manifestation from those that are IDH1WT. Understanding the unique metabolic profile reprogrammed by IDH1 mutation has the potential to identify new molecular targets for glioma therapy. Herein, we uncover increased monounsaturated fatty acids (MUFA) and their phospholipids in endoplasmic reticulum (ER), generated by IDH1 mutation, that are responsible for Golgi and ER dilation. We demonstrate a direct link between the IDH1 mutation and this organelle morphology via D-2HG-induced stearyl-CoA desaturase (SCD) overexpression, the rate-limiting enzyme in MUFA biosynthesis. Inhibition of IDH1 mutation or SCD silencing restores ER and Golgi morphology, while D-2HG and oleic acid induces morphological defects in these organelles. Moreover, addition of oleic acid, which tilts the balance towards elevated levels of MUFA, produces IDH1mut-specific cellular apoptosis. Collectively, these results suggest that IDH1mut-induced SCD overexpression can rearrange the distribution of lipids in the organelles of glioma cells, providing new insight into the link between lipid metabolism and organelle morphology in these cells, with potential and unique therapeutic implications.

Glial activation and inflammation in the NTS in a rat model after exposure to diesel exhaust particles

Airway pollution can affect the central nervous system, but whether this causes glial activation and inflammation in the nucleus of solitary tract (NTS) remains unclear. We used a rat model with exposure to diesel exhaust particulate matter (DEP) at 200 μg/m3 (low exposure) and 1000 μg/m3 (high exposure) for 14 days. Activation of microglia and astrocytes in the NTS was assessed using Iba-1 and glial fibrillary acidic protein (GFAP) staining. The expression of neurotrophic factors including brain-derived neurotrophic factor (BDNF), glial-derived neurotrophic factor (GDNF), and nerve growth factor (NGF) in the NTS were evaluated by immunofluorescence. Changes in the intracellular structure of NTS neurons were observed via electron microscopy. Inflammatory cytokines and oxidant stress levels in the medulla were also measured. Exposure to DEP can cause NTS inflammation as well as airway inflammation, especially in the H-exposure group. We showed that the numbers of microglia and astrocytes in the NTS, as well as NGF expression in the NTS, were significantly higher in both exposure groups than in controls, but BDNF or GDNF expression was not detected. Exposure to DEP induced ultrastructural changes in NTS neurons as reflected by endoplasmic reticulum dilation, ribosomal loss, mitochondrial vacuolization, and a sparse myelin sheath. Medulla inflammation and an imbalance of oxidants and antioxidants also resulted from exposure to DEP. The H-exposure group showed an imbalance of oxidants and antioxidants with decreased levels of SOD and GSH and increased levels of MDA and ROS compared to the control group (both p < 0.01) in the medulla. Inflammatory cytokines (IL-1β, IL-6, and TNF-α) were also significantly increased in the H-exposure group. Fourteen days of exposure to DEP can affect the NTS neurons in rat. Glial activation and inflammation may play important roles in the response of the NTS to DEP.

Intracellular Ca2 + Imbalance Critically Contributes to Paraptosis.

Paraptosis is a type of programmed cell death that is characterized by dilation of the endoplasmic reticulum (ER) and/or mitochondria. Since paraptosis is morphologically and biochemically different from apoptosis, understanding its regulatory mechanisms may provide a novel therapeutic strategy in malignant cancer cells that have proven resistant to conventional pro-apoptotic treatments. Relatively little is known about the molecular basis of paraptosis, but perturbations of cellular proteostasis and ion homeostasis appear to critically contribute to the process. Ca2+ transport has been shown to be important in the paraptosis induced by several natural products, metal complexes, and co-treatment with proteasome inhibitors and certain Ca2+-modulating agents. In particular, the Ca2+-mediated communication between the ER and mitochondria plays a crucial role in paraptosis. Mitochondrial Ca2+ overload from the intracellular Ca2+-flux system located at the ER–mitochondrial axis can induce mitochondrial dilation during paraptosis, while the accumulation of misfolded proteins within the ER lumen is believed to exert an osmotic force and draw water from the cytoplasm to distend the ER lumen. In this process, Ca2+ release from the ER also critically contributes to aggravating ER stress and ER dilation. This review focuses on the role of Ca2+ transport in paraptosis by summarizing the recent findings related to the actions of Ca2+-modulating paraptosis-inducing agents and discussing the potential cancer therapeutic strategies that may effectively induce paraptosis via Ca2+ signaling.

Renal Alterations Induced by Chronic Exposure to Therapeutic Doses of Antihypercholestremic Atorvastatin.

Atorvastatin (ATOR) is widely used for the treatment and prevention of hypercholesterolemia and various diseases, such as cardiovascular complications, with little data about the histopathological and ultrastructural renal alterations that might be induced by this drug. The present study was undertaken to investigate the potential toxicity of therapeutic doses of atorvastatin on the microanatomy and ultrastructure of renal tissues from Wistar albino rats. Adult male Wistar albino rats received an oral daily dose of 5 mg/kg bodyweight for 90 consecutive days. Biopsies from both kidneys of each study rat were taken for histopathological and ultrastructural examination. ATOR-treated rats exhibited glomerular, tubular, and interstitial histological alterations, including degeneration, necrosis, hyaline droplets, edema, cortical hemorrhages, mesangial hypercellularity, and blood capillary dilation and congestion. In addition, ATOR exposure increased the activity of glucose-6-phosphate dehydrogenase and alkaline phosphatase with a concurrent reduction in proteins and neutral mucosubstances content of the glomeruli and renal cells. Moreover, ATOR-treated animals demonstrated glomerular ultrastructural alterations, consisting mainly of capillary tuft dilatation, glomerular basement membrane thickening, and mesangial cell proliferation. The renal cells of the proximal tubules demonstrated damaged mitochondria, degenerative cellular changes, endoplasmic reticulum dilatation, lysosomal and autophagosome activation, nuclear alteration, myelin figure formation, and microvilli disorganization. The findings of the present work may indicate that ATOR can induce renal histological, histochemical, and ultrastructural alterations that may affect kidney and other vital organ functions.

Effect of Sofosbuvir (Sovaldi) on the fundic mucosa of adult male albino rats and the possible protective role of Fucoidan: Histological, histochemical, and immunohistochemical study

Background: Sofosbuvir (SOF) is a hopeful remedy for chronic hepatitis C virus infection. However, it records some unfavorable influences on gastrointestinal tract. Fucoidan is a sulfated polysaccharide having cytoprotective effect besides anti-inflammatory and antioxidant properties.Aim: This work has been focused on the possible effect of sofosbuvir on the fundic mucosa and to assess the probable role of protection of fucoidan.Materials and methods: we divided 40 adult male albino rats equally into four groups and were received drugs as a single daily dose for five weeks. Control group (I): were received distilled water and normal saline. Fucoidan group (II): were received 80 mg/kg. Sofosbuvir group (III): were received 4 mg/kg. Group IV: were received Fucoidan and Sofosbuvir with the same doses as pervious groups. All rats were sacrificed one day following the last dose. Stomach specimens were obtained and prepared for light and electron microscopic examination.Results: Sofosbuvir-treated rats showed dilated fundic pits and cystic dilatation of fundic glands with flattening of their epithelial lining. Vacuolation of cytoplasm and shrunken pyknotic nuclei were detected in parietal cells. Dilatation of rough endoplasmic reticulum and loss of secretory granules were detected in the chief cells. Infiltration of inflammatory cells with congested blood vessels and extravasated RBCs were detected in lamina propria. Statistically, there was a highly significant rise in the area% of collagen fibers while area% of Periodic Acid Schieff -Alcian Blue stain (PAS–AB) reaction showed a highly significant reduction comparing with the control. The immunohistochemical results revealed marked increase in proliferating cell nuclear antigen (PCNA) expressions. The administration of Fucoidan with Sofosbuvir minimized these changes.Conclusion: Sofosbuvir has been proved to induce histological changes in the fundic mucosa and these changes can be attenuated by Fucoidan when given in concomitant with it.